U.S. patent application number 17/151980 was filed with the patent office on 2021-12-09 for lipid-based nanoparticles and use of same in optimized insulin dosing regimens.
The applicant listed for this patent is SDG, Inc.. Invention is credited to W. Blair Geho.
Application Number | 20210379160 17/151980 |
Document ID | / |
Family ID | 1000005787307 |
Filed Date | 2021-12-09 |
United States Patent
Application |
20210379160 |
Kind Code |
A1 |
Geho; W. Blair |
December 9, 2021 |
LIPID-BASED NANOPARTICLES AND USE OF SAME IN OPTIMIZED INSULIN
DOSING REGIMENS
Abstract
The invention provides methods of treating a subject having
diabetes mellitus and/or a metabolic derangement.
Inventors: |
Geho; W. Blair; (Chagrin
Falls, OH) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
SDG, Inc. |
Cleveland |
OH |
US |
|
|
Family ID: |
1000005787307 |
Appl. No.: |
17/151980 |
Filed: |
January 19, 2021 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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16846101 |
Apr 10, 2020 |
10918700 |
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17151980 |
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62988748 |
Mar 12, 2020 |
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62833228 |
Apr 12, 2019 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 38/28 20130101;
A61K 9/1277 20130101; A61P 3/10 20180101 |
International
Class: |
A61K 38/28 20060101
A61K038/28; A61P 3/10 20060101 A61P003/10; A61K 9/127 20060101
A61K009/127 |
Claims
1. A method of eliminating or minimizing events of iatrogenic
hyperinsulinemia or hypoglycemia in a subject having diabetes
mellitus or a metabolic derangement, wherein the subject is
originally administered an amount of bolus non-HDV insulin and an
amount of basal insulin such that the subject originally has
greater than about 8.5% HbA1c, the method comprising: administering
to the subject a bolus insulin HDV composition in place of the
original bolus non-HDV insulin, wherein the amount of insulin in
the bolus insulin HDV composition is lower than in the original
bolus non-HDV insulin; administering to the subject a basal
insulin; varying the administered amount of the bolus insulin HDV
composition and the administered amount of the basal insulin so as
to identify the optimized amount of the bolus insulin HDV
composition and the optimized amount of the basal insulin to be
administered to the subject to afford therapeutically effective
blood glucose control without, or with minimized, events of
iatrogenic hyperinsulinemia or hypoglycemia; wherein the bolus
insulin HDV composition comprises a lipid-based nanoparticle,
wherein the bolus insulin is dispersed within the nanoparticle,
wherein the nanoparticle is enclosed by a bipolar lipid membrane
comprising cholesterol, dicetyl phosphate, an amphipathic lipid,
and a hepatocyte receptor binding molecule; wherein the amphipathic
lipid comprises at least one selected from the group consisting of
1,2-distearoyl-sn-glycero-3-phosphocholine,
1,2-dipalmitoyl-sn-glycerol-[3-phospho-rac-(1-glycerol)],
1,2-distearoyl-sn-glycero-3-phosphoethanolamine,
1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-(succinyl),
1,2-dimyristoyl-sn-glycero-3-phosphate,
1,2-dimyristoyl-sn-glycero-3-phosphocholine, 1,2-di
stearoyl-sn-glycero-3-phosphate,
1,2-dipalmitoyl-sn-glycero-3-phosphate, and
1,2-dipalmitoyl-sn-glycero-3-phosphocholine; wherein the at least
one hepatocyte receptor binding molecule extends outward from the
nanoparticle; and wherein the size of the nanoparticle ranges from
about 10 nm to about 150 nm.
2. (canceled)
3. The method of claim 1, wherein the insulin ratio between the
optimized administered bolus insulin HDV composition and the
optimized administered basal insulin is equal to or lower than
1:1.
4-5. (canceled)
6. A method of eliminating or minimizing events of iatrogenic
hyperinsulinemia or hypoglycemia in a subject having diabetes
mellitus or a metabolic derangement, wherein the subject is
originally administered an amount of bolus non-HDV insulin and an
amount of basal insulin such that the subject originally has about
6.5-8.5% HbA1c, the method comprising: administering to the subject
a bolus insulin HDV composition in place of the original bolus
non-HDV insulin; administering to the subject a reducing amount of
basal insulin as compared to the amount of basal insulin originally
administered to the subject; and varying the administered amount of
the bolus insulin HDV composition so as to identify the optimized
amount of the bolus insulin HDV composition and the optimized
amount of the basal insulin to be administered to the subject such
that the diabetes is well controlled in the subject without, or
with minimized, events of iatrogenic hyperinsulinemia or
hypoglycemia; wherein the bolus insulin HDV composition comprises a
lipid-based nanoparticle, wherein the bolus insulin is dispersed
within the nanoparticle, wherein the nanoparticle is enclosed by a
bipolar lipid membrane comprising cholesterol, dicetyl phosphate,
an amphipathic lipid, and a hepatocyte receptor binding molecule;
wherein the amphipathic lipid comprises at least one selected from
the group consisting of 1,2-distearoyl-sn-glycero-3-phosphocholine,
1,2-dipalmitoyl-sn-glycerol-[3-phospho-rac-(1-glycerol)],
1,2-distearoyl-sn-glycero-3-phosphoethanolamine,
1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-(succinyl),
1,2-dimyristoyl-sn-glycero-3-phosphate,
1,2-dimyristoyl-sn-glycero-3-phosphocholine,
1,2-distearoyl-sn-glycero-3-phosphate,
1,2-dipalmitoyl-sn-glycero-3-phosphate, and
1,2-dipalmitoyl-sn-glycero-3-phosphocholine; wherein the at least
one hepatocyte receptor binding molecule extends outward from the
nanoparticle; and wherein the size of the nanoparticle ranges from
about 10 nm to about 150 nm.
7. The method of claim 6, wherein the subject before optimization
has about 80-100 mg/dL fasting blood sugar.
8. The method of claim 6, wherein at least one of the following
results is observed upon optimization: (a) the subject experiences
fewer hypoglycemia as compared to the treatment without HDV; (b)
the subject experiences weight loss as compared to the treatment
without HDV
9. (canceled)
10. The method of claim 6, wherein the bolus insulin HDV
composition further comprises a GLP-1 agonist or serotonin.
11. The method of claim 10, wherein the GLP-1 agonist comprises
liraglutide, semaglutide, or repaglinide.
12. The method of claim 6, wherein the basal insulin is formulated
in a composition comprising a lipid-based nanoparticle, wherein the
basal insulin is dispersed within the nanoparticle; wherein the
nanoparticle is enclosed by a bipolar lipid membrane comprising
cholesterol, dicetyl phosphate, an amphipathic lipid, and a
hepatocyte receptor binding molecule; wherein the amphipathic lipid
comprises at least one selected from the group consisting of
1,2-distearoyl-sn-glycero-3-phosphocholine,
1,2-dipalmitoyl-sn-glycerol-[3-phospho-rac-(1-glycerol)],
1,2-distearoyl-sn-glycero-3-phosphoethanolamine,
1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-(succinyl),
1,2-dimyristoyl-sn-glycero-3-phosphate,
1,2-dimyristoyl-sn-glycero-3-phosphocholine,
1,2-distearoyl-sn-glycero-3-phosphate,
1,2-dipalmitoyl-sn-glycero-3-phosphate, and
1,2-dipalmitoyl-sn-glycero-3-phosphocholine; wherein the at least
one hepatocyte receptor binding molecule extends outward from the
nanoparticle; and wherein the size of the nanoparticle ranges from
about 10 nm to about 150 nm.
13. The method of claim 6, wherein the basal insulin is
administered continuously to the subject over a period of at least
24 hours.
14. The method of claim 6, wherein the composition is administered
continuously to the subject using a pump.
15. (canceled)
16. The method of claim 6, wherein the membrane further comprises
at least one agent selected from the group consisting of a
stabilizer and stearoyl lysophosphatidylcholine.
17. The method of claim 16, wherein the stabilizer is selected from
the group consisting of m-cresol, benzyl alcohol, methyl
4-hydroxybenzoate, thiomersal, and butylated hydroxytoluene
(2,6-di-tert-butyl-4-methylphenol).
18. The method of claim 16, wherein the stabilizer ranges from
about 10% to about 25% (w/w) in the membrane, or the stearoyl
lysophosphatidylcholine ranges from about 5% to about 30% (w/w) in
the membrane.
19. The method of claim 6, wherein the insulin is covalently bound
to the nanoparticle or the insulin is not covalently bound to the
nanoparticle.
20. The method of claim 6, wherein the nanoparticle is suspended in
an aqueous solution comprising a free dissolved insulin that is not
dispersed within the nanoparticle.
21. The method of claim 20, wherein the nanoparticle-dispersed
insulin and the free dissolved insulin are independently selected
from the group consisting of insulin lispro, insulin aspart,
regular insulin, insulin glargine, insulin zinc, extended human
insulin zinc suspension, isophane insulin, human buffered regular
insulin, insulin glulisine, recombinant human regular insulin,
recombinant human insulin isophane, insulin detemir, biphasic human
insulin, and insulin deglude, and any combinations thereof.
22. The method of claim 6, wherein the amphipathic lipid comprises
at least one selected from the group consisting of
1,2-distearoyl-sn-glycero-3-phosphocholine,
1,2-dipalmitoyl-sn-glycero-3-phosphocholine,
1,2-dipalmitoyl-sn-glycero-3-[phospho-rac-(1-glycerol)],
1,2-distearoyl-sn-glycero-3-phosphoethanolamine, and
1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-(succinyl).
23. The method of claim 6, wherein the hepatocyte receptor binding
molecule comprises biotin.
24. The method of claim 23, wherein the biotin-containing
hepatocyte receptor binding molecule comprises at least one
selected from the group consisting of N-hydroxysuccinimide (NHS)
biotin; sulfo-NHS-biotin; N-hydroxysuccinimide long chain biotin;
sulfo-N-hydroxysuccinimide long chain biotin; D-biotin; biocytin;
sulfo-N-hydroxysuccinimide-S--S-biotin; biotin-BMCC; biotin-HPDP;
iodoacetyl-LC-biotin; biotin-hydrazide; biotin-LC-hydrazide;
biocytin hydrazide; biotin cadaverine; carboxybiotin; photobiotin;
p-aminobenzoyl biocytin trifluoroacetate; p-diazobenzoyl biocytin;
biotin DHPE (2,3-diacetoxypropyl
2-(5-((3aS,6aR)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamido)-
ethyl phosphate); biotin-X-DHPE (2,3-diacetoxypropyl
2-(6-(5-((3aS,6aR)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanami-
do)hexanamido) ethyl phosphate); 12-((biotinyl)amino)dodecanoic
acid; 12-((biotinyl)amino)dodecanoic acid succinimidyl ester;
S-biotinyl homocysteine; biocytin-X; biocytin x-hydrazide;
biotinethylenediamine; biotin-XL; biotin-X-ethylenediamine;
biotin-XX hydrazide; biotin-XX-SE; biotin-XX, SSE;
biotin-X-cadaverine; .alpha.-(t-BOC)biocytin;
N-(biotinyl)-N'-(iodoacetyl) ethylenediamine; DNP-X-biocytin-X-SE;
biotin-X-hydrazide; norbiotinamine hydrochloride;
3-(N-maleimidylpropionyl)biocytin; ARP; biotin-1-sulfoxide; biotin
methyl ester; biotin-maleimide; biotin-poly(ethyleneglycol) amine;
(+) biotin 4-amidobenzoic acid sodium salt; Biotin
2-N-acetylamino-2-deoxy-.beta.-D-glucopyranoside;
Biotin-.alpha.-D-N-acetylneuraminide; Biotin-.alpha.-L-fucoside;
Biotin lacto-N-bioside; Biotin-Lewis-A trisaccharide;
Biotin-Lewis-Y tetrasaccharide; Biotin-.alpha.-D-mannopyranoside;
and biotin 6-O-phospho-.alpha.-D-mannopyranoside.
25. The method of claim 23, wherein the biotin-containing
hepatocyte receptor binding molecule comprises at least one
selected from the group consisting of biotin DHPE and
biotin-X-DHPE.
26. The method of claim 6, wherein at least one applies: (a) the
cholesterol ranges from about 5% to about 25% (w/w) in the
membrane; (b) the dicetyl phosphate ranges from about 10% to about
25% (w/w) in the membrane; (c) the DSPC ranges from about 40% to
about 75% (w/w) in the membrane; (d) the hepatocyte receptor
binding molecule ranges from about 0.5% to about 10% (w/w) in the
membrane.
27. The method of claim 16, wherein the amount of the stearoyl
lysophosphatidylcholine in the membrane is about 5%-30% (w/w) of
the amount of DSPC in the membrane.
28. The method of claim 16, wherein the membrane comprises one of
the following: (a) cholesterol, dicetyl phosphate, DSPC, stearoyl
lysophosphatidylcholine, m-cresol, and at least one selected from
the group consisting of biotin DHPE and biotin-X-DHPE; (b)
cholesterol, dicetyl phosphate, DSPC, m-cresol, and at least one
selected from the group consisting of biotin DHPE and
biotin-X-DHPE; and (c) cholesterol, dicetyl phosphate, DSPC,
stearoyl lysophosphatidylcholine, and at least one selected from
the group consisting of biotin DHPE and biotin-X-DHPE.
29. The method of claim 16, wherein the membrane comprises
cholesterol, dicetyl phosphate, DSPC, stearoyl
lysophosphatidylcholine, m-cresol, and biotin DHPE in a % (w/w)
ratio selected from the group consisting of: (a) about
9.4:18.1:56.8:14.1:0.0:1.5; (b) about 7.7:15.0:58.6:0.0:17.4:1.3;
and (c) about 8.4:16.2:47.5:7.6:19.0:1.3.
30. (canceled)
31. The method of claim 1, wherein the bolus insulin HDV
composition further comprises a GLP-1 agonist or serotonin.
32. The method of claim 31, wherein the GLP-1 agonist comprises
liraglutide, semaglutide, or repaglinide.
33. The method of claim 1, wherein the basal insulin is
administered continuously to the subject over a period of at least
24 hours.
34. The method of claim 1, wherein the composition is administered
continuously to the subject using a pump.
35. The method of claim 1, wherein the membrane further comprises
at least one agent selected from the group consisting of a
stabilizer and stearoyl lysophosphatidylcholine.
36. The method of claim 35, wherein the stabilizer is selected from
the group consisting of m-cresol, benzyl alcohol, methyl
4-hydroxybenzoate, thiomersal, and butylated hydroxytoluene
(2,6-di-tert-butyl-4-methylphenol).
37. The method of claim 35, wherein the stabilizer ranges from
about 10% to about 25% (w/w) in the membrane, or the stearoyl
lysophosphatidylcholine ranges from about 5% to about 30% (w/w) in
the membrane.
38. The method of claim 1, wherein the insulin is covalently bound
to the nanoparticle or the insulin is not covalently bound to the
nanoparticle.
39. The method of claim 1, wherein the nanoparticle is suspended in
an aqueous solution comprising a free dissolved insulin that is not
dispersed within the nanoparticle.
40. The method of claim 39, wherein the nanoparticle-dispersed
insulin and the free dissolved insulin are independently selected
from the group consisting of insulin lispro, insulin aspart,
regular insulin, insulin glargine, insulin zinc, extended human
insulin zinc suspension, isophane insulin, human buffered regular
insulin, insulin glulisine, recombinant human regular insulin,
recombinant human insulin isophane, insulin detemir, biphasic human
insulin, and insulin deglude, and any combinations thereof.
41. The method of claim 1, wherein the amphipathic lipid comprises
at least one selected from the group consisting of
1,2-distearoyl-sn-glycero-3-phosphocholine,
1,2-dipalmitoyl-sn-glycero-3-phosphocholine,
1,2-dipalmitoyl-sn-glycero-3-[phospho-rac-(1-glycerol)],
1,2-distearoyl-sn-glycero-3-phosphoethanolamine, and
1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-(succinyl).
42. The method of claim 1, wherein the hepatocyte receptor binding
molecule comprises biotin.
43. The method of claim 42, wherein the biotin-containing
hepatocyte receptor binding molecule comprises at least one
selected from the group consisting of N-hydroxysuccinimide (NHS)
biotin; sulfo-NHS-biotin; N-hydroxysuccinimide long chain biotin;
sulfo-N-hydroxysuccinimide long chain biotin; D-biotin; biocytin;
sulfo-N-hydroxysuccinimide-S--S-biotin; biotin-BMCC; biotin-HPDP;
iodoacetyl-LC-biotin; biotin-hydrazide; biotin-LC-hydrazide;
biocytin hydrazide; biotin cadaverine; carboxybiotin; photobiotin;
p-aminobenzoyl biocytin trifluoroacetate; p-diazobenzoyl biocytin;
biotin DHPE (2,3-diacetoxypropyl
2-(5-((3aS,6aR)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamido)-
ethyl phosphate); biotin-X-DHPE (2,3-diacetoxypropyl
2-(6-(5-((3aS,6aR)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanami-
do)hexanamido) ethyl phosphate); 12-((biotinyl)amino)dodecanoic
acid; 12-((biotinyl)amino)dodecanoic acid succinimidyl ester;
S-biotinyl homocysteine; biocytin-X; biocytin x-hydrazide;
biotinethylenediamine; biotin-XL; biotin-X-ethylenediamine;
biotin-XX hydrazide; biotin-XX-SE; biotin-XX, SSE;
biotin-X-cadaverine; .alpha.-(t-BOC)biocytin;
N-(biotinyl)-N'-(iodoacetyl) ethylenediamine; DNP-X-biocytin-X-SE;
biotin-X-hydrazide; norbiotinamine hydrochloride;
3-(N-maleimidylpropionyl)biocytin; ARP; biotin-1-sulfoxide; biotin
methyl ester; biotin-maleimide; biotin-poly(ethyleneglycol) amine;
(+) biotin 4-amidobenzoic acid sodium salt; Biotin
2-N-acetylamino-2-deoxy-.beta.-D-glucopyranoside;
Biotin-.alpha.-D-N-acetylneuraminide; Biotin-.alpha.-L-fucoside;
Biotin lacto-N-bioside; Biotin-Lewis-A trisaccharide;
Biotin-Lewis-Y tetrasaccharide; Biotin-.alpha.-D-mannopyranoside;
and biotin 6-O-phospho-.alpha.-D-mannopyranoside.
44. The method of claim 42, wherein the biotin-containing
hepatocyte receptor binding molecule comprises at least one
selected from the group consisting of biotin DHPE and
biotin-X-DHPE.
45. The method of claim 1, wherein at least one applies: (a) the
cholesterol ranges from about 5% to about 25% (w/w) in the
membrane; (b) the dicetyl phosphate ranges from about 10% to about
25% (w/w) in the membrane; (c) the DSPC ranges from about 40% to
about 75% (w/w) in the membrane; (d) the hepatocyte receptor
binding molecule ranges from about 0.5% to about 10% (w/w) in the
membrane.
46. The method of claim 35, wherein the amount of the stearoyl
lysophosphatidylcholine in the membrane is about 5%-30% (w/w) of
the amount of DSPC in the membrane.
47. The method of claim 35, wherein the membrane comprises one of
the following: (a) cholesterol, dicetyl phosphate, DSPC, stearoyl
lysophosphatidylcholine, m-cresol, and at least one selected from
the group consisting of biotin DHPE and biotin-X-DHPE; (b)
cholesterol, dicetyl phosphate, DSPC, m-cresol, and at least one
selected from the group consisting of biotin DHPE and
biotin-X-DHPE; and (c) cholesterol, dicetyl phosphate, DSPC,
stearoyl lysophosphatidylcholine, and at least one selected from
the group consisting of biotin DHPE and biotin-X-DHPE.
48. The method of claim 35, wherein the membrane comprises
cholesterol, dicetyl phosphate, DSPC, stearoyl
lysophosphatidylcholine, m-cresol, and biotin DHPE in a % (w/w)
ratio selected from the group consisting of: (a) about
9.4:18.1:56.8:14.1:0.0:1.5; (b) about 7.7:15.0:58.6:0.0:17.4:1.3;
and (c) about 8.4:16.2:47.5:7.6:19.0:1.3.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a continuation of, and claims priority
to, U.S. patent application Ser. No. 16/846,101, filed Apr. 10,
2020, now allowed, which claims priority under 35 U.S.C. .sctn.
119(e) to U.S. Provisional Patent Applications No. 62/833,228,
filed Apr. 12, 2019, and No. 62/988,748, filed Mar. 12, 2020, all
of which are hereby incorporated herein by reference in their
entireties.
BACKGROUND OF THE INVENTION
[0002] Phospholipid nanoparticles of diameter lower than about 100
nm are often used as carriers to improve in vivo delivery of active
pharmaceutical ingredients (APIs), such as peptides and biogenic
amines. The nanoparticles' small particle size allows them to
easily cross membrane barriers. Further, nanoparticles may provide
rapid and specific delivery of APIs to desired cell surface
receptors, resulting in improved pharmacological action and need
for lower API doses. The targeted API delivery also leads to lower
toxicity, because of the API's reduced delivery to unwanted tissues
in the body.
[0003] An example of such nanoparticles is the hepatic delivery
vesicle (HDV), which comprises a hepatocyte-targeting component and
delivers APIs to hepatocyte receptors. In contrast, nanoparticles
without a hepatocyte-targeting components generally accumulate in
liver macrophages called Kupffer cells, along with other macrophage
cells in the body.
[0004] Diabetes mellitus, encompassing Type 1 and Type 2 forms, is
a disorder affecting large numbers of people worldwide. Diabetes
mellitus management comprises normalizing blood glucose levels in
the subject, and that may require multiple daily injections of an
insulin-based product. Despite the presence of various
insulin-based products on the market, there is still a need for
novel insulin-containing formulations that control glucose blood
levels in the subject over a wide period of time.
[0005] Certain medications approved for insulin-requiring diabetes
mellitus treatment comprise an insulin analog that is to be
administered subcutaneously, often as a time-release formulation.
Because of the abundance of insulin receptors in peripheral adipose
and muscle tissues, such administration releases the insulin analog
to peripheral tissues, but generally not to the liver. In one
aspect, proper insulin-requiring diabetes mellitus treatment
requires an insulin-based formulation in which a portion of the
dosed insulin is released to peripheral tissues throughout the day
and another portion of the dosed insulin is targeted for liver
delivery. Such need extends as well to other therapeutic agents for
which targeted liver delivery has advantageous therapeutic and/or
pharmacological properties.
[0006] There is thus an unmet need in the art for compositions and
methods for administering insulin to a subject, such that the
insulin is delivered to peripheral tissues as well as to the liver
of the subject. Such compositions and methods can be used to manage
blood glucose levels in Type 1 and Type 2 diabetic patients, as
well as patients with metabolic derangements, such as but not
limited to metabolic syndrome with elevated insulin levels,
steatosis, and/or steatohepatitis. The present invention meets this
need.
BRIEF SUMMARY OF THE INVENTION
[0007] The invention provides in one aspect a method of optimizing
the amount of bolus insulin and basal insulin to be administered to
a subject having diabetes mellitus, wherein the subject is
administered an amount of a bolus insulin HDV composition
comprising a lipid-based nanoparticle, wherein the bolus insulin is
dispersed within the nanoparticle, wherein the subject is further
administered an amount of basal insulin.
[0008] In certain embodiments, the method comprises varying the
administered amount of the bolus insulin HDV composition and the
administered amount of the basal insulin so as to identify the
optimized amount of the bolus insulin HDV composition and the
optimized amount of the basal insulin to be administered to the
subject to afford therapeutically effective blood glucose control
without significant hypoglycemia. In certain embodiments, the
nanoparticle is enclosed by a bipolar lipid membrane comprising
cholesterol, dicetyl phosphate, an amphipathic lipid, and a
hepatocyte receptor binding molecule. In certain embodiments, the
amphipathic lipid comprises at least one selected from the group
consisting of 1,2-distearoyl-sn-glycero-3-phosphocholine,
1,2-dipalmitoyl-sn-glycerol[3-phospho-rac-(1-glycerol)],
1,2-distearoyl-sn-glycero-3-phosphoethanolamine,
1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-(succinyl),
1,2-dimyristoyl-sn-glycero-3-phosphate,
1,2-dimyristoyl-sn-glycero-3-phosphocholine,
1,2-distearoyl-sn-glycero-3-phosphate,
1,2-dipalmitoyl-sn-glycero-3-phosphate, and
1,2-dipalmitoyl-sn-glycero-3-phosphocholine. In certain
embodiments, the at least one hepatocyte receptor binding molecule
extends outward from the nanoparticle. In certain embodiments, the
size of the nanoparticle ranges from about 10 nm to about 150
nm.
[0009] The invention provides in one aspect a method of optimizing
the amount of bolus insulin and basal insulin to be administered to
a subject having diabetes, wherein the subject is originally
administered an amount of bolus insulin and an amount of basal
insulin such that the diabetes is well controlled in the
subject.
[0010] In certain embodiments, the method comprises reducing the
amount of basal insulin administered to the subject and varying the
administered amount of a bolus insulin HDV composition so as to
identify the optimized amount of the bolus insulin HDV composition
and the optimized amount of the basal insulin to be administered to
the subject such that the diabetes is well controlled in the
subject. In certain embodiments, the bolus insulin HDV composition
comprises a lipid-based nanoparticle, wherein the bolus insulin is
dispersed within the nanoparticle. In certain embodiments, the
nanoparticle is enclosed by a bipolar lipid membrane comprising
cholesterol, dicetyl phosphate, an amphipathic lipid, and a
hepatocyte receptor binding molecule. In certain embodiments, the
amphipathic lipid comprises at least one selected from the group
consisting of 1,2-distearoyl-sn-glycero-3-phosphocholine,
1,2-dipalmitoyl-sn-glycerol-[3-phospho-rac-(1-glycerol)],
1,2-distearoyl-sn-glycero-3-phosphoethanolamine,
1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-(succinyl),
1,2-dimyristoyl-sn-glycero-3-phosphate,
1,2-dimyristoyl-sn-glycero-3-phosphocholine,
1,2-distearoyl-sn-glycero-3-phosphate,
1,2-dipalmitoyl-sn-glycero-3-phosphate, and
1,2-dipalmitoyl-sn-glycero-3-phosphocholine. In certain
embodiments, the at least one hepatocyte receptor binding molecule
extends outward from the nanoparticle. In certain embodiments, the
size of the nanoparticle ranges from about 10 nm to about 150
nm.
BRIEF DESCRIPTION OF THE DRAWINGS
[0011] For the purposes of illustrating the invention, there are
depicted in the drawings certain embodiments of the invention.
However, the invention is not limited to the precise arrangements
and instrumentalities of the embodiments depicted in the
drawings.
[0012] FIGS. 1A-1F illustrate changes in hypoglycemia, A1c, and
insulin by baseline A1c. p-Values indicate significance of
between-group differences at endpoint.
[0013] FIG. 2 illustrates selected results of continuous glucose
monitoring (CGM) studies in Example 1, in terms of % time that the
patient has blood glucose levels below 54 mg/dL vs. the patient's
A1c level.
[0014] FIG. 3 is an illustrative scheme for a Phase II dose
optimization study in lower A1C patients (6.5-8.5% A1C).
[0015] FIG. 4 illustrates median insulin dosing results for Example
3.
[0016] FIG. 5 illustrates hypoglycemic events per week (defined as
>15 Min CGM <54 mg/dL) for Example 3.
[0017] FIG. 6 illustrates hypoglycemic events per week (for day and
night) for Example 3.
[0018] FIG. 7 illustrates change from baseline (Visit 5) in weight
(kg) at Visit 11 for Example 3.
[0019] FIG. 8 illustrates change in mean glucose from optimized
baseline (baseline=mean of Visits 4&5) for Example 3.
[0020] FIG. 9 illustrates bolus: basal insulin ratios for Example
3.
[0021] FIG. 10 illustrates results for Example 3 relating to
hypoglycemic event results. In this study, a ninety-day unblinded
CGM, followed by an optimized standard of care, resulted in less
hypoglycemia events and 0.4% A1C reduction. When HDV was added to
unblinded CGM, subjects in both treatment groups achieved continued
decreases in hypoglycemia events, despite using more insulin
overall. Despite 10% or 40% reductions at Day 91 in basal insulin,
both treatment groups' basal dosing returned to baseline levels by
end of the study. Reductions in hypoglycemia during HDV treatment
did not result in increased overall glycemia.
[0022] FIG. 11 illustrates results for Example 3 relating to
hypoglycemic event results.
[0023] FIG. 12 illustrates results for Example 3 relating to
reduction in hypoglycemic events in relation to baseline. Based on
the studies using unblinded CGM and optimized standard of care, a
0.4% A1C improvement was observed after 90 days, with an about 11%
decrease in 24 hr and daytime hypoglycemic events, and with an
about 20% decrease in nighttime hypoglycemic events. The addition
of HDV to unblinded CGM allowed for additional 17% decrease in 24
hr hypoglycemic events, additional 6% decrease in daytime
hypoglycemic events, and additional 25% decrease in nighttime
hypoglycemic events. The addition of HDV therapy provided
hypoglycemia benefit despite the facts that subjects used slightly
higher overall insulin dosing and showed essentially no change in
A1C.
DETAILED DESCRIPTION OF THE INVENTION
[0024] The invention relates in part to the unexpected discovery
that HDV-insulin enables hepatic metabolism of ingested
carbohydrate (glucose), reducing the glucose load to peripheral
tissues, thus requiring an adjustment of basal doses of insulin so
that fasting hypoglycemia is reduced or eliminated. The present
invention provides, in one aspect, a new, physiologically adjusted
ratio of meal-time bolus HDV-insulin dose to the 24-hour basal
insulin, such as but not limited to degludec.
[0025] In certain embodiments, the use of the HDV-insulin
potentiates the effect on insulin in the subject, allowing for use
of lower amounts of insulin and thus avoiding iatrogenic
hyperinsulinemia and/or hypoglycemia in the subject.
[0026] In certain embodiments, the use of the HDV-insulin allows
for use of lower amounts of insulin, and thus reduce or eliminates
side effects associated with hyperinsulinemia (which can be derived
from use of large amounts of insulin), such as but not limited to
hypoglycemia, increased risk of polycystic ovary syndrome (PCOS),
increased synthesis of VLDL (hypertriglyceridemia), hypertension
(insulin increases sodium retention by the renal tubules), coronary
artery disease (increased insulin damages endothelial cells),
increased risk of cardiovascular disease, and/or weight gain and
lethargy.
[0027] In certain embodiments, the use of HDV-insulin allows for
efficacious, yet intermittent transient, engagement of hepatic
liver insulin receptors for the improvement of hepatic metabolic
function. In a non-limited, the HDV-insulin is administered to the
patient around meal time and allows for insulin to be delivered to
the hepatic insulin receptors during digestion. The HDV-insulin is
eventually removed from circulation through natural metabolic
processes and thus does not promote constitutive engagement of
hepatic liver insulin receptors, such as PEG-lispro, which remains
in circulation for prolonged periods of time, much after the need
to mealtime insulin has ceased.
[0028] Without wishing to be limited by any theory, the standard
treatment of a diabetic subject involves a 50:50 (or 1:1) ratio of
administered bolus insulin and basal insulin. Using HDV in at least
the bolus insulin allows for an insulin ratio that is closer to
physiological levels (such as, for example, using lower basal
insulin amounts).
[0029] In certain embodiments, the present invention provides a
method of optimizing the amount of bolus insulin and basal insulin
to be administered to a subject having diabetes mellitus. In other
embodiments, the subject is (initially) administered an amount of a
bolus insulin HDV composition comprising a lipid-based
nanoparticle, wherein the bolus insulin is dispersed within the
nanoparticle. In yet other embodiments, the subject is (initially)
further administered an amount of basal insulin. In yet other
embodiments, the method of the invention comprises varying the
administered amount of the bolus insulin HDV composition and the
administered amount of the basal insulin so as to identify the
amount of the bolus insulin HDV composition and the amount of the
basal insulin to be administered to the subject to afford
therapeutically effective blood glucose control without significant
hypoglycemia.
[0030] In certain embodiments, the present invention provides a
method of optimizing the amount of bolus insulin and basal insulin
to be administered to a subject having diabetes, wherein the
subject is originally administered an amount of bolus insulin and
an amount of basal insulin such that the diabetes is well
controlled in the subject. In other embodiments, the method
comprises reducing the amount of basal insulin administered to the
subject and varying the administered amount of a bolus insulin HDV
composition so as to identify the optimized amount of the bolus
insulin HDV composition and the optimized amount of the basal
insulin to be administered to the subject such that the diabetes is
well controlled in the subject. In yet other embodiments, the bolus
insulin HDV composition comprises a lipid-based nanoparticle,
wherein the bolus insulin is dispersed within the nanoparticle. In
yet other embodiments, the nanoparticle is enclosed by a bipolar
lipid membrane comprising cholesterol, dicetyl phosphate, an
amphipathic lipid, and a hepatocyte receptor binding molecule. In
yet other embodiments, the amphipathic lipid comprises at least one
selected from the group consisting of
1,2-distearoyl-sn-glycero-3-phosphocholine,
1,2-dipalmitoyl-sn-glycerol-[3-phospho-rac-(1-glycerol)],
1,2-distearoyl-sn-glycero-3-phosphoethanolamine,
1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-(succinyl),
1,2-dimyristoyl-sn-glycero-3-phosphate,
1,2-dimyristoyl-sn-glycero-3-phosphocholine,
1,2-distearoyl-sn-glycero-3-phosphate,
1,2-dipalmitoyl-sn-glycero-3-phosphate, and
1,2-dipalmitoyl-sn-glycero-3-phosphocholine. In yet other
embodiments, the at least one hepatocyte receptor binding molecule
extends outward from the nanoparticle. In yet other embodiments,
the size of the nanoparticle ranges from about 10 nm to about 150
nm.
[0031] In certain embodiments, the diabetes is diabetes
mellitus.
[0032] In certain embodiments, the subject has about 6.5-8.5% A1C.
In certain embodiments, the subject has 70-120 mg/dL fasting blood
sugar. In certain embodiments, the subject has 80-110 mg/dL fasting
blood sugar. In certain embodiments, the subject has 80-100 mg/dL
fasting blood sugar. In certain embodiments, the subject
experiences fewer hypoglycemia as compared to the treatment without
HDV. In certain embodiments, the reduction in the amount of bolus
insulin is about 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%,
50%, 55%, 60%, 65%, 70%, 75%, or 80%. In certain embodiments, the
reduction in the amount of bolus insulin ranges from about 10% to
about 40%. In certain embodiments, the subject experiences weight
loss as compared to the treatment without HDV.
[0033] In certain embodiments, the optimized insulin ratio between
the administered bolus insulin HDV composition (i.e., the amount of
bolus insulin in the HDV composition) and the administered basal
insulin depends in the severity of diabetes mellitus, which can be
measured in a non-limiting embodiment by hemoglobin A1c (HbA1c). In
certain embodiments, the optimized insulin ratio between the
administered bolus insulin HDV composition and the administered
basal insulin is equal to, or greater than, about 1:1 when the
subject has >8.5% HbA1c. In certain embodiments, the optimized
insulin ratio between the administered bolus insulin HDV
composition and the administered basal insulin is equal to, or
lower than, about 1:1 when the subject has <8.5% HbA1c. In
certain embodiments, the optimized insulin ratio between the
administered bolus insulin HDV composition and the administered
basal insulin is equal to, or greater than, about 1:1 when the
subject has <8.5% HbA1c. In certain embodiments, the optimized
insulin ratio between the administered bolus insulin HDV
composition and the administered basal insulin is equal to, or
lower than, about 1:1 when the subject has >8.5% HbA1c.
[0034] In certain embodiments, the subject has a HbA1c level equal
to or greater than about 10%. In certain embodiments, the subject
has a HbA1c level equal to or greater than about 9.9%. In certain
embodiments, the subject has a HbA1c level equal to or greater than
about 9.8%. In certain embodiments, the subject has a HbA1c level
equal to or greater than about 9.7%. In certain embodiments, the
subject has a HbA1c level equal to or greater than about 9.6%. In
certain embodiments, the subject has a HbA1c level equal to or
greater than about 9.5%. In certain embodiments, the subject has a
HbA1c level equal to or greater than about 9.4%. In certain
embodiments, the subject has a HbA1c level equal to or greater than
about 9.3%. In certain embodiments, the subject has a HbA1c level
equal to or greater than about 9.2%. In certain embodiments, the
subject has a HbA1c level equal to or greater than about 9.1%. In
certain embodiments, the subject has a HbA1c level equal to or
greater than about 9.0%. In certain embodiments, the subject has a
HbA1c level equal to or greater than about 8.9%. In certain
embodiments, the subject has a HbA1c level equal to or greater than
about 8.8%. In certain embodiments, the subject has a HbA1c level
equal to or greater than about 8.7%. In certain embodiments, the
subject has a HbA1c level equal to or greater than about 8.6%. In
certain embodiments, the subject has a HbA1c level equal to or
greater than about 8.5%. In certain embodiments, the subject has a
HbA1c level equal to or greater than about 8.4%. In certain
embodiments, the subject has a HbA1c level equal to or greater than
about 8.3%. In certain embodiments, the subject has a HbA1c level
equal to or greater than about 8.2%. In certain embodiments, the
subject has a HbA1c level equal to or greater than about 8.1%. In
certain embodiments, the subject has a HbA1c level equal to or
greater than about 8.0%. In certain embodiments, the subject has a
HbA1c level equal to or greater than about 7.9%. In certain
embodiments, the subject has a HbA1c level equal to or greater than
about 7.8%. In certain embodiments, the subject has a HbA1c level
equal to or greater than about 7.7%. In certain embodiments, the
subject has a HbA1c level equal to or greater than about 7.6%. In
certain embodiments, the subject has a HbA1c level equal to or
greater than about 7.5%. In certain embodiments, the subject has a
HbA1c level equal to or greater than about 7.4%. In certain
embodiments, the subject has a HbA1c level equal to or greater than
about 7.3%. In certain embodiments, the subject has a HbA1c level
equal to or greater than about 7.2%. In certain embodiments, the
subject has a HbA1c level equal to or greater than about 7.1%. In
certain embodiments, the subject has a HbA1c level equal to or
greater than about 7.0%. In certain embodiments, the subject has a
HbA1c level equal to or greater than about 6.9%. In certain
embodiments, the subject has a HbA1c level equal to or greater than
about 6.8%. In certain embodiments, the subject has a HbA1c level
equal to or greater than about 6.7%. In certain embodiments, the
subject has a HbA1c level equal to or greater than about 6.6%. In
certain embodiments, the subject has a HbA1c level equal to or
greater than about 6.5%.
[0035] In certain embodiments, the subject has a HbA1c level equal
to or lower than about 10%. In certain embodiments, the subject has
a HbA1c level equal to or lower than about 9.9%. In certain
embodiments, the subject has a HbA1c level equal to or lower than
about 9.8%. In certain embodiments, the subject has a HbA1c level
equal to or lower than about 9.7%. In certain embodiments, the
subject has a HbA1c level equal to or lower than about 9.6%. In
certain embodiments, the subject has a HbA1c level equal to or
lower than about 9.5%. In certain embodiments, the subject has a
HbA1c level equal to or lower than about 9.4%. In certain
embodiments, the subject has a HbA1c level equal to or lower than
about 9.3%. In certain embodiments, the subject has a HbA1c level
equal to or lower than about 9.2%. In certain embodiments, the
subject has a HbA1c level equal to or lower than about 9.1%. In
certain embodiments, the subject has a HbA1c level equal to or
lower than about 9.0%. In certain embodiments, the subject has a
HbA1c level equal to or lower than about 8.9%. In certain
embodiments, the subject has a HbA1c level equal to or lower than
about 8.8%. In certain embodiments, the subject has a HbA1c level
equal to or lower than about 8.7%. In certain embodiments, the
subject has a HbA1c level equal to or lower than about 8.6%. In
certain embodiments, the subject has a HbA1c level equal to or
lower than about 8.5%. In certain embodiments, the subject has a
HbA1c level equal to or lower than about 8.4%. In certain
embodiments, the subject has a HbA1c level equal to or lower than
about 8.3%. In certain embodiments, the subject has a HbA1c level
equal to or lower than about 8.2%. In certain embodiments, the
subject has a HbA1c level equal to or lower than about 8.1%. In
certain embodiments, the subject has a HbA1c level equal to or
lower than about 8.0%. In certain embodiments, the subject has a
HbA1c level equal to or lower than about 7.9%. In certain
embodiments, the subject has a HbA1c level equal to or lower than
about 7.8%. In certain embodiments, the subject has a HbA1c level
equal to or lower than about 7.7%. In certain embodiments, the
subject has a HbA1c level equal to or lower than about 7.6%. In
certain embodiments, the subject has a HbA1c level equal to or
lower than about 7.5%. In certain embodiments, the subject has a
HbA1c level equal to or lower than about 7.4%. In certain
embodiments, the subject has a HbA1c level equal to or lower than
about 7.3%. In certain embodiments, the subject has a HbA1c level
equal to or lower than about 7.2%. In certain embodiments, the
subject has a HbA1c level equal to or lower than about 7.1%. In
certain embodiments, the subject has a HbA1c level equal to or
lower than about 7.0%. In certain embodiments, the subject has a
HbA1c level equal to or lower than about 6.9%. In certain
embodiments, the subject has a HbA1c level equal to or lower than
about 6.8%. In certain embodiments, the subject has a HbA1c level
equal to or lower than about 6.7%. In certain embodiments, the
subject has a HbA1c level equal to or lower than about 6.6%. In
certain embodiments, the subject has a HbA1c level equal to or
greater than about 6.5%.
[0036] In certain embodiments, the optimized insulin ratio between
the administered bolus insulin HDV composition and the administered
basal insulin is about 1:0.1. In certain embodiments, the optimized
insulin ratio between the administered bolus insulin HDV
composition and the administered basal insulin is about 1:0.15. In
certain embodiments, the optimized insulin ratio between the
administered bolus insulin HDV composition and the administered
basal insulin is about 1:0.2. In certain embodiments, the optimized
insulin ratio between the administered bolus insulin HDV
composition and the administered basal insulin is about 1:0.25. In
certain embodiments, the optimized insulin ratio between the
administered bolus insulin HDV composition and the administered
basal insulin is about 1:0.3. In certain embodiments, the optimized
insulin ratio between the administered bolus insulin HDV
composition and the administered basal insulin is about 1:0.35. In
certain embodiments, the optimized insulin ratio between the
administered bolus insulin HDV composition and the administered
basal insulin is about 1:0.4. In certain embodiments, the optimized
insulin ratio between the administered bolus insulin HDV
composition and the administered basal insulin is about 1:0.45. In
certain embodiments, the optimized insulin ratio between the
administered bolus insulin HDV composition and the administered
basal insulin is about 1:0.5. In certain embodiments, the optimized
insulin ratio between the administered bolus insulin HDV
composition and the administered basal insulin is about 1:0.55. In
certain embodiments, the optimized insulin ratio between the
administered bolus insulin HDV composition and the administered
basal insulin is about 1:0.6. In certain embodiments, the optimized
insulin ratio between the administered bolus insulin HDV
composition and the administered basal insulin is about 1:0.65. In
certain embodiments, the optimized insulin ratio between the
administered bolus insulin HDV composition and the administered
basal insulin is about 1:0.7. In certain embodiments, the optimized
insulin ratio between the administered bolus insulin HDV
composition and the administered basal insulin is about 1:0.75. In
certain embodiments, the optimized insulin ratio between the
administered bolus insulin HDV composition and the administered
basal insulin is about 1:0.8. In certain embodiments, the optimized
insulin ratio between the administered bolus insulin HDV
composition and the administered basal insulin is about 1:0.85. In
certain embodiments, the optimized insulin ratio between the
administered bolus insulin HDV composition and the administered
basal insulin is about 1:0.9. In certain embodiments, the optimized
insulin ratio between the administered bolus insulin HDV
composition and the administered basal insulin is about 1:0.95. In
certain embodiments, the optimized insulin ratio between the
administered bolus insulin HDV composition and the administered
basal insulin is about 1:1. In certain embodiments, the optimized
insulin ratio between the administered bolus insulin HDV
composition and the administered basal insulin is about 1:1.05. In
certain embodiments, the optimized insulin ratio between the
administered bolus insulin HDV composition and the administered
basal insulin is about 1:1.1. In certain embodiments, the optimized
insulin ratio between the administered bolus insulin HDV
composition and the administered basal insulin is about 1:1.15. In
certain embodiments, the optimized insulin ratio between the
administered bolus insulin HDV composition and the administered
basal insulin is about 1:1.2. In certain embodiments, the optimized
insulin ratio between the administered bolus insulin HDV
composition and the administered basal insulin is about 1:1.25. In
certain embodiments, the optimized insulin ratio between the
administered bolus insulin HDV composition and the administered
basal insulin is about 1:1.3. In certain embodiments, the optimized
insulin ratio between the administered bolus insulin HDV
composition and the administered basal insulin is about 1:1.35. In
certain embodiments, the optimized insulin ratio between the
administered bolus insulin HDV composition and the administered
basal insulin is about 1:1.4. In certain embodiments, the optimized
insulin ratio between the administered bolus insulin HDV
composition and the administered basal insulin is about 1:1.45. In
certain embodiments, the optimized insulin ratio between the
administered bolus insulin HDV composition and the administered
basal insulin is about 1:1.5. In certain embodiments, the optimized
insulin ratio between the administered bolus insulin HDV
composition and the administered basal insulin is about 1:1.55. In
certain embodiments, the optimized insulin ratio between the
administered bolus insulin HDV composition and the administered
basal insulin is about 1:1.6. In certain embodiments, the optimized
insulin ratio between the administered bolus insulin HDV
composition and the administered basal insulin is about 1:1.65. In
certain embodiments, the optimized insulin ratio between the
administered bolus insulin HDV composition and the administered
basal insulin is about 1:1.7. In certain embodiments, the optimized
insulin ratio between the administered bolus insulin HDV
composition and the administered basal insulin is about 1:1.75. In
certain embodiments, the optimized insulin ratio between the
administered bolus insulin HDV composition and the administered
basal insulin is about 1:1.8. In certain embodiments, the optimized
insulin ratio between the administered bolus insulin HDV
composition and the administered basal insulin is about 1:1.85. In
certain embodiments, the optimized insulin ratio between the
administered bolus insulin HDV composition and the administered
basal insulin is about 1:1.9. In certain embodiments, the optimized
insulin ratio between the administered bolus insulin HDV
composition and the administered basal insulin is about 1:1.95. In
certain embodiments, the optimized insulin ratio between the
administered bolus insulin HDV composition and the administered
basal insulin is about 1:2. In certain embodiments, the optimized
insulin ratio between the administered bolus insulin HDV
composition and the administered basal insulin is about 1:2.05. In
certain embodiments, the optimized insulin ratio between the
administered bolus insulin HDV composition and the administered
basal insulin is about 1:2.1. In certain embodiments, the optimized
insulin ratio between the administered bolus insulin HDV
composition and the administered basal insulin is about 1:2.2. In
certain embodiments, the optimized insulin ratio between the
administered bolus insulin HDV composition and the administered
basal insulin is about 1:2.3. In certain embodiments, the optimized
insulin ratio between the administered bolus insulin HDV
composition and the administered basal insulin is about 1:2.4. In
certain embodiments, the optimized insulin ratio between the
administered bolus insulin HDV composition and the administered
basal insulin is about 1:2.5. In certain embodiments, the optimized
insulin ratio between the administered bolus insulin HDV
composition and the administered basal insulin is about 1:2.6. In
certain embodiments, the optimized insulin ratio between the
administered bolus insulin HDV composition and the administered
basal insulin is about 2.7. In certain embodiments, the optimized
insulin ratio between the administered bolus insulin HDV
composition and the administered basal insulin is about 1:2.8. In
certain embodiments, the optimized insulin ratio between the
administered bolus insulin HDV composition and the administered
basal insulin is about 1:2.9. In certain embodiments, the optimized
insulin ratio between the administered bolus insulin HDV
composition and the administered basal insulin is about 1:3. In
certain embodiments, the optimized insulin ratio between the
administered bolus insulin HDV composition and the administered
basal insulin is about 1:3.1. In certain embodiments, the optimized
insulin ratio between the administered bolus insulin HDV
composition and the administered basal insulin is about 1:3.2. In
certain embodiments, the optimized insulin ratio between the
administered bolus insulin HDV composition and the administered
basal insulin is about 1:3.3. In certain embodiments, the optimized
insulin ratio between the administered bolus insulin HDV
composition and the administered basal insulin is about 1:3.4. In
certain embodiments, the optimized insulin ratio between the
administered bolus insulin HDV composition and the administered
basal insulin is about 1:3.5. In certain embodiments, the optimized
insulin ratio between the administered bolus insulin HDV
composition and the administered basal insulin is about 1:3.6. In
certain embodiments, the optimized insulin ratio between the
administered bolus insulin HDV composition and the administered
basal insulin is about 1:3.7. In certain embodiments, the optimized
insulin ratio between the administered bolus insulin HDV
composition and the administered basal insulin is about 1:3.8. In
certain embodiments, the optimized insulin ratio between the
administered bolus insulin HDV composition and the administered
basal insulin is about 1:3.9. In certain embodiments, the optimized
insulin ratio between the administered bolus insulin HDV
composition and the administered basal insulin is about 1:4.0. In
certain embodiments, the optimized insulin ratio between the
administered bolus insulin HDV composition and the administered
basal insulin is about 1:4.1. In certain embodiments, the optimized
insulin ratio between the administered bolus insulin HDV
composition and the administered basal insulin is about 1:4.2. In
certain embodiments, the optimized insulin ratio between the
administered bolus insulin HDV composition and the administered
basal insulin is about 1:4.3. In certain embodiments, the optimized
insulin ratio between the administered bolus insulin HDV
composition and the administered basal insulin is about 1:4.4. In
certain embodiments, the optimized insulin ratio between the
administered bolus insulin HDV composition and the administered
basal insulin is about 1:4.5. In certain embodiments, the optimized
insulin ratio between the administered bolus insulin HDV
composition and the administered basal insulin is about 1:4.6. In
certain embodiments, the optimized insulin ratio between the
administered bolus insulin HDV composition and the administered
basal insulin is about 1:4.7. In certain embodiments, the optimized
insulin ratio between the administered bolus insulin HDV
composition and the administered basal insulin is about 1:4.8. In
certain embodiments, the optimized insulin ratio between the
administered bolus insulin HDV composition and the administered
basal insulin is about 1:4.9. In certain embodiments, the optimized
insulin ratio between the administered bolus insulin HDV
composition and the administered basal insulin is about 1:5.
[0037] In certain embodiments, the optimized insulin ratio between
the administered bolus insulin HDV composition and the administered
basal insulin is equal to, or greater than, about 1:0.1, about
1:0.15, about 1:0.2, about 1:0.25, about 1:0.3, about 1:0.35, about
1:0.4, about 1:0.45, about 1:0.5, about 1:0.55, about 1:0.6, about
1:0.65, about 1:0.7, about 1:0.75, about 1:0.8, about 1:0.85, about
1:0.9, about 1:0.95, about 1:1, about 1:1.05, about 1:1.1, about
1:1.15, about 1:1.2, about 1:1.25, about 1:1.3, about 1:1.35, about
1:1.4, about 1:1.45, about 1:1.5, about 1:1.55, about 1:1.6, about
1:1.65, about 1:1.7, about 1:1.75, about 1:1.8, about 1:1.85, about
1:1.9, about 1:1.95, about 1:2, about 1:2.05, about 1:2.1, about
1:2.2, about 1:2.3, about 1:2.4, about 1:2.5, about 1:2.6, about
2.7, about 1:2.8, about 1:2.9, about 1:3, about 1:3.1, about 1:3.2,
about 1:3.3, about 1:3.4, about 1:3.5, about 1:3.6, about 1:3.7,
about 1:3.8, about 1:3.9, about 1:4.0, about 1:4.1, about 1:4.2,
about 1:4.3, about 1:4.4, about 1:4.5, about 1:4.6, about 1:4.7,
about 1:4.8, about 1:4.9, and/or about 1:5.
[0038] In certain embodiments, the optimized insulin ratio between
the administered bolus insulin HDV composition and the administered
basal insulin is equal to, or lower than, about 1:0.1, about
1:0.15, about 1:0.2, about 1:0.25, about 1:0.3, about 1:0.35, about
1:0.4, about 1:0.45, about 1:0.5, about 1:0.55, about 1:0.6, about
1:0.65, about 1:0.7, about 1:0.75, about 1:0.8, about 1:0.85, about
1:0.9, about 1:0.95, about 1:1, about 1:1.05, about 1:1.1, about
1:1.15, about 1:1.2, about 1:1.25, about 1:1.3, about 1:1.35, about
1:1.4, about 1:1.45, about 1:1.5, about 1:1.55, about 1:1.6, about
1:1.65, about 1:1.7, about 1:1.75, about 1:1.8, about 1:1.85, about
1:1.9, about 1:1.95, about 1:2, about 1:2.05, about 1:2.1, about
1:2.2, about 1:2.3, about 1:2.4, about 1:2.5, about 1:2.6, about
2.7, about 1:2.8, about 1:2.9, about 1:3, about 1:3.1, about 1:3.2,
about 1:3.3, about 1:3.4, about 1:3.5, about 1:3.6, about 1:3.7,
about 1:3.8, about 1:3.9, about 1:4.0, about 1:4.1, about 1:4.2,
about 1:4.3, about 1:4.4, about 1:4.5, about 1:4.6, about 1:4.7,
about 1:4.8, about 1:4.9, and/or about 1:5.
[0039] In certain embodiments, the dose of insulin is per day
(daily).
[0040] In certain embodiments, the dose of insulin is about 0.01
units/kg. In certain embodiments, the dose of insulin is about 0.02
units/kg. In certain embodiments, the dose of insulin is about 0.03
units/kg. In certain embodiments, the dose of insulin is about 0.04
units/kg. In certain embodiments, the dose of insulin is about 0.05
units/kg. In certain embodiments, the dose of insulin is about 0.06
units/kg. In certain embodiments, the dose of insulin is about 0.07
units/kg. In certain embodiments, the dose of insulin is about 0.08
units/kg. In certain embodiments, the dose of insulin is about 0.09
units/kg. In certain embodiments, the dose of insulin is about 0.1
units/kg. In certain embodiments, the dose of insulin is about 0.15
units/kg. In certain embodiments, the dose of insulin is about 0.2
units/kg. In certain embodiments, the dose of insulin is about 0.25
units/kg. In certain embodiments, the dose of insulin is about 0.3
units/kg. In certain embodiments, the dose of insulin is about 0.35
units/kg. In certain embodiments, the dose of insulin is about 0.4
units/kg. In certain embodiments, the dose of insulin is about 0.45
units/kg. In certain embodiments, the dose of insulin is about 0.5
units/kg. In certain embodiments, the dose of insulin is about 0.55
units/kg. In certain embodiments, the dose of insulin is about 0.6
units/kg. In certain embodiments, the dose of insulin is about 0.65
units/kg. In certain embodiments, the dose of insulin is about 0.7
units/kg. In certain embodiments, the dose of insulin is about 0.75
units/kg. In certain embodiments, the dose of insulin is about 0.8
units/kg. In certain embodiments, the dose of insulin is about 0.85
units/kg. In certain embodiments, the dose of insulin is about 0.9
units/kg. In certain embodiments, the dose of insulin is about 0.95
units/kg. In certain embodiments, the dose of insulin is about 1
unit/kg. In certain embodiments, the dose of insulin is about 1.1
units/kg. In certain embodiments, the dose of insulin is about 1.2
units/kg. In certain embodiments, the dose of insulin is about 1.3
units/kg. In certain embodiments, the dose of insulin is about 1.4
units/kg. In certain embodiments, the dose of insulin is about 1.5
units/kg. In certain embodiments, the dose of insulin is about 1.6
units/kg. In certain embodiments, the dose of insulin is about 1.7
units/kg. In certain embodiments, the dose of insulin is about 1.8
units/kg. In certain embodiments, the dose of insulin is about 1.9
units/kg. In certain embodiments, the dose of insulin is about 2
units/kg. In certain embodiments, the dose of insulin is about 2.1
units/kg. In certain embodiments, the dose of insulin is about 2.2
units/kg. In certain embodiments, the dose of insulin is about 2.3
units/kg. In certain embodiments, the dose of insulin is about 2.4
units/kg. In certain embodiments, the dose of insulin is about 2.5
units/kg. In certain embodiments, the dose of insulin is about 2.6
units/kg. In certain embodiments, the dose of insulin is about 2.7
units/kg. In certain embodiments, the dose of insulin is about 2.8
units/kg. In certain embodiments, the dose of insulin is about 2.9
units/kg. In certain embodiments, the dose of insulin is about 3.0
units/kg. In certain embodiments, the dose of insulin is about 3.2
units/kg. In certain embodiments, the dose of insulin is about 3.4
units/kg. In certain embodiments, the dose of insulin is about 3.5
units/kg. In certain embodiments, the dose of insulin is about 3.6
units/kg. In certain embodiments, the dose of insulin is about 3.8
units/kg. In certain embodiments, the dose of insulin is about 4
units/kg. In certain embodiments, the dose of insulin is about 4.5
units/kg. In certain embodiments, the dose of insulin is about 5
units/kg. In certain embodiments, the dose of insulin is about 5.5
units/kg. In certain embodiments, the dose of insulin is about 6
units/kg. In certain embodiments, the dose of insulin is about 6.5
units/kg. In certain embodiments, the dose of insulin is about 7
units/kg. In certain embodiments, the dose of insulin is about 7.5
units/kg. In certain embodiments, the dose of insulin is about 8
units/kg. In certain embodiments, the dose of insulin is about 8.5
units/kg. In certain embodiments, the dose of insulin is about 9
units/kg. In certain embodiments, the dose of insulin is about 9.5
units/kg. In certain embodiments, the dose of insulin is about 10
units/kg. In certain embodiments, the dose of insulin is about 11
units/kg. In certain embodiments, the dose of insulin is about 12
units/kg. In certain embodiments, the dose of insulin is about 13
units/kg. In certain embodiments, the dose of insulin is about 14
units/kg. In certain embodiments, the dose of insulin is about 15
units/kg. In certain embodiments, the dose of insulin is about 16
units/kg. In certain embodiments, the dose of insulin is about 17
units/kg. In certain embodiments, the dose of insulin is about 18
units/kg. In certain embodiments, the dose of insulin is about 19
units/kg. In certain embodiments, the dose of insulin is about 20
units/kg.
[0041] In certain embodiments, the dose of insulin is greater than
about 0.01 units/kg, about 0.02 units/kg, about 0.03 units/kg,
about 0.04 units/kg, about 0.05 units/kg, about 0.06 units/kg,
about 0.07 units/kg, about 0.08 units/kg, about 0.09 units/kg,
about 0.1 units/kg, about 0.15 units/kg, about 0.2 units/kg, about
0.25 units/kg, about 0.3 units/kg, about 0.35 units/kg, about 0.4
units/kg, about 0.45 units/kg, about 0.5 units/kg, about 0.55
units/kg, about 0.6 units/kg, about 0.65 units/kg, about 0.7
units/kg, about 0.75 units/kg, about 0.8 units/kg, about 0.85
units/kg, about 0.9 units/kg, about 0.95 units/kg, about 1 unit/kg,
about 1.1 units/kg, about 1.2 units/kg, about 1.3 units/kg, about
1.4 units/kg, about 1.5 units/kg, about 1.6 units/kg, about 1.7
units/kg, about 1.8 units/kg, about 1.9 units/kg, about 2 units/kg,
about 2.1 units/kg, about 2.2 units/kg, about 2.3 units/kg, about
2.4 units/kg, about 2.5 units/kg, about 2.6 units/kg, about 2.7
units/kg, about 2.8 units/kg, about 2.9 units/kg, about 3.0
units/kg, about 3.2 units/kg, about 3.4 units/kg, about 3.5
units/kg, about 3.6 units/kg, about 3.8 units/kg, about 4 units/kg,
about 4.5 units/kg, about 5 units/kg, about 5.5 units/kg, about 6
units/kg, about 6.5 units/kg, about 7 units/kg, about 7.5 units/kg,
about 8 units/kg, about 8.5 units/kg, about 9 units/kg, about 9.5
units/kg, about 10 units/kg, about 11 units/kg, about 12 units/kg,
about 13 units/kg, about 14 units/kg, about 15 units/kg, about 16
units/kg, about 17 units/kg, about 18 units/kg, about 19 units/kg,
or about 20 units/kg.
[0042] In certain embodiments, the dose of insulin is lower than
about 0.01 units/kg, about 0.02 units/kg, about 0.03 units/kg,
about 0.04 units/kg, about 0.05 units/kg, about 0.06 units/kg,
about 0.07 units/kg, about 0.08 units/kg, about 0.09 units/kg,
about 0.1 units/kg, about 0.15 units/kg, about 0.2 units/kg, about
0.25 units/kg, about 0.3 units/kg, about 0.35 units/kg, about 0.4
units/kg, about 0.45 units/kg, about 0.5 units/kg, about 0.55
units/kg, about 0.6 units/kg, about 0.65 units/kg, about 0.7
units/kg, about 0.75 units/kg, about 0.8 units/kg, about 0.85
units/kg, about 0.9 units/kg, about 0.95 units/kg, about 1 unit/kg,
about 1.1 units/kg, about 1.2 units/kg, about 1.3 units/kg, about
1.4 units/kg, about 1.5 units/kg, about 1.6 units/kg, about 1.7
units/kg, about 1.8 units/kg, about 1.9 units/kg, about 2 units/kg,
about 2.1 units/kg, about 2.2 units/kg, about 2.3 units/kg, about
2.4 units/kg, about 2.5 units/kg, about 2.6 units/kg, about 2.7
units/kg, about 2.8 units/kg, about 2.9 units/kg, about 3.0
units/kg, about 3.2 units/kg, about 3.4 units/kg, about 3.5
units/kg, about 3.6 units/kg, about 3.8 units/kg, about 4 units/kg,
about 4.5 units/kg, about 5 units/kg, about 5.5 units/kg, about 6
units/kg, about 6.5 units/kg, about 7 units/kg, about 7.5 units/kg,
about 8 units/kg, about 8.5 units/kg, about 9 units/kg, about 9.5
units/kg, about 10 units/kg, about 11 units/kg, about 12 units/kg,
about 13 units/kg, about 14 units/kg, about 15 units/kg, about 16
units/kg, about 17 units/kg, about 18 units/kg, about 19 units/kg,
or about 20 units/kg.
[0043] In certain embodiments, the nanoparticles useful within the
invention are described in U.S. Patent Application Nos.
US20110135725 and US20090087479 and PCT Patent Application
Publication No. WO 2018/169954, all of which are incorporated
herein in their entireties by reference. In certain embodiments,
the reduced or minimal aggregation properties of the nanoparticle
of the invention improves its stability and pharmaceutical
developability as compared to nanoparticles of the prior art.
[0044] In certain embodiments, the lipid-based nanoparticle of the
invention is defined and/or enclosed by a bipolar lipid membrane.
In other embodiments, the nanoparticle of the invention comprises a
hepatocyte-targeting compound, which helps deliver the therapeutic
agent (such as, but not limited to, insulin) associated with,
and/or dispersed within, the nanoparticle to a hepatocyte. In yet
other embodiments, the nanoparticle of the invention is part of a
composition further comprising a "free" therapeutic agent, which is
not associated with, and/or dispersed within, the nanoparticle. The
nanoparticle, and any compositions comprising the same, can be
administered by any compatible and/or feasible routes, such as but
not limited to by injection (such as, for example, subcutaneously
and/or transdermally), inhalationally, buccally and/or orally, so
as to treat a subject that benefits from administration of the
therapeutic agent associated with, and/or dispersed within, the
nanoparticle, and/or of the "free" therapeutic agent, which is not
associated with, and/or dispersed within, the nanoparticle.
[0045] In certain embodiments, the therapeutic agent comprises
serotonin, or 5-hydroxytryptamine (5-HT), which is a monoamine
neurotransmitter.
[0046] In certain embodiments, the therapeutic agent comprises a
glucagon-like peptide-1 (GLP-1) agonist. GLP-1 is a potent incretin
hormone produced in the L-cells of the distal ileum and colon. In
the L-cells, GLP-1 is generated by tissue-specific
posttranslational processing of the proglucagon gene. Nutrients,
including glucose, fatty acids, and dietary fiber, are all known to
upregulate the transcription of the gene encoding GLP-1, and they
can stimulate the release of this hormone. The levels of GLP-1 rise
rapidly upon food ingestion. Nutrients, principally sugars and
fats, liberate GLP-1 and GLP-1-releasing factors, including
glucose-dependent insulinotropic peptide (GIP), gastrin-releasing
peptide, and selective neural regulators that also stimulate GLP-1
secretion. Non-limiting examples of GLP-1 agonists of interest are
liraglutide, semaglutide, and repaglinide.
[0047] Liposomes usually comprise amphipathic phospholipid
materials that form bilayer membranes that define and/or enclose
the liposomes. They can have a single membrane (unilamellar), or
multiple bilayers with a microscopic onion-like appearance.
Liposomes can be rather large, measuring several microns in
diameter. Liposomes generally have a spherical (or nearly
spherical) shape, wherein the intact surface has no available
"open" edges and thus cannot interact with other available "open"
edge liposome(s) to undergo particle aggregation.
[0048] In contrast, phospholipid nanoparticles with diameters equal
to or lower than about 200 nm have a restricted ability to bend
into a spherical configuration, which should in principle be their
thermodynamically stable structure. As a result, these low-diameter
nanoparticles do not form a perfectly spherical particle, but
rather a nearly planar sheet. Without wishing to be limited by any
theory, those nearly planar sheets can be described as "nanodiscs"
or "nanodisks" or "nanoFrisbees" or "bicelles." Such nanoparticles
have "open" edges in their membranes, and these "edges" promote
nanoparticle aggregation. As a result, in many instances the
nanoparticles are generated as discrete particles, which than
proceed to aggregate into larger, easily visible (wispy or
feather-like) floating particles. This phenomenon may hamper the
developability of the low-diameter nanoparticles as drug delivery
agents. In certain embodiments, unlike in the case of liposomes,
the API is not carried in the core volume of (or within) the
bicelles. In other embodiments, the API is attached and/or bound to
the membrane surface of the bicelles, either through a purely
physical interaction or a covalent linkage. In one aspect, the
present invention addresses this issue, providing compositions and
methods that allow for closing the "open" edges of the nearly
planar sheets (nanodiscs and/or nanoFrisbees) and thus minimizing
or suppressing their tendency to self-aggregate.
[0049] As described herein, in certain embodiments, the lipid-based
nanoparticles of the invention are useful as pharmaceutical
carriers, and do not form the wispy, feathery-like structures
described elsewhere herein. In certain embodiments, the
nanoparticles of the invention comprise certain amphipathic lipids
and/or certain organic molecules that enable the "open" edges of
the planar nanoparticle membranes to be changed in a way that
prevents aggregation of the nanoparticles.
[0050] In certain embodiments, appropriate closing of the "open"
edges of the lipid-based nanoparticle is promoted by replacing a
portion of distearoyl phosphatidylcholine [also known as
(S)-2,3-bis(stearoyloxy)propyl (2-(trimethylammonio)ethyl)
phosphate or DSPC, which comprises two C.sub.18 acyl groups
covalently linked to a glycerol backbone] with a C.sub.12-C.sub.24
acyl lysophosphatidylcholine [also known as C.sub.12-C.sub.24 acyl
lysolecithin, or 1-(C.sub.12-C.sub.24
acyl)-sn-glycero-3-phosphocholine, or
(S)-2-hydroxy-3-(C.sub.12-C.sub.24 acyloxy)propyl
(2-(trimethylammonio)ethyl) phosphate, which comprises a single
C.sub.12-C.sub.24 acyl group covalently linked to a glycerol
backbone]:
##STR00001##
[0051] In certain embodiments, appropriate closing of the "open"
edges of the lipid-based nanoparticle is promoted by replacing a
portion of distearoyl phosphatidylcholine [also known as
(S)-2,3-bis(stearoyloxy)propyl (2-(trimethylammonio)ethyl)
phosphate or DSPC, which comprises two C.sub.18 acyl groups
covalently linked to a glycerol backbone] with stearoyl
lysophosphatidylcholine [also known as
1-steroyl-sn-glycero-3-phosphocholine, or
(S)-2-hydroxy-3-(stearoyloxy)propyl (2-(trimethylammonio)ethyl)
phosphate, which comprises a single C.sub.18 acyl group covalently
linked to a glycerol backbone]:
##STR00002##
[0052] In certain embodiments, when incorporated into the membrane,
a C.sub.12-C.sub.24 acyl lysophosphatidylcholine (such as but not
limited to stearoyl lysophosphatidylcholine) prevents and/or
minimizes the aggregation that occurs when that compound is omitted
from the membrane. In other embodiments, the C.sub.12-C.sub.24 acyl
lysophosphatidylcholine (such as but not limited to stearoyl
lysophosphatidylcholine), with its single aliphatic chain, enables
closure of any existing membrane "edge" in the nanoparticle.
[0053] In certain embodiments, when incorporated into the membrane,
any of certain small molecule stabilizers or any salts and/or
solvates thereof, such as but not limited to m-cresol, benzyl
alcohol, methyl 4-hydroxybenzoate, thiomersal, and butylated
hydroxytoluene (also known as 2,6-di-tert-butyl-4-methylphenol),
prevents and/or minimizes the aggregation that occurs when that
compound is omitted from the membrane. In other embodiments, the
small molecule stabilizers or any salts and/or solvates thereof
enable closure of any existing membrane "edges" in the
nanoparticle.
[0054] In certain embodiments, when incorporated into the membrane,
any combinations of any of certain small molecule stabilizers or
any salts and/or solvates thereof, and the C.sub.12-C.sub.24 acyl
lysophosphatidylcholine, prevents and/or minimizes the aggregation
that occurs when that compound is omitted from the membrane.
Compositions
[0055] The invention provides lipid-based nanoparticles, and
compositions comprising the same. In certain embodiments, the
nanoparticle comprises, and/or is defined by, a bipolar lipid
membrane.
[0056] In certain embodiments, the membrane comprises cholesterol.
In other embodiments, the membrane comprises dicetyl phosphate. In
yet other embodiments, the membrane comprises an amphipathic lipid.
In yet other embodiments, the membrane comprises
1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC). In yet other
embodiments, the membrane comprises cholesterol, dicetyl phosphate,
and DSPC. In yet other embodiments, the membrane comprises a
hepatocyte receptor binding molecule.
[0057] In certain embodiments, the amphipathic lipid comprises at
least one selected from the group consisting of
1,2-distearoyl-sn-glycero-3-phosphocholine,
1,2-dipalmitoyl-sn-glycerol-3-phospho-rac-(1-glycerol)],
1,2-distearoyl-sn-glycero-3-phosphoethanolamine,
1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-(succinyl),
1,2-dimyristoyl-sn-glycero-3-phosphate,
1,2-dimyristoyl-sn-glycero-3-phosphocholine,
1,2-distearoyl-sn-glycero-3-phosphate,
1,2-dipalmitoyl-sn-glycero-3-phosphate, and
1,2-dipalmitoyl-sn-glycero-3-phosphocholine. In other embodiments,
the amphipathic lipid comprises at least one selected from the
group consisting of 1,2-distearoyl-sn-glycero-3-phosphocholine,
1,2-dipalmitoyl-sn-glycero-3-phosphocholine,
1,2-dipalmitoyl-sn-glycero-3-[phospho-rac-(1-glycerol)],
1,2-distearoyl-sn-glycero-3-phosphoethanolamine, and
1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-(succinyl).
[0058] In certain embodiments, the hepatocyte receptor binding
molecule comprises biotin. In other embodiments, the
biotin-containing hepatocyte receptor binding molecule comprises at
least one selected from the group consisting of
N-hydroxysuccinimide (NHS) biotin; sulfo-NHS-biotin;
N-hydroxysuccinimide long chain biotin; sulfo-N-hydroxysuccinimide
long chain biotin; D-biotin; biocytin;
sulfo-N-hydroxysuccinimide-S--S-biotin; biotin-BMCC; biotin-HPDP;
iodoacetyl-LC-biotin; biotin-hydrazide; biotin-LC-hydrazide;
biocytin hydrazide; biotin cadaverine; carboxybiotin; photobiotin;
p-aminobenzoyl biocytin trifluoroacetate; p-diazobenzoyl biocytin;
biotin DHPE (2,3-diacetoxypropyl
2-(5-((3aS,6aR)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamido)-
ethyl phosphate); biotin-X-DHPE (2,3-diacetoxypropyl
2-(6-(5-((3aS,6aR)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanami-
do)hexanamido) ethyl phosphate); 12-((biotinyl)amino)dodecanoic
acid; 12-((biotinyl)amino)dodecanoic acid succinimidyl ester;
S-biotinyl homocysteine; biocytin-X; biocytin x-hydrazide;
biotinethylenediamine; biotin-XL; biotin-X-ethylenediamine;
biotin-XX hydrazide; biotin-XX-SE; biotin-XX, SSE;
biotin-X-cadaverine; .alpha.-(t-BOC)biocytin;
N-(biotinyl)-N'-(iodoacetyl) ethylenediamine; DNP-X-biocytin-X-SE;
biotin-X-hydrazide; norbiotinamine hydrochloride;
3-(N-maleimidylpropionyl)biocytin; ARP; biotin-1-sulfoxide; biotin
methyl ester; biotin-maleimide; biotin-poly(ethyleneglycol) amine;
(+) biotin 4-amidobenzoic acid sodium salt; Biotin
2-N-acetylamino-2-deoxy-.beta.-D-glucopyranoside;
Biotin-.alpha.-D-N-acetylneuraminide; Biotin-.alpha.-L-fucoside;
Biotin lacto-N-bioside; Biotin-Lewis-A trisaccharide;
Biotin-Lewis-Y tetrasaccharide; Biotin-.alpha.-D-mannopyranoside;
and biotin 6-O-phospho-.alpha.-D-mannopyranoside.
[0059] In certain embodiments, the hepatocyte receptor binding
molecule is selected form the group consisting of
2,3-diacetoxypropyl
2-(5-((3aS,6aR)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamido)-
ethyl phosphate (biotin DHPE) and biotin-X-DHPE (2,3-diacetoxy
propyl
2-(6-(5-((3aS,6aR)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanami-
do) hexanamido)ethyl phosphate).
[0060] In certain embodiments, the cholesterol ranges from about 5%
to about 25% (w/w) in the membrane. In other embodiments, the
cholesterol is present in the membrane at a concentration of about
5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, 10%, 10.5%, 11%,
11.5%, 12%, 12.5%, 13%, 13.5%, 14%, 14.5%, 15%, 15.5%, 16%, 16.5%,
17%, 17.5%, 18%, 18.5%, 19%, 19.5%, 20%, 20.5%, 21%, 21.5%, 22%,
22.5%, 23%, 23.5%, 24%, 24.5%, or 25% (w/w).
[0061] In certain embodiments, the dicetyl phosphate ranges from
about 10% to about 25% (w/w) in the membrane. In other embodiments,
the dicetyl phosphate is present in the membrane at a concentration
of about 10%, 10.5%, 11%, 11.5%, 12%, 12.5%, 13%, 13.5%, 14%,
14.5%, 15%, 15.5%, 16%, 16.5%, 17%, 17.5%, 18%, 18.5%, 19%, 19.5%,
20%, 20.5%, 21%, 21.5%, 22%, 22.5%, 23%, 23.5%, 24%, 24.5%, or 25%
(w/w).
[0062] In certain embodiments, the DSPC ranges from about 40% to
about 75% (w/w) in the membrane. In other embodiments, the DSPC is
present in the membrane at a concentration of about 40%, 41%, 42%,
43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%,
56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%,
69%, 70%, 71%, 72%, 73%, 74%, or 75% (w/w).
[0063] In certain embodiments, the hepatocyte receptor binding
molecule ranges from about 0.5% to about 10% (w/w) in the membrane.
In other embodiments, the hepatocyte receptor binding molecule is
present in the membrane at a concentration of about 0.5%, 0.6%,
0.7%, 0.8%, 0.9%, 1.0%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%,
1.8%, 1.9%, 2.0%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%,
2.9%, 3.0%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3.7%, 3.8%, 3.9%,
4.0%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, or
10% (w/w).
[0064] In certain embodiments, the membrane comprises at least one
compound selected from the group consisting of a stabilizer and a
C.sub.12-C.sub.24 acyl lysophosphatidylcholine.
[0065] In certain embodiments, the membrane further comprises a
C.sub.12-C.sub.24 acyl lysophosphatidylcholine. In other
embodiments, the membrane further comprises stearoyl
lysophosphatidylcholine.
[0066] In certain embodiments, the membrane further comprises
m-cresol.
[0067] In certain embodiments, the stabilizer is selected from the
group consisting of m-cresol, benzyl alcohol, methyl
4-hydroxybenzoate, thiomersal, and butylated hydroxytoluene
(2,6-di-tert-butyl-4-methylphenol).
[0068] In certain embodiments, the stabilizer ranges from about 10%
to about 25% (w/w) in the membrane. In other embodiments, the
stabilizer is present in the membrane at a concentration of about
10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%,
23%, 24%, or 25% (w/w).
[0069] In certain embodiments, the m-cresol ranges from about 10%
to about 25% (w/w) in the membrane. In other embodiments, the
m-cresol is present in the membrane at a concentration of about
10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%,
23%, 24%, or 25% (w/w).
[0070] In certain embodiments, the C.sub.12-C.sub.24
lysophosphatidylcholine ranges from about 5% to about 30% (w/w) in
the membrane. In other embodiments, the C.sub.12-C.sub.24
lysophosphatidylcholine ranges from about 1% to about 30% (w/w) in
the membrane. In yet other embodiments, the C.sub.12-C.sub.24
lysophosphatidylcholine is present in the membrane at a
concentration of about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%,
11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%,
24%, 25%, 26%, 27%, 28%, 29% or 30% (w/w).
[0071] In certain embodiments, the stearoyl lysophosphatidylcholine
ranges from about 5% to about 30% (w/w) in the membrane. In other
embodiments, the stearoyl lysophosphatidylcholine ranges from about
1% to about 30% (w/w) in the membrane. In yet other embodiments,
the stearoyl lysophosphatidylcholine is present in the membrane at
a concentration of about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%,
11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%,
24%, 25%, 26%, 27%, 28%, 29% or 30% (w/w).
[0072] In certain embodiments, the amount of the C.sub.12-C.sub.24
lysophosphatidylcholine in the membrane is about 1% to about 30%
(w/w) of the amount of DSPC in the membrane. In yet other
embodiments, the amount of the C.sub.12-C.sub.24
lysophosphatidylcholine in the membrane is about 1%, 2%, 3%, 4%,
5%, 6%, 7%, 8%, 9%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%,
15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%,
28%, 29% (w/w) or 30% (w/w) of the amount of DSPC in the
membrane.
[0073] In certain embodiments, the amount of the C.sub.12-C.sub.24
lysophosphatidylcholine in the membrane is about 1 mole % to about
50 mole % of the amount of DSPC in the membrane. In yet other
embodiments, the amount of the C.sub.12-C.sub.24
lysophosphatidylcholine in the membrane is about 1, 2, 3, 4, 5, 6,
7, 8, 9, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20,
21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37,
38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50 mole % of the
amount of DSPC in the membrane.
[0074] In certain embodiments, the amount of the stearoyl
lysophosphatidylcholine in the membrane is about 1% to about 30%
(w/w) of the amount of DSPC in the membrane. In yet other
embodiments, the amount of the stearoyl lysophosphatidylcholine in
the membrane is about 1%, 6%, 7%, 8%, 9%, 10%-11%, 12%, 13%, 14%,
15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%,
28%, 29% or 30% (w/w) of the amount of DSPC in the membrane.
[0075] In certain embodiments, the amount of the stearoyl
lysophosphatidylcholine in the membrane is about 1 mole % to about
50 mole % of the amount of DSPC in the membrane. In yet other
embodiments, the amount of the stearoyl lysophosphatidylcholine in
the membrane is about 1, 2, 3, 4, 5, 6, 7, 8, 9, 5, 6, 7, 8, 9, 10,
11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27,
28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44,
45, 46, 47, 48, 49, or 50 mole % of the amount of DSPC in the
membrane.
[0076] In certain embodiments, the membrane comprises cholesterol,
dicetyl phosphate, DSPC, stearoyl lysophosphatidylcholine,
m-cresol, and at least one selected from the group consisting of
biotin DHPE and biotin-X-DHPE. In other embodiments, the membrane
comprises cholesterol, dicetyl phosphate, DSPC, stearoyl
lysophosphatidylcholine, m-cresol, and biotin DHPE.
[0077] In certain embodiments, the membrane comprises cholesterol,
dicetyl phosphate, DSPC, m-cresol, and at least one selected from
the group consisting of biotin DHPE and biotin-X-DHPE. In other
embodiments, the membrane comprises cholesterol, dicetyl phosphate,
DSPC, m-cresol, and biotin DHPE.
[0078] In certain embodiments, the membrane comprises cholesterol,
dicetyl phosphate, DSPC, stearoyl lysophosphatidylcholine, and at
least one selected from the group consisting of biotin DHPE and
biotin-X-DHPE. In other embodiments, the membrane comprises
cholesterol, dicetyl phosphate, DSPC, stearoyl
lysophosphatidylcholine, and biotin DHPE.
[0079] In certain embodiments, the stabilizer is contacted with the
membrane, and/or the lipid components that assemble to form the
membrane (such as, but not limited to, cholesterol, dicetyl
phosphate, DSPC, C.sub.12-C.sub.24 lysophosphatidylcholine if
present, and biotin DHPE), at a (w/w) ratio of the membrane to the
stabilizer ranging from about 1:1 to about 1:30. In other
embodiments, the stabilizer is contacted with the membrane, and/or
the lipid components that assemble to form the membrane, at a (w/w)
ratio of the membrane to the stabilizer of about 1:1, 1:1.5, 1:2,
1:2.5, 1:3, 1:3.5, 1:4, 1:4.5, 1:5, 1:5.5, 1:6, 1:6.5, 1:7, 1:7.5,
1:8, 1:8.5, 1:9, 1:9.5, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16,
1:17, 1:18, 1:19, 1:20, 1:21, 1;22, 1:23, 1:24, 1:25, 1:26, 1:27,
1:28, 1:29 or 1:30.
[0080] In certain embodiments, the m-cresol is contacted with the
membrane, and/or the lipid components that assemble to form the
membrane (such as, but not limited to, cholesterol, dicetyl
phosphate, DSPC, C.sub.12-C.sub.24 lysophosphatidylcholine if
present, and biotin DHPE), at a (w/w) ratio of the membrane to the
stabilizer ranging from about 1:1 to about 1:30. In other
embodiments, the m-cresol is contacted with the membrane, and/or
the lipid components that assemble to form the membrane, at a (w/w)
ratio of the membrane to the stabilizer of about 1:1, 1:1.5, 1:2,
1:2.5, 1:3, 1:3.5, 1:4, 1:4.5, 1:5, 1:5.5, 1:6, 1:6.5, 1:7, 1:7.5,
1:8, 1:8.5, 1:9, 1:9.5, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16,
1:17, 1:18, 1:19, 1:20, 1:21, 1;22, 1:23, 1:24, 1:25, 1:26, 1:27,
1:28, 1:29 or 1:30.
[0081] In certain embodiments, the membrane comprises cholesterol,
dicetyl phosphate, DSPC, stearoyl lysophosphatidylcholine,
m-cresol, and biotin DHPE, in a % (w/w) ratio of about
9.4:18.1:56.8:14.1:0.0:1.5.
[0082] In certain embodiments, the membrane comprises cholesterol,
dicetyl phosphate, DSPC, stearoyl lysophosphatidylcholine, and
biotin DHPE, in a % (w/w) ratio of about
9.4:18.1:56.8:14.1:1.5.
[0083] In certain embodiments, the membrane comprises cholesterol,
dicetyl phosphate, DSPC, stearoyl lysophosphatidylcholine,
m-cresol, and biotin DHPE, in a % (w/w) ratio of about
7.7:15.0:58.6:0.0:17.4:1.3.
[0084] In certain embodiments, the membrane comprises cholesterol,
dicetyl phosphate, DSPC, and biotin DHPE, in a % (w/w) ratio of
about 9.3:18.2:71.0:1.5.
[0085] In certain embodiments, the membrane comprises cholesterol,
dicetyl phosphate, DSPC, stearoyl lysophosphatidylcholine,
m-cresol, and biotin DHPE, in a % (w/w) ratio of about
8.4:16.2:47.5:7.6:19.0:1.3.
[0086] In certain embodiments, the membrane comprises cholesterol,
dicetyl phosphate, DSPC, stearoyl lysophosphatidylcholine, and
biotin DHPE, in a % (w/w) ratio of about 10.4:20:58.6:9.4:1.6.
[0087] In certain embodiments, the at least one hepatocyte receptor
binding molecule extends outward from the nanoparticle.
[0088] The invention should not be construed to be limited to the
constructs described and/or exemplified herein. Rather, the
invention provides methods of stabilizing and/or preventing
aggregation of liposomes and other lipid-based nanoparticles,
wherein the membrane is contacted with at least one selected from
the group consisting of a stabilizer and a C.sub.12-C.sub.24 acyl
lysophosphatidylcholine. In certain embodiments, the contacting
removes or minimizes any "free" edges in the membrane that lead to
aggregation of the liposomes and other lipid-based
nanoparticles.
[0089] In certain embodiments, the stabilizer is selected from the
group consisting of m-cresol, benzyl alcohol, methyl
4-hydroxybenzoate, thiomersal, and butylated hydroxytoluene. In
other embodiments, the stabilizer, such as but not limited to
m-cresol, ranges from about 10% to about 25% (w/w) in the membrane.
In yet other embodiments, the stabilizer, such as but not limited
to m-cresol, is present in the membrane at a concentration of about
10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%,
23%, 24%, or 25% (w/w).
[0090] In certain embodiments, the C.sub.12-C.sub.24
lysophosphatidylcholine, such as but not limited to stearoyl
lysophosphatidylcholine, ranges from about 5% to about 30% (w/w) in
the membrane. In other embodiments, the C.sub.12-C.sub.24
lysophosphatidylcholine, such as but not limited to stearoyl
lysophosphatidylcholine, ranges from about 1% to about 30% (w/w) in
the membrane. In yet other embodiments, the C.sub.12-C.sub.24
lysophosphatidylcholine, such as but not limited to stearoyl
lysophosphatidylcholine, is present in the membrane at a
concentration of about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%,
11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%,
24%, 25%, 26%, 27%, 28%, 29% or 30% (w/w).
[0091] In certain embodiments, the membrane comprises at least one
amphipathic lipid selected to from the group consisting of
1,2-distearoyl-sn-glycero-3-phosphocholine,
1,2-dipalmitoyl-sn-glycerol-[3-phospho-rac-(1-glycerol)],
1,2-distearoyl-sn-glycero-3-phosphoethanolamine,
1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-(succinyl),
1,2-dimyristoyl-sn-glycero-3-phosphate,
1,2-dimyristoyl-sn-glycero-3-phosphocholine,
1,2-distearoyl-sn-glycero-3-phosphate,
1,2-dipalmitoyl-sn-glycero-3-phosphate, and
1,2-dipalmitoyl-sn-glycero-3-phosphocholine. In other embodiments,
the amphipathic lipid is at least one selected from the group
consisting of 1,2-distearoyl-sn-glycero-3-phosphocholine,
1,2-dipalmitoyl-sn-glycero-3-phosphocholine,
1,2-dipalmitoyl-sn-glycero-3-[phospho-rac-(1-glycerol)],
1,2-distearoyl-sn-glycero-3-phosphoethanolamine, and
1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-(succinyl).
[0092] In certain embodiments, the amount of the C.sub.12-C.sub.24
lysophosphatidylcholine in the membrane is about 1%-30% (w/w) of
the amount of the at least one amphipathic lipid in the membrane.
In yet other embodiments, the amount of the C.sub.12-C.sub.24
lysophosphatidylcholine in the membrane is about 1%, 2%, 3%, 4%,
5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%,
19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29% or 30% (w/w)
of the amount of the at least one amphipathic lipid in the
membrane.
[0093] In certain embodiments, the amount of the C.sub.12-C.sub.24
lysophosphatidylcholine in the membrane is about 1 mole % to about
50 mole % of the amount of the at least one amphipathic lipid in
the membrane. In yet other embodiments, the amount of the
C.sub.12-C.sub.24 lysophosphatidylcholine in the membrane is about
1, 2, 3, 4, 5, 6, 7, 8, 9, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15,
16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32,
33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49,
or 50 mole % of the amount of the at least one amphipathic lipid in
the membrane.
[0094] In certain embodiments, the stabilizer, such as but not
limited to m-cresol, is contacted with the membrane, and/or the
lipid components that assemble to form the membrane, at a (w/w)
ratio ranging from about 1:1 to about 1:30. In other embodiments,
the stabilizer, such as but not limited to m-cresol, is contacted
with the membrane, and/or the lipid components that assemble to
form the membrane, at a (w/w) ratio of about 1:1, 1:1.5, 1:2,
1:2.5, 1:3, 1:3.5, 1:4, 1:4.5, 1:5, 1:5.5, 1:6, 1:6.5, 1:7, 1:7.5,
1:8, 1:8.5, 1:9, 1:9.5, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16,
1:17, 1:18, 1:19, 1:20, 1:21, 1;22, 1:23, 1:24, 1:25, 1:26, 1:27,
1:28, 1:29 or 1:30.
[0095] In certain embodiments, the size of the nanoparticle ranges
from about 10 nm to about 150 nm. In other embodiments, the size of
the nanoparticle is about 10 nm, 20 nm, 30 nm, 40 nm, 50 nm, 60 nm,
70 nm, 80 nm, 90 nm, 100 nm, 110 nm, 120 nm, 130 nm, 140 nm or 150
nm.
[0096] In certain embodiments, a therapeutic agent (such as, but
not limited to, insulin) is dispersed within and/or adsorbed onto
the nanoparticle. In other embodiments, the therapeutic agent is
covalently bound to the nanoparticle. In yet other embodiments, the
therapeutic agent is not covalently bound to the nanoparticle.
[0097] In certain embodiments, the therapeutic agent comprises at
least one selected from the group consisting of insulin, insulin
analogs, GLP-1 agonist, amylin, interferon, parathyroid hormone,
calcitonin, serotonin, serotonin agonist, serotonin reuptake
inhibitor, human growth hormone, GIP, anti-GIP monoclonal antibody,
metformin, bromocriptine, dopamine, glucagon, and GLP-1. In other
embodiments, the therapeutic agent is insulin.
[0098] In certain embodiments, the nanoparticle is suspended in an
aqueous solution comprising a free dissolved therapeutic agent that
is not dispersed within the nanoparticle.
[0099] In certain embodiments, the nanoparticle-dispersed insulin
and the free dissolved insulin are independently selected from the
group consisting of insulin lispro, insulin aspart (including
FIASP.RTM., Novo Nordisk), regular insulin, insulin glargine,
insulin zinc, extended human insulin zinc suspension, isophane
insulin, human buffered regular insulin, insulin glulisine,
recombinant human regular insulin, recombinant human insulin
isophane, insulin detemir, biphasic human insulin, and insulin
degludec (including TRESIBA.RTM., Novo Nordisk).
[0100] In certain embodiments, the lipid further comprises
cellulose acetate phthalate. In other embodiments, the cellulose
acetate phthalate is at least partially bound to the therapeutic
agent dispersed within the nanoparticle.
[0101] In certain embodiments, at least one charged organic
molecule is bound to the therapeutic agent dispersed within the
nanoparticle. In other embodiments, the charged organic molecule is
at least one selected from the group consisting of protamines,
polylysine, poly (arg-pro-thr)n in a mole ratio of 1:1:1, poly
(DL-Ala-poly-L-lys)n in a mole ratio of 6:1, histones, sugar
polymers comprising a primary amino group, polynucleotides with
primary amino groups, proteins comprising amino acid residues with
carboxyl (COO.sup.-) or sulfhydral (S.sup.-) functional groups, and
acidic polymers (such as sugar polymers containing carboxyl
groups).
[0102] In certain embodiments, the nanoparticle of the invention,
and compositions comprising the same, help deliver the therapeutic
agent dispersed therewithin to the hepatocytes in the liver.
[0103] In certain embodiments, the compositions of the invention
comprise an effective dose of a hepatocyte targeted pharmaceutical
composition that combines free therapeutic drug (such as, but not
limited to, insulin) and therapeutic drug associated with the
lipid-based nanoparticle of the invention. The combination of free
therapeutic drug and therapeutic drug associated with the
lipid-based nanoparticle creates a dynamic equilibrium process
between the two forms of therapeutic drug that occurs in vivo to
help control the movement of free therapeutic drug to the receptor
sites of hormonal action. In the case of insulin as the therapeutic
drug, those receptor sites are the muscle and adipose tissues of a
diabetic patient. Hepatocyte targeted therapeutic drug is also
delivered to the liver of a patient over a different designated
time period than free therapeutic drug, thereby introducing new
pharmacodynamic profiles of therapeutic drug when the therapeutic
drug remains associated with the nanoparticle and/or when free
therapeutic drug is released from the nanoparticle. In addition, a
portion of therapeutic drug that is associated with the
nanoparticle is targeted to the liver. In the case of insulin as
the therapeutic drug, the new pharmacodynamic profile of the
product provides not only basal insulin for peripheral tissues, but
also meal-time hepatic therapeutic drug stimulation for the
management of hepatic glucose storage during a meal. Free insulin
is released from the site of administration and is distributed
throughout the body. Insulin associated with the lipid-based
nanoparticle is delivered to the liver. The rate of release of
insulin associated with the nanoparticle is different than the rate
of release of free insulin from the site of administration. These
different release rates of insulin delivery, combined with the
targeted delivery of insulin associated with the nanoparticle to
the liver, provide for the normalization of glucose concentrations
in patients with Type 1 and Type 2 diabetes mellitus, as well as
patients with metabolic derangements, such as but not limited to
metabolic syndrome with elevated insulin levels, steatosis, and/or
steatohepatitis. In certain embodiments, the hepatocyte targeted
composition comprises any therapeutically effective insulin or
insulin derivative or analog, or any combination of two or more
types of insulin or insulin derivative or analog.
[0104] Compounds described herein also include isotopically labeled
compounds wherein one or more atoms is replaced by an atom having
the same atomic number, but an atomic mass or mass number different
from the atomic mass or mass number usually found in nature.
Examples of isotopes suitable for inclusion in the compounds
described herein include and are not limited to .sup.2H, .sup.3H,
.sup.11C, .sup.13C, .sup.14C, .sup.36Cl, .sup.18F, .sup.123I,
.sup.125I, .sup.13N, .sup.15N, .sup.15O, .sup.17O, .sup.18O,
.sup.32P, and .sup.35S. In certain embodiments, isotopically
labeled compounds are useful in drug and/or substrate tissue
distribution studies. In other embodiments, substitution with
heavier isotopes such as deuterium affords greater metabolic
stability (for example, increased in vivo half-life or reduced
dosage requirements). In yet other embodiments, substitution with
positron emitting isotopes, such as .sup.11C, .sup.18F, .sup.15O
and .sup.13N, is useful in Positron Emission Topography (PET)
studies for examining substrate receptor occupancy.
Isotopically-labeled compounds are prepared by any suitable method
or by processes using an appropriate isotopically-labeled reagent
in place of the non-labeled reagent otherwise employed.
[0105] In certain embodiments, the compounds described herein are
labeled by other means, including, but not limited to, the use of
chromophores or fluorescent moieties, bioluminescent labels, or
chemiluminescent labels.
[0106] Compounds of the invention can in certain embodiments form
acids or bases. In certain embodiments, the invention contemplates
acid addition salts. In other embodiments, the invention
contemplates base addition salts. In yet other embodiments, the
invention contemplates pharmaceutically acceptable acid addition
salts. In yet other embodiments, the invention contemplates
pharmaceutically acceptable base addition salts. Pharmaceutically
acceptable salts refer to salts of those bases or acids that are
not toxic or otherwise biologically undesirable.
[0107] Suitable pharmaceutically acceptable acid addition salts may
be prepared from an inorganic acid or from an organic acid.
Examples of inorganic acids include hydrochloric, hydrobromic,
hydriodic, nitric, carbonic, sulfuric (including sulfate and
hydrogen sulfate), and phosphoric acids (including hydrogen
phosphate and dihydrogen phosphate). Appropriate organic acids may
be selected from aliphatic, cycloaliphatic, aromatic, araliphatic,
heterocyclic, carboxylic and sulfonic classes of organic acids,
examples of which include formic, acetic, propionic, succinic,
glycolic, gluconic, lactic, malic, tartaric, citric, ascorbic,
glucuronic, maleic, malonic, saccharin, fumaric, pyruvic, aspartic,
glutamic, benzoic, anthranilic, 4-hydroxybenzoic, phenylacetic,
mandelic, embonic (pamoic), methanesulfonic, ethanesulfonic,
benzenesulfonic, pantothenic, trifluoromethanesulfonic,
2-hydroxyethanesulfonic, p-toluenesulfonic, sulfanilic,
cyclohexylaminosulfonic, stearic, alginic, .beta.-hydroxybutyric,
salicylic, galactaric and galacturonic acid.
[0108] Suitable pharmaceutically acceptable base addition salts of
compounds of the invention include, for example, metallic salts
including alkali metal, alkaline earth metal and transition metal
salts such as, for example, calcium, magnesium, potassium, sodium,
lithium and copper, iron and zinc salts. Pharmaceutically
acceptable base addition salts also include organic salts made from
basic amines such as, for example, N,N'-dibenzylethylene-diamine,
chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine
(N-methylglucamine) and procaine. All of these salts may be
prepared from the corresponding compound by reacting, for example,
the appropriate acid or base with the compound.
[0109] Disclosed is a kit comprising any composition of the
invention and an instructional material which describes
administering the composition to a tissue of a subject, such as a
mammal. This kit may comprise a (preferably sterile) solvent
suitable for dissolving or suspending the composition of the
invention prior to administering the composition to the subject,
such as a mammal.
Methods
[0110] The invention provides methods of preparing the lipid-based
nanoparticle of the invention. In certain embodiments, the method
comprises contacting in an aqueous system cholesterol, dicetyl
phosphate, amphipathic lipid, and hepatocyte receptor binding
molecule. In other embodiments, the method comprises contacting in
an aqueous system cholesterol, dicetyl phosphate, amphipathic
lipid, hepatocyte receptor binding molecule, and at least one
compound selected from the group consisting of a stabilizer and
stearoyl lysophosphatidylcholine. In yet other embodiments, the
method comprises contacting in an aqueous system cholesterol,
dicetyl phosphate, DSPC, and biotin-DHPE. In yet other embodiments,
the method comprises contacting in an aqueous system cholesterol,
dicetyl phosphate, DSPC, stearoyl lysophosphatidylcholine,
m-cresol, and biotin-DHPE.
[0111] In certain embodiments, the nanoparticle is formed in the
absence of the therapeutic agent, wherein optionally the
nanoparticle is at least partially concentrated, purified or
isolated, and wherein the therapeutic agent is contacted with the
nanoparticle, whereby at least a portion of the therapeutic agent
is dispersed within the nanoparticle.
[0112] In certain embodiments, the composition is treated with
cellulose acetate phthalate, which can bind non-covalently to at
least a portion of the therapeutic agent dispersed within the
nanoparticle and protect the therapeutic agent from metabolic
degradation. In other embodiments, the cellulose acetate phthalate
is covalently bound to the therapeutic agent and/or any of the
lipids that constitute the nanoparticle.
[0113] Further embodiments relating to certain methods for
preparing and/or processing and/or purifying a nanoparticle can be
found, for example, in U.S. Patent Application Nos. US20110135725
and US20090087479 and PCT Patent Application Publication No. WO
2018/169954, all of which are incorporated herein in their
entireties by reference.
[0114] The invention further provides a method of treating a
disease in a mammal. In certain embodiments, the method comprises
administering to the mammal in need thereof a therapeutically
effective amount of a nanoparticle and/or a composition of the
invention.
[0115] In certain embodiments, the disease is diabetes mellitus and
the therapeutic agent comprises insulin. In other embodiments, the
therapeutic agent further comprises a GLP-1 agonist and/or
serotonin.
Administration/Dosage/Formulations
[0116] The invention also encompasses pharmaceutical compositions
and methods of their use. These pharmaceutical compositions may
comprise an active ingredient (which can be one or more
compositions of the invention, or pharmaceutically acceptable salts
thereof) optionally in combination with one or more
pharmaceutically acceptable agents. The compositions set forth
herein can be used alone or in combination with additional
compounds to produce additive, complementary, or synergistic
effects.
[0117] The regimen of administration may affect what constitutes an
effective amount. The therapeutic formulations may be administered
to the subject either prior to or after the onset of a disease or
disorder contemplated herein. Further, several divided dosages, as
well as staggered dosages may be administered daily or
sequentially, or the dose may be continuously infused, or may be a
bolus injection, or may be administered inhalationally, buccally
and/or orally. Further, the dosages of the therapeutic formulations
may be proportionally increased or decreased as indicated by the
exigencies of the therapeutic or prophylactic situation.
[0118] Administration of the compositions of the present invention
to a patient, preferably a mammal, more preferably a human, may be
carried out using known procedures, at dosages and for periods of
time effective to treat a disease or disorder contemplated herein.
An effective amount of the therapeutic compound necessary to
achieve a therapeutic effect may vary according to factors such as
the state of the disease or disorder in the patient; the age, sex,
and weight of the patient; and the ability of the therapeutic
compound to treat a disease or disorder contemplated herein. Dosage
regimens may be adjusted to provide the optimum therapeutic
response. For example, several divided doses may be administered
daily or the dose may be proportionally reduced as indicated by the
exigencies of the therapeutic situation. A non-limiting example of
an effective dose range for a therapeutic compound of the invention
is from about 1 and 5,000 mg/kg of body weight/per day. One of
ordinary skill in the art would be able to study the relevant
factors and make the determination regarding the effective amount
of the therapeutic compound without undue experimentation.
[0119] Actual dosage levels of the active ingredients in the
pharmaceutical compositions of this invention may be varied so as
to obtain an amount of the active ingredient that is effective to
achieve the desired therapeutic response for a particular patient,
composition, and mode of administration, without being toxic to the
patient.
[0120] In particular, the selected dosage level depends upon a
variety of factors including the activity of the particular
compound employed, the time of administration, the rate of
excretion of the compound, the duration of the treatment, other
drugs, compounds or materials used in combination with the
compound, the age, sex, weight, condition, general health and prior
medical history of the patient being treated, and like factors
well, known in the medical arts.
[0121] A medical doctor, e.g., physician or veterinarian, having
ordinary skill in the art may readily determine and prescribe the
effective amount of the pharmaceutical composition required. For
example, the physician or veterinarian could start doses of the
compounds of the invention employed in the pharmaceutical
composition at levels lower than that required in order to achieve
the desired therapeutic effect, and gradually increase the dosage
until the desired effect is achieved.
[0122] In particular embodiments, it is especially advantageous to
formulate the compound in dosage unit form for ease of
administration and uniformity of dosage. Dosage unit form as used
herein refers to physically discrete units suited as unitary
dosages for the patients to be treated; each unit containing a
predetermined quantity of therapeutic compound calculated to
produce the desired therapeutic effect in association with the
required pharmaceutical vehicle. The dosage unit forms of the
invention are dictated by and directly dependent on (a) the unique
characteristics of the therapeutic compound and the particular
therapeutic effect to be achieved, and (b) the limitations inherent
in the art of compounding/formulating such a therapeutic compound
for the treatment of a disease or disorder contemplated herein.
[0123] In certain embodiments, the compositions of the invention
are formulated using one or more pharmaceutically acceptable
excipients or carriers. In certain embodiments, the pharmaceutical
compositions of the invention comprise a therapeutically effective
amount of a compound of the invention and a pharmaceutically
acceptable carrier.
[0124] The carrier may be a solvent or dispersion medium
containing, for example, water, ethanol, polyol (for example,
glycerol, propylene glycol, and liquid polyethylene glycol, and the
like), suitable mixtures thereof, and vegetable oils, as long as
the solvent or dispersion medium does not disrupt the nanoparticle
significantly. Prevention of the action of microorganisms may be
achieved by various antibacterial and antifungal agents, for
example, parabens, chlorobutanol, phenol, ascorbic acid,
thimerosal, and the like. In many cases, it is preferable to
include isotonic agents, for example, sugars, sodium chloride, or
polyalcohols such as mannitol and sorbitol, in the composition.
Prolonged absorption of the injectable compositions may be brought
about by including in the composition an agent that delays
absorption, for example, aluminum monostearate or gelatin.
[0125] In certain embodiments, the compositions of the invention
are administered to the patient in dosages that range from one to
five times per day or more. In other embodiments, the compositions
of the invention are administered to the patient in range of
dosages that include, but are not limited to, once every day, every
two, days, every three days to once a week, and once every two
weeks. It is readily apparent to one skilled in the art that the
frequency of administration of the various combination compositions
of the invention varies from individual to individual depending on
many factors including, but not limited to, age, disease or
disorder to be treated, gender, overall health, and other factors.
Thus, the invention should not be construed to be limited to any
particular dosage regime and the precise dosage and composition to
be administered to any patient is determined by the attending
physical taking all other factors about the patient into
account.
[0126] Compounds of the invention for administration may be in the
range of from about 1 .mu.g to about 10,000 mg, about 20 .mu.g to
about 9,500 mg, about 40 .mu.g to about 9,000 mg, about 75 .mu.g to
about 8,500 mg, about 150 .mu.g to about 7,500 mg, about 200 .mu.g
to about 7,000 mg, about 350 .mu.g to about 6,000 mg, about 500
.mu.g to about 5,000 mg, about 750 .mu.g to about 4,000 mg, about 1
mg to about 3,000 mg, about 10 mg to about 2,500 mg, about 20 mg to
about 2,000 mg, about 25 mg to about 1,500 mg, about 30 mg to about
1,000 mg, about 40 mg to about 900 mg, about 50 mg to about 800 mg,
about 60 mg to about 750 mg, about 70 mg to about 600 mg, about 80
mg to about 500 mg, and any and all whole or partial increments
there between.
[0127] In certain embodiments, the dose of a compound and/or
composition of the invention is from about 1 mg and about 2,500 mg.
In other embodiments, a dose of a compound of the invention used in
compositions described herein is less than about 10,000 mg, or less
than about 8,000 mg, or less than about 6,000 mg, or less than
about 5,000 mg, or less than about 3,000 mg, or less than about
2,000 mg, or less than about 1,000 mg, or less than about 500 mg,
or less than about 200 mg, or less than about 50 mg. Similarly, in
other embodiments, a dose of a second compound as described herein
is less than about 1,000 mg, or less than about 800 mg, or less
than about 600 mg, or less than about 500 mg, or less than about
400 mg, or less than about 300 mg, or less than about 200 mg, or
less than about 100 mg, or less than about 50 mg, or less than
about 40 mg, or less than about 30 mg, or less than about 25 mg, or
less than about 20 mg, or less than about 15 mg, or less than about
10 mg, or less than about 5 mg, or less than about 2 mg, or less
than about 1 mg, or less than about 0.5 mg, and any and all whole
or partial increments thereof.
[0128] In certain embodiments, the present invention is directed to
a packaged pharmaceutical composition comprising a container
holding a therapeutically effective amount of a compound and/or
composition of the invention, alone or in combination with a second
pharmaceutical agent; and instructions for using the compound to
treat, prevent, or reduce one or more symptoms of a disease or
disorder contemplated herein.
[0129] In certain embodiments, the container holds a lipid-based
nanoparticle, which does not comprise a therapeutic agent of
interest, such as but not limited to an insulin or a derivative or
analog thereof. In other embodiments, the container holds a
lipid-based nanoparticle, which comprises a therapeutic agent of
interest, such as but not limited to an insulin or a derivative or
analog thereof. In yet other embodiments, the container further
holds a therapeutic agent of interest, such as but not limited to
an insulin or a derivative or analog thereof.
Illustrative Non-Limiting Methods of Treating
[0130] Patients with Type 1 or Type 2 diabetes mellitus, as well as
patients with metabolic derangements, such as but not limited to
metabolic syndrome with elevated insulin levels, steatosis, and/or
steatohepatitis, can be administered an effective amount of a
nanoparticle of the invention comprising an insulin. When this
composition is administered subcutaneously, a portion of the
composition enters the circulatory system where the composition is
transported to the liver and other areas. The extended amphipathic
lipid binds the lipid construct to receptors of hepatocytes. A
portion of the administered composition is exposed to an external
gradient in vivo, where insulin can be solubilized and then move
from the lipid construct thereby supplying insulin to the muscle
and adipose tissue. Insulin that remains with the lipid construct
maintains the capability of being directed to the hepatocyte
binding receptor on the hepatocytes in the liver. Therefore, two
forms of insulin are produced from this particular lipid construct.
In an in vivo setting, free and lipid associated insulin are
generated in a time-dependent manner.
[0131] Administration of the nanoparticles and compositions
comprising same can be through any of the accepted modes of
administration for insulin that are desired to be administered.
These methods include oral, parenteral, nasal and other systemic or
aerosol forms. These methods further include pump delivery
systems.
[0132] Oral administration of a nanoparticle of the invention is
followed by intestinal absorption of insulin associated with the
nanoparticle of the invention into the circulatory system of the
body, where it is also exposed to the physiological pH of the
blood. The nanoparticle is targeted for delivery to the liver and
may be shielded by the presence of cellulose acetate phthalate
within the nanoparticle of the invention. In the case of oral
administration, the shielded nanoparticle transverses the oral
cavity, migrates through the stomach and moves into the small
intestine, where the alkaline pH of the small intestine degrades
the cellulose acetate phthalate shield. The deshielded nanoparticle
is absorbed into the circulatory system. This enables the
nanoparticle to be delivered to the sinusoids of the liver. A
receptor binding molecule, such as
1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-(Cap Biotinyl)
or any other hepatocyte specific molecule, provides a means for
lipid construct to bind to the receptor and then be engulfed or
endocytosed by the hepatocytes. Insulin is then released from the
nanoparticle where, upon gaining access to the cellular
environment, it performs its designated function with regard to
acting as an agent to control diabetes mellitus.
[0133] Patients with Type 1 or Type 2 diabetes mellitus, as well as
patients with metabolic derangements, such as but not limited to
metabolic syndrome with elevated insulin levels, steatosis, and/or
steatohepatitis, may be administered an effective amount of a
nanoparticle comprising a mixture of free glargine insulin and
glargine insulin associated with the nanoparticle. Glargine insulin
can be combined with other forms of insulin, such as insulin
lispro, insulin aspart (including FIASP.RTM., Novo Nordisk),
regular insulin, insulin glargine, insulin zinc, extended human
insulin zinc suspension, isophane insulin, human buffered regular
insulin, insulin glulisine, recombinant human regular insulin,
recombinant human insulin isophane, insulin detemir, biphasic human
insulin, and insulin degludec (including TRESIBA.RTM., Novo
Nordisk) or premixed combinations of any of the aforementioned
insulins, a derivative thereof, and a combination of any of the
aforementioned insulins. The composition can be administered by a
subcutaneous or oral route.
[0134] After a composition is administered to a patient by
subcutaneous injection, the in situ physiological environment in
the injection area, the morphology and chemical structures of free
insulin and the insulin associated with the nanoparticle begin to
change. For example, as the pH of the environment around the free
glargine insulin and the glargine insulin associated with the
nanoparticle increases after being diluted with physiological
media, the pH reaches the isoelectric point of glargine insulin,
where flocculation, aggregation and precipitation reactions occur
for both free glargine insulin and glargine insulin associated with
the nanoparticle. In certain embodiments, free glargine insulin
changes from a soluble form at injection, to a insoluble form at a
pH near its isoelectric point of pH 5.8-6.2, and then to a soluble
form at physiological pH. The rates at which these processes occur
differ between free glargine insulin and glargine insulin
associated with the nanoparticle. The free glargine insulin is
directly exposed to changes in pH and dilution. Exposure of
glargine insulin associated with the nanoparticle to small changes
in pH and dilution at physiological pH is delayed due to the time
required for diffusion of physiological fluids or media through the
lipid bilayer in the nanoparticle. The delay in the release of
insulin from the lipid construct as well as the delay of the
release of the insulin associated with the nanoparticle is a
feature of the invention since it affects and augments the
biological and pharmacological response in vivo.
[0135] Oral administration of a pharmaceutical composition that
combines free glargine insulin and glargine insulin associated with
a nanoparticle is followed by intestinal absorption of glargine
insulin associated with the nanoparticle into the circulatory
system of the body, where it is also exposed to the physiological
pH of the blood. In certain embodiments, the composition comprises
a delayed release matrix which releases HDV glargine over a
prolonged period of time, in order to achieve a 24-hour dose
regimen. All or a portion of the nanoparticle is delivered to the
liver.
[0136] Patients with Type 1 or Type 2 diabetes mellitus, as well as
patients with metabolic derangements, such as but not limited to
metabolic syndrome with elevated insulin levels, steatosis, and/or
steatohepatitis, can be administered an effective amount of a
hepatocyte targeted composition comprising a mixture of free
recombinant human insulin isophane (NPH) plus free recombinant
human regular insulin along with recombinant human insulin isophane
and recombinant human regular insulin which are both associated
with a nanoparticle. Recombinant human insulin isophane can be
combined with other forms of insulin, such insulin lispro, insulin
aspart (including FIASP.RTM., Novo Nordisk), regular insulin,
insulin glargine, insulin zinc, extended human insulin zinc
suspension, isophane insulin, human buffered regular insulin,
insulin glulisine, recombinant human regular insulin, recombinant
human insulin isophane, insulin detemir, biphasic human insulin,
and insulin degludec (including TRESIBA.RTM., Novo Nordisk,
ultralong-acting basal insulin analogue; has one single amino acid
deleted in comparison to human insulin, and is conjugated to
hexadecanedioic acid via gamma-L-glutamyl spacer at the amino acid
lysine at position B29), or any (premixed) combinations
thereof.
[0137] In certain embodiments, the composition comprises a delayed
release matrix which releases HDV NPH over a prolonged period of
time, in order to achieve a 24-hour dose regimen.
[0138] Oral administration of a pharmaceutical composition that
combines free recombinant human insulin isophane and recombinant
human insulin isophane associated with a nanoparticle is followed
by intestinal absorption of recombinant human insulin isophane
associated with the nanoparticle into the circulatory system of the
body where it is also exposed to the physiological pH of the blood.
All or a portion of the nanoparticle is delivered to the liver,
while the non-HDV isophane is slowly absorbed from a slow release
matrix for release into the general circulation.
[0139] As the physiological dilution is increased in situ in the
subcutaneous space or upon entering into the circulatory system,
free recombinant human insulin isophane and recombinant human
insulin isophane associated with the nanoparticle encounter a
normal physiological pH environment of pH 7.4. As a result of
dilution free recombinant human insulin isophane changes from an
insoluble form at injection, to a soluble form at physiological pH.
In the soluble form, recombinant human insulin isophane migrates
through the body to sites where it is capable of eliciting a
pharmacological response. Recombinant human insulin isophane
associated with the nanoparticle becomes solubilized and released
from the nanoparticle at a different rate that is slower than that
of free recombinant human insulin isophane. This is because
recombinant human insulin isophane associated with the nanoparticle
has to traverse the core volume and lipid domains of the
nanoparticle before it contacts the bulk phase media.
[0140] The amount of insulin administered will be dependent on the
subject being treated, the type and severity of the affliction, the
manner of administration and the judgment of the prescribing
physician. Although effective dosage ranges for specific
biologically active substances of interest are dependent upon a
variety of factors and are generally known to one of ordinary skill
in the art, some dosage guidelines can be generally defined. For
most forms of administration, the nanoparticle will be suspended in
an aqueous solution and generally not exceed 4.0% (w/v) of the
total formulation. The drug component of the formulation will in
certain embodiments be less than 20% (w/v) of the formulation and
generally greater than 0.01% (w/v).
[0141] In certain embodiments, the pharmaceutical composition
comprises HDV insulin, and no free insulin. In such cases, all of
the insulin within the composition is targeted to the liver. In
other embodiments, the pharmaceutical composition comprises HDV
insulin and free insulin (non-HDV insulin). The ratio between HDV
insulin and free insulin can be, in non-limiting example, about
0.1:99.9, 0.2:99.8, 0.3:99.7, 0.4:99.6, 0.5:99.5, 0.6:99.4,
0.7:99.3, 0.8:99.2, 0.9:99.1, 1:99, 2:98, 3:97, 4:96, 5:95, 6:94,
7:93, 8:92, 9:91, 10:90, 12:88, 14:86, 16:84, 18:82, 20:80, 22:78,
24:76, 25:75, 26:74, 28:72, 30:70, 32:68, 34:66, 36:64, 38:62,
40:60, 42:58, 44:56, 46:54, 48:52, and/or 50:50.
[0142] Dosage forms or compositions containing active ingredient in
the range of 0.005% to 5% with the balance made up from non-toxic
carriers can be prepared.
[0143] The exact composition of these formulations may vary widely
depending on the particular properties of the drug in question. In
certain embodiments, they comprise from 0.01% to 5%, and preferably
from 0.05% to 1% active ingredient for highly potent drugs, and
from 2%-4% for moderately active drugs.
[0144] The percentage of active ingredient contained in such
parenteral compositions is highly dependent on the specific nature
thereof, as well as the activity of the active ingredient and the
needs of the subject. However, percentages of active ingredient of
0.01% to 5% in solution are employable, and will be higher if the
composition is a solid which will be subsequently diluted to the
above percentages. In certain embodiments, the composition
comprises 0.2%-2.0% of the active agent in solution.
Administration
[0145] Formulations may be employed in admixtures with conventional
excipients, i.e., pharmaceutically acceptable organic or inorganic
carrier substances suitable for oral, parenteral, nasal,
intravenous, subcutaneous, enteral, or any other suitable mode of
administration, known to the art. The pharmaceutical preparations
may be sterilized and if desired mixed with auxiliary agents, e.g.,
lubricants, preservatives, stabilizers, wetting agents,
emulsifiers, salts for influencing osmotic pressure buffers,
coloring, flavoring and/or aromatic substances and the like. They
may also be combined where desired with other active agents, e.g.,
other analgesic agents.
[0146] Routes of administration of any of the compositions of the
invention include oral, nasal, rectal, intravaginal, parenteral,
buccal, sublingual or topical. The compounds and/or compositions
for use in the invention may be formulated for administration by
any suitable route, such as for oral or parenteral, for example,
transdermal, transmucosal (e.g., sublingual, lingual,
(trans)buccal, (trans)urethral, vaginal (e.g., trans- and
perivaginally), (intra)nasal and (trans)rectal), intravesical,
intrapulmonary, intraduodenal, intragastrical, intrathecal,
subcutaneous, intramuscular, intradermal, intra-arterial,
intravenous, intrabronchial, inhalation, and topical
administration.
[0147] Suitable compositions and dosage forms include, for example,
tablets, capsules, caplets, pills, gel caps, troches, dispersions,
suspensions, solutions, syrups, granules, beads, transdermal
patches, gels, powders, pellets, magmas, lozenges, creams, pastes,
plasters, lotions, discs, suppositories, liquid sprays for nasal or
oral administration, dry powder or aerosolized formulations for
inhalation, compositions and formulations for intravesical
administration and the like. It should be understood that the
formulations and compositions that would be useful in the present
invention are not limited to the particular formulations and
compositions that are described herein.
[0148] Oral Administration
[0149] For oral application, particularly suitable are tablets,
dragees, liquids, drops, suppositories, or capsules, caplets and
gelcaps. The compositions intended for oral use may be prepared
according to any method known in the art and such compositions may
contain one or more agents selected from the group consisting of
inert, non-toxic pharmaceutically excipients that are suitable for
the manufacture of tablets. Such excipients include, for example an
inert diluent such as lactose; granulating and disintegrating
agents such as cornstarch; binding agents such as starch; and
lubricating agents such as magnesium stearate. The tablets may be
uncoated or they may be coated by known techniques for elegance or
to delay the release of the active ingredients. Formulations for
oral use may also be presented as hard gelatin capsules wherein the
active ingredient is mixed with an inert diluent.
[0150] For oral administration, the compounds and/or compositions
of the invention may be in the form of tablets or capsules prepared
by conventional means with pharmaceutically acceptable excipients
such as binding agents (e.g., polyvinylpyrrolidone,
hydroxypropylcellulose or hydroxypropyl methylcellulose); fillers
(e.g., cornstarch, lactose, microcrystalline cellulose or calcium
phosphate); lubricants (e.g., magnesium stearate, talc, or silica);
disintegrates (e.g., sodium starch glycollate); or wetting agents
(e.g., sodium lauryl sulphate). If desired, the tablets may be
coated using suitable methods and coating materials such as
OPADRY.TM. film coating systems available from Colorcon, West
Point, Pa. (e.g., OPADRY.TM. OY Type, OYC Type, Organic Enteric
OY-P Type, Aqueous Enteric OY-A Type, OY-PM Type and OPADRY.TM.
White, 32K18400). Liquid preparation for oral administration may be
in the form of solutions, syrups or suspensions. The liquid
preparations may be prepared by conventional means with
pharmaceutically acceptable additives such as suspending agents
(e.g., sorbitol syrup, methyl cellulose or hydrogenated edible
fats); emulsifying agent (e.g., lecithin or acacia); non-aqueous
vehicles (e.g., almond oil, oily esters or ethyl alcohol); and
preservatives (e.g., methyl or propyl p-hydroxy benzoates or sorbic
acid).
[0151] Parenteral Administration
[0152] For parenteral administration, the compounds and/or
compositions of the invention may be formulated for injection or
infusion, for example, intravenous, intramuscular or subcutaneous
injection or infusion, or for administration in a bolus dose and/or
continuous infusion. Suspensions, solutions or emulsions in an oily
or aqueous vehicle, optionally containing other formulatory agents
such as suspending, stabilizing and/or dispersing agents may be
used.
[0153] Pulmonary Administration
[0154] A pharmaceutical composition of the invention may be
prepared, packaged, or sold in a formulation suitable for pulmonary
administration via the buccal cavity. Such a formulation may
comprise dry particles which comprise the active ingredient and
which have a diameter in the range from about 0.5 to about 7
microns, and preferably from about 1 to about 6 microns. Such
compositions are conveniently in the form of dry powders for
administration using a device comprising a dry powder reservoir to
which a stream of propellant may be directed to disperse the powder
or using a self-propelling solvent/powder-dispensing container such
as a device comprising the active ingredient dissolved or suspended
in a low-boiling propellant in a sealed container. Preferably, such
powders comprise particles wherein at least 98% of the particles by
weight have a diameter greater than 0.5 microns and at least 95% of
the particles by number have a diameter less than 7 microns. More
preferably, at least 95% of the particles by weight have a diameter
greater than 1 nanometer and at least 90% of the particles by
number have a diameter less than 6 microns. Dry powder compositions
preferably include a solid fine powder diluent such as sugar and
are conveniently provided in a unit dose form.
[0155] Low boiling propellants generally include liquid propellants
having a boiling point of below 65.degree. F. at atmospheric
pressure. Generally, the propellant may constitute 50 to 99.9%
(w/w) of the composition, and the active ingredient may constitute
0.1 to 20% (w/w) of the composition. The propellant may further
comprise additional ingredients such as a liquid non-ionic or solid
anionic surfactant or a solid diluent (preferably having a particle
size of the same order as particles comprising the active
ingredient).
[0156] Pharmaceutical compositions of the invention formulated for
pulmonary delivery may also provide the active ingredient in the
form of droplets of a solution or suspension. Such formulations may
be prepared, packaged, or sold as aqueous or dilute alcoholic
solutions or suspensions, optionally sterile for administration by
injection, comprising the active ingredient, and may conveniently
be administered using any nebulization or atomization device. In
certain embodiments, the compounds and/or compositions of the
invention are sterile filtered before administration to the
subject. Such formulations may further comprise one or more
additional ingredients including, but not limited to, a flavoring
agent such as saccharin sodium, a volatile oil, a buffering agent,
a surface active agent, or a preservative such as
methylhydroxybenzoate. The droplets provided by this route of
administration preferably have an average diameter in the range
from about 0.1 to about 200 microns.
[0157] Intranasal Delivery
[0158] The formulations described herein as being useful for
pulmonary delivery are also useful for intranasal delivery of a
pharmaceutical composition of the invention.
[0159] Another formulation suitable for intranasal administration
is a coarse powder comprising the active ingredient and having an
average particle from about 0.2 to 500 microns. Such a formulation
is administered in the manner in which snuff is taken i.e. by rapid
inhalation through the nasal passage from a container of the powder
held close to the nares.
[0160] Formulations suitable for nasal administration may, for
example, comprise from about as little as 0.1% (w/w) and as much as
75% (w/w) of the active ingredient, and may further comprise one or
more of the additional ingredients described herein.
[0161] Additional Administration Forms
[0162] Additional dosage forms of this invention include dosage
forms as described in U.S. Pat. Nos. 6,340,475; 6,488,962;
6,451,808; 5,972,389; 5,582,837; and 5,007,790. Additional dosage
forms of this invention also include dosage forms as described in
U.S. Patent Applications Nos. 20030147952; 20030104062;
20030104053; 20030044466; 20030039688; and 20020051820. Additional
dosage forms of this invention also include dosage forms as
described in PCT Applications Nos. WO 03/35041; WO 03/35040; WO
03/35029; WO 03/35177; WO 03/35039; WO 02/96404; WO 02/32416; WO
01/97783; WO 01/56544; WO 01/32217; WO 98/55107; WO 98/11879; WO
97/47285; WO 93/18755; and WO 90/11757.
[0163] Controlled Release Formulations and Drug Delivery
Systems
[0164] In certain embodiments, the formulations of the present
invention may be, but are not limited to, short-term, rapid-offset,
as well as controlled, for example, sustained release, delayed
release and pulsatile release formulations.
[0165] The term sustained release is used in its conventional sense
to refer to a drug formulation that provides for gradual release of
a drug over an extended period of time, and that may, although not
necessarily, result in substantially constant blood levels of a
drug over an extended time period. The period of time may be as
long as a month or more and should be a release that is longer that
the same amount of agent administered in bolus form.
[0166] For sustained release, the compositions may be formulated
with a suitable polymer or hydrophobic material that provides
sustained release properties to the compounds and/or compositions.
As such, the compositions and/or compositions for use the method of
the invention may be administered in the form of microparticles,
for example, by injection or in the form of wafers or discs by
implantation.
[0167] In certain embodiments, the compounds and/or compositions of
the invention are administered to a patient, alone or in
combination with another pharmaceutical agent, using a sustained
release formulation.
[0168] The term delayed release is used herein in its conventional
sense to refer to a drug formulation that provides for an initial
release of the drug after some delay following drug administration
and that mat, although not necessarily, includes a delay of from
about 10 minutes up to about 12 hours.
[0169] The term pulsatile release is used herein in its
conventional sense to refer to a drug formulation that provides
release of the drug in such a way as to produce pulsed plasma
profiles of the drug after drug administration.
[0170] The term immediate release is used in its conventional sense
to refer to a drug formulation that provides for release of the
drug immediately after drug administration.
[0171] As used herein, short-term refers to any period of time up
to and including about 8 hours, about 7 hours, about 6 hours, about
5 hours, about 4 hours, about 3 hours, about 2 hours, about 1 hour,
about 40 minutes, about 20 minutes, or about 10 minutes and any or
all whole or partial increments thereof after drug administration
after drug administration.
[0172] As used herein, rapid-offset refers to any period of time up
to and including about 8 hours, about 7 hours, about 6 hours, about
5 hours, about 4 hours, about 3 hours, about 2 hours, about 1 hour,
about 40 minutes, about 20 minutes, or about 10 minutes, and any
and all whole or partial increments thereof after drug
administration.
Dosing
[0173] The therapeutically effective amount or dose of a compound
and/or composition of the present invention depends on the age, sex
and weight of the patient, the current medical condition of the
patient and the progression of a disease or disorder contemplated
herein in the patient being treated. The skilled artisan is able to
determine appropriate dosages depending on these and other
factors.
[0174] A suitable dose of a compound and/or composition of the
present invention may be in the range of from about 0.01 mg to
about 5,000 mg per day, such as from about 0.1 mg to about 1,000
mg, for example, from about 1 mg to about 500 mg, such as about 5
mg to about 250 mg per day. The dose may be administered in a
single dosage or in multiple dosages, for example from 1 to 4 or
more times per day. When multiple dosages are used, the amount of
each dosage may be the same or different. For example, a dose of 1
mg per day may be administered as two 0.5 mg doses, with about a
12-hour interval between doses.
[0175] It is understood that the amount of compound and/or
composition dosed per day may be administered, in non-limiting
examples, every day, every other day, every 2 days, every 3 days,
every 4 days, or every 5 days. For example, with every other day
administration, a 5 mg per day dose may be initiated on Monday with
a first subsequent 5 mg per day dose administered on Wednesday, a
second subsequent 5 mg per day dose administered on Friday, and so
on.
[0176] In the case wherein the patient's status does improve, upon
the doctor's discretion the administration of the inhibitor of the
invention is optionally given continuously; alternatively, the dose
of drug being administered is temporarily reduced or temporarily
suspended for a certain length of time (i.e., a "drug holiday").
The length of the drug holiday optionally varies between 2 days and
1 year, including by way of example only, 2 days, 3 days, 4 days, 5
days, 6 days, 7 days, 10 days, 12 days, 15 days, 20 days, 28 days,
35 days, 50 days, 70 days, 100 days, 120 days, 150 days, 180 days,
200 days, 250 days, 280 days, 300 days, 320 days, 350 days, or 365
days. The dose reduction during a drug holiday includes from
10%-100%, including, by way of example only, 10%, 15%, 20%, 25%,
30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%,
95%, or 100%.
[0177] Once improvement of the patient's conditions has occurred, a
maintenance dose is administered if necessary. Subsequently, the
dosage or the frequency of administration, or both, is reduced, as
a function of the viral load, to a level at which the improved
disease is retained. In certain embodiments, patients require
intermittent treatment on a long-term basis upon any recurrence of
symptoms and/or infection.
[0178] The compounds and/or compositions for use in the method of
the invention may be formulated in unit dosage form. The term "unit
dosage form" refers to physically discrete units suitable as
unitary dosage for patients undergoing treatment, with each unit
containing a predetermined quantity of active material calculated
to produce the desired therapeutic effect, optionally in
association with a suitable pharmaceutical carrier. The unit dosage
form may be for a single daily dose or one of multiple daily doses
(e.g., about 1 to 4 or more times per day). When multiple daily
doses are used, the unit dosage form may be the same or different
for each dose.
[0179] Toxicity and therapeutic efficacy of such therapeutic
regimens are optionally determined in cell cultures or experimental
animals, including, but not limited to, the determination of the
LD.sub.50 (the dose lethal to 50% of the population) and the
ED.sub.50 (the dose therapeutically effective in 50% of the
population). The dose ratio between the toxic and therapeutic
effects is the therapeutic index, which is expressed as the ratio
between LD.sub.50 and ED.sub.50. The data obtained from cell
culture assays and animal studies are optionally used in
formulating a range of dosage for use in human. The dosage of such
compounds and/or compositions lies preferably within a range of
circulating concentrations that include the ED.sub.50 with minimal
toxicity. The dosage optionally varies within this range depending
upon the dosage form employed and the route of administration
utilized.
Definitions
[0180] Unless defined otherwise, all technical and scientific terms
used herein generally have the same meaning as commonly understood
by one of ordinary skill in the art to which the invention belongs.
Generally, the nomenclature used herein and the laboratory
procedures in organic chemistry and protein chemistry are those
well known and commonly employed in the art.
[0181] The articles "a" and "an" are used herein to refer to one or
to more than one (i.e., to at least one) of the grammatical object
of the article. By way of example, "an element" means one element
or more than one element.
[0182] The term "A1c" or "A1c" or "HbA1c" or "hemoglobin A1c" or
"HBA1c" or "HgbA1c" or "haemoglobin A1c" or "HbA1c" or "Hb1c"
refers to a form of hemoglobin that is covalently bound to glucose.
A1c is formed in a non-enzymatic glycation pathway by hemoglobin's
exposure to plasma glucose. A1c is measured primarily to identify
the three-month average plasma glucose concentration, and thus can
be used as a diagnostic test for diabetes and as assessment test
for glycemic control in people with diabetes. The ratio of A1c to
total hemoglobin (% A1c) (generally measured as mass/mass) is used
to diagnose diabetes (according to 1993 Diabetes Control and
Complications Trial or DCCT): normal individuals have less than
5.7% A1, pre-diabetic individuals have 5-7-6.4% A1c, and diabetic
individuals have greater than 6.5% A1c. The DCCT % A1c value can be
converted to the International Federation of Clinical Chemistry and
Laboratory Medicine (IFCC) units using the formula:
IFCC HbA1c(mmol/mol)=[DCCT HbA1c(%)-2.14].times.10.929
[0183] As used herein, the term "about" is understood by persons of
ordinary skill in the art and varies to some extent on the context
in which it is used. As used herein when referring to a measurable
value such as an amount, a temporal duration, and the like, the
term "about" is meant to encompass variations of .+-.20% or
.+-.10%, more preferably .+-.5%, even more preferably .+-.1%, and
still more preferably .+-.0.1% from the specified value, as such
variations are appropriate to perform the disclosed methods.
[0184] As used herein, the term "active ingredient" refers to a
therapeutic agent that is to be delivered to a subject to produce a
therapeutic effect in the subject. Non-limiting examples of active
ingredients contemplated within the invention are insulin,
interferon, parathyroid hormone, calcitonin, serotonin, serotonin
agonist, serotonin reuptake inhibitor, human growth hormone, GIP,
anti-GIP monoclonal antibody, metformin, bromocriptine, dopamine,
glucagon and/or GLP-1.
[0185] The term "amphipathic lipid" means a lipid molecule having a
polar and non-polar end.
[0186] By "aqueous media" is meant water or water containing buffer
or salt.
[0187] As used herein, the term "basal insulin" or "background
insulin" is insulin that is taken to keep blood glucose levels at
consistent levels during periods of fasting. Basal insulin is thus
needed to keep blood glucose levels under control, and to allow the
cells to take in glucose for energy. Basal insulin is usually taken
once or twice a day depending on the insulin. Basal insulin needs
to act over a relatively long period of time, and thus is either
long acting insulin or intermediate insulin.
[0188] As used herein, the term "basal glucose control" refers to
the glucose control that is afforded by use of basal insulin, or an
equivalent thereof.
[0189] The term "bioavailability" refers to a measurement of the
rate and extent that insulin reaches the systemic circulation and
is available at the sites of action.
[0190] As used herein, the term "bolus insulin" refers to insulin
that is specifically taken just before, at, or just after meal
times to keep blood glucose levels under control following a meal.
Bolus insulin needs to act quickly and is generally short acting
insulin or rapid acting insulin.
[0191] As used herein, the term "bolus glucose control" refers to
the glucose control that is afforded by use of bolus insulin, or an
equivalent thereof.
[0192] As used herein, the term "CGM" refers to continuous glucose
monitoring.
[0193] In one aspect, the terms "co-administered" and
"co-administration" as relating to a subject to refer to
administering to the subject a compound of the invention or salt
thereof along with a compound that may also treat any disease or
disorder contemplated herein and/or with a compound that is useful
in treating other medical conditions but which in themselves may
cause or facilitate any disease or disorder contemplated herein. In
certain embodiments, the co-administered compounds are administered
separately, or in any kind of combination as part of a single
therapeutic approach. The co-administered compound may be
formulated in any kind of combinations as mixtures of solids and
liquids under a variety of solid, gel, and liquid formulations, and
as a solution.
[0194] As used herein, a "disease" is a state of health of a
subject wherein the subject cannot maintain homeostasis, and
wherein if the disease is not ameliorated then the subject's health
continues to deteriorate.
[0195] As used herein, a "disorder" in a subject is a state of
health in which the subject is able to maintain homeostasis, but in
which the subject's state of health is less favorable than it would
be in the absence of the disorder. Left untreated, a disorder does
not necessarily cause a further decrease in the subject's state of
health.
[0196] As used herein, the term "ED.sub.50" refers to the effective
dose of a formulation that produces 50% of the maximal effect in
subjects that are administered that formulation.
[0197] As used herein, an "effective amount," "therapeutically
effective amount" or "pharmaceutically effective amount" of a
compound is that amount of compound that is sufficient to provide a
beneficial effect to the subject to which the compound is
administered.
[0198] The term "free active ingredient" or "free therapeutic
agent" refers to an active ingredient or therapeutic agent that is
not dispersed within the lipid particle (i.e., located within,
adsorbed on and/or bound to the lipid particle membrane).
[0199] The terms "glargine" and "glargine insulin" both refer to a
recombinant human insulin analog which differs from human insulin
in that the amino acid asparagine at position A21 is replaced by
glycine and two arginines are added to the C-terminus of the
B-chain. Chemically, it is
21.sup.A-Gly-30.sup.Ba-L-Arg-30.sup.Bb-L-Arg-human insulin and has
the empirical formula C.sub.267H.sub.404N.sub.72O.sub.78S.sub.6 and
a molecular weight of 6063.
[0200] As used herein, the term "hyperinsulinemia" refers to a
condition in which there are excess levels of insulin circulating
in the blood relative to the level of glucose. Hyperinsulinemia can
be an unwanted side effect of administration of exogenous insulin
to a diabetic patient (thus being a form of iatrogenic
hyperinsulinemia; see Cryer, 2008, Diabetes 57(12):3169-76,
McCrinson & Sherwin, 2010, Diabetes 59(10):2333-9; Wang, et
al., 2013, J. Diab. & Its Compl. 27(1):70-74; all of which are
incorporated herein in their entireties by reference). That
condition can trigger complications such as metabolic disease,
hypoglycemia, increased risk of polycystic ovary syndrome (PCOS),
increased synthesis of VLDL (hypertriglyceridemia), hypertension
(insulin increases sodium retention by the renal tubules), coronary
artery disease (CAD; increased insulin damages endothelial cells),
increased risk of cardiovascular disease, and/or weight gain and
lethargy.
[0201] As used herein, the term "hypoglycemic event" or
"hypoglycemia event" refers to an event wherein the subject's blood
sugar is lower than 70 mg/dL for a significant amount of time, such
as but not limited to 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, or
60 minutes. In certain embodiments, a hypoglycemic event is defined
as a series of CGM values less than about 54 mg/dL, separated by 20
min or more, with no intervening values of 54 mg/dL or more. In
certain embodiments, a hypoglycemic event is defined as over 15 min
of CGM values less than about 54 mg/dL.
[0202] "Instructional material," as that term is used herein,
includes a publication, a recording, a diagram, or any other medium
of expression that can be used to communicate the usefulness of the
composition and/or compound of the invention in a kit. The
instructional material of the kit may, for example, be affixed to a
container that contains the compound and/or composition of the
invention or be shipped together with a container that contains the
compound and/or composition. Alternatively, the instructional
material may be shipped separately from the container with the
intention that the recipient uses the instructional material and
the compound cooperatively. Delivery of the instructional material
may be, for example, by physical delivery of the publication or
other medium of expression communicating the usefulness of the kit,
or may alternatively be achieved by electronic transmission, for
example by means of a computer, such as by electronic mail, or
download from a website.
[0203] The term "insulin" refers to natural or recombinant forms of
insulin, and derivatives of the aforementioned insulins. Examples
of insulin include, but are not limited to insulin lispro (such as,
for example, ADMELOG.RTM., Sanofi), insulin aspart (such as, for
example, FIASP.RTM., Novo Nordisk), regular insulin, insulin
glargine (such as, for example, BASAGLAR.RTM., Lilly), insulin
zinc, human insulin zinc extended, isophane insulin, human buffered
regular insulin, insulin glulisine, recombinant human regular
insulin, recombinant human insulin isophane, insulin detemir,
biphasic human insulin, and insulin degludec (including
TRESIBA.RTM., Novo Nordisk, ultralong-acting basal insulin
analogue; has one single amino acid deleted in comparison to human
insulin, and is conjugated to hexadecanedioic acid via
gamma-L-glutamyl spacer at the amino acid lysine at position B29).
Also included are animal insulins, such as bovine or porcine
insulin.
[0204] As used herein, the term "iotrogenic" refers to any illness
caused by a medical examination or treatment.
[0205] The term "isoelectric point" refers to the pH at which the
concentrations of positive and negative charges on the protein are
equal and, as a result, the protein will express a net zero charge.
At the isoelectric point, a protein will exist almost entirely in
the form of a zwitterion, or hybrid between forms of the protein.
Proteins are least stable at their isoelectric points, and are more
easily coagulated or precipitated at this pH. However, proteins are
not denatured upon isoelectric precipitation since this process is
essentially reversible.
[0206] The term "lipid construct" refers to a lipid and/or
phospholipid particle in which individual lipid molecules interact
to create a bipolar lipid membrane that defines the boundaries of
the lipid construct.
[0207] As the term is used herein, "to modulate" or "modulation of
a biological or chemical process or state refers to the alteration
of the normal course of the biological or chemical process, or
changing the state of the biological or chemical process to a new
state that is different than the present state. For example,
modulation of the isoelectric point of a polypeptide may involve a
change that increases the isoelectric point of the polypeptide.
Alternatively, modulation of the isoelectric point of a polypeptide
may involve a change that decreases the isoelectric point of a
polypeptide.
[0208] As used herein, a "metabolic derangement" refers to a
metabolic disorder or disease relating to uncontrolled, elevated,
or fluctuating insulin levels, such as but not limited to metabolic
syndrome with elevated insulin levels, steatosis, and/or
steatohepatitis.
[0209] The term "non-glargine insulin" refers at all insulins,
either natural or recombinant that are not glargine insulin. The
term includes insulin-like moieties, including fragments of insulin
molecules, that have biological activity of insulins.
[0210] As used herein, the term "pharmaceutical composition" or
"composition" refers to a mixture of at least one compound useful
within the invention with a pharmaceutically acceptable carrier.
The pharmaceutical composition facilitates administration of the
compound to a subject.
[0211] As used herein, the term "pharmaceutically acceptable"
refers to a material, such as a carrier or diluent, which does not
abrogate the biological activity or properties of the compound
useful within the invention, and is relatively non-toxic, i.e., the
material may be administered to a subject without causing
undesirable biological effects or interacting in a deleterious
manner with any of the components of the composition in which it is
contained.
[0212] As used herein, the term "pharmaceutically acceptable
carrier" means a pharmaceutically acceptable material, composition
or carrier, such as a liquid or solid filler, stabilizer,
dispersing agent, suspending agent, diluent, excipient, thickening
agent, solvent or encapsulating material, involved in carrying or
transporting a compound useful within the invention within or to
the subject such that it may perform its intended function.
Typically, such constructs are carried or transported from one
organ, or portion of the body, to another organ, or portion of the
body. Each carrier must be "acceptable" in the sense of being
compatible with the other ingredients of the formulation, including
the compound useful within the invention, and not injurious to the
subject. Some examples of materials that may serve as
pharmaceutically acceptable carriers include: sugars, such as
lactose, glucose and sucrose; starches, such as corn starch and
potato starch; cellulose, and its derivatives, such as sodium
carboxymethyl cellulose, ethyl cellulose and cellulose acetate;
powdered tragacanth; malt; gelatin; talc; excipients, such as cocoa
butter and suppository waxes; oils, such as peanut oil, cottonseed
oil, safflower oil, sesame oil, olive oil, corn oil and soybean
oil; glycols, such as propylene glycol; polyols, such as glycerin,
sorbitol, mannitol and polyethylene glycol; esters, such as ethyl
oleate and ethyl laurate; agar; buffering agents, such as magnesium
hydroxide and aluminum hydroxide; surface active agents; alginic
acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl
alcohol; phosphate buffer solutions; and other non-toxic compatible
substances employed in pharmaceutical formulations. As used herein,
"pharmaceutically acceptable carrier" also includes any and all
coatings, antibacterial and antifungal agents, and absorption
delaying agents, and the like that are compatible with the activity
of the compound useful within the invention, and are
physiologically acceptable to the subject. Supplementary active
compounds may also be incorporated into the compositions. The
"pharmaceutically acceptable carrier" may further include a
pharmaceutically acceptable salt of the compound useful within the
invention. Other additional ingredients that may be included in the
pharmaceutical compositions used in the practice of the invention
are known in the art and described, for example in Remington's
Pharmaceutical Sciences (Genaro, Ed., Mack Publishing Co., 1985,
Easton, Pa.), which is incorporated herein by reference.
[0213] As used herein, the language "pharmaceutically acceptable
salt" refers to a salt of the administered compound prepared from
pharmaceutically acceptable non-toxic acids and bases, including
inorganic acids, inorganic bases, organic acids, inorganic bases,
solvates, hydrates, and clathrates thereof.
[0214] The term "prevent," "preventing" or "prevention," as used
herein, means avoiding or delaying the onset of symptoms associated
with a disease or condition in a subject that has not developed
such symptoms at the time the administering of an agent or compound
commences.
[0215] Disease, condition and disorder are used interchangeably
herein.
[0216] By the term "specifically bind" or "specifically binds," as
used herein, is meant that a first molecule preferentially binds to
a second molecule (e.g., a particular receptor or enzyme), but does
not necessarily bind only to that second molecule.
[0217] As used herein, a "subject" may be a human or non-human
mammal or a bird. Non-human mammals include, for example, livestock
and pets, such as ovine, bovine, porcine, canine, feline and murine
mammals. In certain embodiments, the subject is human.
[0218] The term "treat," "treating" or "treatment," as used herein,
means reducing the frequency or severity with which symptoms of a
disease or condition are experienced by a subject by virtue of
administering an agent or compound to the subject.
[0219] The term "well controlled diabetes" refers to a diabetic or
pre-diabetic subject that receives treatment that allows for
keeping fasting blood sugars below 140 mg/dL. In certain
embodiments, the fasting blood sugars threshold is below 140 mg/dL,
below 130 mg/dL, below 120 mg/dL, below 110 mg/dL, or below 100
mg/dL. In certain embodiments, the fasting blood sugars range is
70-120 mg/dL. In certain embodiments, the fasting blood sugars
range is 80-100 mg/dL. In certain embodiments, the fasting blood
sugars range is 70-120 mg/dL. In certain embodiments, the fasting
blood sugars range is 70-100 mg/dL.
[0220] Throughout this disclosure, various aspects of the invention
may be presented in a range format. It should be understood that
the description in range format is merely for convenience and
brevity and should not be construed as an inflexible limitation on
the scope of the invention. Accordingly, the description of a range
should be considered to have specifically disclosed all the
possible sub-ranges as well as individual numerical values within
that range and, when appropriate, partial integers of the numerical
values within ranges. For example, description of a range such as
from 1 to 6 should be considered to have specifically disclosed
sub-ranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to
4, from 2 to 6, from 3 to 6 etc., as well as individual numbers
within that range, for example, 1, 2, 2.7, 3, 4, 5, 5.3, and 6.
This applies regardless of the breadth of the range.
[0221] Those skilled in the art will recognize, or be able to
ascertain using no more than routine experimentation, numerous
equivalents to the specific procedures, embodiments, claims, and
examples described herein. Such equivalents are considered to be
within the scope of this invention and covered by the claims
appended hereto. For example, it should be understood, that
modifications in reaction conditions, including but not limited to
reaction times, reaction size/volume, and experimental reagents,
such as solvents, catalysts, pressures, atmospheric conditions,
e.g., nitrogen atmosphere, and reducing/oxidizing agents, with
art-recognized alternatives and using no more than routine
experimentation, are within the scope of the present
application.
[0222] It is to be understood that wherever values and ranges are
provided herein, all values and ranges encompassed by these values
and ranges, are meant to be encompassed within the scope of the
present invention. Moreover, all values that fall within these
ranges, as well as the upper or lower limits of a range of values,
are also contemplated by the present application.
[0223] The following examples further illustrate aspects of the
present invention. However, they are in no way a limitation of the
teachings or disclosure of the present invention as set forth
herein.
EXPERIMENTAL EXAMPLES
[0224] The invention is now described with reference to the
following Examples. These Examples are provided for the purpose of
illustration only and the invention should in no way be construed
as being limited to these Examples, but rather should be construed
to encompass any and all variations which become evident as a
result of the teaching provided herein.
[0225] Without further description, it is believed that one of
ordinary skill in the art can, using the preceding description and
the following illustrative examples, make and utilize the compounds
of the present invention and practice the claimed methods. The
following working examples therefore, point out specific
embodiments of the present invention, and are not to be construed
as limiting in any way the remainder of the disclosure.
[0226] The materials and methods used in the experiments presented
in this Experimental Example are now described.
Example 1: Divergent Hypoglycemic Effects of Hepatic Directed
Prandial Insulin--A Six-Month Study in Type 1 Diabetes Mellitus
(T1DM)
[0227] In one aspect, HDV-I is an insulin-agnostic delivery system
that utilizes a biotin-containing lipid (such as but not limited to
biotin-phosphatidylethanolamine) in a phospholipid matrix,
targeting insulin to the liver. Mimicking portal vein delivery,
subcutaneous (SC) injection of HDV-I provides a more physiologic
treatment paradigm. Treatment with SC HDV-human regular insulin
(RHI) reduces postprandial glucose excursions compared to SC RHI.
Without wishing to be limited by any theory, HDV-I's flat
dose-response effect on hepatic glucose balance in preclinical
studies supports a fixed combination for treatment.
[0228] In the present study the use of HDV-insulin lispro (HDV-L)
vs. insulin lispro (LIS) in treating type 1 diabetes mellitus
(T1DM) was assessed. HDV-L in this study contained 1% HDV-bound LIS
and 99% unbound LIS.
[0229] ISLE-1 was a 26-week, Phase 2b, multicenter, randomized,
double-blind, non-inferiority trial. Among 176 randomized patients
(HDV-L, n=118; LIS, n=58), difference in change from baseline A1c
at Week 26 was +0.09% (95% CI -0.18% to 0.35%), confirming
non-inferiority (pre-specified margin 0.4%). Baseline A1c modified
the treatment group effect on hypoglycemia risk (interaction
p-value <0.001), with less risk of hypoglycemia (and lower
insulin dosing with similar A1c outcome) with HDV-L compared to LIS
at higher A1c, but opposite hypoglycemia effects at lower A1c
(despite similar A1c and insulin dosing). No safety signals were
identified. The present results indicate that HDV-L's hepatic
biodistribution appears to potentiate insulin effect in T1DM.
[0230] a. Design and Methods
Design and Participants:
[0231] ISLE-1 was a 26-week, Phase 2b, multicenter, randomized,
double-blind, trial in T1DM treated with multiple daily injections
(MDI) of insulin. The primary objective was A1c non-inferiority
with 26 weeks of HDV-L versus LIS.
[0232] Main inclusion criteria were: age .gtoreq.18 years; T1D for
.gtoreq.12 months; A1c .gtoreq.7.0 (.gtoreq.58 mmol/mol) to
.ltoreq.10.5% (.ltoreq.91 mmol/mol); treated with insulins glargine
or detemir for basal coverage. Main exclusion criteria were total
insulin dose .gtoreq.1.5 IU/kg/day or NPH insulin as basal.
Procedures:
[0233] Participants were randomized 2:1 ([HDV-L:LIS), stratified by
screening A1c (<8.5% [69 mmol/L] vs. .gtoreq.8.5%). Study
medications were HDV-L (0.8 ml HDV solution in 10 ml commercial
LIS) and comparator, LIS (comparably diluted with water).
[0234] Prandial dosing of HDV-lispro or control lispro was 15
minutes prior to the meal, and basal insulins were administered
either as a single daily dose or a divided twice per day dosing,
every 12 hours.
[0235] Informed of .about.10% dilution, participants continued
their current insulin parameters. Hypoglycemia was recorded on Case
Report Forms (CRFs) based on subject diaries and SMBG records,
subjectively investigator-judged as "mild," "moderate," "severe,"
or "life threatening." Blinded continuous glucose monitoring (CGM)
(Dexcom. G4) was used for 5-7 days to assess glucose at baseline,
weeks 13 and 26. A1c, lipids, and liver enzymes were measured
approximately monthly. Liver fat content MRIs were performed in a
subset.
Statistical Analysis:
[0236] The intent to treat (ITT) population included all randomized
subjects receiving at least one dose of study treatment. Safety
analyses included all randomized subjects. A sample size of 150
with assumed A1c SD of 0.8% and assumed A1c treatment difference of
0.4% had 99.9% power for non-inferiority pre-specified 0.4% margin.
Mean A1c change was analyzed using ANCOVA within intent-to-treat
(ITT) cohort at each visit. Post hoc subgroup analyses (baseline
A1c <8.5% vs .gtoreq.8.5%) were performed, this cut point
corresponding to pre-specified randomization strata. Direct
likelihood models were used for treatment arm A1c comparisons, %
time <54 mg/dL, bolus insulin, and basal insulin within the two
A1c subgroups. Poisson regression models adjusting for site as
random effect compared "severe" hypoglycemia incidence rates within
A1c groups, testing for baseline A1c by treatment group
interaction. Event number/subject was truncated at 15, accounting
for extreme outliers.
[0237] b. Discussion
[0238] Subjects were randomly assigned HDV-L (n=118) or LIS (n=58).
62% of HDV-L patients were male, with 72% of LIS male. Mean
(.+-.SD) baseline age was 46.7.+-.14.4 (HDV-L) and 44.1.+-.15.7
(LIS). Mean (.+-.SD) baseline HbA1c was 8.12.+-.0.79 (HDV-L) and
8.22.+-.0.90 (LIS).
[0239] Mean change in A1c baseline to Week 26 was -0.09% with
(HDV-L) and -0.16% (LIS), (estimated treatment difference [ETD],
HDV-L-LIS: +0.09% [95% CI -0.18 to 0.35]), confirming HDV-L
non-inferiority. Analysis of hypoglycemia outcomes showed that
baseline A1c status modified the treatment group effect on "severe"
hypoglycemia incidence (p-value for interaction <0.001), with
less hypoglycemia in HDV-L compared to LIS with poor control but
higher risk in HDV-L with better control.
[0240] Further analyses were based on subgroups (A1c .gtoreq.8.5%
vs. <8.5%). HDV-L treated subjects with baseline A1c
.gtoreq.8.5% showed a CRF-reported incidence rate of "severe"
hypoglycemia significantly lower than LIS (69 vs. 97 events/100
person-years, p=0.03), and their percentage time <54 mg/dL
during Week 26 (FIG. 1A) showed trend for reduction (median 0.7%
vs. 2.6% for HDV-L and LIS, respectively, p=0.09). Conversely, with
baseline A1c <8.5%, CRFs reported higher incidence of "severe"
hypoglycemia with HDV-L than LIS (191 vs. 21, p=0.001), and time
<54 mg/dL during Week 26 (FIG. 1B) trended higher (median 2.0%
vs. 0.6%, p=0.16). No "life threatening" events were recorded.
[0241] Exploring these divergent hypoglycemia findings, insulin
dosing was analyzed. Subjects with A1c .gtoreq.8.5% showed similar
A1c reductions for both treatments at Week 26 (p=0.35) (FIG. 1C).
However, HDV-L treated-subjects achieved A1c reductions with
.about.25% less bolus insulin than LIS subjects (mean 0.29
U*kg.sup.-1*day.sup.-1 vs. 0.38, respectively, p=0.02), with
comparable basal doses (mean 0.38 U*kg.sup.-1*day.sup.-1 vs. 0.45,
respectively, p=0.37) at study end (FIG. 1E). HDV-L and LIS
subjects with baseline A1c <8.5% both showed little change in
A1c over time (FIG. 1D) without difference in bolus/basal insulin
dosage at endpoint (p=0.86 and 0.90 for basal and bolus,
respectively) (FIG. 1F).
[0242] Lipids remained mostly stable throughout study; however, a
significant reduction in total cholesterol with HDV-L (-6.5 mg/dL)
vs. LIS (7.3 mg/dL) was observed (ETD: HDV-L-LIS: -12.0 mg/dL [95%
CI-21.1 to -2.9, p=0.01). Liver function tests at Weeks 5 and 19
showed stable ALT/AST and bilirubin levels for both treatments. Of
21 subjects studied with MM, 4 had measurable baseline liver fat;
one subject (treated with HDV-L) showed measurable liver fat
increase (3.1% baseline; 11.4% endpoint), without other evidence of
hepatic dysfunction. No treatment-related serious adverse events
were reported.
[0243] This is the first six-month study to demonstrate efficacy
and safety of a liver-targeting rapid-acting insulin formulation in
T1DM. HDV-L was non-inferior to LIS by change in A1c, with
significant total cholesterol reduction and no treatment-related
severe adverse events. In contrast to peglispro safety results
(Jacober, et al., 2016, Diabetes Obes Metab. 18 (Suppl 2):3-16),
the present study showed no between-group difference in ALT.
[0244] In certain embodiments, administration of HDV-L provides
more physiologic insulin distribution than free insulin
administration. In other embodiments, by delivering a portion of
the SC dose directly to the liver, .about.30-60% of oral
carbohydrate is sequestered as hepatic glycogen, reducing
peripheral glucose exposure and demanding reduction in peripheral
insulin exposure.
[0245] Without wishing to be limited by any theory, less well
controlled HDV-L subjects did not meaningfully alter HDV-L doses
over time (whereas LIS was increased by .about.25%) yet experienced
less CRF-reported severe hypoglycemia and less time <54 mg/dL as
compared to LIS, without difference in A1c between or within
treatments. Without wishing to be limited by any theory,
better-controlled HDV-L subjects failed to recognize a functional
increase in insulin potency, resulting in a trend for increased
time spent <54 mg/dL and significant increase in CRF-reported
hypoglycemia, despite no difference in their insulin dosing or A1c
outcomes. The strikingly divergent hypoglycemia risk findings and
differing insulin dose adjustments observed in poor-versus
better-controlled subgroups can be unified by the hypothesis that,
by altering biodistribution of SC insulin to better include the
liver, HDV increases the functional potency of insulin in both
high- and lower-A1c subgroups.
[0246] A downstream consequence of increased glycogen storage
should be improved availability of hepatic glucose to counteract
hypoglycemia; this may have occurred with HDV-L at baseline A1c
.gtoreq.8.5%, showing both relative (compared to LIS) and absolute
reductions in time below 54 mg/dL (FIG. 1A). In contrast, the lower
A1c subgroup was apparently over-insulinized owing to the increased
functional potency of HDV-L and lacked hyperglycemic "buffer" to
limit absolute hypoglycemic risk.
[0247] The present results indicate that of HDV-L is non-inferior
to LIS and its liver-targeted component potentiates insulin effect.
HDV, when added to lispro insulin, distributes meal time glucose to
the liver and as a result lowers peripheral blood glucose. In
poorly controlled T1D subjects with HbA1c >8.5%, better glycemic
control and reduced hypoglycemia was observed, even with lowered
HDV-lispro insulin doses over the course of the study. However, in
better controlled subjects, those with HbA1c <8.5%, the
reduction in peripheral glucose load led to increased hypoglycemia
incidence and severity, believed to be due to the patients not
reducing their basal (non HDV) insulin dose. In certain
embodiments, addition of HDV to an insulin makes the insulin appear
to be more potent, necessitating a re-evaluation of the
relationship of mealtime (HDV-lispro) to basal insulin dosing,
which covers times of fasting, especially overnight.
[0248] Table 1 summarizes continuous glucose monitoring results in
the Good to Great Hypoglycemia study, in terms of increased
HDV-related hypoglycemia events.
TABLE-US-00001 TABLE 1 Hypo- Treatment glycemia Difference median
HDV - (quartiles) Baseline Endpoint Lispro p-val* % Time <70
12.0% 5.8% 8.3% 5.6% +3.4% 0.01 mg/dL (4.5%, (3.1%, (6.6%, (3.3%,
(+0.7% 16.0%) 10.7%) 10.6%) 8.4%) to +6.0%) % Time <54 4.4% 2.1%
3.3% 1.7% +1.4% 0.04 mg/dL (2.5%, (0.6%, (2.0%, (0.8%, (+0.1% 8.2%)
4.1%) 5.7%) 3.3%) to +3.0%) Area above 1.7 0.9 1.2 0.7 +0.5 0.02
curve <70 (0.9, (0.3, (0.9, (0.3, (+0.1 mg/dL 2.7) 1.6) 1.9)
1.3) to +1.0) Area above 0.4 0.1 0.3 0.1 +0.1 0.07 curve <54
(0.3, (0.0, (0.2, (0.1, (-0.01 mg/dL 0.8) 0.3) 0.6) 0.3) to +0.3)
*Based on a direct likelihood model adjusting for baseline value
and a random site effect.
Example 2: Exploratory Randomized Open-Label 2-Arm Comparison of
Different Insulin Dosing Algorithms using Hepatic Directed Vesicle
(HDV)-Insulin Lispro and Insulin Degludec to Determine Optimum
Basal Insulin Dosing Regimens
[0249] The current standard of care for diabetes treatment
comprises 1:1 doses of bolus insulin and basal insulin. The present
study aims to explore the possibility of varying the ratio of
HDV-containing bolus insulin and basal insulin, so as to identify a
dosing algorithm that allows for good control of blood glucose
levels without causing hypoglycemia.
[0250] The present study is an open-label, multiple dose safety,
tolerability, and efficacy study. The study subjects are afflicted
with Type I diabetes mellitus. There is a run-in phase where all
subjects receive Insulin Lispro (HUMALOG.RTM.) for 8 weeks and then
are randomized to two groups receiving HDV-formulated Insulin
Lispro+Insulin Degludec dose.
[0251] In certain embodiments, the subject in one group receives a
dose of Insulin Degludec that is about 10% lower than the
conventional dose of Insulin Degludec used in diabetes treatments
(which would have been the same dose as the bolus insulin received
under the 1:1 paradigm).
[0252] In certain embodiments, the subject in another group
receives a dose of Insulin Degludec that is about 40% lower than
the conventional dose of Insulin Degludec used in diabetes
treatments (which would have been the same dose as the bolus
insulin received under the 1:1 paradigm).
[0253] In certain embodiments, HDV-insulin enables hepatic
metabolism of ingested carbohydrate (glucose), reducing the glucose
load to peripheral tissues, thus requiring an adjustment of basal
doses of insulin so that fasting hypoglycemia is reduced or
eliminated. The present invention provides, in one aspect, a new,
physiologically adjusted ratio of meal-time bolus HDV-insulin dose
to the 24-hour basal insulin, such as but not limited to
degludec.
Inclusion Criteria:
[0254] 1. Male or female of age 18 to 65 years, inclusive. [0255]
2. TIDM.gtoreq.12 months [0256] 3. C-peptide <0.6 ng/mL (single
retest allowed) [0257] 4. Treatment with rapid analog insulin for
the previous 6 months [0258] 5. Not using insulin pump delivery
systems during the previous 2 months [0259] 6. Use of personal
continuous glucose monitoring (CGM) technology for three months
prior to starting study and willingness to continue its use
throughout study [0260] 7. BMI .gtoreq.18.0 kg/m.sup.2 and
.ltoreq.33.0 kg/m.sup.2 10.6.5%.ltoreq.A1c.ltoreq.8.5%
[0261] The present study comprises two arms, following a 3 month
run-in period where all subjects are brought to standard of care
with insulin lispro without HDV with full characterization of their
metabolic status including HbA1c, and incidence and severity of
hypoglycemia. The first arm comprises (a) HDV-Insulin
Lispro+Insulin Degludec dose reduced by 40%. The second arm
comprises (b) HDV-Insulin Lispro+Insulin Degludec dose reduced by
10%. The primary outcome measure comprises basal, bolus, and total
insulin doses and basal/bolus ratios during the last 2 weeks of the
treatment period. The study spans 22 weeks approximately and
documents the safety and efficacy of the addition of HDV to
meal-time lispro, and the improved dosing of basal degludec insulin
to minimize the incidence and severity of hypoglycemia and
improving HbA1c levels.
[0262] During the study, the subjects are monitored for blood
glucose level, including signs of hypoglycemia. The amounts of
bolus and basal insulins provided to the subjects are then titrated
so as to ensure god glycemic control without occurrence of
significant hypoglycemia. This may involve reduction or increase in
doses of basal insulin administered to the subject, depending on
the measured biological markers.
Example 3: Hepatic Insulin Delivery to Minimize Hypoglycemic Events
in Persons with Type-1 Diabetes
[0263] Subcutaneous (SC) insulin is non-physiologic, since
pancreatic insulin goes first to the liver. The present study was
designed to determine whether delivery of Hepatic Directed Vesicles
(HDV) admixed with lispro/HUMALOG.RTM. (HDV-L) decreases
hypoglycemia in well controlled patients on multiple daily
injections (MDI) with type 1 diabetes (T1D) using unblinded Dexcom
G6 continuous glucose monitoring (CGM).
[0264] This study was a 6-month (mo) open label study of prandial
insulin (lispro, 3 mo, then HDV-L, 3 mo) with basal insulin
degludec (TRESIBA.RTM.) and unblinded continuous glucose monitoring
(CGM) in T1D with baseline A1C 6.5-8.5%. Insulin dosing,
hypoglycemia, and daily glucose control were among the monitored
parameters.
[0265] In this study the target fasting blood glucose was 80-100
mg/dL. At 3 mo subjects were randomized to -10% or -40% basal dose
to encourage titration with HDV-L. Physicians titrated basal
insulin weekly. A hypoglycemic event was defined as .gtoreq.15 min
of CGM .ltoreq.54 mg/dL.
[0266] Insulin Dosing:
[0267] At study end, degludec dosage was similar, while HDV-L dose
increased 0.03 U/kg/day (+13%, p=0.023) compared to optimal
lispro.
[0268] There was no change in basal insulin between optimal
standard of care and optimal HDV treatment, however there were
significant increases in bolus insulin dosing between optimal
standard of care and optimal HDV treatment: for the -10% group:
+0.02 U/kg/day; for the -40% group: +0.06 U/kg/day. Basal insulin
ratio was inverted in the -40% treatment group to more bolus than
basal insulin.
[0269] A1c:
[0270] In 61 enrollees, the mean baseline A1C (%) was 7.3. A1C was
6.9 after 3 mo lispro optimization, and 7.0 after 3 mo HDV-L
optimization. No significant change in A1C between optimal standard
of care and optimal HDV treatment was thus observed.
[0271] Mean Daily Glucose:
[0272] No change in mean daily glucose (<5 mg/dL) over 24 hr
day, at night or during the day.
[0273] Hypoglycemic Events:
[0274] At baseline there were 1.11 hypoglycemic events per week
(EPW) (1.04 Daytime "DT" and 1.39 Nighttime "NT" EPW), which
decreased by 11% to 0.99 EPW (0.93 DT and 1.10 NT EPW) at 3 mo. At
end of study, the switch to HDV-L resulted in a further 20%
decrease in events to 0.80 EPW (p=0.18; 0.86 DT, and 0.75 NT EPW
p=0.08).
[0275] Both -10% and -40% treatment groups demonstrated a decrease
in hypoglycemic events per week. -40% group consistently had
greater benefit -24 hour: -26% vs. -13%; Night Time: -42% vs. -21%;
Day Time: -17% vs. +1%.
[0276] Weight:
[0277] Weight (-40% group lost 0.5 kg at the end of the study)
[0278] The switch to HDV-L from lispro reduced hypoglycemia
numerically, especially nocturnally, without a significant further
change in A1C. This further hypoglycemia reduction is consistent
with the putative benefit of targeting insulin to the liver by
inducing glycogen storage postprandially, which may lead to
decrease in hypoglycemia especially at night. In certain
embodiments, by changing the bolus to basal insulin ratio (such as,
by decreasing the basal dose and increasing the bolus dose), the
patient can achieve an overall reduction in hypoglycemia events. In
other embodiments, by changing the bolus to basal insulin ratio
(such as, by decreasing the basal dose and increasing the bolus
dose), the patient can simultaneously reduce HbA1C, total
cholesterol, weight, and incidence of serious hypoglycemia events.
It is thus concluded that hepatic-directed insulin delivery in
persons with T1D helps to restore hepatic physiology.
Enumerated Embodiments:
[0279] The following exemplary embodiments are provided, the
numbering of which is not to be construed as designating levels of
importance.
[0280] Embodiment 1 provides a method of optimizing the amount of
bolus insulin and basal insulin to be administered to a subject
having diabetes mellitus and/or a metabolic derangement, wherein
the subject is administered an amount of a bolus insulin HDV
composition comprising a lipid-based nanoparticle, wherein the
bolus insulin is dispersed within the nanoparticle, wherein the
subject is further administered an amount of basal insulin, the
method comprising varying the administered amount of the bolus
insulin HDV composition and the administered amount of the basal
insulin so as to identify the optimized amount of the bolus insulin
HDV composition and the optimized amount of the basal insulin to be
administered to the subject to afford therapeutically effective
blood glucose control without significant hypoglycemia; wherein the
nanoparticle is enclosed by a bipolar lipid membrane comprising
cholesterol, dicetyl phosphate, an amphipathic lipid, and a
hepatocyte receptor binding molecule; wherein the amphipathic lipid
comprises at least one selected from the group consisting of
1,2-distearoyl-sn-glycero-3-phosphocholine,
1,2-dipalmitoyl-sn-glycerol[3-phospho-rac-(1-glycerol)],
1,2-distearoyl-sn-glycero-3-phosphoethanolamine,
1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-(succinyl),
1,2-dimyristoyl-sn-glycero-3-phosphate,
1,2-dimyristoyl-sn-glycero-3-phosphocholine,
1,2-distearoyl-sn-glycero-3-phosphate,
1,2-dipalmitoyl-sn-glycero-3-phosphate, and
1,2-dipalmitoyl-sn-glycero-3-phosphocholine; wherein the at least
one hepatocyte receptor binding molecule extends outward from the
nanoparticle; and wherein the size of the nanoparticle ranges from
about 10 nm to about 150 nm.
[0281] Embodiment 2 provides the method of Embodiment 1, wherein
the optimized amount of basal insulin to be administered to the
subject to afford therapeutically effective blood glucose control
without significant hypoglycemia is lower when the subject is
administered the bolus insulin HDV composition as compared to when
the subject is administered bolus insulin which is not part of a
HDV composition.
[0282] Embodiment 3 provides the method of any of Embodiments 1-2,
wherein the optimized amount of bolus insulin to be administered to
the subject so as to afford therapeutically effective blood glucose
control without significant hypoglycemia is lower when the subject
is administered the bolus insulin HDV composition as compared to
when the subject is administered bolus insulin which is not part of
a HDV composition.
[0283] Embodiment 4 provides the method of any of Embodiments 1-3,
wherein the insulin ratio between the optimized administered bolus
insulin HDV composition and the optimized administered basal
insulin is a function of the subject's HbA1c level.
[0284] Embodiment 5 provides the method of any of Embodiments 1-4,
wherein the insulin ratio between the optimized administered bolus
insulin HDV composition and the optimized administered basal
insulin is equal to or lower than 1:1 when the subject has >8.5%
HbA1c.
[0285] Embodiment 6 provides the method of any of Embodiments 1-4,
wherein the insulin ratio between the optimized administered bolus
insulin HDV composition and the optimized administered basal
insulin is equal to or higher than 1:1 when the subject has
<8.5% HbA1c.
[0286] Embodiment 7 provides the method of any of Embodiments 1-4,
wherein the insulin ratio between the optimized administered bolus
insulin HDV composition and the optimized administered basal
insulin ranges from about 1:0.6 to about 1:0.9 when the subject has
<8.5% HbA1c.
[0287] Embodiment 8 provides a method of optimizing the amount of
bolus insulin and basal insulin to be administered to a subject
having diabetes mellitus and/or a metabolic derangement, wherein
the subject is originally administered an amount of bolus insulin
and an amount of basal insulin such that the diabetes is well
controlled in the subject, the method comprising reducing the
amount of basal insulin administered to the subject and varying the
administered amount of a bolus insulin HDV composition so as to
identify the optimized amount of the bolus insulin HDV composition
and the optimized amount of the basal insulin to be administered to
the subject such that the diabetes is well controlled in the
subject; wherein the bolus insulin HDV composition comprises a
lipid-based nanoparticle, wherein the bolus insulin is dispersed
within the nanoparticle, wherein the nanoparticle is enclosed by a
bipolar lipid membrane comprising cholesterol, dicetyl phosphate,
an amphipathic lipid, and a hepatocyte receptor binding molecule;
wherein the amphipathic lipid comprises at least one selected from
the group consisting of 1,2-di
stearoyl-sn-glycero-3-phosphocholine,
1,2-dipalmitoyl-sn-glycerol-[3-phospho-rac-(1-glycerol)],
1,2-distearoyl-sn-glycero-3-phosphoethanolamine,
1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-(succinyl),
1,2-dimyristoyl-sn-glycero-3-phosphate,
1,2-dimyristoyl-sn-glycero-3-phosphocholine, 1,2-di
stearoyl-sn-glycero-3-phosphate,
1,2-dipalmitoyl-sn-glycero-3-phosphate, and
1,2-dipalmitoyl-sn-glycero-3-phosphocholine; wherein the at least
one hepatocyte receptor binding molecule extends outward from the
nanoparticle; and wherein the size of the nanoparticle ranges from
about 10 nm to about 150 nm.
[0288] Embodiment 9 provides the method of Embodiment 8, wherein
the subject has about 6.5-8.5% A1C.
[0289] Embodiment 10 provides the method of any of Embodiments 8-9,
wherein the subject has 80-100 mg/dL fasting blood sugar.
[0290] Embodiment 11 provides the method of any of Embodiments
8-10, wherein the subject experiences fewer hypoglycemia as
compared to the treatment without HDV.
[0291] Embodiment 12 provides the method of any of Embodiments
8-11, wherein the reduction in the amount of bolus insulin ranges
from about 1% to about 80%.
[0292] Embodiment 13 provides the method of any of Embodiments
8-12, wherein the reduction in the amount of bolus insulin ranges
from about 10% to about 40%.
[0293] Embodiment 14 provides the method of any of Embodiments
8-13, wherein the subject experiences weight loss as compared to
the treatment without HDV.
[0294] Embodiment 15 provides the method of any of Embodiments
8-14, wherein the subject does not experience significant
iatrogenic hyperinsulinemia.
[0295] Embodiment 16 provides the method of any of Embodiments
8-15, wherein the basal insulin HDV composition further comprises a
GLP-1 agonist and/or serotonin.
[0296] Embodiment 17 provides the method of any of Embodiments
8-16, wherein the GLP-1 agonist comprises liraglutide, semaglutide,
or repaglinide.
[0297] Embodiment 18 provides the method of any of Embodiments
8-17, wherein the basal insulin is formulated in a composition
comprising a lipid-based nanoparticle, wherein the basal insulin is
dispersed within the nanoparticle; wherein the nanoparticle is
enclosed by a bipolar lipid membrane comprising cholesterol,
dicetyl phosphate, an amphipathic lipid, and a hepatocyte receptor
binding molecule; wherein the amphipathic lipid comprises at least
one selected from the group consisting of
1,2-distearoyl-sn-glycero-3-phosphocholine,
1,2-dipalmitoyl-sn-glycerol-[3-phospho-rac-(1-glycerol)],
1,2-distearoyl-sn-glycero-3-phosphoethanolamine,
1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-(succinyl),
1,2-dimyristoyl-sn-glycero-3-phosphate,
1,2-dimyristoyl-sn-glycero-3-phosphocholine,
1,2-distearoyl-sn-glycero-3-phosphate,
1,2-dipalmitoyl-sn-glycero-3-phosphate, and
1,2-dipalmitoyl-sn-glycero-3-phosphocholine; wherein the at least
one hepatocyte receptor binding molecule extends outward from the
nanoparticle; and wherein the size of the nanoparticle ranges from
about 10 nm to about 150 nm.
[0298] Embodiment 19 provides the method of any of Embodiments
1-18, wherein the basal insulin is administered continuously to the
subject over a period of at least 24 hours.
[0299] Embodiment 20 provides the method of any of Embodiments
1-19, wherein the composition is administered continuously to the
subject using a pump.
[0300] Embodiment 21 provides the method of any of Embodiments
1-20, wherein the subject has a hemoglobin A1c level equal to or
lower than 8.5%.
[0301] Embodiment 22 provides the method of any of Embodiments
1-21, wherein the subject has a hemoglobin A1c level equal to or
lower than about 8.5%, and equal to or greater than 6.5%.
[0302] Embodiment 23 provides the method of any of Embodiments
1-22, wherein the membrane further comprises at least one agent
selected from the group consisting of a stabilizer and stearoyl
lysophosphatidylcholine.
[0303] Embodiment 24 provides the method of any of Embodiments
1-23, wherein the stabilizer is selected from the group consisting
of m-cresol, benzyl alcohol, methyl 4-hydroxybenzoate, thiomersal,
and butylated hydroxytoluene
(2,6-di-tert-butyl-4-methylphenol).
[0304] Embodiment 25 provides the method of any of Embodiments
23-24, wherein the stabilizer ranges from about 10% to about 25%
(w/w) in the membrane.
[0305] Embodiment 26 provides the method of Embodiment 23, wherein
the stearoyl lysophosphatidylcholine ranges from about 5% to about
30% (w/w) in the membrane.
[0306] Embodiment 27 provides the method of any of Embodiments
1-26, wherein the insulin is covalently bound to the
nanoparticle.
[0307] Embodiment 28 provides the method of any of Embodiments
1-26, wherein the insulin is not covalently bound to the
nanoparticle.
[0308] Embodiment 29 provides the method of any of Embodiments 1-28
wherein the insulin is suspended in an aqueous solution comprising
a free dissolved insulin that is not dispersed within the
nanoparticle.
[0309] Embodiment 30 provides the method of Embodiment 29, wherein
the nanoparticle-dispersed insulin and the free dissolved insulin
are independently selected from the group consisting of insulin
lispro, insulin aspart, regular insulin, insulin glargine, insulin
zinc, extended human insulin zinc suspension, isophane insulin,
human buffered regular insulin, insulin glulisine, recombinant
human regular insulin, recombinant human insulin isophane, insulin
detemir, biphasic human insulin, and insulin deglude, and any
combinations thereof.
[0310] Embodiment 31 provides the method of any of Embodiments
1-30, wherein the amphipathic lipid comprises at least one selected
from the group consisting of
1,2-distearoyl-sn-glycero-3-phosphocholine,
1,2-dipalmitoyl-sn-glycero-3-phosphocholine,
1,2-dipalmitoyl-sn-glycero-3-[phospho-rac-(1-glycerol)],
1,2-distearoyl-sn-glycero-3-phosphoethanolamine, and
1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-(succinyl).
[0311] Embodiment 32 provides the method of any of Embodiments
1-31, wherein the hepatocyte receptor binding molecule comprises
biotin.
[0312] Embodiment 33 provides the method of Embodiment 32, wherein
the biotin-containing hepatocyte receptor binding molecule
comprises at least one selected from the group consisting of
N-hydroxysuccinimide (NHS) biotin; sulfo-NHS-biotin;
N-hydroxysuccinimide long chain biotin; sulfo-N-hydroxysuccinimide
long chain biotin; D-biotin; biocytin;
sulfo-N-hydroxysuccinimide-S--S-biotin; biotin-BMCC; biotin-HPDP;
iodoacetyl-LC-biotin; biotin-hydrazide; biotin-LC-hydrazide;
biocytin hydrazide; biotin cadaverine; carboxybiotin; photobiotin;
p-aminobenzoyl biocytin trifluoroacetate; p-diazobenzoyl biocytin;
biotin DHPE (2,3-diacetoxypropyl
2-(5-((3aS,6aR)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamido)-
ethyl phosphate); biotin-X-DHPE (2,3-diacetoxypropyl
2-(6-(5-((3aS,6aR)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanami-
do)hexanamido) ethyl phosphate); 12-((biotinyl)amino)dodecanoic
acid; 12-((biotinyl)amino)dodecanoic acid succinimidyl ester;
S-biotinyl homocysteine; biocytin-X; biocytin x-hydrazide;
biotinethylenediamine; biotin-XL; biotin-X-ethylenediamine;
biotin-XX hydrazide; biotin-XX-SE; biotin-XX, SSE;
biotin-X-cadaverine; .alpha.-(t-BOC)biocytin;
N-(biotinyl)-N'-(iodoacetyl) ethylenediamine; DNP-X-biocytin-X-SE;
biotin-X-hydrazide; norbiotinamine hydrochloride;
3-(N-maleimidylpropionyl)biocytin; ARP; biotin-1-sulfoxide; biotin
methyl ester; biotin-maleimide; biotin-poly(ethyleneglycol) amine;
(+) biotin 4-amidobenzoic acid sodium salt; Biotin
2-N-acetylamino-2-deoxy-.beta.-D-glucopyranoside;
Biotin-.alpha.-D-N-acetylneuraminide; Biotin-.alpha.-L-fucoside;
Biotin lacto-N-bioside; Biotin-Lewis-A trisaccharide;
Biotin-Lewis-Y tetrasaccharide; Biotin-.alpha.-D-mannopyranoside;
and biotin 6-O-phospho-.alpha.-D-mannopyranoside.
[0313] Embodiment 34 provides the method of Embodiment 33, wherein
the biotin-containing hepatocyte receptor binding molecule
comprises at least one selected from the group consisting of biotin
DHPE and biotin-X-DHPE.
[0314] Embodiment 35 provides the method of any of Embodiments
1-34, wherein the composition further comprises cellulose acetate
phthalate, which is at least partially bound to the therapeutic
agent dispersed within the nanoparticle.
[0315] Embodiment 36 provides the method of any of Embodiments
1-35, wherein the composition further comprises at least one
charged organic molecule bound to the therapeutic agent dispersed
within the nanoparticle, wherein the charged organic molecule is at
least one selected from the group consisting of protamines,
polylysine, poly (arg-pro-thr)n in a mole ratio of 1:1:1, poly
(DL-Ala-poly-L-lys)n in a mole ratio of 6:1, histones, sugar
polymers comprising a primary amino group, polynucleotides with
primary amino groups, proteins comprising amino acid residues with
carboxyl (COO--) or sulfhydral (S--) functional groups, and acidic
polymers.
[0316] Embodiment 37 provides the method of any of Embodiments
1-36, wherein the cholesterol ranges from about 5% to about 25%
(w/w) in the membrane.
[0317] Embodiment 38 provides the method of any of Embodiments
1-37, wherein the dicetyl phosphate ranges from about 10% to about
25% (w/w) in the membrane.
[0318] Embodiment 39 provides the method of any of Embodiments
1-38, wherein the DSPC ranges from about 40% to about 75% (w/w) in
the membrane.
[0319] Embodiment 40 provides the method of any of Embodiments
1-39, wherein the hepatocyte receptor binding molecule ranges from
about 0.5% to about 10% (w/w) in the membrane.
[0320] Embodiment 41 provides the method of Embodiment 23, wherein
the amount of the stearoyl lysophosphatidylcholine in the membrane
is about 5%-30% (w/w) of the amount of DSPC in the membrane.
[0321] Embodiment 42 provides the method of Embodiment 23, wherein
the membrane comprises one of the following: (a) cholesterol,
dicetyl phosphate, DSPC, stearoyl lysophosphatidylcholine,
m-cresol, and at least one selected from the group consisting of
biotin DHPE and biotin-X-DHPE; (b) cholesterol, dicetyl phosphate,
DSPC, m-cresol, and at least one selected from the group consisting
of biotin DHPE and biotin-X-DHPE; and (c) cholesterol, dicetyl
phosphate, DSPC, stearoyl lysophosphatidylcholine, and at least one
selected from the group consisting of biotin DHPE and
biotin-X-DHPE.
[0322] Embodiment 43 provides the method of Embodiment 23, wherein
the membrane comprises cholesterol, dicetyl phosphate, DSPC,
stearoyl lysophosphatidylcholine, m-cresol, and biotin DHPE in a %
(w/w) ratio selected from the group consisting of: (a) about
9.4:18.1:56.8:14.1:0.0:1.5; (b) about 7.7:15.0:58.6:0.0:17.4:1.3;
and (c) about 8.4:16.2:47.5:7.6:19.0:1.3.
[0323] Embodiment 44 provides the method of any of Embodiments
1-43, wherein the subject has Type 1 diabetes, Type 2 diabetes,
and/or a metabolic derangement.
[0324] The disclosures of each and every patent, patent
application, and publication cited herein are hereby incorporated
herein by reference in their entirety. While this invention has
been disclosed with reference to specific embodiments, it is
apparent that other embodiments and variations of this invention
may be devised by others skilled in the art without departing from
the true spirit and scope of the invention. The appended claims are
intended to be construed to include all such embodiments and
equivalent variations.
* * * * *