U.S. patent application number 14/589811 was filed with the patent office on 2015-07-09 for formulations for delivering insulin.
The applicant listed for this patent is Emisphere Technologies, Inc.. Invention is credited to William E. Bay, Nikhil Dhoot, Steven Dinh, Halina LEVCHIK, Jun Liao, Puchun Liu, Shingai Majuru, Moses Oyewumi.
Application Number | 20150190344 14/589811 |
Document ID | / |
Family ID | 38610388 |
Filed Date | 2015-07-09 |
United States Patent
Application |
20150190344 |
Kind Code |
A1 |
LEVCHIK; Halina ; et
al. |
July 9, 2015 |
FORMULATIONS FOR DELIVERING INSULIN
Abstract
Oral insulin formulations and processes for preparing oral
insulin formulations are provided.
Inventors: |
LEVCHIK; Halina; (Croton On
Hudson, NY) ; Oyewumi; Moses; (Yorktown Heights,
NY) ; Majuru; Shingai; (Brewster, NY) ; Bay;
William E.; (Ridgefield, CT) ; Liao; Jun;
(Cedar Knolls, NJ) ; Liu; Puchun; (Chappaqua,
NY) ; Dinh; Steven; (Briacliff Manor, NY) ;
Dhoot; Nikhil; (Dombivli (east), IN) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Emisphere Technologies, Inc. |
Roseland |
NJ |
US |
|
|
Family ID: |
38610388 |
Appl. No.: |
14/589811 |
Filed: |
January 5, 2015 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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12297147 |
Oct 14, 2008 |
8927015 |
|
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PCT/US07/66560 |
Apr 12, 2007 |
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14589811 |
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60857747 |
Nov 7, 2006 |
|
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60791842 |
Apr 12, 2006 |
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Current U.S.
Class: |
424/452 ;
514/5.9 |
Current CPC
Class: |
A61K 9/2009 20130101;
A61K 9/2077 20130101; A61K 9/2095 20130101; A61K 9/145 20130101;
A61K 9/2027 20130101; A61K 9/2063 20130101; A61K 9/2013 20130101;
A61K 38/28 20130101; A61K 9/06 20130101 |
International
Class: |
A61K 9/20 20060101
A61K009/20; A61K 9/06 20060101 A61K009/06; A61K 38/28 20060101
A61K038/28 |
Claims
1-91. (canceled)
92. A tablet comprising (a) a delivery agent selected from
N-(8-[2-hydroxybenzoyl]amino) caprylic acid and pharmaceutically
acceptable salts thereof, (b) gelatin, (c) insulin, (d) dibasic
calcium phosphate, and (e) magnesium stearate, wherein the delivery
agent and insulin is in the form of a gel prior to being tableted
or filled in a capsule.
93. The tablet of claim 92, wherein the delivery agent is sodium
N-(8-[2-hydroxybenzoyl]-amino caprylate.
94. The tablet of claim 92, wherein the weight ratio of delivery
agent to insulin is about 45:1.
95. The tablet of claim 92, wherein the tablet consists of (a) the
delivery agent sodium N-(8-[2-hydroxybenzoyl]amino) caprylate, (b)
gelatin, (c) insulin, (d) dibasic calcium phosphate, and (e)
magnesium stearate, wherein the delivery agent and insulin is in
the form of a gel prior to being tableted or filled in a capsule,
and the weight ratio of delivery agent to insulin is about
45:1.
96. The tablet of claim 96, wherein the tablet consists of 240 mg
sodium N-(8-[2-hydroxybenzoyl]amino) caprylate, 12 mg gelatin, 5.45
mg insulin, 113.8 mg dibasic calcium phosphate, and 3.75 mg
magnesium stearate.
97. The tablet of claim 92, prepared by a process comprising (a)
mixing the delivery agent and the gelatin; (b) granulating the
product of step (a) with insulin; and (c) blending the granules of
step (b) with dibasic calcium phosphate and magnesium stearate.
98. A method of treating diabetes in a patient in need thereof
comprising administering to the patient a tablet of claim 92.
99. The method of claim 98, wherein the diabetes is Type II
diabetes.
100. The method of claim 98, wherein the patient does not exhibit
any statistically significant increase in weight, risk of
hypoglycemia or risk of hyperinsulinemia over the treatment
period.
101. A method of improving glucose tolerance and/or glycemic
control in a patient in need thereof comprising administering to
the patient a tablet of claim 92.
102. The method of claim 101, wherein the patient does not exhibit
any statistically significant increase in weight, risk of
hypoglycemia or risk of hyperinsulinemia over the treatment
period.
103. A solid dosage form comprising a delivery agent selected from
N-(8-[2-hydroxybenzoyl]amino) caprylic acid and pharmaceutically
acceptable salts thereof and insulin or insulin analog, wherein (i)
the solid dosage form is a tablet or capsule and (ii) (a) the
delivery agent and insulin are in the form of a gel or (ii) the
delivery agent and insulin or insulin analog are in the form of a
gel prior to being tableted or filled in a capsule.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a continuation of U.S. patent
application Ser. No. 12/297,147, filed Oct. 14, 2008, which is the
U.S. national phase of International Application No.
PCT/US07/66560, filed Apr. 12, 2007, which claims priority of
Provisional Application No. 60/791,842 filed Apr. 12, 2006 and to
Provisional Application No. 60/857,747 filed Nov. 7, 2006, the
specifications of both of which are herein incorporated by
reference.
FIELD OF THE INVENTION
[0002] The present invention relates to pharmaceutical formulations
(e.g. oral pharmaceutical formulations) containing insulin and one
or more delivery agents, e.g. (4-CNAB or sodium 4-CNAB), and
methods of treating subjects suffering from diabetes with such
pharmaceutical formulations.
BACKGROUND OF THE INVENTION
[0003] There is a need for orally administered insulin that
provides sufficient insulin bioavailability, and processes for
preparing such pharmaceutical compositions.
SUMMARY OF THE INVENTION
[0004] Methods of synthesizing pharmaceutical formulations
containing insulin and a delivery agent compound have an effect on
the bioavailability of the insulin upon administration of the
pharmaceutical formulation (e.g. upon oral administration by a
human). The present invention provides processing techniques that
facilitate the delivery of insulin upon administration with a
delivery agent compound.
[0005] One embodiment of the present invention is a solid oral
pharmaceutical composition comprising 4-CNAB, or a pharmaceutically
acceptable salt thereof, recombinant human insulin, povidone,
dibasic calcium phosphate, and magnesium stearate.
[0006] Another embodiment of the present invention is a process for
preparing a solid oral pharmaceutical formulation by introducing
insulin or an analog thereof to an aqueous solution that contains a
delivery agent compound, drying the solution to obtain an
insulin/delivery agent powder, optionally granulating the powder
with intragranular excipients, optionally adding extragranular
excipients to the powder or granules, and forming a unit dosage
form from the resulting composition (e.g., compressing the
composition into tablets or filling capsules with the
composition).
[0007] Another embodiment of the present invention is a process for
preparing a solid oral insulin pharmaceutical composition by (a)
preparing a solution of a delivery agent and insulin or an analog
thereof; (b) freeze-drying the insulin/delivery agent solution; (c)
milling the insulin/delivery agent colyophilized powder obtained by
freeze-drying the insulin/delivery agent solution; (d) mixing the
milled co-lyophilized powder with intragranular excipients; (e) dry
granulating the mixture formed in step (d), (f) adding
extragranular excipients; and (g) forming a unit dosage form from
the resulting composition (e.g., compressing the composition into
tablets or filling capsules with the composition). Dry granulation
may be performed, for example, by roller compaction or slugging and
then milling the resulting product.
[0008] Another embodiment of the present invention is a process for
preparing a solid oral insulin pharmaceutical composition by (a)
preparing a solution of a delivery agent and insulin or an analog
thereof; (b) performing rotary evaporation on the insulin/delivery
agent solution; and (c) forming a unit dosage form from the product
of step (b) (e.g., tableting the insulin/delivery agent powder or
adding the insulin/delivery agent powder to capsules). This process
may further include one or more of the steps of (d) milling the
insulin/delivery agent powder obtained by rotary evaporation; (e)
mixing the milled powder with intragranular excipients; (f)
granulating the milled powder and intragranular excipients (e.g. by
dry granulation), and (g) adding extragranular excipients. Dry
granulation may be performed, for example, by roller compaction or
slugging and milling.
[0009] Another embodiment of the present invention is a
pharmaceutical composition that includes (a) insulin or an insulin
analog, (b) a delivery agent and (c) a gelatin, as that term is
used herein (e.g. including gelatin alternatives).
[0010] Another embodiment of the present invention is a process for
preparing an oral insulin pharmaceutical composition by introducing
(a) a delivery agent compound and (b) insulin or an insulin analog
into gelatin or a gelatin alternative. In a preferred embodiment,
gelatin is optionally milled and mixed with a delivery agent. The
mixture is then granulated using an aqueous dispersion of insulin
or an insulin analog.
[0011] Another embodiment of the present invention is a method of
solubilizing insulin or an insulin analog by introducing a delivery
agent (e.g. 4-CNAB or sodium 4-CNAB) to an aqueous solution and
subsequently adding insulin or an insulin analog to the delivery
agent-containing solution. In one embodiment, the delivery agent is
added to water, sodium hydroxide is added to increase the pH of the
solution (e.g., to increase the pH to about 7 or 8), and then
insulin is added to the pH-adjusted solution.
[0012] Yet another embodiment is a method of treating diabetes in a
subject by administering a therapeutically effective amount of the
pharmaceutical composition of the present invention to the subject.
The diabetic subject can be a human suffering from Type I or Type
II diabetes. Generally, the insulin preparation of the present
invention does not induce any significant incidence of antibodies.
Preferably, the subject is administered the pharmaceutical
composition of the present invention for at least once day for at
least 90 days. According to a preferred embodiment, the
pharmaceutical composition is administered as adjunctive therapy to
a biguanide (such as metformin). According to another preferred
embodiment, the pharmaceutical composition is administered as
adjunctive therapy to a biguanide (such as metformin), acarbose, a
glitazone (e.g., pioglitazone), or a combination thereof.
[0013] One embodiment is a method of treating diabetes (e.g. type
II diabetes) in a subject who has not sufficiently responded to
metformin monotherapy, by administering a therapeutically effective
amount of the pharmaceutical composition of the present invention
to the subject. According to a preferred embodiment, the subject
continues treatment with metformin while also being treated with
the pharmaceutical composition of the present invention. According
to another preferred embodiment, the pharmaceutical composition is
administered as adjunctive therapy to a biguanide (such as
metformin), acarbose, a glitazone (e.g., pioglitazone), or a
combination thereof.
[0014] Yet another embodiment is a method of treating diabetes
(e.g., Type I or Type II diabetes) in a human having a hemoglobin
A1c value of at least about 8.0% by administering a therapeutically
effective amount of the pharmaceutical composition of the present
invention to the human. According to one embodiment, the human has
a hemoglobin A1c value ranging from about 8.0 to about 9.3%.
Preferably, the human is administered the pharmaceutical
composition of the present invention for at least once day for at
least 90 days.
BRIEF DESCRIPTION OF THE DRAWINGS
[0015] FIG. 1 sets forth the solubility of recombinant human zinc
insulin at different pH and time-points.
[0016] FIG. 2 sets forth a solubility curve for insulin and
4-CNAB.
[0017] FIGS. 3 and 4 set forth insulin dissolution profiles in
deionized water.
[0018] FIGS. 5-12 set forth results of insulin dosage forms
administered to monkeys as described in Example 7.
[0019] FIGS. 13 and 14 set forth the results of insulin dosage
forms administered to humans as described in Example 23.
[0020] FIG. 15 sets forth the results of insulin dosage forms
administered to humans as described in Example 24.
[0021] FIG. 16 sets forth the results of insulin dosage forms
administered to humans as described in Example 25.
[0022] FIGS. 17-24 sets forth the results of insulin dosage forms
administered to humans as described in Example 26.
DETAILED DESCRIPTION OF THE INVENTION
Definitions
[0023] As used herein and in the appended claims, the singular
forms "a" "an" and "the" also includes plural referents unless the
context clearly indicates otherwise. Thus, for example, reference
to "a molecule" includes one or more of such molecules, "a reagent"
includes one or more of such different reagents, reference to "an
antibody" includes one or more of such different antibodies, and
reference to "the method" includes reference to equivalent steps
and methods known to those of ordinary skill in the art that could
be modified or substituted for the methods described herein.
[0024] The term "hydrate" as used herein includes, but is not
limited to, (i) a substance containing water combined in molecular
form and (ii) a crystalline substance containing one or more
molecules of water of crystallization or a crystalline material
containing free water.
[0025] The term "solvate" as used herein includes, but is not
limited to, a molecular or ionic complex of molecules or ions of a
solvent with molecules or ions of the delivery agent compound or
salt thereof, or hydrate or solvate thereof.
[0026] The term "delivery agent" refers to any of the delivery
agent compounds disclosed or incorporated by reference herein.
[0027] The terms "alkyl", "alkoxy", "alkylene", "alkenylene",
"alkyl(arylene)", and "aryl(alkylene)" include, but are not limited
to, linear and branched alkyl, alkoxy, alkylene, alkenylene,
alkyl(arylene), and aryl(alkylene) groups, respectively.
[0028] Unless otherwise specified, the term "substituted" as used
herein includes, but is not limited to, substitution with any one
or any combination of the following substituents: halogens,
hydroxide, C.sub.1-C.sub.4 alkyl, and C.sub.1-C.sub.4 alkoxy.
[0029] The term "4-MOAC" refers to
8-(N-2-hydroxy-4-methoxybenzoyl)-aminocaprylic acid and
pharmaceutically acceptable salts thereof. Unless otherwise noted,
the term "4-MOAC" refers to all forms of 4-MOAC, including, but not
limited to, amorphous and crystalline forms of 4-MOAC.
[0030] The term "NAC" as used herein refers to
N-(8-[2-hydroxybenzoyl]-amino) caprylic acid and pharmaceutically
acceptable salts thereof, including its monosodium salt. Unless
otherwise noted, the term "NAC" refers to all forms of NAC,
including, but not limited to, all amorphous and crystalline forms
of NAC. The term "SNAC" as used herein refers to the monosodium
salt of NAC, including, but not limited to, all amorphous and
crystalline forms of SNAC (such as those described in International
Publication No. WO 2005/107462, which is hereby incorporated by
reference), unless otherwise indicated.
[0031] The term "NAD" as used herein refers to
N-(10-[2-hydroxybenzoyl]-amino) decanoic acid and pharmaceutically
acceptable salts thereof, including, but not limited to, its
monosodium salt. Unless otherwise noted, the term "NAD" refers to
all forms of NAD, including, but not limited to, all amorphous and
crystalline forms of NAD. The term "SNAD" as used herein refers to
the monosodium salt of NAD, including, but not limited to, all
amorphous and crystalline forms of SNAC.
[0032] The term "5-CNAC" refers to
N-(8-[2-hydroxy-5-chlorobenzoyl]-amino)octanoic acid (also known as
8-(N-2-hydroxy-5-chlorobenzoyl)aminocaprylic acid)) and
pharmaceutically acceptable salts thereof, including, but not
limited to, its monosodium salt and disodium salt. Unless otherwise
noted, the term "5-CNAC" refers to all forms of 5-CNAC, including,
but not limited to, all amorphous and crystalline forms of it
(including those described in International Publication No. WO
00/59863, PCT/US2006/036455, filed Sep. 18, 2006, and U.S.
Provisional Application No. 60/718,829, filed Sep. 19, 2005, all of
which are hereby incorporated by reference).
[0033] The term "4-CNAB" refers to
4-[(2-hydroxy-4-chlorobenzoyl)amino]butanoate (also known as
4-[(4-chloro-2-hydroxy-benzoyl)amino]butanoic acid) and
pharmaceutically acceptable salts thereof, including, but not
limited to, its monosodium salt. Unless otherwise noted, the term
"4-CNAB" refers to all forms of 4-CNAB, including, but not limited
to, all amorphous and crystalline forms of 4-CNAB. The term
"mono-sodium 4-CNAB" refers to monosodium
4-[(2-hydroxy-4-chlorobenzoyl)amino]butanoate, including, but not
limited to, anhydrous, monohydrate, and isopropanol solvates
thereof and amorphous and crystalline forms thereof (including
those described in International Publication Nos. WO 02/02509 and
WO 03/057650, both of which is hereby incorporated by reference),
unless otherwise indicated.
[0034] The term "HPOD" refers to
8-(2-hydroxyphenoxyl)octyldiethanolamine and pharmaceutically
acceptable salts thereof, including, but not limited to, its
meslyate salt. Unless otherwise noted, the term "HPOD" refers to
all forms of HPOD, including, but not limited to, all amorphous and
crystalline forms of HPOD and includes anhydrous, monohydrate, and
isopropanol solvates of HPOD, including those described in
International Publication No. WO 2005/115406, which is hereby
incorporated by reference).
[0035] The term "insulin analog" as used herein, refers to analogs
of naturally occurring insulins, including human insulin or animal
insulins, which differ by substitution of at least one naturally
occurring amino acid residue with other amino acid residues and/or
addition/removal of at least one amino acid residue from the
corresponding, otherwise identical, naturally occurring insulin.
Insulin analogs have a physiological effect similar to that of
naturally occurring or recombinant human insulin. The added and/or
replaced amino acid residues can also be those which do not occur
naturally. Insulin analogs include those analogs disclosed in U.S.
Pat. Nos. 6,960,561, 6,906,028, 6,852,694, 6,777,207, 6,630,348,
6,551,992, 6,534,288, 6,531,448, RE37,971, 6,465,426, 6,444,641,
6,335,316, 6,268,335, 6,051,551, 6,034,054, 5,970,973, 5,952,297,
5,922,675, 5,888,477, 5,873,358, 5,747,642, 5,693,609, 5,650,486,
5,646,242, 5,547,929, 5,504,188, 5,474,978, 5,461,031, 5,135,866,
4,421,685, all of which are hereby incorporated by reference.
[0036] The term "subject" as used herein includes a mammal,
preferably a human. It may also mean other animals, including other
mammals, but especially birds, poultry or other avian forms.
[0037] The phrase "pharmaceutically acceptable" refers to
components or compositions that are physiologically tolerable.
[0038] The term "treating" or "treatment" of a state, disorder or
condition includes:
[0039] (1) preventing or delaying the appearance of clinical
symptoms of the state, disorder or condition developing in a
subject that may be afflicted with or predisposed to the state,
disorder or condition but does not yet experience or display
clinical or subclinical symptoms of the state, disorder or
condition;
[0040] (2) inhibiting the state, disorder or condition, i.e.,
arresting or reducing the development of the disease or at least
one clinical or subclinical symptom thereof; or
[0041] (3) relieving the disease, i.e., causing regression of the
state, disorder or condition or at least one of its clinical or
subclinical symptoms.
[0042] The benefit to a subject to be treated is either
statistically significant or at least perceptible to the subject or
to the physician.
[0043] The term "mean", when preceding a pharmacokinetic value
(e.g., mean peak) represents the arithmetic mean value of the
pharmacokinetic value unless otherwise specified.
[0044] The term "Serum Concentration" or "Serum Concentration
Curve" is the graphic representation of the amount of drug in an
animal's (including humans) plasma at particular points in
time.
[0045] The term "Area Under the Curve" or "Area Under the
Concentration Curve" or "AUC" means the area present beneath the
line of the graphical representation of plasma concentrations
versus time in subject(s). Unless otherwise specified, AUC refers
to the AUC obtained based on baseline adjusted concentrations,
i.e., concentrations obtained after subtracting the individual
baseline from each individual time point (C.sub.t-C.sub.0).
[0046] The term "Cmax" refers to the maximum observed concentration
taken directly from the plasma concentration-time course profile.
Unless otherwise specified, Cmax refers to Cmax obtained based on
baseline adjusted concentrations, i.e., concentrations obtained
after subtracting the individual baseline from each individual time
point (C.sub.t-C.sub.0).
[0047] Delivery Agent Compounds
[0048] In one embodiment of the present invention, the delivery
agent compound has the following structure, or a pharmaceutically
acceptable salt thereof:
##STR00001##
wherein [0049] Ar is phenyl or naphthyl; [0050] Ar is optionally
substituted with one or more of --OH, halogen, C.sub.1-C.sub.4
alkyl, C.sub.1-C.sub.4 alkenyl, C.sub.1-C.sub.4 alkoxy or
C.sub.1-C.sub.4 haloalkoxy; [0051] R.sup.7 is C.sub.4-C.sub.20
alkyl, C.sub.4-C.sub.20 alkenyl, phenyl, naphthyl,
(C.sub.1-C.sub.10 alkyl)phenyl, (C.sub.1-C.sub.10 alkenyl)phenyl,
(C.sub.1-C.sub.10 alkyl)naphthyl, (C.sub.1-C.sub.10
alkenyl)naphthyl, phenyl(C.sub.1-C.sub.10 alkyl),
phenyl(C.sub.1-C.sub.10 alkenyl), naphthyl(C.sub.1-C.sub.10 alkyl),
or naphthyl(C.sub.1-C.sub.10 alkenyl); [0052] R.sup.8 is hydrogen,
C.sub.1 to C.sub.4 alkyl, C.sub.2 to C.sub.4 alkenyl, C.sub.1 to
C.sub.4 alkoxy, or C.sub.1-C.sub.4 haloalkoxy; [0053] R.sup.7 is
optionally substituted with C.sub.1 to C.sub.4 alkyl, C.sub.2 to
C.sub.4 alkenyl, C.sub.1 to C.sub.4 alkoxy, C.sub.1-C.sub.4
haloalkoxy, --OH, --SH, --CO.sub.2R.sub.9, or any combination
thereof; [0054] R.sup.9 is hydrogen, C.sub.1 to C.sub.4 alkyl, or
C.sub.2 to C.sub.4 alkenyl; and [0055] R.sup.7 is optionally
interrupted by oxygen, nitrogen, sulfur or any combination
thereof.
[0056] In one embodiment, the delivery agent compounds are not
substituted with an amino group in the position alpha to the acid
group.
[0057] Suitable delivery agent compounds include, but are not
limited to, N-(8-[2-hydroxybenzoyl]-amino)caprylic acid and salts
thereof, e.g., a sodium salt of
N-(8-[2-hydroxybenzoyl]-amino)caprylic acid, such as a mono- or
di-sodium salt, N-(8-[2-hydroxybenzoyl]-amino) decanoic acid and
pharmaceutically acceptable salts thereof, including its monosodium
salt, 4-[(4-chloro-2-hydroxy-benzoyl)amino]butanoic acid (also
known as 4-[(2-hydroxy-4-chlorobenzoyl)amino]butanoate) and
pharmaceutically acceptable salts thereof, including its sodium
salt (e.g., monosodium salt),
N-(8-[2-hydroxy-5-chlorobenzoyl]-amino)octanoic acid (also known as
8-(N-2-hydroxy-5-chlorobenzoyl)aminocaprylic acid)) and
pharmaceutically acceptable salts thereof, including its monosodium
salt, and 8-(N-2-hydroxy-4-methoxybenzoyl)-aminocaprylic acid and
pharmaceutically acceptable salts thereof, including its monosodium
salt.
[0058] According to one embodiment, R.sup.7 in Formula A is
selected from C.sub.8-C.sub.20 alkyl, C.sub.8-C.sub.20 alkenyl,
phenyl, naphthyl, (C.sub.1-C.sub.10 alkyl)phenyl, (C.sub.1-C.sub.10
alkenyl)phenyl, (C.sub.1-C.sub.10 alkyl) naphthyl,
(C.sub.1-C.sub.10 alkenyl)naphthyl, phenyl(C.sub.1-C.sub.10 alkyl),
phenyl(C.sub.1-C.sub.10 alkenyl), naphthyl(C.sub.1-C.sub.10 alkyl),
and naphthyl(C.sub.1-C.sub.10 alkenyl).
[0059] According to another embodiment, R.sup.7 in Formula A is
selected from C.sub.8-C.sub.20 alkyl, and C.sub.8-C.sub.20
alkenyl.
[0060] In another embodiment of the present invention, the delivery
agent compound has the following structure, or a pharmaceutically
acceptable salt thereof:
##STR00002##
wherein [0061] R.sup.1, R.sup.2, R.sup.3, and R.sup.4 are
independently H, --OH, halogen, C.sub.1-C.sub.4 alkyl,
C.sub.2-C.sub.4 alkenyl, C.sub.1-C.sub.4 alkoxy, --C(O)R.sup.8,
--NO.sub.2, --NR.sup.9R.sup.10, or
--N.sup.+R.sup.9R.sup.10R.sup.11(R.sup.12).sup.-; [0062] R.sup.5 is
H, --OH, --NO.sub.2, halogen, --CF.sub.3, --NR.sup.14R.sup.15,
--N.sup.+R.sup.14R.sup.15R.sup.16(R.sup.13).sup.-, amide,
C.sub.1-C.sub.12 alkoxy, C.sub.1-C.sub.12 alkyl, C.sub.2-C.sub.12
alkenyl, carbamate, carbonate, urea, or --C(O)R.sup.18; [0063]
R.sup.5 is optionally substituted with halogen, --OH, --SH, or
--COOH; [0064] R.sup.5 is optionally interrupted by O, N, S, or
--C(O)--; [0065] R.sup.6 is a C.sub.1-C.sub.12 alkylene,
C.sub.2-C.sub.12 alkenylene, or arylene; [0066] R.sup.6 is
optionally substituted with a C.sub.1-C.sub.4 alkyl,
C.sub.2-C.sub.4 alkenyl, C.sub.1-C.sub.4 alkoxy, --OH, --SH,
halogen, --NH.sub.2, or --CO.sub.2R.sub.8; [0067] R.sup.6 is
optionally interrupted by 0 or N; [0068] R.sup.7 is a bond or
arylene; [0069] R.sup.7 is optionally substituted with --OH,
halogen, --C(O)CH.sub.3, --NR.sup.10R.sup.11, or
--N.sup.+R.sup.10R.sup.11R.sup.12(R.sup.13).sup.-; [0070] each
occurrence of R.sup.8 is independently H, C.sub.1-C.sub.4 alkyl,
C.sub.2-C.sub.4 alkenyl, or --NH.sub.2; [0071] R.sup.9, R.sup.10,
R.sup.11, and R.sup.12 independently H or C.sub.1-C.sub.10 alkyl;
[0072] R.sup.13 is a halide, hydroxide, sulfate, tetrafluoroborate,
or phosphate; [0073] R.sup.14, R.sup.15 and R.sup.16 are
independently H, C.sub.1-C.sub.10 alkyl, C.sub.1-C.sub.10 alkyl
substituted with --COOH, C.sub.2-C.sub.12 alkenyl, C.sub.2-C.sub.12
alkenyl substituted with --COOH, or --C(O)R.sup.17; [0074] R.sup.17
is --OH, C.sub.1-C.sub.10 alkyl, or C.sub.2-C.sub.12 alkenyl; and
[0075] R.sup.18 is H, C.sub.1-C.sub.6 alkyl, --OH,
--NR.sub.14R.sub.15, or
N.sup.+R.sup.14R.sup.15R.sup.16(R.sup.13)--.
[0076] In one particular embodiment, when R.sup.1, R.sup.2,
R.sup.3, R.sup.4, and R.sup.5 are H, and R.sup.7 is a bond then
R.sup.6 is not a C.sub.1-C.sub.6, C.sub.9 or C.sub.10 alkyl.
[0077] In another embodiment, when R.sup.1, R.sup.2, R.sup.3, and
R.sup.4 are H, R.sup.5 is --OH, and R.sup.7 is a bond then R.sup.6
is not a C.sub.1-C.sub.3 alkyl.
[0078] In yet another embodiment, when at least one of R.sup.1,
R.sup.2, R.sup.3, and R.sup.4 is not H, R.sup.5 is --OH, and
R.sup.7 is a bond, then R.sup.6 is not a C.sub.1-C.sub.4 alkyl.
[0079] In yet another embodiment, when R.sup.1, R.sup.2, and
R.sup.3 are H, R.sup.4 is --OCH.sub.3, R.sup.5 is --C(O)CH.sub.3,
and R.sup.6 is a bond then R.sup.7 is not a C.sub.3 alkyl.
[0080] In yet another embodiment, when R.sup.1, R.sup.2, R.sup.4,
and R.sup.5 are H, R.sup.3 is --OH, and R.sup.7 is a bond then
R.sup.6 is not a methyl.
[0081] In yet another embodiment, R.sup.6 of Formula B is a
C.sub.8-C.sub.12 alkylene, C.sub.8-C.sub.12 alkenylene, or
arylene.
[0082] In yet another embodiment of the present invention, the
delivery agent compound has the following structure or a
pharmaceutically acceptable salt thereof:
##STR00003##
wherein
[0083] R.sup.1, R.sup.2, R.sup.3, R.sup.4 and R.sup.5 are
independently H, --CN, --OH, --OCH.sub.3, or halogen, at least one
of R.sup.1, R.sup.2, R.sup.3, R.sup.4 and R.sup.5 being --CN;
and
[0084] R.sup.6 is a C.sub.1-C.sub.12 linear or branched alkylene, a
C.sub.1-C.sub.12 linear or branched alkenylene, a C.sub.1-C.sub.12
linear or branched arylene, an alkyl(arylene) or an
aryl(alkylene).
[0085] According to one embodiment, when R.sup.1 is --CN, R.sup.4
is H or --CN, and R.sup.2, R.sup.3, and R.sup.5 are H, then R.sup.6
is not methylene ((CH.sub.2).sub.1).
[0086] In another embodiment, R.sup.6 of Formula C is a
C.sub.8-C.sub.12 linear or branched alkylene, a C.sub.8-C.sub.12
linear or branched alkenylene, an arylene, an alkyl(arylene) or an
aryl(alkylene).
[0087] In yet another embodiment, R.sup.6 of Formula C is a
C.sub.8-C.sub.12 linear or branched alkylene, a C.sub.8-C.sub.12
linear or branched alkenylene.
[0088] Other suitable delivery agent compounds are disclosed in
U.S. Pat. No. 6,627,228, which is hereby incorporated by
reference.
[0089] The delivery agent compound can also be a a polymeric
delivery agent comprising a polymer conjugated to a modified amino
acid or derivative thereof via a linkage group selected from the
group consisting of --NHC(O)NH--, --C(O)NH--, --NHC(O)--, --OOC--,
--COO--, --NHC(O)O--, --OC(O)NH--, --CH.sub.2NH--, --NHCH.sub.2--,
--CH.sub.2NHC(O)O--, --OC(O)NHCH.sub.2--,
--CH.sub.2NHCOCH.sub.2O--, --OCH.sub.2C(O)NHCH.sub.2--,
--NHC(O)CH.sub.2O--, --OCH.sub.2C(O)NH--, --NH--, --O--, and
carbon-carbon bond. In one embodiment, the polymeric delivery agent
is not a polypeptide or polyamino acid. In another embodiment, the
modified amino acid has the structure of formula A, B, or C. In one
embodiment, the polymeric delivery agent includes a modified amino
acid having the structure:
##STR00004## [0090] which is conjugated via a --COO group to a
polymer having monomers derived from polyethylene glycol.
[0091] In one embodiment, the polymeric delivery agent is a
modified amino acid having the structure of Formula D conjugated
via a --COO group to a polymer having the structure:
--CH.sub.2CH.sub.2O(CH.sub.2CH.sub.2O).sub.xCH.sub.2CH.sub.2O--Y,
[0092] wherein
[0093] x is from 1-14; and
[0094] Y is H or CH.sub.3.
[0095] According to another embodiment, the polymeric delivery
agent is compound having the structure of Formula D conjugated via
a --COO group to a polymer having the structure:
--CH.sub.2CH.sub.2O(CH.sub.2CH.sub.2O).sub.xCH.sub.2CH.sub.2O--Y,
[0096] wherein
[0097] x is 1-5; and
[0098] Y is CH.sub.3 or H.
[0099] For example, the polymeric delivery agent can be
8-(2-hydroxybenzoylamino)-octanoic acid
2-{2-[2-(2-{2-[2-(2-methoxyethoxy)ethoxy]ethoxy}-ethoxy)ethoxy]ethoxy}eth-
yl ester.
[0100] Other suitable delivery agent compounds include compounds of
the formula below and pharmaceutically acceptable salts
thereof:
##STR00005##
[0101] R.sub.1 is --(CH.sub.2).sub.m--R.sub.8, wherein m is 0 or
1;
[0102] R.sub.2-R.sub.6 are independently selected from hydrogen,
hydroxyl, halogen, C.sub.1-C.sub.4 alkyl, C.sub.2-C.sub.4 alkenyl,
C.sub.2-C.sub.4 alkynyl, C.sub.1-C.sub.4 alkoxy, and cyano;
[0103] R.sub.7 is selected from C.sub.1-C.sub.10 alkyl,
C.sub.2-C.sub.10 alkenyl, and C.sub.2-C.sub.10 alkynyl;
[0104] R.sub.8 is selected from cyclopentyl, cyclohexyl and phenyl,
wherein when R.sub.8 is phenyl, m is 1; and
[0105] R.sub.8 is optionally substituted with C.sub.1-C.sub.4
alkyl, C.sub.1-C.sub.4 alkoxy, halogen or hydroxyl, or a
combination thereof.
[0106] Other delivery agent compounds of the present invention
include those of the formula:
##STR00006##
and pharmaceutically acceptable salts thereof, wherein:
[0107] R.sub.1 is a C.sub.1-C.sub.6 alkyl, or C.sub.2-C.sub.6
alkenyl,
[0108] R.sub.2-R.sub.6 are independently selected from hydrogen,
hydroxyl, halogen, C.sub.1-C.sub.4 alkyl, C.sub.2-C.sub.4 alkenyl,
C.sub.2-C.sub.4 alkynyl, C.sub.1-C.sub.4 alkoxy, and cyano, and
[0109] R.sub.7 is selected from C.sub.1-C.sub.10 alkyl,
C.sub.2-C.sub.10 alkenyl, and C.sub.2-C.sub.10 alkynyl.
[0110] Other suitable delivery agent compounds include those of the
formula:
##STR00007##
and pharmaceutically acceptable salts thereof, wherein
[0111] n is 1 to 9, and
[0112] R.sup.1 to R.sup.5 are independently hydrogen, C.sub.1 to
C.sub.6 alkyl, C.sub.1 to C.sub.6 alkoxy, C.sub.2 to C.sub.6
alkenyl, halogen, hydroxyl, --NH--C(O)--CH.sub.3, or
--O--C.sub.6H.sub.5.
[0113] In one embodiment, R.sup.1 to R.sup.5 of Formula G are
independently hydrogen, C.sub.1 to C.sub.4 alkyl, C.sub.1 to
C.sub.4 alkoxy, C.sub.2 to C.sub.4 alkenyl, halogen, or
hydroxyl.
[0114] Other suitable delivery agent compounds include those of the
formula:
##STR00008##
and pharmaceutically acceptable salts thereof, wherein
[0115] R.sup.1 to R.sup.4 are independently hydrogen, C.sub.1 to
C.sub.4 alkyl, C.sub.2 to C.sub.4 alkenyl, halogen, C.sub.1 to
C.sub.4 alkoxy, or hydroxyl.
[0116] Other delivery agent compounds of the present invention
include those of the formula:
##STR00009##
and pharmaceutically acceptable salts thereof, wherein
[0117] one of R.sup.1 to R.sup.5 is --(CH.sub.2).sub.n--COOH where
n is 0-6; and
[0118] the remaining four members of R.sup.1 to R.sup.5 are
independently hydrogen, C.sub.1 to C.sub.4 alkyl, C.sub.2 to
C.sub.4 alkenyl, halogen, C.sub.1 to C.sub.4 alkoxy, or hydroxyl;
and
[0119] R.sub.6-R.sub.10 are independently hydrogen, C.sub.1 to
C.sub.4 alkyl, C.sub.2 to C.sub.4 alkenyl, halogen, C.sub.1 to
C.sub.4 alkoxy, or hydroxyl.
[0120] Other delivery agents of the present invention include
compounds represented by the formula:
##STR00010##
and pharmaceutically acceptable salts thereof, wherein
[0121] n is 1 to 9; and R.sub.1 to R.sub.9 are independently
hydrogen, C.sub.1 to C.sub.4 alkyl, C.sub.2 to C.sub.4 alkenyl,
halogen, C.sub.1 to C.sub.4 alkoxy, or hydroxyl.
[0122] Other suitable delivery agent compounds include those of the
formula:
##STR00011##
and pharmaceutically acceptable salts thereof, wherein
[0123] R.sup.1-R.sup.5 are independently hydrogen, C.sub.1 to
C.sub.4 alkyl, C.sub.2 to C.sub.4 alkenyl, halogen, C.sub.1 to
C.sub.4 alkoxy, hydroxyl, or --O--(CH.sub.2).sub.n--COOH (where n
is 1 to 12);
[0124] at least one of R.sup.1 to R.sup.5 is
--O--(CH.sub.2).sub.n--COOH where n is 1-12; and
[0125] R.sup.6-R.sup.10 are independently hydrogen, C.sub.1 to
C.sub.4 alkyl, C.sub.2 to C.sub.4 alkenyl, halogen, C.sub.1 to
C.sub.4 alkoxy, or hydroxyl.
[0126] Suitable delivery agents are described in International.
Publication Nos. WO 2005/117854 and WO 2005/112633, both of which
were filed May 16, 2005 and their priority documents, U.S.
Provisional Application Nos. 60/576,088, filed Jun. 1, 2004, U.S.
Provisional Application No. 60/576,397, filed Jun. 1, 2004, U.S.
Provisional Application No. 60/576,105, filed Jun. 1, 2004, U.S.
Provisional Application No. 60/571,090, filed May 14, 2004, U.S.
Provisional Application No. 60/571,092, filed May 14, 2004, U.S.
Provisional Application No. 60/571,195, filed May 14, 2004, U.S.
Provisional Application No. 60/571,194, filed May 14, 2004, U.S.
Provisional Application No. 60/571,093, filed May 14, 2004, U.S.
Provisional Application No. 60/571,055, filed May 14, 2004, U.S.
Provisional Application No. 60/571,151, filed May 14, 2004, U.S.
Provisional Application No. 60/571,315, filed May 14, 2004, U.S.
Provisional Application No. 60/571,144, filed May 14, 2004, and
U.S. Provisional Application 60/571,089, filed May 14, 2004, all of
which are hereby incorporated by reference in their entirety.
[0127] Other suitable delivery agents include those having the
following structure and pharmaceutically acceptable salts
thereof:
##STR00012##
wherein
[0128] (a) R.sup.1, R.sup.2, R.sup.3, and R.sup.4 are independently
H, --OH, halogen, C.sub.1-C.sub.4 alkyl, C.sub.1-C.sub.4 alkenyl,
C.sub.1-C.sub.4 alkoxy, --C(O)R.sup.8, --NO.sub.2,
--NR.sup.9R.sup.10, or N.sup.+R.sup.9R.sup.10R.sup.11(Y.sup.-);
[0129] R.sup.8 is hydrogen, --OH, C.sub.1-C.sub.6 alkyl,
C.sub.1-C.sub.4 alkyl substituted with halogen or --OH,
C.sub.2-C.sub.4 alkenyl unsubstituted or substituted with halogen
or --OH, or --NR.sup.14R.sup.15;
[0130] R.sup.9, R.sup.10, and R.sup.H are independently hydrogen,
oxygen, C.sub.1-C.sub.4 alkyl unsubstituted or substituted with
halogen or --OH, C.sub.2-C.sub.4 alkenyl unsubstituted or
substituted with halogen or --OH;
[0131] Y is halide, hydroxide, sulfate, nitrate, phosphate, alkoxy,
perchlorate, tetrafluoroborate, carboxylate, mesylate, fumerate,
malonate, succinate, tartrate, acetate, gluconate, maleate;
[0132] R.sup.5 is H, --OH, --NO.sub.2, halogen, CF.sub.3,
--NR.sup.14R.sup.15, --N.sup.+R.sup.14R.sup.15R.sup.16(Y.sup.-),
amide, C.sub.1-C.sub.12 alkoxy, C.sub.1-C.sub.12 alkyl,
C.sub.2-C.sub.12 alkenyl, carbamate, carbonate, urea, or
--C(O)R.sup.22; R.sup.5 is optionally substituted with halogen,
--OH, --SH, or --COOH; R.sup.5 is optionally interrupted by O, N,
S, or --C(O)--;
[0133] R.sup.14, R.sup.15, and R.sup.16 are independently H or
C.sub.1-C.sub.10 alkyl;
[0134] R.sup.22 is H, C.sub.1-C.sub.6 alkyl, --OH,
--NR.sup.14R.sup.15;
[0135] R.sup.6 is substituted or unsubstituted C.sub.1-C.sub.16
alkylene, C.sub.2-C.sub.16 alkenylene, C.sub.2-C.sub.16 alkynylene,
C.sub.5-C.sub.16 arylene, (C.sub.1-C.sub.16 alkyl) arylene or
aryl(C.sub.1-C.sub.16 alkylene); R.sup.6 is optionally substituted
with C.sub.1-C.sub.7 alkyl or C.sub.1-C.sub.7 cycloalkyl;
[0136] R.sup.7 is NR.sup.18R.sup.19 or
N.sup.+R.sup.18R.sup.19R.sup.20Y.sup.-;
[0137] R.sup.18 and R.sup.19 are independently hydrogen, oxygen,
hydroxy, substituted or unsubstituted C.sub.1-C.sub.16 alkyl,
substituted or unsubstituted C.sub.2-C.sub.16 alkenyl, substituted
or unsubstituted C.sub.2-C.sub.16 alkynyl, substituted or
unsubstituted aryl, substituted or unsubstituted alkylcarbonyl
(e.g. substituted or unsubstituted (C.sub.1-6 alkyl)carbonyl),
substituted or unsubstituted arylcarbonyl, substituted or
unsubstituted alkanesulfinyl (e.g. substituted or unsubstituted
(C.sub.1-6 alkane)sulfinyl), substituted or unsubstituted
arylsulfinyl, substituted or unsubstituted alkanesulfonyl (e.g.
substituted or unsubstituted (C.sub.1-6 alkane)sulfonyl),
substituted or unsubstituted arylsulfonyl, substituted or
unsubstituted alkoxycarbonyl (e.g. substituted or unsubstituted
(C.sub.1-6 alkoxy)carbonyl), or substituted or unsubstituted
aryloxyccarbonyl, or substituted or unsubstituted C.sub.5-C.sub.7
heterocyclic ring (i.e., 5, 6, or 7-membered heterocyclic ring),
wherein the substitutions may be halogen or --OH; and
[0138] R.sup.20 is independently hydrogen, substituted or
unsubstituted C.sub.1-C.sub.16 alkyl, substituted or unsubstituted
C.sub.2-C.sub.16 alkenyl, substituted or unsubstituted
C.sub.2-C.sub.16 alkynyl, substituted or unsubstituted aryl,
substituted or unsubstituted alkylcarbonyl (e.g. substituted or
unsubstituted (C.sub.1-6 alkyl)carbonyl), substituted or
unsubstituted arylcarbonyl, substituted or unsubstituted
alkanesulfinyl (e.g. substituted or unsubstituted (C.sub.1-6
alkane)sulfinyl), substituted or unsubstituted arylsulfinyl,
substituted or unsubstituted alkanesulfonyl (e.g. substituted or
unsubstituted (C.sub.1-6 alkane)sulfonyl), substituted or
unsubstituted arylsulfonyl, substituted or unsubstituted
alkoxycarbonyl (e.g. substituted or unsubstituted (C.sub.1-6
alkoxy)carbonyl), or substituted or unsubstituted aryloxycarbonyl;
or
[0139] (b) R.sup.1-R.sup.16 and R.sup.20 are as defined above;
and
[0140] R.sup.18 and R.sup.19 combine to form a 5, 6, or 7-membered
heterocyclic ring optionally interrupted with an oxo group and
unsubstituted or substituted with C.sub.1-C.sub.6 alkyl,
C.sub.1-C.sub.6 alkoxy, aryl, aryloxy, or carbocyclic ring.
[0141] According to one embodiment, R.sup.7 is morpholino,
morpholinium salt, or diethanolamino.
[0142] According to another embodiment, R.sup.6 is a
C.sub.1-C.sub.16 alkylene and R.sup.7 is morpholino or a
morpholinium salt. Preferably, R.sup.6 is C.sub.4-C.sub.12
alkylene, such as an unsubstituted C.sub.4-C.sub.12 alkylene. More
preferably, R.sup.6 is C.sub.4-C.sub.10, C.sub.4-C.sub.8, or
C.sub.6-C.sub.8 alkylene, such as an unsubstituted
C.sub.4-C.sub.10, C.sub.4-C.sub.8, or C.sub.6-C.sub.8 alkylene.
According to one embodiment, one of R.sup.1-R.sup.5 is hydroxy, for
example, R.sup.1 can be hydroxy.
[0143] According to yet another embodiment, when R.sup.6 is a
C.sub.1-C.sub.10 alkylene, at most one of R.sup.2 and R.sup.4 is
halogen. According to another embodiment, R.sup.6 is a
C.sub.8-C.sub.16, C.sub.9-C.sub.16, C.sub.10-C.sub.16, or
C.sub.11-C.sub.16 alkylene. For instance, R.sup.6 may be a C.sub.8,
C.sub.9, C.sub.10, C.sub.11, or C.sub.12 alkylene (e.g., a normal
C.sub.8-C.sub.12 alkylene). According to yet another embodiment, at
most one of R.sup.1 and R.sup.5 is alkyl.
[0144] According to yet another embodiment, R.sup.1 is hydroxy and
R.sup.2, R.sup.3, R.sup.4, and R.sup.5 are independently hydrogen
or halogen.
[0145] According to yet another embodiment, R.sup.2 is hydroxy and
R.sup.1, R.sup.3, R.sup.4, and R.sup.5 are independently hydrogen
or halogen.
[0146] According to yet another embodiment, R.sup.3 is hydroxy and
R.sup.1, R.sup.2, R.sup.4, and R.sup.5 are independently hydrogen
or halogen.
[0147] In a preferred embodiment, halogen is F, Cl or Br, more
preferably F or Cl, and even more preferably Cl.
[0148] According to yet another embodiment, R.sup.6 is
C.sub.1-C.sub.16 alkylene, (C.sub.1-C.sub.16 alkyl) arylene or
aryl(C.sub.1-C.sub.16 alkylene). More preferably R.sup.6 is
C.sub.1-C.sub.12 alkylene, more preferably C.sub.3-C.sub.10
alkylene, more preferably C.sub.4-C.sub.10 or C.sub.4-C.sub.8
alkylene, and more preferably C.sub.6-C.sub.8 alkylene. More
preferably, R.sup.6 is unsubstituted.
[0149] According to yet another embodiment, R.sup.7 is
--NR.sup.18R.sup.19 and R.sup.18 and R.sup.19 are independently
C.sub.1-C.sub.4 alkyl (e.g., methyl, ethyl, propyl, or butyl)
substituted with --OH. In another embodiment, R.sup.7 is
--NR.sup.18R.sup.19 and R.sup.18 and R.sup.19 combine to form a six
membered heterocyclic ring substituted with an oxo group.
[0150] According to one preferred embodiment, R.sup.1 is hydrogen;
R.sup.2, R.sup.3, and R.sup.4 are independently hydrogen, halogen,
--OH, or --OCH.sub.3; R.sup.5 is hydrogen, --OH, or --C(O)CH.sub.3;
R.sup.6 is C.sub.12 alkylene, and R.sup.7 is NR.sup.18R.sup.19
wherein R.sup.18 and R.sup.19 combine to form a 5, 6 or 7 membered
heterocyclic ring.
[0151] According to another preferred embodiment, one of R.sup.3,
R.sup.4, and R.sup.5 is hydroxy and the others are independently
halogen or hydrogen; R.sup.1 and R.sup.2 are independently halogen
or hydrogen; R.sup.6 is C.sub.1-C.sub.16 alkylene; and R.sup.7 is
NR.sup.18R.sup.19 wherein R.sup.18 and R.sup.19 combine to form a
5, 6, or 7 membered heterocyclic ring. R.sup.6 is preferably
C.sub.6-C.sub.16, C.sub.6-C.sub.10, C.sub.8-C.sub.16,
C.sub.10-C.sub.16, or C.sub.4-C.sub.8 alkylene, such as
unsubstituted C.sub.6-C.sub.16, C.sub.6-C.sub.10, C.sub.8-C.sub.16,
C.sub.10-C.sub.16, or C.sub.4-C.sub.8 alkylene. Preferably,
R.sup.18 and R.sup.19 form a morpholino or imidazole.
[0152] In another preferred embodiment, R.sup.1 is hydrogen;
R.sup.2, R.sup.3, and R.sup.4 are independently hydrogen, halogen,
--OH, or --OCH.sub.3; R.sup.5 is hydrogen, --OH, or --C(O)CH.sub.3;
R.sup.6 is C.sub.1-C.sub.12 alkylene; and R.sup.7 is
N.sup.+R.sup.18R.sup.19R.sup.20 (Y.sup.-) wherein R.sup.18 and
R.sup.19 are hydroxy substituted C.sub.1-C.sub.16 alkyl and
R.sup.20 is hydrogen.
[0153] In another preferred embodiment, R.sup.1 is hydrogen;
R.sup.2, R.sup.3, and R.sup.4 are independently hydrogen, halogen,
--OH, or --OCH.sub.3; R.sup.5 is hydrogen, --OH, or --C(O)CH.sub.3;
R.sup.6 is C.sub.1-C.sub.12 alkylene; and R.sup.7 is
N.sup.+R.sup.18R.sup.19R.sup.20 (Y.sup.-) wherein R.sup.18 and
R.sup.19 are hydroxy substituted C.sub.1-C.sub.16 alkyl and
R.sup.20 is hydrogen.
[0154] In another preferred embodiment, R.sup.1, R.sup.2, R.sup.4,
R.sup.5 are independently halogen or hydrogen; R.sup.3 is --OH, or
--OCH.sub.3; and R.sup.7 is N.sup.+R.sup.18R.sup.19R.sup.20
(Y.sup.-) wherein R.sup.18 and R.sup.19 are hydroxy substituted
C.sub.1-C.sub.16 alkyl and R.sup.20 is hydrogen.
[0155] According to one preferred embodiment, R.sup.1 is hydrogen;
R.sup.2, R.sup.3, and R.sup.4 are independently hydrogen, halogen,
--OH, or --OCH.sub.3; R.sup.5 is hydrogen, --OH, or --C(O)CH.sub.3;
R.sup.6 is C.sub.1-C.sub.6 alkylene or aryl substituted
C.sub.1-C.sub.12 alkyl; and R.sup.7 is --NR.sup.18R.sup.19 wherein
R.sup.18 and R.sup.19 combine to form a 5, 6, or 7 membered
heterocyclic ring or N.sup.+R.sup.18R.sup.19R.sup.20 (Y.sup.-)
wherein R.sup.18 and R.sup.19 are hydroxy substituted
C.sub.1-C.sub.16 alkyl and R.sup.20 is hydrogen.
[0156] In another preferred embodiment, the citrate salt of the
delivery agent is used.
[0157] Other suitable delivery agents include those having the
following structure and pharmaceutically acceptable salts
thereof:
##STR00013##
wherein
[0158] R.sup.1, R.sup.2, R.sup.3, and R.sup.4 are independently H,
--OH, halogen, --OCH.sub.3, --NR.sup.10R.sup.11 or
--N.sup.+R.sup.10R.sup.11R.sup.12(R.sup.13).sup.-;
[0159] R.sup.5 is H, --OH, --NO.sub.2, --NR.sup.14R.sup.15,
--N.sup.+R.sup.14R.sup.15R.sup.16(R.sup.13).sup.-, amide,
C.sub.1-C.sub.12 alkoxy, C.sub.1-C.sub.12 alkyl,
[0160] C.sub.2-C.sub.12 alkenyl, carbamate, carbonate, urea, or
--C(O)R.sup.18;
[0161] R.sup.5 is optionally substituted with --OH, --SH, or
--COOH;
[0162] R.sup.5 is optionally interrupted by O, N, S, or
--C(O)--;
[0163] R.sup.6 is a C.sub.1-C.sub.12 alkylene, C.sub.1-C.sub.12
alkenylene, or arylene;
[0164] R.sup.6 is optionally substituted with a C.sub.1-C.sub.4
alkyl, C.sub.2-C.sub.4 alkenyl, C.sub.1-C.sub.4 alkoxy, --OH, --SH,
halogen, --NH.sub.2, or --CO.sub.2R.sup.9;
[0165] R.sup.6 is optionally interrupted by 0 or N;
[0166] R.sup.7 is a bond or arylene;
[0167] R.sup.7 is optionally substituted with --OH, halogen,
--C(O)CH.sub.3, --NR.sup.10R.sup.11 or
--N.sup.+R.sup.10R.sup.11R.sup.12 (R.sup.13).sup.-;
[0168] R.sup.8 is H or C.sub.1-C.sub.4 alkyl;
[0169] R.sup.9 is H, C.sub.1-C.sub.4 alkyl, or C.sub.2-C.sub.4
alkenyl; R.sup.10, R.sup.11, and R.sup.12 are independently H or
C.sub.1-C.sub.10 alkyl;
[0170] R.sup.13 is a halide, hydroxide, sulfate, tetrafluoroborate,
or phosphate;
[0171] R.sup.14, R.sup.15 and R.sup.16 are independently H,
C.sub.1-C.sub.10 alkyl, C.sub.2-C.sub.12 alkenyl, O, or
--C(O)R.sup.17;
[0172] R.sup.17 is --OH, C.sub.1-C.sub.10 alkyl, or
C.sub.2-C.sub.12 alkenyl; and
[0173] R.sup.18 is --OH, C.sub.1-C.sub.6 alkyl,
--NR.sup.14R.sup.15, --N.sup.+R.sup.14R.sup.15R.sup.16
(R.sup.13).sup.-.
[0174] According to one embodiment, when R.sup.5 is OCH.sub.3 then
R.sup.6 is C.sub.1-C.sub.8 or C.sub.10-C.sub.12 alkyl.
[0175] According to a preferred embodiment, R.sup.5 is not
--OCH.sub.3. More preferably, R.sup.5 is not alkoxy.
[0176] According to another preferred embodiment, R.sup.1, R.sup.2,
R.sup.3, and R.sup.4 are hydrogen, R.sup.5 is --COOH,
--C(O)NH.sub.2, --C(O)CH.sub.3, or --NO.sub.2, R.sup.6 is
--(CH.sub.2).sub.7--, and R.sup.7 is a bond.
[0177] According to yet another preferred embodiment, R.sup.1,
R.sup.2, R.sup.3, and R.sup.4 are hydrogen, R.sup.5 is
--C(O)NH.sub.2, R.sup.6 is --CH.sub.2--, and R.sup.7 is a
para-phenylene.
[0178] According to one embodiment, the delivery agents of formula
(6) have the formula:
##STR00014##
wherein
[0179] R.sup.19 is --NO.sub.2 or --C(O)R.sup.23;
[0180] R.sup.20 is a C.sub.1-C.sub.12 alkylene or C.sub.1-C.sub.12
alkenylene;
[0181] R.sup.21 is a bond or arylene;
[0182] R.sup.22 is H or C.sub.1-C.sub.4 alkyl; and
[0183] R.sup.23 is --OH, C.sub.1-C.sub.6 alkyl, or --NH.sub.2.
[0184] The delivery agent compound can also be any of those
described in U.S. Pat. Nos. 6,699,467, 6,663,898, 6,693,208,
6,693,073, 6,693,898, 6,663,887, 6,646,162, 6,642,411, 6,627,228,
6,623,731, 6,610,329, 6,558,706, 6,525,020, 6,461,643, 6,461,545,
6,440,929, 6,428,780, 6,413,550, 6,399,798, 6,395,774, 6,391,303,
6,384,278, 6,375,983, 6,358,504, 6,346,242, 6,344,213, 6,331,318,
6,313,088, 6,245,359, 6,242,495, 6,221,367, 6,180,140, 6,100,298,
6,100,285, 6,099,856, 6,090,958, 6,084,112, 6,071,510, 6,060,513,
6,051,561, 6,051,258, 6,001,347, 5,990,166, 5,989,539, 5,976,569,
5,972,387, 5,965,121, 5,962,710, 5,958,451, 5,955,503, 5,939,381,
5,935,601, 5,879,681, 5,876,710, 5,866,536, 5,863,944, 5,840,340,
5,824,345, 5,820,881, 5,811,127, 5,804,688, 5,792,451, 5,776,888,
5,773,647, 5,766,633, 5,750,147, 5,714,167, 5,709,861, 5,693,338,
5,667,806, 5,650,386, 5,643,957, 5,629,020, 5,601,846, 5,578,323,
5,541,155, 5,540,939, 5,451,410, 5,447,728, 5,443,841, and
5,401,516; International Publication Nos. WO94/23767, WO95/11690,
WO95/28920, WO95/28838, WO96/10396, WO96/09813, WO96/12473,
WO97/36480, WO 2004/4104018, WO 2004080401, WO 2004062587, WO
2003/057650, WO 2003/057170, WO 2003/045331, WO 2003/045306, WO
2003/026582, WO 2002/100338, WO 2002/070438, WO 2002/069937, WO
02/20466, WO 02/19969, WO 02/16309, WO 02/15959, WO 02/02509, WO
01/92206, WO 01/70219, WO 01/51454, WO 01/44199, WO 01/34114, WO
01/32596, WO 01/32130, WO 00/07979, WO 00/06534, WO 00/06184, WO
00/59863, WO 00/59480, WO 00/50386, WO 00/48589, WO 00/47188, WO
00/46182, WO 00/40203, WO 99/16427, WO 98/50341, WO 98/49135, WO
98/34632, WO 98/25589, WO 98/21951, WO 97/47288, WO 97/31938, WO
97/10197, WO 96/40076, WO 96/40070, WO 96/39835, WO 96/33699, WO
96/30036, WO 96/21464, WO 96/12475, and WO 96/12474; and U.S.
Published Application Nos. 20040110839, 20040106825, 20040068013,
20040062773, 20040022856, 20030235612, 20030232085, 20030225300,
20030198658, 20030133953, 20030078302, 20030072740, 20030045579,
20030012817, 20030008900, 20020155993, 20020127202, 20020120009,
20020119910, 20020102286, 20020065255, 20020052422, 20020040061,
20020028250, 20020013497, 20020001591, 20010039258, and
20010003001. Each of the above listed U.S. patents and U.S. and
International published applications are herein incorporated by
reference.
[0185] Non-limiting examples of delivery agent compounds include
N-(8-[2-hydroxybenzoyl]-amino)caprylic acid,
N-(10-[2-hydroxybenzoyl]-amino)decanoic acid,
8-(2-hydroxy-4-methoxybenzoylamino)octanoic acid,
8-(2,6-dihydroxybenzoylamino)octanoic acid,
8-(2-hydroxy-5-bromobenzoylamino)octanoic acid,
8-(2-hydroxy-5-chlorobenzoylamino)octanoic acid,
8-(2-hydroxy-5-iodobenzoylamino)octanoic acid,
8-(2-hydroxy-5-methylbenzoylamino)octanoic acid,
8-(2-hydroxy-5-fluorobenzoylamino)octanoic acid,
8-(2-hydroxy-5-methoxybenzoylamino)octanoic acid,
8-(3-hydroxyphenoxy)octanoic acid, 8-(4-hydroxyphenoxy)octanoic
acid, 6-(2-cyanophenoxy)hexanoic acid,
8-(2-Hydroxyphenoxyl)octyl-diethanolamine,
8-(4-hydroxyphenoxyl)octanoate, 8-(4-hydroxyphenoxyl)octanoate,
8-(2-hydroxy-4-methoxybenzoylamino)octanoic acid,
8-(2-hydroxy-5-methoxybenzoylamino)-octanoic acid, and salts
thereof. Preferred salts include, but are not limited to,
pharmaceutically acceptable salts thereof such as the monosodium
and disodium salts.
[0186] The delivery agent compounds may be in the form of the
carboxylic acid or pharmaceutically acceptable salts thereof, such
as sodium salts, and hydrates and solvates thereof. The salts may
be mono- or multi-valent salts, such as monosodium salts and
disodium salts. The delivery agent compounds may contain different
counter ions chosen for example due to their effect on modifying
the dissolution profile of the carrier.
[0187] The delivery agent compounds may be prepared by methods
known in the art, such as those discussed in the aforementioned
publications (e.g., International Publication Nos. WO 98/34632, WO
00/07979, WO 01/44199, WO 01/32596, WO 02/20466, and WO 03/045306).
SNAC, SNAD, and the free acid and other salts thereof may be
prepared by methods known in the art, such as those described in
U.S. Pat. Nos. 5,650,386 and 5,866,536.
[0188] Salts of the delivery agent compounds of the present
invention may be prepared by methods known in the art. For example,
sodium salts may be prepared by dissolving the delivery agent
compound in ethanol and adding aqueous sodium hydroxide.
[0189] The delivery agent compound may be purified by
recrystallization or by fractionation on one or more solid
chromatographic supports, alone or linked in tandem. Suitable
recrystallization solvent systems include, but are not limited to,
acetonitrile, methanol, and tetrahydrofuran. Fractionation may be
performed on a suitable chromatographic support such as alumina,
using methanol/n-propanol mixtures as the mobile phase; reverse
phase chromatography using trifluoroacetic acid/acetonitrile
mixtures as the mobile phase; and ion exchange chromatography using
water or an appropriate buffer as the mobile phase. When anion
exchange chromatography is performed, preferably a 0-500 mM sodium
chloride gradient is employed.
Solutions Containing a Delivery Agent Compound and Insulin or an
Analog Thereof
[0190] Some processes of the present invention involve the
introduction of a delivery agent and insulin into an aqueous
solution and obtaining a powder from this solution (e.g. by
lyophillization or rotary evaporation). Generally water is used as
the solvent, although the solvent can also comprise or consist
essentially of other solvents which dissolve the delivery agent
(e.g. sodium 4-CNAB) and insulin. For example, ethanol, methanol,
isopropyl alcohol, tetrahydrofuran, dioxane, butanol, acetone,
2-butanone, methyl tert-butyl ether, n-propanol, methylene
chloride, and other similar low boiling point solvents could be
used in lieu of, or in combination with, water.
[0191] Once obtained, the powder (which contains insulin and a
delivery agent compound) may be further processed (e.g. milled or
granulated with other excipeints) and compressed into tablets or
filled into capsules.
Gelatin Based Pharmaceutical Formulations
[0192] Gelatin is a mixture of purified protein fractions that may
be obtained by partial hydrolysis of animal collagen by an acid or
an alkaline. The process of acid hydrolysis is referred to as Type
A and that by alkaline hydrolysis is referred to as Type B. Gelatin
is a linear polymer that is comprised of amino acids which could
result in a molecular weight ranging from 15,000 to 250,000. As
used herein, the term gelatin includes acid and alkaline
hydrolysates of animal collagen.
[0193] Gelatin may be applied in formulations of the present
invention to serve many functions, such as a coating, a suspending
agent, tablet binder and/or as a viscosity-increasing agent.
Insulin/delivery agent tablets (e.g. insulin/sodium 4-CNAB) may be
formulated at various concentrations of gelatin and at various
ratios of insulin and delivery agent.
[0194] In water, gelatin swells and softens and it can absorb
between 5-10 times its own weight of water. There are several
hydrophilic natural and synthetic polymers may be applied, in
certain embodiments, in place of gelatin. For example, (a) anionic
polymers: alginic acid, dextran sulfate, pectin; (b) cationic acid:
chitosan, polylysine; (c) amphiphatic polymers: carboxylmethyl
chitin, fibrin; (d) neutral polymers such as dextran, agarose,
pullulan.
[0195] As used herein, the term gelatin includes gelatin and
gelatin alternatives disclosed in Remington's Pharmaceutical
Sciences, 16.sup.th ed., Mack Publishing Company, Easton, Pa.
(1980), page 1245 and pages 1576-1582, which is hereby incorporated
by reference in its entirety. The term gelatin also includes
compositions disclosed in U.S. Pat. No. 6,090,915, U.S. Pat. No.
4,043,996, U.S. Pat. No. 4,064,008, U.S. Pat. No. 4,176,117, U.S.
Pat. No. 4,889,920, U.S. Pat. No. 4,374,063, U.S. Pat. No.
5,210,182, U.S. Pat. No. 4,232,425, U.S. Pat. No. 4,402,873, U.S.
Pat. No. 4,427,583, U.S. Pat. No. 5,093,474, U.S. Pat. No.
5,288,408 and U.S. Pat. No. 5,459,241, each of which is hereby
incorporated by reference in their entirety.
[0196] The term gelatin, as used herein also includes gelatin
substitutes and alternatives. Generally, such a gelatin alternative
can be made from easily obtainable (e.g. vegetable) materials
having a homogeneous composition and having all the essential
characteristics of gelatin. In the manufacture of soft gel films
and capsules, the soft gel composition preferably possesses the
properties of good wet and dry film strength, insolubility in cold
water, oil, and alcohol, solubility in hot water, temperature and
pressure sealability, film clarity, film flexibility, edibility,
inertness to drugs or other materials to be encapsulated, and rapid
setting from a hot liquid to form a gel.
[0197] One gelatin alternative is a film-forming composition that
comprises starch material selected from modified starch and waxy
starch; gum; and plasticizer as disclosed in U.S. Pat. No.
6,375,981, which is hereby incorporated by reference. The modified
starch or waxy starch preferably has a dextrose equivalent (DE) of
less than about 1, and more preferably has no measurable DE. This
composition can be, but is not required to be, 100% gelatin-free.
Thus, the composition can be used as a gelatin replacement, or as
an extender in gelatin formulations.
[0198] Another gelatin alternative is wheat fiber gel as disclosed
in U.S. Pat. No. 6,440,480, which is hereby incorporated by
reference. Wheat fiber gel is made by thermal/physical processing
of wheat fiber. A special milling technique is used for treating
wheat material resulting in a product containing a large proportion
of microfine particles. Specific improvements are obtained by
mixing the product with maltodextrin. The product so obtained is
sold under the tradename Vitacel.RTM., by FMC Biopolymer of
Philadelphia, Pa. This product is a dry powder, which readily
disperses in water. Upon stirring of the dispersion the gel forms
through shear forces. It is reported that wheat fiber gel can be
used as a gelatin replacer in yogurt or ice cream. (I. I.
Bollinger, Food Marketing & Techn. October 1995, 4-6).
[0199] Carrageenan is yet another gelatin alternative. Carrageenan
is a natural hydrocolloid, a polysaccharide hydrocolloid, which is
derived from seaweed. It comprises a carbohydrate polymer of
repeating sugar units, which is linear, without significant numbers
of branches or substitutions.
[0200] Methods of Treatment
[0201] The present invention also provides methods for treating a
subject with impaired glucose tolerance or with early or late stage
diabetes comprise orally administering to the mammal a
pharmaceutical formulation of the present invention that includes a
therapeutically effective amount of insulin or an insulin analog
and a delivery agent in an amount effective to facilitate the
absorption of the insulin.
[0202] The pharmaceutical formulations may also include a biguanide
such as metformin, as disclosed in International Application No.
PCT/US05/27499, which is hereby incorporated by reference.
[0203] It is preferred that the administration be on a chronic
basis, e.g., for at least two weeks. In various embodiments, the
administration is preprandially and at bedtime such that, after two
weeks of treatment, the subject achieves improved glucose tolerance
and glycemic control, as well as improved insulin utilization,
insulin sensitivity, insulin secretion capacity and/or HbA.sub.1c
levels, as compared with baseline levels prior to treatment.
[0204] Improved glucose tolerance and better endogenous capacity of
the subject to handle sugar load can also be measured by an AUC of
blood glucose excursion, following a glucose load, that is reduced
by a statistically significant amount as compared with AUC of blood
glucose excursion, following a glucose load, prior to
treatment.
[0205] Improved glycemic control can be demonstrated by: [0206]
decreased fasting blood glucose levels as measured by fasting blood
glucose concentration that is reduced by a statistically
significant amount as compared with baseline fasting blood glucose
concentration prior to treatment. [0207] decreased serum
fructosamine concentrations, as measured by serum fructosamine
assay, that is reduced by a statistically significant amount as
compared with baseline serum fructosamine concentrations prior to
treatment. [0208] improved HbA1c levels after treatment compared
with baseline levels prior to treatment. Preferably, the improved
HbA1c levels are measured by a statistically significant decline in
HbA1c levels. When treating a mammal with impaired glucose
tolerance or with early or late stage diabetes, administration of
the pharmaceutical formulation of the present invention can
preferably be made to a mammal having an HbA.sub.1c level ranging
from normal to elevated prior to treatment.
[0209] Improved insulin utilization and insulin sensitivity of the
subject's body can be measured by a statistically significant
decline in HOMA (Homeostasis Model Assessment). Improved insulin
secretion capacity of the subject's body may also be measured by
Stumvoll first-phase insulin secretion capacity index.
[0210] In preferred embodiments of the invention, by virtue of the
chronic administration of oral dosage forms of the present
invention, the subject achieves improved glucose tolerance and
glycemic control as compared with baseline levels prior to
treatment even without any statistically significant increase in
weight, any statistically significant increase in risk of
hypoglycemia or any statistically significant increase in risk of
hyperinsulinemia in the mammal over the treatment period, and
without the need for monitoring the mammal's blood glucose
concentrations or HbA.sub.1c levels. Further, by virtue of the
chronic administration of oral dosage forms of the present
invention, the subject achieves improved insulin utilization,
insulin sensitivity insulin secretion capacity and HbA.sub.1c
levels as compared with baseline levels prior to treatment.
[0211] It is preferred that the administration of the oral
pharmaceutical formulation is administered 1-4 or more times daily,
preprandially and/or at bedtime. In one embodiment of the
invention, administration of the pharmaceutical formulation takes
place once daily, either at bedtime or preprandially for one meal
during the day time, e.g., for breakfast, lunch or dinner. In
another embodiment, administration of the pharmaceutical
formulation takes place multiple times daily, preferably at bedtime
and preprandially for one meal during the day time, e.g., for
breakfast, lunch or dinner. In a further embodiment, administration
of the pharmaceutical formulation takes place multiple times daily,
preferably at bedtime and preprandially for more than one meal
during the day time. Administration of the pharmaceutical
formulation can also be at or shortly prior to bedtime and
concurrently with or shortly prior to ingestion of each meal, i.e.,
within about 15 minutes or less of ingestion of each meal.
[0212] Preferably, the insulin formulations are administered to
human patients on a chronic basis, e.g., for at least about two
weeks. The dosage form of the present invention can be administered
for at least one day, for one week, for two weeks, for longer
periods, for alternating on-off time periods, or for the life of
the patient.
[0213] The frequency of administration of the oral pharmaceutical
formulation, on a daily basis (i.e., how often during one day-night
period) and on a chronic basis (i.e., for how many days), may
depend upon the patient's position along a "diabetes continuum",
i.e., the extent of the patient's impaired glucose tolerance, the
patient's stage of diabetes and the patient's need for exogenous
glycemic control. This continuum ranges from normal glycemic
control, to simple impaired glucose tolerance and insulin
resistance seen in pre-diabetics or early stage type 2 diabetics,
to failure of insulin production by the pancreas seen in type 1
diabetics and late stage type 2 diabetics. This can also be
measured by the patient's HbA.sub.1c concentration, ranging from
normal to elevated levels (e.g., a HbA1C value of 8.0% or
greater).
[0214] For example, if the subject has a need for fasting glycemic
control, the oral pharmaceutical formulation should preferably be
administered only at or shortly prior to bedtime. If the subject
has some need for post-prandial glycemic control, the oral
pharmaceutical formulation should preferably be administered
preprandially for some meals. If the subject has a need for total
post-prandial glycemic control, the oral pharmaceutical formulation
should preferably be administered preprandially for all meals. If
the subject has a need for comprehensive glycemic control, the oral
pharmaceutical formulation should preferably be administered
preprandially for all meals and at or shortly prior to bedtime.
[0215] Embodiments of the present invention also provide a method
of achieving glucose homeostasis in subjects, comprising orally
administering to a subject a pharmaceutical formulation comprising
a therapeutically effective amount of insulin or an insulin analog
and a delivery agent in an amount effective to facilitate the
absorption of the insulin or insulin analog. It is preferred that
the administration be on a chronic basis, e.g., for at least two
weeks, and be preprandially and at bedtime such that, after two
weeks of treatment, the mammal achieves improved glucose tolerance
and glycemic control as compared with baseline levels prior to
treatment.
EXAMPLES
[0216] The following examples illustrate the invention without
limitation. All parts are given by weight unless otherwise
indicated.
Example 1
Solubilization of Insulin
[0217] The solubility of recombinant human zinc insulin obtained
from Diosynth France (distributed in the U.S. through Diosynth,
Inc.) (hereafter insulin type #1) and Eli Lilly Co. (hereafter
insulin type #2) was determined in aqueous solutions having various
pH values at 37.degree. C. Excess insulin was added to buffer
solutions at pH values of 1, 2, 3, 4, 5, 6, 6.8, and 7.4. Vials
containing the solutions were shaken in a constant temperature
water bath, and visually observed at regular time intervals.
Samples were taken at 2 hours, 4 hours, and 24 hours and analyzed
by a stability-indicating HPLC method. The results are shown below
in Table 1:
TABLE-US-00001 TABLE 1 pH-solubility data of Diosynth and Lilly
insulin at 37.degree. C. Insulin concentration Insulin
concentration Insulin concentration (mg/ml), 2 h (mg/ml), 4 h
(mg/ml), 24 h insulin insulin insulin insulin insulin insulin pH
type # 1 type # 2 type # 1 type # 2 type # 1 type # 2 1.23 >2,
deg* >2, deg >4, deg >4, deg 0.814, deg deg 2.02 >2,
deg >2, deg --, deg --, deg >4, deg >2, deg 3.02 0.989,
deg 0.797, deg 1.092, deg 0.814, deg 0.826, deg 0.731, deg 4.05
0.066 0.068 0.0317 0.040 0.147, deg 0.128, deg 5.07 0.156 0.091
0.155 0.094 0.165, deg 0.124, deg 6.08 0.066 0.049 0.0612 0.044
0.126, deg 0.229, deg 6.82 1.852 0.947 >2 1.294 >4 >2 7.42
>2 >2 >4 >4 >4 >4 deg - products of degradation
present in chromatograms
[0218] Visual observation indicated that insulin type #1 dissolved
faster than insulin type #2. This difference was not observed at pH
1, 2, and 7.4 where dissolution was very rapid for both insulin
types. Solubility at pH 1, 2 and 7.4 was in excess of 4 mg/mL.
Solubility at pH 6.8 was greater than 4 mg/mL for insulin type #1
and 2 mg/mL for insulin type #2, but complete dissolution occurred
only after overnight shaking. The difference in the dissolution
rate of insulin type #2 was most apparent at this pH. Both types of
insulin exhibited some degradation at pH 3, 4, 5, and 6 after 24
hours of shaking. As expected, solubility was lowest around pH 5,
with the solubilities at pH 4, 5, and 6 all being around 0.1
mg/mL.
[0219] The results for pH 3-6.8 are shown in FIG. 1.
Example 2
Solubilization of Insulin in Solutions containing a Delivery
Agent
[0220] The solubility of insulin was investigated in aqueous
solutions containing varying amounts of the delivery agent compound
sodium 4-CNAB. The aqueous solution containing delivery agent was
adjusted to pH 1 by the addition of 1N HCl and excess insulin was
added to the acidified solution. Thereafter, the pH of the solution
was increased by the addition of increments of 1N NaOH. Vials
containing the suspensions were left without shaking or sonicating
for 1-2 hours. The supernatant was analyzed for insulin and sodium
4-CNAB using an HPLC method and the pH was recorded. This process
was repeated with increased amounts of delivery agent added to the
solution until the insulin was completely solubilized. All of these
experiments were performed at room temperature. The results are set
forth below in Table 2.
TABLE-US-00002 TABLE 2 pH of Delivery Delivery Agent, Insulin,
Delivery Agent Agent solution (mg/mL) (mg/mL) None (DI water) 6.3
-- 1.14 4-CNAB 6.4 5 0.5 6.45 10 1 6.26 50 11 6.49 50 44 6.71 100
86 6.45 100 >110 6.83 150 187 6.79 200 >135 SNAC 7.9 150
>200 HPOD 3.9 250 >50
[0221] The results for 4-CNAB and Insulin are also shown in FIG.
2.
Example 4
Solubilization of Insulin Tablet
[0222] A 300 mg pellet of insulin was prepared in a die. The
surface area of the pellet available to the dissolution medium was
0.484 cm.sup.2. The pellet was compressed at 1200-1400 lbs on a
Carver press to form discs. The die was then attached to the shaft
of a dissolution apparatus (USP Dissolution Type II (Paddle) made
by Vankel). The die was rotated at 100 rpm and then immersed in 500
mL of degassed dissolution medium maintained at 37.degree. C.
Dissolution experiments were conducted in water and in aqueous
solutions containing sodium 4-CNAB. Samples of the solutions were
taken over two hours and analyzed by HPLC. FIGS. 3 and 4 show
dissolution profiles of insulin in deionized (DI) water and in 10
mg/mL sodium 4-CNAB solution, respectively.
[0223] The rate of insulin dissolution was significantly greater in
the 10 mg/mL sodium 4-CNAB dissolution media than the deionized
water dissolution media.
[0224] The experiments were repeated in 50 mg/ml sodium 4-CNAB
dissolution media. The insulin levels in these solutions were below
the detection limits of the HPLC technique used.
Example 6
Lyophilized Insulin/4-CNAB Formulations
[0225] Lyophilization as a method of co-drying insulin/4-CNAB
solutions to obtain co-dried insulin/sodium 4-CNAB powder was
investigated. The 3 formulations shown in Table 5 were prepared as
follows.
[0226] Initially, 4-CNAB was used to solubilize the insulin. The
required amounts of insulin and sodium 4-CNAB were weighed out. The
sodium 4-CNAB was added to the required amount of water (about 20
ml of water per gram of sodium 4-CNAB) and stirred (1-5 minutes)
until completely dissolved. The corresponding amount of insulin was
then dispersed in the sodium 4-CNAB solution and left without
stiffing, shaking or sonicating for 0.5-2 hours until solution
became clear. The solution was lyophilized using the cycle in shown
in Table 4.
TABLE-US-00003 TABLE 4 The freeze-drying cycle for Insulin/4-CNAB
solutions. Temperature .degree. C. Time (minutes) Pressure (Torr)
-10 30 -45 30 500 -35 1200 100 -35 240 50 25 1440 25
TABLE-US-00004 TABLE 5 Components Formulation 1 Formulation 2
Formulation 3 Recombinant Human 1.8 mg 1.8 mg 1.8 mg Insulin (50
Units) (Strenght: 27.5 Units/mg) 4-CNAB Monosodium 80 mg 160 mg 240
mg Salt Dibasic Calcium 36 mg 46 mg 79.75 mg Phosphate Magnesium
stearate 1.2 mg 2.2 mg 3.55 mg Total Weight/tablet 119 mg 210 mg
325 mg
[0227] Tablets were prepared by mixing the lyophilized
insulin/4CNAB powder, dibasic calcium phosphate and magnesium
stearate. The powder mixture was compressed into tablets using a
single punch Korsch EK-0 tablet press to prepare an initial
compact. Granules were obtained by crushing the initial compact in
a mortar and passing the granules through a 35 mesh sieve. The
granules were compressed into tablets of a predetermined weight and
stored in a freezer at -20.degree. C.
Example 7
In-vivo Primate Studies
[0228] The formulations of Example 6 were fed to rhesus monkeys.
The monkeys were fasted for at least 12 hrs prior to dosing and up
to 4 hrs after dosing. Water was withheld approximately 1 hr before
dosing and up to 2 hrs after dosing after which it was permitted ad
libitum. The dosing was followed by a 5 ml water flush. Blood
samples (approximately 2 ml each) were collected by venipuncture at
15 minutes before dosing (t=0) and at 5, 10, 15, 20, 30, 45 minutes
and 1, 1.5, 2, 3, 4 hr after dosing.
[0229] Four male primates were administered one tablet of
Formulation 1 of Example 6. The results are shown in FIG. 5
(glucose reduction) and FIG. 6 (serum insulin concentrations). The
averaged results are shown in FIG. 7. After a wash-out period, the
same group of four primates were each administered Formulation 2 of
Example 6. These results are shown in FIGS. 8-10.
[0230] A different group of four male primates were each
administered one tablet of Formulation 3 of Example 6. Glucose
reduction is shown in FIG. 11. Serum insulin levels for 3 of the
four monkeys are shown in FIG. 12.
Example 8
Preparation of Tablets Containing Co-Lyophilized Insulin/4-CNAB
Powder
[0231] Tablets containing co-lypophilized insulin/sodium 4-CNAB
powder and having the formulation shown in Table 6 were prepared as
follows. First, insulin was dissolved in an aqueous solution
containing sodium 4-CNAB and the solution was freeze dried
according to the regimen in Table 4. The insulin/4-CNAB solution
obtained from freeze-drying was then milled with a 35 mesh sieve
and blended with copovidone and magnesium stearate (intragranular
excipents). The composition was then dry granulated by roller
compaction. Copovidone and magnesium stearate was added
extragranularly and the granules were compressed into tablets at a
pressure of about 1000 psi for 5 seconds.
[0232] Based on the process, tablets with the following amounts of
ingredients were prepared:
TABLE-US-00005 TABLE 6 Insulin/4-CNAB (150 units/240 mg)
Co-lyophilized Tablets Weight Weight/Batch Ingredients (mg/tablet)
(g) Insulin/4-CNAB (Co-lyophilized 5.45/240 270.0 containing 150
Units of insulin) Copovidone; NF/EP (intragranular) 3.60 3.96
Magnesium Stearate; NF/EP(intragranular) 0.90 0.99 Copovidone;
NF/EP(extragranular) 3.60 3.96 Dibasic Calcium Phosphate Anhydrous,
103.75 114.13 USP/EP Magnesium Stearate 2.70 2.97 Theoretical
Tablet Weight 360 mg
Example 9
Preparation of Tablets Containing Co-Lyophilized Insulin/4-CNAB
Powder
[0233] Tablets containing co-lyophilized insulin/4-CNAB and having
the formulation shown in Table 7 were prepared by the procedure set
forth in Example 8.
TABLE-US-00006 TABLE 7 Insulin/4-CNAB (150 units/400 mg)
Co-lyophilized Tablets Weight Weight/Batch Ingredients (mg/tablet)
(g) *Insulin/4-CNAB (Co-lyophilized) 5.45/400 430.588 (Containing
150 units of Insulin) Copovidone; NF/EP (intragranular) 5.80 6.160
Magnesium Stearate; NF/EP(intragranular) 1.40 1.487 Copovidone;
NF/EP(extragranular) 5.75 6.106 Dibasic Calcium Phosphate
Anhydrous, 157.20 166.946 USP/EP Magnesium Stearate 4.40 4.673
Theoretical Tablet Weight 580 mg
Example 10
Preparation of Tablets Containing Co-Dried Insulin/4-CNAB Powder
Obtained from Rotary Evaporation
[0234] Sodium 4-CNAB (935.7 g, 0.4 wt % water by Karl Fisher
titration) and purified water (4850 mL) were charged to a 20 L,
borosilicate, rotary evaporator flask. The flask was attached to a
rotary evaporator and rotated at about 60 rpm until the solids
dissolved. The rotation was stopped. The flask was removed from the
rotary evaporator and insulin (64.1 g, 27.4 U/mg) was added. The
flask was re-attached to the rotary evaporator and the insulin was
allowed to dissolve without agitation. The water was removed
rapidly with the rotary evaporator bath set at about 45 C and the
internal pressure set at about 5 mm Hg. The walls of the rotary
evaporator flask became coated with solid co-dried insulin/4CNAB as
the water was removed. These solids were scraped from the walls of
the flask and dried in a vacuum oven set at full vacuum and
50.degree. C. until the water content was less than 10 wt % by Karl
Fisher analysis. In most cases the vacuum drying time could be
reduced by breaking up the larger lumps of co-dried material about
midway through the vacuum oven drying cycle. The dried material was
then hammer milled so that it would pass through a 35 mesh screen.
The milled, powdery material was placed in a suitable container and
stored in a freezer at -20.degree. C. or lower until used. The
recovery of co-dried material was about 95%.
[0235] The co-dried insulin/4-CNAB prepared as described above, was
mixed with povidone and magnesium stearate in the amounts shown in
Table 8 below and compressed into tablets tablets at a pressure of
about 1000 psi for 5 seconds.
TABLE-US-00007 TABLE 8 Ingredient Amount 4-CNAB Sodium Salt 80 mg
Recombinant Human Insulin 150 Units (27.4 U/mg) Povidone, USP
(Kollidon 90F) 1 mg Magnesium stearate, NF 1 mg
Example 11
Preparation of Capsules Containing Co-Dried Insulin/4-CNAB Powder
Obtained From Rotary Evaporation
[0236] The process of example 10 was repeated to prepare the
co-dried insulin/4-CNAB powder. 80 mg of sodium 4-CNAB/150 Units of
insulin was placed, without excipients, into size 2 hard gelatin
opaque white capsules.
Example 12
Preparation of Tablets Containing Co-Dried Insulin/4-CNAB Powder
Obtained from Rotary Evaporation
[0237] The process of example 10 was repeated to prepare the
co-dried insulin/4-CNAB powder, except that 966.7 g of 4-CNAB, 5000
mL of water and 33.23 g of insulin were charged to the rotary
evaporator flask.
[0238] Tablets having the formulation shown in Table 9 were
prepared by the procedure set forth in Example 10:
TABLE-US-00008 TABLE 9 Ingredient Amount 4-CNAB Sodium Salt 80 mg
Recombinant Human Insulin 75 Units (27.4 U/mg) Povidone, USP
(Kollidon 90F) 1 mg Magnesium stearate, NF 1 mg
Example 13
Preparation of Capsules Containing Co-Dried Insulin/4-CNAB Powder
Obtained from Rotary Evaporation
[0239] The process of example 10 was repeated to prepare the
co-dried insulin/4-CNAB powder, except that 966.7 g of 4-CNAB, 5000
mL of water and 33.23 g of insulin were charged to the rotary
evaporator flask.
[0240] 80 mg of sodium 4-CNAB/75 Units of insulin was placed,
without excipients, into size 2 hard gelatin opaque white
capsules.
Example 14
Preparation of Tablets Containing Co-Dried Insulin/4-CNAB Powder
Obtained from Rotary Evaporation
[0241] The process of Example 10 was repeated to prepare tablets,
except that povidone and magnesium stearate were not included in
the formulation.
Example 15
Preparation of Tablets Containing Co-Dried Insulin/4-CNAB Powder
Obtained from Rotary Evaporation
[0242] The process of Example 10 was repeated to prepare the
co-dried insulin/4-CNAB powder, except that 241.7 g of 4-CNAB, 1260
mL of water and 8.3 g of insulin were charged to the rotary
evaporator flask.
[0243] Tablets having the formulation shown in Table 10 were
prepared by the procedure set forth in Example 10.
TABLE-US-00009 TABLE 10 Ingredient Amount 4-CNAB Sodium Salt 160 mg
Recombinant Human Insulin 150 Units (27.4 U/mg) Povidone, USP
(Kollidon 90F) 0 mg Magnesium stearate, NF 0 mg
Example 16
Preparation of Tablets Containing Co-Dried Insulin/4-CNAB Powder
Obtained from Rotary Evaporation
[0244] The process of example 10 was repeated to prepare the
co-dried insulin/4-CNAB powder, except that 245.8 g of 4-CNAB, 1280
mL of water and 4.3 g of insulin were charged to the rotary
evaporator flask.
[0245] Tablets having the formulation shown in Table 11 were
prepared by the procedure set forth in Example 10.
TABLE-US-00010 TABLE 11 Ingredient Amount 4-CNAB Sodium Salt 320 mg
Recombinant Human Insulin 150 Units (27.4 U/mg) Povidone, USP
(Kollidon 90F) 0 mg Magnesium stearate, NF 0 mg
Example 17
Preparation of Tablets Containing Co-Dried Insulin/4-CNAB Powder
Obtained from Rotary Evaporation
[0246] The process of example 10 was repeated to prepare the
co-dried insulin/4-CNAB powder, except that 245.4 g of 4-CNAB, 1280
mL of water and 5.6 g of insulin were charged to the rotary
evaporator flask.
[0247] Tablets having the formulation shown in Table 12 were
prepared by the procedure set forth in Example 10.
TABLE-US-00011 TABLE 12 Ingredient Amount 4-CNAB Sodium Salt 160 mg
Recombinant Human Insulin 100 Units (27.4 U/mg) Povidone, USP
(Kollidon 90F) 0 mg Magnesium stearate, NF 0 mg
Example 18
Preparation of Tablets Containing Co-Dried Insulin/4-CNAB Powder
Obtained from Rotary Evaporation
[0248] The process of example 10 was repeated to prepare the
co-dried insulin/4-CNAB powder, except that 247.2 g of 4-CNAB, 1300
mL of water and 2.8 g of insulin were charged to the rotary
evaporator flask.
[0249] Tablets having the formulation shown in Table 13 were
prepared by the procedure set forth in Example 10.
TABLE-US-00012 TABLE 13 Ingredient Amount 4-CNAB Sodium Salt 320 mg
Recombinant Human Insulin 100 Units (27.4 U/mg) Povidone, USP
(Kollidon 90F) 0 mg Magnesium stearate, NF 0 mg
Example 19
Preparation of Tablets Containing Co-Dried Insulin/4-CNAB Powder
Obtained from Rotary Evaporation
[0250] The process of example 10 was repeated to prepare the
co-dried insulin/4-CNAB powder, except that 247.2 g of 4-CNAB, 1300
mL of water and 2.8 g of insulin were charged to the rotary
evaporator flask.
[0251] Tablets having the formulation shown in Table 14 were
prepared by the procedure set forth in Example 10.:
TABLE-US-00013 TABLE 14 Ingredient Amount 4-CNAB Sodium Salt 320 mg
Recombinant Human Insulin 100 Units (27.4 U/mg) Povidone, USP
(Kollidon 90F) 1 mg Magnesium stearate, NF 1 mg
Example 20
Preparation of Tablets Containing Co-Dried Insulin/4-CNAB Powder
Obtained from Rotary Evaporation
[0252] The process of example 10 was repeated to prepare the
co-dried insulin/4-CNAB powder, except that 245.8 g of 4-CNAB, 1280
mL of water and 4.3 g of insulin were charged to the rotary
evaporator flask.
[0253] Tablets having the formulation shown in Table 15 were
prepared by the procedure set forth in Example 10.
TABLE-US-00014 TABLE 15 Ingredient Amount 4-CNAB Sodium Salt 320 mg
Recombinant Human Insulin 150 Units (27.4 U/mg) Povidone, USP
(Kollidon 90F) 1 mg Magnesium stearate, NF 1 mg
Example 21
Preparation of Tablets Based on Granulated Gelatin-Based
Formulation
[0254] Tablets having the formulation shown in Table 16 were
prepared as follows. Gelatin was milled using a Kitchen-Aid.TM.
coffee grinder and later screened through a sieve of size #35.
Afterwards, the required amount of insulin was weighed and
dispersed into purified water. The amount of purified water used
was about 15% of the theoretical batch weight. For a theoretical
batch size of 1200 tablets, 50 g of Insulin is dispersed in 46.7 g
of purified water.
[0255] Sodium 4-CNAB and the milled gelatin were transferred into
an appropriate sized high shear granulator and mixed for about 2
minutes. The sodium 4-CNAB and gelatin mixture was granulated first
with the aqueous dispersion of insulin and later with purified
water using a pump device. The resultant wet granules were evenly
dispersed in oven trays and dried in a vacuum oven
(Temperature=50.degree. C.; Vacuum=5 mm Hg) for at least 8 hours.
The dried granules were characterized based on moisture (0.5%) and
insulin content and insulin content uniformity. The granules were
milled and screened through a sieve of size 0.02 inches. Prior to
tablet compression, dibasic calcium phosphate and magnesium
stearate were blended with dry granules. Tablets were compressed
using an EKO single punch station press.
[0256] Based on the above procedure, tablets containing the
following amounts of ingredients were prepared.
TABLE-US-00015 TABLE 16 Weight Ingredients (mg/dose) Recombinant
Human Insulin 5.45 4-CNAB Monosodium salt 240 Gelatin (Type A) 12
Dibasic Calcium Phosphate (extragranular) 113.80 Magnesium Stearate
(extragranular) 3.75 Total Weight (mg/tablet) 375
[0257] The tablets prepared by this process had an average weight
of about 373.5 mg, a thickness of 5 mm and an average hardness of
about 10.3 kP.
Example 22
Preparation of Granulated Tablets (Wet Granulation)
[0258] Tablets having the formulation shown in Tables 17 and 18
were prepared as follows.
TABLE-US-00016 TABLE 17 Drug Product Components: Insulin/4- CNAB
(150 Units/80 mg) Tablets Component Function 4-CNAB Sodium Salt
Delivery Agent Recombinant Human Insulin Drug (Active Agent)
Povidone, USP (Kollidon 90F) Binder Dibasic Calcium Phosphate,
Anhydrous, USP Binder Magnesium stearate, NF Lubricant Purified
Water, USP Granulating fluid
[0259] Sodium 4-CNAB was milled using a Quadro Comil equipped with
a 35 mesh screen. Insulin and the milled sodium 4-CNAB were blended
together, and transferred to a Key Instruments KG 5 high shear
granulator equipped with a 5 Liter bowl. The material was
granulated with povidone. Once the addition of the povidone was
completed, the container was rinsed with small portions of purified
water and added to the granulation until the desired granulation
was achieved.
[0260] The granulation was transferred to clean stainless steel
trays and dried in a vacuum oven at 50.degree. C. until the
moisture content was less than 5.0% w/w and then milled through a
35 mesh screen and further dried until the moisture content less
than 1.5% w/w. The granulation was assayed for insulin and sodium
4-CNAB using a validated HPLC method. The insulin assay of the
granulation was used for calculating the required quantity of
Emcompress for the batch. The required amount of Emcompress was
added to the granulation and blending was performed in a V-blender
for 15 minutes. Samples were collected for bend uniformity testing.
After acceptable blend uniformity data was obtained the required
amount of magnesium stearate was added and blending was performed
for 3 minutes. The resulting blend was compressed into tablets
using a Korsch EKO single station tablet press. The target tablet
weight was 125 mg with a range of 119-131 mg, acceptable tablet
hardness range was 5-11 kP with a target tablet hardness of 7.0 kP.
The tablets exhibited an average thickness of 7.8 mm. The
Insulin/4-CNAB (150 Units/80 mg) tablets were packaged in a
container closure system consisting of a 60 cc HDPE Round, White
bottle with 33 mm Child Resistant Cap 1 with Safe-Gard.RTM. 75 m
Induction Innerseal and Cotton Coil 12 gm/yd.
[0261] Based on this procedure, the following amounts of
ingredients were used to prepare the tablets:
TABLE-US-00017 TABLE 18 Drug Product Composition: Insulin/4-CNAB
(150 Units/80 mg) Tablets (Batch Size 6,200 Tablets) Component
mg/Tablet Batch Formula (g) 4-CNAB Sodium Salt 76-84 496.0
Recombinant Human Insulin 5.32-5.88 35.1 (142.5-157.5 Units)
Povidone USP (Kollidon 90F) 0.38-0.42 2.2 Dibasic Calcium
Phosphate, 35.9-39.7 195.2 Anhydrous, USP Magnesium Stearate, NF
1.14-1.26 6.7 Impalpable Powder Total Weight (mg) 118.8-131.3
735.2
Example 23
Human Clinical Study of Orally Administered Insulin
[0262] Six healthy male subjects between the ages of 18 and 40 were
orally administered one tablet or capsule, depending on the
treatment period indicated below after an 8-hour fast the previous
night. Glucose and insulin values were obtained from blood samples
fifteen minutes prior to dosing (t=0) and 5, 10, 15, 20, 25, 30,
35, 40, 45, 50, 60, 90 and 120 minutes after dosing. After
receiving the first tablet or capsule, the subject underwent at
least a 72 hour washout period before receiving the next tablet or
capsule.
[0263] The administration regimen was as follows:
TABLE-US-00018 TABLE 19 Treatment Period Summary of Dosage
Preparation Details A Wet granulation tablet Example 22 B Rotary
evaporation tablet Example 10 C Rotary evaporation capsule Example
11 D Rotary evaporation tablet Example 12 E Rotary evaporation
capsule Example 13
[0264] Results for the treatment regiment are set forth below:
TABLE-US-00019 TABLE 20 Insulin Mean PK Results obtained from
Individual Baseline-adjusted Insulin Concentrations Cmax Tmax
AUClast New/Reference New/Reference Treatment (.mu.U/mL) (min)
(.mu.U/mL * min) Cmax ratio AUC ratio A N 6 6 6 N/A N/A Mean 15.250
19.167 346.250 SD 10.064 2.041 227.404 Min 4.50 15.00 40.00 Max
31.00 20.00 700.00 CV % 66.0 10.6 65.7 B N 6 6 6 1.8 1.2 Mean
27.500 19.167 422.708 SD 9.925 10.685 256.758 Min 14.50 10.00
133.75 Max 41.50 40.00 860.00 CV % 36.1 55.7 60.7 C N 5 5 5 Mean
8.800 35.000 260.000 SD 7.497 33.727 313.358 Min 0.50 10.00 11.25
Max 21.00 90.00 805.00 CV % 85.2 96.4 120.5 C N 5 5 5 0.2 0.4
(without Mean 5.200 41.000 128.000 outlier- SD 3.347 46.287 73.912
Subject 4, Min 0.50 10.00 11.25 90 min Max 8.50 120.00 200.00
timepoint) CV % 64.4 112.9 57.7 D N 6 6 6 0.3 0.3 Mean 9.333 10.000
110.000 SD 4.215 3.162 59.713 Min 1.00 5.00 7.50 Max 12.50 15.00
172.50 CV % 45.2 31.6 54.3 E N 5 4 5 0.4 0.4 Mean 5.600 38.750
140.500 SD 6.004 37.053 123.392 Min 0.00 5.00 0.00 Max 15.50 90.00
273.75 CV % 107.2 95.6 87.8
TABLE-US-00020 TABLE 21 Insulin Mean PK Results obtained from
Individual Insulin Concentrations Cmax Tmax AUClast New/Reference
New/Reference Treatment (.mu.U/mL) (min) (.mu.U/mL * min) Cmax
ratio AUC ratio A N 6 6 6 N/A N/A Mean 30.333 19.167 1882.917 SD
11.343 2.041 435.067 Min 16.00 15.00 1107.50 Max 44.00 20.00
2222.50 CV % 37.4 10.6 23.1 B N 6 6 6 1.6 1.2 Mean 47.167 19.167
2286.250 SD 14.148 10.685 1117.921 Min 21.00 10.00 747.50 Max 59.00
40.00 4222.50 CV % 30.0 55.7 48.9 C(*) N 6 6 6 0.6 0.8 Mean 18.667
27.500 1562.083 SD 4.179 45.689 247.454 Min 12.00 0.00 1127.50 Max
24.00 120.00 1777.50 CV % 22.4 166.1 15.8 D N 6 6 6 0.8 0.9 Mean
24.500 10.000 1640.417 SD 6.863 3.162 522.110 Min 18.00 5.00 867.50
Max 36.00 15.00 2500.00 CV % 28.0 31.6 31.8 E N 6 6 6 0.9 0.9 Mean
26.167 26.667 1744.167 SD 15.052 34.303 414.203 Min 16.00 0.00
1102.50 Max 54.00 90.00 2222.50 CV % 57.5 128.6 23.7 (*)Insulin
concentration measured at 90 min in Subject 4 was considered an
outlier hence not included in the PK analysis.
[0265] Model-independent pharmacokinetic metrics were calculated
using WinNonlin (v 4.5, Scientific Consulting, Inc.) from
individual plasma concentration data. This program analyzes data
using the standard methods described by Gibaldi and Perrier. The
area under the plasma concentration-time curve (AUC) was estimated
by the linear trapezoidal rule.
[0266] "Baseline` is defined as average plasma insulin
concentrations as assessed prior to study drug administration (i.e.
-15 and 0 minute samples averaged). Adjusted concentrations were
obtained by subtracting the individual baseline from each
individual time point (Ct-Co). Negative values were not included in
the data analysis.
[0267] Results for the adjusted-mean insulin profiles are shown in
FIGS. 13 (without outlier) and 14 (with outlier).
Example 24
Human Clinical Study of Orally Administered Insulin
[0268] The protocol used in Example 23 was used for the following
treatment regimens:
TABLE-US-00021 TABLE 22 Treatment Period Summary of Dosage
Preparation Details A Rotary Evaporation Tablets Example 10 B
Rotary Evaporation Tablets Example 14 C Rotary Evaporation Tablets
Example 15 D Rotary Evaporation Tablets Example 16 E Rotary
Evaporation Tablets Example 17 F Rotary Evaporation Tablets Example
18
[0269] Results for the treatment regiment are set forth below:
TABLE-US-00022 TABLE 23 Insulin Mean PK Results obtained from
Individual Baseline-adjusted Insulin Concentrations Cmax Tmax
AUClast Treatment (.mu.U/mL) (min) (.mu.U/mL*min) A N 6 6 6 Mean
30.633 17.500 795.615 SD 20.977 5.244 542.094 Min 10.86 10.00
331.76 Max 66.62 25.00 1742.30 CV % 68.5 30.0 68.1 B N 6 6 6 Mean
22.441 13.333 312.535 SD 16.489 2.582 214.398 Min 8.00 10.00 118.83
Max 52.00 15.00 649.80 CV % 73.5 19.4 68.6 C N 6 6 6 Mean 28.546
21.667 741.708 SD 23.848 14.024 737.074 Min 6.05 5.00 100.93 Max
69.20 45.00 1787.75 CV % 83.5 64.7 99.4 D N 6 6 6 Mean 29.421
16.667 575.435 SD 17.155 4.082 384.295 Min 13.56 10.00 176.14 Max
53.08 20.00 1210.13 CV % 58.3 24.5 66.8 E N 6 6 6 Mean 18.556
15.833 303.702 SD 14.849 2.041 304.338 Min 1.49 15.00 13.35 Max
39.25 20.00 799.38 CV % 80.0 12.9 100.2 F N 6 6 6 Mean 20.971
19.167 536.952 SD 16.710 9.704 528.919 Min 2.14 5.00 15.06 Max
44.11 30.00 1312.61 CV % 79.7 50.6 98.5
TABLE-US-00023 TABLE 24 Insulin Mean PK Results obtained from
Individual Insulin Concentrations Cmax Tmax AUClast Treatment
(.mu.U/mL) (min) (.mu.U/mL*min) A N 6 6 6 Mean 41.945 17.500
1905.513 SD 23.681 5.244 880.650 Min 14.94 10.00 794.20 Max 81.36
25.00 3251.53 CV % 56.5 30.0 46.2 B N 6 6 6 Mean 35.740 13.333
1590.567 SD 14.956 2.582 246.447 Min 20.30 10.00 1199.00 Max 63.19
15.00 1961.45 CV % 41.8 19.4 15.5 C N 6 6 6 Mean 40.807 21.667
1955.696 SD 24.954 14.024 896.595 Min 15.60 5.00 1032.25 Max 84.17
45.00 3295.53 CV % 61.2 64.7 45.8 D N 6 6 6 Mean 42.480 16.667
1919.300 SD 18.411 4.082 493.982 Min 26.08 10.00 1363.75 Max 68.64
20.00 2586.25 CV % 43.3 24.5 25.7 E N 6 6 6 Mean 31.770 15.833
1563.100 SD 13.306 2.041 334.769 Min 13.95 15.00 1030.88 Max 49.90
20.00 1984.25 CV % 41.9 12.9 21.4 F N 6 6 6 Mean 33.737 19.167
1861.629 SD 16.049 9.704 654.066 Min 17.02 5.00 1271.05 Max 53.39
30.00 2969.95 CV % 47.6 50.6 35.1
[0270] Results based on the adjusted mean insulin profiles are set
forth in FIG. 15
Example 25
Human Clinical Study of Orally Administered Insulin
[0271] The protocol used in Example 23 was used for the following
treatment regimens:
TABLE-US-00024 TABLE 25 Treatment Period Summary of Dosage
Preparation Details A Gelatin-Based Tablet Example 21 B
Co-lyophilized Tablets Example 8 C Rotary Evaporation Tablets
Example 19 D Rotary Evaporation Tablets Example 20 E Wet
granulation Tablets Example 22 F Co-lyophilized Tablets Example
9
[0272] Results for the treatment regiment are set forth below:
TABLE-US-00025 TABLE 26 Insulin Mean PK Results obtained from
Individual Baseline-adjusted Insulin Concentrations Cmax Tmax
AUClast New/Reference New/Reference Treatment (.mu.U/mL) (min)
(.mu.U/mL * min) Cmax ratio AUC ratio A N 6 6 6 2.6 3.6 Mean 30.633
17.500 764.871 SD 20.977 5.244 508.089 Min 10.86 10.00 331.76 Max
66.62 25.00 1629.01 CV % 68.5 30.0 66.4 B N 6 6 6 0.9 1.3 Mean
10.531 32.500 286.569 SD 12.685 43.215 284.348 Min 1.33 10.00 12.68
Max 33.61 120.00 731.20 CV % 120.5 133.0 99.2 C N 6 6 6 1.0 1.2
Mean 12.203 15.000 254.679 SD 11.976 3.162 256.555 Min 4.89 10.00
50.65 Max 36.15 20.00 755.21 CV % 98.1 21.1 100.7 D N 6 6 6 1.6 1.9
Mean 19.013 13.333 415.721 SD 24.735 6.055 601.162 Min 2.12 5.00
9.80 Max 67.82 20.00 1576.64 CV % 130.1 45.4 144.6 E N 6 6 6 N/A
N/A Mean 11.693 17.500 213.752 SD 10.033 8.216 180.716 Min 0.69
10.00 1.73 Max 26.70 30.00 415.56 CV % 85.8 46.9 84.5 F N 6 6 6 0.8
0.7 Mean 9.092 15.000 142.221 SD 5.539 8.367 77.332 Min 1.83 10.00
4.56 Max 18.54 30.00 232.71 CV % 60.9 55.8 54.4
TABLE-US-00026 TABLE 27 Insulin Mean PK Results obtained from
Individual Insulin Concentrations Cmax Tmax AUClast New/Reference
New/Reference Treatment (.mu.U/mL) (min) (.mu.U/mL * min) Cmax
ratio AUC ratio A N 6 6 6 1.4 1.6 Mean 32.098 36.667 1966.079 SD
13.956 40.947 963.466 Min 9.85 15.00 800.78 Max 47.49 120.00
3345.18 CV % 43.5 111.7 49.0 B N 6 6 6 1.0 1.1 Mean 21.730 32.500
1379.888 SD 10.133 43.215 281.651 Min 14.06 10.00 1151.90 Max 38.54
120.00 1922.78 CV % 46.6 133.0 20.4 C N 6 6 6 1.1 1.2 Mean 23.873
15.000 1437.488 SD 12.681 3.162 401.442 Min 12.92 10.00 927.30 Max
48.83 20.00 2129.28 CV % 53.1 21.1 27.9 D N 6 6 6 1.3 1.2 Mean
29.630 13.333 1492.971 SD 25.993 6.055 895.031 Min 7.20 5.00 398.93
Max 78.99 20.00 2857.18 CV % 87.7 45.4 59.9 E N 6 6 6 N/A N/A Mean
22.507 17.500 1239.967 SD 8.105 8.216 244.282 Min 14.67 10.00
878.80 Max 32.70 30.00 1507.50 CV % 36.0 46.9 19.7 F N 6 6 6 0.8
0.8 Mean 18.093 15.000 1030.767 SD 6.660 8.367 287.163 Min 11.84
10.00 648.45 Max 29.57 30.00 1360.88 CV % 36.8 55.8 27.9
[0273] Results based on the adjusted mean insulin profiles are set
forth in FIG. 16
Example 26
Human Clinical Study of Orally Administered Insulin
[0274] 145 humans patients having type 2 diabetes who were failing
metformin monotherapy were enrolled in a 90 day randomized
double-blind placebo-controlled study. The patients remained on
their metformin regimen, which varied from 125 mg-3000 mg per day
individualized for each patient. 141 patients completed the
study.
[0275] The patients were separated into 4 treatment groups as
follows:
TABLE-US-00027 Group Insulin Dose Dosing Regimen 1 (n = 35) Tablet
of Example 22 2 insulin tablets, 4 times daily (150 IU of insulin
each) 2 (n = 35) Tablet of Example 22 2 insulin tablets, 2 times
daily (150 IU of insulin) 2 placebo tablets, 2 times daily 3 (n =
36) Tablet of Example 22 1 insulin tablet + 1 placebo (150 IU of
insulin) tablet, 4 times daily 4 (n = 35) Placebo 2 placebo
tablets, 4 times daily
[0276] Hemoglobin A1c counts were obtained about three weeks prior
to beginning the study (screening) and just prior to the first
administration of the oral insulin (t=0) or (baseline).
[0277] Changes in HbA1c levels at the conclusion of the 90 day
study for the four groups (relative to baseline) are shown in FIG.
17.
[0278] FIGS. 18 and 19 shows the difference between Hemoglobin A1c
level at screening and at baseline for each of the subjects.
[0279] FIG. 20 is a bar graph of the changes in HbA1c level at 90
days compared to baseline for populations in groups 1 and 4 having
a difference in HbA1c levels between screening and baseline of, for
Group 1, 0-1.1, 0-0.5 and 0-0.3 and, for Group 4, 0-1, 0-0.5 and
0-0.3.
[0280] FIG. 21 is a bar graph of the change in HbA1c values after
90 days (compared to baseline) for patients in groups 1 and 4
having particular baseline HbA1c values and a difference in the
HbA1c values from screening to baseline of 0-0.3. The first bar
represents absolute changes in HbA1c levels (t=0 vs. t=90) for
those subjects in Group I having baseline HbA1c levels ranging from
7 to 8.9 and a 0-0.3 variation of HbA1c levels between screening
and at baseline. The second bar represents absolute changes in
HbA1c levels for a subpopulation of Group 1 having baseline HbA1c
levels of 7.5 to 8.9 and a 0-0.3 variation of HbA1c levels between
screening and at baseline. The third bar represents a still
narrower subpopulation--those patients of Group I having baseline
HbA1c levels of 8 to 8.9 and a 0-0.3 variation of HbA1c levels
between screening and at baseline.
[0281] FIGS. 22-23 compare the changes in HbA1c levels for Groups 1
and 4 for those patients having a 0-0.3 variation of HbA1c levels
between screening and at t=0, with baseline HbA1c levels between 8
and 8.9. For this population, FIG. 22 depicts changes in HbA1c
levels across the 90 days for groups 1 and 4 and FIG. 23 depicts
the number of patients reaching specified HbA1c target levels at
the end of the study. FIG. 24 sets forth changes from baseline for
C-Peptide, FBG, Fructosamine, HbA1c, Insulin and Proinsulin.
[0282] Amongst the participants in the study there were no
significant adverse events, no episodes of severe hypoglycemia, or
weight gain amongst groups I-III. Incidents of mild to moderate
hypoglycemia and antibodies in groups 1-3 were comparable to those
found in group 4 (placebo).
* * * * *