U.S. patent application number 16/819740 was filed with the patent office on 2021-02-11 for dopa decarboxylase inhibitor compositions.
The applicant listed for this patent is NeuroDerm, Ltd.. Invention is credited to Oron Yacoby-Zeevi.
Application Number | 20210038505 16/819740 |
Document ID | / |
Family ID | 1000005180849 |
Filed Date | 2021-02-11 |
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United States Patent
Application |
20210038505 |
Kind Code |
A1 |
Yacoby-Zeevi; Oron |
February 11, 2021 |
DOPA DECARBOXYLASE INHIBITOR COMPOSITIONS
Abstract
Disclosed herein are formulations containing carbidopa and
optionally levodopa, arginine, and other components that have
reduced levels of impurities and toxins, particularly degradation
productions. Also disclosed herein are methods of treatment
diseases or conditions relating to a loss of dopamine or
dopaminergic neurons using such formulations, methods of making
such formulations, and kits that include such formulations.
Inventors: |
Yacoby-Zeevi; Oron; (Moshav
Bitsaron, IL) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
NeuroDerm, Ltd. |
Rehovot |
|
IL |
|
|
Family ID: |
1000005180849 |
Appl. No.: |
16/819740 |
Filed: |
March 16, 2020 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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16008228 |
Jun 14, 2018 |
10624839 |
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16819740 |
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14645848 |
Mar 12, 2015 |
10022320 |
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16008228 |
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61990967 |
May 9, 2014 |
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61952477 |
Mar 13, 2014 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 47/26 20130101;
A61K 47/20 20130101; A61K 31/133 20130101; A61K 9/0014 20130101;
A61K 47/22 20130101; A61K 47/183 20130101; A61K 31/198 20130101;
A61K 9/0019 20130101 |
International
Class: |
A61K 9/00 20060101
A61K009/00; A61K 31/133 20060101 A61K031/133; A61K 47/26 20060101
A61K047/26; A61K 47/18 20060101 A61K047/18; A61K 47/22 20060101
A61K047/22; A61K 31/198 20060101 A61K031/198; A61K 47/20 20060101
A61K047/20 |
Claims
1. A pharmaceutically acceptable formulation comprising: (i)
levodopa, about 0.1% to about 6% by weight carbidopa, about 1% to
about 25% by weight of a component selected from the group
consisting of arginine, meglumine, and a combination thereof, and
at least one o-quinone scavenger; or (ii) about 8% to about 16% by
weight levodopa, about 1% to about 4% by weight carbidopa, about
0.1% to about 40% by weight of a component selected from arginine,
meglumine, and a combination thereof, and at least one o-quinone
scavenger, wherein the pharmaceutically acceptable formulation has
less than about 1 .mu.g/ml of hydrazine, as determined by a gas
chromatography-mass spectrometry (GCMS) method.
2. The formulation of claim 1 having less than about 0.1 .mu.g/ml
of hydrazine, as determined by a GCMS method.
3. The formulation of claim 1 having about 0.1 to about 0.5
.mu.g/ml of hydrazine.
4. The formulation of claim 1, wherein said o-quinone scavenger is
selected from the group consisting of ascorbic acid or a salt
thereof, L-cysteine, N-acetylcysteine (NAC), glutathione,
diacetylcystine and/or a salt thereof, and a combination
thereof.
5. The formulation of claim 1, further comprising: about 0.1% to
about 10% by weight ascorbic acid or a salt thereof, and a
component selected from the group consisting of about 0.01% to
about 1% by weight of NAC, about 0.01% to about 1% by weight
L-cysteine, about 0.001% to about 1% by weight glutathione, and
about 0.001% to about 1% by weight diacetylcystine or a salt
thereof.
6.-9. (canceled)
10. The formulation of claim 1 comprising about 11% to about 15% by
weight levodopa.
11. (canceled)
12. A pharmaceutically acceptable formulation comprising: (a)
carbidopa; (b) ascorbic acid or a salt thereof; and (c) one of
L-cysteine, NAC, glutathione, and diacetylcystine, or a salt
thereof, wherein said formulation has less than about 1 .mu.g/ml of
hydrazine, as determined by a GCMS method.
13. The formulation of claim 12 having less than about 0.1 .mu.g/ml
of hydrazine, as determined by a GCMS method.
14. (canceled)
15. The formulation of claim 12 comprising about 0.1% to 10% by
weight ascorbic acid or salt thereof.
16.-17. (canceled)
18. The formulation of claim 12, comprising about 0.01% to about 1%
by weight L-cysteine, or a salt thereof.
19. (canceled)
20. The formulation of claim 12 comprising about 0.1% to about 10%
by weight carbidopa.
21.-26. (canceled)
27. The formulation of claim 12, further comprising levodopa.
28. The formulation of claim 12, wherein said formulation comprises
less than 4% by weight levodopa.
29.-31. (canceled)
32. The formulation of claim 12 further comprising arginine,
meglumine, or combination thereof.
33.-34. (canceled)
35. The formulation of claim 1, further comprising a
surfactant.
36.-50. (canceled)
51. The formulation of claim 1, wherein the pharmaceutically
acceptable formulation is a liquid formulation.
52.-77. (canceled)
78. A method for treating a disease or condition associated with
loss of dopamine or dopaminergic neurons in a patient, said method
comprising administering to the patient an amount of the
pharmaceutically acceptable formulation of claim 1 in an amount
effective to said disease or condition in the patient.
79. The method of claim 78 wherein said disease is Parkinson's
disease.
80. The method of claim 78, wherein said administration is
substantially continuous.
81. (canceled)
82. A method of reducing the impurities in a formulation containing
levodopa and carbidopa, said method comprising: (a) mixing together
levodopa and/or carbidopa, and a component selected from L-arginine
or meglumine, and antioxidants; (b) add the mixture of step (a)
into pre-heated water or bring the mixture at a temperature and for
a time sufficient to dissolve the powders to form a solution; (c)
cooling the solution to room temperature.
83. The method of claim 82, further comprising: (d) adding
additional water, antioxidants, and/or Tween-80, to the solution of
step (c).
84. The method of claim 82 wherein the water in step (b) comprises
antioxidants prior to said mixing.
85.-90. (canceled)
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a continuation of U.S. patent
application Ser. No. 16/008,228, filed Jun. 14, 2018, which is a
divisional of U.S. patent application Ser. No. 14/645,848, filed
Mar. 12, 2015 (now U.S. Pat. No. 10,022,320), which claims the
benefit of U.S. Provisional Patent Application No. 61/952,477,
filed Mar. 13, 2014, and U.S. Provisional Patent Application No.
61/990,967, filed May 9, 2014, the entire contents of each of which
is hereby incorporated by reference for all purposes.
BACKGROUND
[0002] Parkinson's disease is a degenerative condition
characterized by reduced concentration of the neurotransmitter
dopamine in the brain. Levodopa (L-dopa or
L-3,4-dihydroxyphenylalanine) is an immediate metabolic precursor
of dopamine that, unlike dopamine, is able to cross the blood-brain
barrier and is most commonly used for restoring the dopamine
concentration in the brain. For the past 40 years, levodopa has
remained the most effective therapy for the treatment of
Parkinson's disease.
[0003] However, levodopa has a short half-life in plasma that, even
under best common current standard of care, results in pulsatile
dopaminergic stimulation. Long-term therapy is therefore
complicated by motor fluctuations and dyskinesia that can represent
a source of significant disability for some patients. A therapeutic
strategy that could ultimately deliver levodopa/dopamine to the
brain in a more continuous and physiologic manner would provide the
benefits of standard levodopa with reduced motor complications and
is much needed by patients suffering from Parkinson's disease and
other neurological or movement disorders (Olanow C W; Mov. Dis.
2008, 23(Suppl. 3):S613-S622). Sustained-release oral levodopa
formulations have been developed, but, at best, such preparations
have been found to be no more efficacious than standard tablets.
Continuous administration of levodopa by intraduodenal
administration or infusion has also been attempted by using
ambulatory pumps or patches. Such treatments, especially
intraduodenal, are extremely invasive and inconvenient.
[0004] The metabolic transformation of levodopa to dopamine is
catalyzed by the aromatic L-amino acid decarboxylase enzyme, a
ubiquitous enzyme with particularly high concentrations in the
intestinal mucosa, liver, brain, and brain capillaries. Due to the
possibility of extracerebral metabolism of levodopa, it is
necessary to administer large doses of levodopa leading to high
extracerebral concentrations of dopamine that cause nausea in some
patients. Therefore, levodopa is usually administered concurrently
with oral administration of a dopa decarboxylase inhibitor, such as
carbidopa or benserazide, which reduces by 60-80% the levodopa dose
required for a clinical response, and thus prevents certain of its
side effects by inhibiting the conversion of levodopa to dopamine
outside the brain.
[0005] Various oral formulations together with inhibitors of
enzymes associated with the metabolic degradation of levodopa are
well known, for example, decarboxylase inhibitors such as carbidopa
and benserazide, monoamone oxidase (MAO)-A or MAO-B inhibitors such
as moclobemide, rasagiline, selegiline, and safinamide, and
catechol-O-methyl transferase (COMT) inhibitors such as tolcapone
and entacapone. Currently available oral drugs include SINEMET.RTM.
and SINEMET.RTM.CR sustained-release tablets that include carbidopa
or levodopa; MADOPAR.RTM. tablets containing levodopa and
benserazide; and STALEVO.RTM. tablets containing carbidopa,
entacapone, and levodopa.
[0006] Carbidopa is a non-competitive inhibitor of DOPA
decarboxylase. When mixed with levodopa, carbidopa inhibits the
peripheral conversion of levodopa to dopamine. This results in
increased levodopa available for transport to the CNS. Carbidopa
also inhibits the metabolism of levodopa in the GI tract, thus,
increasing levodopa bioavailability. It is used in Parkinson's
disease to reduce the peripheral effect of dopamine. The loss of
the hydrazine functional group represents the major metabolic
pathway for carbidopa.
[0007] Hydrazine (N.sub.2H.sub.4), or its salts, are used in the
production of pharmaceutical products. It has been the cause of
severe adverse effects on the central nervous system, liver, and
kidneys. In addition to these effects, experimental animals have
also shown the following symptoms: loss of body weight, anaemia,
hypoglycaemia, fatty degeneration of the liver, and convulsions.
Hydrazine has also been shown to cause DNA damage, gene mutations,
and chromosome aberrations (Environmental health criteria No. 68
hydrazine (1987)) and has induced tumor growth in mice, hamsters,
and rats after oral, intraperitoneal, and inhalation administration
(MacEwan J D, Vernot E H, Haun C C, et al. (1981)). Hydrazine and
its salts are used in the pharmaceutical industry as an
intermediate to produce drugs with different therapeutic effects
including decarboxylase inhibitiors, antihypertensives, and
antibacterials. Since hydrazine is toxic and thought to be a
possible human carcinogen, its presence is limited in some of these
drug substances in the monographs of the European Pharmacopoeia
(Ph. Eur.).
[0008] Accordingly, there is an ongoing and urgent need for liquid
formulations and compositions that can achieve continuous
dopaminergic stimulation to treat movement disorders such as
Parkinson's disease more effectively containing a safe and
tolerable amount of hydrazine.
SUMMARY
[0009] This disclosure is directed in part to carbidopa or
carbidopa ester formulations (CD), which can include levodopa
(e.g., includes levodopa (or a levodopa ester) and carbidopa). In
certain embodiments, disclosed carbidopa or carbidopa/levodopa
formulations also include two or more antioxidants, e.g., (a)
ascorbic acid or a salt thereof (e.g., sodium ascorbate) and (b)
another antioxidant, such as cysteine or a cysteine derivative (for
example, L-cysteine or N-acetylcysteine (NAC), glutathione, or
diacetylcystine), or a sulfite (e.g., sodium sulfite). In
particular, we have discovered that CD formulations that include
two antioxidants are more stable than those containing just a
single antioxidant. Certain disclosed compositions or formulations
have improved resistance to degradation (e.g., have minimal amounts
of a degradation species, e.g., hydrazine), and/or are
significantly stable.
[0010] Accordingly, in a first aspect, the invention contemplates a
pharmaceutically acceptable formulation. In one embodiment, the
formulation includes levodopa, about 0.1% to about 6% by weight
carbidopa, about 1% to about 25% by weight of a component selected
from the group consisting of arginine, meglumine, and a combination
thereof, and at least one o-quinone scavenger. In another
embodiment, the formulation includes about 8% to about 16% (e.g.,
about 11% to 15% or about 12% to about 14%) by weight levodopa,
about 1% to about 4% by weight carbidopa, about 0.1% to about 40%
by weight of a component selected from arginine, meglumine, and a
combination thereof, and at least one o-quinone scavenger. In
either of these embodiments, the pharmaceutically acceptable
formulation can have less than about 1.0, 0.75, 0.5, 0.25, 0.1,
0.05, or 0.025 .mu.g/ml of hydrazine, e.g., as determined by a gas
chromatography-mass spectrometry (GCMS) method. In particular
embodiments, the formulation has less than about 0.1 or 0.05
.mu.g/ml of hydrazine or about 0.1 to about 0.5 .mu.g/ml of
hydrazine, e.g., as determined by a GCMS method.
[0011] The formulation may include an o-quinone scavenger selected
from the group consisting of ascorbic acid or a salt thereof,
L-cysteine, NAC, glutathione, diacetylcystine and/or a salt
thereof, and a combination thereof. The formulation may further
include about 0.1% to about 10% by weight ascorbic acid or a salt
thereof and a component selected from the group consisting of about
0.01% to about 1% by weight of NAC, about 0.01% to about 1% by
weight L-cysteine, about 0.001% to about 1% by weight glutathione,
and about 0.001% to about 1% by weight diacetylcystine or a salt
thereof.
[0012] In another aspect, the invention contemplates a
pharmaceutically acceptable formulation including (a) carbidopa
(e.g., about 0.1% to about 10% carbidopa); (b) ascorbic acid or a
salt thereof; and (c) one of L-cysteine, NAC, glutathione, and
diacetylcystine, or a salt thereof. The formulation may include
less than 1.0, 0.75, 0.5, 0.25, 0.1, 0.05, or 0.025 .mu.g/ml of
hydrazine, e.g., as determined by a GCMS method. In particular
embodiments, the formulation has less than about 0.1 .mu.g/ml of
hydrazine, less than about 0.05 .mu.g/ml of hydrazine, or about 0.1
to about 0.5 .mu.g/ml of hydrazine, e.g., as determined by a GCMS
method. The formulation may include about 0.1% to 10% (e.g., 0.3%
to about 2%, about 0.5%, about 1.0% to about 1.3%, about 1.2%, or
about 1.3%) by weight ascorbic acid. The formulation may include
about 0.01% to about 1% (e.g., about 0.1% to about 0.6%, about
0.3%, about 0.4%, about 0.5%, about 0.6%, or about 0.8%) by weight
L-cysteine, or a salt thereof. The formulation may include about
0.1% to about 10% (e.g., about 0.1% to about 6%, about 0.1% to
about 4%, about 0.6% to about 1.4%, about 1.2% to about 4%, about
0.75%, about 1.4%, about 3%, or about 3.3%) by weight carbidopa.
The formulation may include about 0.1% to about 10% (e.g., about
0.4% to about 0.6%, about 0.4% to about 1%, about 0.5%, or about
1.2%) by weight ascorbic acid, or a salt thereof. The formulation
may include about 0.01% to about 1% (e.g., about 0.1% to about 1%,
about 0.3%, about 0.4%, about 0.5%, about 0.6%, about 0.7%, or
about 0.8%) by weight L-cysteine or NAC. The composition may
include less than 4% (e.g., less than 2%, 1%, 0.5%, 0.1%, 0.05%, or
0.01%) by weight levodopa or may not include levodopa. In certain
embodiments, the composition includes levodopa (e.g., about 2% to
about 16%, about 2% to about 8%, about 8% to about 16%, about 6%,
about 12% to about 15%; about 2% to about 16%, about 12%, or about
13% by weight levodopa). The composition may further comprise
arginine, meglumine, or a combination thereof, for example, about
0.1% to about 40%, about 1% to about 25%, about 10% to about 25%,
about 12% to about 40%, about 32% to about 42%, or about 15% to
about 16% by weight arginine, meglumine, or a combination thereof.
The formulation may include the components in the following
Table:
TABLE-US-00001 Components Exemplary Amount Exemplary amount
Levodopa .sup. 0-16% 5-7% Carbidopa 0.1-6% 0.6-1.5% Arginine
0.1-40% 14-16% Ascorbic acid or Na 0.1-10% 0.3-0.7% ascorbate
L-cysteine or NAC or 0.01-1% 0.3-0.5% glutathione
[0013] In particular embodiments, the formulation includes about 2%
to about 8% by weight levodopa, about 0.1% to about 3% by weight
carbidopa, about 10% to about 25% by weight arginine, about 0.1% to
about 10% (e.g., about 0.3% to about 2%) by weight ascorbic acid or
salts thereof, and about 0.001% to about 5% by weight L-cysteine or
a salt thereof. In other embodiments, the formulation includes
about 8% to about 16% by weight levodopa, about 1% to about 4% by
weight carbidopa, about 12% to about 40% by weight of a component
selected from the group consisting of arginine, meglumine, and a
combination thereof, about 0.1% to about 10% by weight ascorbic
acid or a salt thereof, and about 0.001% to about 1% by weight
L-cysteine or a salt thereof. In either of these embodiments, the
formulation has less than about 0.5 or 0.1 .mu.g/ml hydrazine
(e.g., less than 0.05 or 0.01 .mu.g/ml hydrazine), as determined by
GCMS. The formulation may include the components in the following
Tables:
TABLE-US-00002 Components Exemplary Amount Exemplary amount
Levodopa .sup. 4-8% 5-7% Carbidopa 0.5-2% 0.6-1.5% Arginine 13-18%
14-16% Ascorbic acid 0.1-2% 0.3-0.7% L-cysteine or cysteine HCl
0.1-2% 0.3-0.5%
TABLE-US-00003 Components Exemplary Amount Exemplary amount
Levodopa 10-15% 12-15% Carbidopa 1.2-4% 2-4% Arginine/meglumine or
a 25-40% 30-38% combination thereof Ascorbic acid or sodium 0.1-2%
0.3-0.7% ascorbate L-cysteine or cysteine-HCl 0.1-1% 0.2-0.5%
[0014] In other particular embodiments, the formulation includes
about 2% to about 8% by weight levodopa, about 0.1% to about 3% by
weight carbidopa, about 10% to about 25% by weight arginine, about
0.1% to about 10% by weight ascorbic acid or a salt thereof, and
about 0.001% to about 5% by weight NAC. In other embodiments, the
formulation includes about 8% to about 16% by weight levodopa,
about 1% to about 4% by weight carbidopa, about 12% to about 40% by
weight of a component selected from the group consisting of
arginine, meglumine, and a combination thereof, about 0.1% to about
10% by weight ascorbic acid or a salt thereof, and about 0.001% to
about 1% by weight NAC. In either of these embodiments, the
formulation has less than about 0.5 or 0.1 .mu.g/ml hydrazine
(e.g., less than 0.05 or 0.01 .mu.g/ml hydrazine), as determined by
GCMS. The formulation may include the components in the following
Tables:
TABLE-US-00004 Components Exemplary Amount Exemplary amount
Levodopa .sup. 4-8% 5-7% Carbidopa 0.5-2% 0.6-1.5% Arginine 13-18%
14-16% Ascorbic acid 0.1-2% 0.3-0.7% NAC 0.1-2% 0.3-0.5%
TABLE-US-00005 Components Exemplary Amount Exemplary amount
Levodopa 10-15% 12-15% Carbidopa 1.2-4% 2-4% Arginine/meglumine or
a 25-40% 30-38% combination thereof Ascorbic acid or sodium 0.1-2%
0.3-0.7% ascorbate NAC 0.1-1% 0.2-0.5%
[0015] In particular embodiments, the formulation includes about 2%
to about 8% by weight levodopa, about 0.1% to about 3% by weight
carbidopa, about 10% to about 25% by weight arginine, about 0.1% to
about 10% by weight ascorbic acid or a salt thereof, and about
0.001% to about 5% by weight glutathione. In other embodiments, the
formulation includes about 8% to about 16% by weight levodopa,
about 1% to about 4% by weight carbidopa, about 12% to about 40% by
weight of a component selected from the group consisting of
arginine, meglumine, and a combination thereof, about 0.1% to about
10% by weight ascorbic acid or a salt thereof, and about 0.001% to
about 1% by weight glutathione. In either of these embodiments, the
formulation has less than about 0.5 or 0.1 .mu.g/ml hydrazine
(e.g., less than 0.05 or 0.01 .mu.g/ml hydrazine), as determined by
GCMS. The formulation may include the components in the following
Tables:
TABLE-US-00006 Components Exemplary Amount Exemplary amount
Levodopa .sup. 4-8% 5-7% Carbidopa 0.5-2% 0.6-1.5% Arginine 13-18%
14-16% Ascorbic acid 0.1-2% 0.3-0.7% Glutathione 0.1-2%
0.3-0.5%
TABLE-US-00007 Components Exemplary Amount Exemplary amount
Levodopa 10-15% 12-15% Carbidopa 1.2-4% 2-4% Arginine/meglumine or
a 25-40% 30-38% combination thereof Ascorbic acid or sodium 0.1-2%
0.3-0.7% ascorbate Glutathione 0.1-1% 0.2-0.5%
[0016] In particular embodiments, the formulation includes about 2%
to about 8% by weight levodopa, about 0.1% to about 3% by weight
carbidopa, about 10% to about 25% by weight arginine, about 0.1% to
about 10% by weight ascorbic acid or a salt thereof, and about
0.001% to about 5% by weight diacetylcystine or a salt thereof. In
other embodiments, the formulation includes about 8% to about 16%
by weight levodopa, about 1% to about 4% by weight carbidopa, about
12% to about 40% by weight of a component selected from the group
consisting of arginine, meglumine, and a combination thereof, about
0.1% to about 10% by weight ascorbic acid, and/or a salt thereof,
and about 0.001% to about 1% by weight diacetylcystine or a salt
thereof. In either of these embodiments, the formulation has less
than about 0.5 or 0.1 .mu.g/ml hydrazine (e.g., less than 0.05 or
0.01 .mu.g/ml hydrazine), as determined by GCMS. The formulation
may include the components in the following Tables:
TABLE-US-00008 Components Exemplary Amount Exemplary amount
Levodopa .sup. 4-8% 5-7% Carbidopa 0.5-2% 0.6-1.5% Arginine 13-18%
14-16% Ascorbic acid 0.1-2% 0.3-0.7% Diacetylcystine 0.1-2%
0.3-0.5%
TABLE-US-00009 Components Exemplary Amount Exemplary amount
Levodopa 10-15% 12-15% Carbidopa 1.2-4% 2-4% Arginine/meglumine or
a 25-40% 30-38% combination thereof Ascorbic acid or sodium 0.1-2%
0.3-0.7% ascorbate Diacetylcystine 0.1-1% 0.2-0.5%
[0017] The formulation of any of the above embodiments may include
a surfactant. The surfactant may be any of one of polysorbate 20,
40, 60, and 80, or a combination thereof. In particular
embodiments, the formulation includes about 0.01% to about 5%
surfactant (e.g., polysorbate 80) or about 0.1% to 0.5% surfactant
(e.g., polysorbate 80). In more particular embodiments, the
formulation includes about 0.3% surfactant (e.g., polysorbate
80).
[0018] The formulation of any of the above embodiments may include
about 11% to about 15% by weight levodopa. For example, the
formulation may include about 12% to about 14% by weight levodopa
(e.g., about 12% or about 13.2% levodopa).
[0019] The formulation of any of the above embodiments may include
about 0.6% to about 4%, or 0.8% to about 3%, or about 1.2% to about
4%, by weight carbidopa. For example, the formulation may include
about 2.5% to about 3.5% (e.g., about 3.0% or 3.3%) by weight
carbidopa.
[0020] The formulation of any of the above embodiments may include
about 25 to about 40% (e.g., about 32% to about 40%, about 32%, or
about 36%) by weight of a component selected from the group
consisting of arginine, meglumine, and a combination thereof. For
example, the formulation may include 32% arginine, 32% meglumine,
36% arginine, or 36% meglumine.
[0021] The formulation of any of the above embodiments may, after
storage for 1, 2, 3, 4, 6, 8, 10, 12, 15, 18, 20, or 24 hours; 1,
2, 3, 5, 7, 10, 14, 21, 28, or 30 days; 1, 2, 3, 4, 6, 9, or 12
months; or 1, 1.5, 2, 2.5, or 3 years, at 25.degree. C.,
2-8.degree. C. or at -20.degree. C., have less than about 0.1
.mu.g/ml of hydrazine, as determined by GCMS.
[0022] The formulation of any of the above embodiments may have
less than about 5% (e.g., less than 4%, 3%, 2%, 1%, 0.5%, 0.3%,
0.2%, 0.1% or 0.05%) by weight
3,4-dihydroxyphenyl-2-methylpropionic acid (Degradant RRT 1.4),
relative to the amount of carbidopa, as determined by HPLC.
[0023] The formulation of any of the above embodiments may, after
storage for 1, 2, 3, 4, 6, 8, 10, 12, 15, 18, 20, or 24 hours; 1,
2, 3, 5, 7, 10, 14, 21, 28, or 30 days; 1, 2, 3, 4, 6, 9, or 12
months; or 1, 1.5, 2, 2.5, or 3 years at 25.degree. C., have less
than 1% (e.g., less than 0.75%, 0.6%, 0.5%, 0.4%, 0.3%, 0.25%,
0.2%, 0.1%, 0.05%, or 0.01%) by weight
3,4-dihydroxyphenyl-2-methylpropionic acid, as determined by HPLC.
In other embodiments, the formulation may, after storage for 1, 2,
3, 4, 6, 8, 10, 12, 15, 18, 20, or 24 hours; 1, 2, 3, 5, 7, 10, 14,
21, 28, or 30 days; 1, 2, 3, 4, 6, 9, or 12 months; or 1, 1.5, 2,
2.5, or 3 years, at 2-8.degree. C., have less than 0.5% (e.g., less
than 0.3%, 0.2%, 0.1%, or 0.01%) by weight
3,4-dihydroxyphenyl-2-methylpropionic acid, as determined by HPLC.
In other embodiments, the formulation may, after storage for 1, 2,
3, 4, 6, 8, 10, 12, 15, 18, 20, or 24 hours; 1, 2, 3, 5, 7, 10, 14,
21, 28, or 30 days; 1, 2, 3, 4, 6, 9, or 12 months; or 1, 1.5, 2,
2.5, or 3 years, at -20.degree. C., have less than 0.2% (e.g., less
than 0.15%, 0.1%, 0.05%, or 0.01%) by weight
3,4-dihydroxyphenyl-2-methylpropionic acid, as determined by
HPLC.
[0024] The formulation of any of the above embodiments may be in a
form selected from the group consisting of liquid, gel, cream,
solid, film, emulsion, suspension, solution, and aerosol (e.g., a
liquid formulation).
[0025] In another aspect, the invention features a pharmaceutically
acceptable liquid formulation including about 4% to about 8% (e.g.,
about 6%) by weight levodopa, about 0.1% to about 1.5% (e.g., about
0.6% to about 1.4%, about 0.75%, or about 1.4%) by weight
carbidopa, about 10% to about 20% (e.g., about 15% to about 16%,
about 15.2%, or about 15.6%) by weight arginine, and about 0.1% to
about 1.5% (e.g., about 0.4 to about 1%, about 0.4% to about 0.6%,
or about 0.5%) by weight ascorbic acid or a salt thereof, e.g.,
where the formulation, after 1 day at 25.degree. C., or 30 days at
25.degree. C. or after 180 days at 25.degree. C., has less than
about 1.0, 0.75, 0.5, 0.2, 0.1, or 0.05 .mu.g/ml hydrazine, as
determined by GCMS. The formulation may further include about 0.1%
to about 0.7% (e.g., about 0.4% or about 0.5%) by weight of
L-cysteine or NAC. In a particular embodiment, the formulation
includes (a) about 0.4% to about 0.6% or about 0.4 to about 1% by
weight ascorbic acid or a salt thereof and (b) about 0.1% to about
0.7% by weight of L-cysteine or NAC. In this aspect, the
formulation may further include about 0.1% to about 0.5% (e.g.,
about 0.3%) by weight Tween-80.
[0026] In another aspect, the invention features a pharmaceutically
acceptable liquid formulation including about 8% to about 16%
(e.g., about 12% to about 15%, about 12%, or about 13.2%) by weight
levodopa, about 1% to about 4% (e.g., about 3.0% or about 3.3%) by
weight carbidopa, about 20% to about 42% (e.g., about 32% to about
42%, about 32%, or about 36%) by weight of a component selected
from the group consisting of arginine, meglumine, and a combination
thereof, and about 0.1% to about 1.5% (e.g., about 1.0% to about
1.4%, about 1.2%, or about 1.3%) by weight ascorbic acid or a salt
thereof (e.g., sodium ascorbate), e.g., where the formulation,
after 1, 2, 3, 4, 6, 8, 10, 12, 15, 18, 20, or 24 hours; 1, 2, 3,
5, 7, 10, 14, 21, 28, or 30 days; 1, 2, 3, 4, 6, 9, or 12 months;
or 1, 1.5, 2, 2.5, or 3 years at 25.degree. C., has less than about
1.0, 0.75, 0.5, 0.2, 0.1, or 0.05 .mu.g/ml hydrazine as determined
by GCMS. The formulation may further include about 0.1% to about 1%
(e.g., about 0.1% to about 0.5%, about 0.3%, or about 0.5%) of
L-cysteine or salt thereof (e.g., cysteine HCl) or NAC. In a
particular embodiment, the formulation includes about 0.1% to about
0.5% of L-cysteine or NAC, and about 1.0 to about 1.4% by weight
ascorbic acid or salt thereof.
[0027] In another aspect, the invention features a method for
treating a disease (e.g., Parkinson's disease) or condition
associated with loss of dopamine or dopaminergic neurons in a
patient (e.g., a human or non-human animal, such as a mammal). The
method includes administering to the patient an amount of the
pharmaceutically acceptable formulation of any of the above aspects
in an amount effective to the disease or condition in the patient.
The method may include substantially continuous administration of
the formulation.
[0028] In another aspect, the invention features a method of
reducing the impurities in a formulation containing levodopa and
carbidopa. The method includes (a) mixing together all powders
(levodopa and/or carbidopa, L-arginine and/or meglumine,
antioxidants); (b) adding the mixture of step (a) into pre-heated
water or bringing the mixture to a temperature and for a time
sufficient to dissolve the powders to form a solution; (c) cooling
the solution to room temperature. The method may further include
(d) adding additional water, antioxidants, and/or Tween-80, to the
solution of step (c). In certain embodiments, the water in step (b)
includes antioxidants prior to the mixing. The method may be used
to produce a formulation of any one of the above aspects.
[0029] In another aspect, the invention features a kit including
one, two, or more containers having a formulation of any one of the
above aspects, where the formulation is present in an amount
sufficient to treat a patient for a disease (e.g., Parkinson's
disease) or condition resulting from decreased dopamine for at
least 1, 2, 3, 4, or 5 days; 1, 2, 3 or 4 weeks; 1 to 12 (e.g., 1
to 2, 3, 4, 6, or 9) months; or 1, 1.5, 2, 3, 4, 5, 6, 7, 8, 9, 10,
15, 20 years, or more. In certain embodiments, the formulation is
present in separate dosages.
[0030] In certain embodiments of any of the above aspects, the
formulation, method, or kit further comprises or comprises the use,
of a second agent. The second agent may be a catechol-O-methyl
transferase (COMT) inhibitor, such as tolcapone or entacapone, or a
pharmaceutically acceptable salt thereof.
BRIEF DESCRIPTION OF THE FIGURES
[0031] FIG. 1 is a graph depicting the main impurity ("Degradant")
at retention time of about 14.5.+-.0.2 min.
[0032] FIGS. 2A and 2B are graphs depicting typical MS spectrum in
negative (FIG. 2A) and positive (FIG. 2B) mode of collected main
impurity peak from a formulation sample.
[0033] FIGS. 3A and 3B are graphs depicting typical MS/MS daughter
scan (ion M/Z=179) spectrum (FIG. 3A) and parent (ion M/Z=105)
spectrum (FIG. 3B) in collected main impurity peak from a
formulation sample.
DETAILED DESCRIPTION
[0034] Provided herein are, in an embodiment, carbidopa
formulations and levodopa/carbidopa (LD/CD) formulations that can
include, for example, one, two, or more antioxidant agents or
o-quinone scavenger agents (e.g., ascorbic acid and L-cysteine, or
ascorbic acid and N-acetylcysteine (NAC)). Such formulations,
particularly those including two antioxidants (e.g., one of them
being L-cysteine or NAC) and particularly when LD is present, can
result in substantially lower levels of hydrazine, as compared to
formulations with fewer antioxidants.
[0035] Disclosed formulations can include at least such two agents
(e.g., ascorbic acid and L-cysteine, or ascorbic acid and NAC, or
sodium ascorbate and L-cysteine, or sodium ascorbate and NAC). Such
formulations can, e.g., reduce formation of undesired degradation
products and/or provide substantially stable formulations. For
example, provided herein are formulations that include carbidopa,
having less than 0.1 .mu.g/ml or less than 0.5 .mu.g/ml hydrazine,
as determined by GCMS and/or less than 5% or less than 1% by weight
3,4-dihydrooxyphenyl-2-methylpropionic acid (relative to the amount
of carbidopa), as determined by HPLC.
[0036] By "substantially continuous" administration is meant that a
dose of the formulation being administered not administered as a
bolus, e.g., a pill taken orally or a bolus injection. For example
substantially continuous administration can involve administration
of a dosage at over a period of at least 10 minutes, 30 minutes, 1
hour, 2 hours, 4, hours, 6 hours, 8 hours, 12 hours, 15 hours, 18
hours, 21 hours, or 24 hours to administer a single dose.
Substantially continuous administration can be achieved using a
transdermal patch or a pump device that continuously administers
the formulation to a patient over time.
[0037] The term "pharmaceutically acceptable carrier" or
"pharmaceutically acceptable excipient" as used herein refers to
any and all solvents, dispersion media, preservatives,
antioxidants, coatings, isotonic and absorption delaying agents,
and the like, that are compatible with pharmaceutical
administration. The use of such media and agents for
pharmaceutically active substances is well known in the art. The
compositions can also contain other active compounds providing
supplemental, additional, or enhanced therapeutic functions.
[0038] Pharmaceutically or pharmacologically "acceptable" include
formulations, molecular entities, and compositions that do not
produce an adverse, allergic or other untoward reaction when
administered to an animal, or a human, as appropriate. For human
administration, preparations should meet sterility, pyrogenicity,
general safety and purity standards as required by, e.g., the U.S.
FDA, and the EMA.
[0039] The term "pharmaceutical composition" as used herein refers
to a composition or formulation comprising at least one active
agent as disclosed herein formulated together with one or more
pharmaceutically acceptable carriers.
[0040] The term "physiologically acceptable pH" is understood to
mean a pH of, e.g., a formulation or composition that facilitates
administration of the formulation or composition to a patient
without significant adverse effects, e.g., a pH of about 4 to about
9.8 (for example, about 4.+-.0.3 to about 9.5.+-.0.3).
[0041] "Ambient temperature" as understood by a person of skill in
the art is refers to a temperature of from about 10.degree. C. to
about 30.degree. C. In particular embodiments, ambient temperature
can be 25.degree. C.
[0042] Percentages disclosed herein are by weight unless indicated
otherwise.
Compositions and Formulations
[0043] Provided herein, in an embodiment, are formulations that
include carbidopa (and/or an ester thereof) and arginine or
meglumine (or a mixture thereof). The formulation can optionally
include levodopa (and/or an ester thereof); one, two, or more
o-quinone scavenger agents; and an antioxidant. Disclosed
formulations can further include a surfactant (e.g., polysorbate
20, 40, 60, and 80, or a combination thereof).
[0044] In an embodiment, disclosed formulations can include about
0.1% to about 10% carbidopa, e.g., about 0.5% to about 8%, about
0.6% to about 5%, about 0.1% to about 1%, about 1% to about 2%,
particularly about 0.75%, about 1.4%, or about 4%. For example, a
disclosed formulation can include about 1% to about 3% by weight,
about 2.5% to about 3.5% by weight, about 0.6% to about 4% by
weight, or about 1.2% to about 4% by weight carbidopa. In certain
embodiments, disclosed compositions include about 0.01% to about 6%
by weight carbidopa, about 0.1% to about 6% by weight carbidopa, or
about 1% about to 4% by weight carbidopa, e.g., about 0.6% to 4% or
about 1.2% to 3% or 4% by weight carbidopa.
[0045] Disclosed formulations include arginine and/or meglumine (or
a salt thereof and/or a combination thereof). For example, a
disclosed formulation can include about 0.1% to about 42%, e.g.,
about 1% to about 10%, about 12% to about 18%, about 0.1% to about
40%, about 2% to about 7%, about 3.2%, about 3.4%, about 3.6%,
about 3.7%, or about 4.6% arginine and/or meglumine. In other
embodiments, disclosed formulations include about 10% to about 20%,
about 10% to about 25%, 12% to about 18%, about 12.8%, about 14.8%,
about 15.2%, about 15.5%, or about 18.5% arginine and/or meglumine
(or a combination thereof). In certain embodiments, arginine,
meglumine, a salt thereof, or a combination thereof are present at
about 25% to about 40%, about 30% to about 38%, about 32% or about
36%.
[0046] In certain embodiments, disclosed formulations can also
include a glucose amine which can, for example, replace some or all
of the arginine present in the formulations.
[0047] Disclosed formulations described herein can optionally
include levodopa. For example, in certain embodiments disclosed
formulations include about 1% to about 20% levodopa, e.g., about 2%
to about 8%, about 4% to about 7%, about 5%, or about 6%. In other
embodiments, the formulations include about 8% to about 20%, about
8% to about 16%, about 10% to about 14%, about 11% to about 14%,
about 12%, or about 13.2%. A disclosed formulation can have a molar
ratio of carbidopa to arginine (or meglumine) of about 1:1 to about
1:25 or to about 1:35.
[0048] Disclosed formulations can include one, two, or more
anti-oxidants or o-quinone scavenger agents. For example, a
disclosed formulation can include one, two, or more of an agent
each independently selected from the group consisting of ascorbic
acid or a salt thereof (e.g., sodium ascorbate, calcium ascorbate,
potassium ascorbate, ascorbyl palmitate, and ascorbyl stearate,
particularly sodium ascorbate), and cysteine or a cysteine
derivative (e.g., L-cysteine, N-acetylcysteine (NAC), glutathione,
diacetylcystine, S-methyl-N-acetylcysteine amide, acetyl
derivatives of S-methyl-N-acetylcysteine methylhydrazide,
S-methylcysteine morpholineamide, and S-methyl-N-acetylcysteine
morpholineamide, or a salt thereof). For example, a disclosed
formulations can include an ascorbic acid, or salt thereof, and a
cysteine derivative, such as NAC.
[0049] Disclosed formulations can include other antioxidants, such
as di-tert-butyl methyl phenols, tert-butyl-methoxyphenols,
polyphenols, tocopherols, and ubiquinones (e.g., caffeic acid).
[0050] Disclosed formulations can also include a tyrosinase
inhibitor. Exemplary tyrosinase inhibitors include captopril,
methimazole, quercetin, arbutin, aloesin, N-acetylglucoseamine,
retinoic acid, .alpha.-tocopheryl ferulate, MAP (Mg ascorbyl
phosphate), substrate analogues (e.g., sodium benzoate,
L-phenylalanine), Cu.sup.++ chelators, for example, Na.sub.2-EDTA,
Na.sub.2-EDTA-Ca, DMSA (succimer), DPA (D-penicillamine),
trientine-HCl, dimercaprol, clioquinol, sodium thiosulfate, TETA,
TEPA, curcumin, neocuproine, tannin, and cuprizone.
[0051] Disclosed formulations can include ascorbic acid or salt
thereof (e.g., sodium ascorbate). For example, disclosed
formulations can include 0.1% to about 10% or more ascorbic acid
(or salt thereof), or about 0.1% to about 2%, e.g., about 0.2% to
about 1.5%, about 0.2% to about 2.0%, about 0.2% to about 2.5%,
about 0.3% to about 1.2%, e.g., about 0.4%, about 0.5%, about
0.75%, about 0.85%, or about 1.0% by weight. For example, a
disclosed formulation can include about 0.8% to about 1.3% or about
1% to about 2.5% by weight ascorbic acid or salt thereof In a
particular embodiment, a disclosed formulation can include 0.5% to
about 0.85%, or, e.g., about 0.5%, about 0.75%, about 0.85%, about
1.0%, about 1.2%, or about 1.3% by weight sodium ascorbate or
ascorbic acid.
[0052] In particular embodiments, disclosed formulations can
include a bisulfite, e.g., sodium bisulfite or other sulfite salts,
e.g., sodium hydrogen sulfite or sodium metabisulfite.
[0053] In an embodiment, disclosed formulations can include for
example, NAC, L-cysteine, diacetylcystine, and/or glutathione. In
particular embodiments, the formulations includes about 0.001% to
about 5%, about 0.01% to about 5%, or about 0.1% to about 5%, about
0.001% to about 1%, or about 0.01% to about 1%, or about 0.1% to
about 1% by weight of a compound selected from the group consisting
of NAC, L-cysteine, diacetylcystine, and/or glutathione. For
example, a disclosed formulations can include about 0.01% to about
5%, e.g., about 0.05% to about 1%, 0.1% to about 0.6%, about 0.1%,
about 0.2%, about 0.3%, about 0.4%, or about 0.5% of NAC and/or
L-cysteine. In a particular embodiment, a disclosed formulation
includes about 0.4% or 0.5% NAC. In another particular embodiment,
a disclosed formulation includes about 0.3%, about 0.4%, or about
0.5% L-cysteine.
[0054] For example, a disclosed formulations can include ascorbic
acid (or a salt thereof) and a cysteine derivative, e.g.,
L-cysteine and/or NAC. In an exemplary embodiment, a disclosed
formulation includes 0.1% to about 10% ascorbic acid (or salt
thereof) and 0.001% to about 5% or about 0.001% to about 1% by
weight L-cysteine and/or NAC and/or diacetylcystine and/or
glutathione. In particular embodiments, the composition includes
ascorbic acid and L-cysteine, sodium ascorbate and NAC, ascorbic
acid and NAC, sodium ascorbate and L-cysteine, ascorbic acid and
diacetylcystine, sodium ascorbate and diacetylcystine, ascorbic
acid and glutathione, or sodium ascorbate and glutathione.
[0055] Contemplated formulations are liquid, and can include a
surfactant. For example, polysorbate 20, 40, 60, or 80 may be
present in a disclosed formulations at about e.g., about 0.01% to
about 5%, about 0.1% to about 0.5%, e.g., about 0.3%. In particular
embodiments, polysorbate 80 is present at about 0.3%.
[0056] Such formulations or solutions can have a pH that is
pharmaceutically acceptable for subcutaneous administration, e.g.,
a pH of about 8 to about 10, for example, about 9.1 to about 9.8,
e.g., 9.2 to 9.6 at 25.degree. C.
Kits, Devices, and Articles
[0057] Contemplated herein, in part, is a patch suitable for
transdermal or subcutaneous administration of an active agent in a
formulation as disclosed herein, for example, including levodopa
and carbidopa, and arginine. Such patches can have one or more
compartments that can have the same or different formulations, for
example, one compartment can have a disclosed formulation and
another a different disclosed formulation, or a different active
formulation. A dermal patch refers to any device that is capable of
delivering one or more of the active agents forming a disclosed
formulation through the skin or mucous membrane into the a
patient.
[0058] In some embodiments, disclosed liquid formulations (e.g.,
comprising carbidopa, arginine, and optionally levodopa), can be
provided in, e.g., a pre-filled cartridge or vial suitable for use
by a patient or physician. For example, provided herein is a kit
comprising a prefilled cartridge wherein a disclosed liquid
formulation is disposed within the cartridge (e.g., a pre-filled
cartridge having a single dose or a dose suitable for a single or
multiple administration to a patient of a disclosed formulation and
optionally instructions for use). For example, provided herein is a
container, vial, pre-filled syringe or the like that can include
about 1-10 ml of a disclosed formulation. For example, a
contemplated kit can include one, two, or more pre-filled vial,
container or syringe having an amount of a disclosed liquid
formulation suitable for filling a syringe pump or patch pump,
e.g., a vial, container, or syringe having about 1-6 ml, 2-5 ml,
1-2 ml, or 4-10 ml of a disclosed formulation.
[0059] The invention also contemplates kits that include
formulations of the invention that take advantage of their
increased stability. These kits can include a supply of a
formulation of the invention sufficient for at least 1, 2, 3, 4, or
5 days; 1, 2, 3 or 4 weeks; 1, 2, 3, 4, 6, or 9 months; or 1 or 1.5
years of administration to a patient, which can be packaged, for
example, into suitable dosage (e.g., unit dosage) formulations.
These kits can optionally include instructions for their use. A kit
for daily use for example, can include one, two or more containers
or vials of a disclosed formulation, an infusion set, and a
disposable patient delivery units (e.g. syringe).
Preparation of Compositions
[0060] Disclosed formulations or compositions can be prepared by
mixing arginine and/or meglumine in amounts as disclosed above with
levodopa and/or carbidopa, and optionally anti-oxidant(s), e.g., to
form a powder mixture. Water can be added to the mixture to form a
suspension. The water can be pre-heated or the suspension can be
heated at a temperature and for a time sufficient to dissolve the
mixture, e.g., to about 40.degree. C. to about 100.degree. C., or
to about 40.degree. C. to 80.degree. C., or to about 60.degree. C.
to 90.degree. C., e.g., 65.+-.5.degree. C. or 72.+-.5.degree. C. or
73.+-.3.degree. C., e.g., by adding pre-heated water and/or by
placing the mixture in a hot (e.g., 65.+-.5.degree. C. or
72.+-.5.degree. C. or 73.+-.3.degree. C.) water bath (e.g., for up
to about 10 minutes, for about 3 minutes, for about 5 minutes, or
for about 10, 20, 30, 40, 50, 60 minutes, or more) to form a
solution, with optional stirring. This is followed by cooling the
solution to form the composition. N.sub.2 can be provided the head
space of the container. For example, the mixture can then be
removed from the hot water bath and cooled to room temperature, and
adding, e.g., immediately thereafter, an optional anti-oxidant(s)
under N.sub.2 atmosphere and subsequent stirring. A preparation
such as that above, e.g., where levodopa and/or carbidopa, and
arginine are mixed together as powders first, and a suspension
formed with water and then heated can result in a more stable
solution, as compared to a preparation that includes a stepwise
preparation of individual water suspensions of ingredients and
later combination. Specific methods of preparation are described in
Example 1 below.
[0061] Disclosed formulations can be sterilized, e.g., using 0.2
.mu.m filters such as filters with nylon or PVDF membranes. In
another embodiment, the preparation of disclosed formulations has
fewer undesirable by-products when pre-heated water is added as
disclosed above, as compared to a formulation prepared without the
addition of pre-heated water. In another embodiment, the levodopa
and/or carbidopa cannot dissolve unless the preparation procedure
disclosed is used. Such disclosed preparations as above can provide
a more stable formulation as compared to a formulation prepared
without adding hot water or heating. Disclosed preparations can
provide a more stable formulation as compared to a formulation
prepared without adding antioxidants prior to heating.
Methods of Treatment
[0062] In another aspect, the present invention contemplates a
method for treatment of a disease or disorder, such as a
neurological disorder (e.g., a disorder associated with reduced
dopamine or loss of dopaminergic neurons) or a movement disorder,
comprising administering a formulations described herein. The
method can include substantially continuously administering the
formulation.
[0063] Also provided herein are methods of treating a neurological
disorder (e.g., a disorder associated with reduced dopamine or
death of dopaminergic neurons) or a movement disorder that include
substantially continuous administration of a formulation described
herein.
[0064] Without limiting, the pharmaceuticals compositions as
described above can be used for treating a neurological disorder
(e.g., a disorder associated with reduced dopamine or death of
dopaminergic neurons) or a movement disorder that include acute and
immediate administration such as inhalation or injection.
[0065] In some embodiments, compositions comprising levodopa (e.g.
a disclosed liquid composition) may be administered at a rate of
about 0.16 ml/hour/site to about 0.24 ml/hour/site, or, e.g., about
0.01 ml/hour to about 0.4 ml/hour/site. Such rates may be constant
throughout the day and night or varied according to patient's need,
for example, may reflect a patient resting or sleeping schedule and
waking or higher activity level schedule. For example, liquid
compositions such as those disclosed herein (e.g., including
levodopa) may be administered at a rate of about 0.32 ml/hour/site
in the morning (e.g., for about 2-4 hours before waking), about
0.24 ml/hour/site during the daytime or activity time, (e.g., for
about 10 to about 12 hours), and/or about 0.08 ml/hour/site at rest
or at night. In another embodiment, liquid composition such as
those disclosed herein may be administered, e.g., intraduodenally,
at a rate of about 1.0 ml/hour during the daytime or activity time
(e.g., for about 2-3 hours before waking and for about 10 to about
12 hours thereafter), and 0 to about 0.5 ml/hour at rest or at
night. In another embodiment, liquid compositions such as disclosed
herein (e.g., comprising levodopa and arginine), may be
administered at a rate of about 1.25 ml/hour (e.g., about
1.25.+-.0.5 ml/hour during the daytime or activity time (e.g., for
about 2-3 hours before or after waking and for about 10 to about 14
hours thereafter) and 0 to about 0.5 ml/hour (e.g. about
0.5.+-.0.25 ml/hour) at rest or night. In further embodiments, such
compositions may be administered at a rate of about 0.1 to about
1000 .mu.l/hour/site; or at a volume of about 2 to about 10 ml/24
hour/site, preferably about 4 to about 6 ml/24 hour/site; or at a
dose of about 80 to about 800 mg levodopa/day and about 20 to about
200 mg carbidopa/day; or at a rate of about 240 to about 360 mg
levodopa and about 60 to about 90 mg carbidopa/day/site.
[0066] Contemplated administration, following the disclosed
methods, typically can be carried out over a defined time period
(usually weeks, months, or years). Administration can be effected
by any appropriate route including, but not limited to,
subcutaneous, oral routes, intravenous routes, intramuscular
routes, intradermal routes, subcutaneously, intratracheally,
intrathecally, intraduodenally, transdermally, inhalation, and/or
direct absorption through mucous membrane tissues.
[0067] The disease or disorder characterized by reduced levels of
dopamine in a patient contemplated herein are neurological or
movement disorders including restless leg syndrome, Parkinson's
disease, secondary parkinsonism, Huntington's disease, Shy-Drager
syndrome, and conditions resulting from brain injury including
carbon monoxide or manganese intoxication. Methods for treating
such disorders in a patient in need thereof are provided, for
example, by administering (e.g., subcutaneously) a disclosed
formulation. In one embodiment, the disease to be treated is
Parkinson's disease.
[0068] In an embodiment, substantially continuously administering
using, e.g., a liquid formulation can be via a pump for
subcutaneous infusion (insulin pump) at an average rate of about
10-1000 .mu.l/hour (e.g., 10-250 .mu.l/hour), about 300.+-.100
.mu.l/hour, or about 200.+-.40 .mu.l/hour continuously for 24
hours; about 440.+-.200 .mu.l/hour or about 200.+-.50 .mu.l/hour
continuously for 16 hours (during waking hours) and at night (e.g.,
for 8 hours, about 0 to 80 .mu.l/hour or 0 to 200 .mu.l/hour or via
a pump or a transdermal patch. Substantially continuously
administering the formulation in to a patient can be doubled or
tripled by using more than one pump or site of infusion. In an
embodiment, substantially continuously administering using, e.g., a
liquid formulation can be at an average rate of about 0.2-2
.mu.l/hour, or about 1.+-.0.5 .mu.l/hour continuously for 24 hours;
about 1.0.+-.0.5 .mu.l/hour continuously for 16 hours (during
waking hours) and at night (e.g., for 8 hours, about 0 to 0.5
.mu.l/hour via a pump or transdermal patch, or combination of
delivery devices that are suitable for, e.g., subcutaneous,
intravenous, intrathecal, and/or via the duodenum).
[0069] The invention now being generally described, it will be more
readily understood by reference to the following examples which are
included merely for purposes of illustration of certain aspects and
embodiments of the present invention, and are not intended to limit
the invention in any way.
EXAMPLES
Example 1 Formulation Preparation Procedure
[0070] Levodopa (LD) and carbidopa (CD) formulations can be
prepared as follows.
[0071] Method #1 (L-Arg solution): L-Arg and Na-Bis (Na-bisulfite)
were dissolved in water. The solution was added to the LD and CD
powders. The mixture was heated with stirring for 13 min at
75.degree. C. until fully dissolved. The LD/CD solution was kept at
room temperature (RT) for 10 min to cool down.
[0072] Method #2 (all powders together): All powders (LD, CD, and
L-Arg) were weighed, and water with Na-Bis was added. The
suspension was heated with stirring for 13 min at 75.degree. C.
until fully dissolved. The LD/CD solution was kept at RT for 10 min
to cool down.
[0073] Method #3 (same as #2 without Na-Bis pre-heating): All
powders (LD, CD, and L-Arg) were weighed together and water was
added. The suspension was heated with stirring for 13 min at
75.degree. C. until fully dissolved. The LD/CD solution was kept at
RT for 10 min to cool down.
[0074] Method #4 (preparation in steps): LD and the respective
amount of L-Arg were weighed; water and Na-Bis solution were added.
The suspension was heated for 7 min at 75.degree. C. until fully
dissolved, followed by 7 min at RT. CD and the respective amount of
L-Arg were weighed and added to the LD/Arg solution at 60.degree.
C. until fully dissolved. Finally, extra L-Arg was added.
[0075] Method #5 (same as #4 without Na-Bis pre-heating): LD and
the respective amount of L-Arg were weighed; water was added. The
suspension was heated for 7 min at 75.degree. C. until fully
dissolved followed by 7 min at RT. CD and the respective amount of
L-Arg were weighed and added to the LD/Arg solution at 60.degree.
C. until fully dissolved. Finally, extra L-Arg was added.
[0076] After cooling down, all formulations from all methods were
divided in to 3 vials, and water, Na-Bis solution, or Na-Bis-Arg
solution was added to each vial.
Example 2: Identification of the Main Degradant in Formulations
Containing Carbidopa
[0077] Liquid formulations with levodopa, carbidopa, and arginine
were prepared using the procedure outlined in Example 1, and HPLC
analysis was performed according to APH stability-indicating
analytical method for carbidopa levodopa formulations with Agilent
1100 system.
[0078] The HPLC system used herein includes the following
components manufactured by Agilent: pump system (model G1311A),
diode array detector (model G1315B), autosampler (model G1329A),
degasser (model G1379A), thermostat (model G1330B), thermostatted
column compartment (model G1316A). The column employed was a new
Synergi 4.mu. Fusion-RP 80A, 250.times.4.6 mm
(phenomenex.RTM.).
[0079] HPLC Working Conditions:
[0080] Wavelength: 280 nm
[0081] Flow rate: 1.0 ml/min
[0082] Inj. volume: 10 .mu.l
[0083] Column temperature: 30.degree. C.
[0084] Thermostat temperature: 4.degree. C.
[0085] Stop time: 27 min
[0086] Pressure: 105 bar
[0087] Mobile Phase Preparation:
[0088] Solvent A: Acetonitrile
[0089] Solvent B: 20 mM potassium dihydrogen phosphate, pH=2.4
[0090] The mobile phase B was prepared by weighing 2.72 g/L of
potassium dihydrogen phosphate. pH was adjusted to 2.4 by addition
of H.sub.3PO.sub.4.
[0091] Gradient:
TABLE-US-00010 Time Solvent A Solvent B Flow 0 5 95 1.0 5 5 95 1.0
15 60 40 1.0 20 60 40 1.0 20.01 5 95 1.2 27 5 95 1.2
[0092] Diluent:
[0093] 0.1M HCl/MeOH 9:1
[0094] (8.3 ml HCl 37% to 1L)-->0.1 M HCl
[0095] STD LDOPA=100.00 mg/100 ml
[0096] STD CDOPA=25.00 mg/100 ml
[0097] Calibration Curve:
TABLE-US-00011 STD stock sol. Volumetric Final concentr. 1000/250
ppm bottle, ml LD/CD ppm NA 10 1000/250.sup. 5000 from stock 10
500/125 5000 from 500/125 10 250/62.5 1000 from stock 10
100/25.sup. 5000 from 100/25 10 50/12.5 1000 from 100/25 10
10/2.5
[0098] One ml of the sample (levodopa/carbidopa formulation) was
transferred to a 25 ml amber volumetric glass flask and filled to
volume with diluent (0.1 M HCl/MeOH 9/1). The sample was degraded
with hydrogen peroxide.
[0099] The impurity was observed at retention time of about
14.5.+-.0.2 min (FIG. 1). To ensure that the peak observed is
actually the compound of interest, the main degradant peak was
collected from analytical HPLC, evaporated under nitrogen stream
and reconstituted with diluent. Obtained samples were tested by
HPLC/MS.
[0100] Initially, an MS scan analysis was applied (FIGS. 2A and
2B). The unknown compound shows clear and intensive signal in
negative mode and much nosier signal in positive mode. Therefore it
was expected to be more a proton donor than acceptor. The base peak
in negative mode was M/Z=195 Da that was suspected as (M-H). The
mass difference between this ion and peak M/Z=217 is 22 and that
should be the sodium adduct. This is the evidence of the presence
of carboxyl or/and phenol groups. Due to this fact, the molecular
weight was proposed to be 196 Da.
[0101] The peak M/Z=197 (M+H+)+ was not found in positive mode, but
the peak M/Z=197 (M-H2O)+ is observed. This is typical for oxygen
containing molecule.
[0102] The daughter and parent MS/MS was performed as well to
define molecular structure. Peaks observed in positive mode with
M/Z=179,161,151,133,123,105 were found to be relative. They were
defined as in-source fragment ions arising from the molecular ion
with M/Z=197. The typical MS and MS/MS spectra are shown on figures
(FIGS. 3A and 3B).
[0103] The chemical formula of the degradant compound is
C.sub.10H.sub.12O.sub.4; with a molecular structure given by 3-(3,4
dihydroxyphenyl)-2-methylpropanoic acid.
Example 3: Effect of Ascorbic Acid With or Without EDTA on LD/CD
Formulation Stability
[0104] Liquid formulations were prepared by weighing all powders
(LD, CD, EDTA, ascorbic acid, and L-Arg) and adding water
pre-heated to 73.+-.3.degree. C. The suspension was put in a water
bath at 73.+-.3.degree. C. and stirred for 10 min until fully
dissolved. LD/CD solution was kept at RT for 10 min to cool down.
Solutions were divided into glass vials and kept at +25.degree. C.
and at -20.degree. C. for the indicated period of time. Prior to
analyses, frozen vials were placed at RT until fully thawed.
Formulations were then mixed and subjected to stability analyses.
The effect of ascorbic acid with or without EDTA on stability of
LD/CD formulations was measured by HPLC. The levels of the
degradant presented in Tables A and B indicate the level of
stability of LD/CD formulations.
TABLE-US-00012 TABLE A Ca-EDTA- Ascorbic L-Arginine Na.sub.2 acid t
= 0 LD/CD (%) (%) (%) Degradant Total (%) 6/1.4 14.80 0.2 1.0 0.13
0.76 6/1.4 14.80 0 1.0 0.08 0.44
TABLE-US-00013 TABLE B L-Arginine Ca-EDTA-Na.sub.2 Ascorbic acid t
= 0 t = 1 w (25.degree.) LD/CD (%) (%) (%) Degradant Total (%)
Degradant Total (%) 6.0/1.4 14.80 0.2 1.0 0.054 0.64 0.16 0.79
6.0/1.4 14.80 0 1.0 0.046 0.49 0.15 0.49
Tables A and B indicate that EDTA did not have a significant effect
on the stability of LD/CD formulations.
Example 4: Effect of L-Cysteine on the Stability of Solutions
Containing CD
[0105] Liquid formulations were prepared by weighing all powders
(LD, CD, L-cysteine, ascorbic acid, and L-Arg) and adding water
pre-heated to 73.+-.3.degree. C. The suspension was put in a water
bath at 73.+-.3.degree. C. and stirred for 10 min until fully
dissolved. LD/CD solution was kept at RT for 10 min to cool down.
For CD formulations, CD, L-cysteine, and ascorbic acid were
weighed, and pre-heated water (60.degree. C.) was added. Solutions
were divided into glass vials and kept at +25.degree. C. and at
-20.degree. C. for the indicated period of time. Prior to analyses,
frozen vials were placed at RT until fully thawed. Formulations
were then mixed and subjected to stability analyses. The effect of
L-cysteine on the stability of carbidopa formulations when stored
at 25.degree. C., either exposed to air or maintained under
anaerobic conditions (N.sub.2), was analyzed using HPLC. The levels
of the degradant presented in Table C point toward the level of
stability of carbidopa formulations.
TABLE-US-00014 TABLE C Ascorbic L- Degradant L- acid Cysteine t = 5
t = 5 LD/CD arginine (%) (%) t = 0 days weeks 6/1.4 14.8 0.5 0.1
N.sub.2 0.08 0.22 0.28 O.sub.2 0.59 0.77 0.5 0.2 N.sub.2 0.08 0.20
0.25 O.sub.2 0.36 0.32 0.5 0.4 N.sub.2 0.00 0.14 0.14 O.sub.2 0.14
0.17 0/4.sup. 4.6 0.5 0.1 N.sub.2 0.05 0.16 O.sub.2 1.42 0.5 0.2
N.sub.2 0.05 0.16 O.sub.2 0.56 0.5 0.4 N.sub.2 0.04 0.15 O.sub.2
0.35
[0106] As Table C indicates, ascorbic acid with 0.1% L-cysteine was
sufficient to inhibit degradant formation in formulations
containing carbidopa (with or without levodopa) when kept under
anaerobic conditions at 25.degree. C. for at least 5 weeks. We can
further deduce from Table C, that L-cysteine reduced degradant
formation under aerobic conditions at 25.degree. C. in a
dose-dependent manner.
[0107] In formulations containing carbidopa and 0.4% L-cysteine,
degradant formation was inhibited during the preparation of the
formulation. These formulations were stable for at least 5 weeks at
25.degree. C. under both aerobic and anaerobic conditions.
Formulations containing carbidopa are more stable when they also
contain LD upon exposure to air.
Example 5: Effect of L-Cysteine on 6/1.4% Levodopa/Carbidopa
Formulation Stability
[0108] Liquid formulations were prepared as described in Example 3.
The effect of L-Cysteine on the stability of 6/1.4%
levodopa/carbidopa solutions at 25.degree. C. was analyzed using
HPLC. The levels of the degradant presented in tables D, E, F, and
G point toward the level of stability of referenced
formulations.
TABLE-US-00015 TABLE D LD/CD L-Arginine Ascorbic Acid L- Cysteine
Degradant (%) (%) (%) (%) t = 0 6/1.4 14.8 0.5 0.2 0.02 0.75 0.0
0.16
TABLE-US-00016 TABLE E L- Ascorbic L- LD/CD arginine acid Cysteine
Degradant (%) (%) (%) (%) t = 0 t = 5 d t = 5 wk 6/1.4 14.8 0.75 0
0.49 0.93 1.08 0.1 0.10 0.35 0.32 0.50 0.1 0.16 0.28 0.45 0.4 0.00
0.15 0.13
TABLE-US-00017 TABLE F L- Ascorbic L- LD/CD arginine acid Cysteine
Degradant (%) (%) (%) (%) t = 0 t = 1 d t = 4 d 6/1.4 14.8 0.5 0.4
0.00 0.00 0.27 0.1 0.25 0.69 1.26 0.75 0.1 0.23 0.56 0.85 15.1 1.0
0.2 0.00 0.26 0.51
TABLE-US-00018 TABLE G LD/CD L-arginine Ascorbic acid L-Cysteine
Degradant (%) (%) (%) (%) t = 0 t = 10 d t = 3 weeks t = 3 months
6/1.4 14.8 0.5 0.1 0.15 0.43 0.46 0.87 0.75 0.0 1.01 1.40 1.37
2.00
[0109] Results show that levodopa/carbidopa formulations were more
stable with both ascorbic acid and L-cysteine, as compared to
ascorbic acid alone, suggesting that L-cysteine and ascorbic acid
have a synergistic effect in preventing degradant formation. Other
results showed that L-cysteine alone had no effect at all (data not
shown). Furthermore, L-cysteine inhibited degradant formation
during formulation preparation and maintained the stability of the
formulation for at least up to 5 weeks at 25.degree. C., in a dose
dependent manner. Increasing the amount of ascorbic acid reduces
degradant formation, but this was significantly less efficient than
the combination of ascorbic acid with L-cysteine.
Example 6: Effect of Tween-80 and Na-Ascorbate on
Levodopa/Carbidopa Formulation Stability
[0110] Liquid formulations were prepared by weighing all powders
(LD, CD, L-Cysteine, ascorbic acid, Na-ascorbate and L-Arg) and
adding water pre-heated to 73.+-.3.degree. C. The suspension was
put in a water bath at 73.+-.3.degree. C. and stirred for 10 min
until fully dissolved. LD/CD solution was kept at RT for 10 min to
cool down. Then, Tween-80 was added. Solutions were divided in to
glass vials and kept at +25.degree. C. and at -20.degree. C. for
the indicated period of time. Prior to analyses, frozen vials were
placed at RT until fully thawed. Formulations were then mixed and
subjected to stability analyses. The effect of Tween-80 and
Na-ascorbate on the stability of carbidopa/levodopa formulations
was analyzed using HPLC. The levels of the degradant presented in
table H point toward the level of stability of carbidopa/levodopa
formulations.
TABLE-US-00019 TABLE H LD:CD L-Arginine Asc. acid:L-Cys Tw-80
Degradant (%) (%) (%) (%) t = 0 t = 1 month 6.0:1.5 14.8 0.5:0.4 0
0.07 0.08 6.0:1.5 14.8 0.5:0.4 0.3 0.10 N/A 6.0:1.5 14.8 0.5:0.4
0.75 0.15 6.0:1.5 14.8 0.5:0.4 2.0 0.00 6.0:1.5 14.8 0.75:0.1 0
0.26 6.0:1.5 14.8 0.75:0.1 0.75 0.31 6.0:1.5 14.8 0.75:0.2 0 0.14
6.0:1.5 14.8 0.75:0.2 0.3 0.27 6.0:1.5 14.8 0.75:0.2 0.75 0.24
6.0:1.5 14.8 0.75:0.2 2.0 0.35 7.5:1.5 18.5 0.75:0.2 0 0.19 0.26
7.5:1.5 18.5 0.75:0.2 0.75 0.35 0.45 7.5:1.5 18.5 0.85*:0.2 0 0.23
0.20 7.5:1.5 18.5 0.85*:0.2 0.75 0.25 0.27 *Sodium ascorbate
[0111] The results demonstrate that Tween-80 did not have an effect
on degradant formation. It is also shown that the effect of
L-cysteine on the stability of the formulations was dose dependent.
Table H further shows that the effect of Na-ascorbate and ascorbic
acid on the stability of the formulations and degradant formation
was similar.
Example 7: Effect of Ascorbic Acid With or Without L-Cysteine or
NAC on Long Term Stability of Levodopa/Carbidopa Formulations
[0112] Liquid formulations were prepared by weighing all powders
(LD, CD, arginine, L-Cysteine or NAC, and ascorbic acid or
Na-ascorbate) and by adding water pre-heated to 73.+-.3.degree. C.
The suspension was put in a water bath at 73.+-.3.degree. C. and
stirred until fully dissolved. LD/CD solution was kept at RT to
cool down. Then Tween-80 was added. Solutions were divided into
glass vials and kept at +25.degree. C. and at -20.degree. C. for
the indicated period of time. Prior to analyses, frozen vials were
placed at RT until fully thawed. Formulations were then mixed and
subjected to stability analyses. The effect of ascorbic acid with
or without L-cysteine or NAC on the stability of carbidopa/levodopa
formulations was analyzed using HPLC. The levels of the degradant
presented in Table I indicate the level of stability of
carbidopa/levodopa formulations.
TABLE-US-00020 TABLE I Degradant Ascorbic Tween- 2 .times. F - T*
LD/CD acid (NAC) 80 Arginine -20.degree. C. 25.degree. C. (ambient)
(%) (%) (%) (%) (%) T.sub.0 1 m 2 m 3 m 6 m 9 m 12 m 16.5 m 1 m
>3 m/>7 d 5/1.15 0.75 0 0 12.8 0.55 0.6 0.75 0.9 1.0 0.8 --
1.0 1.2 -- 6/1.4 0.75 0 0 14.8 0.4 0.45 0.45 0.6 0.5 0.8 -- 0.6 0.6
-- 6/1.4 0.5 0.4 0 14.8 0 0 -- -- -- 0.25 0.15 -- 0 -- 6/1.4 0.5
0.4 0.3 15.5 0 -- 0.1 0.2 0.2 0.2 -- -- -- -- 6/1.4 0.5 (0.5) 0.3
15.5 0 0 -- -- -- -- -- -- 0.2 0 6/0.75 0.5 (0.5) 0.3 15.2 0 0.1 --
-- -- -- -- -- 0 0.2 *2 .times. F/T--at least 2 freeze-thaw cycles
after >3 m at -20.degree. C., and >7 d at ambient temp.
[0113] The results presented in Table I suggest that formulations
containing both ascorbic acid and L-cysteine or NAC are more stable
than formulations having only ascorbic acid, at a) T.sub.0, i.e.,
immediately following formulation preparation, b) for at least 9
months at -20.degree. C., and c) at for least 1 month at ambient
temperature.
Example 8: Effect of Antioxidants on the Stability of Carbidopa
Formulations
[0114] Liquid formulations with carbidopa and arginine were
prepared as described above. The effect of antioxidants on the
stability of carbidopa formulations was analyzed using HPLC. The
levels of the degradant presented in Tables J and K point toward
the level of stability of carbidopa formulations.
TABLE-US-00021 TABLE J 1 2 3 4 7 weeks at 0.1% Na 0.075% Na 0.4%
Asc. + 0.5% Asc. + (-20.degree. C.) Bisulfite Bisulfite 0.2% L-Cys
0.1% L-Cys Methyl-Dopa 0.15 0.15 0.15 0.15 Unknown 2 0.18 0.17 0 0
Degradant 0.54 0.55 0 0.16 Sum of all 1.14 1.15 0.44 0.65
Impurities
TABLE-US-00022 TABLE K Peak 14.3 min (degradant) t = 3 weeks t = 3
weeks t = 0 2-8.degree. C. 25.degree. C. 2 4% CD - 3.7% L-Arg 0.30
1.12 .+-. 0.19 1.08 .+-. 0.07 0.075% sodium bisulfite 3 4% CD -
4.62% L-Arg 0.06 0.15 .+-. 0.01 0.29 .+-. 0.07 0.4% ascorbic + 0.2%
L-cysteine 4 4% CD - 4.62% L-Arg 0.11 0.44 .+-. 0.21 0.63 .+-. 0.39
0.5% ascorbic + 0.1% L-cysteine
[0115] The results in Tables J and K suggest that formulations
containing ascorbic acid +L-cysteine were significantly more stable
than the formulation containing Na-bisulfite (formulations 3 &
4 vs. formulations 1 & 2). The same amount of impurities were
measured with 0.075 and 0.1% Na-bisulfite, suggesting that the
maximum possible protection with Na-bisulfite was attained.
[0116] In sum, the combination of ascorbic acid/L-cysteine is able
to prevent degradant and other impurities formation, such as
hydrazine (see other examples), while Na-bisulfite does not protect
formulations containing carbidopa to the same extent.
Example 9: Effect of Various Antioxidants and Different
Concentrations of Arginine on the Stability of Formulations
Containing 4% CD
[0117] Liquid formulations with carbidopa and arginine (Table L)
were prepared as described above. The effect various antioxidants
and different concentrations of arginine on the stability of
formulations containing 4% CD, and stored under aerobic (air) or
anaerobic (N.sub.2) conditions, at ambient (25.degree. C.) or cold
(2-8.degree. C.) temperature was evaluated using HPLC analysis. The
levels of the degradant and total impurities as presented in Tables
M and N, respectively, point toward the level of stability of
carbidopa formulations.
TABLE-US-00023 TABLE L 1 2 3 Carbidopa 4.0 4.0 4.0 L-Arginine 4.6
4.6 3.7 Ascorbic acid 0.4 0 0 L-Cysteine 0.2 0 0 Na bisulfite (5%)
0 1.5 1.5 Total solutes 9.2 10.1 9.2 Water 90.8 89.9 90.8 L-Arg
(mM) 265 265 212 CD (mM) 177 177 177 Ratio L-Arg/CD 1.5 1.5 1.2 pH
measured 8.67 8.87 8.62
TABLE-US-00024 TABLE M degradant: Peak area (%) T = 1 week T = 1
week 2-8.degree. C. 25.degree. C. t = 0 Air N2 Air 1 0.4% ascorbic
+ 0.2% cysteine 0.1 1.7 0.2 5.8 (CD:Arg 1.0:1.5) 2 0.075% Bisulfite
0.2 1.9 1.2 6.2 (CD:Arg 1.0:1.5) 3 0.075% Bisulfite 0.25 2.4 1.2
8.3 (CD:Arg 1.0:1.2)
TABLE-US-00025 TABLE N Total degradation products: Peak area (%) T
= 1 week T = 1 week 2-8.degree. C. 25.degree. C. t = 0 Air N2 Air 1
0.4% ascorbic + 0.2% cysteine 0.1 2.5 1.1 7.2 (CD:Arg 1.0:1.5) 2
0.075% Bisulfite 0.2 2.6 2.1 7.7 (CD:Arg 1.0:1.5) 3 0.075%
Bisulfite 0.25 3.1 2.1 9.8 (CD:Arg 1.0:1.2)
[0118] The results as presented in Tables M and N indicate that
formulations containing more arginine are more stable when exposed
to air at 25.degree. C. (formulations 2 vs. 3). Further,
formulations containing Na-bisulfate are less stable than the
formulation containing ascorbic acid and L-cysteine (formulations 2
vs. 1, respectively) when stored under nitrogen (anaerobic
conditions). N.sub.2 provided significant protection from
degradation and degradant formation. Formulations exposed to air
are more stable when kept refrigerated, as compared to room
(ambient) temperature.
Example 10: Effect of Various Antioxidants on the Stability of
Formulations Containing 4% Carbidopa at 40.degree. C.
[0119] Liquid formulations with carbidopa and arginine (Table O)
were prepared as described above. The effect various antioxidants
on the stability of formulations containing 4% CD at 40.degree. C.
was evaluated using HPLC analysis. The levels of the degradant and
total impurities, presented in tables P and Q respectively,
indicate the level of stability of carbidopa formulations.
TABLE-US-00026 TABLE O 1 2 3 4 Carbidopa 4.0 4.0 4.0 4.0 L-Arginine
3.4 3.7 4.6 4.6 N-MP 3.5 0 0 0 Ascorbic acid 0 0 0.4 0.5 Cysteine 0
0 0.2 0.1 Na bisulfite 0.1 0.075 0 0 Total solutes 12.9 9.2 9.2 9.2
Water 87.1 90.8 90.8 90.8 L-Arg (mM) 195 212 265 265 CD (mM) 177
177 177 177 Ratio L-Arg/CD 1.1 1.2 1.5 1.5 pH measured 8.76 8.96
9.13 9.12
TABLE-US-00027 TABLE P Degradant t = 2 d/ t = 5 d/ t = 0 40.degree.
C. 40.degree. C. 1 4% CD - 3.4% L-Arg + 0.27 .+-. 0.01 1.52 .+-.
0.56 2.60 .+-. 0.67 3.5% N-MP 0.1% sodium bisulfite 2 4% CD - 3.7%
L-Arg 0.30 .+-. 0.01 1.41 .+-. 0.54 2.95 .+-. 0.60 0.075% sodium
bisulfite 3 4% CD - 4.62% L-Arg 0.06 .+-. 0.01 0.38 .+-. 0.14 1.26
.+-. 0.45 0.4% ascorbic + 0.2% L-cysteine 4 4% CD - 4.62% L-Arg
0.11 .+-. 0.01 0.50 .+-. 0.23 1.97 .+-. 0.52 0.5% ascorbic + 0.1%
L-cysteine
TABLE-US-00028 TABLE Q Total impurities t = 2 d/ t = 5 d/ t = 0
40.degree. C. 40.degree. C. 1 4% CD - 3.4% L-Arg + 3.5% 0.61 2.00
.+-. 0.54 3.19 .+-. 0.65 N-MP 0.1% sodium bisulfite 2 4% CD - 3.7%
L-Arg 0.60 1.86 .+-. 0.56 3.61 .+-. 0.60 0.075% sodium bisulfite 3
4% CD - 4.62% L-Arg 0.29 0.92 .+-. 0.16 2.15 .+-. 0.59 0.4%
ascorbic + 0.2% L-cysteine 4 4% CD - 4.62% L-Arg 0.34 1.17 .+-.
0.28 2.82 .+-. 0.61 0.5% ascorbic + 0.1% L-cysteine
[0120] The results presented in Tables P and Q suggest that
formulations containing Na-bisulfite are less stable than the
formulations containing ascorbic acid and L-cysteine (formulations
1 & 2 vs. 3 & 4), both during preparation and when stored
at 40.degree. C. Furthermore, there was a cysteine dose-response,
i.e., the higher the concentration of L-cysteine, the less
degradant was formed. No dose response was observed with
Na-bisulfite, suggesting that the maximum possible protection may
be attained with 0.075% Na-bisulfite.
Example 11: Effect of Ascorbic Acid Combined With Various
Antioxidants on the Stability of Formulations Containing 4%
Carbidopa
[0121] Liquid formulations with carbidopa and ascorbic acid with or
without additional antioxidants were prepared as described above.
The combination effect between ascorbic acid and various
antioxidants on the stability of formulations containing 4% CD at
25.degree. C. was evaluated using HPLC analysis. The levels of the
degradant and total impurities presented in tables R and S
respectively, point toward the level of stability of carbidopa
formulations.
TABLE-US-00029 TABLE R Degradant t = 3 d at t = 2 d at 4% CD t = 0
25.degree. C./N.sub.2 25.degree. C./O.sub.2 Without antioxidants
0.24 1.29 3.27 0.5% Asc 0.54 3.17 5.09 0.5% Asc + 0.2% bisulfite
0.16 0.76 2.98 0.5% Asc + 0.2% cysteine 0.09 0.27 2.67 0.5% Asc +
0.2% NAC 0.14 0.83 3.64 0.75% bisulfite (CONTROL) 0.39 1.23
2.98
TABLE-US-00030 TABLE S Total impurities t = 3 d at t = 2 d at 4% CD
t = 0 25.degree. C./N.sub.2 25.degree. C./O.sub.2 Without
antioxidants 1.04 1.88 4.01 0.5% Asc 1.07 3.96 6.40 0.5% Asc + 0.2%
bisulfite 0.70 1.29 3.64 0.5% Asc + 0.2% cysteine 0.67 0.74 3.43
0.5% Asc + 0.2% NAC 0.72 1.36 4.49 0.75% bisulfite (CONTROL) 1.13
1.71 3.58
[0122] Results presented in Tables R and S show that ascorbic acid,
0.5%, was insufficient for the prevention of degradant formation in
a formulation containing carbidopa. Furthermore, ascorbic acid
requires another antioxidant in order to exert its maximum
antioxidant activity, for example ascorbic acid, 0.5%, and
L-cysteine, NAC, or Na-bisulfate inhibited carbidopa degradation in
a synergistic manner. Formulations containing ascorbic acid and
L-cysteine had the lowest amount of degradant after 3 days at
25.degree. C.
[0123] The effect of Na-bisulfate on carbidopa degradation was
similar to that obtained with no antioxidants at all.
Example 12: Effect of Antioxidants on the Stability of 4% CD
Formulations Stored at 25.degree. C.
[0124] Liquid formulations with carbidopa and arginine (Table T)
were prepared as described above. The effect of various
antioxidants on the stability of formulations containing 4%
carbidopa at 25.degree. C. was evaluated using HPLC analysis. The
levels of the degradant and total impurities presented in Table W,
point toward the level of stability of carbidopa formulations.
TABLE-US-00031 TABLE T 1 2 3 4 5 6 7 Carbidopa 4.0 4.0 4.0 4.0 4.0
4.0 4.0 L-Arginine 4.6 4.6 4.6 4.6 4.6 4.6 4.6 Ascorbic acid 0.2
0.5 0.2 0.0 0.0 0.0 0.2 Cysteine 0.0 0.2 0.2 0.2 0.2 0.0 0.0 Na
bisulfite 0.0 0.0 0.0 0.0 0.1 0.1 0.1
TABLE-US-00032 TABLE W Degradant TOTAL Impurities t = 0 t = 1 wk t
= 0 t = 1 wk 1 0.2% ascorbic acid 0.48 1.43 1.20 2.11 2 0.5%
ascorbic acid/0.2% 0.12 0.17 0.81 0.72 cysteine 3 0.2% ascorbic
acid/0.2% 0.12 0.21 0.85 0.94 cysteine 5 0.2% cysteine 0.26 0.53
1.02 1.00 5 0.2% cysteine/0.1% 0.71 1.01 1.62 1.66 bisulfite 6 0.1%
bisulfite 0.33 0.84 1.16 1.45 7 0.2% ascorbic acid/0.1% 0.26 0.70
0.99 1.49 bisulfite
[0125] Results presented in Table W suggest that ascorbic acid,
bisulfite, or cysteine, each used alone, did not inhibit degradant
formation. Combinations between bisulfite and cysteine or ascorbic
acid did not inhibit degradant formation. There was a synergistic
inhibitory effect on degradant formation between ascorbic acid and
cysteine, but no such synergism between cysteine and bisulfite was
observed. Such synergistic effects can be seen between ascorbic
acid and bisulfite (with higher ascorbic acid concentrations).
These results further suggest that formulations containing the
unique combination of ascorbic acid and cysteine may provide the
best means for the inhibition of degradant formation.
[0126] Ascorbic acid, at 0.2%, was not sufficient for the
prevention of degradant formation. With 0.2% cysteine, 0.5%
ascorbic acid was more effective than 0.2% in reducing the total
amount of impurities and degradant formation, suggesting that at
least 0.5% ascorbic acid with 0.2% cysteine is desirable.
Example 13: Determination of the Level of Hydrazine in Carbidopa
Formulations
[0127] The determination of hydrazine was carried out by
derivatization using Aceton-d6. The hydrazine derivative was
analyzed by gas chromatography mass spectrometry (GC/MS). The
specific mass of hydrazine derivative was measured in the selected
ion monitoring mode (SIM-mode) according to Solvias standard
operating procedures (SOP's).
[0128] Liquid formulations with carbidopa, levodopa, and arginine
(Table X) were prepared as described above. The levels of hydrazine
in the referenced formulations were measured (Table Y).
TABLE-US-00033 TABLE X Formulation Composition (%) Formulation # 1
2 3 4 5 6 7 8 9 10 Carbidopa 4 4 1.4 1.4 1.4 1.4 0.75 1.4 0.75 0.75
Levodopa -- -- 6 6 6 6 6 6 6 6 Arginine 4.6 4.6 14.8 15.5 15.5 15.5
15.2 15.5 15.2 15.2 Ascorbic 0.4 0.5 0.75 0.5 0.5 0.5 0.5 0.5 0.5
0.5 acid L-Cysteine 0.2 0.1 -- 0.4 0.4 -- -- 0.4 0.4 0.4 NAC -- --
-- -- -- 0.5 0.5 -- -- -- Tween 80 -- -- -- -- -- -- -- -- -- 0.3
Sodium -- -- -- -- -- -- -- -- -- -- Ascorbate Formulation # 11 12
13 14 15 16* 17** 18 19 20 Carbidopa 1.4 0.75 1.4 1.4 3 1.5 1.5 1.4
3 1.3 Levodopa 6 6 6 6 12 6 6 6 12 12 Arginine 15.5 15.2 15.5 15.5
32.3 15.5 15.5 15.5 32.3 29 Ascorbic 0.5 0.5 0.5 0.5 -- 0.5 0.5 0.5
acid L-Cysteine 0.4 -- -- 0.4 0.3 0.4 0.4 0.4 NAC -- 0.5 0.5 -- --
-- -- -- 0.3 0.3 Tween 80 0.3 0.3 0.3 0.3 -- 0.3 0.3 0.3 Sodium --
-- -- -- 1.2 -- -- -- 1.2 1.2 Ascorbate *Sealed with N2 **Sealed
with O.sub.2
TABLE-US-00034 TABLE Y Time and temp prior to Storage in analysis
(in- Hydrazine (ppm) vials use stability) 1 2 3 4 5 6 7 8 9 10 T =
0 at -20.degree. C. 1 month 24 h 25.degree. C. 0.82 0.77 0.39 at
-20.degree. C. 48 h 0.73 0.71 0.43 24 h 37.degree. C. 0.76 0.84
0.41 48 h 0.87 0.93 0.59 3 months <0.1 <0.1 at -20.degree. C.
6 months 3 free-thaw 0.1 0.1 at -20.degree. C. cycles (25.degree.
C.) 0.1 9 months 0.1 at -20.degree. C. 1 year 24 h 25.degree. C.
<0.1 at -20.degree. C. 37.degree. C. <0.1 0.1 24 hr
37.degree. C. 7 days <0.1 <0.1 <0.1 at 25.degree. C. 24 h
37.degree. C. 0.1 <0.1 1 month 25.degree. C. Time and temp prior
to Storage in analysis (in- Hydrazine (ppm) vials use stability) 11
12 13 14 15 16 17 18 19 20 T = 0 <0.1 <0.1 <0.1 at
-20.degree. C. 1 month 24 h 25.degree. C. at -20.degree. C. 48 h 24
h 37.degree. C. 48 h 3 months 0.1 0.1 at -20.degree. C. 6 months 3
free-thaw at -20.degree. C. cycles (25.degree. C.) 0.1 <0.1 0.1
0.1 9 months 0.1 0.1 at -20.degree. C. 1 year 24 h 25.degree. C. at
-20.degree. C. 37.degree. C. 0.1 <0.1 0.1 0.1 24 hr 37.degree.
C. <0.1 0.1 <0.1 7 days at 25.degree. C. 24 h 37.degree. C. 1
month 0.1 0.3 0.1 25.degree. C.
[0129] The results presented in Table Y clearly show that the
levels of hydrazine is at least 2 fold lower in levodopa
formulations versus formulations without levodopa. Furthermore,
formulations comprising L-cysteine or NAC show at least 4-fold
lower levels of hydrazine compared to formulations without
L-cysteine or NAC.
Example 14: Carbidopa/Levodopa Formulations
[0130] Based on the discoveries of combinations that have reduced
degradant and hydrazine formation, we have developed new CD/LD
formulations. These formulations are shown in Tables Z and AA
below.
TABLE-US-00035 TABLE Z DS Ascorbic L- Tween- (%) LD CD Arginine
Acid Cysteine NAC 80 pH 1 6 1.4 15.5 0.5 0.4 -- 0.3 9.4-9.6 2 6 1.4
15.5 0.5 -- 0.5 0.3 9.4-9.6 3 6 0.75 15.2 0.5 0.4 -- 0.3 9.4-9.6 4
6 0.75 15.2 0.5 -- 0.5 0.3 9.4-9.6 Margins 6 0.6-1.4 15-16 0.5 0.4
0.5 0.3 9.4-9.6
TABLE-US-00036 TABLE AA DS Sodium L- Cysteine- Tween- (%) LD CD
Arginine Meglumine Ascorbate Cysteine NAC HCl 80 pH 1 12 3 32 --
1.2 0.3 -- -- -- 9.6-9.8 2 13.2 3.3 36 -- 1.3 0.3 -- -- -- 9.6-9.8
3 13.2 3.3 -- 36 1.3 0.3 -- -- -- 9.6-9.8 4 12 3 -- 32 1.2 -- 0.3
-- -- 9.6-9.8 5 12 3 32 -- 1.2 -- 0.3 -- -- 9.6-9.8 12-15 1.2-4
32-42 32-42 1.0-1.3 0.1-0.5 .ltoreq.2 9.6-9.8 * ** * Can replace
L-cysteine. ** Optionally added to stabilize the formulation.
[0131] Additional formulations that may be used in the context of
those disclosed herein are provided in Table BB below. The
formulations may include additional components (e.g., any of those
described herein). Tables CC and DD described further formulations
that can be used in the context of those described herein.
TABLE-US-00037 TABLE BB Levodopa Carbidopa Amino Acid Other Conc.
(%) Conc. (%) Name Conc. (%) Name Conc. (%) 3 0 Lys 5.6 2.5 0 Lys
4.6 1.25 0 His 2.5 9.5 0 Arg 15.9 4.8 1.4 Arg 11.0 4.8 1.4 Arg 12.1
4.8 1.4 Arg 12.7 5.4 1.5 Arg 13.5 5.4 1.5 Arg 14.8 6 1.5 Arg 14.8 6
1.5 Arg 16.0 7 2 Arg 17.8 7 1.5 Arg 14.1 Dextrose 5.0 8 1.5 Arg
15.7 Dextrose 5.0 10 1.5 Arg 19.2 Dextrose 5.0 6 1.5 Arg 9.3 NaOH
4.6 5 0 Meglumine 10.8 8 1.5 Arg 15.7 Meglumine 3.2 8 1.5 Arg 12.2
Meglumine 7.9 10 1.5 Arg 19.2 Meglumine 4.0 10 1.5 Arg 14.6
Meglumine 9.9 7 1.5 Arg 14.1 Meglumine 2.8 7 1.5 Arg 10.7 Meglumine
6.9 6 1.5 Arg 13.5 Na-Asc 1 6 1.5 Arg 14.2 Na-Asc 1 6 1.5 Arg 14.8
Na-Asc 1 6 1.5 Arg 16.0 Na-Asc 1 4.8 1.4 Arg 11.0 Na-Asc 1 4.8 1.4
Arg 11.6 Na-Asc 1 4.8 1.4 Arg 12.1 Na-Asc 1 4.8 1.4 Arg 12.7 Na-Asc
1 6 1.5 Arg 14.8 Asc 1 6 1.5 Arg 15.8 Na-Asc 1 6 1.5 Arg 15.8 Asc 1
6 1.5 Arg 16.8 Na-Asc 1 6 1.5 Arg 16.8 Asc 1 5.4 1.5 Arg 12.3
Na-Asc 1 5.4 1.5 Arg 12.3 Asc 1 5.4 1.5 Arg 13.5 Na-Asc 1 5.4 1.5
Arg 13.5 Asc 1 5.4 1.5 Arg 14.8 Na-Asc 1 5.4 1.5 Arg 14.8 Asc 1 6
1.5 Arg 16.0 Asc 1 7 2 Arg 17.8 Asc 1 7 2 Arg 17.8 Na-Asc 1 12 3
Arg 24.4 12 3 Arg 29.6 12 3 Arg 32.1 7 0 Arg 7 0.5 Arg 7 1 Arg 7
1.5 Arg 7 2 Arg 6 0 Arg 13.5 6 0.5 Arg 14.2 6 1 Arg 14.8 6 2 Arg
16.5 0 2
TABLE-US-00038 TABLE CC Property 1 2 3 API Concentration 2 & 4%
4% 0.6-20% CD:Arginine ratio 1:1.1-1.2 1:1.5 1:.gtoreq.1 Exipients
NMP 3.5% 0 0-15% concentra- Na-bisulfite 0.1% 0 0-0.2% tion
Ascorbic Acid 0 0.75% 0-2% or more L-Cysteine 0 0.1% 0-0.5% or more
or NAC Other anti- -- -- 0-2% oxidants Osmolality 650-750 300-400
200-1800 for SC No limits for ID pH 8.2-8.6 8.6-9.1 8-9.8 Stability
25.degree. C. 48-72 hrs .gtoreq.21 d 2 wks-.gtoreq.2 yrs 4.degree.
C. Not stable .gtoreq.21 d 2 wks-.gtoreq.2 yrs -20.degree. C.
.gtoreq.1 year .gtoreq.21 d 2 wks-.gtoreq.2 yrs SC Infusion/24 hrs
2 ml 2 ml 0.1-20 ml
TABLE-US-00039 TABLE DD Property 4 5 6 API CD 0 or 1 1-2% 0-4% up
Concen- or 2% to 6% tration LD 3-7% 5-7% 2.5-12% up to 14% Ratios
LD to CD 6:1-6:3 3.5-4:1 1:1-10:0.5 Ratio or LD alone CD:Arginine
1:1.2 1:9-14 1:.gtoreq.35 Ratio LD:Arginine 1:1.8-2.2 1:2-3.5
1:.gtoreq.3.6 Ratio API:Argi- [1:1.2 1:2.3-2.5 1:.gtoreq.1.8 nine
CD:Arg + 1:2 Ratio LD:Arg] + 12.5% Arg Exepients NMP 0 0 0
Na-bisulfite 0.075-0.15% 0 0-0.2% Ascorbic 0 0.75 0-2% or more Acid
Other anti- -- -- 0-2% oxidants Osmolal- 9/1% 1300-1500 -- 200-1800
for ity LD/CD SC 7/2% 950-1150 1200-1300 No limits LD/CD for ID
6/1.5% 800-850 940-980 LD/CD 5/1.25% NT 790-830 LD/CD pH 8.5-9.5
9.2-9.6 9.1-9.8 Stability 25.degree. C. .gtoreq.2 days .gtoreq.2
days .gtoreq.2 days 4.degree. C. <2 days .gtoreq.2 days
.gtoreq.2 days -20.degree. C. .gtoreq.2 days .gtoreq.2 days
.gtoreq.2 days SC Infusion/24 hrs 2 ml 2-6 ml 0.1-10 ml/site
Intraduodenal/24 hrs -- -- 4-24 ml Intrathecal -- -- 1-1000
.mu.l/day
EQUIVALENTS
[0132] While specific embodiments of the subject invention have
been discussed, the above specification is illustrative and not
restrictive. Many variations of the invention will become apparent
to those skilled in the art upon review of this specification. The
full scope of the invention should be determined by reference to
the claims, along with their full scope of equivalents, and the
specification, along with such variations.
[0133] Unless otherwise indicated, all numbers expressing
quantities of ingredients, reaction conditions, and so forth used
in the specification and claims are to be understood as being
modified in all instances by the term "about." Accordingly, unless
indicated to the contrary, the numerical parameters set forth in
this specification and attached claims are approximations that may
vary depending upon the desired properties sought to be obtained by
the present invention.
INCORPORATION BY REFERENCE
[0134] The entire contents of all patents, published patent
applications, websites, and other references cited herein are
hereby expressly incorporated herein in their entireties by
reference.
* * * * *