U.S. patent application number 15/890552 was filed with the patent office on 2018-11-29 for delivery of large molecular weight biologically active substances.
This patent application is currently assigned to PhotoKinetix Holdings, Inc.. The applicant listed for this patent is PhotoKinetix Holdings, Inc.. Invention is credited to Edward R. Kraft, Gabriela Kulp.
Application Number | 20180339166 15/890552 |
Document ID | / |
Family ID | 44626990 |
Filed Date | 2018-11-29 |
United States Patent
Application |
20180339166 |
Kind Code |
A1 |
Kraft; Edward R. ; et
al. |
November 29, 2018 |
DELIVERY OF LARGE MOLECULAR WEIGHT BIOLOGICALLY ACTIVE
SUBSTANCES
Abstract
The invention relates generally to intradermal, transdermal,
and/or transmembrane delivery of biologically active substances in
the epidermis and/or through the skin, sub-dermal tissues, blood
vessels and cellular membranes without causing damage to the
cellular surface, tissue or membrane. The biologically active
substances may have a molecular weight no less than about 5.8 kDa
to about 2,500 kDa, such as Hyaluronic Acid (HA). The biologically
active substances may be deposited in a dermal patch containing a
red algae polysaccharide-based matrix, wherein the red algae
polysaccharide is an extract of Chondrus crispus at 2% by weight of
the dermal patch. The invention provides systems and methods for
enhanced intradermal, transdermal, and/or transmembrane delivery of
such biologically active substances using pulsed incoherent light.
The invention further provides a device for the application of the
pulsed incoherent light to cellular surfaces and membranes using
those compositions and methods.
Inventors: |
Kraft; Edward R.;
(Galveston, TX) ; Kulp; Gabriela; (Santa Fe,
TX) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
PhotoKinetix Holdings, Inc. |
Galveston |
TX |
US |
|
|
Assignee: |
PhotoKinetix Holdings, Inc.
Galveston
TX
|
Family ID: |
44626990 |
Appl. No.: |
15/890552 |
Filed: |
February 7, 2018 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
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15292703 |
Oct 13, 2016 |
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15890552 |
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14008047 |
Jan 6, 2014 |
9474911 |
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PCT/EP11/58293 |
May 20, 2011 |
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15292703 |
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61468755 |
Mar 29, 2011 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61N 5/062 20130101;
A61K 38/30 20130101; C12Y 304/24069 20130101; A61K 38/27 20130101;
A61K 8/9789 20170801; A61N 5/0613 20130101; A61K 9/7046 20130101;
A61K 8/9717 20170801; A61K 8/645 20130101; A61K 9/0014 20130101;
A61K 45/06 20130101; A61N 2005/065 20130101; A61K 41/17 20200101;
A61N 2005/0659 20130101; A61K 8/345 20130101; A61Q 19/00 20130101;
A61K 38/4893 20130101; A61K 47/10 20130101; A61N 2005/0652
20130101; A61N 2005/0662 20130101; A61K 9/7023 20130101; A61N
2005/0661 20130101; A61K 47/46 20130101; A61K 31/728 20130101; A61K
8/0208 20130101 |
International
Class: |
A61N 5/06 20060101
A61N005/06; A61K 8/9717 20170101 A61K008/9717; A61Q 19/00 20060101
A61Q019/00; A61K 8/02 20060101 A61K008/02; A61K 8/34 20060101
A61K008/34; A61K 8/64 20060101 A61K008/64; A61K 9/00 20060101
A61K009/00; A61K 47/46 20060101 A61K047/46; A61K 47/10 20170101
A61K047/10; A61K 45/06 20060101 A61K045/06; A61K 41/00 20060101
A61K041/00; A61K 38/48 20060101 A61K038/48; A61K 38/30 20060101
A61K038/30; A61K 38/27 20060101 A61K038/27; A61K 31/728 20060101
A61K031/728; A61K 9/70 20060101 A61K009/70 |
Claims
1. A device for photokinetic intradermal and/or transdermal
delivery of a biologically active substance to a subject, said
device comprising: (1) a generator that provides an oscillating
electrical pulse; (2) at least one light emitting diode that
receives the oscillating electrical pulse and responds by providing
an incoherent light; and, (3) a donor cell that holds a formulation
comprising the biologically active substance, wherein the donor
cell is positioned to receive the incoherent light.
2-54. (canceled)
55. A method for photokinetic intradermal and/or trans dermal
delivery of a biologically active substance having a molecular
weight of no less than 5800 Da to a subject, the method comprising:
(1) preparing a formulation comprising the biologically active
substance; (2) applying said formulation to a cellular surface on
the skin of the subject; (3) illuminating said formulation on said
cellular surface with a pulsed incoherent light having a selected
wavelength, pulse rate and duty cycle; and, (4) allowing said
biologically active substance in said formulation to permeate said
cellular surface, thereby effecting photokinetic intradermal and/or
transdermal delivery of the biologically active substance.
56-61. (canceled)
62. The method of claim 55, wherein the formulation comprises a
gelling agent.
63. (canceled)
64. The method of claim 55, wherein the fornlulation further
comprises a photocatalytic agent.
65-72. (canceled)
73. The method of claim 55, wherein said biologically active
substance comprises sodium hyaluronate (e.g., one with a M.W. of at
least about 1000 kDa, 1600 kDa, 2200 kDa, 2500 kDa, 3000 kDa, 3500
kDa, 4000 kDa, 4500 kDa, 5000 kDa).
74. The method of claim 55, wherein said biologically active
substance is a cosmetic agent, or a therapeutic agent.
75. The method of claim 55, wherein said biologically active
substance comprises: (1) a peptide selected from the group
consisting of Gly-Tyr, Val-TyrVal, Tyr-Gly-Gly-Phe-Met (SEQ ID NO:
1), Tyr-Gly-Gly-Phe-Leu (SEQ ID NO: 2), and
Asp-Arg-Val-Tyr-Ile-His-Pro-Phe (SEQ ID NO: 3); (2) a honnone
selected from the group consisting of methionine enkephalin
acetate, leucine enkephalin, angiotensin II acetate, f)-estradiol,
methyl testosterone, and progesterone; or, (3) a protein selected
from the group consisting of enzymes, nonenzymes, antibodies, and
glycoproteins.
76. (canceled)
77. The method of claim 55, wherein said pulsed incoherent light is
selected from the group consisting of fluorescent, ultraviolet,
visible, near infrared, LED (light emitting diode), and halogen
light.
78-83. (canceled)
84. The method of claim 55, wherein said pulse rate is between
about 1.7 cycles per second (cps) and about 120 cps, or about 1.7
cps and about 80 cps.
85. (canceled)
86. The method of claim 55, wherein said duty cycle is between
about 50% and about 75%.
87. The method of claim 55, further comprising adjusting the
transdermal flux rate of the biologically active substance by
modulating the light energy.
88-110. (canceled)
Description
BACKGROUND OF THE INVENTION
[0001] This invention relates to photokinetic delivery of
biologically active substances from an outer mammalian skin
surface. Depending on specific needs, the biological substance can
be delivered intradermally (e.g., substantially locally to the
epidermis level by, for example, predicting the intradermal drug
deposition over time and modulating that deposition through turning
off the light and thus the photokinetic process), for example, in
cosmetic uses in the skin, or be delivered transdermally (e.g., to
an underlying tissue or blood vessel), for example, in
pharmaceutical uses. The biological substance can also be delivered
from an extracellular environment to intracellular environment
(transmembrane).
[0002] More particularly, the invention provides compositions for
enhanced intradermal, transdermal, or transmembrane delivery of
biologically active substances using pulsed incoherent light. In
addition, the invention provides methods and devices for
application of pulsed incoherent light to an area of mammalian skin
or membrane for safe and efficient intradermal, transdermal, or
transmembrane delivery of biologically active substances into or
through the skin surface or cellular membrane.
[0003] For example, in the pharmaceutical field, therapeutic agents
or biologically active substances can be administered to vital
tissues and organs in a mammal by a plethora of delivery routes
including, for example, oral, nasal, aural, anal, dermal, ocular,
pulmonary, intravenous, intramuscular, intra-arterial,
intraperitoneal, mucosal, sublingual, subcutaneous, and
intracranial routes. In the last decade, transdermal delivery of
biologically active substances has gained momentum due to the
advantages it provides over those of conventional dosage routes,
such as oral and intravenous administration. For example,
biologically active substances or drugs delivered transdermally
avoids deactivation caused by pH and digestive enzymes upon passage
of the active substance through the gastrointestinal (GI) tract. In
addition, other advantages of transdermal delivery include, but are
not limited to, single application regimens or decreased dosages,
increased patient compliance, high percentage of drug reaching the
systemic circulation, sustained activity for drugs having short
half-lives, controlled release of drugs (no "burst effect"),
ability to quickly terminate drug dosing causing adverse effects
and administration of drugs without hypodermic injection.
[0004] The success of transdermal delivery in a mammal relies on
the ability of biologically active substances to penetrate the
outer layer of the epidermis known as the stratum corneum. The
stratum corneum is comprised mainly of about 10 to about 20 layers
of flattened dead cells (corneocytes) filled with keratin. Lipids,
such as free fatty acids, cholesterol, and ceramides, connect the
regions between the keratinized cells, forming a brick and
mortar-like structure. In mammals, this structure primarily serves
as a barrier to chemicals and biological agents, including
bacteria, fungus, and viruses.
[0005] The penetration of biologically active substances through
the stratum corneum occurs by either passive or active transport
mechanisms. Passive delivery or diffusion relies on a concentration
gradient between the drug at the outer surface and the inner
surface of the skin. The diffusion rate is proportional to the
gradient and is modulated by a molecule's size, hydrophobicity,
hydrophilicity and other physiochemical properties as well as the
area of the absorptive surface. Examples of passive delivery
systems include transdermal patches for controlled delivery of, for
example, nitroglycerine (angina), scopolamine (motion sickness),
fentanyl (pain control), nicotine (smoking cessation), estrogen
(hormone replacement therapy), testosterone (male hypogonadism),
clonidine (hypertension), and lidocaine (topical anesthesia). The
controlled delivery of these drugs can include the use of polymer
matrices, reservoirs containing drugs with rate-controlling
membranes and drug-in-adhesive systems.
[0006] In contrast, active delivery relies on ionization of the
drug or other pharmacologically active substances and on means for
propelling the charged ions through the skin. The rate of active
transport varies with the method used to increase movement and
propulsion of molecules, but typically this transport provides a
faster delivery of biologically active substances than that of
passive diffusion. Active transport delivery systems include
methods such as iontophoresis, sonophoresis, thermal microporation,
and microporation using mechanical means, such as microinjection
using microneedles or needleless injection.
[0007] Iontophoresis is a technique used to guide one or more
therapeutic ions in solution into the tissues and blood vessels of
the body by means of a galvanic or direct electrical current
supplied to wires that are connected to skin-interfacing
electrodes. Although ionotophoresis provides a method for
controlled drug delivery, irreversible skin damage can occur from
galvanic and pH burns resulting from electrochemical reactions that
occur at the electrode and skin interface. This reaction precludes
the use of this method when extended application times are needed
to achieve prolonged systemic effects.
[0008] Sonophoresis is another active transport method that uses
ultrasound varying in frequency from 20 kHz to 16 MHz to transport
substances across the stratum corneum. Sonophoresis affects
biological tissues by three main routes--thermal, cavitational and
acoustic streaming. For example, ultrasound will increase the
temperature of a given medium, and the absorption coefficient of
that medium increases proportionally with ultrasound frequency.
Cavitation can occur when ultrasound-induced pressure variation
causes rapid growth and collapse of gas bubbles, causing structural
alteration of the skin. Acoustic streaming, a phenomenon that
affects surrounding tissue structure, can occur when shear stresses
result from ultrasound reflections, distortions, and oscillations
of cavitation bubbles. It has also been postulated that ultrasound
interacts with the ordered lipids comprising the stratum corneum,
forming an opening for drug passage. The interruption of the
connective layer by any of the above-identified routes can lead to
an area of skin that is predisposed to sloughing as well as
bacterial and viral infiltration.
[0009] Microporation is an active transport method used to produce
micropores in the stratum corneum. Microporation is accomplished by
various means, including ablating the stratum corneum by local
rapid heating of water, puncturing the stratum corneum with a
micro-lancet calibrated to form a specific pore diameter, ablating
the stratum corneum by focusing a tightly focused beam of sonic
energy, hydraulically puncturing the stratum corneum with a high
pressure fluid jet, and puncturing the stratum corneum with short
pulses of electricity. Laser energy can also be used to cause
microporation. Although the diameter of the hole can be controlled,
microporation can cause irritation, damage and/or removal of
stratum corneum cells.
[0010] Because of the inherent problems of the above-identified
methods, a need exists for a safe and efficient transdermal drug
delivery that eliminates side-effects and damage to the barrier
function or appearance of the skin caused by drug administration.
It would therefore be desirable to provide compositions, methods,
and apparatuses to address these problems.
SUMMARY OF THE INVENTION
[0011] The problems associated with active intradermal or
transdermal delivery of biologically active substances can be
overcome by this invention, which relates to novel compositions,
methods, and devices for photokinetic intradermal, transdermal, or
transmembrane delivery of biologically active substances into or
through the stratum corneum or a biological membrane without
causing damage to this layer or underlying tissues, and without
denaturation and/or degradation of the biologically active
substance being administered.
[0012] The compositions, methods, and devices described herein
preferably use pulsed incoherent light to focus and deliver
biologically active substances through the outer most surface of
the skin, and either locally into the epidermis layer of the skin
(e.g., in cosmetic uses), or to an underlying tissue or blood
vessel (e.g., in pharmaceutical uses), or from an extracellular
environment to an intracellular environment. The depth of the
delivery can be fine tuned or controlled to achieve the intended
purpose. The intradermal drug deposition amount and transdermal
flux rate can be predicted and controlled by modulation of the
light energy. In some embodiments, compositions containing only
biologically active substances are used as delivery media, whereas
in other embodiments, biologically active substances used in
combination with other components are used as delivery media.
[0013] Methods and devices employing pulsed incoherent light are
used to actively transport a biologically active medium through the
outer surface of the skin or cell membrane. This provides many
advantages, including the ability to create a pathway for drug
delivery without causing damage to the skin or membrane while being
able to excite biologically active molecules without degrading or
denaturing them. In addition, the rate of delivery of the
biologically active component can be controlled, sustained, or
substantially stopped, by modulating the wavelength, pulse rate,
duty cycle and intensity of the light being used to
photokinetically propagate the component into or through the skin
or membrane. Finally, the use of a light pad containing more than
one light source permits light to expose a biologically active
medium over a well-defined surface area. The skin permeability can
be enhanced through the use of compositions, methods and devices
described herein.
[0014] Described herein is a novel platform technology pertaining
to enhanced permeation kinetics of a compound into and through a
tissue by the application of selected pulsed incoherent light
(photokinetic method). The invention described herein has shifted
the established tissue permeation paradigms away from the known
limitations of established fundamental axioms, such as molecular
weight limited permeation. This new photokinetic facilitated
permeation technology allows for administration of a wide range of
compounds having a wide range of molecular weights into and through
intact tissues, without damage to the tissues or chemical changes
to the molecule being delivered.
[0015] Therefore, one salient feature of the methods and systems of
the invention is its superior safety feature, which can be
particularly valuable for cosmetic applications. For example, since
the penetration distance is a function of the time exposed to the
light source (e.g., LED), the ability to regulate the flux rate of
the biologically active substance, including the ability to target
the delivery of the substance at a specific depth within the skin
or epidermis, enables targeted delivery of certain cosmetically
active substances to skin (without causing damages to the skin or
leading to unpredictable effects resulting from systemic delivery
of the substance).
[0016] Another safety feature of the subject delivery method
resides in the fact that the delivery method does not damage human
skin, and the active ingredients being delivered are not
transformed to potentially harmful substances by the light
energy.
[0017] Furthermore, using the subject dermal patch confers an
additional layer of safety, since the subject dermal patch is
produced from safe ingredients, many of which are natural, and
contains no preservatives or other potentially harmful substances
to the skin. The combination of the subject intradermal delivery
technology with the subject dermal patch not only unexpectedly
enhances delivery of the active ingredient, but also provides the
above safety features valued in cosmetic industry.
[0018] Human skin possesses remarkably different drug permeation
characteristics compared to animal skin. Animal skin permeation
testing has little relevance in predicting human skin drug
permeation. Partly because of the superior safety features of the
invention and the irrelevance of animal skin permeation testing, no
animal testing is needed for cosmetic products to be delivered to
human through the subject methods and patches.
[0019] Thus, in one respect, the invention provides a method for
photokinetic intradermal and/or transdermal delivery of a
biologically active substance having a molecular weight of no less
than 5800 Da to a subject, the method comprising: (1) preparing a
formulation comprising the biologically active substance; (2)
applying said formulation to a cellular surface on the skin of the
subject; (3) illuminating said formulation on said cellular surface
with a pulsed incoherent light having a selected wavelength, pulse
rate and duty cycle; and, (4) allowing said biologically active
substance in said formulation to permeate said cellular surface,
thereby effecting photokinetic intradermal and/or transdermal
delivery of the biologically active substance.
[0020] In certain embodiments, the biologically active substance
has a molecular weight of no less than 10 kDa, 25 kDa, 50 kDa, 100
kDa, 150 kDa, 200 kDa, 250 kDa, 300 kDa, 350 kDa, 400 kDa, 500 kDa,
600 kDa, 700 kDa, 800 kDa, 900 kDa, 1,000 kDa, 1,500 kDa, 1,600
kDa, 2,000 kDa, 2,200 kDa, 2,500 kDa, or 3,000 kDa.
[0021] In certain embodiments, the subject is a human.
[0022] In certain embodiments, such as in pharmaceutical
application, the subject is not a rodent (e.g., one or more of
mouse, rat, squirrel, porcupine, beaver, chipmunk, guinea pig,
vole, etc.).
[0023] In certain embodiments, the skin is non-porated intact skin,
or the skin is porated by chemical, electrical, and/or physical
means.
[0024] In certain embodiments, the formulation comprises a solution
and a solvent.
[0025] In certain embodiments, the solvent is an aqueous or an
organic solvent.
[0026] In certain embodiments, the aqueous solvent is an aqueous
solution of ethyl lactate and proplyene glycol, water, or an
aqueous solution of ethyl lactate or propylene glycol.
[0027] In certain embodiments, the formulation comprises a gelling
agent.
[0028] In certain embodiments, the gelling agent comprises a
hydroxyethyl cellulose, a cellulose derivative, a pectine, an agar,
an alginic acid or a salt thereof, a guar gum, a polyvinyl alcohol,
a polyethylene oxide, a propylene carbonate, a polyethylene glycol,
a hexylene glycol sodium carboxymethylcellulose, a polyacrylate, a
polyoxyethylene-polyoxypropylene, a block copolymer, a pluronics, a
wood wax alcohol, a tyloxapol, and/or a hyaluronic acid (e.g., one
with an average molecular weight of about 500 kDa, 600 kDa, 700
kDa, 800 kDa, 900 kDa, 1000 kDa, 1200 kDa, 1400 kDa, or 1,600-2,200
kDa).
[0029] In certain embodiments, the formulation further comprises a
photocatalytic agent.
[0030] In certain embodiments, the photocatalytic agent has a band
gap energy of between about 2.9 eV and about 3.2 eV, is a rutile
form of titanium dioxide, or is an anatase form of titanium
dioxide. For cosmetic applications, UV filters do not penetrate the
skin.
[0031] In certain embodiments, the biologically active substance
comprises a chemical, an antibiotic, a hormone, a peptide, an
antibody, a protein, a plant extract (such as those used in the
cosmetic industry), or a mixture thereof.
[0032] In certain embodiments, the biologically active substance
comprises a drug.
[0033] In certain embodiments, the drug is a cytotoxic drug.
[0034] In certain embodiments, the drug comprises or further
comprises an analgesic, an anaesthetic, an antacid, an antianxiety
drug, an antiarrhythmics, an antibacterial, an antibiotic, an
anticoagulant and thrombolytic, an anticonvulsants, an
antidiarrheals, an antiviral, a barbiturate, and/or a vitamin.
[0035] In certain embodiments, the biologically active substance
comprises or further comprises chemicals and said chemicals
comprise a polar or a non-polar compound.
[0036] In certain embodiments, the polar compound is selected from
the group consisting of theophylline-7 acetic acid, sodium ascorbyl
phosphate, ascorbic acid, ascorbyl palmitate, pyridoxine, nicotinic
acid, and lidocaine.
[0037] In certain embodiments, the non-polar compound is selected
from the group consisting of theobromine, theophylline, caffeine,
and nicotinamide.
[0038] In certain embodiments, the biologically active substance
comprises sodium hyaluronate (such as one with a M.W. of no less
than 20 kDa, 50 kDa, 100 kDa, 200 kDa, 400 kDa, 500 kDa, 1000 kDa,
1250 kDa, 1500 kDa, 1600 kDa, 2000 kDa, 2100 kDa, 2200 kDa, 2500
kDa, 3000 kDa, 3500 kDa, 4000 kDa, 4500 kDa, 5000 kDa or more).
[0039] In certain embodiments, the biologically active substance is
a cosmetic agent, or a therapeutic agent.
[0040] In certain embodiments, the biologically active substance
comprises or further comprises: (1) a peptide selected from the
group consisting of Gly-Tyr, Val-Tyr-Val, Tyr-Gly-Gly-Phe-Met (SEQ
ID NO: 1), Tyr-Gly-Gly-Phe-Leu (SEQ ID NO: 2), and
Asp-Arg-Val-Tyr-Ile-His-Pro-Phe (SEQ ID NO: 3); (2) a hormone
selected from the group consisting of methionine enkephalin
acetate, leucine enkephalin, angiotensin II acetate,
.beta.-estradiol, methyl testosterone, and progesterone; or, (3) a
protein selected from the group consisting of enzymes, non-enzymes,
antibodies, and glycoproteins.
[0041] In certain embodiments, the cellular surface is a cell
membrane.
[0042] In certain embodiments, the pulsed incoherent light is
selected from the group consisting of fluorescent, ultraviolet,
visible, near infrared, LED (light emitting diode), and halogen
light.
[0043] In certain embodiments, the fluorescent light has a
wavelength range from about 260 nm to about 760 nm.
[0044] In certain embodiments, the ultraviolet light has a
wavelength range from about 340 nm to about 900 mm.
[0045] In certain embodiments, the visible light has a wavelength
range from about 340 nm to about 900 nm.
[0046] In certain embodiments, the near infrared light has a
wavelength range from about 340 nm to about 900 nm.
[0047] In certain embodiments, the halogen light has a wavelength
range from about 340 nm to about 900 nm.
[0048] In certain embodiments, the wavelength is selected from the
group consisting of 350 nm, 390 nm, 405 nm, and 450 nm.
[0049] In certain embodiments, the pulse rate is between about 1.7
cycles per second (cps) and about 120 cps, or about 1.7 cps and
about 80 cps.
[0050] In certain embodiments, the pulse rate is between about
24-100 cps.
[0051] In certain embodiments, the duty cycle is between about 50%
and about 75%.
[0052] In other embodiments, a discrete On time and a discreet OFF
time, in the range of, for example, 1% ON/99% OFF to 99% ON/1% OFF
(including any integer values in between, such as 2% ON98% OFF, 5%
ON95% OFF, 10% ON90% OFF, 15% ON85% OFF, etc.) are all contemplated
embodiments of the invention.
[0053] In certain embodiments, the method further comprises
adjusting the transdermal flux rate of the biologically active
substance by modulating the light energy.
[0054] In certain embodiments, the method further comprises
adjusting the flux rate of the biologically active substance by
modulating the light energy, in order to deliver all or
substantially all biologically active substance intradermally
(within the skin).
[0055] In certain embodiments, the formulation comprises a dermal
patch, said dermal patch containing a red algae
polysaccharide-based matrix, wherein said red algae polysaccharide
is an extract of Chondrus crispus at 2% by weight of the dermal
patch.
[0056] In certain embodiments, the concentration (by weight) of
Chondrus crispus is about 0.5%, 1.0%, 1.5%, 2.0%, 2.5%, 3.0%, 3.5%,
or about 4%. Any two values may also serve at the lower and higher
ends of a suitable range. In certain embodiments, the concentration
(by weight) of Chondrus crispus is about 1.5%-2.5%, preferably
2.0%.
[0057] In certain embodiments, such as in cosmetic application, the
dermal patch is disposable (e.g., after single use or limited time
of uses), biodegradable, environmentally safe, and/or produced
using essentially natural ingredients. Preferably, the biologically
active substance (e.g., the cosmetic active ingredient) is present
in the dermal patch at a higher amount (e.g., 20-fold excess,
10-fold excess, 5-fold excess, etc.) compared to the amount to be
delivered, so as to maintain relatively constant rate of release of
the active ingredient.
[0058] In certain embodiments, the dermal patch further comprises
methylpropanediol and glycerine as moisturising and anti-microbial
ingredients.
[0059] In certain embodiments, the dermal patch further comprises
butylene glycol and sorbitol as moisturising ingredients.
[0060] In certain embodiments, the dermal patch comprises water,
methylpropanediol, glycerin, Chondrus crispus, and optionally a
preservative.
[0061] In certain embodiments, the dermal patch comprises (as
weight %) 67% water, 10% methylpropanediol, 20% glycerin, 2%
Chondrus crispus and optionally a preservative.
[0062] In certain embodiments, the dermal patch comprises water,
butylene glycol, sorbitol, Chondrus crispus and optionally a
preservative.
[0063] In certain embodiments, the dermal patch comprises (as
weight %) 67% water, 10% butylene glycol, 20% sorbitol, 2% Chondrus
crispus and optionally a preservative.
[0064] In certain embodiments, the dermal patch comprises no
preservative.
[0065] In certain embodiments, the dermal patch is prepared
according to a processing comprising the following steps: (1)
providing a water mixture of the vegetable matrix, excipients and
active ingredients; (2) heating the mixture to a temperature
between 30-90.degree. C.; (3) casting the mixture in a mould while
keeping the temperature between 30 and 90.degree. C.; and, (4)
cooling the mixture to about room temperature.
[0066] In certain embodiments, the mixture is heated at 90.degree.
C. under mixing.
[0067] In certain embodiments, in step (3), the temperature is
maintained at 50-60.degree. C.
[0068] In certain embodiments, the mixing step (1) is performed in
a planetary mixer equipped with a jacketing for heating under
controlled temperature.
[0069] In certain embodiments, in step (3), the mixture is casted
in a plastic mould for blisters.
[0070] In certain embodiments, the method further comprises the
application of a sheet of aluminium/polythene tie layer to seal the
blister and the die-cut of the blister to obtain the desired final
shape of the finished product.
[0071] In a related aspect, the invention provides a method for
photokinetic intradermal and/or transdermal delivery of a
biologically active substance to a subject, the method comprising:
(1) preparing a dermal patch comprising the biologically active
substance, wherein the dermal patch comprises a red algae
polysaccharide-based matrix, and wherein said red algae
polysaccharide is an extract of Chondrus crispus at 2% by weight of
the dermal patch; (2) applying said dermal patch to a cellular
surface on the skin of the subject; (3) illuminating said dermal
patch on said cellular surface with a pulsed incoherent light
having a selected wavelength, pulse rate and duty cycle; and, (4)
allowing said biologically active substance in said dermal patch to
permeate said cellular surface, thereby effecting photokinetic
intradermal and/or transdermal delivery of the biologically active
substance.
[0072] Specific embodiments of this aspect of the invention are
described above and not repeated verbatim here.
[0073] Another aspect of the invention provides a device for
photokinetic intradermal and/or transdermal delivery of a
biologically active substance having a molecular weight of no less
than 5800 Da to a subject, said device comprising: (1) a generator
that provides an oscillating electrical pulse; (2) at least one
light emitting diode that receives the oscillating electrical pulse
and responds by providing an incoherent light; and, (3) a donor
cell that holds a formulation comprising the biologically active
substance, wherein the donor cell is positioned to receive the
incoherent light.
[0074] Specific embodiments of this aspect of the invention are
described above and not repeated verbatim here.
[0075] Another aspect of the invention provides a device for
photokinetic intradermal and/or transdermal delivery of a
biologically active substance to a subject, said device comprising:
(1) a generator that provides an oscillating electrical pulse; (2)
at least one light emitting diode that receives the oscillating
electrical pulse and responds by providing an incoherent light;
and, (3) a donor cell that holds a dermal patch comprising the
biologically active substance, wherein the dermal patch comprises a
red algae polysaccharide-based matrix, wherein said red algae
polysaccharide is an extract of Chondrus crispus at 2% by weight of
the dermal patch, and wherein the donor cell is positioned to
receive the incoherent light.
[0076] In certain embodiments, the generator is a repeat cycle
square wave pulse generator.
[0077] In certain embodiments, the device further comprises a light
pad, wherein at least one light emitting diode is embedded in said
light pad.
[0078] In certain embodiments, the light pad is comprised of an
optically clear material.
[0079] In certain embodiments, the optically clear material is
poly(methylmethacrylate) or silicone rubber.
[0080] Other specific embodiments of this aspect of the invention
are described above and not repeated verbatim here.
[0081] It should be understood that any embodiments of the
invention described herein, including those described under
different aspects of the invention, can be combined with one or
more other embodiments of the invention.
BRIEF DESCRIPTION OF THE DRAWINGS
[0082] FIG. 1 shows a Franz skin diffusion device equipped with a
light source to generate pulses of defined wavelengths for testing
of biologically active substances.
[0083] FIG. 2 shows a Franz skin diffusion device similar to that
in FIG. 1 except the light source is embedded in an optically clear
medium or light pad that does not absorb the wavelength emitted
from the light source.
[0084] FIG. 3A shows an array of light sources embedded in an
optically clear medium or light pad and electrically coupled to
each other and to a control device or power supply.
[0085] FIG. 3B illustrates multiple light sources electrically
connected in series and embedded in an optically clear medium or
light pad wherein the upper surface of the light pad is coated with
a reflective layer and the lower surface of the light pad is
optically clear.
[0086] FIG. 4 illustrates exemplary Franz diffusion cell apparatus
for in vitro determination of photokinetic conditions of light
wavelength and pulse rate.
[0087] FIG. 5 is a schematic drawing showing determination of
intradermal drug concentration.
[0088] FIG. 6 shows molecular weight range of compounds delivered
into and through human skin using the photokinetic system.
Significant quantities were delivered when compared to passive
permeation control conditions. The photokinetic intradermal and
transdermal delivery system provided surprising and unexpected
results as the molecules tested herein are well beyond (>4000
times) the 500 Dalton upper molecular weight useful limit normally
associated with permeation into and through intact (non-porated)
skin.
[0089] FIG. 7 shows that IGF-1 was delivered into intact human skin
at significantly higher concentrations compared to passive
permeation, as measured at 15, 30 and 60 minutes.
[0090] FIG. 8 shows human growth hormone (HGH) transdermal
permeation quantities as measured at 24 hours exposed to 4
wavelengths of light pulsed at 24 cycles per second (24 cps).
[0091] FIG. 9A shows transdermal permeation of 69 kDa hyaluronic
acid, determined under photokinetic conditions of 390 nm, 405 nm,
436 nm, and 450 nm light pulsed at 24 cycles per second (cps) and
100 cps compared to passive permeation at 24 hours.
[0092] FIG. 9B shows intradermal deposition of 69 kDa hyaluronic
acid, determined under photokinetic conditions of 390 nm, 405 nm,
436 nm, and 450 nm light pulsed at 24 cycles per second (cps) and
100 cps compared to passive deposition at 24 hours.
[0093] FIG. 10A shows transdermal permeation of botulinum toxin
type A at 24 hours of photokinetic conditions of light wavelengths
405 nm and 450 nm pulsed at 24 cps and 100 cps compared to passive
control. The four photokinetic conditions all produced significant
increases in transdermal flux compared to passive permeation
(p<0.001).
[0094] FIG. 10B shows intradermal deposition of botulinum toxin
type A at 24 hours of photokinetic conditions of light wavelengths
405 nm and 450 nm pulsed at 24 cps and 100 cps compared to passive
control. The photokinetic conditions of 450 nm light pulsed at 24
and 100 cps both produced significant increases in achievable
tissue deposition of botulinum toxin type A compared to passive
tissue deposition (p<0.05).
[0095] FIG. 11A shows transdermal permeation of 2,100 kDa
hyaluronic acid determined under photokinetic conditions of 390 nm,
405 nm, 436 nm, and 450 nm light pulsed at 24 cycles per second
(cps) and 100 cps compared to passive permeation at 24 hours.
[0096] FIG. 11B shows intradermal deposition of 2,100 kDa
hyaluronic acid determined under photokinetic conditions of 390 nm,
405 nm, 436 nm, and 450 nm light pulsed at 24 cycles per second
(cps) and 100 cps compared to passive deposition at 24 hours.
[0097] FIG. 12 shows intradermal deposition of hyaluronic acid
formulated in a gel (1% HA), a cream (1% HA) and patch (0.5% HA) at
15, 30 and 60 minutes.
DETAILED DESCRIPTION OF THE INVENTION
1. Overview
[0098] Optimum therapeutic outcomes require not only proper drug
selection but also effective drug delivery.
[0099] Oral delivery (e.g. pills) has been considered as the most
appropriate method of drug administration for decades. Most of the
drugs that cannot be taken by oral delivery have traditionally been
administered by injections with hypodermic needles. However,
hypodermic injections have many disadvantages, such as pain,
potential infections and the requirement for medical expertise to
complete the injection process (Park et al. 2005).
[0100] The human skin is also a readily accessible surface for drug
delivery. Over the past three decades, developing controlled drug
delivery has become increasingly important in the pharmaceutical
industry. The pharmacological response, both the desired
therapeutic effect and the undesired adverse effect, of a drug is
dependent on the concentration of the drug at the site of action,
which in turn depends upon the dosage form and the extent of
absorption of the drug at the site of action.
[0101] Skin of an average adult body covers a surface of
approximately 2 m.sup.2, and receives about one-third of the blood
circulating through the body. Skin contains an uppermost layer,
epidermis which has morphologically distinct regions; basal layer,
spiny layer, stratum granulosum and upper most stratum corneum, it
consists of highly cornified (dead) cells embedded in a continuous
matrix of lipid membranous sheets. These extracellular membranes
are unique in their compositions and are composed of ceramides,
cholesterol and free fatty acids. The human skin surface is known
to contain, on an average, 10-70 hair follicles and 200-250 sweat
ducts on every square centimeter of the skin area. It is one of the
most readily accessible organs of the human body. The potential of
using the intact skin as the port of drug administration to the
human body has been recognized for several decades, but skin is a
very difficult barrier to the ingress of materials allowing only
small quantities of a drug to penetrate over a period of time.
[0102] Facilitated or "active" transdermal methods wherein
electrical (iontophoresis) or sound energy (phonophoresis or
sonophoresis) is added into the system have been employed as
methods to increase the drug flux across the skin. In the absence
of skin poration side effects with these methods (electroporation
caused by excessive electrical energy or destructive micro-bubble
formation within the epidermis from cavitation induced by sound
energy) the drug flux across intact skin remains limited by the
drug molecular weight. It is widely accepted and understood by
those familiar with the art that there is a practical upper
molecular weight limit for drugs permeating though intact human
skin. This upper molecular weight limit is considered to be about
500 Daltons (Da) with considerable diminished permeation in drugs
with larger molecular weights. This molecular weight limiting
property is commonly referred to in the art as the "500 Dalton
Rule" wherein as the molecular weight of the drug increases the
expected permeation through human skin decreases and generally
falls to insignificant amounts as the molecular weight approaches
500 Da.
[0103] Transdermal drug delivery (TDD)--the delivery of drugs
across the skin and into systemic circulation--is distinct from
topical drug penetration or intradermal delivery, which targets
local areas, such as delivery or application of cosmetic
ingredients to the skin. Transdermal drug delivery takes advantage
of the relative accessibility of the skin, and is an alternative
route of drug administration to pills and injections. This method
operates by delivering drugs into the human body across the skin
using devices such as a patch (e.g., a transdermal patch). The
transdermal patches have the ability to eliminate at least some of
the problems mentioned above. They usually contain a drug reservoir
that can maintain a steady drug flow of up to about one week
(Prausnitz et al., 2004). Although these patches have proven to be
very successful, they depend on the characteristics of the drug,
e.g., the size, charge, and even some physiochemical properties
(Naik et al., 2000) to be successful. This is largely due to the
barrier function of the skin represented by the outer layer of the
skin, the stratum corneum, which generally allows diffusion of only
small molecular weight solutes (less than 500 Da), with the ability
to allow penetration of certain oil-soluble solutes (Shah 2003). To
circumvent this diffusion limitation, methods have been developed
to more effectively deliver drugs across the stratum corneum,
including chemical enhancers (Williams & Barry 2004) or
physical enhancer techniques, e.g., iontophoresis (Kalia et al.
2004) and ultrasound (Prausnitz et al. 2004). However, the high
cost, complexity, and the difficulty in dealing with these methods
at home pose problems for potential users.
[0104] Transdermal drug delivery offers several important
advantages over more traditional dosage forms. The steady
permeation of drug across the skin allows for more consistent serum
drug levels, often a goal of therapy. Intravenous infusion also
achieves consistent plasma levels, but it is more invasive than
transdermal drug delivery.
[0105] The lack of peaks in plasma concentration can reduce the
risk of side effects. Thus, drugs that require relatively
consistent plasma levels are very good candidates for transdermal
drug delivery. In addition, if toxicity were to develop from a drug
administered transdermally, the effects could be limited by
removing the patch.
[0106] Another advantage is convenience, especially notable in
patches that require only once weekly application. Such a simple
dosing regimen can aid in patient adherence to drug therapy.
Transdermal drug delivery can be used as an alternative route of
administration to accommodate patients who cannot tolerate oral
dosage forms. It is of great advantage in patients who are
nauseated or unconscious. Drugs that cause gastrointestinal upset
can be good candidates for transdermal delivery because this method
avoids direct effects on the stomach and intestine. Drugs that are
degraded by the enzymes and acids in the gastrointestinal system
may also be good targets. First pass metabolism, an additional
limitation to oral drug delivery, can be avoided with transdermal
administration. Thus, in certain embodiments, the drugs/molecules
to be delivered using the instant methods and systems are such
drugs, or are intended to be delivered to such patient
populations.
[0107] Other advantage includes suitability for using with drug
candidates with short half-life and low therapeutic index.
[0108] The first TDD System (TDDS) was developed for scopolamine
for motion sickness in 1981. Since then, many TDDS have appeared in
market with great success. In spite of the therapeutic success
achieved in last 28 years by using TDDS, the number of TDDS
available in the market place is comparatively very few. This is
mainly due to inherent limitations of the TDDS listed below.
[0109] One of the greatest disadvantages to transdermal drug
delivery is the possibility that a local irritation may develop at
the site of application. Erythema, itching, and local edema can be
caused by the drug, the adhesive, or other excipients in the patch
formulation. For most patients, site rotation can minimize
irritation. However, some patients develop severe allergic
reactions to transdermal patches, and, in these cases, therapy must
be discontinued.
[0110] Another significant disadvantage of transdermal drug
delivery is that the skin's low permeability limits the number of
drugs that can be delivered in this manner. Because the skin serves
protective functions, it inhibits compounds from crossing it. Many
drugs with a hydrophilic structure permeate the skin too slowly to
be of therapeutic benefit. Drugs with a lipophillic character,
however, are better suited for transdermal delivery.
[0111] In order to maintain consistent release rates, transdermal
patches may contain a surplus of active molecule. A stable
concentration gradient is the mechanism used to maintain consistent
release rates and constant serum drug levels. Most transdermal
patches contain 20 times the amount of drug that will be absorbed
during the time of application. Thus, after removal, most patches
contain at least 95% of the total amount of drug initially in the
patch. Therefore; patients must exercise care when disposing of
patches. For example, each patch should be folded in half and the
adhesive sides should be stuck together. As an additional
precaution, patches may be flushed down the toilet rather than
discarded in household trash, where children and pets may find them
and ingest the remaining drug.
[0112] Damage to a transdermal patch, particularly a membrane or
reservoir patch, can result in poor control over the release rate.
The release rate from a damaged patch would more likely be
controlled by the skin than the patch, resulting in a higher,
perhaps toxic, rate of drug delivery. Patients should be advised to
discard a patch if the outer packaging or the patch itself appears
damaged or altered in any way.
[0113] Other limitations for TDDS include factors that limit a drug
candidate's ability to be incorporated into a transdermal delivery
system, such as: higher molecular weight (e.g., >500 Da) that
renders the drug molecule harder to penetrate the stratum corneum;
very low or high partition coefficient for certain drugs, which
prevents such drugs to reach systemic circulation; and high melting
drugs due to their low solubility both in water and fat. Such
candidate drugs with one or more of these properties may not be
efficiently delivered across the skin without effectively making
suitable modifications in the conventional transdermal delivery
systems.
[0114] There are two concepts in the design of transdermal
delivery, namely, the reservoir type and the matrix type. Others
are actually extensions of these two concepts and both involve
diffusion of drug molecule from the topically applied donor
reservoir and into and through the skin barrier.
[0115] Modulation of formulation excipients and addition of
chemical enhancers, such as fatty acids, surfactants, esters and
alcohols that exert their action via a temporary alteration of
barrier properties of the stratum corneum by various mechanisms,
including enhancing solubility, partitioning the stratum corneum,
fluidizing the crystalline structure of the stratum corneum and
causing dissolution of stratum corneum lipids can enhance drug
flux. However, due to low permeability coefficients of
macromolecules, the enhancement effects required to ensure delivery
of pharmacologically effective concentrations are likely to be
beyond the capability of chemical enhancers tolerated by the skin.
Therefore, several new active transport technologies have been
developed for the transdermal delivery of "troublesome" drugs as
the development of modified novel physical techniques have overcome
the limitations of chemical enhancement techniques.
[0116] In transdermal technology, emphasis is placed on producing
therapeutic drug flux rates using a limited skin surface area with
minimal skin irritating or damaging side effects. Applicants have
previously described a method of enhanced or facilitated
transdermal drug delivery called "photokinetic" drug delivery (see
US2004-0131687A1, U.S. Pat. Nos. 7,458,982 and 7,854,753, and EP
1556061 B1, including all examples and drawings thereof, are all
incorporated herein by reference) for delivery of biologically
active substances using pulsed incoherent light, wherein the
application of pulsed incoherent light onto a drug on the surface
of intact skin allows or facilitates the permeation of that drug
into and through the skin.
[0117] More than 80% of all traded drugs have molecular weights
below 450 Da. There are therapeutic agents with larger molecular
weights, but these agents are generally considered well beyond the
accepted known limits of molecular-weight-limited skin permeation.
Against this background, and to the complete surprise and utter
amazement of the Applicants, it was found that the photokinetic
system can be utilized for skin permeation (e.g., intradermal and
transdermal delivery) of large to very large molecular weight
drugs, such as drugs in the molecular weight range of 7,676 Da for
insulin like growth factor-1 (IGF-1) up to and including 2,180,000
Da (2180 kDa) hyaluronic acid (HA), into and through intact skin.
This result is surprising and completely unexpected, partly because
the increase in molecular weight limits for intra- and transdermal
drug delivery are several thousand times more than the widely
accepted 500 Da upper molecular weight limit.
[0118] Thus one aspect of the invention provides method for
photokinetic intradermal and/or transdermal delivery of a
biologically active substance having a molecular weight of no less
than 5800 Da to a subject, the method comprising: (1) preparing a
formulation comprising the biologically active substance; (2)
applying said formulation to a cellular surface on the skin of the
subject; (3) illuminating said formulation on said cellular surface
with a pulsed incoherent light having a selected wavelength, pulse
rate and duty cycle; and, (4) allowing said biologically active
substance in said formulation to permeate said cellular surface,
thereby effecting photokinetic intradermal and/or transdermal
delivery of the biologically active substance.
[0119] In certain embodiments, the biologically active substance
has a molecular weight of no less than 10 kDa, 25 kDa, 50 kDa, 100
kDa, 150 kDa, 200 kDa, 250 kDa, 300 kDa, 350 kDa, 400 kDa, 500 kDa,
600 kDa, 700 kDa, 800 kDa, 900 kDa, 1,000 kDa, 1,500 kDa, 1,600
kDa, 2,000 kDa, 2,200 kDa, 2,500 kDa, or 3,000 kDa.
[0120] Another aspect of the invention is partly based on the
unexpected results arising from combining the photokinetic drug
delivery method/system with a dermal patch described in WO
09/124763 A2.
[0121] Typically, when drugs are applied to the skin, they may be
formulated in a liquid or gel form or in some physical containment
method such as the common transdermal patch. Also, typically the
topical application method may have an effect on the permeation of
the drug from the topical application and into the skin.
Micro-emulsion drug compositions generally provide better
transdermal flux than water based gels or drug/patch
combinations.
[0122] Applicants have examined the combination effects of a
particular drug patch (described in WO 09/124763 A2) containing
2,180 kDa hyaluronic acid with photokinetic technology, and have
found that this combination has surprisingly better permeation
results (e.g., up to an increase of about 200-300%) when compared
with the same drug in a gel or emulsion formulation.
[0123] Thus another aspect of the invention provides method for
photokinetic intradermal and/or transdermal delivery of a
biologically active substance to a subject, the method comprising:
(1) preparing a dermal patch comprising the biologically active
substance, wherein the dermal patch comprises a red algae
polysaccharide-based matrix, and wherein said red algae
polysaccharide is an extract of Chondrus crispus at 2% by weight of
the dermal patch; (2) applying said dermal patch to a cellular
surface on the skin of the subject; (3) illuminating said dermal
patch on said cellular surface with a pulsed incoherent light
having a selected wavelength, pulse rate and duty cycle; and, (4)
allowing said biologically active substance in said dermal patch to
permeate said cellular surface, thereby effecting photokinetic
intradermal and/or transdermal delivery of the biologically active
substance.
[0124] In a related aspect, the invention also provides a device
for photokinetic intradermal and/or transdermal delivery of a
biologically active substance having a molecular weight of no less
than 5800 Da to a subject, said device comprising: (1) a generator
that provides an oscillating electrical pulse; (2) at least one
light emitting diode that receives the oscillating electrical pulse
and responds by providing an incoherent light; and, (3) a donor
cell that holds a formulation comprising the biologically active
substance, wherein the donor cell is positioned to receive the
incoherent light.
[0125] In yet another related aspect, the invention provides a
device for photokinetic intradermal and/or transdermal delivery of
a biologically active substance to a subject, said device
comprising: (1) a generator that provides an oscillating electrical
pulse; (2) at least one light emitting diode that receives the
oscillating electrical pulse and responds by providing an
incoherent light; and, (3) a donor cell that holds a dermal patch
comprising the biologically active substance, wherein the dermal
patch comprises a red algae polysaccharide-based matrix, wherein
said red algae polysaccharide is an extract of Chondrus crispus at
2% by weight of the dermal patch, and wherein the donor cell is
positioned to receive the incoherent light.
[0126] More detailed aspects and embodiments of the inventions are
set forth herein, including in the examples.
2. Definitions
[0127] Unless otherwise defined herein, scientific and technical
terms used in connection with the invention shall have the meanings
that are commonly understood by those of ordinary skill in the art.
Further, unless otherwise required by context, singular terms shall
include pluralities and plural terms shall include the singular.
Generally, nomenclatures used in connection with, and techniques
of, column chromatography, optics, chemistry, peptide and protein
chemistries, nucleic acid chemistry and molecular biology described
herein are those well known and commonly used in the art.
[0128] The following terms, unless otherwise indicated, shall be
understood to have the following meanings:
[0129] The term "biologically active substance" refers generally to
any chemical, drug, antibiotic, peptide, hormone, protein, DNA, RNA
and mixtures thereof that affects biological pathways or interacts
with cellular components. In certain embodiments, the biologically
active substance comprises one or more cosmetic active
ingredients.
[0130] The term "chemical" denotes any naturally found or
synthetically made or extracted small molecule or polymer
(including naturally extracted cosmetic ingredients or cosmetic
ingredients extracted from a natural source, such as polyphenols,
plant extracts, etc.). In certain embodiments, a chemical may also
include certain biofermentation compounds (such as sodium
hyaluronate, etc.).
[0131] A chemical can be a polar (hydrophilic), non-polar
(hydrophobic), oleophobic or oleophilic compound. Although not an
exhaustive list, examples of polar compounds include polyphenols,
phytosterols, theophylline-7-acetic acid, sodium ascorbyl
phosphate, ascorbic acid, ascorbyl palmitate, pyridoxine, nicotinic
acid and lidocaine. Examples of non-polar compounds include
theobromine, theophylline, caffeine and nicotinamide. Oleophobic
compounds are those compounds lacking affinity for oils and
oleophilic compounds are any compounds that have a stronger
affinity for oils over that of water. Accordingly, the invention
described herein is particularly useful for transport of compounds
with chromophores, which can be polar, non-polar, oleophobic,
including fluorochemicals, and oleophilic.
[0132] The term "drug" denotes any natural or synthetic compound
used for therapeutic treatment in mammals. Examples of drugs
include, but are not limited to, analgesics, antacids, antianxiety
drugs, antiarrhythmics, antibacterials, antibiotics, anticoagulants
and thrombolytics, anticonvulsants, antidepressants,
antidiarrheals, antiemetics, antifungals, antihistamines,
antihypertensives, anti-inflammatories, antieoplastics,
antipsychotics, antipyretics, antivirals, barbiturates,
beta-blockers, bronchodilators, cold cures, corticosteroids, cough
suppressants, cytotoxics, decongestants, diuretics, expectorant,
hormones, hypoglycemic s, immunosuppressives, laxatives, muscle
relaxants, sedatives, sex hormones, sleeping drugs, tranquilizer
and vitamins.
[0133] Vitamins are organic chemicals that are essential for
nutrition in mammals and are typically classified as fat-soluble or
water-soluble. Vitamins required to maintain health in humans
include, but are not limited to, vitamin A (retinol), precursor to
vitamin A (carotene), vitamin B.sub.1 (thiamin), vitamin B.sub.2
(riboflavin), vitamin B.sub.3 (nicotinic acid), vitamin B
(pantothenic acid), vitamin C (ascorbic acid), vitamin D
(calciferol), vitamin E (tocopherol), vitamin H (biotin) and
vitamin K (naphtoquinone derivatives).
[0134] The term "antibiotic" refers to any natural or synthetic
substance that inhibits the growth of or destroys microorganisms in
the treatment of infectious diseases. Although not an exhaustive
list, examples of antibiotics include amoxycillin, ampicillin,
penicillin, clavulanic acid, aztreonam, imipenem, streptomycin,
gentamicin, vancomycin, clindamycin, ephalothin, erythromycin,
polymyxin, bacitracin, amphotericin, nystatin, rifampicin,
teracycline, coxycycline, chloramphenicol and zithromycin.
[0135] The term "peptide" refers to a compound that contains 2 to
50 amino acids and/or imino acids connected to one another. The
amino acids can be selected from the 20 naturally occurring amino
acids. The twenty conventional amino acids and their abbreviations
follow conventional usage. See Immunology--A Synthesis (2.sup.nd
Edition, E. S. Golub and D. R. Gren, Eds., Sinauer Associates,
Sunderland, Mass. (1991)), which is incorporated herein by
reference. The amino acids can also be selected from non-natural
amino acids such as those found at the Sigma-Aldrich web site,
including without limitation: .beta.-amino acids (b.sup.3 and
b.sup.2), homo-amino acids, cyclic amino acids, aromatic amino
acids, Pro and Pyr derivatives, 3-substituted Alanine derivatives,
Glycine derivatives, ring-substituted Phe and Tyr Derivatives,
linear core amino acids, diamino acids (see Sigma-Aldrich ChemFiles
Vol. 1 No. 5 (Unnatural Amino Acids); Vol. 2 No. 4 (Unnatural Amino
Acids II); and Vol. 4 No. 5 (Unnatural Amino Acids: Tools for Drug
Discovery). Although not an exhaustive list, examples of peptides
include glycine-tyrosine, valine-tyrosine-valine,
tyrosine-glycine-glycine-phenylalanine-methionine,
tyrosine-glycine-glycine-phenylalanine-leucine and aspartic
acid-arginine-valine-tyrosine-isoleucine-histidine-proline-phenylalanine.
[0136] "Preservative(s)" as used herein include the ones listed in
the following web site
http://ec.europa.eu/consumers/cosmetics/cosing/index.cfm?fuseaction=searc-
h.results &annex_v1=VI&search. A PDF copy of which is
attached as appendix. Also see European Cosmetics Directive
76/768/EEC--Annex VI part 1 ("list of preservatives allowed" from
"List of preservatives which cosmetic products may contain").
[0137] Representative preservatives include: parabens (methyl-,
ethyl-, propyl-, butylparaben); urea-derivatives (imidazolidinyl
urea, diazolidinyl urea); isothiazolones (methylchloro-,
methyl-isothiazolinone); halogen-organic actives (iodopropynyl
butylcarbamate, methyldibromo glutaronitrile); organic acids &
others (sodium benzoate, chloracetamide, EDTA, phenoxyethanol,
triclosan, DMDM-hydantoin, quaternium-15).
[0138] In certain embodiments, preservatives may also comprise
certain natural components that reinforce the anti-microbial
efficacy, such as extracts (grapefruit seed, rosemary); essential
oils (tea tree, neem seed, thyme); vitamins (vitamin E, vitamin C),
alcohols, glycols, or glycerin, etc. In other embodiments,
preservatives do not include such natural components.
[0139] The term "hormone" refers to a substance that originates in
an organ, gland, or part, which is conveyed through the blood to
another part of the body, stimulating it by chemical action to
increase functional activity or to increase secretion of another
hormone. Although not an exhaustive list, examples of hormones
include methionine enkephalin acetate, leucine enkephalin,
angiotensin II acetate, .beta.-estradiol, methyl testosterone,
progesterone and insulin.
[0140] A polypeptide is defined as a chain of greater than 50 amino
acids and/or imino acids connected to one another.
[0141] A protein is a large macromolecule composed of one or more
polypeptide chains. The term "isolated protein" is a protein that
by virtue of its origin or source of derivation (1) is not
associated with naturally associated components that accompany it
in its native state, (2) is free of other proteins from the same
species, (3) is expressed by a cell from a different species, or
(4) does not occur in nature. Thus, a protein that is chemically
synthesized or synthesized in a cellular system different from the
cell from which it naturally originates will be "isolated" from its
naturally associated components. A protein may also be rendered
substantially free of naturally associated components by isolation,
using protein purification techniques well known in the art.
[0142] The terms DNA and RNA as referred to herein mean
deoxyribonucleic acid and ribonucleic acid, respectively. The term
"polynucleotide" means a polymeric form of nucleotides of at least
10 bases in length, either ribonucleotides or deoxynucleotides or a
modified form of either type of nucleotide. The term includes
single and double stranded forms.
[0143] The term "molecular weight" refers to the sum of the atomic
weights of all atoms constituting a molecule, and can be
numerically expressed in Dalton (Da). For example, certain low
molecular weight biologically active compounds may include
compounds having a molecular weight of less than 1,000 Da (1 kDa).
By contrast, certain large molecular weight biologically active
compounds may include drugs having a molecular weight greater than
1,000 Da. In the one embodiment of the invention, large molecular
weight biologically active compounds include compounds having a
molecular weight greater than that of insulin (which has a
molecular weight of about 5,730 Da). In certain embodiments, a
large molecular weight biologically active compound includes
insulin like growth factor-one (IGF-1) with a molecular weight of
about 7,649 Da, human growth hormone (HGH) with a molecular weight
of about 22,124 Da (22 kDa), Botulinum Toxin Type A with a
molecular weight of about 69,000 Da (69 kDa), and various forms of
hyaluronic acid (HA) with molecular weights between 150,000 Da (150
kDa) and 2,180,000 Da (2,180 kDa).
[0144] The term "intact skin" refers to skin that retains its
natural barrier function, and has not been altered by chemical,
physical poration or has not been degraded by prolonged storage or
otherwise altered in a way that may harm the barrier function.
Intact skin has not been subjected to barrier function damage
caused by, but not limited to, freeze/thaw ice crystal damage,
electroporation, laser poration or obliteration, physical poration
by microneedles, enzymatic or chemically induced degradation, or
any other method that may irreversibly degrade normal barrier
function of the skin.
[0145] The term "passive transdermal delivery" refers to drug
delivery wherein a drug is placed on the surface of a skin and
permeates into the skin as a function of concentration gradient
between the higher drug concentration on the skin surface and the
lower drug concentration within the skin. Typical transdermal patch
systems that use passive transdermal drug delivery may include
nicotine patches, estradiol patches and fentanyl patches that are
applied to intact skin and result in the drug permeating into the
skin.
[0146] The term "active transdermal delivery" refers to methods
wherein energy in some forms is imposed on a transdermal system,
resulting in an increase in drug flux into and through the
skin.
[0147] Gelling agents according to this invention are compounds
that can behave as reversible or non-reversible networks. Under
certain conditions, a gelling agent can be placed in a solvent to
form a viscous solution. Under other conditions, that same gelling
agent can be placed in the same or different solvent to form a gel.
The role of gelling agents according to the invention is to prevent
evaporation loss of the biologically active substance in the
appropriate solvent. Examples of gelling agents include, but are
not limited to, hydroxyethyl cellulose, Natrasol.RTM., pectines,
agar, alginic acid and its salts, guar gum, pectin, polyvinyl
alcohol, polyethylene oxide, cellulose and its derivatives,
propylene carbonate, polyethylene glycol, hexylene glycol sodium
carboxymethylcellulose, polyacrylates,
polyoxyethylene-polyoxypropylene block copolymers, pluronics, wood
wax alcohols, tyloxapol, and carbomers.
[0148] The term "photocatalytic agent" refers to any semiconductor
having a wide band gap energy. In an embodiment of the invention,
the band gap energy is on the order of about 2.9-3.2 eV. A band gap
on this order allows infrared and the entire visible spectrum to be
transmitted upon excitation of an electron from the valence band to
the conduction band. Without being bound by theory, pulsed
incoherent light energy that is stored and released from the wide
band gap semiconductor can enhance the bond vibration of a
biologically active molecule also present during this excitation.
The stimulation of active molecules with the transfer of energy
from the semiconductor at discrete wavelengths and pulse rates can
enhance the transport of that molecule across biological membranes,
while the semiconductor can also protect the skin from harmful
ultraviolet (UV) rays by absorbing UV light. By modulating the
wavelength of excitation with that of the band gap energy, the
production of free radicals is avoided entirely. Accordingly, the
use of rutile form of titanium dioxide (TiO.sub.2) as the
photocatalytic agent is preferred because it has a band gap energy
of about 2.9 to 3.0 eV. Other photocatalytic agents suitable for
this invention include, but are not limited to, anatase TiO.sub.2,
brookite TiO.sub.2, ZnO, ZrO.sub.2 and Sc.sub.2O.sub.3. According
to the invention, doped semiconductors can also be used.
[0149] In some embodiments, the "photocatalytic agent" does not
produce a photochemical reduction nor is it intended to produce a
photochemical reaction, but serves as a light scattering agent
within the composition, and acts as a light absorbing/energy
releasing medium to enhance the photokinetic activity. In certain
embodiments, the "photocatalytic agent" herein is absent or not
used in the photokinetic delivery method.
[0150] It should be noted that the "photocatalytic agent" here does
not cause a photochemical reaction, and thus the photokinetic
delivery system herein is not a form of photodynamic therapy. The
photokinetic system of the invention does not cause tissue damage
or chemically alter the drug being transported. For example, the
ELISA analytical methods used for certain drugs being delivered
tend to show that the drug has not been chemically altered, as the
ELISA method would not work on a chemically altered drug (since it
would change the binding property of the drug in the ELISA assay).
This is consistent with Applicants' observations of pharmaceutical
experiments performed in diabetic rats, wherein the blood sugar
levels were reduced with insulin delivered by the photokinetic
method (Kulp et al., Photokinetic Transdermal Drug Delivery
(PTDD)--A Novel Platform Technology for Insulin Delivery: Diabetes
Technology Society Meeting, Nov. 11-13, 2010). In addition,
compounds analyzed by HPLC methods did not show divergent peaks in
the chromographs of the analyte, further demonstrating intact
unaltered/undamaged molecules after delivery by the instant
photokinetic method.
[0151] Additional data shows that the subject system does not
change the tissue morphology as determined by histological
examination (Koutrouvelis et al., Photokinetic Transdermal Delivery
of Cyanocobalamin: Proceedings of The Controlled Release Society,
2008). This feature can be particularly beneficial in satisfying
the safety requirements, when applying the subject method in, for
example, the cosmetic field.
[0152] The term "solvent" according to the invention is any aqueous
or organic solvent that can be combined with the biologically
active agent to form a solution. In one embodiment, the aqueous
solvent is water. In another embodiment, the solvent can be an
aqueous solution of either ethyl lactate or propylene glycol, both
of which act as permeation enhancers. In other embodiments, the
solvent comprises squalene, squalane, and/or olive glycerides or
similar compounds that can assist in dissolving the biologically
active ingredient (such as an cosmetic components). These agents
may also enhance passage of the biologically active ingredient
through the skin layers. Alternately, the term "solvent" can also
mean an adhesive used to embed a biologically active substance, for
example, in a patch. Solvent can also refer to a
pharmaceutically-acceptable medium combined with the biologically
active substance to be used in powder form.
[0153] In another embodiment, the biologically active substance can
be emulsified. For example, lipophilic compounds, such as vitamins
A, D, and E or olive glycerides, can be dispersed in an aqueous
solvent to which an emulsifying agent, such as surfactants or self
emulsifying oils can be added.
[0154] Likewise, in the absence or presence of a solvent, the
biologically active agent according to the invention can also be
combined with a carrier or adjuvant, a substance that, when added
to a therapeutic, speeds or improves its action (The On-Line
Medical Dictionary website: www dot cancerweb dot ncl dot ac dot
uk, under subfolder/omdlindex.html). Examples of adjuvants include,
for example, Freud's adjuvant, ion exchanges, alumina, aluminum
stearate, lecithin, buffer substances, such as phosphates, glycine,
sorbic acid, potassium sorbate, partial glyceride mixtures of
saturated vegetable fatty acids, glycerin, waters, salts or
electrolytes, such as Protamine sulfate, disodium hydrogen
phosphate, sodium chloride, zinc salts, colloidal silica,
magnesium, trisilicate, celluslose-based substances and
polyethylene glycol. Adjuvants for gel base forms may include, for
example, sodium carboxymethylcelluslose, polyacrylates,
polyoxyethylene-polyoxypropylene-block copolymers, polyethylene
glycol and wood wax alcohols.
[0155] Although not required to facilitate transdermal delivery,
skin-penetrating agents, for example, propylene glycol, DMSO, oleic
acid, laurocapram (Azone), cineol, liposomes and nanosomes, can
also be present in the compositions according to the invention.
[0156] The term "donor solution" or "delivery medium" comprises the
biologically active substance itself or any mixture of this
substance with a solvent, a gelling agent, a photocatalytic agent,
a carrier or adjuvant, a skin-penetrating agent, a
membrane-penetrating agent and combinations thereof. The
biologically active substance, or alternately "active ingredient"
does not have to be dissolved in a solvent but can be suspended or
emulsified in a solvent. The donor solution or delivery medium can
take the form of an aqueous or an organic liquid, a cream, a paste,
a powder, a patch, or a mask, such as the tissue or non-tissue type
of mask routinely used in spas, beauty salons, and other cosmetic
installations.
[0157] Although not an exhaustive list, examples illustrating the
term "mammal" include human, ape, monkey, rat, pig, dog, rabbit,
cat, cow, horse, mouse, and goat. Skin surfaces or membranes
according to the invention refer to those of a human or other
mammal.
[0158] The term "viscous solution" refers to a solution that has an
increased resistance to flow.
[0159] The term "cellular surface" refers to an outer layer of the
skin or a cell membrane. Human skin is comprised of three layers:
the epidermis or stratum corneum, the dermis and the hypodermis.
The stratum corneum forms the outermost layer of the epidermis and
consists of about 10 to about 20 layers of flattened, closely
packed cells without nuclei having a thickness of about 10 to about
20 .mu.m. The stratum corneum serves as a barrier to many
substances and is selectively permeable to water and other
compounds. On the other hand, the epidermis, having a thickness of
about 50 to about 100 comprises rapidly dividing basal cells that
flatten as they move into the stratum corneum.
[0160] Finally, the innermost layer of skin, the dermis, comprises
a matrix of various cells including collagen and other fibrous
proteins and has a thickness of about 1 to about 3 mm. It is this
layer that houses hair follicles, sebaceous glands and sweat
glands. The term "transdermal" refers to the penetration and
movement of a biologically active substance through the epidermis
and dermis, or epidermis, dermis and hypodermis.
[0161] The term "transmembrane" refers to the penetration and
movement of a biologically active substance from an extracellular
environment to an intracellular environment.
[0162] The term "intradermal and/or intramembrane deposition"
refers to the deposit of the biologically active substance within
the stratum corneum, epidermis and/or the dermis of skin or within
the various tissue layers or cell structures comprising a
biological membrane. This definition is in contrast to "transdermal
or transmembrane" that may include the passage of biologically
active substances through the tissues into other underlying tissues
thereby translocating beyond the tissues themselves.
[0163] The term "percutaneous penetration" refers to molecules that
have by-passed the dermal blood supply and have diffused into
tissue layers below the dermis. This may be useful for the various
pharmaceutical applications of the subject system and methods.
[0164] The term "incoherent light" refers to electromagnetic waves
that are unorganized and propagate with different phases. The term
"pulsed incoherent light" is any incoherent light having a discrete
ON and OFF period.
[0165] In contrast, "coherent light" refers to all light rays that
are in phase and oriented in the exact same direction to produce a
concentrated beam of light. Lasers generate these types of rays and
can penetrate through materials such as solid media, including
metals (e.g., sheet metal).
[0166] The term "light emitting diode (LED)" is a device that
generally emits incoherent light when an electric voltage is
applied across it. Most LEDs emit monochromatic light at a single
wavelength that is out of phase with each other. According to the
invention, most, if not all, types of LEDs can be used. For
example, an LED having output range from red (approximately 700 nm)
to blue-violet (approximately 350 nm) can be used. Similarly,
infrared-emitting diodes (IRED) which emit infrared (IR) energy at
830 nm or longer can also be used.
[0167] "Optically clear medium" or "light pad" is a material that
acts as a filter to all wavelengths except those wavelengths
emitted from a light source. In a preferred embodiment, the light
pad is comprised of clear poly(methyl methacrylate) or clear
silicon rubber.
[0168] The term "reflective coating or layer" is a material that is
coated on at least one surface of the light pad. Those skilled in
the art will appreciate that the reflective layer can be a
wavelength specific reflective coating (e.g., aluminum, ZnO, silver
or any reflective paint).
[0169] The term "photokinetic" refers to a change in the rate of
motion in response to light, as an increase or (blue part missing
from EPO) decrease in motility with a change in illumination.
[0170] One embodiment of the invention relates to compositions for
photokinetic transdermal and transmembrane delivery of a
biologically active substance using preferably pulsed incoherent
light or, alternatively, regulated coherent light. The composition
may comprise a biologically active substance as the delivery
medium.
[0171] The composition may alternatively comprise a biologically
active substance and a solvent. The percent of biologically active
substance in solvent can be in the range of between 0.0001 to
99.9999% (w/v). Preferably, the biologically active substance is
present in a concentration range of between about 0.01% to about 2%
(w/v). More preferably, the biologically active substance is
present in a concentration range of between about 0.1 mg/ml to
about 10 mg/ml in the solvent or, alternatively, between about
0.01% to about 1% (w/v). Due to the high level of permeation
achieved by the methods and devices described herein, low
concentrations of a biologically active substance in solvent or in
other compositions described herein can be used for efficient
transdermal or transmembrane delivery.
[0172] The composition may instead comprise a biologically active
substance, a gelling agent and a solvent. The percent gelling agent
in a solution of biologically active substance can vary depending
on the type of gelling agent used. For example, Klucel is typically
used at 1% (w/v), Natrasol at 1.5% (w/v), Carbopol at 0.75% (w/v),
and hyaluronic acid is used in a concentration range of 0.25% to 2%
(w/v) for, e.g., high molecular weight HA (and can be adjusted to
even higher percentage for lower molecular weight HA if
necessary).
[0173] Still further, the composition may comprise a biologically
active substance, a photocatalytic agent and a solvent. Preferably,
the photocatalytic agent has a band gap energy of between about 2.9
eV and about 3.2 eV and preferably is present in the composition at
a concentration of between about 0.001% and 20% (w/w). More
preferably, the photocatalytic agent is present in the composition
at a concentration of 2% (w/w).
[0174] Finally, compositions according to the invention may
comprise a biologically active substance, a gelling agent, a
photocatalytic agent and a solvent. Preferably, the photocatalytic
agent has a band gap energy of between about 2.9 eV and about 3.2
eV and preferably is present in the composition at a concentration
of between about 0.001% and 20% (w/w). More preferably, the
photocatalytic agent is present in the composition at a
concentration of 2% (w/w). The biologically active substance
preferably is present in the composition at a concentration of
between about 0.01% and about 2% (w/v). The gelling agent
preferably is present in the composition at a concentration of
between 0.1% and 10% (w/v).
[0175] The biologically active substance of the above compositions
may be selected from the group consisting of chemicals, cosmetics,
drugs, antibiotics, peptides, hormones, proteins, DNA, RNA, various
cosmetic active ingredients, and mixtures thereof.
[0176] The chemical may be a polar or non-polar compound. The polar
compound may be selected from the group consisting of
theophylline-7 acetic acid, sodium ascorbyl phosphate, ascorbic
acid, ascorbyl palmitate, pyridoxine and nicotinic acid. For
example, the polar compound may be pyridoxine. The non-polar
compound may be selected from the group consisting of theobromine,
theophylline, caffeine, and nicotinamide.
[0177] The drug may be selected from the group consisting of
analgesics, anaesthetics, antacids, antianxiety drugs,
antiarrhythmics, antibacterials, antibiotics, anticoagulants and
thrombolytics, anticonvulsants, antidepressants, antidiarrheals,
antiemetics, antifungals, antihistamines, antihypertensives,
anti-inflammatories, antieoplastics, antipsychotics, antipyretics,
antivirals, barbiturates, beta-blockers, bronchodilators, cold
cures, corticosteroids, cough suppressants, cytotoxics,
decongestants, diuretics, expectorants, hormones, hypoglycemics,
immunosuppressives, laxatives, muscle relaxants, sedatives, sex
hormones, sleeping drugs, tranquilizers, and vitamins. In one
embodiment, the anaesthetic is lidocaine.
[0178] The compositions according to the invention may also
comprise antibiotics as the biologically active substance.
Antibiotics according to the invention are selected from the group
consisting of amoxycillin, ampicillin, penicillin, clavulanic acid,
aztreonam, imipenem, streptomycin, gentamicin, vancomycin,
clindamycin, ephalothin, erythromycin, polymyxin, bacitracin,
amphotericin, nystatin, rifampicin, teracycline, coxycycline,
chloramphenicol, and zithromycin. In one embodiment, the antibiotic
is amphotericin B.
[0179] Similarly, in another embodiment of the invention, the
biologically active substance is a peptide selected from the group
consisting of glycine-tyrosine (Gly-Tyr), valine-tyrosine-valine
(Val-Tyr-Val), tyrosine-glycine-glycine-phenylalanine-methionine
(Tyr-Gly-Gly-Phe-Met) (SEQ ID NO: 1),
tyrosine-glycine-glycine-phenylalanine-leucine (Tyr-Gly-Gly-PheLeu)
(SEQ ID NO: 2), and aspartic
acid-arginine-valine-tyrosine-isoleucine-histidine-proline-phenylalanine
(Asp-Arg-Val-TYr-Ile-His-Pro-Phe) (SEQ ID NO: 3).
[0180] The hormone may be selected from the group consisting of
methionine enkephalin acetate, leucine enkephalin, angiotensin II
acetate, .beta.-estradiol, methyl testosterone, progesterone, and
insulin.
[0181] The protein may be selected from the group consisting of
enzymes, non-enzymes, antibodies, and glycoproteins. In one
embodiment of the invention, the protein is an enzyme.
[0182] Numerous biologically active substances useful in the
cosmetic field/industry can also be delivered using the subject
photokinetic intradermal delivery methods, optionally with the
subject dermal patch. One category of cosmetically useful
biologically active substance includes all skin components (such as
collagen, natural or its synthetic forms) that could be readily
replaced, replenished, or supplemented with exogenous molecules
delivered by the subject methods. Merely to illustrate (and thus
not limiting), the following categories of cosmetically beneficial
molecules may be delivered using the subject methods:
antioxidant-photoprotection molecules, such as Vitamin E
(Tocopherols, in particular Alpha-tocopherol, and Tocotrienols),
Ascorbate, Carotenoids (Beta-carotenoid-/Vitamin A, Lycopene,
Zeaxantine, Lutein), Coenzyme Q (Ubiquinone, Idebenone),
Glutathione (including Glutathion derivates: Ethyl ester, Cystine,
etc.), Alpha-glycolic acid, SOD (Superoxide Dismutase), Catalase,
Glutathione peroxydase, Reductase, Taurine, and Alpha-lipoic acid;
polyphenols such as epsilon-viniferin, mixtures of resveratrol and
epsilon-viniferin, and mixtures of polyphenolic, in particular
stilbene and/or falvonol, oligomers and/or polymers; molecules with
healing power, such as Melanin, Glycerol, peptides, or growth
factors; Glycoproteins, such as Sialic acid (moisturizing action,
infective prevention), GAGs (Chondroitin sulphate, Dermatan
sulphate, Cheratan sulphate, Eparin, Eparan sulphate,
Hyaluronates), Decorin; Collagene fibers--other fibers, such as
Hydroxyproline (+Fe, +Vitamin C), Hydroxylysine, Glycine,
Tropocollagen, Reticulin, Keratin, Elastin, MMPs (Matrix
MetalloProteinases); beta-glucans; phytosterols; Anti-aging
molecules, such as Arginine, Citrulline, Ceramides, Carnosine,
Lysine, Inositol, Cysteine, Squalene, Squalane, Chitin, Sericine,
peptides, or growth factors.
[0183] Compositions according to the invention can also contain a
gelling agent in combination with the biologically active agent and
solvent. The gelling agent may be selected from the group
consisting of hydroxyethyl cellulose, hyaluronic acid, Natrasol,
pectines, agar, alginic acid and its salts, guar gum, pectin,
polyvinyl alcohol, polyethylene oxide, cellulose and its
derivatives, propylene carbonate, polyethylene glycol, hexylene
glycol sodium carboxymethylcellulose, polyacrylates,
polyoxyethylene-polyoxypropylene block copolymers, pluronics, wood
wax alcohols, and tyloxapol. In one embodiment, the gelling agent
is hydroxypropyl cellulose. In one embodiment, the gelling agent is
hyaluronic acid.
[0184] Compositions according to the invention can also include a
photocatalytic agent having a wide band gap energy. In one
embodiment, the photocatalytic agent has a wide band gap of between
about 2.9 eV and about 3.2 eV. In a preferred embodiment, the
photocatalytic agent is a rutile form of titanium dioxide
(TiO.sub.2). In another embodiment, the photocatalytic agent is an
anatase form of TiO.sub.2, brookite form of TiO.sub.2, ZnO,
ZrO.sub.2 and Sc.sub.2O.sub.3.
[0185] The composition may also comprise a solvent that is an
aqueous or organic solvent. In one embodiment, the aqueous solvent
is water. In yet another embodiment, the aqueous solvent is an
aqueous solution of ethyl lactate or propylene glycol. The water
may be HPLC grade or purified by means such as reverse osmosis or
distillation.
[0186] The donor solution or delivery medium according to the
invention is comprised of a biologically active substance itself or
any mixture of a biologically active substance with a solvent, a
gelling agent, a photocatalytic agent, a carrier or adjuvant, a
skin-penetrating agent, emulsifier, one or more different
biologically active substances, polymers, excipients, coatings
and/or combinations thereof. In essence, the biologically active
substance or substances can be combined with any combination of
pharmaceutically acceptable components to be delivered to the
cellular surface by the method described herein, e.g., photokinetic
transdermal and transmembrane delivery. The biologically active
substance does not have to be dissolved in a solvent but can be
suspended or emulsified in a solvent. The donor solution or
delivery medium can take the form of an aqueous or an organic
liquid, a cream, a paste, a powder, or a patch. The donor solution
can also comprise microspheres or nanospheres of biologically
active substances.
[0187] The invention described herein is particularly useful for
transdermal delivery of compounds containing chromophores. Without
being bound by any particular theory, it is believed that the
molecular chromophore absorbs photon energy and/or the energy from
an excited photocatalytic agent. As the chromophore returns to
ground state, it generates kinetic energy. With each pulse of
incoherent light, the chromophore's vibration will incrementally
clear a pathway through the skin.
[0188] Consistent with this theory, chemical and heat energy can
cause molecular shape changes and increases in Brownian motion
within a system. These energies cannot be easily cycled and are not
directional. Light energy, on the other hand, is readily defined by
wavelength and can be easily controlled for cycle time stimulation
and incident direction. Molecular conformational changes as a
result of optical stimulation is a widely known phenomenon
exhibited by many classes of compounds with degrees of shape change
determined by the individual molecular structure. The
conformational changes initiated by the exemplary photokinetic
method are reversible; the molecules tend to revert to a resting
state as the optical energy is dissipated during the OFF part of
the cycle. Thus, cyclic light stimulation creates a repeated and
reversible molecule shape change or gross physical movement on a
molecular scale.
[0189] In addition, according to Le Chatelier's principle, if a
system in chemical equilibrium is subjected to a disturbance, it
tends to change in a way that opposes this disturbance. Molecular
systems move in the direction to reduce the external stimuli;
molecules tend to move away from an energy source. Kausar et al
(Photocontrolled translational motion of a microscale solid object
on azobenzene-doped liquid-crystalline films. Angew Chem Int Ed
Engl. 48(12): 2144-79, 121, 2009) have published a paper detailing
how a molecule can be translocated around a surface as a response
to light stimulation, generally by moving away from the light
energy. In the photokinetic system, the drug is applied to the
tissue surface and illuminated from a direction opposite the
tissue. If the molecule is to escape the light energy stimulation,
then it moves in a direction away from the stimulation and into the
tissue. The repeated cycling of the molecular shape adds a gross
movement aspect to the system much like sand going through a
vibrating sieve. The possible interaction of cycled incident
incoherent light on the skin itself may also cycle the tissue
structure and transdermal pathways. These cycled membrane changes
may impart a pumping action on the pathways through the tissue much
like a sieve with cycled pore sizes furthering the sand/sieve
analogy.
[0190] Similarly, the invention described herein is also useful for
transmembrane delivery of biologically active substances. For
example, a person of skill in the art could inject a
therapeutically effective pharmaceutical substance, such as a
chemotherapeutic agent, next to a solid tumor mass. An LED that is
embedded or held next to the tumor mass can be used to deliver the
therapeutic substance from the extracellular environment to the
intracellular environment, effectively causing apoptosis in the
targeted area.
[0191] In addition to compositions, the invention also provides
methods of photokinetic delivery of biologically active substances
using pulsed incoherent light. One method includes preparing a
solution comprising a biologically active substance and a solvent,
applying the solution to a cellular surface, illuminating the
solution on the cellular surface with a pulsed incoherent light
having a selected wavelength, pulse rate and duty cycle and
allowing the solution to permeate the cellular surface. In another
embodiment, the method includes preparing a solution comprising a
biologically active substance, a solvent and a gelling agent,
applying the solution to a cellular surface, illuminating the
solution on the cellular surface with a pulsed incoherent light
having a selected wavelength, pulse rate and duty cycle and
allowing the solution to permeate the cellular surface. In yet
another embodiment, the method includes preparing a solution
comprising a biologically active substance, a solvent, a gelling
agent and a photocatalytic agent, applying the solution to a
cellular surface, illuminating the solution on the cellular surface
with a pulsed incoherent light having a selected wavelength, pulse
rate and duty cycle and allowing the solution to permeate the
cellular surface. In a preferred embodiment, the cellular surface
is an outer layer of a skin of a mammal or a cell membrane.
3. Illustrative Photokinetic Devices
[0192] FIG. 1 illustrates testing device 100 in accordance with the
invention. Testing device 100 provides photokinetic transdermal and
transmembrane delivery of biologically active substances to a
portion of skin or membrane by illuminating the biologically active
substance with pulsed incoherent light. Testing device 100 includes
a light source 3 that illuminates a biologically active substance
in donor cell 4 such that the biologically active substance
diffuses into skin 6 with little to no damage to skin 6. Testing
device 100 can also be arranged such that the light source 3
illuminating a biologically active substance in donor cell 4 is
parallel to a surface on which it is mounted.
[0193] Testing device 100 preferably includes a driver circuit 2
that provides control signals to light source 3 such that pulsed
incoherent light is provided to donor cell 4. Driver circuit 2 may
also provide control signals that control the intensity, direction,
and/or frequency of light source 3. A pulsed incoherent light may
advantageously reduce damage to skin 6 as compared to a continuous
light source, and provides photokinetic transdermal and
transmembrane delivery of biologically active substances within
donor cell 4 to skin 6.
[0194] Driver circuit 2 may regulate an electrical signal that
turns (i.e., switches) light source 3 ON and OFF at a particular
frequency. Such an electrical signal may be provided, for example,
by a voltage generator. Alternatively, driver circuit 2 may itself
be a voltage generator and may produce an electrical signal to
control the switching characteristics of light source 3. For
example, a voltage generator coupled to light source 3 may provide
a square wave to power light source 3. This square wave may have a
desired period such that light source 3 provides incoherent light
with a desired frequency (e.g., a square wave period of 0.5 seconds
would cause light source 3 to switch at 2 Hz).
[0195] Light source 3 preferably provides incoherent light (to
avoid any potential damage done to skin 6 during the use of testing
device 100). Light source 3 may be, for example, an LED, halogen
light source, fluorescent light source, natural light, or other
source of light. More particularly, light source 3 can be a light
emitting diode (LED) (fluorescence, 350-1700 nm) or an infrared
light emitting diode (ILED) or a Mercury-Argon (253-922 nm), pulsed
xenon (UV-VIS, 200-1000 nm), deuterium (UV, 200-400 nm),
deuterium/halogen (UV/VIS/NIR, 200-1700 nm) or tungsten halogen
(color/VIS/NIR, 360-1700 nm) light source. Light source 3
preferably is operable in the range from red (approximately 700 nm)
to blue-violet (approximately 350 nm). Similarly, infrared-emitting
diodes (IREDs) that emit infrared energy at 830 nm or longer may be
used.
[0196] Light source 3 does not have to be an incoherent light
source. Alternatively, light source 3 may be a coherent light
source such as, for example, a laser. In that case, driver circuit
2, or other regulation circuitry, is preferably used to turn a
coherent light source 3 ON and OFF to reduce the amount of damage
to skin 6 while still photokinetically delivering a biologically
active substance to donor cell 4. Furthermore, a light
regulation/conversion device may be placed between a coherent light
source 3 and donor cell 4 to convert the coherent light to
incoherent light.
[0197] Note that a device such as driver circuit 2 or a controlled
voltage generator is not required to pulse light source 3.
Alternatively, shutter 11 may be employed between light source 3
and donor cell 4. Such a shutter selectively OPENs and CLOSEs such
that donor cell 4 is supplied pulsed incoherent light from light
source 3. The speed at which the shutter OPENs and CLOSEs
determines the frequency of the light pulsed onto the skin. Filters
(not shown) may also be placed between light source 3 and donor
cell 4 in order to remove, for example, light of specific
wavelengths that may damage skin 6. Alternatively, light source 3
may be immersed in a solution found in donor cell 4. Preferably,
the wavelength of light reaching skin 6 is chosen not only to
reduce damage to skin 6, but also to increase the photokinetic
activity in donor cell 4 (e.g., 350 nm to 450 nm). The pulse rate
of such light may also be between 1.7 cps and 120 cps (e.g., 24
cps). If fluorescent light is employed as light source 3, it
preferably has a wavelength range from about 260 nm to about 760
nm. If ultraviolet, visible, near infrared, or halogen light is
employed as light source 3, the light source preferably has a
wavelength range from about 340 nm to about 900 nm. The invention,
however, is not limited to the these wavelengths.
[0198] Donor cell 4 holds a biologically active substance (e.g.,
chemicals, drugs, antibiotics, peptides, hormones, proteins, DNA,
RNA and mixtures thereof). Donor cell 4 may also include a solvent
that forms a solution with the biologically active substance. The
solution may also include a photocatalytic (having, for example, a
band gap energy of between about 2.9 eV and about 3.2 eV) and/or a
gelling agent. The solvent may be an aqueous or an organic solvent.
Furthermore, skin 6 may be a cellular surface which is an outer
layer of a skin. Generally, skin 6 may be any medium that allows at
least the biologically active portion of donor cell 4 to diffuse
into that medium in response to that medium being exposed to light
source 3. In one embodiment, this medium is a cell membrane for
transmembrane delivery.
[0199] Clamp 5 is preferably included in testing device 100 to
couple donor cell 4 and skin 6 to receiving cell 7. Receiving cell
7 may be present in container 17 as a result of diffusion of at
least the biologically active portion of donor cell 4 through skin
6. Also, receiving cell 7 may contain a solvent, e.g., HPLC grade
water, wherein diffusion of at least the biologically active
portion of donor cell 4 through skin 6 enters into the solvent.
Generally, the concentration of the biologically active substance
is higher in donor cell 4 than in receiving cell 7. Skin supports
16 may also be included in order to position skin 6 above container
17 and below light source 3. Donor cell 4 is located in container
14 and preferably contacts an area of skin 6. Container 14 and
container 17 may be the same container. Furthermore, a skin
aperture (not shown) may exist to receive at least a portion of
skin 6 such that skin 6 separates container 14 from container
17.
[0200] Temperature control device 8 is preferably applied to at
least a portion of container 7. Temperature directors 18 may be
included as a part of container 17 or coupled to container 17 to
direct temperature control device 8. Temperature directors 18 may
also be used to structurally provide support for a heat source such
as a heat bath. For example, hot water may be placed in housing
defined by temperature directors 18 and a portion of container 17
between temperature directors 18. Further to this example, a heat
source may be used to heat such water. Alternatively, a heat source
may be directly coupled to container 17. Preferably, temperature
control device 8 heats container 17 to a constant level. While the
temperature of the solvent in receiving cell 7 can vary, it is
preferably about 37.degree. C., human body temperature, or about
33.5.degree. C., human skin surface temperature. For applications
requiring container 17 to be cooled, temperature control device 8
may additionally or alternatively be a cooling source. A
temperature sensor (not shown) may be placed in, on, or about
container 17 or a heat source such that temperature control device
8 keeps container 17 at a particular temperature for a particular
period of time.
[0201] Stir bar 9 may be included in container 17 to stir any
solution in container 17. Preferably, stir bar 9 constantly stirs
the solution in container 17. Container 17 may be alternatively
stirred, for example, by a shaking device. Removal of stir bar 9
would, for example, allow container 17 to be easily sanitized while
reducing the design complexity of container 17. Stir bar 9 may be
connected to an electrical motor (not shown).
[0202] Port 10 may be included in container 17 to add or remove
samples to or from receiving cell 7 or solutions to or from
container 17. Generally, port 10 is an aperture in container 17.
Guide tube 12 may form an extended port 10 such that a sample
recovery or dispersal tool can easily migrate to port 10. Cover 13
may be employed on port 10 (or guide tube 12) such that
contaminants from outside container 17 do not pass through port 10
when samples are being added or removed from container 17. Guide
tube 12 is generally an adapter. For example, if the
recovery/dispersal tool is a needle, then guide tube 10 preferably
facilitates the coupling of the needle to port 10.
[0203] Lens 23 may be included in testing device 100 to, for
example, focus light source 3 on donor cell 4 or to provide a
transparent medium in which light from light source 3 may pass onto
donor cell 4 while contaminants from outside container 14 are
isolated from donor cell 4. Lens 23 may be a transparent medium,
such as, for example, a transparent polymer or glass.
[0204] Container 17 may include insulation 15 to control the amount
of heat supplied to container 17. Insulation 15 may also be part of
a heat bath and may be filled with water. The amount of insulation
15 about temperature control device 8 may be reduced such that
temperature control device 8 affects the temperature of container
17 more than ambient heat.
[0205] FIG. 2 illustrates testing device 200 in accordance with the
invention. Testing device 200 includes light pad 201 and is
otherwise similar/identical to testing device 100 of FIG. 1. Light
pad 201 includes at least one and preferably more than one light
source 3, which is preferably an LED. Light pad 201 is preferably
fabricated from an optically clear material (e.g., poly (methyl
methacrylate) or silicone rubber). Similar to testing device 100,
testing device 200 can also be oriented differently than shown.
[0206] FIG. 3A illustrates light pad 300 in accordance with the
invention. Light pad 300 includes driver circuit 302, base 312,
light source 314, and wiring 315. Wiring 315 may be included to
electrically couple control device 302 (or a power supply) to one
or more light sources 314, and wiring 315 may have a protective
sheath. Base 312 is preferably a silicon substrate in which light
sources 314 are fabricated. Light sources 314 are preferably
incoherent sources of light and are preferably LEDs having a narrow
bandwidth. Alternatively, other types of light sources may be used.
Light sources 314 may be turned ON and OFF by driver circuit 302
either as a group, individually, or in sections. For example, light
sources 314 may be arranged as multiple arrays of light sources.
Driver circuit 302 may then selectively pulse only a single array
of light sources 314 such that only a desired portion of a medium
(e.g., skin 6 from FIGS. 1 and 2) receives pulsed light. Moreover,
multiple arrays can be included on light pad 300 in which each
array includes LEDs of a specific wavelength. Thus, when only a
specific wavelength is desired or needed, driver circuit 302 can
selectively turn ON the array comprised of LEDs having that
particular wavelength. For example, light pad 300 may include an
array of ILEDs and an array of LEDs where driver circuit 302
selectively switches between the ILED array and the LED array. This
may be desirable when a biologically active substance is more
reactive to or less degraded/denatured by light of a particular
wavelength.
[0207] Instead of having arrays of particular wavelengths, other
characteristics may be utilized. For example, two arrays may have
LEDs of the same wavelength (or different wavelengths), and the
arrays may be of different intensities or may focus light in
different directions. Light sources 314 may be mounted on gears
(not shown) that can be turned/rotated by motors (not shown) and
controlled by driver circuit 302 such that the direction and
intensity of light being provided to a particular area can be
manipulated. Driver circuit 302 may be controlled by computer 325
either directly or via a graphical user interface (GUI).
[0208] Light pad 300 can be, for example, a tanning bed. If light
pad 300 provided coherent light, a light scatter device, filter, or
conversion device can be provided to convert the coherent light
into incoherent light.
[0209] FIG. 3B illustrates light array 313 mounted in base 312.
Array 313 includes two or more light sources 314 electrically
connected in series by wiring 315. If light sources 314 are to
provide light below base 312, reflective layer 316 may be included
above base 312 to reflect light scattered from base material or
skin while base 312 remains a transparent medium. Multiple light
sources 314 may have different wavelengths such that light sources
314 having a particular wavelength may be selectively turned ON and
OFF to provide light of a single selected wavelength or multiple
selected wavelengths. Light sources 314 may provide light above
base 312. In this case, reflective layer 316 may be placed on base
312 (which does not have to be transparent) and beneath light
sources 314 to reflect light above base 312. The reflective layer
can be a wavelength specific reflective coating (e.g., aluminum,
ZnO, silver or any reflective paint).
[0210] The in vitro and in vivo methods and devices of the
invention can also be used in combination with other active
delivery techniques such as ionotophoresis, sonophoresis,
phonophoresis, or used in conjunction with microporation methods
such as microneedles, electroporation, phonophoresis, laser
poration/obliteration.
[0211] A more detailed description of an exemplary embodiment of
the photokinetic device--a Franz cell apparatus, is provided
below.
[0212] Franz Permeation Cell In Vitro Method
[0213] A traditional Franz cell apparatus (PermeGear, Inc,
Bethlehem Pa.) was modified to allow placement of the LEDs into the
donor chamber. The tissue area exposed to drug was 1 cm.sup.2. The
cells were placed in an aluminum block equipped with magnetic
stirrer and maintained at constant temperature. Target tissue
(intact human split thickness skin) was placed between the donor
and recipient chamber and clamped together to create a seal. Drugs
were formulated in appropriate carriers and placed in the donor
chamber. In the recipient chamber, either HPLC grade water or
buffer was used for ease of analysis. Samples from the recipient
chamber fluid are taken through the side port at various time
points. Control "passive permeation" cells were set up the same
way, without the LEDs.
[0214] LEDs of discrete wavelengths were purchased from Roithner
Lasertechnick GmbH, Vienna, Austria, but other equivalent LEDs may
also be suitable. LEDs used are specified as peak emitting
wavelength value in nanometers (nm).+-.spectrum 1/2 width in
nanometers at 50% relative radiant intensity output along with
specified radiated output power in milliwatts (mW) and
manufacturers product number: 350 nm.+-.5 nm 200 .mu.W (RLT350-30),
370 nm.+-.5 nm 1.2 mW (RCL-370-10), 390 nm.+-.5 nm 8.1 mW
(LC503MUV1), 405 nm.+-.5 nm 10 mW (LED405-03V), 436 nm.+-.10 nm 12
mW (LED436-03), and 450 nm.+-.10 nm 20 mW (LED450-06).
[0215] An adjustable pulse rate square wave signal generator
constructed by the inventors providing an equal time ON and OFF
cycle (or 50% duty cycle) was used to drive LEDs. Driver output
pulse frequency is specified as the number of complete ON-OFF
cycles per second (cps), i.e., 24 complete ON-OFF cycles in one
second is specified as 24 cps. The LEDs were driven at or below
manufacturer's specified current by incorporating current limiting
resistors in the drive circuits. Current limiting resistors were
selected based on the formula: resistor value in ohms=(pulse
generator supply voltage-LED specified voltage)/LED specified
current rating (in amperes). Resistors selected were equal to or
greater than the formula determined value in order to prevent
excessive radiant heat generation from the LEDs.
[0216] The subject tissue is placed between the donor chambers and
recipient chambers and held together by a clamp (not shown)
providing sealed separation between the two chambers. The recipient
chamber is filled with distilled water through the sidearm sampling
port. The donor chamber holds the subject drug dissolved in an
acceptable drug carrier formulation. At various time points,
samples are dawn from the side arm port for chemical analysis.
Control cells are set up with the same conditions as the
photokinetic cells, but without the LED.
[0217] The photokinetic Franz cell testing portion includes an
electrical driver circuit (not shown) that provides control signals
to the LED light source. The electronic driver circuit regulates an
electrical signal that turns (i.e., switches) light source ON and
OFF at a particular selected frequency. The electrical drive
current is limited to proved current levels at or below the LED's
specified current to eliminate exogenous heat generation from the
LED. The voltage generator coupled to light source provides an
electrical square wave to power the light source. This square wave
has a desired ON and OFF period such that light source provides
pulsed incoherent light from the LED source with a desired pulse
frequency (e.g., a square wave 50% duty cycle period of 0.5 seconds
ON and a 0.5 seconds OFF would cause light source to switch at 1 Hz
or 1 cycle per second (CPS)). Pulsed light frequency used herein
range from 24 to 100 CPS as reported. Useful ranges of pulse rate
may exceed those reported herein. The preferred embodiment has a
range of 1 to 120 CPS.
[0218] Furthermore, the proportion of ON time vs. OFF time 50% duty
cycle or equal ON:OFF time was determined out of convenience for
the electronic control construction and is not intended to limit
the specifications of the invention. In general, the excitation
time required for molecular stimulation can be quite short, while
the time required to dissipate this energy may be longer. A shorter
period of ON time vs. OFF time, for example an ON time of 10% with
an OFF time of 90% of the total cycle time, may also be useful for
the invention. This may impact the pulse rate frequency
determination, which may then affect the attainable flux rate. An
added advantage of shorter ON time (lower percentage duty cycles)
is that in a battery operated system, this would conserve energy.
Thus in certain embodiments, there is a period ON and OFF
illumination times to provide a cyclical period of light
stimulation with a period of OFF resting time. A discrete ON time
and a discreet OFF time, in the range of for example 1% ON/99% OFF
to 99% ON to 1% OFF (including any integer values in between, such
as 2% ON98% OFF, 5% ON95% OFF, 10% ON90% OFF, 15% ON85% OFF, etc.)
are all contemplated embodiments of the invention, with 50% ON50%
OFF being preferred in certain situations.
[0219] FIG. 4 illustrates an exemplary Franz diffusion cell
apparatus for in vitro determination of photokinetic conditions of
light wavelength and pulse rate, in accordance with the present
disclosure. Franz diffusion cells 31 are shown within a heat block
25, one (B) being shown in partial cut-away for purposes of
clarity. Skin tissues 23 are inspected under six times
magnification for holes or tears or other imperfections. Skin 23 is
placed and floated onto the donor chamber containing recipient
fluid in a method to prevent air bubbles between the recipient
fluid and the skin. The donor chamber 21 is then affixed and held
in place by a clamp (not shown) so that the skin separates the two
Franz cell chambers, donor chamber 21 and recipient chamber 22.
Skin placement is oriented to mimic an in vivo situation with the
dermal skin surface facing the recipient chamber 22 and the
epidermal/stratum corneum surface facing the donor chamber 21.
[0220] The testing device illustrated in FIG. 4, in accordance with
the present disclosure, provides photokinetic transdermal and
intradermal delivery of biologically active substances to a portion
of skin by illuminating a biologically active substance with pulsed
incoherent light. Testing device can include a light source (not
shown) that illuminates a biologically active substance in donor
chamber 21 such that the biologically active substance diffuses
into the skin tissue 23 without damage to the skin tissue 23.
Testing device can also be arranged such that the light source
illuminating a biologically active substance in donor chamber 21 is
horizontal or parallel to a surface on which it is mounted.
[0221] Testing device may include an electrical driver circuit that
provides control signals to the light source such that pulsed
incoherent light is provided to the donor chamber 21. The driver
circuit may also provide control signals that control the
intensity, direction, and/or frequency of the light source. A
pulsed incoherent light advantageously and cyclically illuminates
the subject drug formulation 24 producing a period of excitation
and relaxation of the drug 24 and the skin tissue 23 which provides
photokinetic transdermal translocation of biologically active
substances within donor chamber 21 into and through the skin tissue
23.
[0222] Electronic Driver circuit may regulate an electrical signal
that turns (i.e., switches) light source ON and OFF at a particular
frequency. Such an electrical signal may be provided, for example,
by a voltage generator controlled by an electronic flasher circuit.
Alternatively, a driver circuit may itself be a voltage generator
and may produce an electrical signal to control the switching
characteristics of light source. For example, a voltage generator
coupled to light source may provide an electrical square wave to
power the light source. This square wave may have a desired ON and
OFF period such that light source provides pulsed incoherent light
with a desired pulse frequency (e.g., a square wave period of 0.5
seconds ON and a 0.5 seconds OFF would cause light source to switch
at 1 Hz or 1 cycle per second (CPS)).
[0223] The light source preferably provides incoherent light (to
avoid any potential damage done to skin 23 or cause damage to the
drug or cosmetic active ingredient 24 during the use of testing
device). The light source may be, for example, an LED, halogen
light source, fluorescent light source, natural light, or other
source of light. More particularly, the light source can be a light
emitting diode (LED) (fluorescence, 350-1700 nm) or an infrared
light emitting diode (ILED) or a Mercury-Argon (253-922 nm), pulsed
xenon (UV-VIS, 200-1000 nm), deuterium (UV, 200-400 nm),
deuterium/halogen (UV/VIS/NIR, 200-1700 nm) or tungsten halogen
(color/VIS/NIR, 360-1700 nm) light source. The light source
preferably is operable in the range from red (approximately 700 nm)
to blue-violet (approximately 350 nm). Similarly, infrared-emitting
diodes (IREDs) that emit infrared energy at 830 nm or longer may be
used.
[0224] The light source does not have to be an incoherent light
source. In accordance with aspects of the present disclosure, the
light source may be a coherent light source such as, for example, a
laser. In that case, the driver circuit, or other regulation
circuitry, is preferably used to turn the coherent light source ON
and OFF to reduce the amount of damage to skin tissue 23 while
still photokinetically delivering a biologically active substance
24 from the donor cell 21 into the eye tissue 23. Furthermore, a
light regulation/conversion device may be placed between a coherent
light source in the donor cell 21 to convert the coherent light to
incoherent light.
[0225] Note that a device such as an electronic driver circuit or a
controlled voltage generator is not required to pulse light source.
Alternatively, a mechanical shutter may be employed between light
source and donor cell. Such a shutter selectively OPENs and CLOSEs
such that donor cell is supplied pulsed incoherent light from light
source. The speed at which the shutter OPENs and CLOSEs determines
the frequency of the light pulsed onto the eye tissue. Filters (not
shown) may also be placed between light source and the donor cell
in order to remove, for example, light of specific wavelengths that
may damage the skin or reduce photokinetic activity. Alternatively,
the light source may not need be immersed or optically coupled with
the drug solution found in donor cell. The essential arrangement is
when a drug in contact with the subject's skin is positioned to
receive pulsed incoherent light from a selected source at a
selected pulse frequency.
[0226] The wavelength of light reaching eye tissue may be chosen
not only to reduce damage to the tissue, but also to increase the
photokinetic activity in donor cell (e.g., 350 nm to 450 nm). The
pulse rate of such light may also be between 1.7 cycles per second
(cps) and 120 cps (e.g., 24 cps). If fluorescent light is employed
as light source, it may have a wavelength range from about 260 nm
to about 760 nm. If ultraviolet, visible, near infrared, or halogen
light is employed as light source, the light source may hasve a
wavelength range from about 340 nm to about 900 nm. The invention,
however, is not limited to these wavelengths. Any method to pulse
illuminate the drug that is in contact with the eye tissue may
provide the photokinetic transdermal drug delivery.
[0227] Donor chamber 21 holds a biologically active substance
(e.g., chemicals, drugs, antibiotics, peptides, hormones, proteins,
DNA, RNA and mixtures thereof). Donor chamber 21 may also include a
solvent that forms a solution with the biologically active
substance. The solution may also include a gelling agent, as
appropriate. The solvent may be an aqueous or an organic solvent.
Generally, skin tissue 23 may be any medium that allows at least
the biologically active portion drug formulation 24 contained in
the donor chamber 21 to diffuse into that medium in response to
that medium being exposed to a selected light source pulsed at a
selected pulse rate. In one embodiment, this medium is a skin for
transdermal delivery. In another embodiment the medium is skin
tissue for intradermal delivery.
[0228] A clamp (not shown) may optionally be included in testing
device to couple donor chamber 21 and skin tissue 23 to the
recipient chamber 22. Drug components comprising the donor
formulation 24 placed in donor chamber 21 may be present in
recipient chamber 22 as a result of the diffusion of at least the
biologically active portion 24 of donor chamber 21 through eye
tissue 23. Also, recipient chamber 22 may contain a solvent, e.g.,
HPLC grade water, wherein diffusion of at least the biologically
active portion 24 of donor cell 21 through eye tissue 23 enters
into the solvent. Generally, the concentration of the biologically
active substance is higher in donor chamber 21 than in recipient
chamber 22.
[0229] Temperature control device 25, such as a heat block, is
preferably applied to at least a portion of the recipient chamber
22. Temperature directors may be included as a part of heat block
25 or coupled to the recipient chamber 22 to direct temperature
control device 25. Temperature directors (not shown) may also be
used to structurally provide support for a heat source such as a
heat bath. For example, warm water may be placed in housing defined
by temperature directors and a portion of recipient chamber 22
between temperature directors. Further to this example, a heat
source may be used to heat such water. Alternatively, a heat source
may be directly coupled to recipient chamber 22. Preferably,
temperature control device 25 heats the Franz cell assembly 31 to a
constant level. While the temperature of the solvent in recipient
chamber 22 can vary, it is preferably about 37.degree. C., human
body temperature, or about 35.5.degree. C., human skin surface
temperature. For applications requiring Franz cell assembly 31 to
be cooled, temperature control device 25 may additionally or
alternatively be a cooling source. A temperature sensor (not shown)
may be placed in, on, or about the Franz cell 31 or a heat source
such that temperature control device 25 keeps the Franz cell 31 at
a particular temperature for a particular period of time.
[0230] With continued reference to FIG. 4, stir bar 26 may be
included in recipient chamber 22 to stir any solution in recipient
chamber 22. Preferably, stir bar 26 constantly stirs the solution
in recipient chamber 22. Recipient chamber 22 may be alternatively
stirred, for example, by a shaking device. Removal of stir bar 26
would, for example, recipient chamber 22 to be easily sanitized
while reducing the design complexity of recipient chamber 22
assembly. Stir bar 26 may be connected to an electrical motor (not
shown).
[0231] Side arm port 27 may be included in recipient chamber 22 to
remove samples from and to replace sample volume into recipient
chamber 22 or solutions to or from recipient chamber 22. Generally,
port 27 is an aperture into recipient chamber 22. An alternate
guide tube (not shown) may be included to form an extended port 27
such that a sample recovery or dispersal tool can easily migrate to
port 27. A cover may be employed on port 27 such that contaminants
from outside recipient chamber 22 do not pass through port 27 when
samples are being added or removed from recipient chamber 22. In
accordance with certain aspects of the present disclosure, if a
guide tube is included in association with port 27, the guide tube
is generally an adapter. For example, if the recovery/dispersal
tool is a needle, then guide tube 27 preferably facilitates the
coupling of the needle to port 27.
[0232] The Franz cell apparatuses 31 are designated to determine
passive permeation A or photokinetic permeation B into and through
skin tissues 23. The Franz cell has two chambers--the donor chamber
21 and the recipient chamber 22. Skin tissue 23 is placed between
the two chambers and sealed into place and held between the two
chambers by a clamp (not shown). The recipient chamber 22 is filled
with an aqueous solution selected to allow for chemical analytical
methods. The donor chamber 21 is filled with a drug in a
pharmacologically acceptable formulation, as described herein. The
recipient chamber 22 is constantly stirred by a magnetic stir bar
26. A portion of the recipient chamber is placed in a heat block 25
heated to a physiological temperature (about 35.5.degree. C.). At
various time points, samples are dawn from the side arm port 27 for
purposes of chemical analysis.
[0233] The passive permeation cell A provides permeation flux rates
though the skin tissue. In the photokinetic Franz cell B, a
selected LED 28 is partially submerged within the drug formulation
24 within the donor chamber 21. The LED is driven by an external
pulse generator at a selected pulse rate and connected to the LED
electrical connectors 29. The skin tissue 23 is positioned in
contact with and under the drug formulation 24. The drug
formulation in contact with the tissue is illuminated by the light
30 generated by the LED 28.
[0234] By sampling the fluid from the recipient cells 22 and
performing chemical analysis, comparisons of the two permeation
conditions: A passive transdermal permeation and B photokinetic
transdermal permeation can be determined.
[0235] Now turning to FIG. 5. For drugs to permeate through skin as
in transdermal drug delivery, they must first be transported into
the skin and provide an intradermal presence before diffusing out
of the skin into deeper tissue regions. The intradermal drug
concentration is of particular importance for applications
requiring the biological active substance to reside within the skin
to provide a biological affect on the skin itself. Photokinetic
intradermal/transdermal delivery is an active delivery technology
wherein a biologically active substance permeates into and/or
through skin at a higher flux rate than what could be achieved with
passive diffusion alone. Furthermore, the delivery of the
biologically active substance can be controlled or modulated in
order to allow the concentration of the biologically active
substance to be regulated, and to preferentially acculmulate
intradermally (preferably not penetrating any further) within the
skin and have an effect primarily on the skin itself. FIG. 5
represents the exemplary method to determine drug concentration
within the skin.
[0236] Passive and photokinetic permeation conditions are similar
except for the photokinetic donor chamber is provided with pulsed
incoherent light as pervious described. The method of determining
intradermal concentration of the drug may comprise the following
steps: the donor drug formulation 24 is removed from the donor
chamber by several washes of the donor chamber 21 with demonized
water. Skin is separated from both Franz apparatus chambers 21 and
22. The area of the skin held between the two chambers is cut away
from the skin that was positioned under the flanges of the donor
and recipient chambers providing a section of skin that was exposed
to the experimental conditions. The excised skin sample is then
frozen in liquid nitrogen. The frozen skin sample is then placed in
a cryogenic tissue pulverizer (Fisher Scientific) pre-cooled in
liquid nitrogen and freeze fractured. The pulverized freeze
fractured tissue is weighed and placed into an extraction buffer
appropriate for the selected drug analysis. The tissues are toughly
mixed with the buffer solution and centrifuged to separate the
solid tissue components from the extraction buffer. Supernates from
the centrifuged samples are analyzed for subject drug concentration
and protein concentration. Protein concentration determination is
used to normalize the drug extraction efficiency between the
samples.
[0237] Procurement of Human Skin for in vitro Testing
[0238] Human split thickness skin harvested from cadaver donors
(skin removed with a dermatome including the entire epidermis and
part of the dermis) was obtained from Allosource International
(Centennial, Colo.). Spit thickness skin had a thickness of
approximately 0.625 mm to 2.0 mm thick. Procured skin was placed
immediately in pre-cooled RPMI 1640 medium with
antibiotics/antimycotics and refrigerated at 4.degree. C. until
used. Skin was used within 96 hours after procurement.
4. Patch for Dermal Treatment
[0239] In certain embodiments, the methods of the invention utilize
a dermal patch to facilitate the delivery of a biologically active
substance/molecule. In certain embodiments, the patch may take the
form of a mask for topical use (e.g., face mask). In certain
embodiments, the patch allows the controlled release of active
ingredients therein (e.g., the biologically active
substance/molecule). The dermal patch may in some instance overcome
the hydrophobic resistance typical of the skin.
[0240] In certain embodiments, the subject dermal patch is
biodegradable, and may be of vegetal origin.
[0241] In certain embodiments, the subject dermal patch is a
pre-shaped dermal treatment patch or mask that does not contain
substances like Tissue Non-Tissue (TNT), or polymers such as
polyester or polyacrylate, in which the desired active ingredients
can be inserted.
[0242] In certain embodiments, the subject dermal patch or mask
does not require an adhesive layer in order to be applied to the
skin.
[0243] In certain embodiments, the subject dermal mask or patch is
free of preservatives and/or perfumes.
[0244] In certain embodiments, the subject dermal patch or mask
contains a polysaccharide-based vegetal matrix, which may be
extracted from red algae (e.g., Chondrus crispus), and which may be
rich in polymeric thickeners.
[0245] In certain embodiments, the subject patch or mask are for
use in cosmetic and/or pharmaceutical/therapeutic applications.
[0246] In certain embodiments, the base of the subject patch or
mask is a matrix of 100% plant origin, such as one obtained from an
extract of red algae (e.g., Chondrus crispus), which may contain
polysaccharides with thickening properties, and which may have the
peculiarity to form a biological network. Such polysaccharides are
used in food and cosmetic products for their structuring
properties, e.g. for enhancing viscosity and/or stability.
[0247] The polysaccharides of Chondrus crispus may generate a
network that imparts a three-dimensional structure to the dermal
patch or mask. This structure is similar to that obtained, e.g., by
using silicones, and may confer similar structural properties such
as transparency, elasticity, and flexibility optimal to be easily
applied to the skin; resistance, adhesive strength, possibility to
be removed and replaced while maintaining the same shape, etc.
After use, the patch can be easily removed from the skin without
leaving residues.
[0248] The patch or mask of this invention is a highly viscose
fluid, structured in the form of a gel, which is resistant and can
be safely applied to the human skin. The patch forms a film on the
skin, allowing hydration by water release from its vegetal matrix
as well as gradual and controlled release of any cosmetic,
dermatological, or pharmaceutical active ingredient incorporated in
the formulation.
[0249] These features are maintained as long as the water content
of this structure remains unchanged: as the water evaporates, the
structure tends to dry up and lose the above characteristics.
However, the process of drying is much slower than with other
cosmetic treatments of skin, usually from 10 to 30 min., occurring
only several hours after air exposure.
[0250] Importantly, unlike the commonly used patches, which contain
a synthetic polymeric support to guarantee the structure and
mechanical characteristics of the patch, the matrix of the patch
subject of this invention may be totally natural and thus
biodegradable.
[0251] At lower concentrations of red algae (e.g., Chondrus
crispus) extract, the required structural characteristics might not
be obtained: the product forms a classical viscous gel, but remains
liquid, and does not maintain the shape when subjected to
mechanical stress, such as the removal from the blister or when
applied on the skin.
[0252] Conversely, at higher concentrations of red algae extract,
the mixture becomes clearer and fails to gel.
[0253] At specific concentrations, it is possible to obtain a gel
structure with such interesting mechanical performance.
[0254] The stability of the dermal patch in its original sealed
blister has been tested at 1, 3, and 12 months, by visual
inspection and tests of adhesion to the skin, and no significant
loss of property was observed.
[0255] The patch compositions of the invention can be prepared by
conveniently mixing the components of the mixture at suitable
temperatures.
[0256] The blend of ingredients at the selected temperature is a
viscose fluid that can be mixed with normal equipment, such as
classical mixers.
[0257] The mixture may be kept at constant temperature, from 30 to
90.degree. C., e.g., closer to the upper limit of 90.degree. C.,
under constant stirring at 1 to 30 rpm, e.g., around 10 rpm.
[0258] When cooled down gradually to room temperature, the obtained
product is a highly resistant, transparent solid, showing high
elasticity and adhesion to the skin. The appearance, texture and
resistance of the finished product is comparable to those of
similar silicone-based products, but with the advantage of being
natural.
[0259] As already mentioned above, the product maintains these
structural characteristics until the level of hydration is
constant.
[0260] The product may contain any cosmetic or dermopharmaceutical
active ingredient, incorporated in the solution or dispersed in the
water entrapped by the network of polysaccharides.
[0261] The moulding may take place in a container--a "blister"--of
the desired shape. As the liquid cools, the product takes the shape
in the blister.
[0262] Accordingly, single patches of various size and shape can be
prepared, suited to be used for different applications.
[0263] The exact amount of product can be dosaged through a timed
and heated electronic valve under controlled temperature,
specifically designed and adapted for this process. The correct
opening time of the valve may determine the flow of the product and
the right dosage. The blister containing the product is then
sealed, e.g., by an aluminium/polyethylene tie layer. Then the
blister is die-cut to obtain the desired shape.
[0264] The sealed containers allow the patch to retain the
structural and mechanical properties until the moment of
application.
[0265] In one embodiment of the invention, the initial mixture of
ingredients is prepared by dispersing Chondrus crispus (SETALG) in
a blend of glycol, glycerin, or other moisturising substances until
a homogeneous dispersion is obtained without any lumps.
[0266] In another embodiment, during the initial mixing phase, 75%
of the water needed for the production of the batch is heated to
90.degree. C. After reaching this temperature, the suspension is
added under vigorous stirring, and then it is left to cool to
50-60.degree. C. In this embodiment, the water soluble active
ingredient is dispersed in the remaining water, and then it is
added to the hot phase under stirring to obtain a complete
dispersion.
[0267] In a yet further embodiment, casting takes place preferably
at a temperature between 50 and 60.degree. C., and the dosage is
performed through a nozzle connected to a valve attached to the
interior of the mixer. The pneumatically controlled valve opens for
the time necessary to let through the desired quantity of product,
the opening time of the valve is adjusted by a pneumatic timer and
the dosage is done in the same blister in which the product cools
down, determining the product shape.
[0268] The entire process can be done in a sterile environment,
using sterile materials, e.g., for the preparation of patches
without preservatives, since the ingredients used in the mixture
may provide a substrate for the proliferation of microorganisms
(similar polysaccharides are used in the culture broths to incubate
microorganisms in laboratory tests).
[0269] The process can also be carried out in an inert atmosphere
of nitrogen, in order to limit any possible oxidative processes on
the product.
[0270] The products obtained as described above can be used in both
cosmetics and therapeutic applications.
[0271] For cosmetic/therapeutic uses, the products of the invention
are appropriately formulated, possibly with other dermatologically
active substances.
[0272] Formulations appropriate to the purpose of this invention
include all types of cosmetic and dermopharmaceutical ingredients,
such as the molecules described herein.
[0273] In addition to the ingredients used to formulate the
products of this invention, the formulation may also contain other
biologically active ingredients and excipients, such as
surfactants, emollients, emulsifiers, solvents, moisturizers,
enhancers of percutaneous absorption, and in general all types of
ingredients, well-known to the formulators of cosmetic and
pharmaceutical products.
EXAMPLES
[0274] All examples in U.S. Pat. No. 7,458,982, U.S. Pat. No.
7,854,753, European patent EP1556061B2, and PCT publication WO
2009-124763 A2, including all associated data and figures
referenced therein, are hereby incorporate herein by reference.
[0275] Part of the Examples in WO 2009-124763 A2 are reproduced
below as Example 1.
Example 1
TABLE-US-00001 [0276] Ingredients INCI Name % Base formulation 1
Water Aqua up to 100 Methylpropanediol Methylpropanediol 10
Glycerin Glycerin 20 Chondrus crispus Chondrus crispus 2 Active
ingredient INCI name of active proper quantity ingredient
Preservative (if desired) Preservative systems proper quantity Base
formulation 2 Water Aqua up to 100 Butylene glycol Butylene glycol
10 Sorbitol solution 70% Sorbitol, Aqua 20 Chondrus crispus
Chondrus crispus 2 Active ingredient INCI name of active proper
quantity ingredient Preservative (if desired) Preservative systems
proper quantity
[0277] Examples of use of patches as carriers for active
ingredients are provided below.
[0278] Different patches have been prepared incorporating active
ingredients for cosmetic use. In an exemplary embodiment, the
method of preparation includes the following steps:
[0279] 1. Mix all components, adding the components in the order
shown above, heating at a temperature above 70.degree. C.,
preferably at 90.degree. C., under stirring.
[0280] 2. Cast the mixture in the blisters, keeping the temperature
of the nozzles preferably at around 50-60.degree. C.
[0281] 3. Close/seal the blister and allow to cool down to room
temperature.
[0282] More specifically, the production of the mask takes place in
a planetary mixer, equipped with a jacketing for heating with
temperature control (thermostat).
[0283] The preparation is conducted as follows: The product
Chondrus crispus is pre-dispersed in the mixture of glycerin and
methylpropanediol until it forms an homogeneous dispersion without
lumps. In the meantime, 75% of the water needed for the production
of the batch is heated to 90.degree. C.
[0284] After reaching this temperature, the suspension is added to
the water under vigorous stirring, then it is left to cool to
50-60.degree. C.
[0285] The water soluble active ingredient is dispersed in the
remainder of the water, and then added to the hot phase under
stirring, to obtain a complete dispersion. Finally the casting of
the product can be started.
[0286] The casting takes place preferably at the temperature of 50
and 60.degree. C., using a dosage system.
[0287] The dosage is performed by a nozzle connected to a valve,
attached to the interior of the mixer; the pneumatically controlled
valve opens for the time necessary to let through the desired
quantity of product. The opening time of the valve is adjusted by a
pneumatic timer.
[0288] The dosage is done in the same blister which, with the
cooling, imparts the shape to the product.
[0289] The product is then allowed to cool down, and an
aluminium/polythene tie layer sheet is applied to seal the
blister.
[0290] The blister is then die-cut, reaching the desired final
shape of the finished product.
[0291] A second exemplary patch contains sodium hyaluronate, as
described in table immediately below.
TABLE-US-00002 Components INCI Name % Water Aqua 67.5
Methylpropanediol Methylpropanediol 10 Glycerin Glycerin 20
Chondrus crispus Chondrus crispus 2 Sodium Hyaluronate (1800-2200
kDa) Sodium Hyaluronate 0.5 from Shandong Freda Biopharm
[0292] The method of preparation is similar to that of the previous
example immediately above. This patch has been shown to recover the
correct hydration level and is useful to treat the most
dry/dehydrated areas of the skin. The efficacy of this eye contour
patch has been tested successfully in clinical studies on
volunteers, where the hydrating and decongesting efficacy for this
area has been evaluated.
[0293] These gel patches have been used alone, and it has been
observed that there was a delivery of hydrosoluble active
ingredients to the skin in a time-controlled manner on the human
skin, by means of osmotic processes and occlusive effect.
Therefore, the invention products fulfill the requirements for
dermal masks, preferentially for face treatment.
[0294] There are several advantages associated with the present
invention respect to the common patches used for cosmetic and
dermopharmaceutical applications, for example: [0295] the matrix is
completely vegetal and biodegradable; [0296] the products show
optimal structural features: elasticity, flexibility, resistance,
adhesiveness, possibility to be removed and re-applied keeping the
original shape, transparency; [0297] there is no need for
pre-formed substrates like Tissue Non-Tissue (TNT) or polymers such
as polyester or polyacrylate, in which the desired active
ingredients are inserted and which require also an adhesive layer
for adhesion to the skin; [0298] the patch itself has hydrating
properties, thanks to its vegetable matrix structure; [0299] it is
possible to produce masks without preservatives and/or perfumes;
[0300] patches of any shape or size are easily obtained by casting
the melted mixture into a proper mould (blister); [0301] the mask
as such has an hydrating and lenitive effect to the skin; [0302]
the masks are dermatologically safe.
Example 2 IGF-1 In Vitro Testing
[0303] Insulin like growth factor I (IGF-1) is a 70 amino acid
polypeptide with 49% structural homology to insulin. The
intradermal deposition of IGF-1, (MW=7,676 Da) at a donor
concentration of 1.2 .mu.g/ml in propylene glycol. Intradermal drug
deposition for passive controls and photokinetic conditions was
determined at 15, 30, and 60 minutes.
[0304] Insulin like growth factor 1 (IGF-1) was analyzed using
commercially available ELISA kits according to manufacturer's
instructions (Kit # Diagnostic System Laboratories, Webster, Tex.).
Briefly, samples were incubated in 96-well plates coated with
specific primary antibody for periods of time, washed, incubated
with enzyme conjugate, washed again and then secondary HRP
conjugated antibodies were added. Color was developed using
tetramethylbenzidine substrate and the reaction was stopped by
adding sulfuric acid (0.2%). The plates were read on a microplate
reader (ELX800, BioTek Instruments, Winooski, Vt.) at 450 nm.
Standard curves were built for each analyte and the concentration
in each sample was calculated from standard curve.
[0305] FIG. 7 shows that IGF-1 was delivered into intact human skin
at significantly higher concentrations compared to passive
permeation, as measured at 15, 30 and 60 minutes. The data
demonstrates that the photokinetic system facilitates and increases
achievable tissue concentration of IGF-1 compared to passive
permeation.
Example 3 Human Growth Hormone In Vitro Testing
[0306] Human growth hormone (HGH) with a molecular weight of 22,124
Da at a final donor concentration of 270 ng/ml was prepared in a
formulation of 5% ethyl lactate, 30% propylene glycol, 1%
hyaluronic acid, 0.1% laurocapram, 0.1% Neolone.TM. 850, with the
balance as deionized water. Transdermal permeation under
photokinetic conditions of 350 nm, 390 nm, 405 nm, and 450 nm at 24
CPS were evaluated at 24 hours and compared with passive permeation
control samples.
[0307] Analytical assays performed by ELISA (Diagnostic System
Laboratories, Webster, Tex.) were analyzed using commercially
available kits according to manufacturer's instructions. Briefly,
samples were incubated in 96-well plates coated with specific
primary antibody for periods of time, washed, incubated with enzyme
conjugate, washed again and then secondary HRP conjugated
antibodies were added. Color was developed using
tetramethylbenzidine substrate, and the reaction was stopped by
adding sulfuric acid (0.2%). The plates were read on a microplate
reader (ELX800, BioTek Instruments, Winooski, Vt.) at 450 nm.
Standard curves were built for each analyte and the concentration
in each sample was calculated from standard curve.
[0308] FIG. 8 shows human growth hormone (HGH) transdermal
permeation quantities as measured at 24 hours exposed to 4
wavelengths of light pulsed at 24 cycles per second (24 cps). The
data demonstrates that the photokinetic system provides a
significantly increased achievable transdermal permeation fluxes of
HGH compared to passive permeation (p<0.001).
Example 4 Hyaluronic Acid In Vitro Testing
[0309] Hyaluronic acid (or hyaluronate) is an anionic, nonsulfated
glycosaminoglycan, and is available in various molecular weights.
Hyaluronic acid with a molecular weight 69,000 Daltons (69 kDa) at
a final donor concentration of 2% w/w, in a formulation of 5% ethyl
lactate, 30% propylene glycol, 1% hyaluronic acid, 0.1%
laurocapram, 0.1% Neolone.TM. 850, and with the balance as
deionized water, was assessed under various photokinetic conditions
for transdermal fluxes and intradermal deposition.
[0310] Analytical assays performed by ELISA (Diagnostic System
Laboratories, Webster, Tex.) were analyzed using commercially
available kits according to manufacturer's instructions. Briefly,
samples were incubated in 96-well plates coated with specific
primary antibody for periods of time, washed, incubated with enzyme
conjugate, washed again and then secondary HRP conjugated
antibodies were added. Color was developed using
tetramethylbenzidine substrate and the reaction was stopped by
adding sulfuric acid (0.2%). The plates were read on a microplate
reader (ELX800, BioTek Instruments, Winooski, Vt.) at 450 nm.
Standard curves were built for each analyte and the concentration
in each sample was calculated from standard curve.
[0311] FIG. 9A show transdermal permeation of 69 kDa hyaluronic
acid, determined under photokinetic conditions of 390 nm, 405 nm,
436 nm, and 450 nm light pulsed at 24 cycles per second (cps) and
100 cps compared to passive permeation at 24 hours. As shown for
this particular molecule, photokinetic transdermal flux rates using
405 nm light pulsed at 24 cps were significantly higher than
passive permeation (p<0.001). The flux rate achieved using 405
nm light pulsed at 24 cps was significantly higher than 405 nm
light pulsed at 100 cps (p<0.05).
[0312] FIG. 9B shows intradermal deposition of 69 kDa hyaluronic
acid, determined under photokinetic conditions of 390 nm, 405 nm,
436 nm, and 450 nm light pulsed at 24 cycles per second (cps) and
100 cps compared to passive deposition at 24 hours. Separate
controls were established for both the 24 cps and 100 cps groups.
As shown, for this particular molecule, the photokinetic condition
of 450 nm light pulsed at 100 cps produced significantly higher
intradermal deposition compared to the passive control
(p<0.05).
Example 5 Botulinum Toxin Type a In Vitro Testing
[0313] Botulinum toxin type A with a molecular weight of about 150
kDa (Botox.RTM. Allergan Irvine, Calif.) was dissolved in a
formulation comprising 5% ethyl lactate, 30% propylene glycol, 1%
hyaluronic acid, 0.1% Neolone.TM. 850 with the balance as deionized
water providing at a finale concentration of 2 units botulinum type
A per mL. The botulinum formulation was used in the photokinetic
testing apparatus previously described and evaluated at 405 nm and
450 nm for 24 and 100 cycles per second over 24 hours for both
transdermal and intradermal drug concentration.
[0314] Samples were analyzed with a fluorescent reporter assay
(Biosentinnel Pharmaceuticals, Madison Wis.) according to
manufacturer's instructions. Briefly, recipient chamber and tissue
extraction samples were incubated in 96-well plates with specific
primary antibody conjugated with a fluorescent enzyme for periods
of time, washed, incubated with substrate and the fluorescent
intensity was measured. Standard curves were built for each analyte
and the concentration in each sample was calculated from standard
curve. Transdermal fluxes are expressed as nanograms (ng)/cm.sup.2
of exposed skin/at 24 hours. Tissue deposition analysis
incorporated a measurement of the protein content in the extracted
samples as a method to normalize the extraction efficiency and are
expressed as units of botulinum toxin type A/mg protein/mg
skin.
[0315] Transdermal results for control passive permeation and 24
hour photokinetic exposure at 405 nm and 450 nm pulsed at 24 and
100 CPS are presented in FIG. 10A and FIG. 10B.
Example 6 Transdermal/Intradermal Delivery of 2,100 kDa Hyaluronic
Acid
[0316] Hyaluronic acid (or hyaluronate) is an anionic, nonsulfated
glycosaminoglycan and is available in various molecular weights.
Hyaluronic acid with a molecular weight of 2,100,000 Daltons (2,100
kDa) at a final donor concentration of 1% w/w in a formulation of
5% ethyl lactate, 30% propylene glycol, 1% hyaluronic acid, 0.1%
laurocapram, 0.1% Neolone.TM. 850, and with the balance as
deionized water, was assessed under various photokinetic conditions
for transdermal fluxes and intradermal deposition.
[0317] Analytical assays performed by ELISA were analyzed using
commercially available kits according to manufacturer's
instructions. Briefly, samples were incubated in 96-well plates
coated with specific primary antibody for periods of time, washed,
incubated with enzyme conjugate, washed again and then secondary
HRP conjugated antibodies were added. Color was developed using
tetramethylbenzidine substrate and the reaction was stopped by
adding sulfuric acid (0.2%). The plates were read on a microplate
reader (ELX800, BioTek Instruments, Winooski, Vt.) at 450 nm.
Standard curves were built for each analyte and the concentration
in each sample was calculated from standard curve.
[0318] Transdermal fluxes are expressed as nanograms (ng)/cm.sup.2
of exposed skin/at 24 hours.
[0319] Tissue deposition analysis incorporated a measurement of the
protein content in the extracted samples as a method to normalize
the extraction efficiency and are expressed as units of hyaluronic
acid/mg protein/mg skin.
[0320] FIG. 11A shows transdermal permeation of 2,100 kDa
hyaluronic acid determined under photokinetic conditions of 390 nm,
405 nm, 436 nm, and 450 nm light pulsed at 24 cycles per second
(cps) and 100 cps compared to passive permeation at 24 hours. As
shown for this particular molecule, photokinetic transdermal flux
rates using 390 nm, 405 nm, and 450 nm light pulsed at 24 cps, and
405 nm light pulsed at 100 cps were significantly higher than
passive permeation control samples (p<0.05). The flux rate
achieved using 390 nm, 405 nm, and 436 nm light pulsed at 24 cps
was significantly higher than 390 nm, 405 nm, and 436 nm light
pulsed at 100 cps (p<0.05).
[0321] FIG. 11B shows intradermal deposition of 2,100 kDa
hyaluronic acid determined under photokinetic conditions of 390 nm,
405 nm, 436 nm, and 450 nm light pulsed at 24 cycles per second
(cps) and 100 cps compared to passive deposition at 24 hours.
Separate controls were established for both the 24 cps and 100 cps
groups. As shown, for this particular molecule, the photokinetic
conditions of 436 nm and 450 nm light pulsed at 24 cps produced
significantly higher intradermal deposition compared to the passive
controls (p<0.05). 450 nm light pulsed at 24 cps demonstrated
significantly higher skin deposition than any of the light groups
pulsed at 100 cps (p<0.05).
Example 7 Intradermal Delivery of Hyaluronic Acid 2,180,000 Daltons
(2,180 kDa) at 15, 30 and 60 Minute Passive Control Vs.
Photokinetic Exposure of 3 Formulations
[0322] This example demonstrates the unexpectedly efficient
intradermal delivery of a biologically active substance (HA) using
a combination of the photokinetic method and the subject dermal
patch containing a red algae polysaccharide-based matrix (e.g., an
extract of Chondrus crispus at 2% by weight of the dermal
patch).
[0323] Gel Formulation
[0324] 1% hyaluronic acid, 63.9% citrus nobilis (fruit) extract,
35% propylene glycol with 0.1% preservative
[0325] Emulsion Cream Base
[0326] 1% hyaluronic acid in 87.9% Citrus nobilis (fruit) extract,
4% Olea Europaea Olive Fruit Oil, Hydrogenated Olive Oil, 2.5%
Sucrose Palmitate, 2% Squalene, 1% Cetearyl Alcohol, 0.5% Sucrose
Triesterate with 0.1% preservatives.
[0327] Patch Preparation as Per Example 1
[0328] 0.5% hyaluronic acid in a patch, as described above.
[0329] Analytical assays performed by ELISA were analyzed using
commercially available kits according to manufacturer's
instructions. Briefly, samples were incubated in 96-well plates
coated with specific primary antibody for periods of time, washed,
incubated with enzyme conjugate, washed again and then secondary
HRP conjugated antibodies were added. Color was developed using
tetramethylbenzidine substrate and the reaction was stopped by
adding sulfuric acid (0.2%). The plates were read on a microplate
reader (ELX800, BioTek Instruments, Winooski, Vt.) at 450 nm.
Standard curves were built for each analyte and the concentration
in each sample was calculated from standard curve.
[0330] Tissue deposition analysis incorporated a measurement of the
protein content in the extracted samples as a method to normalize
the extraction efficiency and are expressed as units of hyaluronic
acid/mg protein/mg skin at the 3 time points of 15, 30 and 60
minutes.
[0331] FIG. 12 shows intradermal deposition of 2,180 kDa hyaluronic
acid formulated in a gel (1% HA), a cream (1% HA), and a patch (at
about half the concentration of 0.5''/o), under photokinetic
conditions of 450 nm light pulsed at 100 cps at 15, 30, and 60
minutes of exposure compared to passive deposition controls taken
at the same time points. As shown, the passive deposition of the
gel and emulsion cream formulations produced neatly identical
deposition concentrations, while the patch formulation offered an
increased intradermal at the 15 and 60 minute time points. Under
photokinetic conditions of 450 nm light pulsed at 100 cps, all 3
formulations had increased depositions concentration compared to
passive controls at each time point. Surprisingly under
photokinetic conditions, the patch formulation produced higher
intradermal deposition compared to the gel or emulsion cream
formulations.
[0332] Hyaluronic acid normally occurs within skin. Samples of skin
not exposed to the topical hyaluronic acid formulations
demonstrated a level of 3,385 nanograms/milligram of skin. The
three formulations produced increased skin deposition compared to
normally occurring hyaluronic acid within the skin.
[0333] It is apparent, based on FIG. 12, that patch-mediated
delivery of HA (at about half the concentration of 0.5%) is
surprisingly more effective (about 200-300% more effective)
compared to cream- or gel-mediated HA delivery, even though similar
photokinetic parameters were used in the experiments.
Example 8 Hyaluronic Acid Patch on a 55 Year Old Male Forehead
Wrinkle
[0334] One half of a prominent forehead wrinkle was covered with
the patch and was exposed to 450 nm light at 100 CPS for one hour.
After one hour the patch was removed and the two halves of the same
wrinkle were evaluated. It was immediately obvious that the exposed
forehead area had a pronounced reduction in wrinkle depth and
prominence.
REFERENCES
[0335] Barrak Al-Qallaf, Diganta Bhusan Das, Daisuke Mori and
Zhanfeng Cui, Modeling transdermal delivery of high molecular
weight drugs from microneedle systems, Phil. Trans. R. Soc. A 365,
2951-2967, 2007. [0336] Beatrice M. Magnusson, Yuri G. Anissimov,
Sheree E. Cross, and
[0337] Michael S. Roberts, Molecular Size as the Main Determinant
of Solute Maximum Flux Across the Skin, J Invest Dermatol 122:
993-999, 2004. [0338] Eseldin Kelebl, Rakesh Kumar Sharma, Esmaeil
B mosa, Abd-alkadar Z aljahwi, Transdermal Drug Delivery
System--Design and Evaluation International Journal of Advances in
Pharmaceutical Sciences 1: 201-211, 2010. [0339] Ritesh Kumar and
Anil Philip, Modified Transdermal Technologies: Breaking the
Barriers of Drug Permeation via the Skin, Tropical Journal of
Pharmaceutical Research, 6(1): 633-644, 2007.
[0340] All references cited are incorporated herein by
reference.
TABLE-US-00003 Annex VI (Part 1) List of preservatives allowed
Conditions of use and warnings which Reference Maximum authorized
must be number Substance concentration Limitations and requirements
printed on the label a b c d e 1a Salts of benzoic acid other than
0.5% (acid) that listed under reference number 1 and esters of
benzoic acid 1 Benzoic acid (CAS No 65-85-0) Rinse-off products,
except oral and its sodium salt (CAS No care products: 532-32-1)
2.5% (acid) Oral care products: 1.7% (acid) Leave-on products: 0.5%
(acid) 2 Propionic acid and its salts 2% (acid) 3 Salicylic acid
and its salts (*) 0.5% (acid) Not to be used in preparations Not to
be used for for children under three years of children under three
age, except for shampoos years of age (1) 4 Sorbic acid
(hexa-2,4-dienoic 0.6% (acid) acid) and its salts 5 Formaldehyde
and 0.2% (except for oral hygiene) Prohibited in aerosol
dispensers, paraformaldehyde (*) 0.1% (for oral hygiene) except for
foams concentrations expressed as free formaldehyde 7 Biphenyl-2-ol
(o-phenylphenol) 0.2% expressed as the phenol and its salts 8 Zinc
pyrithione (*) (CAS No Hair products: 1.0% Rinse-off products only
No use 13463-41-7) Other products: 0.5% in products for oral
hygiene 9 Inorganic sulphites and 0.2% expressed as free SO2
hydrogensulphites (*) 11 Chlorobutanol (INN) 0.5% Prohibited in
aerosol dispensers Contains chlorobutanol (sprays) 12
4-hydroxybenzoic acid and its 0.4% (acid) for 1 ester salts and
esters 0.8% (acid) for mixtures of esters 13
3-Acetyl-6-methylpyran- 0.6% (acid) Prohibited in aerosol
dispensers 2,4(3H)-dione (Dehydroacetic (sprays) acid) and its
salts 14 Formic acid and its sodium salt 0.5% (expressed as acid)
15 3,3'-Dibromo-4,4'- 0.1% hexamethylenedioxydibenzamidine
(Dibromohexamidine) and its salts (including isethionate) 16
Thiomersal (INN) 0.007% (of Hg). For eye make-up and eye make-
Contains thiomersal If mixed with other mercurial up remover only
authorized by this Directive, the maximum concentration of Hg
remains fixed at 0.007% 17 Phenylmercuric salts (including 0.007%
(of Hg). For eye make-up and eye make- Contains borate) If mixed
with other mercurial up remover only phenylmercuric compounds
authorized by this compounds Directive, the maximum concentration
of Hg remains fixed at 0.007% 18 Undec-10-enoic acid and its 0.2%
(acid) salts 19 Hexetidine (INN) 0.1% Rinse-off products only 20
5-Bromo-5-nitro-1,3-dioxane 0.1% Rinse-off products only Avoid
formation of nitrosamines 21 Bronopol (INN) 0.1% Avoid formation of
nitrosamines 22 2,4-Dichlorobenzyl alcohol 0.15% 23 Triclocarban
(INN) (*) 0.2% Purity criteria: 3,3',4,4'- Tetrachloroazobenzene
<1 ppm; 3,3',4,4'- Tetrachloroazoxybenzene <1 ppm 24
4-Chloro-m-cresol 0.2% Prohibited in the products intended to come
into contact with mucous membranes 25 Triclosan (INN) 0.3% 26
4-Chloro-3,5-xylenol 0.5% 27 3,3'-Bis(1-hydroxymethyl-2,5- 0.6%
dioxoimidazolidin-4-yl)-1,1'- methylenediurea ("Imidazolidinyl
urea") 28 Poly(1-hexamethylenebiguanide 0.3% hydrochloride) 29
2-Phenoxyethanol 1.0% 30 Hexamethylenetetramine 0.15% (methenamine)
(INN) 31 Methenamine 3- 0.2% chloroallylochloride (INNM) 32
1-(4-Chlorophenoxy)-1- 0.5% (imidazol-1-yl)-3,3-dimethylbutan-
2-one 33 1,3-Bis(hydroxymethyl)-5,5- 0.6%
dimethylimidazolidine-2,4-dione 34 Benzyl alcohol (*) 1.0% 35
1-Hydroxy-4-methyl-6-(2,4,4- 1% rinse-off products
trimethylpentyl)-2 pyridon and its 0.5% other products
monoethanolamine salt 37 6,6-Dibromo-4,4-dichloro-2,2'- 0.1%
methylene-diphenol (Bromochlorophen) 38 4-Isopropyl-m-cresol 0.1%
39 Mixture of 0.0015% (of a mixture in the
5-Chloro-2-methyl-isothiazol-3(2H)-one ratio 3:1 of and
5-Chloro-2-methyl- 2-Methylisothiazol-3(2H)-one with
isothiazol-3(2H)-one and magnesium 2-Methylisothiazol-3(2H)-
chloride and magnesium nitrate one 40 2-Benzyl-4-chlorophenol
(chlorophene) 0.2% 41 2-Chloroacetamide 0.3% Contains 42
Chlorhexidine (INN) and its 0.3% expressed as chloroacetamide
digluconate, diacetate and chlorhexidine dihydrochloride 43
1-Phenoxypropan-2-ol (*) 1.0% Only for rinse-off products 44 Alkyl
(C12-C22) trimethyl ammonium, 0.1% bromide and chloride (*) 45
4,4-Dimethyl-1,3-oxazolidine 0.1% The pH of the finished product
must not be lower than 6 46 N-(Hydroxymethyl)-N- 0.5%
(dihydroxymethyl-1,3-dioxo- 2,5-imidazolidinyl-4)-N'-
(hydroxymethyl)urea 47 1,6-Di(4-amidinophenoxy)-n- 0.1% hexane
(Hexamidine) and its salts (including isothionate and p-
hydroxybenzoate) 48 Glutaraldehyde (Pentane-1,5-dial) 0.1%
Prohibited in aerosols (sprays) Contains glutaraldehyde (where
glutaraldehyde concentration in the finished product exceeds 0.05%)
49 5-Ethyl-3,7-dioxa-1- 0.3% Prohibited in oral hygiene
azabicyclo[3.3.0] octane products and in products intended to come
into contact with mucous membranes 50
3-(p-Chlorophenoxy)-propane-1,2- 0.3% diol (chlorphenesin) 51
Sodium hydroxymethylamino acetate 0.5% (Sodium
hydroxymethylglycinate) 52 Silver chloride deposited on 0.004%
calculated as AgCl 20% AgCl (w/w) on TiO2 titanium dioxide
Prohibited in products for children under three years of age, in
oral hygiene products and in products intended for application
around the eyes and on the lips 53 Benzethonium Chloride (INCI)
0.1% (a) Rinse-off products (b) Leave-on products other than for
oral care use 54 Benzalkonium chloride, bromide 0.1% calculated as
Avoid contact and saccharinate (*) benzalkonium chloride with eyes
55 Benzylhemiformal 0.15% Only for products to be removed by
rinsing 56 Iodopropynyl butylcarbamate; (a) Rinse-off products:
0.02% Not to be used in oral hygiene and (a) Not to be used for
(IPBC); (b) Leave-on products: lip care products. children under
three 3-Iodo-2-propynylbutylcarbamate 0.01%, except in (a) Not to
be used in preparations years of age (**) CAS No: 55406-53-6
deodorants/antiperspirants: for children under three years of (b)
Not to be used for 0.0075% age except in bath products/shower
children under three gels and shampoos years of age (***) (b) Not
to be used in body lotion and body cream (*) Not to be used in
preparations for children under three years of age 57
Methylisothiazolinone (INCI) 0.01% 58 Ethyl Lauroyl Arginate HCl
0.4% Not to be used in lip products, oral (INCI) (*) (**) products
and spray products Ethyl-N-alpha-dodecanoyl-L- arginate
hydrochloride CAS No 60372-77-2 EC No 434-630-6 PREAMBLE 1.
Preservatives are substances which may be added to cosmetic
products for the primary purpose of inhibiting the development of
micro-organisms in such products. 2. The substances marked with the
symbol (*) may also be added to cosmetic products in concentrations
other than those laid down in this Annex for other specific
purposes apparent from the presentation of the product, e.g. as
deodorants in soaps or as anti-dandruff agents in shampoo. 3. Other
substances used in the formulation of cosmetic products may also
have anti-microbial properties and thus help in the preservation of
the products, as, for instance, may essential oils and some
alcohols. These substances are not included in this Annex. 4. For
the purposes of this list: "Salts" is taken to mean: salts of the
cations sodium, potassium, calcium, magnesium, ammonium and
ethanolamines: salts of the anions chloride, bromide, sulphate,
acetate. "Esters" is taken to mean: esters of methyl, ethyl,
propyl, isopropyl, butyl, isobutyl, phenyl. 5. All finished
products containing formaldehyde or substances in this Annex and
which release formaldehyde must be labelled with the warning
"contains formaldehyde" where the concentration of formaldehyde in
the finished product exceeds 0.05%. NOTES (1) Solely for products
which might be used for children under three years of age and which
remain in prolonged contact with the skin (*) Concerns any products
aimed to be applied on a large part of the body (**) Solely for
products, other than bath products/shower gels and shampoo, which
might be used for children under three years of age (***) Solely
for products which might be used for children under three years of
age
* * * * *
References