U.S. patent application number 16/898365 was filed with the patent office on 2021-04-29 for absorption enhancers for drug administration.
The applicant listed for this patent is AEGIS THERAPEUTICS, LLC. Invention is credited to Edward T. Maggio.
Application Number | 20210121573 16/898365 |
Document ID | / |
Family ID | 1000005326577 |
Filed Date | 2021-04-29 |
United States Patent
Application |
20210121573 |
Kind Code |
A1 |
Maggio; Edward T. |
April 29, 2021 |
ABSORPTION ENHANCERS FOR DRUG ADMINISTRATION
Abstract
The present invention provides compositions and methods and for
increasing the bioavailability of therapeutic agents in a subject.
The compositions include at least one alkyl glycoside and at least
one therapeutic agent, wherein the alkylglycoside has an alkyl
chain length from about 10 to about 16 carbon atoms. In various
aspects, the invention provides compositions and methods for oral
delivery in the form of a tablet.
Inventors: |
Maggio; Edward T.; (San
Diego, CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
AEGIS THERAPEUTICS, LLC |
San Diego |
CA |
US |
|
|
Family ID: |
1000005326577 |
Appl. No.: |
16/898365 |
Filed: |
June 10, 2020 |
Related U.S. Patent Documents
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Application
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Filing Date |
Patent Number |
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16352525 |
Mar 13, 2019 |
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16898365 |
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15942110 |
Mar 30, 2018 |
10265402 |
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16352525 |
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15655669 |
Jul 20, 2017 |
10512694 |
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15942110 |
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13951284 |
Jul 25, 2013 |
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15655669 |
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13191146 |
Jul 26, 2011 |
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13951284 |
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12906922 |
Oct 18, 2010 |
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13191146 |
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12341696 |
Dec 22, 2008 |
8268791 |
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12906922 |
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12195192 |
Aug 20, 2008 |
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12341696 |
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12036963 |
Feb 25, 2008 |
8642564 |
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12195192 |
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11193825 |
Jul 29, 2005 |
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12036963 |
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11127786 |
May 11, 2005 |
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11193825 |
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60649958 |
Feb 3, 2005 |
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60637284 |
Dec 17, 2004 |
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60632038 |
Nov 30, 2004 |
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60609890 |
Sep 14, 2004 |
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60604296 |
Aug 25, 2004 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 47/26 20130101;
A61K 31/4439 20130101; A61K 9/0043 20130101; A61K 9/006 20130101;
A61K 9/2095 20130101; A61K 9/0048 20130101; A61K 9/0056 20130101;
A61K 31/138 20130101; A61K 9/2018 20130101; A61K 31/70 20130101;
A61K 31/137 20130101 |
International
Class: |
A61K 47/26 20060101
A61K047/26; A61K 31/137 20060101 A61K031/137; A61K 31/138 20060101
A61K031/138; A61K 9/00 20060101 A61K009/00; A61K 9/20 20060101
A61K009/20; A61K 31/4439 20060101 A61K031/4439; A61K 31/70 20060101
A61K031/70 |
Claims
1-7. (canceled)
8. A method of modulating insulin levels in a subject, said method
comprising: administering, to said subject in need of insulin
modulation, a pharmaceutical composition comprising: glucagon-like
peptide-1 (GLP-1), a peptide thereof, or any analog thereof, and
about 0.2% w/v to about 1% w/v of alkyl glycoside.
9. The method of claim 8, wherein the subject has diabetes.
10. The method of claim 9, wherein the diabetes is selected from
brittle diabetes, chemical diabetes, impaired glucose tolerance,
gestational diabetes, diabetes insipidus, diabetes insipidus
central, diabetes insipidus nephrogenic, diabetes insipidus
pituitary, latent diabetes, lipatrophic diabetes, maturity-onset
diabetes of youth (MODY), diabetes mellitus (DM), diabetes mellitus
adult-onset (type 2 DM), diabetes mellitus insulin-dependent (type
1 DM), diabetes mellitus non-insulin dependent (NIDDM), diabetes
mellitus juvenile or juvenile-onset, diabetes mellitus
ketosis-prone, diabetes mellitus ketosis-resistant, diabetes
mellitus malnutrition-related (MRDM), diabetes mellitus tropical or
tropical pancreatic, diabetes mellitus, preclinical diabetes, or
drug induced diabetes.
11. The method of claim 10, wherein the diabetes is type 1 DM.
12. The method of claim 10, wherein the diabetes is type 2 DM.
13. The method of claim 8, wherein the alkyl glycoside is selected
from the group consisting of dodecyl maltoside, tridecyl maltoside,
tetradecyl maltoside, sucrose mono-dodecanoate, sucrose
mono-tridecanoate, and sucrose mono-tetradecanoate.
14. The method of claim 13, wherein the alkyl glycoside is dodecyl
maltoside.
15. The method of claim 8, wherein the pharmaceutical composition
comprises about 0.2% w/v to about 0.5% w/v of the alkyl
glycoside.
16. The method of claim 8, wherein the alkyl glycoside is dodecyl
maltoside and the pharmaceutical composition comprises 0.2% w/v to
about 0.5% w/v of dodecyl maltoside.
17. The method of claim 8, wherein the pharmaceutical composition
comprises GLP-1.
18. The method of claim 8, wherein the pharmaceutical composition
comprises a GLP-1 analog selected from the group consisting of
diaglutide, albiglutide, taspoglutide, lixinsenatide, exenatide,
and liraglutide.
19. The method of claim 17, wherein the GLP-1 analog is
liraglutide.
20. The method of claim 8, wherein the pharmaceutical composition
is formulated for intranasal administration and said intranasal
formulation is administered to a nasal mucosal membrane of the
subject.
21. The method of claim 20, wherein about 50 .mu.L to about 200
.mu.L of the intranasal formulation is administered to the nasal
mucosal membrane of the subject.
22. The method of claim 20, wherein the intranasal formulation is
in a format selected from a drop, a spray, an or an aerosol.
23. The method of claim 8, further comprising administering an
agent that inhibits protein degradation to the subject.
24. The method of claim 23, wherein the agent that inhibits protein
degradation is a protease or peptidase inhibitor.
25. The method of claim 8, wherein the composition further
comprises a stabilizing agent.
26. The method of claim 25, wherein the GLP-1, peptide thereof, or
analog thereof is bound to said stabilizing agent.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a continuation application of U.S.
application Ser. No. 16/352,525 filed Mar. 13, 2019, now pending;
which is a continuation application of U.S. application Ser. No.
15/942,110 filed Mar. 30, 2018, now issued as U.S. Pat. No.
10,265,402; which is a continuation application of U.S. application
Ser. No. 15/655,669 filed Jul. 20, 2017, now issued as U.S. Pat.
No. 10,512,694; which is a continuation application of U.S.
application Ser. No. 13/951,284 filed Jul. 25, 2013, now abandoned;
which is a continuation-in-part of U.S. application Ser. No.
13/191,146 filed Jul. 26, 2011, now abandoned; which is a
continuation-in-part of U.S. application Ser. No. 12/906,922 filed
Oct. 18, 2010, now abandoned; which is a continuation-in-part of
U.S. application Ser. No. 12/341,696 filed Dec. 22, 2008, now
issued as U.S. Pat. No. 8,268,791; which is a continuation-in-part
of U.S. application Ser. No. 12/195,192 filed Aug. 20, 2008, now
abandoned, which is a continuation-in-part of U.S. application Ser.
No. 12/036,963 filed Feb. 25, 2008, now issued as U.S. Pat. No.
8,642,564; which is a continuation-in-part of U.S. application Ser.
No. 11/193,825 filed Jul. 29, 2005, now abandoned, which is a
continuation-in-part of U.S. application Ser. No. 11/127,786 filed
May 11, 2005, now abandoned; which claims the benefit under 35 USC
.sctn. 119(e) to U.S. Application Ser. No. 60/649,958 filed Feb. 3,
2005, U.S. Application Ser. No. 60/637,284 filed Dec. 17, 2004,
U.S. Application Ser. No. 60/632,038 filed Nov. 30, 2004, U.S.
Application Ser. No. 60/609,890 filed Sep. 14, 2004 and to U.S.
Application Ser. No. 60/604,296 filed Aug. 25, 2004, all now
expired. The disclosure of each of the prior applications is
considered part of and is incorporated by reference in the
disclosure of this application.
BACKGROUND OF THE INVENTION
Field of the Invention
[0002] The present invention relates to generally to compositions
and solid oral dosage forms containing a pharmaceutically active
ingredient and an alkylsaccharide, which enhances the oral
bioavailability for the absorption of the active ingredient.
Background Information
[0003] Therapeutic agents are often combined with various
surfactants. Yet, surfactants are frequently irritating to the skin
and other tissues, including mucosal membranes such as those found
in the nose, mouth, eye, vagina, rectum, esophagus, intestinal
tract, and the like. Many surfactants also cause proteins to
denature, thus destroying their biological activity. Another
serious limitation to the development and use of such agents is the
ability to deliver them safely, non-invasively, efficiently and
stably to the site of action. Therefore, an ideal enhancing
surfactant will stabilize the therapeutic agent, be non-toxic and
non-irritable to the skin or mucosal surfaces, have antibacterial
activity, and enhance the passage or absorption of the therapeutic
agent through various membrane barriers without damaging the
structural integrity and biological function of the membrane and
increase bioavailability of the agent.
[0004] In spite of the many attractive aspects of peptides and
proteins as potential therapeutic agents, their susceptibility to
denaturation, hydrolysis, and poor absorption in the
gastrointestinal tract makes them unsuitable for oral
administration, typically requiring administration by injection.
This remains a major shortcoming. Compared to small molecule drugs,
peptides are considerably less stable. Careful attention must be
paid to formulation and storage to avoid unwanted degradation. Some
proteins, particularly proteins with substantially non-naturally
occurring amino acid sequences can be immunogenic. Upon injection,
immune cells may be recruited to the site of injection and a
humoral or cellular immune response may be induced. Aggregated
peptides are known to be more prone to eliciting an immunogenic
response than monomers. This may be avoided to a greater to or
lesser extent if the peptide can be directly absorbed from the
gastrointestinal tract into systemic circulation. Therefore, while
the range of clinical indications for therapeutic proteins and
peptides is quite broad, the actual number of such therapeutics in
general use today is quite small compared to the number of
chemically synthesized and orally active pharmaceuticals currently
on the market. In recent years, development of a large class of
alkylsaccharide delivery enhancement agents, for example, molecules
that provide intranasal bioavailabilities, comparable to those
achieved by injection have been investigated. While recent
developments in intranasal delivery for proteins and peptides are
creating new and expanded opportunities for practical clinical uses
of peptides, proteins, and other macromolecular therapeutics, few,
if any, peptides appear to be administrable orally due to
unacceptably low oral bioavailability. A number of studies have
been conducted to demonstrate oral bioavailability for a variety of
peptide drugs. These studies used a variety of absorption enhancers
as well as physical processes such as micronization. For example
among formulations specifically optimized for oral delivery,
insulin exhibited only 3% oral bioavailability (Badwin et al.,
2009). Calcitonin exhibited only 0.5-1.4% oral bioavailability
(Bucklin 2002). Parathyroid hormone has been shown to exhibit 2.1%
oral bioavailability (Leone-Bay et al., 2001). There are two
principal biochemical problems limiting the oral absorption of
peptides. The first relates to the susceptibility of peptides to
hydrolysis in the gastrointestinal tract. The second relates to
intrinsically poor absorption across the intestinal mucosal
membrane.
[0005] Incorporation of non-standard amino acids into peptide
sequences has been shown to reduce hydrolysis or slow metabolism
for some peptides. Non-standard aminoacyl residues have been
incorporated into a number of drugs for this purpose allowing the
drugs to remain active for a longer period of time than otherwise
possible. Non-standard amino acids are those amino acids that are
not among the 22 naturally occurring L-amino acids found in
proteins. There exist a vast number of non-standard amino acids
that may be considered for such use in either the D or L
configuration. A few examples include, but are not limited to,
allylglycine, (2S,3R,4S)-.alpha.-(carboxycyclopropyl)glycine,
.alpha.-cyclohexylglycine, C-propargylglycine,
.alpha.-neopentylglycine, .alpha.-cyclopropylglycine,
N-lauroylsarcosine sodium salt, N-(4-hydroxyphenyl)glycine,
N-(2-furoyl)glycine, naphthylglycine, phenylglycine, lanthionine,
2-aminoisobutyric acid, dehydroalanine, gamma-aminobutyric
acid.
[0006] Some specific examples of non standard amino acids used in
drugs include D-4-hydroxyphenylglycine which is incorporated into
the antibacterial drug Amoxicillin, D-phenylglycine which is
incorporated into the antihypertensive drug Enalapril, and
(2R,3S)-phenylisoserine which is incorporated into the
antineoplastic drug Taxol.
[0007] In the case of peptide drugs, D-2-Naphthylalanine is
incorporated into the endometriosis drug Nafarelin. The D-isomers
of naturally occurring L-amino acids are frequently used to
increase stability of peptide drugs. Examples of D-amino acid
stabilized peptides include the anti-obesity peptide D-Leu-OB3 (Lee
et al., 2010) and the CCR5 anti-HIV drug D-ala-peptide T (DAPTA)
(Ruff et al., 2001) among others.
[0008] Enzymatic hydrolysis in the gastrointestinal tract may also
be reduced or eliminated by addition of specific enzyme inhibitors
such as bacitracin, bestatin, amastatin, boroleucin, borovaline,
aprotinin, and trypsin inhibitor among others.
[0009] Alkylsaccharides have been demonstrated to enhance oral
absorption of small molecules, and peptides, when presented as
aqueous solutions of the alkylsaccharide and the small molecule or
peptide. In such solutions, the concentration of the
alkylsaccharide is higher than the critical micelle concentration
(CMC). An example includes the oral delivery of octreotide in
aqueous solution using dodecyl-beta-D-maltoside as the
alkylsaccharide absorption enhancer. Another example is oral
delivery of a leptin-related synthetic peptide insulin sensitizer
using an aqueous dodecyl maltoside solution. Yet another example is
oral delivery of exenatide and pramlintide using
dodecyl-beta-D-maltoside in an aqueous solution as the
alkylsaccharide absorption enhancer. Yet another example is oral
delivery of heparin using tetradecyl maltoside in an aqueous
solution as the alkyl saccharide absorption enhancer (see for
example, Maggio and Grasso, Regulatory Peptides 167 (2011) 233-238;
Novakovic, et al., Peptides, 43 (2013) 167-163; Leinung M C et al.,
Regulatory Peptides 179:33-38 (2012); Yang, et al., (2005) Journal
of Drug Targeting, 13:1, 29-38).
SUMMARY OF THE INVENTION
[0010] The present invention is based, in part, on the development
of a therapeutic composition containing a drug enhancing agent
useful for increasing the absorption and bioavailability of the
drug, while at the same time avoiding various adverse toxic effects
of drug. In particular, the drug enhancing agents of the invention
contain a non-toxic surfactant consisting of at least an alkyl
glycoside and/or saccharide alkyl ester. One advantage of the
therapeutic compositions of the invention is that they permit
administration and delivery of the therapeutic agents with high
bioavailabilities at concentrations of enhancing agents that are
dramatically below their so-called "no observable adverse effect
levels" (their NOAEL's). Accordingly, the present invention
provides compositions, including alkyl glycosides and/or saccharide
alkyl esters and a therapeutic agent (e.g., small molecule organic
drug molecules, low molecular weight peptides such as Exenatide,
GLP-1 and the like, proteins, and non-peptide therapeutic polymers
such as low molecular weight heparin and inhibitory RNA), methods
of administering and using the compositions e.g., via the oral,
ocular, nasal, nasolacrimal, inhalation or pulmonary, oral cavity
(sublingual or Buccal cell) or cerebral spinal fluid (CSF) delivery
route, and methods of ameliorating a disease state in a subject by
administration of such compositions.
[0011] Previously, no examples of the use of alkylsaccharides as
absorption enhancers in solid dosage forms such as tablets has been
demonstrated. While alkylsaccharides such as dodecyl maltoside,
n-tetradecyl maltoside have relatively high levels of solubility in
aqueous solution, they dissolve only very slowly and because a
significant portion of the molecules are comprised of the
hydrophobic linear alkyl chains. In order to ensure complete
dissolution of these alkylsaccharides in aqueous solution, the
aqueous solution is mildly heated and the container holding the
aqueous media and the alkylsaccharide is either stirred or agitated
continuously for up to 15 to 30 min. in order to ensure complete
dissolution of the alkylsaccharide. As a result, it was not
expected that solid dosage forms containing an alkylsaccharide and
the drug substance would benefit from the absorption enhancing
effect of the alkylsaccharide since the slow dissolution of the
alkylsaccharide in the aqueous gastrointestinal contents would not
be expected to produce a sufficiently high concentration or
comparable concentration to that achieved in the aqueous
formulations before the drug substance and the alkylsaccharide
molecules diffuse away from each other throughout the
gastrointestinal contents diluting the relative concentrations of
each. Surprisingly, solid dosage forms comprising an
alkylsaccharide, and a pharmacologically active substance, along
with other inactive excipients formed into a tablet were found to
provide a substantial increase in oral bioavailability of the
pharmacologically active substance, as shown in the examples. In
addition to active pharmaceutical substance and alkylsaccharide
absorption enhancer, other inactive excipients may include by way
of example candelilla wax, hypromellose, magnesium stearate,
microcrystalline cellulose, polyethylene glycol, povidone, and
titanium dioxide.
[0012] In one aspect, the present invention relates to a surfactant
composition having at least one alkyl glycoside and/or at least one
saccharide alkyl ester, and when admixed, mixed or blended with a
therapeutic agent, a drug, or biologically active compound, the
surfactant stabilizes the biological activity and increases the
bioavailability of the drug.
[0013] Accordingly, in one aspect, the invention provides a
therapeutic composition having at least one biologically active
compound and at least one surfactant, wherein the surfactant
further consists of at least one alkyl glycoside and/or saccharide
alkyl ester or sucrose ester and wherein the therapeutic
composition stabilizes the biologically active compound for at
least about 6 months, or more, and from about 4.degree. C. to about
25.degree. C.
[0014] The invention also provides a method of administering a
therapeutic composition having a surfactant including at least one
alkyl glycoside and/or saccharide alkyl ester admixed, mixed, or
blended with at least one therapeutic agent, or a drug, or
biologically active compound, and administered or delivered to a
subject, wherein the alkyl has from about 10 to 24, 10 to 20, 10 to
16, or 10 to 14 carbon atoms, wherein the surfactant increases the
stability and bioavailability of the therapeutic agent.
[0015] In yet another aspect, the invention provides a method of
increasing absorption of a low molecular weight compound into the
circulatory system of a subject by administering the compound via
the oral, ocular, nasal, nasolacrimal, inhalation or pulmonary,
oral cavity (sublingual or Buccal cell), or CSF delivery route when
admixed, mixed or blended with an absorption increasing amount of a
suitable surfactant, wherein the surfactant is a nontoxic and
nonionic hydrophobic alkyl joined by a linkage to a hydrophilic
saccharide. Such low molecular weight compounds include but are not
limited to, nicotine, interferon, PYY, GLP-1, synthetic exendin-4,
parathyroid hormone, human growth hormone, or a small organic
molecule. Additional low molecular weight compounds include
antisense oligonucleotides or interfering RNA molecules (e.g.,
siRNA or RNAi).
[0016] The present invention also provides a method of treating
diabetes including administering to a subject in need thereof via
the oral, ocular, nasal, nasolacrimal, inhalation or pulmonary, or
oral cavity (sublingual or Buccal cell), a blood glucose reducing
amount of a therapeutic composition, for example, an incretin
mimetic agent or a functional equivalent thereof, and an absorption
increasing amount of a suitable nontoxic, nonionic alkyl glycoside
having a hydrophobic alkyl group joined by a linkage to a
hydrophilic saccharide, thereby increasing the absorption of
incretin mimetic agent or insulin and lowering the level of blood
glucose and treating diabetes in the subject.
[0017] The present invention also provides a method of treating
congestive heart failure in a subject including administering to
the subject in need thereof via the oral, ocular, nasal,
nasolacrimal, or inhalation delivery route, a therapeutically
effective amount of a composition comprising a GLP-1 peptide or a
functional equivalent thereof, and an absorption increasing amount
of a suitable nontoxic, nonionic alkyl glycoside having a
hydrophobic alkyl joined by a linkage to a hydrophilic saccharide,
thereby treating the subject.
[0018] In another aspect, the invention provides a method of
treating obesity or diabetes associated with obesity in a subject
comprising administering to a subject in need thereof via the oral,
ocular, nasal, nasolacrimal, inhalation or CSF delivery route, a
therapeutically effective amount of a composition comprising a PYY
peptide or a functional equivalent thereof, and an absorption
increasing amount of a suitable nontoxic, nonionic alkyl glycoside
having a hydrophobic alkyl joined by a linkage to a hydrophilic
saccharide, thereby treating the subject.
[0019] In another aspect, the invention provides a method of
increasing absorption of a low molecular weight therapeutic
compound into the circulatory system of a subject by administering
via the oral, ocular, nasal, nasolacrimal, inhalation or CSF
delivery route the compound and an absorption increasing amount of
a suitable nontoxic, nonionic alkyl glycoside having a hydrophobic
alkyl group joined by a linkage to a hydrophilic saccharide,
wherein the compound is from about 1-30 kD, with the proviso that
the compound is not insulin, calcitonin, or glucagon when the route
of administration is oral, ocular, nasal, or nasolacrimal.
[0020] The present invention also provides a method of increasing
absorption of a low molecular weight therapeutic compound into the
circulatory system of a subject by administering via the oral,
ocular, nasal, nasolacrimal, inhalation or pulmonary, oral cavity
(sublingual or Buccal cell) or CSF delivery route the compound and
an absorption increasing amount of a suitable nontoxic, nonionic
alkyl glycoside having a hydrophobic alkyl joined by a linkage to a
hydrophilic saccharide, wherein the compound is from about 1-30
kilo Daltons (kD), with the proviso that the subject does not have
diabetes when delivery is via the oral, ocular, nasal or
nasolacrimal routes.
[0021] In one aspect of the invention, there is provided a
pharmaceutical composition having a suitable nontoxic, nonionic
alkyl glycoside having a hydrophobic alkyl group joined by a
linkage to a hydrophilic saccharide in combination with a
therapeutically effective amount of Exenatide (exendin-4) in a
pharmaceutically acceptable carrier.
[0022] In one aspect, the invention provides a pharmaceutical
composition having a suitable nontoxic, nonionic alkyl glycoside
having a hydrophobic alkyl group joined by a linkage to a
hydrophilic saccharide in combination with a therapeutically
effective amount of GLP-1 in a pharmaceutically acceptable
carrier.
[0023] In one aspect, the invention provides a pharmaceutical
composition having a suitable nontoxic, nonionic alkyl glycoside
having a hydrophobic alkyl group joined by a linkage to a
hydrophilic saccharide in combination with a therapeutically
effective amount of nicotine in a pharmaceutically acceptable
carrier.
[0024] In one aspect, the invention provides a pharmaceutical
composition comprising a suitable nontoxic, nonionic alkyl
glycoside having a hydrophobic alkyl group joined by a linkage to a
hydrophilic saccharide in combination with a therapeutically
effective amount of interferon in a pharmaceutically acceptable
carrier.
[0025] In one aspect, the invention provides pharmaceutical
composition having a suitable nontoxic, nonionic alkyl glycoside
having a hydrophobic alkyl group joined by a linkage to a
hydrophilic saccharide in combination with a therapeutically
effective amount of PYY in a pharmaceutically acceptable
carrier.
[0026] In one aspect, the invention provides a pharmaceutical
composition having a suitable nontoxic, nonionic alkyl glycoside
having a hydrophobic alkyl group joined by a linkage to a
hydrophilic saccharide in combination with a therapeutically
effective amount of parathyroid hormone in a pharmaceutically
acceptable carrier.
[0027] In one aspect, the invention provides a pharmaceutical
composition having a suitable nontoxic, nonionic alkyl glycoside
having a hydrophobic alkyl group joined by a linkage to a
hydrophilic saccharide in combination with a therapeutically
effective amount of a peptide having a molecular weight of about
1-75 kD in a pharmaceutically acceptable carrier, with the proviso
that the peptide is not insulin, calcitonin, and glucagon.
[0028] In one aspect, the invention provides a pharmaceutical
composition having a suitable nontoxic, nonionic alkyl glycoside
having a hydrophobic alkyl group joined by a linkage to a
hydrophilic saccharide in combination with a therapeutically
effective amount erythropoietin in a pharmaceutically acceptable
carrier.
[0029] In one aspect, the invention provides a pharmaceutical
composition having a therapeutically effective amount of an
oligonucleotide in combination with an absorption increasing amount
of an alkylglycoside. The oligonucleotide can be an antisense
oligonucleotide or interfering RNA molecules, such as siRNA or
RNAi. The oligonucleotide typically has a molecular weight of about
1-20 kD and is from about 1-100, 1-50, 1-30, 1-25 or 15-25
nucleotides in length. In another aspect, the oligonucleotide has a
molecular weight of about 5-10 kD. In one aspect, the
alkylglycoside is tetradecyl-beta-D-maltoside.
[0030] In yet another aspect, the invention provides a method of
increasing the bioavailability of a low molecular weight
oligonucleotide in a subject by administering the compound with an
absorption increasing amount of an alkylglycoside, thereby
increasing the bioavailability of the compound in the subject. In
one aspect, the alkylglycoside is tetradecyl-beta-D-maltoside.
[0031] In one aspect, the invention provides a method of increasing
absorption of a compound into the CSF of a subject having
administered intranasally the compound and an absorption increasing
amount of a suitable nontoxic, nonionic alkyl glycoside having a
hydrophobic alkyl group joined by a linkage to a hydrophilic
saccharide.
[0032] In yet another aspect, the invention provides a
pharmaceutical composition having a suitable nontoxic, nonionic
alkyl glycoside having a hydrophobic alkyl group joined by a
linkage to a hydrophilic saccharide in combination with a mucosal
delivery-enhancing agent selected from:
[0033] (a) an aggregation inhibitory agent;
[0034] (b) a charge-modifying agent;
[0035] (c) a pH control agent;
[0036] (d) a degradative enzyme inhibitory agent;
[0037] (e) a mucolytic or mucus clearing agent;
[0038] (f) a ciliostatic agent;
[0039] (g) a membrane penetration-enhancing agent selected from:
[0040] (i) a surfactant; (ii) a bile salt; (ii) a phospholipid
additive, mixed micelle, liposome, or carrier; (iii) an alcohol;
(iv) an enamine; (v) an NO donor compound; (vi) a long-chain
amphipathic molecule; (vii) a small hydrophobic penetration
enhancer; (viii) sodium or a salicylic acid derivative; (ix) a
glycerol ester of acetoacetic acid; (x) a cyclodextrin or
beta-cyclodextrin derivative; (xi) a medium-chain fatty acid; (xii)
a chelating agent; (xiii) an amino acid or salt thereof; (xiv) an
N-acetylamino acid or salt thereof; (xv) an enzyme degradative to a
selected membrane component; (ix) an inhibitor of fatty acid
synthesis; (x) an inhibitor of cholesterol synthesis; and (xi) any
combination of the membrane penetration enhancing agents recited in
(i)-(x);
[0041] (h) a modulatory agent of epithelial junction
physiology;
[0042] (i) a vasodilator agent;
[0043] (j) a selective transport-enhancing agent; and
[0044] (k) a stabilizing delivery vehicle, carrier, mucoadhesive,
support or complex-forming species with which the compound is
effectively combined, associated, contained, encapsulated or bound
resulting in stabilization of the compound for enhanced nasal
mucosal delivery, wherein the formulation of the compound with the
intranasal delivery-enhancing agents provides for increased
bioavailability of the compound in a blood plasma of a subject.
[0045] In one aspect, the invention provides a method of increasing
absorption of a low molecular weight compound into the circulatory
system of a subject by administering, via the oral, ocular, nasal,
nasolacrimal, inhalation or pulmonary, oral cavity (sublingual or
Buccal cell) or CSF delivery route (a) the compound; (b) an
absorption increasing amount of a suitable nontoxic, nonionic alkyl
glycoside having a hydrophobic alkyl group joined by a linkage to a
hydrophilic saccharide; and (c) a mucosal delivery-enhancing
agent.
[0046] In one aspect, the invention provides a method of
controlling caloric intake by administering a composition having a
therapeutic effective amount of exendin-4, or related GLP-1
peptide, with an effective amount of Intravail alkyl
saccharide.
[0047] In another aspect, the invention provides a method of
controlling blood glucose levels in a subject by administering to a
subject a composition comprising a therapeutic effective amount of
exendin-4, or related GLP-1 peptide, with an effective amount of
Intravail alkyl saccharide.
[0048] Still, in another aspect, the invention provides a
controlled release dosage composition comprising:
[0049] (a) a core comprising: [0050] (i) at least one therapeutic
agent or drug; [0051] (ii) at least one alkyl glycoside and/or
saccharide alkyl ester; and
[0052] (b) at least one membrane coating surrounding the core,
wherein the coating is impermeable, permeable, semi-permeable or
porous and becomes more permeable upon sustained contact with
contents of the gastrointestinal tract.
[0053] In another embodiment, the invention provides a method of
administering an alkylglycoside composition by administering a
therapeutically effective amount of at least one alkyglycoside
having an alkyl chain length from about 12 to about 14 carbon
atoms, at least one saccharide with an antibacterial activity, and
at least one therapeutic agent.
[0054] Still in another embodiment, the invention provides a
composition having at least one drug selected from the group
consisting of insulin, PYY, Exendin-4 or other GLP-1 related
peptide, human growth hormone, calcitonin, parathyroid hormone,
truncated parathyroid hormone peptides such as PTH 1-34, EPO,
interferon alpha, interferon beta, interferon gamma, and GCSF and
at least one alkyl saccharide having antibacterial activity.
[0055] In one aspect, the invention provides an antibacterial alkyl
saccharide composition, which includes
n-Dodecyl-4-O-.alpha.-D-glucopyranosyl-.beta.-D-glucopyranoside or
n-tetradecyl-4-O-.alpha.-D-glucopyranosyl-.beta.-D-glucopyranoside.
[0056] Yet, in another aspect, the invention provides an aqueous
drug composition for transmucocal or transdermal administration
having at least one drug and at least one antibacterial agent in a
concentration from about 0.05% to about 0.5%.
[0057] In another aspect, the invention provides a fast-dispersing
drug formulation containing a matrix material and an
alkylsaccharide. The formulation may have a Tmax substantially less
than, and a first-pass effect substantially less than that observed
for an equivalent formulation not containing an alkylsaccharide. In
one embodiment, the formulation may contain about 0.1% to 10%
alkylsaccharide, and exhibits a Tmax substantially less than six
hours and a first-pass effect of less than 40%. The alkylglycoside
may be any suitable alykylglycoside and in a preferred aspect is
dodecyl maltoside, tetradecyl maltoside, sucrose dodecanoate, or
sucrose mono- and di-stearate. The formulation may include a
variety of different therapeutics, such as but not limited to
melatonin, raloxifene, olanzapene and diphenhydramine.
[0058] In another aspect, the invention provides a method for
providing an extended absorption curve by attenuating the
alkylsaccharide concentration in drug formulation to balance
gastric and buccal delivery. For example, this is performed by
providing a drug formulation including a matrix material and an
alkylsaccharide having a Tmax substantially less than, and a
first-pass effect substantially less than that observed for an
equivalent formulation not containing an alkylsaccharide.
[0059] In one aspect, the invention provides a pharmaceutical
composition having a therapeutically effective amount of a
bisphosphonate analog or a triptan analog in combination with an
absorption increasing amount of an alkylglycoside. In various
embodiments, the bisphosphonate analog may be etidronate,
clodronate, tiludronate, pamidronate, neridronate, olpadronate,
alendronate, ibandronate, risedronate, zoledronate, and/or
pharmaceutically acceptable analogs thereof. In an exemplary
embodiment, the bisphosphonate analog is alendronate or
pharmaceutically acceptable analog thereof. In various embodiments,
the triptan analog may be sumatriptan, rizatriptan, naratriptan,
zolmitriptan, eletriptan, almotriptan, frovatriptan and/or
pharmaceutically acceptable analogs thereof. In an exemplary
embodiment, the triptan analog is sumatriptan or pharmaceutically
acceptable analog thereof. In various embodiments, the
alkylglycoside is tetradecyl-beta-D-maltoside.
[0060] In yet another aspect, the invention provides a method of
increasing the bioavailability of a bisphosphonate analog or a
triptan analog in a subject by administering the compound with an
absorption increasing amount of an alkylglycoside, thereby
increasing the bioavailability of the compound in the subject.
[0061] In still another aspect, the invention provides a
composition including a peptide, wherein the peptide includes a
D-amino acid or a site for cyclization, or combination thereof, and
at least one alkylsaccharide, wherein the alkylsaccharide provides
increased enteral absorption of the peptide.
[0062] In yet another aspect, the invention provides method of
increasing enteral adsorption of a peptide in a biphasic manner.
The method includes orally or nasally administering to a subject a
composition comprising at least one peptide, wherein the peptide
comprises a D-amino acid or a site for cyclization, or combination
thereof, and at least one alkylsaccharide, wherein the enteral
absorption of the peptide is increased and systemic serum levels of
the peptide are increased in a biphasic manner.
[0063] In yet another aspect, the invention provides a method of
increasing the bioavailability of a glucagon-like peptide-1 (GLP-1)
analog in a subject. The method includes administering the analog
with an absorption increasing amount of an alkylglycoside, thereby
increasing the bioavailability of the analog in the subject.
[0064] In yet another aspect, the invention provides a
pharmaceutical composition including a glucagon-like peptide-1
(GLP-1) analog; and an absorption increasing amount of an
alkylglycoside.
BRIEF DESCRIPTION OF THE DRAWINGS
[0065] FIG. 1 is a graph showing the intranasal percent
bioavailability compared to intravenous injection and the
subject-to-subject coefficients of variation for MIACALCIN.RTM.
(salmon calcitonin) with and without alkyl glycoside.
[0066] FIG. 2 is a graph showing the effect of intranasal
administration of insulin/0.25% TDM (filled circles) and intranasal
administration of insulin alone (open circles) in reducing blood
glucose levels.
[0067] FIG. 3 is a graph showing the effect of intranasal (closed
triangles) and intraperitoneal (IP) injection (closed circles)
administration of exendin-4/0.25% TDM and IP injection of saline
alone, minus TDM (open circles) in reducing blood glucose levels
following intraperitoneal (IP) injection of glucose (i.e., in a
so-called "glucose tolerance test").
[0068] FIG. 4 is a graph showing the uptake of 1 mg mouse
p-Leu-4]OB3 in 0.3% alkylglycoside tetradecyl-beta-D-maltoside
(Intravail.TM. A3) by male Swiss Webster Mice following
administration by gavage.
[0069] FIG. 5 is a graph showing the uptake of sumatriptan in 0.5%
alkylglycoside tetradecyl-beta-D-maltoside (Intravail.TM. A3) by
canines for both oral and rectal administration.
[0070] FIG. 6 is a graph showing the uptake profile of 30 .mu.g
octreotide in sodium acetate buffer after subcutaneous delivery to
male Swiss Webster mice.
[0071] FIG. 7 is a graph showing the uptake profile of 30 .mu.g
octreotide in 0.5% Intravail.TM. after oral delivery to male Swiss
Webster mice.
[0072] FIG. 8 is a graph showing the uptake profile of 30 .mu.g
octreotide in 1.5% Intravail.TM. after oral delivery to male Swiss
Webster mice.
[0073] FIG. 9 is a graph showing the uptake profile of 30 .mu.g
octreotide in 3.0% Intravail.TM. after oral delivery to male Swiss
Webster mice.
[0074] FIG. 10 is a graph showing blood glucose levels after oral
administration of an alkylglycoside composition including
liraglutide and challenge with glucose.
[0075] FIG. 11 is a graph displaying a dose response curve.
[0076] FIG. 12 is a graph showing the percent of PE in the plasma
(canine model) over time when delivered with alkylglycoside,
n-dodecyl-beta-D-maltoside.
[0077] FIG. 13 is graph showing the percent of PE in the plasma
(canine model) over time when delivered with the alkylglycoside,
sucrose monododecanoate.
DETAILED DESCRIPTION OF THE INVENTION
[0078] The present invention may be understood more readily by
reference to the following detailed description of specific
embodiments and the Examples included therein.
[0079] The present invention is based on the discovery that
therapeutic compositions comprising of least one drug and at least
one surfactant, wherein the surfactant is comprised of at least one
alkyl glycoside and/or at least one saccharide alkyl ester are
stable, non-toxic, non-irritating, anti-bacterial compositions that
increase bioavailability of the drug and have no observable adverse
effects when administered to a subject.
[0080] A "therapeutic composition" can consist of an admixture with
an organic or inorganic carrier or excipient, and can be
compounded, for example, with the usual non toxic, pharmaceutically
acceptable carriers for tablets, pellets, capsules, suppositories,
solutions, emulsions, suspensions, or other form suitable for use.
The carriers, in addition to those disclosed above, can include
glucose, lactose, mannose, gum acacia, gelatin, mannitol, starch
paste, magnesium trisilicate, talc, corn starch, keratin, colloidal
silica, potato starch, urea, medium chain length triglycerides,
dextrans, and other carriers suitable for use in manufacturing
preparations, in solid, semisolid, or liquid form. In addition,
auxiliary stabilizing, thickening or coloring agents can be used,
for example a stabilizing dry agent such as triulose.
[0081] A "drug" is any therapeutic compound, or molecule, or
therapeutic agent, or biologically active compound, including but
not limited to nucleic acids, small molecules, proteins,
polypeptides or peptides and the like.
[0082] The term "nucleic acids" or "oligonucleotide" also denotes
DNA, cDNA, RNA, siRNA, RNAi, dsRNA and the like, which encode
translated and untranslated regions or inhibits translated or
untranslated regions of structural genes encoding a peptide or
protein or regulatory region. For example, a nucleic acid of the
invention can include 5' and 3' untranslated regulatory nucleotide
sequences as well as translated sequences associated with a
structural gene. The term "nucleic acids" or "oligonucleotide" or
grammatical equivalents as used herein, refers to at least two
nucleotides covalently linked together.
[0083] Additionally, the term "oligonucleotide" refers to
structures including modified portions such as modified sugar
moieties, modified base moieties or modified sugar linking
moieties. These modified portions function in a manner similar to
natural bases, natural sugars and natural phosphodiester linkages.
Accordingly, oligonucleotides may have altered base moieties,
altered sugar moieties or altered inter-sugar linkages. Modified
linkages may be, for example, phosphoramide, phosphorothioate,
phosphorodithioate, methyl phosphonate, phosphotriester,
phosphoramidate, O-methylphophoroamidite linkages, or peptide
nucleic acid backbones and linkages. Other analogs may include
oligonucleotides with positive backbones, non-ionic backbones and
non-ribose backbones. The nucleic acid may be DNA, both genomic and
cDNA, RNA or a hybrid, where the nucleic acid contains any
combination of deoxyribo- and ribo-nucleotides, and any combination
of natural or modified bases, including uracil, adenine, thymine,
cytosine, guanine, inosine, xathanine, hypoxathanine, isocytosine,
isoguanine, halogentated bases and the like. Other modifications
may include, for example, deaza or aza purines and pyrimidines used
in place of natural purine and pyrimidine bases; pyrimidine bases
having substituent groups at the 5- or 6-positions, purine bases
having altered or replacement substituent groups at the 2-, 6- or
8-positions, or sugars having substituent groups at their
2'-position, substitutions for one or more of the hydrogen atoms of
the sugar, or carbocyclic or acyclic sugars.
[0084] The term "antisense," as used herein, refers to any
composition containing a nucleic acid sequence which is
complementary to a specific nucleic acid sequence. The term
"antisense strand" is used in reference to a nucleic acid strand
that is complementary to the "sense" strand. Antisense molecules
may be produced by any method including synthesis or transcription.
Once introduced into a cell, the complementary nucleotides combine
with natural sequences produced by the cell to form duplexes and to
block either transcription or translation.
[0085] Antisense molecules include oligonucleotides comprising a
singe-stranded nucleic acid sequence (either RNA or DNA) capable of
binding to target receptor or ligand mRNA (sense) or DNA
(antisense) sequences. The ability to derive an antisense or a
sense oligonucleotide, based upon a cDNA sequence encoding a given
protein. Antisense or sense oligonucleotides further comprise
oligonucleotides having modified sugar-phosphodiester backbones and
wherein such sugar linkages are resistant to endogenous nucleases.
Such oligonucleotides with resistant sugar linkages are stable in
vivo (i.e., capable of resisting enzymatic degradation) but retain
sequence specificity to be able to bind to target nucleotide
sequences.
[0086] RNAi is a phenomenon in which the introduction of dsRNA into
a diverse range of organisms and cell types causes degradation of
the complementary mRNA. In the cell, long dsRNAs are cleaved into
short (e.g., 21-25 nucleotide) small interfering RNAs (siRNAs), by
a ribonuclease. The siRNAs subsequently assemble with protein
components into an RNA-induced silencing complex (RISC), unwinding
in the process. The activated RISC then binds to complementary
transcripts by base pairing interactions between the siRNA
antisense strand and the mRNA. The bound mRNA is then cleaved and
sequence specific degradation of mRNA results in gene silencing. As
used herein, "silencing" refers to a mechanism by which cells shut
down large sections of chromosomal DNA resulting in suppressing the
expression of a particular gene. The RNAi machinery appears to have
evolved to protect the genome from endogenous transposable elements
and from viral infections. Thus, RNAi can be induced by introducing
nucleic acid molecules complementary to the target mRNA to be
degraded.
[0087] Other examples of sense or antisense oligonucleotides
include those oligonucleotides which are covalently linked to
organic moieties and other moieties that increase affinity of the
oligonucleotide for a target nucleic acid sequence, such as
poly-(L-lysine). Further still, intercalating agents, such as
ellipticine, and alkylating agents or metal complexes may be
attached to sense or antisense oligonucleotides to modify binding
specificities of the antisense or sense oligonucleotide for the
target nucleotide sequence.
[0088] A peptide of the invention may be any medically or
diagnostically useful peptide or protein of small to medium size
(i.e., up to about 15 kD, 30 kD, 40 kD, 50 kD, 60 kD, 70 kD, 80 kD,
90 kD, 100 kD, for example). The mechanisms of improved polypeptide
absorption are described in U.S. Pat. No. 5,661,130 which is hereby
incorporated by reference in its entirety. Invention compositions
can be mixed with all such peptides, although the degree to which
the peptide benefits are improved may vary according to the
molecular weight and the physical and chemical properties of the
peptide, and the particular surfactant used. Examples of
polypeptides include vasopressin, vasopressin polypeptide analogs,
desmopressin, glucagon, corticotropin (ACTH), gonadotropin,
calcitonin, C-peptide of insulin, parathyroid hormone (PTH), growth
hormone (HG), human growth hormone (hGH), growth hormone releasing
hormone (GHRH), oxytocin, corticotropin releasing hormone (CRH),
somatostatin or somatostatin polypeptide analogs, gonadotropin
agonist or gonadotrophin agonist polypeptide analogs, human atrial
natriuretic peptide (ANP), human thyroxine releasing hormone (TRH),
follicle stimulating hormone (FSH), prolactin, insulin, insulin
like growth factor-I (IGF-I) somatomedin-C(SM-C), calcitonin,
leptin and the leptin derived short peptide OB-3, melatonin, GLP-1
or Glucagon-like peptide-1 and analogs thereof, such as exenatide,
albiglutide, taspoglutide, liraglutide and lixisenatide, GiP,
neuropeptide pituitary adenylate cyclase, GM-1 ganglioside, nerve
growth factor (NGF), nafarelin, D-tryp6)-LHRH, FGF, VEGF
antagonists, leuprolide, interferon (e.g., .alpha.,.beta., .gamma.)
low molecular weight heparin, PYY, LHRH antagonists, Keratinocyte
Growth Factor (KGF), Glial-Derived Neurotrophic Factor (GDNF),
ghrelin, and ghrelin antagonists. Further, in some aspects, the
peptide or protein is selected from a growth factor, interleukin,
polypeptide vaccine, enzyme, endorphin, glycoprotein, lipoprotein,
or a polypeptide involved in the blood coagulation cascade.
[0089] Certain short peptides composed of approximately 8 to 10
D-amino acids designated Allosteramers.RTM. produced by Allostera
Pharma Inc., Quebec, Canada, have been shown to have an increased
degree of oral bioavailability as well as extended length of time
in the blood stream. Such D-amino acid-containing peptides are
particularly well suited for use with the present invention.
Cyclization, as in cyclic PTH 1-31 (Nemeth 2008), provides another
way to reduce gastrointestinal hydrolysis. Thus, in various
aspects, short peptides containing non-naturally occurring
structural modifications or amino acids are best suited to the
present invention. Peptides comprising less than about 60, 50, 40,
30, 20, 15 or 10 amino acids are contemplated.
[0090] Another example of a peptide containing D-amino acids is the
D-Leu OB-3 peptide, which is orally active when administered in
combination with alkylglycosides, such as
n-dodecyl-beta-D-maltoside.
[0091] Another peptide for use with the present invention is
octreotide acetate (Sandostatin.RTM.). Octreotide is a cyclic
octapeptide used for administration by deep subcutaneous (intrafat)
or intravenous injection for treatment of acromegaly, metastatic
carcinoid tumors where it suppresses or inhibits the severe
diarrhea and flushing episodes associated with the disease, and the
treatment of the profuse watery diarrhea associated with
VIP-secreting tumors. Octreotide acetate is known chemically as
L-Cysteinamide,
D-phenylalanyl-L-cysteinyl-L-phenylalanyl-D-tryptophyl-L-lysyl-L-threonyl-
-N-[2-hydroxy-1-(hydroxymethyl)propyl]-, cyclic
(2.fwdarw.7)-disulfide; [R-(R*, R*)] acetate salt. It is a
long-acting octapeptide with pharmacologic actions mimicking those
of the natural hormone somatostatin and it contains both D-amino
acids as well as cyclization, two properties that stabilize the
molecule against destruction in the gastrointestinal tract.
Octreotide is currently only administered by injection, however as
discussed herein, may be successfully delivered nasally or orally.
Analogs of somatostatin having altered amino acyl sequences have
also been prepared and are suitable for use with the present
invention.
[0092] In one aspect, the present invention provides oral
administration of octreotide or octreotide analogs, such as but not
limited to pentetreotide Dicarba-Analog of Octreotide, or 1-123
Tyr3-octreotide with high bioavailability, circumventing the need
and inconvenience of multiple daily and monthly injections and
preventing needle stick injuries and associated infections of
healthcare providers and family members.
[0093] Other drugs or therapeutic compounds, molecules and/or
agents include cyclic peptides, such as oxytocin, carbetocin, and
demoxytocin, compounds or molecules of the central nervous system
affecting neurotransmitters or neural ion channels (i.e.,
antidepressants (bupropion)), selective serotonin 2c receptor
agonists, anti-seizure agents (topiramate, zonisamide), some
dopamine antagonists, and cannabinoid-1 receptor antagonists
(rimonabant)); leptin/insulin/central nervous system pathway agents
(i.e., leptin analogues, leptin transport and/or leptin receptor
promoters, ciliary neurotrophic factor (Axokine), neuropeptide Y
and agouti-related peptide antagonists, proopiomelanocortin,
cocaine and amphetamine regulated transcript promoters,
alpha-melanocyte-stimulating hormone analogues, melanocortin-4
receptor agonists, protein-tyro sine phosphatase-1B inhibitors,
peroxisome proliferator activated receptor-gamma receptor
antagonists, short-acting bromocriptine (ergo set), somatostatin
agonists (octreotide), and adiponectin); gastrointestinal-neural
pathway agents (i.e., agents that increase glucagon-like peptide-1
activity, such as exenatide (extendin-4), liraglutide,
taspoglutide, albiglutide, lixisenatide and dipeptidyl peptidase IV
inhibitors, protein YY3-36, ghrelin, ghrelin antagonists, amylin
analogues (pramlintide)); and compounds or molecules that may
increase resting metabolic rate "selective" beta-3
stimulators/agonist, melanin concentrating hormone antagonists,
phytostanol analogues, functional oils, P57, amylase inhibitors,
growth hormone fragments, synthetic analogues of
dehydroepiandrosterone sulfate, antagonists of adipocyte
11B-hydroxysteroid dehydrogenase type 1 activity,
corticotropin-releasing hormone agonists, inhibitors of fatty acid
synthesis, carboxypeptidase inhibitors, gastrointestinal lipase
inhibitors (ATL962), melatonin, raloxifene, olanzapene and
diphenhydramine.
[0094] Other drugs or therapeutic compounds include osteoporosis
drugs, such as bisphosphonate analogs. Bisphosphonate analogs, also
known as diphosphonates, are used clinically for the treatment of
conditions such as osteoporosis, osteitis deformans (Paget's
disease of the bone), bone metastasis (with or without
hypercalcaemia), multiple myeloma, osteogenesis imperfecta and
other conditions that feature bone fragility. The class of drugs
inhibit osteoclast action and the resorption of bone. Examples of
bisphosphonates to be admixed with alkylsaccharides for use in the
compositions as described herein include both non-N-containing and
N-containing bisphosphonate analogs. Example of non-N-containing
bisphosphonates include etidronate (Didronel.TM.), clodronate
(Bonefos.TM., Loron.TM.), tiludronate (Skelid.TM.), and
pharmaceutically acceptable analogs thereof. Examples of
N-containing bisphosphonates include pamidronate (Aredia.TM.),
neridronate, olpadronate, alendronate (Fosamax.TM. or
Fosamax+D.TM.), ibandronate (Boniva.TM.), risedronate
(Actonel.TM.), and zoledronate (Zometa.TM. or Reclast.TM.), and
pharmaceutically acceptable analogs thereof.
[0095] Other drugs or therapeutic compounds include drugs, such as
triptan analogs. Triptan analogs are generally a family of
tryptamine based drugs used for the treatment of migraines and
headaches. Their action is attributed to their binding to serotonin
receptors in nerve ending and in cranial blood vessels (causing
their constriction) and subsequent inhibition of pro-inflammatory
neuropeptide release. Examples of triptans to be admixed with
alkylsaccharides for use in the compositions as described herein
include sumatriptan (Imitrex.TM. and Imigran.TM.), rizatriptan
(Maxalt.TM.), naratriptan (Amerge.TM. and Naramig.TM.),
zolmitriptan (Zomig.TM.), eletriptan (Relpax.TM.), almotriptan
(Axert.TM. and Almogran.TM.), frovatriptan (Frova.TM. and
Migard.TM.), and pharmaceutically acceptable analogs thereof.
[0096] The therapeutic composition of the invention includes a drug
and a drug absorption enhancing agent, for example, a surfactant.
The term "surfactant" is any surface active agent that modifies
interfacial tension of water. Typically, surfactants have one
lipophilic and one hydrophilic group in the molecule. Broadly, the
group includes soaps, detergents, emulsifiers, dispersing and
wetting agents, and several groups of antiseptics. More
specifically, surfactants include stearyltriethanolamine, sodium
lauryl sulfate, laurylaminopropionic acid, lecithin, benzalkonium
chloride, benzethonium chloride and glycerin monostearate; and
hydrophilic polymers such as polyvinyl alcohol,
polyvinylpyrrolidone, carboxymethylcellulose sodium,
methylcellulose, hydroxymethylcellulose, hydroxyethylcellulose and
hydroxypropylcellulose.
[0097] Preferably, the surfactant of the invention consists of at
least one suitable alkyl glycoside. As used herein, "alkyl
glycoside" refers to any sugar joined by a linkage to any
hydrophobic alkyl, as is known in the art. Any "suitable" alkyl
glycoside means one that fulfills the limiting characteristics of
the invention, i.e., that the alkyl glycoside be nontoxic and
nonionic, and that it increases the absorption of a compound when
it is administered with the compound via the ocular, nasal,
nasolacrimal, inhalation or pulmonary, oral cavity (sublingual or
Buccal cell), or CSF delivery route. Suitable compounds can be
determined using the methods set forth herein.
[0098] Alkyl glycosides of the invention can be synthesized by
known procedures, i.e., chemically, as described, e.g., in Rosevear
et al., Biochemistry 19:4108-4115 (1980) or Koeltzow and Urfer, J.
Am. Oil Chem. Soc., 61:1651-1655 (1984), U.S. Pat. Nos. 3,219,656
and 3,839,318 or enzymatically, as described, e.g., in Li et al.,
J. Biol. Chem., 266:10723-10726 (1991) or Gopalan et al., J. Biol.
Chem. 267:9629-9638 (1992).
[0099] Alkyl glycosides of the present invention can include, but
are not limited to: alkyl glycosides, such as octyl-, nonyl-,
decyl-, undecyl-, dodecyl-, tridecyl-, tetradecyl-, pentadecyl-,
hexadecyl-, heptadecyl-, and octadecyl-.alpha.- or
.beta.-D-maltoside, -glucoside or -sucroside (synthesized according
to Koeltzow and Urfer; Anatrace Inc., Maumee, Ohio; Calbiochem, San
Diego, Calif.; Fluka Chemie, Switzerland); alkyl thiomaltosides,
such as heptyl, octyl, dodecyl-, tridecyl-, and
tetradecyl-.beta.-D-thiomaltoside (synthesized according to Defaye,
J. and Pederson, C., "Hydrogen Fluoride, Solvent and Reagent for
Carbohydrate Conversion Technology" in Carbohydrates as Organic Raw
Materials, 247-265 (F. W. Lichtenthaler, ed.) VCH Publishers, New
York (1991); Ferenci, T., J. Bacteriol, 144:7-11 (1980)); alkyl
thioglucosides, such as heptyl- or octyl 1-thio .alpha.- or
.beta.-D-glucopyranoside (Anatrace, Inc., Maumee, Ohio; see Saito,
S. and Tsuchiya, T. Chem. Pharm. Bull. 33:503-508 (1985)); alkyl
thiosucroses (synthesized according to, for example, Binder, T. P.
and Robyt, J. F., Carbohydr. Res. 140:9-20 (1985)); alkyl
maltotriosides (synthesized according to Koeltzow and Urfer); long
chain aliphatic carbonic acid amides of sucrose .beta.-amino-alkyl
ethers; (synthesized according to Austrian Patent 382,381 (1987);
Chem. Abstr., 108:114719 (1988) and Gruber and Greber pp. 95-116);
derivatives of palatinose and isomaltamine linked by amide linkage
to an alkyl chain (synthesized according to Kunz, M.,
"Sucrose-based Hydrophilic Building Blocks as Intermediates for the
Synthesis of Surfactants and Polymers" in Carbohydrates as Organic
Raw Materials, 127-153); derivatives of isomaltamine linked by urea
to an alkyl chain (synthesized according to Kunz); long chain
aliphatic carbonic acid ureides of sucrose .beta.-amino-alkyl
ethers (synthesized according to Gruber and Greber, pp. 95-116);
and long chain aliphatic carbonic acid amides of sucrose
.beta.-amino-alkyl ethers (synthesized according to Austrian Patent
382,381 (1987), Chem. Abstr., 108:114719 (1988) and Gruber and
Greber, pp. 95-116).
[0100] Surfactants of the invention consisting of an alkyl
glycoside and/or a sucrose ester have characteristic
hydrophile-lipophile balance (HLB) numbers, which can be calculated
or determined empirically (Schick, M. J. Nonionic Surfactants, p.
607 (New York: Marcel Dekker, Inc. (1967)). The HLB number is a
direct reflection of the hydrophilic character of the surfactant,
i.e., the larger the HLB number, the more hydrophilic the compound.
HLB numbers can be calculated by the formula: (20 times MW
hydrophilic component)/(MW hydrophobic component+MW hydrophilic
component), where MW=molecular weight (Rosen, M. J., Surfactants
and Interfacial Phenomena, pp. 242-245, John Wiley, New York
(1978)). The HLB number is a direct expression of the hydrophilic
character of the surfactant, i.e., the larger the HLB number, the
more hydrophilic the compound. A preferred surfactant has an HLB
number of from about 10 to 20 and an even more preferred range of
from about 11 to 15.
[0101] As described above, the hydrophobic alkyl can thus be chosen
of any desired size, depending on the hydrophobicity desired and
the hydrophilicity of the saccharide moiety. For example, one
preferred range of alkyl chains is from about 9 to about 24 carbon
atoms. An even more preferred range is from about 9 to about 16 or
about 14 carbon atoms. Similarly, some preferred glycosides include
maltose, sucrose, and glucose linked by glycosidic linkage to an
alkyl chain of 9, 10, 12, 13, 14, 16, 18, 20, 22, or 24 carbon
atoms, e.g., nonyl-, decyl-, dodecyl- and tetradecyl sucroside,
glucoside, and maltoside, etc. These compositions are nontoxic,
since they are degraded to an alcohol and an oligosaccharide, and
amphipathic.
[0102] The surfactants of the invention can also include a
saccharide. As use herein, a "saccharide" is inclusive of
monosaccharides, oligosaccharides or polysaccharides in straight
chain or ring forms, or a combination thereof to form a saccharide
chain. Oligosaccharides are saccharides having two or more
monosaccharide residues. The saccharide can be chosen, for example,
from any currently commercially available saccharide species or can
be synthesized. Some examples of the many possible saccharides to
use include glucose, maltose, maltotriose, maltotetraose, sucrose
and trehalose. Preferable saccharides include maltose, sucrose and
glucose.
[0103] The surfactants of the invention can likewise consist of a
sucrose ester. As used herein, "sucrose esters" are sucrose esters
of fatty acids and is a complex of sucrose and fatty acid. Sucrose
esters can take many forms because of the eight hydroxyl groups in
sucrose available for reaction and the many fatty acid groups, from
acetate on up to larger, more bulky fatty acids that can be reacted
with sucrose. This flexibility means that many products and
functionalities can be tailored, based on the fatty acid moiety
used. Sucrose esters have food and non-food uses, especially as
surfactants and emulsifiers, with growing applications in
pharmaceuticals, cosmetics, detergents and food additives. They are
biodegradable, non-toxic and mild to the skin.
[0104] The surfactants of the invention have a hydrophobic alkyl
group linked to a hydrophilic saccharide. The linkage between the
hydrophobic alkyl group and the hydrophilic saccharide can include,
among other possibilities, a glycosidic, thioglycosidic (Horton),
amide (Carbohydrates as Organic Raw Materials, F. W. Lichtenthaler
ed., VCH Publishers, New York, 1991), ureide (Austrian Pat. 386,414
(1988); Chem. Abstr. 110:137536p (1989); see Gruber, H. and Greber,
G., "Reactive Sucrose Derivatives" in Carbohydrates as Organic Raw
Materials, pp. 95-116) or ester linkage (Sugar Esters: Preparation
and Application, J. C. Colbert ed., (Noyes Data Corp., New Jersey),
(1974)). Further, preferred glycosides can include maltose,
sucrose, and glucose linked by glycosidic linkage to an alkyl chain
of about 9-16 carbon atoms, e.g., nonyl-, decyl-, dodecyl- and
tetradecyl sucroside, glucoside, and maltoside. Again, these
compositions are amphipathic and nontoxic, because they degrade to
an alcohol and an oligosaccharide.
[0105] The above examples are illustrative of the types of
glycosides to be used in the methods claimed herein, but the list
is not exhaustive. Derivatives of the above compounds which fit the
criteria of the claims should also be considered when choosing a
glycoside. All of the compounds can be screened for efficacy
following the methods taught herein and in the examples.
[0106] The compositions of the present invention can be
administered in a format selected from the group consisting of a
tablet, a capsule, a suppository, a drop, a spray, an aerosol and a
sustained release or delayed burst format. The spray and the
aerosol can be achieved through use of an appropriate dispenser.
The sustained release format can be an ocular insert, erodible
microparticulates, swelling mucoadhesive particulates, pH sensitive
microparticulates, nanoparticles/latex systems, ion-exchange resins
and other polymeric gels and implants (Ocusert, Alza Corp.,
California; Joshi, A., S. Ping and K. J. Himmelstein, Patent
Application WO 91/19481). These systems maintain prolonged drug
contact with the absorptive surface preventing washout and
nonproductive drug loss. The prolonged drug contact is non-toxic to
the skin and mucosal surfaces.
[0107] The surfactant compositions of the invention are stable. For
example, Baudys et al. in U.S. Pat. No. 5,726,154 show that
calcitonin in an aqueous liquid composition comprising SDS (sodium
dodecyl sulfate, a surfactant) and an organic acid is stable for at
least 6 months. Similarly, the surfactant compositions of the
present invention have improved stabilizing characteristics when
admixed with a drug. No organic acid is required in these
formulations. For example, the composition of the invention
maintains the stability of proteins and peptide therapeutics for
about 6 months, or more, when maintained at about 4.degree. C. to
25.degree. C.
[0108] The stability of the surfactant compositions are, in part,
due to their high no observable adverse effect level (NOAEL). The
Environmental Protection Agency (EPA) defines the no observable
adverse effect level (NOAEL) as the exposure level at which there
are no statistically or biologically significant increases in the
frequency or severity of adverse effects between the exposed
population and its appropriate control. Hence, the term, "no
observable adverse effect level" (or NOAEL) is the greatest
concentration or amount of a substance, found by experiment or
observation, which causes no detectable adverse alteration of
morphology, functional capacity, growth, development, or life span
of the target organism under defined conditions.
[0109] The Food and Agriculture Organization (FAO) of the United
Nations of the World Health Organization (WHO) has shown that some
alkyl glycosides have very high NOAELs, allowing for increased
consumption of these alkyl glycosides without any adverse effect.
This report can be found on the world wide web at
inchem.org/documents/jecfa/jecmono/v10je11.htm. For example, the
NOAEL for sucrose dodecanoate, a sucrose ester used in food
products, is about 20-30 grams/kilogram/day, e.g., a 70 kilogram
person (about 154 lbs.) can consume about 1400-2100 grams (or about
3 to 4.6 pounds) of sucrose dodecanoate per day without any
observable adverse effect. Typically, an acceptable daily intake
for humans is about 1% of the NOAEL, which translates to about
14-21 grams, or 14 million micrograms to 21 million micrograms, per
day, indefinitely. Definitions of NOAELs and other related
definitions can be found on the world wide web at
epa.gov/OCEPAterms. Thus, although some effects may be produced
with alkyl glycoside levels anticipated in the present invention,
the levels are not considered adverse, or precursors to adverse
effects.
[0110] Accordingly, a subject treated with surfactant compositions
of the invention having at least one alkyl glycoside, e.g.,
tetradecylmaltoside (TDM; or Intravail A), at a concentration of
about 0.125% by weight of alkyl glycoside two times per day, or
three times per day, or more depending on the treatment regimen
consumes about 200 to 300 micrograms per day total of TDM. So, the
effective dose of the TDM is at least 1000.times. fold lower than
(i.e., 1/1000) of the NOAEL, and falls far below 1% of the NOAEL,
which is the acceptable daily intake; or in this case about
1/50,000 of the acceptable daily intake. Stated another way, alkyl
glycosides of the present invention have a high NOAEL, such that
the amount or concentration of alkyl glycosides used in the present
invention do not cause an adverse effect and can be safely consumed
without any adverse effect.
[0111] The surfactant compositions of the invention are also stable
because they are physiologically non-toxic and non-irritants. The
term, "nontoxic" means that the alkyl glycoside molecule has a
sufficiently low toxicity to be suitable for human administration
and consumption. Preferred alkyl glycosides are non-irritating to
the tissues to which they are applied. Any alkyl glycoside used
should be of minimal or no toxicity to the cell, such that it does
not cause damage to the cell. Yet, toxicity for any given alkyl
glycoside may vary with the concentration of alkyl glycoside used.
It is also beneficial if the alkyl glycoside chosen is metabolized
or eliminated by the body and if this metabolism or elimination is
done in a manner that will not be harmfully toxic. The term,
"non-irritant" means that the agent does not cause inflammation
following immediate, prolonged or repeated contact with the skin
surface or mucous membranes.
[0112] Moreover, one embodiment of the surfactant compositions, in
particular, the sucrose esters, serve as anti-bacterial agents. An
agent is an "anti-bacterial" agent or substance if the agent or its
equivalent destroy bacteria, or suppress bacterial growth or
reproduction. The anti-bacterial activity of sucrose esters and
their fatty acids have been reported. Tetsuaki et al. (1997) "Lysis
of Bacillus subtilis cells by glycerol and sucrose esters of fatty
acids," Applied and Environmental Microbiology, 53(3):505-508.
Watanabe et al. (2000) describe that galactose and fructose
laureates are particularly effective carbohydrate monoesters.
Watanabe et al., (2000) "Antibacterial carbohydrate monoesters
suppressing cell growth of Streptococcus mutan in the presence of
sucrose," Curr Microbiol 41(3): 210-213. Hence, the present
invention is not limited to the sucrose ester described herein, but
encompasses other carbohydrate esters, including galactose and
fructose esters, that suppress bacterial growth and
reproduction.
[0113] In general, all useful antimicrobial agents are toxic
substances. See Sutton and Porter (2002), "Development of the
antimicrobial effectiveness test as USP Chapter <51>," 56(6):
300-311, which is incorporated herein by reference in its entirety.
For example, commonly used antimicrobial agents such as
benzalkonium chloride are highly toxic as demonstrated by electron
micrograph studies in which significant disruption of the
mucociliary surfaces are observed at concentrations of benzalkonium
far below what is commonly used in intranasal formulations. See for
example Sebahattin Cureoglu, Murat Akkus, Ustiin Osma, Mehmet
Yaldiz, Faruk Oktay, Belgin Can, Cengiz Guven, Muhammet Tekin, and
Faruk Meric (2002), "The effect of benzalkonium chloride an
electron microscopy study," Eur Arch Otorhinolaryngol
259:362-364.
[0114] The surfactant compositions of the invention are typically
present at a level of from about 0.01% to 20% by weight. More
preferred levels of incorporation are from about 0.01% to 5% by
weight, from about 0.01% to 2% by weight, from about 0.01% to 1%,
most preferably from about 0.01% to 0.125% by weight. The
surfactant is preferably formulated to be compatible with other
components present in the composition. In liquid, or gel, or
capsule, or injectable, or spray compositions the surfactant is
most preferably formulated such that it promotes, or at least does
not degrade, the stability of any protein or enzyme in these
compositions. Further, the invention optimizes the concentration by
keeping the concentration of absorption enhancer as low as
possible, while still maintaining the desired effect.
[0115] The compositions of the invention when administered to the
subject, yield enhanced mucosal delivery of the biologically active
compound(s), or drug, with a peak concentration (or Cmax) of the
compound(s) in a tissue, or fluid, or in a blood plasma of the
subject that is about 15%, 20%, or 50% or greater as compared to a
Cmax of the compound(s) in a tissue (e.g., CNS), or fluid, or blood
plasma following intramuscular injection of an equivalent
concentration of the compound(s) to the subject.
[0116] The measure of how much of the drug or compound(s) reaches
the bloodstream in a set period of time, e.g., 24 hours can also be
calculated by plotting drug blood concentration at various times
during a 24-hour or longer period and then measuring the area under
the curve (AUC) between 0 and 24 hours. Similarly, a measure of
drug efficacy can also be determined from a time to maximal
concentration (tmax) of the biologically active compound(s) in a
tissue (e.g., CNS) or fluid or in the blood plasma of the subject
between about 0.1 to 1.0 hours. The therapeutic compositions of the
invention increase the speed of onset of drug action (i.e., reduce
Tmax) by a factor of about 1.5-fold to 2-fold.
[0117] Also, the therapeutic compositions or formulations of the
invention can be administered or delivered to a subject in need
systemically or locally. Suitable routes may, for example, include
oral, ocular, nasal, nasolacrimal, inhalation or pulmonary, oral
cavity (sublingual or Buccal cell), transmucosal administration,
vaginal, rectal, parenteral delivery, including intramuscular,
subcutaneous, intravenous, intraperitoneal, or CSF delivery.
Moreover, the mode of delivery e.g., liquid, gel, tablet, spray,
etc. will also depend on the method of delivery to the subject.
[0118] Additionally, the therapeutic compositions of the invention
can consist of a pharmaceutically acceptable carrier. A
"pharmaceutically acceptable carrier" is an aqueous or non aqueous
agent, for example alcoholic or oleaginous, or a mixture thereof,
and can contain a surfactant, emollient, lubricant, stabilizer,
dye, perfume, preservative, acid or base for adjustment of pH, a
solvent, emulsifier, gelling agent, moisturizer, stabilizer,
wetting agent, time release agent, humectant, or other component
commonly included in a particular form of pharmaceutical
composition. Pharmaceutically acceptable carriers are well known in
the art and include, for example, aqueous solutions such as water
or physiologically buffered saline or other solvents or vehicles
such as glycols, glycerol, and oils such as olive oil or injectable
organic esters. A pharmaceutically acceptable carrier can contain
physiologically acceptable compounds that act, for example, to
stabilize or to increase the absorption of the specific inhibitor,
for example, carbohydrates, such as glucose, sucrose or dextrans,
antioxidants, such as ascorbic acid or glutathione, chelating
agents, low molecular weight proteins or other stabilizers or
excipients. A pharmaceutically acceptable carrier can also be
selected from substances such as distilled water, benzyl alcohol,
lactose, starches, talc, magnesium stearate, polyvinylpyrrolidone,
alginic acid, colloidal silica, titanium dioxide, and flavoring
agents.
[0119] Additionally, to decrease susceptibility of a peptide drug
to hydrolytic cleavage in compositions containing alkyl saccharides
or saccharide alkyl esters, various oxygen atoms within the drugs
can be substituted for by sulfur (Defaye, J. and Gelas, J. in
Studies in Natural Product Chemistry (Atta-ur-Rahman, ed.) Vol. 8,
pp. 315-357, Elsevier, Amsterdam, 1991). For example, the
heteroatom of the sugar ring can be either oxygen or sulfur, or the
linkage between monosaccharides in an oligosaccharide can be oxygen
or sulfur (Horton, D. and Wander, J. D., "Thio Sugars and
Derivatives," The Carbohydrates: Chemistry and Biochemistry, 2d.
Ed. Vol. B3, (W. Reyman and D. Horton eds.), pp. 799-842, (Academic
Press, New York), (1972)). Oligosaccharides can have either .alpha.
(alpha) or .beta. (beta) anomeric configuration (see Pacsu, E., et
al. in Methods in Carbohydrate Chemistry (R. L. Whistler, et al.,
eds.) Vol. 2, pp. 376-385, Academic Press, New York 1963).
[0120] A composition of the invention can be prepared in tablet
form by mixing a therapeutic agent or drug and one alky glycoside
and/or saccharide alkyl ester according to the invention, and an
appropriate pharmaceutical carrier or excipient, for example
mannitol, corn starch, polyvinylpyrrolidone or the like,
granulating the mixture and finally compressing it in the presence
of a pharmaceutical carrier such as corn starch, magnesium stearate
or the like. If necessary, the formulation thus prepared may
include a sugar-coating or enteric coating or covered in such a way
that the active principle is released gradually, for example, in
the appropriate pH medium.
[0121] The term "enteric coating," is a polymer encasing,
surrounding, or forming a layer, or membrane around the therapeutic
composition or core. Also, the enteric coating can contain a drug
which is compatible or incompatible with the coating. One tablet
composition may include an enteric coating polymer with a
compatible drug which dissolves or releases the drug at higher pH
levels (e.g., pH greater than 4.0, greater than 4.5, greater than
5.0 or higher) and not at low pH levels (e.g., pH 4 or less); or
the reverse.
[0122] In a preferred embodiment, the dose dependent release form
of the invention is a tablet comprising: (a) a core comprising: (i)
a therapeutic agent or drug; (ii) a surfactant comprising at least
one alkyl glycoside and/or saccharide alkyl ester; and (b) at least
one membrane coating surrounding the core, wherein the coating is
an impermeable, permeable, semi-permeable or porous coating and
becomes more permeable or porous upon contacting an aqueous
environment of a defined pH. The term "membrane" is synonymous with
"coating," or equivalents thereof. The terms are used to identify a
region of a medicament, for example, a tablet, that is impermeable,
permeable, semi-permeable or porous to an aqueous solution(s) or
bodily fluid(s), and/or to the therapeutic agent(s) or drug(s)
encapsulated therein. If the membrane is permeable, semi-permeable
or porous to the drug, the drug can be released through the
openings or pores of the membrane in solution or in vivo. The
porous membrane can be manufactured mechanically (e.g., drilling
microscopic holes or pores in the membrane layer using a laser), or
it can be imparted due to the physiochemical properties of the
coating polymer(s). Membrane or coating polymers of the invention
are well known in the art, and include cellulose esters, cellulose
diesters, cellulose triesters, cellulose ethers, cellulose
ester-ether, cellulose acylate, cellulose diacylate, cellulose
triacylate, cellulose acetate, cellulose diacetate, cellulose
triacetate, cellulose acetate propionate, and cellulose acetate
butyrate. Other suitable polymers are described in U.S. Pat. Nos.
3,845,770, 3,916,899, 4,008,719, 4,036,228 and 4,11210 which are
incorporated herein by reference.
[0123] Further, the enteric coating according to the invention can
include a plasticizer, and a sufficient amount of sodium hydroxide
(NaOH) to effect or adjust the pH of the suspension in solution or
in vivo. Examples of plasticizers include triethyl citrate,
triacetin, tributyl sebecate, or polyethylene glycol. Other
alkalizing agents, including potassium hydroxide, calcium
carbonate, sodium carboxymethylcellulose, magnesium oxide, and
magnesium hydroxide can also be used to effect or adjust the pH of
the suspension in solution or in vivo.
[0124] Accordingly, in one embodiment, an enteric coating can be
designed to release a certain percentage of a drug or drugs in
certain mediums with a certain pH or pH range. For example, the
therapeutic composition of the invention may include at least one
enteric coating encasing or protecting at least one drug which is
chemically unstable in an acidic environment (e.g., the stomach).
The enteric coating protects the drug from the acidic environment
(e.g., pH<3), while releasing the drug in locations which are
less acidic, for example, regions of the small and large intestine
where the pH is 3, or 4, or 5, or greater. A medicament of this
nature will travel from one region of the gastrointestinal tract to
the other, for example, it takes about 2 to about 4 hours for a
drug to move from the stomach to the small intestine (duodenum,
jejunum and ileum). During this passage or transit, the pH changes
from about 3 (e.g., stomach) to 4, or 5, or to about a pH of 6 or 7
or greater. Thus, the enteric coating allows the core containing
the drug to remain substantially intact, and prevents premature
drug release or the acid from penetrating and de-stabilizing the
drug.
[0125] Examples of suitable enteric polymers include but are not
limited to cellulose acetate phthalate,
hydroxypropylmethylcellulose phthalate, polyvinylacetate phthalate,
methacrylic acid copolymer, shellac, cellulose acetate
trimellitate, hydroxypropylmethylcellulose acetate succinate,
hydroxypropylmethylcellulose phthalate, cellulose acetate
phthalate, cellulose acetate succinate, cellulose acetate malate,
cellulose benzoate phthalate, cellulose propionate phthalate,
methylcellulose phthalate, carboxymethylethylcellulose,
ethylhydroxyethylcellulose phthalate, shellac, styrene-acrylic acid
copolymer, methyl acrylate-acrylic acid copolymer, methyl
acrylate-methacrylic acid copolymer, butyl acrylate-styrene-acrylic
acid copolymer, methacrylic acid-methyl methacrylate copolymer,
methacrylic acid-ethyl acrylate copolymer, methyl
acrylate-methacrylic acid-octyl acrylate copolymer, vinyl
acetate-maleic acid anhydride copolymer, styrene-maleic acid
anhydride copolymer, styrene-maleic acid monoester copolymer, vinyl
methyl ether-maleic acid anhydride copolymer, ethylene-maleic acid
anhydride copolymer, vinyl butyl ether-maleic acid anhydride
copolymer, acrylonitrile-methyl acrylate-maleic acid anhydride
copolymer, butyl acrylate-styrene-maleic acid anhydride copolymer,
polyvinyl alcohol phthalate, polyvinyl acetal phthalate, polyvinyl
butylate phthalate and polyvinyl acetoacetal phthalate, or
combinations thereof. One skilled in the art will appreciate that
other hydrophilic, hydrophobic and enteric coating polymers may be
readily employed, singly or in any combination, as all or part of a
coating according to the invention.
[0126] The therapeutic compositions of the invention in the form of
a tablet can have a plurality of coatings, for example, a
hydrophilic coating (e.g., hydroxypropylmethylcellulose), and/or a
hydrophobic coating (e.g., alkylcelluloses), and/or an enteric
coating. For example, the tablet core can be encases by a plurality
of the same type of coating, or a plurality of different types of
coating selected from a hydrophilic, hydrophobic or enteric
coating. Hence, it is anticipated that a tablet can be designed
having at least one, but can have more than one layer consisting of
the same or different coatings dependent on the target tissue or
purpose of the drug or drugs. For example the tablet core layer may
have a first composition enclosed by a first coating layer (e.g.,
hydrophilic, hydrophobic, or enteri-coating), and a second same or
different composition or drug having the same or different dosage
can be enclosed in second coating layer, etc. This layering of
various coatings provides for a first, second, third, or more
gradual or dose dependent release of the same or different drug
containing composition.
[0127] In a preferred embodiment, a first dosage of a first
composition of the invention is contained in a tablet core and with
an enteric-coating such that the enteric-coating protects and
prevents the composition contained therein from breaking down or
being released into the stomach. In another example, the first
loading dose of the therapeutic composition is included in the
first layer and consists of from about 10% to about 40% of the
total amount of the total composition included in the formulation
or tablet. In a second loading dose, another percentage of the
total dose of the composition is released. The invention
contemplates as many time release doses as is necessary in a
treatment regimen. Thus, in certain aspects, a single coating or
plurality of coating layers is in an amount ranging from about 2%
to 6% by weight, preferably about 2% to about 5%, even more
preferably from about 2% to about 3% by weight of the coated unit
dosage form.
[0128] Accordingly, the composition preparations of the invention
make it possible for contents of a hard capsule or tablet to be
selectively released at a desired site the more distal parts of the
gastro-intestinal tract (e.g., small and large intestine) by
selecting the a suitable pH-soluble polymer for a specific region.
Mechanical expulsion of the composition preparations may also be
achieved by inclusion of a water absorbing polymer that expands
upon water absorption within a hard semi-permeable capsule thus
expelling composition through an opening in the hard capsule.
[0129] Drugs particularly suited for dose dependent time release
include but are not limited to insulin like growth factor-I
(IGF-I), somatomedin-C(SM-C; diabetes, nerve function, renal
function), insulin (diabetes), calcitonin (osteoporosis), leptin
(obesity; infertility), leptin derived short peptide (OB-3), hGH
(AIDs wasting, dwarfism), human parathyroid hormone (PTH)
(osteoporosis), melatonin (sleep), GLP-1 or Glucagon-like peptide-1
(diabetes), GiP (diabetes), pituitary adenylate cyclase-activating
polypeptide (PACAP) and islet function (diabetes), GM-1
ganglioside, (Alzheimers), nerve growth factor (NGF), (Alzheimers),
nafarelin (endometriosis), Synarel.RTM. (nafarelin acetate nasal
solution), (D-tryp6)-LHRH (fertility), FGF (duodenal ulcer, macular
degeneration, burns, wounds, spinal cord injuries, repair of bone
and cartilage damage), VEGF antagonists (to block the receptor),
VEGF (agonist) neonatal distress syndrome; ALS), leuprolide
(prostate and breast cancer), interferon-alpha (chronic hepatitis
C), low molecular weight heparin (blood clotting, deep vein
thrombosis), PYY (obesity), LHRH antagonists (fertility), LH
(luteinizing hormone), ghrelin antagonists (obesity), KGF
(Parkinson's), GDNF (Parkinsons), G-CSF (erythropoiesis in cancer),
Imitrex (migraine), Integrelin (anticoagulation), Natrecor.RTM.
(congestive heart failure), human B-type natriuretic peptide
(hBNP), SYNAREL.RTM. (Searl; nafarelin acetate nasal solution),
Sandostatin (growth hormone replacement), Forteo (osteoporosis),
DDAVP.RTM. Nasal Spray (desmopressin acetate), Cetrotide.RTM.
(cetrorelix acetate for injection), Antagon.TM. (ganirelix
acetate), Angiomax (bivalirudin; thrombin inhibitor), Accolate.RTM.
(zafirlukast; injectable), Exendin-4 (Exanatide; diabetes),
SYMLIN.RTM. (pramlintide acetate; synthetic amylin; diabetes),
desmopressin, glucagon, ACTH (corticotrophin), C-peptide of
insulin, GHRH and analogs (GnRHa), growth hormone releasing
hormone, oxytocin, corticotropin releasing hormone (CRH), atrial
natriuretic peptide (ANP), thyroxine releasing hormone (TRHrh),
follicle stimulating hormone (FSH), prolactin, tobramycin ocular
(corneal infections), Vasopressin, desmopresin, Fuzeon (Roche; HIV
fusion inhibitor MW 4492), thymalfasin, and Eptifibatide.
[0130] Further, it will be understood by one skilled in the art,
that the specific dose level and frequency of dosage for any
particular subject in need of treatment may be varied and will
depend upon a variety of factors including the activity of the
specific compound employed, the metabolic stability and length of
action of that compound, the age, body weight, general health, sex,
diet, mode and time of administration, rate of excretion, drug
combination, the severity of the particular condition, and the host
undergoing therapy.
[0131] It has been shown that alkyl glycosides, particularly
alkylmaltosides and more specifically, dodecylmalto side (DDM) and
tetradecylmalto side (TDM), stabilize insulin in solution and
prevent aggregation of the peptide. Hovgaard et al., "Insulin
Stabilization and GI absorption," J. Control. Rel., 19 (1992)
458-463, cited in Hovgaard et al., "Stabilization of insulin by
alkylmaltosides: A spectroscopic evaluation," Int. J. Pharmaceutics
132 (1996) 107-113 (hereinafter, "Hovgaard-1"). Further, Hovgaard-1
shows that even after 57 days, the DDM-insulin complex remained
stable and possessed nearly full biological activity. It is
postulated that the stability of the complex is due to the length
of the alkyl group (number of carbon atoms) and the higher ratio of
DDM to insulin ratio the better (e.g., 4:1 and 16:1; see FIG. 1 in
Hovgaard 1). However, according to Hovgaard-1, although the
DDM-insulin complex was stable, the same stability was not shown
for other maltosides. Yet, in a related study, Hovgaard et al.
(1996) demonstrated that when DDM-insulin was orally administered
to animals in vivo, bioavailability of the complex was weak (e.g.,
0.5%-1% bioavailability). Hovgaard et al., "Stabilization of
insulin by alkylmaltoside. B. Oral absorption in vivo in rats,"
Int. J. Pharmaceutics 132 (1996) 115-121 (Hovgaard-2). Hence, an
improved aspect of the invention is that the surfactant increases
the bioavailability of a drug to the target tissues, organs, system
etc., as well as increase drug stability.
[0132] Accordingly, one aspect of the invention is to provide
therapeutic compositions having at least one drug and one
surfactant, wherein the surfactant further consists of at least one
alkyl glycoside and/or saccharide alkyl ester formulation which
enhances the bioavailability of the drug. Determining the
bioavailability of drug formulations is described herein. As used
herein, "bioavailability" is the rate and extent to which the
active substance, or moiety, which reaches the systemic circulation
as an intact drug. The bioavailability of any drug will depend on
how well is adsorbed and how much of it escapes being removed from
the liver.
[0133] To determine absolute bioavailability, the tested drug and
mode of administration is measured against an intravenous reference
dose. The bioavailability of the intravenous dose is 100% by
definition. For example, animals or volunteering humans are given
an intravenous injections and corresponding oral doses of a drug.
Urinary or plasma samples are taken over a period of time and
levels of the drug over that period of time are determined.
[0134] The areas under the curve (AUC), of the plasma drug
concentration versus time curves, are plotted for both the
intravenous and the oral doses, and calculation of the
bioavailability of both formulations is by simple proportion. For
example, if the same intravenous and oral doses are given, and the
oral AUC is 50% of the intravenous AUC, the bioavailability of the
oral formulation is 50%. Note that the bioavailability of any drug
is due to many factors including incomplete absorption, first pass
clearance or a combination of these (discussed more below).
Further, the peak concentration (or C.sub.max) of the plasma drug
concentration is also measured to the peak concentration
(C.sub.max) of the plasma drug concentration following
intramuscular (IM) injection of an equivalent concentration the
drug. Moreover, the time to maximal concentration (or t.sub.max) of
the plasma drug is about 0.1 to 1.0 hours.
[0135] To determine the relative bioavailability of more than one
formulation of a drug (e.g., an alkyl glycoside or saccharide alkyl
ester drug formulation), bioavailability of the formulations are
assessed against each other as one or both drugs could be subject
to first pass clearance (discussed more below) and thus undetected.
For example, a first oral formulation is assessed against a second
oral formulation. The second formulation is used as a reference to
assess the bioavailability of the first. This type of study
provides a measure of the relative performance of two formulations
in getting a drug absorbed.
[0136] Bioavailabilities of drugs are inconsistent and vary greatly
from one drug to the next. For example, the bioavailability of
MIACALCIN.RTM. (salmon calcitonin from Novartis) nasal spray, a
prescription medication for the treatment of postmenopausal
osteoporosis in women, has a mean bioavailability of about 3%
(range is 0.3%-30.6%; see FIG. 1). The MIACALCIN.RTM. product
information sheet can be found on the world wide web at
miacakin.com/info/howWorks/index.jsp and
drugs.com/PDR/Miacalcin_Nasal_Spray.html. The data on
MIACALCIN.RTM., which was obtained by various investigators using
different methods and human subjects, show great variability in the
drug's bioavailability, e.g., in normal volunteers only .about.3%
of the nasally administered dose is bioavailable, as compared to
the same dose administered by intramuscular injection
(MIACALCIN.RTM. product insert). This represents two orders of a
magnitude in variability and is undesirable to the consumer.
[0137] Poor bioavailability of a drug can also be observed in
NASCOBAL.RTM. (Nastech), or cyanocobalamin, which is used for the
treatment and maintenance of the hematologic status of patients who
are in remission following intramuscular vitamin B12 therapies. The
gel formulation was administered intranasally and the
bioavailability of B.sub.12 was compared to intramuscular B12
injections. The peak concentrations of B12 (or the Tmax) was
reached in 1-2 hours after intranasal administration, and relative
to the intramuscular injection, the bioavailability of B12 nasal
gel was found to be about 8.9% (90% confidence intervals, 7.1% to
11.2%).
[0138] The alkyl glycosides or sucrose esters of the present
invention include any compounds now known or later discovered.
Drugs which are particularly well suited for admixture with the
alkyl glycosides and/or saccharide alkyl esters of the invention
are those that are difficult to administer by other methods, e.g.,
drugs that are degraded in the gastrointestinal (GI) tract or those
that are not absorbed well from the GI tract, or drugs that can be
self-administered via the ocular, nasal, nasolacrimal, inhalation,
or CSF delivery route instead of traditional methods such as
injection. Some specific examples include peptides, polypeptides,
proteins, nucleic acids and other macromolecules, for example,
peptide hormones, such as insulin and calcitonin, enkephalins,
glucagon and hypoglycemic agents such as tolbutamide and glyburide,
and agents which are poorly absorbed by enteral routes, such as
griseofulvin, an antifungal agent. Other compounds include, for
example, nicotine, interferon (e.g., alpha, beta, gamma), PYY,
GLP-1, synthetic exendin-4 (Exenatide), parathyroid hormone, and
human growth hormone or other low molecular weight peptides and
proteins.
[0139] Alternatively, bioavailability of a drug can be determined
by measuring the levels of the drug's first pass clearance by the
liver. Alkyl glycosides and/or saccharide alkyl ester compositions
of the invention administered intranasally or via oral cavity
(sublingual or Buccal cell) do not enter the hepatic portal blood
system, thereby avoiding first pass clearance by the liver.
Avoiding first past clearance of these formulations by the liver is
described herein. The term, "first pass liver clearance" is the
extent to which the drug is removed by the liver during its first
passage in the portal blood through the liver to the systemic
circulation. This is also called first pass metabolism or first
pass extraction.
[0140] The two major routes of drug elimination from the body are
excretion by the kidneys whereby the drug is unchanged; and
elimination by the liver, whereby the drug is metabolized. The
balance between these two routes depends on the relative efficiency
of the two processes. The present invention describes herein
elimination by the liver or liver clearance. First pass liver
clearance is described by Birkett et al (1990 and 1991), which is
incorporated by reference in its entirety. Birkett et al., Aust
Prescr, 13(1990):88-9; and Birkett et al., Austra Prescr 14:14-16
(1991).
[0141] Blood carrying drug from the systemic circulation enter the
liver via the portal vein, and the liver in turn extracts a certain
percentage or ratio (i.e., 0.5 or 50%) of that drug. The remainder
left over (i.e., 0.2 or 20%) re-enters the systemic circulation via
the hepatic vein. This rate of clearance of the drug is called the
hepatic extraction ratio. It is the fraction of the drug in the
blood which is irreversibly removed (or extracted) during the first
pass of the blood through the liver. If no drug is extracted, the
hepatic extraction ratio is zero. Conversely, if the drug is highly
extracted in the first pass through the liver, the hepatic
extraction ratio may be as high as 100% or 1.0. In general,
clearance of the drug by the liver depends then on the rate of
delivery of that drug to the liver (or the hepatic blood flow), and
on the efficiency of removal of that drug (or the extraction
ratio).
[0142] Therefore, the net equation used to determine hepatic
clearance is:
(hepatic clearance-blood flow)=(unbound fraction*intrinsic
clearance)/blood flow+(unbound fraction*intrinsic clearance)
(1)
[0143] The "unbound fraction" of drug is dependent on how tightly
the drug is bound to proteins and cells in the blood. In general,
it is only this unbound (or free) drug which is available for
diffusion from the blood into the liver cell. In the absence of
hepatic blood flow and protein binding, the "intrinsic clearance"
is the ability of the liver to remove (or metabolize) that drug. In
biochemical terms, it is a measure of liver enzyme activity for a
particular drug substrate. Again, although intrinsic clearance can
be high, drugs cannot be cleared more rapidly than that presented
to the liver. In simple terms, there are two situations: where
liver enzyme activity is very high or very low (i.e., high
extraction ratio or low extraction ratio).
[0144] When liver enzyme activity is low, the equation simplifies
to:
hepatic clearance=unbound fraction*intrinsic clearance (2)
[0145] Clearance then is independent of blood flow, but instead
depends directly on the degree of protein binding in the blood and
the activity of drug metabolizing enzymes towards that drug.
[0146] In contrast, when liver enzyme activity is high, the
equation is:
hepatic clearance=liver blood flow (3)
[0147] In this scenario, because the enzymes are so active the
liver removes most of the drug presented to it and the extraction
ratio is high. Thus, the only factor determining the actual hepatic
clearance is the rate of supply of drug to the liver (or hepatic
blood flow).
[0148] First pass liver clearance is important because even small
changes in the extraction of drugs can cause large changes in
bioavailability. For example, if the bioavailability of drug A by
oral administration is 20% by the time it reaches the systemic
circulation, and the same drug A by intravenous administration is
100%, absent no other complicating factors, the oral dose will
therefore have to be 5 times the intravenous dose to achieve
similar plasma concentrations.
[0149] Secondly, in some instances where liver enzyme activity is
very high, drug formulations should be designed to have the drug
pass directly through to the systemic circulation and avoid first
pass liver clearance all together. For example, drugs administered
intranasally, sublingual, buccal, rectal, vagina, etc. directly
enter the systemic circulation and do not enter the hepatic portal
blood circulation to be partially or fully extracted by the liver.
Alternatively, where drugs cannot be administered by the above
means, a tablet with at least one enteric-coating layer to prevent
release of the drug in the stomach (i.e., highly acidic
environment) is provided. Thus, an objective of the invention is to
administer drugs using these alternative routes.
[0150] Additionally, first pass liver clearance is an important
factor because many patients are on more than one drug regimen, and
this may cause drug interactions which increase or decrease liver
enzyme activity; thereby increasing or decreasing metabolism
(increasing or decreasing the hepatic extraction ratio) of the drug
of interest.
[0151] Hence, therapeutic compositions of the invention can be
administered directly to the systemic circulatory system and avoid
first pass liver clearance. Avoiding first pass clearance assures
that more of the drug will be available to the system. Stated
another way, by avoiding first pass liver clearance, the
bioavailability of the drug is increased.
[0152] The present invention also relates to methods of increasing
absorption of a low molecular compound into the circulatory system
of a subject comprising administering via the oral, ocular, nasal,
nasolacrimal, inhalation, or the CSF delivery route the compound
and an absorption increasing amount of a suitable nontoxic,
nonionic alkyl glycoside having a hydrophobic alkyl joined by a
linkage to a hydrophilic saccharide.
[0153] The composition formulation is appropriately selected
according to the administration route, such as oral administration
(oral preparation), external administration (e.g., ointment),
injection (preparations for injection), and mucosal administration
(e.g., buccal and suppository) etc. For example, excipients (e.g.,
starch, lactose, crystalline cellulose, calcium lactate, magnesium
aluminometasilicate and anhydrous silicate), disintegrators (e.g.,
carboxymethylcellulose and calcium carboxymethylcellulose),
lubricants (e.g., magnesium stearate and talc), coating agents
(e.g., hydroxyethylcellulose), and flavoring agents can be used for
oral and mucosal formulations; whereas, solubilizers and auxiliary
solubilizers capable of forming aqueous injections (e.g., distilled
water for injection, physiological saline and propylene glycol),
suspending agents (e.g., surfactant such as polysorbate 80), pH
regulators (e.g., organic acid and metal salt thereof) and
stabilizers are used for injections; and aqueous or oily
solubilizers and auxiliary solubilizers (e.g., alcohols and fatty
acid esters), tackifiers (e.g., carboxy vinyl polymer and
polysaccharides) and emulsifiers (e.g., surfactant) are used for
external agents. The drug and the alkyl glycoside can be admixed,
mixed, or blended along with the above excipients, disintegrators,
coating polymers, solubilizers, suspending agents, etc., prior to
administration, or they can be administered sequentially, in either
order. It is preferred that they be mixed prior to
administration.
[0154] The term, "mucosal delivery-enhancing agent" includes agents
which enhance the release or solubility (e.g., from a formulation
delivery vehicle), diffusion rate, penetration capacity and timing,
uptake, residence time, stability, effective half-life, peak or
sustained concentration levels, clearance and other desired mucosal
delivery characteristics (e.g., as measured at the site of
delivery, or at a selected target site of activity such as the
bloodstream or central nervous system) of a compound(s) (e.g.,
biologically active compound). Enhancement of mucosal delivery can
occur by any of a variety of mechanisms, including, for example, by
increasing the diffusion, transport, persistence or stability of
the compound, increasing membrane fluidity, modulating the
availability or action of calcium and other ions that regulate
intracellular or paracellular permeation, solubilizing mucosal
membrane components (e.g., lipids), changing non-protein and
protein sulfhydryl levels in mucosal tissues, increasing water flux
across the mucosal surface, modulating epithelial junction
physiology, reducing the viscosity of mucus overlying the mucosal
epithelium, reducing mucociliary clearance rates, and other
mechanisms.
[0155] Exemplary mucosal delivery enhancing agents include the
following agents and any combinations thereof: [0156] (a) an
aggregation inhibitory agent; [0157] (b) a charge-modifying agent;
[0158] (c) a pH control agent; [0159] (d) a degradative enzyme
inhibitory agent; [0160] (e) a mucolytic or mucus clearing agent;
[0161] (f) a ciliostatic agent; [0162] (g) a membrane
penetration-enhancing agent selected from: [0163] (i) a surfactant;
(ii) a bile salt; (ii) a phospholipid additive, mixed micelle,
liposome, or carrier; (iii) an alcohol; (iv) an enamine; (v) an NO
donor compound; (vi) a long-chain amphipathic molecule; (vii) a
small hydrophobic penetration enhancer; (viii) sodium or a
salicylic acid derivative; (ix) a glycerol ester of acetoacetic
acid; (x) a cyclodextrin or beta-cyclodextrin derivative; (xi) a
medium-chain fatty acid; (xii) a chelating agent; (xiii) an amino
acid or salt thereof; (xiv) an N-acetylamino acid or salt thereof;
(xv) an enzyme degradative to a selected membrane component; (ix)
an inhibitor of fatty acid synthesis; (x) an inhibitor of
cholesterol synthesis; and (xi) any combination of the membrane
penetration enhancing agents recited in (i)-(x); [0164] (h) a
modulatory agent of epithelial junction physiology; [0165] (i) a
vasodilator agent; [0166] (j) a selective transport-enhancing
agent; and [0167] (k) a stabilizing delivery vehicle, carrier,
mucoadhesive, support or complex-forming species with which the
compound is effectively combined, associated, contained,
encapsulated or bound resulting in stabilization of the compound
for enhanced nasal mucosal delivery, wherein the formulation of the
compound with the intranasal delivery-enhancing agents provides for
increased bioavailability of the compound in a blood plasma of a
subject.
[0168] Additional mucosal delivery-enhancing agents include, for
example, citric acid, sodium citrate, propylene glycol, glycerin,
ascorbic acid (e.g., L-ascorbic acid), sodium metabisulfite,
ethylenediaminetetraacetic acid (EDTA) disodium, benzalkonium
chloride, sodium hydroxide, and mixtures thereof. For example, EDTA
or its salts (e.g., sodium or potassium) are employed in amounts
ranging from about 0.01% to 2% by weight of the composition
containing alkyl saccharide preservative.
[0169] Therapeutic agents or drugs of the present invention can be
peptides or proteins, medically or diagnostically useful, of small
to medium size, e.g., up to about 15 kD, 30 kD, 50 kD, 75 kD, etc.,
or a protein having between about 1-300 amino acids or more. The
methods of the invention also anticipate the use of small
molecules, for example, an organic compound that has a molecular
weight of less than 3 kD, or less than 1.5 kD.
[0170] The mechanisms of improved drug absorption according to the
invention are generally applicable and should apply to all such
peptides or protein, although the degree to which their absorption
is improved may vary according to the molecular weight (MW) and the
physico-chemical properties of the peptide or protein, and the
particular enhancer used. Examples of peptides or protein include
vasopressin, vasopressin polypeptide analogs, desmopressin,
glucagon, corticotropin (ACTH), gonadotropin, calcitonin, C-peptide
of insulin, parathyroid hormone (PTH), growth hormone (HG), human
growth hormone (hGH), growth hormone releasing hormone (GHRH),
oxytocin, corticotropin releasing hormone (CRH), somatostatin or
somatostatin polypeptide analogs, gonadotropin agonist or
gonadotrophin agonist polypeptide analogs, human atrial natriuretic
peptide (ANP), human thyroxine releasing hormone (TRH), follicle
stimulating hormone (FSH), and prolactin.
[0171] One preferred composition of the invention is the peptide
drug is Exenatide (or exendin-4) and an alkyl glycoside. Exenatide
is a synthetic version of exendin-4, and has been used in clinical
trials by Amylin Pharmaceuticals. Exendin-4 is a low molecular
weight peptide that is the first of a new class of therapeutic
medications known as incretin mimetic agents or hormones. Incretin
hormones are any of various gastrointestinal (GI) hormones and
factors that act as potent stimulators of insulin secretion, e.g.,
as gastric inhibitory polypeptide (GIP), glucagon-like peptide-1
(GLP-1), or Exenatide, or exendin-4, or equivalents thereof.
[0172] Exendin-4 is a naturally occurring 39-amino acid peptide
isolated from salivary secretions of the Gila Monster Lizard. Eng
et al., "Isolation and characterization of exendin-4, an exendin-3
analogue, from Heloderma suspectum venom. Further evidence for an
exendin receptor on dispersed acini from guinea pig pancreas," J.
Biol. Chem. 267(15):7402-7405(1992). Exenatide exhibits similar
glucose lowering actions to glucagons like peptide, or GLP-1.
Exenatide is being investigated for its potential to address
important unmet medical needs of many people with type 2 diabetes.
Clinical trials suggest that Exenatide treatment decreases blood
glucose toward target levels and is associated with weight loss.
The effects on glucose control observed with Exenatide treatment
are likely due to several actions that are similar to those of the
naturally occurring incretin hormone GLP-1 (see Example 7). These
actions include stimulating the body's ability to produce insulin
in response to elevated levels of blood glucose, inhibiting the
release of glucagon following meals and slowing the rate at which
nutrients are absorbed into the bloodstream. In animal studies
Exenatide administration resulted in preservation and formation of
new beta cells, the insulin-producing cells in the pancreas, which
fail as type 2 diabetes progresses.
[0173] Use of Exenatide, incretin mimetic agents or equivalents
thereof can be used to treat various forms of diabetes including
but not limited to brittle diabetes, chemical diabetes or impaired
glucose tolerance, gestational diabetes, diabetes insipidus,
diabetes insipidus central, diabetes insipidus nephrogenic,
diabetes insipidus pituitary, latent diabetes, lipatrophic
diabetes, maturity-onset diabetes of youth (MODY), diabetes
mellitus (DM), diabetes mellitus adult-onset (type 2 DM), diabetes
mellitus insulin-dependent (IDDM, or type 1 DM), diabetes mellitus
non-insulin dependent (NIDDM), diabetes mellitus juvenile or
juvenile-onset, diabetes mellitus ketosis-prone, diabetes mellitus
ketosis-resistant, diabetes mellitus malnutrition-related (MRDM),
diabetes mellitus tropical or tropical pancreatic, diabetes
mellitus, preclinical diabetes, or diabetes induced by various
drugs e.g., thiazide diabetes, steroid diabetes, or various
diabetes animal model including but not limited to alloxan diabetes
and puncture diabetes.
[0174] In another aspect, therapeutic compositions of the invention
are used to treat obesity. Obesity is a common problem in both
adults and adolescents. For example, PYY3-36 (or AC162352) is a
hormone that plays a critical role in decreasing appetites. The gut
hormone fragment peptide PYY3-36 (PYY) reduces appetite and food
intake when infused into subjects of normal weight. Similar to the
adipocyte hormone, leptin, PYY reduces food intake by modulating
appetite circuits in the hypothalamus. However, in obese patients
there is a resistance to the action of leptin, thereby limiting
leptin's therapeutic effectiveness. Still other studies show that
PYY reduces food intake. Injection of PYY revealed that they eat on
average 30% less than usual, resulting in weight loss. Hence, PYY
3-36 has potential as a treatment for obesity. Amylin
Pharmaceuticals submitted an Investigational New Drug application
for PYY 3-36 in 2003.
[0175] Compounds whose absorption can be increased by the method of
this invention include any compounds now known or later discovered,
in particular drugs, or therapeutic compounds, molecules or agents
that are difficult to administer by other methods, for example,
drugs that are degraded in the gastrointestinal (GI) tract or that
are not absorbed well from the GI tract, or drugs that subjects
could administer to themselves more readily via the ocular, nasal,
nasolacrimal, inhalation or pulmonary, oral cavity (sublingual or
Buccal cell), or CSF delivery route than by traditional
self-administration methods such as injection. Some specific
examples include peptides, polypeptides, proteins and other
macromolecules, for example, peptide hormones, such as insulin and
insulin analogs or derivatives such as Humalog.TM. and Novalog.TM.,
among others and calcitonin, enkephalins, glucagon and hypoglycemic
agents such as tolbutamide and glyburide, and agents which are
poorly absorbed by enteral routes, such as griseofulvin, an
antifungal agent. Other compounds include, for example, nicotine,
interferon (e.g., alpha, beta, gamma), PYY, GLP-1, synthetic
exendin-4 (Exenatide), parathyroid hormone (PTH), and human growth
hormone or other low molecular weight peptides and proteins.
[0176] As discussed herein, varying amounts of drug may be absorbed
as a drug passes through the buccal, sublingual, oropharyngeal and
oesophageal pregastric portions of the alimentary canal. However,
the bulk of the drug passes into the stomach and is absorbed in the
usual mode in which enteric dosage forms such as tablets, capsules,
or liquids are absorbed. As drug is absorbed from the intestines,
the drug is brought directly into the liver, where, depending upon
its specific chemical structure, it may be metabolized and
eliminated by enzymes that perform the normal detoxifying processes
in liver cells. This elimination is referred to as "first-pass"
metabolism or the "first-pass" effect in the liver as previously
discussed. The resulting metabolites, most often substantially or
completely inactive compared to the original drug, are often found
circulating in the blood stream and subsequently eliminated in the
urine and/or feces.
[0177] Aspects of the present invention are based on the discovery
that addition of certain alkyl saccharides, when included in
fast-dispersing dosage forms, modulate the proportion of drug that
is subject to the first-pass effect, thus allowing a fixed amount
of drug to exert greater clinical benefit, or allowing a smaller
amount of drug to achieve similar clinical benefit compared to an
otherwise larger dose.
[0178] Additional aspects of the invention are based on the
discovery that increasing or decreasing the amount of specific
alkyl saccharides included in fast-dispersing dosage forms alters
or modulates the site of absorption of a drug, increasing or
decreasing, respectively, that proportion of a drug that is
absorbed through buccal tissue compared to other portions of the
alimentary canal. In cases where it is desirable to speed the onset
of drug action but preserve the normally longer Tmax associated
with the standard oral tablet, the alkylsaccharide content can be
reduced to attenuate buccal absorption so that a portion of the
drug is immediately absorbed buccally for rapid onset, but the rest
is absorbed through the slower gastric absorption process. In this
way it has been found that by selecting an alkylsaccharide
concentration less than, for example 20% less than, the
concentration of alkylsaccharide that has been found by experiment
to produce maximal or near maximal buccal absorption, a broader
absorption peak in the "systemic drug level" vs time graph,
overall, may be achieved where this is judged to be clinically
desirable.
[0179] As further discussed in the Examples below, addition of
certain alkylsaccharides having specific alkyl chain lengths to the
fast-dispersing tablets alters the pharmacokinetics of pre-gastric
drug absorption in beneficial ways. Specifically, incorporation of
from between about 0.2%-0.3%, 0.3%-0.4%, 0.4%-0.5%, 0.5%-1.0%,
1.0%-2.0%, 2.0%-3.0%, 3.0%-4.0%, 4.0%-5.0%, 5.0%-6.0%, 6.0%-7.0%,
7.0%-8.0%, 9.0%-10.0% and greater than 10% of alkylglycoside alters
the pharmacokinetics of pre-gastric drug absorption in beneficial
ways. In exemplary embodiments, the alkylsaccharide is dodecyl
maltoside, tetradecyl maltoside and/or sucrose dodecanoate, which
when incorporated into a fast-dispersing tablet format increases
the drug that enters into systemic circulation and decreases the
drug that is eliminated by the "first-pass" effect in the liver.
Additionally, the time to maximum drug levels is dramatically
reduced, typically from one to six hours, to approximately 15 to 45
minutes. For use in treating combative patients undergoing
psychotic episodes, this more rapid absorption of drug, resulting
in more rapid onset of action, may be of great benefit.
[0180] Further, other aspects of the invention, are based on the
discovery that when certain types of fast-dissolve or
fast-dispersing tablets are placed between the cheek and gum or
into close association with buccal tissue inside the mouth, an even
larger proportion of drug is directly absorbed into systemic
circulation and a smaller amount subsequently undergoes first pass
elimination in the liver. Lastly, it has been discovered that a
particularly favorable location within the mouth for this effect is
inside the central portion of the upper lip, between the inside of
the lip and gums, directly below the nose. In exemplary aspects,
these types of fast-dissolve dosage formulations are prepared by
lyophilization or vacuum drying. In an exemplary aspect, the dosage
formulation is prepared in a manner that results in a dosage
formulation that is substantially porous.
[0181] The term "fast-dispersing dosage form" is intended to
encompass all the types of dosage forms capable of dissolving,
entirely or in part, within the mouth. However, in exemplary
aspects, the fast-dispersing dosage form is a solid,
fast-dispersing network of the active ingredient and a
water-soluble or water-dispersible carrier matrix which is inert
towards the active ingredient and excipients. In various
embodiments, the network may be obtained by lyophilizing or
subliming solvent from a composition in the solid state, which
composition comprises the active ingredient, an alkyl saccharide,
and a solution of the carrier in a solvent. While a variety of
solvents are known in the art as being suitable for this use, one
solvent particularly well suited for use with the present invention
is water. Water--alcohol mixtures may also be employed where drug
solubility in the mixed solvent is enhanced. For poorly water
soluble drugs, dispersions of small drug particles can be suspended
in an aqueous gel that maintains uniform distribution of the
substantially insoluble drug during the lyophilization or subliming
process.
[0182] In one embodiment, the aqueous gel may be the
self-assembling hydrogels described in U.S. Patent Application No.
60/957,960, formed using selected alkylsaccharides such as sucrose
mono- and di-stearate and/or tetradecyl-maltoside, incorporated
herein by reference. In various aspects, the fast-dissolve
compositions of the invention disintegrates within 20 seconds,
preferably less than 10 seconds, of being placed in the oral
cavity.
[0183] Matrix forming agents suitable for use in fast-dissolve
formulations of the present invention are describe throughout this
application. Such agents include materials derived from animal or
vegetable proteins, such as the gelatins, collagens, dextrins and
soy, wheat and psyllium seed proteins; gums such as acacia, guar,
agar, and xanthan; polysaccharides; alginates; carrageenans;
dextrans; carboxymethylcelluloses; pectins; synthetic polymers such
as polyvinylpyrrolidone; and polypeptide/protein or polysaccharide
complexes such as gelatin-acacia complexes. In exemplary aspects,
gelatin, particularly fish gelatin or porcine gelatin is used.
[0184] While it is envisioned that virtually any drug may be
incorporated into a fast-dissolve dosage formulation as described
herein, particularly well suited drugs include melatonin,
raloxifene, olanzapene and diphenhydramine.
[0185] Further, the therapeutic compositions of the invention also
contemplate non-peptide drugs or therapeutic agents. For example,
in U.S. Pat. No. 5,552,534, non-peptide compounds are disclosed
which mimic or inhibit the chemical and/or biological activity of a
variety of peptides. Such compounds can be produced by appending to
certain core species, such as the tetrahydropyranyl ring, chemical
functional groups which cause the compounds to be at least
partially cross-reactive with the peptide. As will be recognized,
compounds which mimic or inhibit peptides are to varying degrees
cross-reactivity therewith. Other techniques for preparing
peptidomimetics are disclosed in U.S. Pat. Nos. 5,550,251 and
5,288,707. The above U.S. patents are incorporated by reference in
their entirety.
[0186] The method of the invention can also include the
administration, along with the alkyl glycoside and a protein or
peptide, a protease or peptidase inhibitor, such as aprotinin,
bestatin, alpha1 proteinase inhibitor, soybean trypsin inhibitor,
recombinant secretory leucocyte protease inhibitor, captopril and
other angiotensin converting enzyme (ACE) inhibitors and thiorphan,
to aid the protein or peptide in reaching its site of activity in
the body in an active state (i.e., with degradation minimal enough
that the protein is still able to function properly). The protease
or peptidase inhibitor can be mixed with the alkyl glycoside and
drug and then administered, or it can be administered separately,
either prior to or after administration of the glycoside or
drug.
[0187] The invention also provides a method of lowering blood
glucose level in a subject comprising administering a blood
glucose-reducing amount of a composition comprising insulin and an
absorption increasing amount of a suitable nontoxic, nonionic alkyl
glycoside having a hydrophobic alkyl group joined by a linkage to a
hydrophilic saccharide, thereby increasing the absorption of
insulin and lowering the level of blood glucose. A "blood
glucose-reducing amount" of such a composition is that amount
capable of producing the effect of reducing blood glucose levels,
as taught herein. Preferred is an amount that decreases blood
glucose to normoglycemic or near normoglycemic range. Also
preferred is an amount that causes a sustained reduction in blood
glucose levels. Even more preferred is an amount sufficient to
treat diabetes, including diabetes mellitus (DM) by lowering blood
glucose level. Thus, the instant method can be used to treat
diabetes mellitus. Preferred alkyl glycosides are the same as those
described above and exemplified in the Examples.
[0188] Also provided is a method of raising blood glucose level in
a subject by administering a blood glucose-raising amount
comprising glucagons and at least one alkyl glycoside and/or
saccharide alkyl ester. When the composition includes insulin, it
can be used to cause the known effect of insulin in the
bloodstream, i.e., lower the blood glucose levels in a subject.
Such administration can be used to treat diabetes mellitus, or
related diseases. A "blood glucose-raising amount" of glucagon in
such a composition is that amount capable of producing the effect
of raising blood glucose levels. A preferred amount is that which
increases blood glucose to normoglycemic or near-normoglycemic
range. Another preferable amount is that which causes a sustained
rising of blood glucose levels. Even more preferred, is that amount
which is sufficient to treat hypoglycemia by raising blood glucose
level. Thus, this method can be used to treat hypoglycemia.
Preferred alkyl glycosides are the same as those described above
and exemplified in the Examples.
[0189] Similarly, when this composition includes glucagon, it can
be used to cause the known effect of glucagon in the bloodstream,
i.e., to raise the blood glucose levels in a subject. Such
administration can therefore be used to treat hypoglycemia,
including hypoglycemic crisis.
[0190] The invention also provides methods for ameliorating
neurological disorders which comprises administering a therapeutic
agent to the cerebral spinal fluid (CSF). The term "neurological
disorder" denotes any disorder which is present in the brain,
spinal column, and related tissues, such as the meninges, which are
responsive to an appropriate therapeutic agent. The surprising
ability of therapeutic agents of the present invention to
ameliorate the neurological disorder is due to the presentation of
the therapeutic agent to persist in the cerebro-ventricular space.
The ability of the method of the invention to allow the therapeutic
agent to persist in the region of the neurological disorder
provides a particularly effective means for treating those
disorders.
[0191] It will be understood, however, that the specific dose level
and frequency of dosage for any particular subject in need of
treatment may be varied and will depend upon a variety of factors
including the activity of the specific compound employed, the
metabolic stability and length of action of that compound, the age,
body weight, general health, sex, diet, mode and time of
administration, rate of excretion, drug combination, the severity
of the particular condition, and the host undergoing therapy.
Generally, however, dosage will approximate that which is typical
for known methods of administration of the specific compound. For
example, for intranasal administration of insulin, an approximate
dosage would be about 0.5 unit/kg regular porcine insulin (Moses et
al.). Dosage for compounds affecting blood glucose levels optimally
would be that required to achieve proper glucose levels, for
example, to a normal range of about 5-6.7 mM. Additionally, an
appropriate amount may be determined by one of ordinary skill in
the art using only routine testing given the teachings herein (see
Examples).
[0192] Furthermore, the compositions of the invention can be
administered in a format selected from the group consisting of a
drop, a spray, an aerosol and a sustained release format. The spray
and the aerosol can be achieved through use of the appropriate
dispenser. The sustained release format can be an ocular insert,
erodible microparticulates, swelling mucoadhesive particulates, pH
sensitive microparticulates, nanoparticles/latex systems,
ion-exchange resins and other polymeric gels and implants (Ocusert,
Alza Corp., California; Joshi, A., S. Ping and K. J. Himmelstein,
Patent Application WO 91/19481). These systems maintain prolonged
drug contact with the absorptive surface preventing washout and
nonproductive drug loss.
[0193] The present invention is more particularly described in the
following examples which are intended as illustrative only since
numerous modifications and variations therein will be apparent to
those skilled in the art. The following examples are intended to
illustrate but not limit the invention.
EXAMPLE 1
Alkyl Glycoside and/or Sucrose Ester Formulations do not Cause
Mucosa Irritation or Disruption
[0194] The nasal mucosa is highly vascularized and hence optimal
for high drug permeation. Moreover, absorption of drug(s) through
the nasal mucosa is available to the central nervous system (CNS).
Although local application of drugs is desirable, a challenge for
this method of administration is mucosal irritancy.
[0195] A formulation consisting of an alkyl glycoside (0.125% TDM)
in a commercial over-the-counter (OTC) nasal saline was
administered in vivo to human nasal epithelium over a period of
over one month. The 0.125% TDM formulation is compared to the
control, namely the same commercial (OTC) nasal saline, over the
same period of time. Results show that during and after 33 days of
daily TDM administration (i.e., the duration of the study), there
is no observable irritation of the nasal mucosa (data not shown).
Thus, compositions of the invention are non-toxic and non-irritable
providing repeated and long-term intranasal administration, which
is beneficial for those patients with chronic and ongoing
disease(s).
[0196] A similar test was performed using sucrose dodecanoate, a
sucrose ester.
[0197] Sucrose dodecanoate is administered in vivo to human nasal
epithelium and during and after 47 days (i.e., the duration of the
study), no observable irritation was detected (data not shown).
Thus, these results show that alkyl glycosides and sucrose esters
of the invention are non-toxic and do not cause mucosa irritation
when administered daily over a long period of time.
EXAMPLE 2
Alkyl Glycoside and/or Sucrose Ester Compositions Stabilize Drugs
by Increasing Drug Bioavailability and Reducing Drug
Bioavailability Variance
[0198] Stability of the alkyl glycoside depends, in part, on the
number of carbon atoms or length of the alkyl chain and other long
alkyl chains, with tetradecylmaltoside (TDM) having the greatest
effect; but other highly branched alkyl chains including DDM also
have stabilizing effects. In contrast to Hovgaard-1, which
described the preference for a high alkyl glycoside to drug ratio,
the instant invention shows that this ratio is much lower. For
example, alkyl glycosides in the range of about 0.01% to about 6%
by weight result in good stabilization of the drug; whereas
Hovgaard-1 shows stabilization is only achieved at much higher
ratios of alkyl glycosides to drug (10:1 and 16:1). Even more
interesting, alkyl glycosides of the invention in the range of
about 0.01% to about 6% have increased bioavailability (see FIG.
1). This is in sharp contrast to Hovgaard-2, which showed
relatively low bioavailability (0.5-1%) at the high alkyl glycoside
ratios (10:1 and 16:1).
[0199] FIG. 1 is a graph comparing the bioavailability of the drug
MIACALCIN.RTM. (salmon calcitonin from Novartis) with and without
alkyl glycoside (TDM). MIACALCIN.RTM. is a nasal spray and
administered directly onto the nasal epithelium or nasal mucosa.
FIG. 1 shows that MIACALCIN.RTM. minus alkyl glycoside has very low
bioavailability levels in humans (MIACALCIN.RTM. product
specification insert), as compared to the MIACALCIN.RTM. with alkyl
glycoside as administered to rats. More specifically, intranasal
delivery of MIACALCIN.RTM. with 0.125% and 0.250% alkyl glycoside
(TDM) resulted in about 43% to about 90% bioavailability,
respectively. The bioavailability of intranasal administration of
MIACALCIN.RTM. without alkyl glycoside is only about 3% in humans,
and was undetectable in rats, suggesting that the rat is a
stringent model for estimating intranasal drug absorption in
humans. Thus, the alkyl glycoside of the invention enhances
absorption and increases bioavailability of the drug.
[0200] Furthermore, besides increasing the bioavailability of the
drug, the alkyl glycoside compositions of the invention effectively
decrease the bioavailability variance of the drug. FIG. 1 shows
that administration of MIACALCIN.RTM. with alkyl glycoside (0.125%
or 0.25%) intranasally has a bioavailability variance of +/-8%,
whereas the bioavailability variance without alkyl glycoside is
0.3% to 30%, or a two orders of magnitude change. The increase in
bioavailability and the decrease in the bioavailability variance
ensures patient-to-patient variability is also reduced. The results
as shown in FIG. 1 are administered intranasally, however, similar
results are expected for oral, buccal, vaginal, rectal, etc.
delivery and at different alkyl glycoside concentrations.
[0201] Thus, contrary to the art, the alkyl glycoside compositions
of the invention, in the range of about 0.01% to about 6% result in
increased bioavailability and reduced bioavailability variance.
This has not otherwise been reported.
EXAMPLE 3
Ocular Administration of Alkyl Saccharides Plus Insulin Produces
Hypoglycemic Effects In Vivo
[0202] Normal rats were anesthetized with a mixture of
xylazine/ketamine to elevate their blood glucose levels. The
elevated levels of D-glucose that occur in response to anesthesia
provide an optimal system to measure the systemic hypoglycemic
action of drug administration, e.g., insulin-containing eye drops.
This animal model mimics the hyperglycemic state seen in diabetic
animals and humans. In the experimental animal group, anesthetized
rats are given eye drops containing insulin. Blood glucose levels
from the experimental group are compared to anesthetized animals
which received eye drops without insulin. The change in blood
glucose levels and the differential systemic responses reflects the
effect of insulin absorbed via the route of administration, e.g.,
ocular route.
[0203] Adult male Sprague-Dawley rats (250-350 g) were fed ad
libitum, and experiments were conducted between 10:00 a.m. and 3:00
p.m. Rats were anesthetized with a mixture of xylazine (7.5 mg/kg)
and ketamine (50 mg/kg) given intraperitoneally (IP) and allowed to
stabilize for 50-90 min before the administration of eye drops.
Anesthesia of a normal rat with xylazine/ketamine produces an
elevation in blood glucose values which provides an optimal state
to determine the systemic hypoglycemic action of insulin-containing
eye drops. Blood D-glucose values were measured by collecting a
drop of blood from the tail vein at 5-10 min intervals throughout
the experiment and applying the blood to glucometer strips
(Chemstrip bG) according to directions provided with the instrument
(Accu-Chek II, Boehringer Mannheim Diagnostics; Indianapolis,
Ind.). Blood D-glucose values ranged from 200 to 400 mg/dl in
anesthetized nondiabetic rats.
[0204] At time 0, after a 50-90 min stabilization period, rats were
given 20 .mu.l of eye drops composed of phosphate-buffered saline
(PBS) with or without 0.2% regular porcine insulin and 0.125%-0.5%
of the absorption enhancing alkyl glycoside (e.g., TDM) to be
tested. Eye drops were instilled at time 0 using a plastic
disposable pipette tip with the eyes held open, and the rat was
kept in a horizontal position on a warming pad (37.degree. C.)
throughout the protocol. The rats were given additional anesthesia
if they showed signs of awakening. Rats received in each eye 20
.mu.l of 0.125-0.5% absorption enhancer in phosphate buffered
saline, pH 7.4 with (experimental) or without (control) 0.2% (50
U/ml) regular porcine insulin (Squibb-Novo, Inc.) for a total of 2
U per animal. Octyl-.beta.-D-maltoside, decyl-.beta.-D-maltoside,
dodecyl-.mu.-D-maltoside, tridecyl-.beta.-D-maltoside and
tetradecyl-.beta.-D-maltoside were obtained from Anatrace, Inc.
(Maumee, Ohio). Hexylglucopyranoside, heptylglucopyranoside,
nonylglucopyranoside, decylsucrose and dodecylsucrose were obtained
from Calbiochem, Inc. (San Diego, Calif.); Saponin, BL-9 and Brij
78 were obtained from Sigma Chemical Co. (St. Louis, Mo.).
[0205] The D-glucose levels in the blood remained elevated when the
animals received eye drops containing: 1) saline only; 2) 0.2%
regular porcine insulin in saline only; or 3) absorption enhancer
only. However, when rats received eye drops containing 0.2% regular
porcine insulin and several alkylmaltoside or alkylsucrose
compounds, a pronounced decrease in blood D-glucose values occurred
and was maintained for up to two hours. Insulin administered
ocularly with 0.5% dodecyl-.beta.-D-maltoside (see Table I) or 0.5%
decyl-.beta.-D-maltoside (see Table III) results in a prompt and
sustained fall in blood glucose levels which are maintained in the
normoglycemic (80-120 mg/dl) or near-normoglycemic (120-160 mg/dl)
range for the two hour duration of the experiment. Hence, at least
two alkylmaltosides are effective in achieving sufficient
absorption of insulin delivered via the ocular route to produce a
prompt and sustained fall in blood glucose levels in experimentally
hyperglycemic animals. The surfactant compositions of the invention
are therefore useful to achieve systemic absorption of insulin and
other peptides/proteins, e.g., glucagon and macromolecular drugs
and heparin delivered via the ocular route in the form of eye
drops.
[0206] Several other alkylmaltosides are also effective as
absorption enhancers for ocular administration of insulin including
0.5% tridecylmaltoside (see Table III) and 0.125% (Table II) and
0.5% tetradecyl maltoside. These studies show that alkylmaltosides
with the longer alkyl chains (or number of carbon atoms), e.g.,
dodecyl-, tridecyl- and tetradecyl-.beta.-D-maltosides, are more
effective. The increase in the number of carbon atoms also
contributes to the greater hydrophobic/hydrophilic structural
balance and absorption enhancing effect. The shorter alkyl chains
(fewer carbon atoms) e.g., decylmaltoside, or no, e.g.,
octylmaltoside, produce less absorption enhancing activity. It is
noted that the most effective alkylmaltosides produce effects
comparable to or greater than those seen with other absorption
enhancers such as saponin, and with the added advantage that they
can be metabolized to nontoxic products following systemic
absorption.
[0207] The effects of the alkylmaltosides as absorption enhancers
are dose-dependent, as can be seen by examining the effects of
different concentrations ranging from 0.125-0.5% in producing a
hypoglycemic effect when combined with insulin. Whereas, 0.5% and
0.375% dodecylmaltoside appear equally effective in achieving
systemic absorption of insulin and reduction of blood glucose
levels, 0.25% has a smaller and more transient effect and 0.125% is
ineffective (Table I). Similarly, tridecylmaltoside also shows a
dose-dependent effect in lowering blood glucose concentrations when
combined with insulin, but the effect achieved with even 0.25% of
the absorption enhance is sustained for the two hour time course of
the experiment. Thus, dose-dependent effects of the alkylmaltosides
suggest that they achieve enhancement of protein absorption via the
ocular route in a graded fashion proportional to the concentration
of the agent.
TABLE-US-00001 TABLE I Effect of Eye Drops Containing Insulin Plus
Various Concentrations of Dodecyl Maltoside on Blood Glucose Values
(in mg/dl) in Rat Dodecyl Maltoside Concentration 0.125% 0.25%
0.375% 0.50% Time (min) Blood Glucose Concentrations (mg/dl) -20
305 .+-. 60 271 .+-. 38 305 .+-. 51 375 .+-. 9 -10 333 .+-. 58 295
.+-. 32 308 .+-. 27 366 .+-. 12 0 338 .+-. 67 323 .+-. 62 309 .+-.
32 379 .+-. 4 30 349 .+-. 64 250 .+-. 48 212 .+-. 18 297 .+-. 18 60
318 .+-. 38 168 .+-. 22 134 .+-. 4 188 .+-. 25 90 325 .+-. 57 188
.+-. 55 125 .+-. 12 141 .+-. 13 120 342 .+-. 78 206 .+-. 63 119
.+-. 19 123 .+-. 5
[0208] The absorption enhancing effects of the alkyl saccharides
were not confined to the alkylmaltosides alone since dodecylsucrose
(0.125%, 0.25%, 0.375%) also shows a dose-dependent effect in
producing ocular absorption of insulin and reduction in blood
glucose levels. This effect is observed even at 0.125% alkyl
saccharide (from 335 mg/dl.+-0.26 mg/dl at time 0 min. to 150
mg/dl+-0.44 mg/dl at time 120 min.). 0.5% decylsucrose was also
effective in reducing blood glucose levels, but as shown for the
alkylmaltosides, a reduction in the length of the alkyl chain, and
hence the hydrophobic properties of the molecule, appears to reduce
the potency of the alkylsucrose compounds. However, a significant
and sustained reduction in blood glucose levels is achieved with
0.5% decylsucrose (from 313 mg/dl.+-0.15 mg/dl at time 0 min. to
164 mg/dl+-0.51 mg/dl at time 120 min.). The absorption enhancing
abilities of alkyl saccharides with two distinct disaccharide
moieties suggests that it is the physicochemical properties of the
compounds which are crucial to their activity and that other alkyl
saccharides, e.g., dodecyllactose, have the right balance of
properties to be equally or more effective as absorption enhancers
while retaining the metabolic and nontoxic properties of the
alkylsaccharide enhancing agents. These alkyl saccharides are
anticipated by the invention.
[0209] Studies with alkylglucosides were also conducted; 0.5%
hexylglucoside and 0.5% heptylglucoside were ineffective at
promoting insulin absorption from the eye, but 0.5% nonylglucoside
effectively stimulated insulin absorption and reduced blood glucose
levels (from 297 mg/dl to 150 mg/dl). This result once further
supports that the alkyl chain length, as well as the carbohydrate
moiety, play critical roles in effectively enhancing insulin
absorption.
[0210] It should be noted that no damaging effects (i.e.,
non-irritants) to the ocular surface were observed with any of the
alkylmaltoside or alkylsucrose agents employed in these studies.
Furthermore, the prompt and sustained hypoglycemic effects produced
by these agents in combination with insulin suggest that these
absorption enhancers do not adversely affect the biological
activity of the hormone, in keeping with their nondenaturing, mild
surfactant properties.
[0211] Thus, therapeutic compositions on the invention consisting
of at least an alkyl glycoside and a drug are stable and the alkyl
glycosides enhance the absorption of the drug.
EXAMPLE 4
Ocular and Intranasal Administration of TDM Plus Glucagon Produces
Hypoglycemic Effects In Vivo
[0212] Since previous Examples showed that administration via eye
drops of an absorption enhancer with drug e.g., insulin results in
significant absorption of the drug via the nasolacrimal drainage
system, therapeutically effective administration of insulin with
alkylmaltosides, alkylsucrose and like agents by intranasal
administration is tested herein.
[0213] Tetradecylmaltoside (TDM) in combination with insulin also
produced a drop in blood D-glucose levels when administered in the
form of a drop intranasally as well as via a drop by the ocular
route. Eye drops containing 0.2% regular porcine insulin with
0.125% tetradecylmaltoside are administered to rats as previously
described. The administration of the composition produces a prompt
and prominent drop in blood glucose levels. The drop in blood
glucose levels decrease even more by administration of a nose drop
containing the same concentration of insulin with 0.5%
tetradecylmaltoside (Table II). Thus, intranasal delivery and
administration of the alkyl saccharide with drug results in
lowering of blood glucose levels.
TABLE-US-00002 TABLE II Effect of Insulin Eye Drops, Containing
0.125% Tetradecyl Maltoside and Nose Drops Containing 0.5%
Tetradecyl Maltoside on Blood Glucose Values in Rats Blood Glucose
Time (min) (mg/dl) -20 319 -10 311 Eye drops added 0 322 15 335 30
276 45 221 60 212 75 167 90 174 105 167 120 208 Nose Drops Added
135 129 150 74 165 76 180 68
EXAMPLE 5
Ocular Administration of Alkyl Saccharides Plus Insulin Produces
Hyperglycemic Effects In Vivo
[0214] Previous studies demonstrated that insulin absorption from
the eye is stimulated by saponin, BL-9 and Brij-78. BL-9 and
Brij-78 are ineffective at stimulating the absorption of glucagon
from the eye, whereas saponin is effective. Glucagon absorption
from the eye was measured in rats given eye drops containing
various surfactants plus glucagon (30 rig) (Eli Lilly,
Indianapolis, Ind.) by monitoring an elevation in blood D-glucose
levels. In these experiments, rats were anesthetized with sodium
pentobarbital rather than xylazine/ketamin. This modification of
the procedure resulted in basal blood glucose levels in the
normoglycemic range and made it possible to readily monitor the
hyperglycemic action of any glucagon absorbed from the eye.
[0215] Paired animals that receive eye drops containing the
surfactant alone, or glucagon alone, were compared to animals
receiving eye drops with the surfactant plus glucagon. When
eyedrops containing 0.5% saponin plus glucagon are administered to
rats, the level of D-glucose in blood rises significantly, but no
such effect is observed with eye drops containing 0.5% BL-9 or 0.5%
Brij-78 plus glucagon. Interestingly, when eye drops containing
dodecylsucrose, decylmaltose or tridecylmaltose plus glucagon are
administered to rats which were previously treated with eye drops
containing these surfactant agents plus insulin, the glucagon is
absorbed and blood D-glucose values increase significantly (Table
III). This result confirms that ocular administration of certain
alkylsaccharides can enhance the absorption of drugs, including
glucagon and insulin. Moreover, it is now possible to treat for a
hypoglycemic crisis using a formulation with at least an alkyl
saccharide of the invention.
TABLE-US-00003 TABLE III Effect of Eye Drops Containing Insulin or
Glucagon and 0.5% Decyl Maltoside, 0.5% Dodecyl Sucrose, or 0.5%
Tridecyl Maltoside on Blood Glucose Values in Rats Surfactant Agent
Dodecyl Decyl Tridecyl Time Sucrose Maltoside Maltoside (min) Blood
Glucose Concentration (mg/dl) -20 266 249 255 -10 305 287 307
Insulin Eye Drops Added 0 351 337 323 10 347 304 309 20 252 292 217
30 161 221 131 40 120 164 100 50 105 138 87 60 114 114 107 70 113
104 115 80 104 110 79 90 86 120 85 100 113 92 76 110 107 81 74 120
112 87 75 Glucagon Eye Drops Added 130 111 95 82 140 143 99 121 150
202 132 148 160 247 157 173 170 242 171 162 180 234 180 162 190 211
189 156
EXAMPLE 6
Intranasal Administration of 0.25% TDM Plus Insulin Decreases Blood
Glucose Levels In Vivo
[0216] Intranasal administration of drugs or agents are possible in
animal models e.g., mice and rats, although the nasal opening in is
very small. In the experiments and results described herein, an
anesthesia-induced hyperglycemia model was used (described in
Examples above). Hyperglycemic animals were induced by an
intraperitoneal (IP) injection containing xylazine-ketamine and
blood glucose levels were monitored over a period of time.
Immediately after the xylazine-ketamine injection, there was an
increase in the blood glucose levels as shown in FIG. 2 (closed
dark circles), and blood glucose levels were about 450 mg/dl. The
increase in blood glucose levels was attributed to the inhibition
of pancreatic insulin secretion. Blood glucose levels peak to about
482 mg/dl by 30 minutes after the xylazine-ketamine injection (FIG.
2). Then, at approximately 33 minutes after the xylazine-ketamine
injection, 6 .mu.L of insulin (Humalog) in 0.25%
tetradecylmaltoside (TDM; or Intravail A) was administered
intranasally using a long thin micropipette tip, and blood glucose
levels were monitored at about 15 minute intervals. After
administration of the 0.25% TDM/insulin composition, there was a
rapid decrease in blood glucose levels, reaching a low of about 80
mg/dl at about the 60 minute time point, or about 30 minutes after
the insulin administration (FIG. 2). At about the 75 minute time
point, blood glucose levels gradually returned to the baseline
level in a normoglycemic mouse, or about 80-100 mg/dl.
[0217] The results above were compared with animals treated with
insulin alone (same dosage), minus 0.25% TDM (FIG. 2, open
circles). The insulin only treatment showed blood glucose levels do
not start to decline until at about the 120 minute time mark, or
about 110 minutes after the insulin administration. Further, the
blood glucose levels observed in animals treated with insulin alone
never return to normoglycemic levels, as was observed in those
animals receiving insulin plus 0.25% TDM (FIG. 2).
[0218] Thus, these results again demonstrate that compositions of
the invention consisting of certain alkyl glycosides or alkyl
saccharides plus a drug, e.g., insulin, effectively lower blood
glucose levels, and that these effects are measurable shortly after
administration of the drug.
EXAMPLE 7
Intranasal Administration of 0.25% TDM (Intravail A)+Exendin-4
Decreases Blood Glucose Levels In Vivo
[0219] The ob/ob mouse model was utilized for the studies described
herein. Friedman, J. M., Nature 404, 632-634 (2000). All animals
received an intraperitoneal (IP) injection of a bolus of 2 g/kg
glucose for purposes of determining glucose tolerance. At time 0
the experimental animals were given about 100 micrograms/kg of
exendin-4/0.25% TDM (exendin-4 from American Peptide) either as 10
.mu.l of nasal drops (FIG. 3; closed triangles), or by IP injection
(FIG. 3; closed circles), or by and IP injection of saline alone
(no drug, no TDM; FIG. 3; open circles). Control animals were
previously performed and received no drugs. The results of this
study are shown in FIG. 3.
[0220] FIG. 3 shows that glucose tolerance of the animals were
different since blood glucose levels vary at time 0 when the
animals received the glucose bolus. Regardless, of the glucose
tolerance level at time 0, immediately after injection of the
glucose bolus, blood glucose levels increased in all three animals.
The blood glucose level of the animal receiving the IP injection of
saline alone does not decrease as rapidly as the experimental
animals receiving the drug. Moreover, the animal receiving the IP
injection of saline alone never reached a normoglycemic level (FIG.
3, open circles). In contrast, the experimental animals, after
administration of nasal drops of exendin-4/TDM, or IP injection of
exendin-4/TDM, showed a rapid and immediate decrease in blood
glucose levels.
[0221] Also exendin-4 administered about 15-30 minutes ahead of the
glucose bolus (before time 0 in FIG. 3; data not shown) produced an
even more pronounced lowering of blood glucose effect, because the
absorption of the hormone takes a certain amount of time to be
absorbed and to be active. Thus, exendin-4 (or Exenatide) which is
currently in human clinical trials, when combined with alkyl
glycosides of the invention, effectively treats a hyperglycemic
condition by lowering the blood glucose levels of the hyperglycemic
subject.
EXAMPLE 8
Alkylglycosides have Antibacterial Activity by Reducing Bacterial
Log Growth
[0222] The cultures of Candida albicans (ATCC No. 10231),
Aspergillus niger (ATCC No. 16404), Escherichia coli (ATCC No.
8739), Pseudomonas aeruginosa (ATCC No. 9027), and Staphylococcus
aureus (ATCC No. 6538) were obtained from American Type Culture
Collection, 10801 University Boulevard, Manassas, Va. 20110-2209.
The viable microorganisms used in the invention were not more than
five passages removed from the original ATCC culture. As described
herein, one passage is defined as the transfer of organisms from an
established culture to fresh medium and all transfers are
counted.
[0223] Cultures received from the ATCC are resuscitated according
to the directions provided by the ATTC. Cells grown in broth were
pelleted by centrifugation, resuspended in 1/20th the volume of
fresh maintenance broth, and combined with an equal volume of 20%
(v/v in water) sterile glycerol. Cells grown on agar were scraped
from the surface into the maintenance broth also containing 10%
glycerol broth. Small aliquots of the suspension were dispensed
into sterile vials and the vials were stored in liquid nitrogen or
in a mechanical freezer at a temperature no higher than about
-50.degree. C. When a fresh seed-stock vial was required, it was
removed and used to inoculate a series of working stock cultures.
These working stock cultures were then used periodically (each day
in the case of bacteria and yeast) to start the inoculum
culture.
[0224] All media described herein should be tested for growth
promotion using the microorganisms indicated above under Test
Organisms.
[0225] To determine whether the alkyl saccharides of the invention
inhibit growth or have antibacterial activity, the surface of a
suitable volume of solid agar medium was inoculated from a fresh
revived stock culture of each of the specified microorganisms. The
culture conditions for the inoculum culture is substantially as
described in Table IV. For example, suitable media can include but
is not limited to, Soybean-Casein Digest or Sabouraud Dextrose Agar
Medium. The bacterial and C. albicans cultures was harvested using
sterile saline TS, by washing the surface growth, collecting it in
a suitable vessel, and adding sufficient sterile saline TS to
obtain a microbial count of about 1.times.10.sup.8 colony-forming
units (cfu) per mL. To harvest the cells of A. niger, a sterile
saline TS containing 0.05% of polysorbate 80 was used, and then
adding sufficient sterile saline TS to obtain a count of about
1.times.108 cfu per mL.
[0226] Alternatively, the stock culture organisms may be grown in
any suitable liquid medium (e.g., Soybean-Casein Digest Broth or
Sabouraud Dextrose Broth) and the cells harvested by
centrifugation, and washed and resuspended in sterile saline TS to
obtain a microbial count of about 1.times.108 cfu per mL. The
estimate of inoculum concentration was determined by turbidimetric
measurements for the challenge microorganisms. The suspension
should be refrigerated if it is not used within 2 hours. To confirm
the initial cfu per mL estimate, the number of cfu per mL in each
suspension was determined using the conditions of media and
microbial recovery incubation times listed in Table IV (e.g., from
about 3 to about 7 days). This value serves to calibrate the size
of inoculum used in the test. The bacterial and yeast suspensions
were used within 24 hours of harvest; whereas the fungal
preparation can be stored under refrigeration for up to 7 days.
TABLE-US-00004 TABLE IV Culture Conditions for Inoculum Preparation
Microbial Inoculum Recovery Incubation Incubation Incubation
Organism Suitable Medium Temperature Time Time Escherichia coli
Soybean-Casein Digest Broth; 32.5 .+-. 2.5 18 to 24 hours 3 to 5
days (ATCC No. 8739) Soybean-Casein Digest Agar Staphylococcus
aureus Soybean-Casein Digest Broth; 32.5 .+-. 2.5 18 to 24 hours 3
to 5 days (ATCC No .6538) Soybean-Casein Digest Agar Candida
albicans Sabouraud Dextrose Agar; 22.5 .+-. 2.5 44 to 52 hours 3 to
5 days (ATCC No. 10231) Sabouraud Dextrose Broth Aspergillus niger
Sabouraud Dextrose Agar; 22.5 .+-. 2.5 6 to 10 days 3 to 7 days
(ATCC No. 16404) Sabouraud Dextrose Broth
[0227] To determine which alkylglycoside formulations have
antibacterial activity, the formulations were prepared in phosphate
buffered saline (PBS) at pH 7. As a source of nutrition, either 1.5
mg/mL bovine serum albumin (BSA; see Tables V and VI) or 1 mg/mL of
PYY was added (see Table VII) to the medium. BSA (CAS Number:
9048-46-8) was obtained from Sigma-Aldrich, St. Louis, Mo., USA,
n-dodecyl-4-O-.alpha.-D-glucopyranosyl-.beta.-D-glucopyranoside and
n-tetradecyl-4-O-.alpha.-D-glucopyranosyl-.beta.-D-glucopyranoside
were obtained from Anatrace Inc., Maumee, Ohio, USA, and PYY was
obtained from Bachem California Inc., Torrance, Calif., USA.
[0228] Antibacterial activity of the alkylglycosides were conducted
in four sterile, capped bacteriological containers of suitable size
into which a sufficient volume of alkylglycoside solution had been
transferred. Each container was inoculated with one of the prepared
and standardized inoculums and mixed. The volume of the suspension
inoculum was between about 0.5% and about 1.0% of the volume of the
alkylglycoside solution. The concentrations of test microorganisms
added to the alkylglycoside solution was such that the final
concentrations of the test preparation after inoculation was
between about 1.times.10.sup.5 and 1.times.10.sup.6 cfu per mL of
alkylglycoside solution. To determine the level of inhibition of
growth, or reduction of growth based on a logarithmic scale, the
initial concentration of viable microorganisms in each test
preparation was estimated based on the concentration of
microorganisms in each of the standardized inoculum as determined
by the plate-count method. The inoculated containers were then
incubated at about 22.5.degree. C..+-.2.5. The growth or non-growth
of the microorganisms in each culture/container were again
determined at day 14 and day 28. The number of cfu present in each
calculation was determined by the plate-count procedure standard in
the art for the applicable intervals. The change in the orders of
magnitude of bacterium and/or fungi was then determined by
subtracting the first calculated log 10 values of the
concentrations of cfu per mL present at the start or beginning
(e.g., day 0), from the log 10 values of the concentration of cfu
per mL for each microorganism at the applicable test intervals
(e.g., day 14 and day 28; see Tables V, VI and VII).
TABLE-US-00005 TABLE V Log Reduction of Microorganisms in Cultures
Containing 0.125%
n-Dodecyl-4-O-.alpha.-D-glucopyranosyl-.beta.-D-glucopyranoside
Staphlococcus Escherichia Candida Aspergillus aureus Coli albicans
niger Day 0 7.3 x 10.sup.5 1.2 .times. 10.sup.5 3.2 .times.
10.sup.5 4.8 .times. 10.sup.5 (cfu/gm) (cfu/gm) (cfu/gm) (cfu/gm)
Day 14 .gtoreq.5.2 orders of N.D. 3.0 orders of 0.7 orders of
magnitude magnitude magnitude reduction reduction reduction Day 28
.gtoreq.5.2 orders of 0.1 orders of .gtoreq.5.3 orders of No growth
from magnitude magnitude magnitude initial count reduction
reduction reduction
TABLE-US-00006 TABLE VI Log Reductions in Cultures Containing 0.2%
n-Tetradecyl-4-O-.alpha.-D-glucopyranosyl-.beta.-D-glucopyranoside
Staphlococcus Escherichia Candida Aspergillus aureus Coli albicans
niger Day 0 7.3 .times. 10.sup.5 1.2 .times. 10.sup.5 3.2 .times.
10.sup.5 4.8 .times. 10.sup.5 (cfu/gm) (cfu/gm) (cfu/gm) (cfu/gm)
Day 14 .gtoreq.5 orders of N.D. 3.0 orders of 0.5 orders of
magnitude magnitude magnitude reduction reduction reduction Day 28
.gtoreq.5 orders of No growth .gtoreq.5.4 orders of No growth from
magnitude from initial magnitude initial count reduction count
reduction
TABLE-US-00007 TABLE VII Log Reduction of Cultures Containing 0.25%
n-Dodecyl-4-O-.alpha.-D-glucopyranosyl-.beta.-D-glucopyranoside
Staphlococcus Escherichia Candida Aspergillus aureus Coli albicans
niger Day 0 7.3 .times. 10.sup.5 1.2 .times. 10.sup.5 3.2 .times.
10.sup.5 4.8 .times. 10.sup.5 (cfu/gm) (cfu/gm) (cfu/gm) (cfu/gm)
Day 14 .gtoreq.4.9 orders of .gtoreq.5 orders of .gtoreq.4.5 orders
of 4.7 orders of magnitude magnitude magnitude magnitude reduction
reduction. reduction. reduction Day 28 .gtoreq.4.9 orders of
.gtoreq.5 orders of .gtoreq.4.5 orders of 4.7 orders of magnitude
magnitude magnitude magnitude reduction reduction. reduction.
reduction
[0229] Determining the antibacterial activity of other
alkylglycosides would occur substantially as described herein.
EXAMPLE 9
Administration of Alkylglycosides with Antisense Oligonucleotides
to Primates
[0230] An approximately 7,000 Dalton antisense oligonucleotide
(ASO) with a modified backbone (phosphorothioate oligonucleotide as
described in U.S. Pat. No. 7,132,530) mixed with alkylglycoside
tetradecyl-beta-D-maltoside (Intravail.TM.), was administered to
six Cynomolgus monkeys cannulated into the jejunum at a dose of 10
mg/kg. The animals were fasted prior to administration. Test agents
were dissolved in PBS buffer and injected through the cannula into
the jejunum of each animal in a 1.5 mL volume or administered
subcutaneously (s.c.) as noted in Table VIII.
TABLE-US-00008 TABLE VIII Bioavailability of Antisense Drugs
Administered With Tetradecyl-Beta-D-Maltoside Average
Bioavailability Test Agents (n = 6) Observations ASO (no
tetradecyl- 0% Intestinal pili completely intact beta-D-maltoside)
(undetectable) intrajejunal ASO administered s.c. 100% Intestinal
pili completely intact at 0.5 mg/kg. 10 mg ASO + 50 18% +/- 7%
Intestinal pili completely intact mg/kg tetradecyl-
beta-D-maltoside A5 intrajejunal 10 mg ASO + 9% +/- 7% The tops of
some of the intestinal 50 mg/kg sodium pili were found to be
missing caprate intrajejunal
[0231] The protocol involved a 3 way crossover in which each animal
had the first 3 test agents in Table VIII administered on 3
different dates. There was a 1 week washout period between dosing
dates. Two of the animals were subsequently given a fourth test
agent containing 5% sodium caprate as an absorption enhancer.
Analysis of the blood levels was conducted using quantitative
analysis involving solid phase extraction using cationic
polystyrene nanoparticles.
[0232] Solid-phase extractions of the blood samples were first
performed. Nanoparticle-oligonucleotide conjugates were formed
using a known amount of oligonucleotide added to an aliquot of each
sample (200-400 .mu.l) and diluted with 800 .mu.l of 50 mM Tris HCl
(pH 9) in deionized water. The mixture was briefly vortexted prior
to the addition of 200 .mu.l of a polystyrene nanoparticle
suspension prepared by surfactant-free emulsion polymerization
using water-soluble cationic initiators to induce a positive
surface charge (solid content: approximately 10 mg/ml). The mixture
was subsequently vortexed again. After 5-10 min of incubation, the
suspension was centrifuged and the supernatant removed. The
particles were resuspended in 1 ml of a solution of 0.5 M acetic
acid in deionized water/ethanol (1:1) and separated from the
washing solution by centrifugation. After the supernatant was
removed, the particles were resuspended in 1 ml of deionized water
and separated by another centrifugation step. 200 .mu.l of a
solution of 150 .mu.M SDS in aqueous ammonia (25%)/acetonitrile
(60/40) was added to the nanoparticle-oligonucleotide conjugates
and the released oligonucleotides were separated from the carrier
by centrifugation. In order to exclude contamination of the samples
with residual particles, the supernatant was placed in another
1.5-ml tube and centrifuged again. Subsequently, the samples were
dried by rotoevaporation or lyophilization and stored at
-20.degree. C. until analysis.
[0233] Quantitative analysis was performed with capillary gel
electrophoresis of the extracted samples. Capillary gel
electrophoresis (CGE) was performed with a capillary
electrophoresis system. An oligonucleotide analysis kit containing
polyvinyl alcohol (PVA) coated capillaries, polymer solution B, and
oligonucleotide buffer was obtained. Using PVA-coated capillaries,
analysis was carried out using the manufactures protocol.
[0234] Using the data obtained from CGE analysis, quantitation of
phosphorothioate oligonucleotides was carried out. The amount of
oligonucleotides in the samples (nON) was calculated using the
following formula:
n.sub.ON=n.sub.Std(.epsilon..sub.Std/.epsilon..sub.ON)((A.sub.ON/T.sub.O-
N)/(A.sub.Std/T.sub.Std)),
where n.sub.Std is the amount of standard oligonucleotide added to
the sample, .epsilon..sub.Std and .epsilon..sub.ON are the molar
extinction coefficients, and A.sub.Std/T.sub.Std and
A.sub.ON/T.sub.ON are the corrected peak areas (quotient of peak
area and migration time) of the standard and the investigated
compound, respectively. The quotient of the corrected peak areas of
the analyte and the standard is referred to as the normalized
area.
[0235] AUC's were calculated from the concentration vs. time cures
over a 240 minute period. The relative bioavailabilities were
determined as the ratio of each AUC divided by the AUC for the
intravenously administered drug. Intravail.TM.
(tetradecyl-beta-D-maltoside) excipient provided bioavailability up
to 18%. The control showed no detectable absorption without a
surfactant excipient. The sodium caprate formulation showed an
average bioavailability of 9%.
EXAMPLE 10
Preparation of Fast-Dispersing Dosage Forms of Olanzapine
[0236] Fast-dispersing dosage forms of olanzapine were prepared as
follows. Olanzapine, CAS #132539-06-1, is obtained from SynFine
(Ontario, Canada). Sodium acetate buffer, 10 mM, pH 5.0 and pH 6.5
is prepared as follows. In an appropriate sized clean container
with volumetric markings, place 495 mL of sterile water for
injection. Add 0.286 mL acetic acid. Add 1N NaOH to bring the pH to
5.00 (or to pH 6.5). When the proper pH is obtained, add additional
water to bring the total volume to 500 mL and recheck the pH.
[0237] Liquid formulations having the compositions illustrated in
Table IX below are made up by adding the fish gelatin or porcine
skin gelatin slowly to the acetate buffer and allowing sufficient
time to dissolve while stirring throughout the process. Upon
complete dissolution of the fish or porcine skin gelatin, the
mannitol is added and allowed to dissolve. Then the sweetener is
added. Once this has been fully dispersed, the active ingredient,
olanzapine, being one of the examples for the compounds of the
present invention, is added to produce the final solution.
Secondary components such as preservatives, antioxidants,
surfactants, viscosity enhancers, coloring agents, flavoring
agents, sweeteners or taste-masking agents may also be incorporated
into the composition. Suitable coloring agents may include red,
black and yellow iron oxides and FD & C dyes such as FD & C
blue No. 2 and FD & C red No. 40 available from Ellis &
Everard. Suitable flavoring agents may include mint, raspberry,
licorice, orange, lemon, grapefruit, caramel, vanilla, cherry and
grape flavors and combinations of these. Suitable sweeteners
include aspartame, acesulfame K and thaumatin. Suitable
taste-masking agents include sodium bicarbonate. Cyclodextrins
should be avoided since they form inclusion compounds with
alkylsaccharides that reduce the effectiveness of these
excipients.
[0238] Aliquots of 1 mL each of the above drug solutions are placed
in the wells of a 24 well disposable microwell plastic plate. The
micro well plate containing the liquid aliquots is frozen at
-70.degree. in the frozen plate is placed within a glass
lyophilization flask attached to a LabConco Freezone Model 4.5
desktop freeze drier and lyophilized under vacuum. Following
lyophilization, the rapidly dispersing tablets are stored in the
micro well plate in a dry environment until tested. Sucrose mixed
mono- and di-stearate was provided as a gift by Croda Inc. and is
designated CRODESTA F-110. Dodecyl maltoside, tetradecyl maltoside
and sucrose mono-dodecanoate is obtained from Anatrace Inc.,
Maumee, Ohio.
TABLE-US-00009 TABLE IX Olanzopine Formulations Example No.
Ingredients 1 2 3 4 5 6 7 Olanzapine.sup.1 62.5 mg 125 mg 250 mg
500 mg 125 mg 250 mg 500 mg Fish Gelatin .sup.2 1.3 g 1.3 g 1.3 g
1.3 g -- -- -- (3101) Dodecyl 250 mg 500 mg -- -- 250 mg 500 mg --
maltoside.sup.3 Sucrose -- -- 250 mg 750 mg -- -- 750 mg
dodecanoate.sup.4 Gelatin Type A.sup.5 -- -- -- -- 1.3 g 1.5 2.0
Mannitol 1 g 1 g 1 g 1 g 1 g 1 g 1 g EP/USP Acesulfame K 0.062 g
0.062 g 0.062 g 0.062 g -- -- -- Aspartame -- -- -- -- 0.125 g
0.125 g 0.125 g Acetate Buffer Q.sub.s Q.sub.s Q.sub.s Q.sub.s
Q.sub.s Q.sub.s Q.sub.s (mL) 25 mL 25 mL 25 mL 25 mL 25 mL 25 mL 25
mL .sup.1SynFine, Ontario, Canada .sup.2 Croda Colloids Ltd
(non-hydrolysed, spray dried fish gelatin) .sup.4Sucrose
dodecanoate (monoester) - Anatrace Inc. .sup.5Sigma Aldrich
(Gelatin Type A, porcine skin -G6144) Q.sub.s = sufficient to
give.
[0239] The drug olanzapine, also called Zyprexa, is known to be
well absorbed when administered as a "whole-swallowed" tablet and
reaches peak concentrations in approximately 6 hours following an
oral dose. It is eliminated extensively by first pass metabolism,
with approximately 40% of the dose metabolized before reaching the
systemic circulation. Pharmacokinetic studies showed that
"whole-swallowed" olanzapine tablets and rapidly dispersing
olanzapine tablets prepared by lyophilization in the manner
described above in this Example, which disintegrate in about 3
seconds to 10 seconds when placed in the mouth, are bioequivalent,
exhibiting peak concentrations at about 6 hours after
administration. Similarly, the first-pass effect in the liver
eliminates approximately 40% of the dose before reaching systemic
circulation.
[0240] In the present example, fast-dispersing tablets are prepared
by lyophilization as described above in this Example containing 10
mg olanzapine. Upon administration of the fast dispersing
olanzapine tablet by placing it in contact with buccal tissue, it
has been discovered that addition of certain alkylsaccharides
having specific alkyl chain lengths to the fast-dispersing
olanzapine tablets results in substantially reduced first-pass
effect metabolism of olanzapine as seen by a reduction in the
relative proportion of olanzapine metabolites in systemic
circulation compared to un-metabolized active drug. The relative
proportions of olanzapine and olanzapine metabolites in serum or
plasma can be determined using an HPLC Chromatograph, Perkin Elmer
200, with a Refractive Index Detector equipped with a thermostated
cell. A suitable solid-phase absorbent may be used such as
Lichrosorb RP-18 (Merck, Darmstadt, Germany) 250 mm, with a mobile
phase consisting of acetonitrile:water gradient. Injection volumes
of 20 .mu.L using the Perkin Elmer 200 auto-sampler and a flow rate
of 0.8 mL/minute are satisfactory for this purpose. Specifically,
incorporation of from 0.2% up to 10% dodecyl maltoside or
tetradecyl maltoside or sucrose dodecanoate in a fast-dispersing
tablet format increases the drug that enters into systemic
circulation and decreases the drug that is eliminated by the
"first-pass" effect in the liver. Additionally, the time to maximum
drug levels is dramatically reduced, typically from one to six
hours, to as little as approximately 15 to 45 minutes. For use in
treating combative patients undergoing psychotic episodes, this
more rapid absorption of drug, resulting in more rapid onset of
action, may be of great benefit.
EXAMPLE 11
Preparation of Fast-Dispersing Dosage Forms of Melatonin
[0241] Melatonin or 5-methoxy-N-acetyltryptamine is a neurohormone
used to regulate sleep-wake cycles in patients with sleep
disorders. Endogenous melatonin is secreted by the pineal gland in
all animals exhibiting circadian or circannual rhythms. Melatonin
plays a proven role in maintaining sleep-wake rhythms, and
supplementation may help to regulate sleep disturbances that occur
with jet lag, rotating shift-work, depression, and various
neurological disabilities.
[0242] Commercially available formulations of melatonin include
oral and sublingual tablets, capsules, teas, lozenges, and oral
spray delivery systems. Oral melatonin administration follows a
different pharmacokinetic profile than that of the endogenous
hormone. After oral administration, melatonin undergoes significant
first-pass hepatic metabolism to 6-sulfaoxymelatonin, producing a
melatonin bioavailability estimated at 30-50%. DeMuro et al. (2000)
reported that the absolute bioavailability of oral melatonin
tablets studied in normal healthy volunteers is somewhat lower at
approximately 15%. The mean elimination half-life of melatonin is
roughly 45 minutes.
[0243] Fast-dispersing melatonin tablets are prepared containing 1
mg, 5 mg, 10 mg and 20 mg according to the method described in
Example 10 above, with and without 1% to 2% alkylsaccharide as
described in Example 10. New Zealand White rabbits are anesthetized
are placed into a restraining box and anesthetized using a single
administration of acepromazine/ketamine (0.7 mg/0.03 mg in 0.1 mL)
administered by injection into the marginal ear vein) to facilitate
dosing. This results in anesthesia for a period of about 10 minutes
during which time the animals are dosed with test article.
Thereafter, the animals return to consciousness. At individual time
point over a two hour period, 1 mL blood samples are collected from
the central ear artery. After collection, plasma is immediately
prepared from each blood sample using lithium/heparin as the
anticoagulant. All samples are stored at -70.degree. C. until
assaying for melatonin. Melatonin is measured using a commercial
ELISA kit manufactured by GenWay Biotech Inc., San Diego, Calif.
Upon administration by contacting the fast-disintegrating tablets
with buccal tissue in the upper portion of the mouth, melatonin is
found to be absorbed with a bioavailability of at least 75% as
measured by area under the curve in the presence of alkylsaccharide
and less than 50% in the absence of alkylsaccharide. Melatonin is
measured using a commercial ELISA kit (No. 40-371-25005)
manufactured by GenWay Biotech Inc., San Diego, Calif. In addition,
for the tablets containing alkylsaccharide the maximal
concentration of melatonin is reached in approximately one half the
time it takes for tablets not containing alkyl saccharides.
EXAMPLE 12
Preparation of Fast-Dispersing Dosage Forms of Raloxifene
[0244] Raloxifene, also called Evista.RTM. is used for the
treatment and prevention of osteoporosis in postmenopausal women,
the reduction in the risk of invasive breast cancer in
postmenopausal women with osteoporosis, and the reduction in the
risk of invasive breast cancer in postmenopausal women at high risk
of invasive breast cancer. The recommended dosage is one 60 mg
tablet daily. While approximately 60% of an oral dose of raloxifene
is absorbed rapidly after oral administration, presystemic
glucuronide conjugation is extensive, resulting in an absolute
bioavailability for raloxifene of only 2%. A fast-dispersing 60 mg
raloxifene tablets prepared as described in U.S. Pat. No. 5,576,014
or 6,696,085 B2 or 6,024,981 are found to have very similar
pharmacokinetics with approximately 2% absolute bioavailability.
However, a fast-dispersing tablet containing 10 mg or less of
micronized raloxifene-prepared by spray-dried dispersion (Bend
Research Inc., Bend Oreg., or AzoPharma, Miramar, Fla.) or by more
commonly used standard pharmaceutical grinding or milling
processes, and 0.5% to 5% dodecyl maltoside, when administered
buccally achieves systemic drug levels similar to those achieved
with the 60 mg oral tablet and at the same time results in less
circulating inactive raloxifene glucuronide.
[0245] While clinical benefit results primarily from the
unconjugated drug, side effects may be mediated by either or both
active drug and substantially inactive glucuronide conjugated drug.
Thus reducing exposure to the inactive drug conjugate, in this case
present in as much as a 30-fold higher concentration than active
drug, affords potentially significant clinical benefit in reducing
the likelihood of side effects. Raloxifene has a water solubility
of approx. 0.25 mg/L. As a result, it is not possible to dissolve
raloxifene in water in preparation for lyophilization to prepare a
fast-dispersing formulation as described in Example 10.
[0246] In this case, a self-assembling hydrogel can be formed by
adding 1% to 30% w/w CRODESTA F-110 in a suitable buffer, which is
vortexed and heated to 45 degrees for 1 hr. Then raloxifene in a
fine particle or micronized form is added to the warm liquid to
achieve a concentration in suspension of 60 mg/mL which is again
mixed by vortexing until the solid is uniformly suspended and
dispersed. Upon cooling to room temperature, a stable thixotropic
hydrogel forms which is capable of being dispensed but which
maintains the uniform suspension. Acetate buffer in the pH range of
pH 2 to pH 7 is found to be particularly well suited for this
purpose. Aliquots of 1 mL of the gel suspension of raloxifene are
placed in the wells of a 24 well disposable microwell plastic plate
and lyophilized as described in Example 1.
[0247] Administration of this fast dispersing formulation upon
presentation to buccal tissue results in an increase (a doubling)
in absolute bioavailability to at least 4% and a corresponding
measurable reduction in the ratio of circulating raloxifene
glucuronide conjugate concentration to unconjugated raloxifene.
EXAMPLE 13
Preparation of Fast-Dispersing Dosage Forms of Diphenhydramine
[0248] Diphenhydramine is a sedating antihistamine with pronounced
central sedative properties and is used as a hypnotic in the
short-term management of insomnia, symptomatic relief of allergic
conditions including urticaria and angioedema, rhinitis and
conjunctivitis, pruritic skin disorders, nausea and vomiting,
prevention and treatment of motion sickness, vertigo, involuntary
movements due to the side effects of certain psychiatric drugs and
in the control of parkinsonism due to its antimuscarinic
properties. A particularly desirable characteristic of
diphenhydramine is its apparent lack of any evidence of creating
dependency. Because of its excellent safety profile, it is
available as an over-the-counter drug and unlike some of the newer
sleep medications such as Ambien.RTM. and Lunesta.RTM. which can
cause bizarre behaviors such as sleepwalking and eating-binges
while asleep, along with occasional severe allergic reactions and
facial swelling causing the FDA to require label warnings about
these side effects for these newer prescription medications.
[0249] Diphenhydramine hydrochloride is given by mouth in usual
doses of 25 to 50 mg three or four times daily. The maximum dose in
adults and children is about 300 mg daily. A dose of 20 to 50 mg
may be used as a hypnotic in adults and children over 12 years old.
The drug is well absorbed from the gastrointestinal tract; however
it is subject to high first-pass metabolism which appears to affect
systemic drug levels. Peak plasma concentrations are achieved about
1 to 4 hours after oral doses. Diphenhydramine is widely
distributed throughout the body including the CNS and due to its
extensive metabolism in the liver, the drug is excreted mainly in
the urine as metabolites with small amounts of unchanged drug found
to be present.
[0250] While diphenhydramine is considered safe and effective for
treatment of insomnia and other disorders, the relatively long
onset of action due to the delay in achievement of peak plasma
concentrations of from one to four hours is inconvenient and
reduces the practical utility of this safe and effective drug.
Intravenously administered diphenhydramine exerts a rapid onset of
action; however, intravenous administration is not practical for
outpatient use or non-serious medical indications. The need for a
rapid onset-of-action formulation of diphenhydramine is clear. In
the case of insomnia, a patient may need to take the current oral
forms of the drug well in advance of going to bed in order to
minimize the likelihood of extended restless sleeplessness while
waiting for the drug to achieve sufficient systemic drug levels in
order to exert its desired pharmacological effect. In the case of
the antiemetic applications of diphenhydramine, rapid onset of
action is also highly desirable in order to relieve nausea and
vomiting as soon as quickly as possible. This is likewise the case
in the treatment of motion sickness and vertigo since these
symptoms can arise unexpectedly and it is both inconvenient and
undesirable to have to wait one to four hours while the orally
administered drug achieves sufficient systemic drug levels to
achieve its beneficial effects.
[0251] Diphenhydramine has a solubility in water of approximately
3.06 mg/mL. Therefore the method described in Example 12 may be
used to prepare fast-dispersing diphenhydramine tablet containing
50 mg of drug and 1% to 2% alkylsaccharide. Because Diphenhydramine
is slightly bitter, a taste masking amount of a pharmaceutically
acceptable flavor and a sweetener may be added to improve
palatability. Fast-dispersing tablets prepared in this manner have
a more rapid onset of action compared to "whole-swallowed" tablets
syrup, chewable tablets, lozenge, or edible film-strip and exhibit
less first-pass metabolism as well.
EXAMPLE 14
Administration of Alkylglycosides with Anti-Obesity Peptide Mouse
[D-Leu-4]OB3 to Mice
[0252] This example shows the uptake of anti-obesity peptide mouse
[D-Leu-4]OB3 in 0.3% alkylglycoside tetradecyl-beta-D-maltoside
(Intravail.TM. A3) by male Swiss Webster Mice. The synthetic leptin
agonist [D-Leu-4]OB3 mixed with 0.3% alkylglycoside
tetradecyl-beta-D-maltoside (Intravail.TM. A3), was administered to
six-week old male Swiss Webster mice at a dose of 1 mg by
gavage.
[0253] Mouse [D-Leu-4]OB3 (at a concentration of 1 mg/200 ul) was
dissolved in either PBS (pH 7.2) or 0.3% alkylglycoside
tetradecyl-beta-D-maltoside (Intravail.TM. A3) reconstituted in PBS
(pH 7.2) and administered by gavage, without anesthesia, to each of
4 mice per time point. After 10, 30, 50, 70, 90 or 120 minutes, the
mice were euthanized by inhalation of isoflurane (5%) and
exsanguinated by puncture of the caudal vena cava. Blood was also
collected from four mice not given peptide (prebleed). The blood
from each of the four mice in the time period was pooled, and serum
samples were prepared. Mouse [D-Leu-4]OB3 content of the pooled
samples was measured by competitive ELISA.
[0254] These experiments were repeated twice. The data collected
from a single experiment are presented in Table X and FIG. 4. The
data were determined to be highly reproducible. Uptake curves were
plotted using Microsoft.TM. Excel, and AUC was calculated using a
function of the graphics program SigmaPlot 8.0.TM. (SPSS Science,
Chicago, Ill.). The lowest AUC value obtained was arbitrarily set
at 1.0. Relative bioavailability was determined by comparing all
other AUC values to 1.0.
TABLE-US-00010 TABLE X Uptake of 1 Mg Mouse p-Leu-4]OB3 in 0.3%
Alkylglycoside Tetradecyl-beta-D-maltoside (Intravail .TM. A3) By
Male Swiss Webster Mice Following Administration By Gavage Relative
Sample AUC bioavailability Mouse [D-Leu-4]OB3 137,585 ng/ml/min 1.0
in PBS Mouse [D-Leu-4]OB3 552,710 ng/ml/min 4.0 in 0.3%
alkylglycoside tetradecyl-beta-D- maltoside (Intravail .TM. A3)
[0255] As evidenced in Table X and FIG. 4, addition of
alkylglycoside tetradecyl-beta-D-maltoside (Intravail.TM. A3) at
0.3% increases relative absorption of the OB-3 peptide by 4-fold
compared to peptide in PBS alone.
EXAMPLE 15
Administration of Alkylglycosides with Sumatriptan to Canines
[0256] This example shows the uptake of sumatriptan in 0.5%
alkylglycoside tetradecyl-beta-D-maltoside (Intravail.TM. A3) by
canines. Sumatriptan mixed with 0.5% alkylglycoside
tetradecyl-beta-D-maltoside (Intravail.TM. A3), was administered to
canines as a dose of 25 mg by both oral and rectal
administration.
[0257] As evidenced in FIG. 5, addition of alkylglycoside
tetradecyl-beta-D-maltoside (Intravail.TM. A3) at 0.5% increases
C.sub.max of sumatriptan for both oral and rectal administration as
compared to currently available 25 mg oral tablets. C.sub.max for
currently available tablets was determined to be 104 ng/ml for
canines as represented by the horizontal dashed line in FIG. 5.
EXAMPLE 16
Oral Administration of Octreotide
[0258] Octreotide in three oral concentrations of
n-dodecyl-beta-D-maltoside (DDM) (0.5%, 1.5%, and 3% DDM) and a
subcutaneous injection (s.c.) of octreotide in buffer (s.c.
Octreotide) containing no Intravail.RTM. is administered to four
respective groups of 24 mice each. The animal test groups are
described below in Table 2. Dosing solutions may be stored
refrigerated at 4-8 deg. C prior to administration. The oral and
subcutaneous doses administered are adjusted to be 1000 .mu.g/kg
average body mass/group (which is 30 .mu.g for 30 g mice),
administered by 200 .mu.L oral gavage or subcutaneously as a 100 uL
injection between the skin and underlying tissue layers in the
scapular region on the back of each animal. Mice are anesthetized
with 5% isoflurane, and blood is collected by cardiac puncture over
a three hour time period at 0, 5, 10, 15, 30, 60, 120 and 180
minutes immediately prior to or following either oral or
subcutaneous administration of octreotide. Death is confirmed by
cervical dislocation. After blood collection, serum is immediately
prepared from each blood sample. Dosing solutions and all serum
samples are stored at -70.degree. C. until assayed as described
below. Collection of blood and serum preparation--5, 10, 15, 30,
60, 120 or 180 min after octreotide delivery, the mice (n=six per
time point) are anesthetized with isoflurane (5%) and exsanguinated
by cardiac puncture. Euthanasia is confirmed by cervical
dislocation. The blood is collected in sterile nonheparinized
plastic centrifuge tubes and allowed to stand at room temperature
for 1 h. The clotted blood is rimmed from the walls of the tubes
with sterile wooden applicator sticks. Individual serum samples are
prepared by centrifugation for 30 min at 2600.times.g in an
Eppendorf 5702R, A-4-38 rotor (Eppendorf North America, Westbury,
N.Y., USA), The serum samples in each experimental group are pooled
and stored frozen until assayed for octreotide content by EIA. The
three treatment groups and s.c. control group are as follows: 1)
oral Octreotide, 0.5% DDM; 2) oral Octreotide 1.5% DDM; 3) oral
Octreotide, 3.0% DDM; and 4) s.c. Octreotide. At time zero (0),
octreotide is delivered subcutaneously or by gavage to each mouse.
Following treatment, the mice are transferred to separate cages for
the designated time period.
[0259] The Animal Test System that was used in these studies is
described in Table XI below. Animals are segregated by weight into
each of the four treatment groups to minimize variation within
groups. The animals are housed individually in polycarbonate cages
fitted with stainless steel wire lids and air filters and supported
on ventilated racks (Thoren Caging Systems, Hazelton, Pa., USA).
The mice are maintained at a constant temperature (24.degree. C.)
with lights on from 07:00 to 19:00 hours and allowed food and water
ad libitum.
TABLE-US-00011 TABLE XI Animal Test System Species (strain): Male
Swiss Webster (SW-M) mice, 6 to 7 weeks of age Supplier: Taconic
Farms # of males: Four groups of 24 mice (96 total) # of females 0
Age: 6 to 7 weeks of age Housing: Three animals in plastic shoebox
cages Food: Rodent Chow Availability of water: Ad lib Availability
of food: Ad lib
[0260] Octreotide is obtained from BCN (Spain) or Polypeptide
Laboratories (California, USA). Octreotide stock solutions are
prepared as described in Table XII by dissolving the lyophilized
powder in pH 4.5 acetate buffer 0.1% EDTA (Table 2) containing 0.0%
DDM (s.c. control), 0.5%, 1.5% or 3% DDM. The appropriate dose is
administered to animals in each group as listed in Table XIII. All
of these animal procedures are reviewed and approved by the
institutional Animal Care and Use Committee, and are performed in
accordance with relevant guidelines and regulations. The dosing
solution remaining after administration is divided and frozen at
-70.degree. C. until assayed.
TABLE-US-00012 TABLE XII pH 4.5 mM Sodium Acetate Buffer, 0.1% EDTA
Component Quantity Acetic acid 0.286 mL 1N NaOH adjust to pH 4.5
Na2 EDTA 500 mg water 500 mL Adjust pH: pH 4.5
TABLE-US-00013 TABLE XIII Dosing Solutions in pH 4.5 Acetate
Buffer, 0.1% EDTA & Dose Administration Total Final Dose DDM
Octreotide Octreotide Volume (30 g Group (mg/5 mL) (mg)*
Concentration Administered mouse) Oral 50 mg in plus 1.5 mg in 150
ug/mL 200 .mu.L 30 .mu.g Octreotide- 5 mL 5 mL (10 mL total 0.5% A3
vol.) Oral 150 mg in plus 1.5 mg in 150 ug 200 .mu.L 30 .mu.g
Octreotide- 5 mL 5 mL (10 mL total 1.5% A3 vol.)/mL Oral 300 mg in
plus 1.5 mg in 150 ug/mL 200 .mu.L 30 .mu.g Octreotide- 5 mL 5 mL
(10 mL total 3% A3 vol.) s.c. N/A N/A 1.5 mg in 300 ug/mL 100 .mu.L
30 .mu.g Octreotide 5 mL (5 mL total vol.) *Prepared as 6 mg
dissolved in 20 mL acetate buffer
[0261] Octreotide concentrations for dosing solution(s) and pooled
serum samples for each time period for each treatment group are
assayed in triplicate using an octreotide enzyme immunoassay assay
(EIA) (Peninsula Laboratories, LLC (San Carlos, Calif.) Cat. No.
S-1342--Octreotide for Serum and Plasma Samples) according to the
instructions supplied by the manufacturer.
[0262] Pharmacokinetic analyses is carried out as follows. To
determine relative bioavailability, serum concentrations of
octreotide vs. time following s.c. and oral delivery are plotted
using the graphics program SigmaPlot.TM. 8.0 (SPSS Science,
Chicago, Ill., USA). The area under each curve (AUC) is calculated
with a function of this program. The lowest AUC value obtained is
arbitrarily set at 1.0. Relative bioavailability is determined by
comparing all other AUC values to 1.0.
[0263] Serum half-life (t.sub.1/2) is determined as follows. The
period of time required for the serum concentration of octreotide
to be reduced to exactly one-half of the maximum concentration
achieved following s.c. or oral administration is calculated using
the following formula:
t.sub.1/2=0.693/k.sub.elim
[0264] k.sub.elim represents the elimination constant, determined
by plotting the natural log of each of the concentration points in
the beta phase of the uptake profiles against time. Linear
regression analysis of these plots results in straight lines, the
slope of which correlates to the k.sub.elim for each delivery
method.
[0265] Clearance of octreotide from the plasma following s.c. or
oral delivery is calculated from the AUC using the following
equation:
CL=Dose/AUC
[0266] Since the half-life of a drug is inversely related to its
clearance from the plasma and directly proportional to its volume
of distribution, the apparent volume of distribution of octreotide
following s.c. or oral delivery is calculated from its half-life
and clearance using the following equation:
t.sub.1/2=(0:693.times.V.sub.d)/CL
[0267] Results: Octreotide uptake profiles following s.c. and oral
delivery in 0.5%, 1.5% or 3.0% Intravail.RTM. are shown in FIGS. 6
to 9, respectively. All of these profiles show biphasic uptake of
octreotide with an initial peak (C.sub.max1) at 10 min (t.sub.max1)
followed by a second peak (C.sub.max2) at 30 min (t.sub.max2).
C.sub.max1 and C.sub.max2 are approximately the same (6.67 ng/ml
vs. 7.59 ng/ml, respectively) following s.c. administration, and
decrease at different rates after each of the two peaks (FIG.
6).
[0268] Oral delivery of octreotide in 0.5% Intravail.RTM. produces
an uptake profile (FIG. 7) with a C.sub.max1 more than 2-fold
higher than C.sub.max2 (59.7 ng/ml vs. 25.9 ng/ml, respectively).
When the Intravail.RTM. concentration is increased to 1.5% or 3.0%,
C.sub.max1 is reduced to 17.8 ng/ml and 3.75 ng/ml, respectively.
Likewise 1.5% or 3.0% Intravail.RTM. also reduces C.sub.max2 to 4.0
ng/ml and 2.48 ng/ml, respectively. As observed after s.c.
delivery, octreotide concentrations following oral delivery in
0.5%, 1.5% or 3.0% Intravail.RTM. decrease at different rates after
each of the two peaks.
[0269] The relative bioavailability of octreotide is determined by
measuring the area under the uptake curve (AUC) for each delivery
method. This value represents the total extent of peptide
absorption into the systemic circulation, or total uptake,
following its administration. Because of the biphasic nature of the
uptake profiles, the relative bioavailability of octreotide
following s.c. and oral delivery is determined by measuring the AUC
for each of the two peaks in the profile separately, and determined
as follows: AUC=AUC.sub.1+AUC.sub.2. Using this formula, the AUC of
octreotide after s.c. administration is determined to be 290
ng/ml/min, and assigned a relative bioavailability of 1.0. The AUC
of octreotide following oral delivery in 0.5%, 1.5% or 3.0%
Intravail.RTM. is 1,254 ng/ml/min, 230.7 ng/ml/min, and 141.24
ng/ml, respectively, and is assigned relative bioavailabilities of
4.3, 0.8 and 0.6.
[0270] To determine the serum half-life of octreotide following
s.c. and oral delivery, the k.sub.elim for each peak in the uptake
curves is calculated separately (k.sub.elim1 and k.sub.elim2).
These values are then used to determine the half-life of octreotide
under each peak (t.sub.1/2 1 and t.sub.1/2 2). The overall
half-life is calculated as follows: t.sub.1/2=t.sub.1/2 1+t.sub.1/2
2, and is determined to be 41.2 min following s.c. delivery and
53.1 min, 25.8 min and 23.6 min following oral delivery in 0.5%,
1.5% or 3.0% Intravail.RTM., respectively.
[0271] Because of the biphasic profile of the uptake curves
associated with s.c. and oral delivery of octreotide, plasma CL is
measured using the AUC associated with each peak in the profile:
CL.sub.1=Dose/AUC.sub.1 and CL.sub.2=Dose/AUC.sub.2. Overall
clearance is calculated as follows: CL=CL.sub.1+CL.sub.2, and is
determined to be 30 L/min following s.c. administration, and 3.9
L/min, 28.4 L/min and 54.9 L/min following oral delivery in 0.5%,
1.5% or 3.0% Intravail.RTM., respectively.
[0272] The apparent volume of distribution (Vd) of octreotide
following s.c. and oral delivery is calculated using the half-life
and clearance rate determined for each peak associated with the
biphasic uptake profiles: t.sub.1/2 1=0.693V.sub.d1/CL.sub.1 and
t.sub.1/2 2=0.693.times.V.sub.d2/CL.sub.2. The overall apparent
volume of distribution is calculated as follows:
V.sub.d=V.sub.d1+V.sub.d2 and determined to be 301.1 L following
s.c. delivery and 84.7 L, 299.3 L and 357.8 L following oral
delivery in 0.5%, 1.5% or 3.0% Intravail.RTM..
[0273] When the pharmacokinetics of orally delivered (by gavage)
octreotide in increasing concentrations (0.5%, 1.5% and 3.0%) of
DDM are compared to the pharmacokinetics of octreotide delivered
subcutaneously, oral delivery of octreotide in 0.5% DDM is seen to
significantly enhanced total uptake (1,254.08 ng/ml/min vs. 290.12
ng/ml/min, respectively) and relative bioavailability (4.3 vs. 1.0,
respectively) when compared to delivery by s.c. injection. Higher
concentrations of DDM do not further enhance uptake or
bioavailability. The half-life of octreotide is increased by oral
delivery in 0.5% DDM from 41.2 min (s.c.) to 52.1 min, and
clearance from the plasma is reduced from 30.0 L/min (s.c.) to 3.9
L/min. The results indicate that oral delivery of octreotide in
compositions containing DDM is feasible, and is an effective method
of administration for achieving high serum levels of octreotide
when compared to s.c. injection. All pharmacokinetic parameters
measured in this study are summarized in Table XIV. In addition to
DDM, n-tetradecyl maltoside, n-tridecyl maltoside and sucrose
monododecanoate may be used to substitute for DDM to get similar
results.
TABLE-US-00014 TABLE XIV Pharmacokinetic Parameters of Octreotide
Uptake in Male Swiss Webster Mice Following Subcutaneous Delivery
in 10 mM Sodium Acetate Buffer Containing 0.1% EDTA (pH 4.5) or
Oral Administration (by gavage) in Increasing Concentrations of
DDM. Oral Oral Oral S.C. 0.5% DDM 1.5% DDM 3.0% DDM Cmax (ng/ml)
Cmax1 6.67 59.68 17.8 3.75 Cmax2 7.59 25.92 4.0 2.48 Tmax (min)
Tmax1 10 10 10 10 Tmax2 30 30 30 30 AUC (ng/ml/min) 290.12 1254.08
230.70 141.24 AUC.sub.1 38.39 353.03 97.80 21.50 AUC.sub.2 251.73
901.04 132.90 119.74 Relative bioavailability 1.0 4.3 0.8 0.6 kelim
(ml/min) k.sub.eliml 0.3400 0.4400 0.7200 0.5200 k.sub.elim2 0.0177
0.0132 0.0279 0.0311 t.sub.1/2 (min) 41.2 52.1 25.8 23.6 t 1/2 1
2.04 1.56 0.96 1.33 t 1/2 2 39.15 52.50 24.84 22.28 CL (L/min) 30.0
3.9 28.4 54.9 CL.sub.1 26.0 2.8 20.9 46.5 CL.sub.2 4.0 1.1 7.5 8.4
Vd (L) 301.1 84.7 299.3 357.8 Vd.sub.1 76.7 0.6 29.0 89.3 Vd.sub.2
224.4 84.1 270.3 268.5
EXAMPLE 17
Oral Administration of the GLP-1 Analog, Liraglutide
[0274] Objective: Test oral bioavailability of liraglutide with
Intravail.TM. A3 during a challenge with dietary sugar
[0275] Procedure was as follows. 500 uL of liraglutide was
extracted from pens and placed in 50 mm test tube. 50 uL 5% A3 was
added in H2O--0.45% final A3 concentration. Composition was mixed
with disposable plastic dropper. 40 grams of dietary sugar was
ingested and 60 minutes allowed to pass. Blood glucose was measured
(upper arm just above elbow on left arm) using meter=T.sub.o.
Deposit composition on back of tongue and swallow. Blood glucose
level was measured and recorded at T=10, 20, 30, 40, 55, 70, 80 and
90 min.
[0276] Results: Blood glucose declined from 146 mg/dL down to 100
mg/dL at 80 min. The rate of decline in he presence of a dietary
sugar challenge was slower than in the non-challenged state
observed previously.
[0277] Conclusion: Oral liraglutide using Intravail A3 at 0.45%
effectively increases bioavailability and delivers liraglutide to
the blood stream. The 500 uL dose was 1.4.times. the standard
injectable dose and the PD effect was large, so bioavailability is
substantial--approximately 30% to 60%. This confirms previously
observed high relative bioavailability as compared to delivery
without alkylglycoside. Effectiveness under conditions of dietary
sugar challenge is also demonstrated. Blood glucose levels are
shown in FIG. 10 while the dose response is shown in FIG. 11. A
positive dose response is seen further confirming the
pharmacodynamics.
TABLE-US-00015 TABLE XV Effect of Orally Delivered Liraglutide on
Blood Glucose Values Liraglutide Oral Data glucose (mg/dL) time
Trial 1 Trial 2 -20 125 -10 144 0 116 146 10 122 127 20 102 124 30
101 128 40 104 115 55 111 70 108 80 100 90 105
EXAMPLE 18
***Oral Delivery of Phenylephrine
[0278] In a canine model, subjects were dosed orally with various
phenylephrine (PE) tablet formulations, and blood samples were
taken to determine the rate of parent phenyl ethyl phenylephrine
absorption. Blood levels of parent phenylephrine were measured.
Three subject groups were created with each alkylsaccharide test
group containing five subjects in each test group. All subject
groups received 10 mg of phenylephrine as a single tablet. Each
subject was in the fasting state for overnight and two hours post
dose. Blood samples were taken at pre-dose, 5, 10, 15, 30, 45 min.,
1, 2, and 3 hours.
[0279] The tablet formulations tested are shown in Table XVI.
Comparator was the standard 30 mg (total weight including
excipients) phenylephrine tablet was no absorption enhancer. Group
number 2 tablets contain the alkylsaccharide designated A3, which
is n-dodecyl-beta-D-maltoside in the amounts of 2.5 mg, 5 mg, and
7.5 mg, respectively. Group number three tablets contain the
alkylsaccharide designated B3, which is sucrose monododecanoate, in
the amounts of 5 mg, 10 mg, and 20 mg, respectively.
TABLE-US-00016 TABLE XVI Comparator (1) Regular 10 mg PE Aegs A3
(2) 10 mg PE + 2.5 mg A3 10 mg PE + 5 mg A3 10 mg PE + 7.5 mg A3
Aegs B3 (3) 10 mg PE + 5 mg B3 10 mg PE + 10 mg B3 10 mg PE + 20 mg
B3
TABLE-US-00017 TABLE XVII AUC AUC (0-3 hour) Test Articles (0-3
hour) Change Ratio 2.5 mg A3 + 10 mg PE tablet 2229610 1.14 5 mg A3
+ 10 mg PE tablet 3055140 1.56 7.5 mg A3 + 10 mg PE tablet 1648846
0.84
TABLE-US-00018 TABLE XVIII AUC AUC (0-3 hour) Test Articles (0-3
hour) Change Ratio 5 mg B3 + 10 mg PE tablet 1684238 0.86 10 mg B3
+ 10 mg PE tablet 2932360 1.50 20 mg B3 + 10 mg PE tablet 2333966
1.19
[0280] Results: As can be seen from FIG. 12 and Table XVII,
n-dodecyl-beta-D-maltoside increased oral bioavailability of
phenylephrine in compressed tablets by up to 56%. Similarly, as can
be seen from FIG. 13 and Table XVIII sucrose monododecanoate
increased oral bioavailability of phenylephrine in compressed
tablets by up to 50%. Interestingly, and unexpectedly, the maximum
increase in bioavailability occurred at intermediate levels,
between the low and high alkylsaccharide levels tested. In the case
of n-dodecyl-beta-D-maltoside, this occurred at a loading a 5 mg of
alkylsaccharide. Whereas in the case of sucrose mono-dodecanoate
this occurred at a loading of 10 mg of alkylsaccharide. However, a
lower alkylsaccharide loading of dodecyl-beta-D-maltoside also
showed an increase over the comparator, whereas in the case of
sucrose mono-dodecanoate, a slightly higher alkylsaccharide loading
gave rise to an increase in bioavailability over the comparator.
The loading of alkylsaccharides in both cases has bracketed the
relative amounts useful in providing maximum bioavailability
enhancement. The actual amounts may vary as the amount of drug
varies and as the choice of drug may also vary. Thus, similar
formulation studies may be conducted with other drugs intended to
be formulated in solid dosage forms having enhanced
bioavailability, such that the relative loadings of alkylsaccharide
for the desired drug dose can be individually determined. Other
drugs for which these formulations and methods of testing are
useful include by way of example phenylephrine (as HCl or
bitartrate), aspirin, naproxen sodium, brompheniramine (maleate),
triprolidine, chlorpheniramine (maleate), dextromethorphan (HBr),
guaifenesin, acetaminophen, pseudaphedrine, epinephrine,
diphenhydramine, cimetidine, loratadine, ranitidine, famotidine,
ketoprofen, omeprazole, clemastine, dimenhydrinate, ibuprofen,
cyclizine (Marizine) or other pharmaceutically acceptable salts
thereof such as hydrochlorides, maleates, tartrates, acetates, and
the like, as individual (monotherapy) drugs or as combinations of
two or more drug substances in a solid dosage form.
[0281] Throughout this application, various publications are
referenced. The disclosures of these publication in their
entireties are hereby incorporated by reference into this
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[0307] Although the present process has been described with
reference to specific details of certain embodiments thereof in the
above examples, it will be understood that modifications and
variations are encompassed within the spirit and scope of the
invention. Accordingly, the invention is limited only by the
following claims.
* * * * *