U.S. patent application number 15/834514 was filed with the patent office on 2018-04-12 for composition and method of use of colchicine oral liquid.
This patent application is currently assigned to RxOMEG Therapeutics LLC. The applicant listed for this patent is RxOMEG Therapeutics LLC. Invention is credited to Indu Muni, Naomi Vishnupad.
Application Number | 20180098938 15/834514 |
Document ID | / |
Family ID | 59787983 |
Filed Date | 2018-04-12 |
United States Patent
Application |
20180098938 |
Kind Code |
A1 |
Muni; Indu ; et al. |
April 12, 2018 |
COMPOSITION AND METHOD OF USE OF COLCHICINE ORAL LIQUID
Abstract
Oral liquid colchicine formulations are described herein.
Methods of using the oral liquid colchicine formulations are also
provided.
Inventors: |
Muni; Indu; (North Reading,
MA) ; Vishnupad; Naomi; (Reading, MA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
RxOMEG Therapeutics LLC |
Woburn |
MA |
US |
|
|
Assignee: |
RxOMEG Therapeutics LLC
Woburn
MA
|
Family ID: |
59787983 |
Appl. No.: |
15/834514 |
Filed: |
December 7, 2017 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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15818905 |
Nov 21, 2017 |
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15834514 |
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15358621 |
Nov 22, 2016 |
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15818905 |
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62306232 |
Mar 10, 2016 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 47/02 20130101;
A61K 31/165 20130101; A61K 9/0095 20130101; A61K 47/36 20130101;
A61K 47/12 20130101; A61K 47/10 20130101 |
International
Class: |
A61K 9/08 20060101
A61K009/08; A61K 9/00 20060101 A61K009/00; A61K 31/165 20060101
A61K031/165; A61K 47/26 20060101 A61K047/26; A61K 47/24 20060101
A61K047/24; A61K 47/10 20060101 A61K047/10; A61K 47/36 20060101
A61K047/36; A61K 47/12 20060101 A61K047/12 |
Claims
1. A liquid solution suitable for oral administration comprising
colchicine, wherein the concentration of colchicine in the solution
is 0.01-1.0 mg/mL, and a pharmaceutically acceptable solvent
system, and wherein the liquid solution has a stable pH and is
stable for 30 days to 24 months at ambient and refrigerated
temperature conditions.
2. The liquid solution of claim 1, wherein the liquid solution
comprises a glycol, buffering agent, preservative, sweetener, and
thickening agent.
3. The liquid solution of claim 2, wherein the liquid solution is
comprised of the following components: benzyl alcohol citric acid
colchicine dibasic sodium phosphate heptahydrate propylene glycol
glycerin xanthan gum, and water.
4. The liquid solution of claim 1, wherein the liquid solution has
a bioavailability in vivo similar to single ingredient colchicine
tablets and capsules.
5. The liquid solution of claim 1, wherein the liquid solution has
a viscosity in the range of 40-800 cps and is stable for 30 days to
24 months at ambient and refrigerated temperature conditions.
6. A liquid solution suitable for oral administration comprising
colchicine and a pharmaceutically acceptable solvent system,
wherein the liquid solution is stable for 30 days to 24 months at
ambient and refrigerated temperature conditions, and wherein the
liquid solution comprises a glycol, buffering agent, preservative,
sweetener, and thickening agent.
7. The liquid solution of claim 6, wherein the liquid solution is
comprised of the following components: benzyl alcohol citric acid
colchicine propylene glycol glycerin, and water.
8. The liquid solution of claim 6, wherein the solution is stored
in a light-resistant container.
9. The liquid solution of claim 6, wherein the liquid solution has
a viscosity in the range of 40-800 cps and a stable pH and is
stable for 30 days to 24 months at ambient and refrigerated
temperature conditions.
10. A liquid solution suitable for oral administration comprising
colchicine, wherein the concentration of colchicine in the solution
is 0.01-1.0 mg/mL, and a pharmaceutically acceptable solvent
system, wherein the liquid solution is stable for 30 days to 24
months at ambient and refrigerated temperature conditions, and
wherein the liquid solution has a bioavailability in vivo similar
to single ingredient colchicine tablets and capsules.
11. The liquid solution of claim 10, wherein the liquid solution
comprises water, 0.1-0.3% w/v of buffering agents, 2-18% glycols,
0.2-0.4% w/v of preservatives, and 0.1-0.2% w/v of thickening
agents.
12. The liquid solution of claim 11, wherein the solution has less
than 0.5% of degradants and wherein the degradants comprise
.beta.-lumicolchicine, .gamma.-lumicolchicine, and/or
colchiceine.
13. The liquid solution of claim 10, wherein the liquid solution
has a bioavailability in vivo similar to single ingredient
colchicine tablets and capsules.
14. The liquid solution of claim 10, wherein the liquid solution
has a stable pH.
15. A liquid solution suitable for oral administration comprising
colchicine, wherein the concentration of colchicine in the solution
is 0.01-1.0 mg/mL, and a pharmaceutically acceptable solvent
system, and wherein the liquid solution is stable for 30 days to 24
months at ambient and refrigerated temperature conditions and
wherein the solution has less than 0.5% of degradants, optionally
wherein the degradants comprise .beta.-lumicolchicine,
.gamma.-lumicolchicine, and/or colchiceine.
16. The liquid solution of claim 15, wherein the liquid solution
comprises a glycerin.
17. The liquid solution of claim 15, wherein the liquid solution
comprises water, 0.1-0.3% w/v of buffering agents, 2-18% glycols,
0.2-0.4% w/v of preservatives, and 0.1-0.2% w/v of thickening
agents.
18. The liquid solution of claim 16, wherein the liquid solution
has a viscosity in the range of 40-800 cps.
19. The liquid solution of claim 16, wherein the liquid solution
prevents photodegradation of colchicine.
20. The liquid solution of claim 16, wherein the solution is stored
in a light-resistant container.
Description
RELATED APPLICATIONS
[0001] This application is a continuation of U.S. application Ser.
No. 15/818,905, filed on Nov. 21, 2017, entitled "COMPOSITION AND
METHOD OF COLCHICINE ORAL LIQUID" which is a divisional of U.S.
application Ser. No. 15/358,621, filed on Nov. 22, 2016, entitled
"COMPOSITION AND METHOD OF USE OF COLCHICINE ORAL LIQUID" which
claims the benefit under 35 U.S.C. .sctn. 119 of U.S. provisional
application 62/306,232, filed Mar. 10, 2016, the entire contents of
which is incorporated herein by reference.
BACKGROUND OF THE INVENTION
[0002] Colchicine is an alkaloid compound found in plant extracts
that is used to treat gout, familial Mediterranean fever (FMF),
pericarditis, Behcet's disease, and atrial fibrillation. Colchicine
has also been used to treat amyloidosis, calcium pyrophosphate
deposition disease (pseudogout), cirrhosis of the liver, sarcoid
arthritis, and inflammatory diseases. Colchicine is administered to
patients as a solid oral dosage form, such as a tablet or capsule.
A standard dosage of colchicine is typically administered to
patients once or twice a day.
[0003] Colchicine has previously been shown to be unstable at room
temperature in solution. An article published in Drug Development
and Industrial Pharmacy, 15(11), 1905-1909 (1989) by Habib, et.
al., investigated the stability of colchicine and showed that there
is photodegradation of colchicine in solution, especially in the
presence of glycerin. Other additives, such as lithium carbonate,
p-aminobenzoic acid, and uric acid, used in this study did not
prevent the degradation of the colchicine, and furthermore, are not
acceptable excipients for an oral solution.
SUMMARY OF THE INVENTION
[0004] The invention in some aspects is a method of treating a
colchicine sensitive disorder, by orally administering a
composition, comprising an oral liquid colchicine formulation to a
human subject having a colchicine sensitive disorder in an
effective amount to treat the disorder. The colchicine sensitive
disorder in some embodiments is selected from gout, familial
Mediterranean fever (FMF), pericarditis, Behcet's disease, atrial
fibrillation, amyloidosis, calcium pyrophosphate deposition disease
(pseudogout), cirrhosis of the liver, sarcoid arthritis,
inflammatory diseases, and Disk diseases & related spinal
disorders. In some embodiments the oral liquid colchicine
formulation is any of the oral liquid colchicine formulations
described herein.
[0005] The invention in other aspects is a pharmaceutical solution
or suspension suitable for oral administration comprising
colchicine and a pharmaceutically acceptable solvent system
comprised of one or more agents selected from the group consisting
of water, glycols, buffering agents, sweeteners, flavoring agents,
preservatives and dyes. The buffering agent in some embodiments is
about 0.12% (w/v) citric acid (anhydrous). In some embodiments the
sweetener is about 0.2% (w/v) sucralose. In other embodiments the
flavoring agent is about 0.125% (w/v) artificial grape flavor
and/or Flavor Cherry 825.662. In some embodiments the preservative
is about 0.3% (w/v) of benzyl alcohol. In some embodiments the
preservative is about 0.2% (w/v) of citric acid, anhydrous. In
other embodiments the dye comprises one or more of D&C Yellow
No. 10 and FD&C Red No. 40. In some embodiments the dye
comprises about 0.01% (w/v) FD&C Red No. 40. In some
embodiments the thickening agent comprises about 0.15% (w/v)
xanthan gum. The concentration of colchicine in the solution in
other embodiments is 0.01-1.0 mg/ml or 0.2-0.60 mg/ml.
[0006] In other embodiments the pharmaceutically acceptable solvent
system is comprised of one or more agents selected from the group
consisting of water, propylene glycol, glycerin, benzyl alcohol,
parabens, citric acid, xanthan gum, sucralose, a dye, and a
flavoring agent and/or taste enhancing agent.
[0007] In yet other embodiments the pharmaceutically acceptable
solvent system is comprised of the following components:
TABLE-US-00001 Ingredient % w/v Benzyl Alcohol 0.3 Citric Acid,
Anhydrous 0.2 Colchicine* 0.012 FD&C Red No. 40 0.01 Dibasic
Sodium Phosphate, Heptahydrate 1.2 Flavor Cherry 825.662 0.125
Propylene Glycol 5 Glycerin 5 Sucralose 0.15 Xanthan Gum 0.15 Water
Q.S. *calculated on the anhydrous, solvent free basis
[0008] In some embodiments the oral liquid formulations described
herein are non-sterile.
[0009] In some embodiments the oral liquid formulations comprises
compatible and stable preservative. In other embodiments the
compatible and stable preservative is benzyl alcohol.
[0010] In some embodiments the colchicine formulation has a
viscosity in the range of 40-800 cps. In other embodiments the
colchicine formulation has a viscosity of 80-250 cps.
[0011] In other embodiments the colchicine formulation is volume
packaged 60 mL-473 mL, and more preferably 150 ml-300 mL.
[0012] The colchicine formulations of the invention may be provided
in any type of bottle acceptable for oral liquid medications. In
some embodiments the colchicine formulation is packaged in a
plastic bottle, such as HDPE (high density polyethylene bottles) or
PET. The colchicine solution may also be packaged in glass bottles,
which could be amber or clear, in other embodiments. The caps of
the bottles may or may not have an induction seal and could be easy
to open or tamper resistant. The bottles and/or the packaging could
be multi use or single dose units. A preferred embodiment is a 190
ml HDPE container closure system with a cap with an induction
seal.
[0013] In some embodiments the oral liquid colchicine formulation
is stable at room temperature for at least 3 months, at least 6
months, at least 18 months, or at least 24 months. In some
embodiments the oral liquid colchicine formulation is stable at
accelerated temperatures for at least 1 month, at least 2 months,
at least 3 months, or at least 6 months. In some embodiments the
oral liquid colchicine formulation is determined to be stable when
the solution has less than 5%, less than 4%, less than 3%, less
than 2%, less than 1% or less than 0.5% of any one degradant. In
other embodiments the oral liquid colchicine formulation is
determined to be stable when the solution has less than 5%, less
than 4%, less than 3%, less than 2%, less than 1% or less than 0.5%
of total degradants. In some embodiments, degradants include .beta.
lumicolchicine, .gamma.-lumicolchicine, colchiceine, and any other
individual unknown impurities.
[0014] Each of the limitations of the invention can encompass
various embodiments of the invention. It is, therefore, anticipated
that each of the limitations of the invention involving any one
element or combinations of elements can be included in each aspect
of the invention. This invention is not limited in its application
to the details of construction and the arrangement of components
set forth in the following description or illustrated in the
drawings. The invention is capable of other embodiments and of
being practiced or of being carried out in various ways. Also, the
phraseology and terminology used herein is for the purpose of
description and should not be regarded as limiting. The use of
"including," "comprising," or "having," "containing", "involving",
and variations thereof herein, is meant to encompass the items
listed thereafter and equivalents thereof as well as additional
items.
BRIEF DESCRIPTION OF DRAWINGS
[0015] The figures are illustrative only and are not required for
enablement of the invention disclosed herein.
[0016] FIG. 1 depicts a chromatograph. Sample, colchicine standard,
and diluent blank can be found from top to bottom.
[0017] FIG. 2 depicts a chromatograph of an exemplary liquid oral
colchicine solution. Diluent blank, placebo, red color, benzyl
alcohol, flavor cherry, and colchicine standard are shown from top
to bottom.
[0018] FIG. 3 shows the calibration curve of colchicine in Caco-2
transport buffer.
[0019] FIG. 4 shows a chromatogram of colchicine in Caco-2
buffer.
[0020] FIGS. 5A-5F show representative colchicine chromatograms.
FIGS. 5A-5B show chromatograms of liquid colchicine dosing (FIG.
5A) and liquid A.fwdarw.B receiver sample (FIG. 5B). FIGS. 5C-5D
show chromatograms of colchicine USP tablet dosing (FIG. 5C) and
liquid A.fwdarw.B receiver sample (FIG. 5D). FIGS. 5E-5F show
chromatograms of colchicine capsule dosing (FIG. 5E) and liquid
A.fwdarw.B receiver sample (FIG. 5F).
DETAILED DESCRIPTION
[0021] Colchicine is administered to patients as a solid oral
dosage form, such as a tablet or capsule. Colchicine has previously
been shown to be unstable at room temperature in solution. Habib et
al. showed rapid photodegradation of colchicine in solution,
especially in the presence of glycerin. Other additives, such as
lithium carbonate, p-aminobenzoic acid, and uric acid, were used in
this study, but did not prevent the degradation of the colchicine,
and furthermore, are not acceptable excipients for an oral
solution. Surprisingly, it was found according to the invention,
that liquid suspensions or solutions of colchicine formulated as an
oral solution are stable at ambient temperature and have stable pH
for extended periods of time. For instance the liquid solutions or
suspensions described herein are stable for at least three months
in refrigerated, ambient, and accelerated temperatures. The
findings of the invention have important clinical implications. An
oral solution or suspension of colchicine is advantageous for
colchicine dosing and administration.
[0022] Colchicine,
(-)-N-[(7S,12aS)-1,2,3,10-tetramethoxy-9-oxo-5,6,7,9-tetrahydrobenzo[a]he-
ptalen-7-yl]-acetamide, is a pale yellow powder soluble in water in
1:25 dilution. Colchicine is an alkaloid found in extracts of
certain plants such as Colchicum autumnale and Gloriosa superba.
Colchicine arrests cell division in animals and plants. It has
adversely affected spermatogenesis in humans and in some animal
species under certain conditions.
[0023] The invention encompasses liquid formulations of colchicine.
The present invention provides for liquid formulation of
colchicine, suitable for oral administration that is stable at room
temperature. The liquid formulation can be either a solution or a
suspension. Colchicine solid oral dosage forms, such as tablets and
capsules have been used for the prophylactic treatment of gout and
to treat patients suffering from gout flares. In addition to
treating patients with gout, colchicine is also used to treat
patients with Familial Mediterranean Fever (FMF). Studies have also
shown that colchicine may be used to treat patients with
cardiovascular disease and various other conditions.
[0024] Currently colchicine is primarily used to treat patients
suffering from gout. An oral liquid formulation can provide
physicians more flexibility in designing dosage regimens for their
patients. This is particularly important since colchicine is toxic
and has a narrow therapeutic index. The methods described herein
are useful for the treatment of gout. The treatment of gout
involves the prophylactic treatment of gout as well as the
treatment of gout flares. The prophylactic treatment of gout refers
to the treatment of a patient who has had one or more gout flares,
in order to reduce the occurrence of future gout flares.
[0025] Some of the challenges in formulating an oral liquid of
colchicine include maintaining stability of the colchicine,
maintaining an optimum pH, and masking the bitter taste. It is also
important to establish an effective preservative system to prevent
the growth of bacteria, mold, and other contaminants. Additionally,
the oral liquid colchicine must be patient friendly and requires
suitable packaging, such as a container closure system that factors
in the potential effects of light and air exposure.
[0026] Also provided herein are methods of treating gout, familial
Mediterranean fever (FMF), Behcet's disease, cardiovascular disease
(atrial fibrillation, pericarditis), amyloidosis, calcium
pyrophosphate deposition disease (pseudogout), cirrhosis of the
liver, sarcoid arthritis, inflammatory diseases, and Disk diseases
& related spinal disorders comprising administering to a
patient, such as a child or an elderly patient, an oral liquid
formulation compounded from colchicine as described herein. In some
embodiments, oral liquid formulations disclosed herein can also be
used to treat for other conditions (e.g., skin conditions) known in
the art (Ben-Chetrit E, Levy M. Colchicine: 1998 update. Semin
Arthritis Rheum. 1998; Yurdakul S, Mat C, Tuzun Y, Ozyazgan Y,
Hamuryudan V, Uysal O, Senocak M, Yazici H. A double-blind trial of
colchicine in Behcet's syndrome. Arthritis Rheum. 2001 November;
44(11):2686-92. August; 28(1):48-59; Molad Y. Update on colchicine
and its mechanism of action. Curr Rheumatol Rep. 2002 June;
4(3):252-6).
[0027] Commonly, geriatric populations encounter difficulty being
administered solid oral dosage forms such as tablets and capsules.
This may lead to non-compliance with the recommended
pharmacotherapy with the solid oral dosage forms and likely results
in rendering the therapy ineffective. Solid oral dosage forms are
usually not favorable for geriatric populations due to the
potential risk of choking. Additionally, certain solid oral dosage
forms of medications cannot be administered simply by crushing
(e.g., patients requiring various types of feeding tubes) because
of the coating or drug delivery mechanism by which the drug is
released.
[0028] As used herein, "colchicine" refers to colchicine base, its
salt, or solvate or derivative or isomer or polymorph thereof.
Suitable compounds include the free base, the organic or inorganic
salts, isomers, isomer salts, solvates, polymorphs, complexes,
etc.
##STR00001##
[0029] In some embodiments the oral liquid colchicine formulation
is stable at room temperature for at least 3 months, at least 6
months, at least 18 months, or at least 24 months. In some
embodiments the oral liquid colchicine formulation is stable at
accelerated temperatures for at least 1 month, at least 2 months,
at least 3 months, or at least 6 months. In some embodiments the
oral liquid colchicine formulation is determined to be stable when
the solution has less than 5%, less than 4%, less than 3%, less
than 2%, less than 1% or less than 0.5% of any one degradant. In
other embodiments the oral liquid colchicine formulation is
determined to be stable when the solution has less than 5%, less
than 4%, less than 3%, less than 2%, less than 1% or less than 0.5%
of total degradants. In some embodiments, degradants include .beta.
lumicolchicine, .gamma.-lumicolchicine, colchiceine, and any other
individual unknown impurities.
Gout
[0030] Gout (or gouty arthritis) is a disease caused by a build-up
of uric acid due to an overproduction of uric acid or a reduced
ability of the kidney to get rid of uric acid. It is more common in
males, postmenopausal women, and people with high blood pressure.
Heavy alcohol use, diabetes, obesity, sickle cell anemia, and
kidney disease also increase the risk. The condition may also
develop in people who take drugs that interfere with uric acid
excretion.
[0031] In gout, monosodium urate or uric acid crystals are
deposited on the articular cartilage of joints, tendons and
surrounding tissues due to elevated concentrations of uric acid in
the blood stream. This provokes an inflammatory reaction of these
tissues. Gout is characterized by excruciating, sudden, unexpected,
burning pain, as well as swelling, redness, warmness, and stiffness
in the affected joint. Low-grade fever may also be present. The
patient usually suffers from two sources of pain. The crystals
inside the joint cause intense pain whenever the affected area is
moved. The inflammation of the tissues around the joint also causes
the skin to be swollen, tender and sore if it is even slightly
touched. Acute gouty arthritis (alternatively referred to as a gout
flare or a gout attack) is a sudden attack of pain in affected
joints, especially in the feet and legs. Chronic gout involves
repeated attacks of joint pain.
[0032] In acute gouty arthritis, symptoms develop suddenly and
usually involve only one or a few joints. The big toe, knee, or
ankle joints are most often affected. The pain frequently starts
during the night and is often described as throbbing, crushing, or
excruciating. The joint appears infected with signs of warmth,
redness, and tenderness. The attacks of painful joints may go away
in several days, but may return from time to time. Subsequent
attacks usually last longer. Some people may progress to chronic
gout (chronic gouty arthritis), while others may have no further
attacks.
[0033] If several attacks of gout occur each year, it can lead to
joint deformity and limited motion in joints. Uric acid deposits,
called tophi, develop in cartilage tissue, tendons, and soft
tissues. These tophi usually develop only after a patient has
suffered from the disease for many years. Deposits also can occur
in the kidneys, leading to chronic kidney failure.
[0034] Colchicine can be used for treating adults with acute gouty
arthritis and pain in attacks of acute gouty arthritis, and also
can be used beneficially for treating adults with chronic gout for
prophylaxis of acute gout flares. Although its exact mode of action
in the relief of gout is not completely understood, colchicine is
known to decrease the inflammatory response to urate crystal
deposition by inhibiting migration of leukocytes, to interfere with
urate deposition by decreasing lactic acid production by
leukocytes, to interfere with kinin formation and to diminish
phagocytosis and the subsequent anti-inflammatory response. The
anti-inflammatory effect of colchicine is relatively selective for
acute gouty arthritis. However, other types of arthritis
occasionally respond. It is neither an analgesic nor a uricosuric
and will not prevent progression to chronic gouty arthritis. It
does have a prophylactic, suppressive effect that helps to reduce
the incidence of acute attacks and to relieve the residual pain and
mild discomfort that patients with gout occasionally experienced.
In some instances, non-steroidal anti-inflammatory drugs (NSAIDs)
may also be prescribed to relieve pain and inflammation in acute
gouty arthritis attacks. Strong painkillers, such as codeine, or
corticosteroids may also be prescribed to relieve the pain.
[0035] Colchicine is rapidly absorbed from the gastrointestinal
tract. Peak concentrations occur in 0.5 to 2 hours. The drug and
its metabolites are distributed in leukocytes, kidneys, liver,
spleen and the intestinal tract. Colchicine is metabolized in the
liver and excreted primarily in the feces with 10 to 20% eliminated
unchanged in the urine. In some embodiments, oral liquid
formulations disclosed herein are used to treat gout.
Familial Mediterranean Fever (FMF)
[0036] Familial Mediterranean Fever (FMF) is a recessively
inherited disorder characterized by dramatic episodes of fever,
serosal inflammation and abdominal pain. This inflammatory disorder
is episodic, with self-limited bouts of fever accompanied by
unexplained arthritis, sterile peritonitis, pleurisy and/or skin
rash. Patients often develop progressive systemic amyloidosis from
the deposition of the acute phase reactant serum amyloid A (SAA).
In some patients, progressive systemic amyloidosis can lead to
kidney failure and death. The factors which incite an episode are
unclear. In some embodiments, colchicine can be prescribed as an
anti-inflammatory therapy.
[0037] FMF is observed primarily in individuals of non-Ashkenazi
Jewish, Armenian, Arab and Turkish background. Although rare in the
United States, incidence of FMF in Middle Eastern populations can
be as high as 1:7 in Armenian populations and 1:5 in non-Ashkenazi
Jewish populations.
[0038] FMF attacks are characterized by a massive influx of
polymorphonuclear leukocytes (PMNs) into the affected anatomic
compartment. At the biochemical level, patients have been reported
to have abnormal levels of C5a inhibitor (Matzner and Brzezinski,
"C5a-inhibitor deficiency in peritoneal fluids from patients with
familial Mediterranean fever," N. Engl. J. Med., 311:287-290
(1984)), neutrophil-stimulatory dihydroxy fatty acids (Aisen et al,
"Circulating hydroxy fatty acids in familial Mediterranean fever,"
Proc. Natl. Acad. Sci. USA, 2:1232-1236 (1985)), and dopamine
O-hydroxylase (Barakat et al, "Plasma dopamine beta-hyroxylase:
rapid diagnostic test for recurrent hereditary polyserositis,"
Lancet, 2:1280-1283 (1988)). Although linkage studies have placed
the gene causing FMF (designated MEFV) on chromosome 16p (Pras et
al., "Mapping of a gene causing familial Mediterranean fever to the
short arm of chromosome 16," N. Engl. J. Med., 326:1509-1513
(1992); Shohat et al., "The gene for familial Mediterranean fever
in both Armenians and non-Ashkenazi Jews is linked to the
.alpha.-globin complex on 16p: evidence for locus homogeneity," Am.
J. Hum. Genet., 51:1349-1354 (1992); Pras et al, "The gene causing
familial Mediterranean fever maps to the short arm of chromosome 16
in Druze and Moslem Arab families," Hum. Genet., 94:576-577 (1994);
French FMF Consortium, "Localization of the familial Mediterranean
fever gene (FMF) to a 250 kb-interval in non-Ashkenazi Jewish
founder haplotypes," Am. J. Hum. Genet., 59:603-612 (1996)), the
genetic basis of FMF has not previously been identified. In some
embodiments, oral liquid formulations disclosed herein are used to
treat FMF.
Behcet's Disease
[0039] Behcet's disease is a chronic multisystem disease
characterized by oral and genital aphthae, arthritis, cutaneous
lesions, and ocular, gastrointestinal, and neurologic
manifestations. It was first described by the Turkish dermatologist
Hulusi Behcet in 1937 as "recurrent oral aphthous ulcers, genital
ulcers, and `hypopyon-uveitis.`" The diagnosis of Behcet's disease
is based on clinical criteria as established by O'Duffy and
Goldstein and the International Study Group. Complex aphthosis is
the presence of almost constant, multiple oral or oral and genital
aphthae in the absence of systemic manifestations. These patients
must be distinguished from those with Behcet's disease. Colchicine
has been used as a treatment for Behcet's disease through its
ability to inhibit of neutrophil functions (Hirohata et al.,
Behcet's disease. Arthritis Res Ther 2003 5:139 DOI:
10.1186/ar757). In some embodiments, oral liquid formulations
disclosed herein are used to treat Behcet's Disease.
[0040] The prevalence of Behcet's disease is higher in the Middle
East and Japan where it is approximately 1 in 1000. The disease is
far less common in northern Europe, the United States, and the
United Kingdom. The mean age of onset ranges from the mid to late
20s to the fourth decade, according to several series, with a
slightly higher male to female ratio. It is relatively rare in
children and the elderly. Behcet's disease is also uncommon among
black Africans who, when they are affected, tend to have more
mucocutaneous features. Although a definitive pattern of
inheritance has not been elucidated, familial cases have been
reported. Patients with complex aphthosis are probably a subset of
patients with recurrent aphthous stomatitis, which is defined as
the recurrence of 1 or more painful oral ulcers at intervals
ranging from days to months. The prevalence of recurrent aphthosis
ranges from 5% to 66%. Onset may occur in childhood or adolescence
and some patients experience a decrease in frequency with advancing
age. (source: J. V Ghate and J. L. Jorizzo, "Behcet's disease and
complex aphthosis", Journal of the American Academy of Dermatology,
1999, 40(1), 1-18.)
Cardiovascular Disease (Atrial Fibrillation, Pericarditis)
[0041] Cardiovascular disease (CVD) involves the heart of blood
vessels. CVD includes, but is not limited to coronary artery
diseases (CAD), stroke, hypertensive heart disease, rheumatic heart
disease, cardiomyopathy, atrial fibrillation, congenital heart
disease, endocarditis, pericarditis, aortic aneurysms, peripheral
artery disease, and venous thrombosis.
[0042] One very typical and dangerous arrhythmia is atrial
fibrillation (AFIB). AFIB is the most common cardiac arrhythmia
resulting in hospitalization in the United States. AFIB is
identified by irregular heart rhythms and is clinically defined as
uncoordinated contractions of the atria. Patients often experience
palpitations and have an increased risk of stroke. Some patients
may be asymptomatic. Approximately one-third of all strokes are due
to AFIB. Furthermore, the presence of AFIB makes strokes 5-times
more likely and 2-times more debilitating.
[0043] The role of colchicine in inflammation, microtubule
disruption, adhesion of neutrophils, and other qualities, makes it
a promising treatment for some cardiovascular diseases (Deftereos
et al., Colchicine and the Heart: pushing the envelope. J Am Coll
Cardiol. 2013; 62(20):1817-1825. doi:10.1016/j.jacc.2013.08.726;
Tong et al., Colchicine in cardiovascular disease: an ancient drug
with modern tricks. 2016 Heart doi:10.1136/heartjn1-2015-309211).
In some embodiments, oral liquid formulations disclosed herein are
used to treat cardiovascular diseases (e.g., atrial fibrillation
and pericarditis).
Amyloidosis
[0044] Amyloidosis is a rare and potentially fatal disease that can
be either localized or systemic. There are four major types of
amyloidosis. The four major types include immunoglobulin (primary)
amyloidosis, reactive (secondary) amyloidosis, beta-2 microglobulin
amyloidosis and hereditary amyloidosis. Each different type of
amyloidosis presents a different prognosis and stems from different
underlining conditions. The pathologic features of amyloid deposits
include beta-pleated sheet structures that are composed of amyloid
fibrils with diameters between 8 to 10 nm. Beta-pleated sheets can
be viewed under polarized light after being stained using Congo Red
stain, these stained fibrils display an apple green
birefringence.
[0045] Secondary amyloidosis is associated with chronic
inflammatory diseases such as FMF. The precursor protein
responsible for constructing the amyloid fibrils associated with
secondary amyloidosis is serum amyloid A, an acute-phase reactant.
Typical sites of amyloid accumulation include the spleen, liver,
lymph nodes, adrenal glands, and the kidneys. Symptoms that are
nonspecific include complaints of weakness and fatigue. Specific
complaints are directly associated to organ involvement, these
symptoms commonly include edema and pain. In some embodiments, oral
liquid formulations disclosed herein are used to treat
amyloidosis.
Calcium Pyrophosphate Deposition Disease (Pseudogout)
[0046] Calcium pyrophosphate deposition disease (CPDD), also known
as pseudogout, chondrocalcinosis, and pyrophosphate arthropathy, is
a rheumatologic disorder with varied symptoms and signs arising
from the accumulation of crystals of calcium pyrophosphate
dihydrate in the connective tissues.
[0047] Pseudogout refers to the acute symptoms of joint
inflammation or synovitis: red, tender, and swollen joints that may
resemble gouty arthritis. The disorder is more common in older
adults. It may be asymptomatic, or it can be associated with
osteoarthritis, or it can present as an acute or chronic
inflammatory arthritis that causes pain in one or more joints. The
white blood cell count is often raised.
[0048] The arthritis is usually polyarticular (inflammation of
several joints in the body), although it may begin as monoarticular
(one joint). CPPD crystals tend to form within articular tissues.
Knees are the most commonly affected joints, along with wrists and
hips. In rare cases, pseudogout may affect the spinal canal and
cause damage to the spinal cord. In some embodiments, oral liquid
formulations disclosed herein are used to treat pseudogout.
Cirrhosis of the Liver
[0049] Cirrhosis, a condition in which the liver does not function
properly due to long-term damage, typically comes on slowly over
months or years. Early on, there are often no symptoms. As the
disease worsens, a subject may become tired, weak, itchy, have
swelling in the lower legs, develop yellow skin, bruise easily,
have fluid build-up in the abdomen, or develop spider-like blood
vessels on the skin. The fluid build-up in the abdomen may become
spontaneously infected. Other complications include hepatic
encephalopathy, bleeding from dilated veins in the esophagus or
dilated stomach veins, and liver cancer. Hepatic encephalopathy
results in confusion and possibly unconsciousness. Colchicine has
been shown to have anti-fibrotic effects in relation to hepatic
diseases (Leung et al., Colchicine--Update on mechanisms of action
and therapeutic uses. 2015. Seminar in Arthritis and Rheumatism. 45
(3), 257-67).
[0050] Cirrhosis is most commonly caused by alcohol, hepatitis B,
hepatitis C, and non-alcoholic fatty liver disease. Typically, more
than two or three drinks per day over a number of years is required
for alcoholic cirrhosis to occur. Non-alcoholic fatty liver disease
is due to a number of reasons, including being overweight,
diabetes, high blood fats, and high blood pressure. A number of
less common causes include autoimmune hepatitis, primary biliary
cirrhosis, hemochromatosis, certain medications, and gallstones.
Cirrhosis is characterized by the replacement of normal liver
tissue by scar tissue. These changes lead to loss of liver
function. Diagnosis is based on blood testing, medical imaging, and
liver biopsy. In some embodiments, oral liquid formulations
disclosed herein are used to treat hepatic diseases (e.g.,
cirrhosis of the liver).
Sarcoid Arthritis
[0051] Sarcoidosis, a disease involving abnormal collections of
inflammatory cells, can be involved with the joints, bones and
muscles. This causes a wide variety of musculoskeletal complaints
that act through different mechanisms. Approximately 5-15% of cases
affect the bones, joints, or muscles.
[0052] Sarcoid arthritis has two classifications: acute or chronic.
Sarcoidosis patients with acute arthritis often also accompanies
bilateral Hilar lymphadenopathy and Erythema nodosum. Usually true
arthritis is not present, but instead periarthritis presents itself
as a swelling in the soft tissue around the joints that can be seen
by ultrasonographic methods. These joint symptoms tend to precede
or occur at the same time as erythema nodosum develops. Enthesitis
also occurs in about one-third of patients with acute sarcoid
arthritis, mainly affecting the Achilles tendon and heels. Soft
tissue swelling at the ankles can be prominent, and biopsy of this
soft tissue reveals no granulomas, but does show panniculitis that
is similar to erythema nodosum.
[0053] Chronic sarcoid arthritis usually occurs in the setting of
more diffuse organ involvement. The ankles, knees, wrists, elbows,
and hands may all be affected in the chronic form and often in a
polyarticular pattern. Dactylitis similar to that seen in Psoriatic
arthritis, that is associated with pain, swelling, overlying skin
erythema, and underlying bony changes may also occur. In some
embodiments, oral liquid formulations disclosed herein are used to
treat sarcoid arthritis.
Disk Diseases & Related Spinal Disorders
[0054] Disk diseases & related spinal disorders are a group of
disorders that are quite painful. It is believed that colchicine
acts directly on diskal inflammation to reduce inflammation in the
area surrounding the spinal nerve roots. Colchicine has also been
shown to cause an increase of endorphin-producing neurons in the
spinal cord and to prevent deposition of amyloid in damaged disk.
In some embodiments the subject has diskal back pain and or
sciatica. In some embodiments, oral liquid formulations disclosed
herein are used to treat disk diseases and related spinal
disorders.
Formulations
[0055] The liquid formulations described herein may include
additional ingredients. For instance these additional components
may include, but are not limited to, buffering agents,
preservatives, sweeteners, flavoring agents, glycols such as
propylene glycol and glycerin, as examples, and coloring agents.
Additional excipients such as tonicity agents and chelating agents
are within the scope of the embodiments.
[0056] Buffering agents maintain the pH when colchicine is
compounded into a liquid form. Non-limiting examples of buffering
agents include, but are not limited to, sodium bicarbonate,
potassium bicarbonate, magnesium hydroxide, magnesium lactate,
magnesium gluconate, aluminum hydroxide, aluminum hydroxide/sodium
bicarbonate precipitate, a mixture of an amino acid and a buffer, a
mixture of aluminum glycinate and a buffer, a mixture of acid salt
and an amino acid and a buffer, and a mixture of an alkali salt of
an amino acid and a buffer. Additional buffering agents include
citric acid, sodium citrate, sodium tartarate, sodium acetate,
sodium carbonate, sodium polyphosphate, potassium polyphosphate,
sodium pyrophosphate, potassium pyrophosphate, disodium hydrogen
phosphate, dibasic sodium phosphate, trisodium phosphate,
tripotassium phosphate, sodium acetate, potassium metaphosphate,
magnesium oxide, magnesium carbonate, magnesium silicate, calcium
acetate, calcium glycerophosphate, calcium chloride, calcium
hydroxide, calcium lactate, calcium carbonate, calcium bicarbonate,
and other calcium salts. Some buffering agents also impart
effervescent qualities when a powder is compounded into a liquid.
In some embodiments, the colchicine described herein, when
compounded into a liquid form, comprises a buffering agent.
[0057] Preservatives include anti-microbials, anti-oxidants, and
agents that enhance sterility. Exemplary preservatives include
ascorbic acid, ascorbyl palmitate, benzyl alcohol, BHA, BHT, citric
acid, erythorbic acid, fumaric acid, malic acid, propyl gallate,
sodium ascorbate, sodium benzoate, sodium bisulfate, sodium
metabisulfite, sodium sulfite, parabens (methyl-, ethyl-, butyl-),
benzoic acid, potassium sorbate, and vanillin. In some embodiments,
the colchicine described herein, when compounded into a liquid
form, comprises a preservative.
[0058] Sweeteners or sweetening agents include any compounds that
provide a sweet taste to make the product more palatable. This
includes natural and synthetic sugars, natural and artificial
sweeteners (e.g., sucralose), natural extracts and any material
that initiates a sweet sensation in a subject. In some embodiments,
the colchicine described herein, when compounded into a liquid
form, comprises a sweetener. In other embodiments, sweeteners in
liquid form are used to solvate or dissolve the colchicine
described herein.
[0059] Sugars illustratively include glucose, fructose, sucrose,
xylitol, tagatose, maltitol, isomaltulose, lactitol, sorbitol,
mannitol, erythritol, trehalose, maltodextrin, polydextrose, and
the like. Other sweeteners include glycerin, inulin, maltol,
acesulfame and salts thereof, e.g., acesulfame potassium, alitame,
aspartame, neotame, sodium cyclamate, saccharin and salts thereof,
e.g., saccharin sodium or saccharin calcium, neohesperidin
dihydrochalcone, stevioside, thaumatin, and the like. Sweeteners
can be used in the form of crude or refined products such as
hydrogenated starch hydrosylates, maltitol syrup, high fructose
corn syrup, and as branded proprietary blend products. Sweeteners
can be used singly or combinations of two or more. Suitable
concentrations of different sweeteners can be selected based on
published information, manufacturers' data sheets, and by routine
testing. In certain instances, an above-described flavored solution
component is used to solvate or dissolves colchicine described
herein.
[0060] In another embodiment, the liquid form comprises a flavoring
agent or flavorant to enhance the taste or aroma of the solution
component used to solvate or dissolve the colchicine described
herein. Suitable natural or synthetic flavoring agents can be
selected from standard reference books, such as Remington: The
Science and Practice of Pharmacy (2000) and Fenaroli's Handbook of
Flavor Ingredients (1994). Non-limiting examples of suitable
natural flavors, some of which can be readily simulated with
synthetic agents or combinations thereof, include almond, anise,
apple, apricot, banana, blackberry, blackcurrant, blueberry,
caramel, cherry, chocolate, cinnamon, cranberry, grape, lemon,
lime, orange, peppermint, pineapple, raspberry, spearmint,
strawberry, vanilla, etc. Also useful, particularly where the
composition is intended primarily for pediatric use is tutti-frutti
or bubble gum flavor, a compounded flavoring agent based on fruit
flavors. Presently, preferred flavoring agents include bubble gum,
strawberry, cherry, grape, orange, peppermint, and vanilla. In some
embodiments, the resultant liquid form from the colchicine
described herein comprises a Flavor Cherry 825.662 flavoring agent.
Flavoring agents may be used singly or in combinations of two or
more.
[0061] In further embodiments, the resultant liquid form from the
colchicine described herein comprises a coloring agent for identity
and/or aesthetic purposes. Suitable coloring agents approved by the
U.S. Food and Drug Administration (FDA) include FD&C Red No. 3,
FD&C Red No. 20, FD&C Red No. 40, FD&C Yellow No. 6,
FD&C Blue No. 2, D&C Green No. 5, D&C Orange No. 5,
D&C Yellow No. 10, caramel, ferric oxide and mixtures
thereof.
[0062] In further embodiments, the resultant liquid form from the
colchicine described herein comprises a thickening agent Thickening
agents include, but are not limited to xanthan gum.
[0063] In some embodiments other flavoring agents, buffering
systems, and preservatives may be used. The solution is formulated
to inhibit growth of bacteria, mold, and yeast for storage at room
temperature and ambient conditions.
[0064] In some embodiments, the liquid formulation is by % w/v can
be found in Table 1. In other embodiments, the formulation includes
0.2-0.4% w/v of benzyl alcohol, 0.1-0.3% w/v of anhydrous citric
acids, 0.005-0.025% w/v of colchicine, 0.005-0.02% w/v of FD&C
Red No. 40, 0.8-1.6% w/v or dibasic sodium phosphate, heptahydrate,
0.75-0.15% w/v of flavor cherry 825.662, 2-8% w/v of propylene
glycol, 2-10% glycerin, 0.1-0.2% w/v of sucralose, 0.1-0.2% w/v of
xanthan gum, and water. In other embodiments, the formulation
includes 0.28-3.2 or 0.3% w/v of benzyl alcohol, 0.2% w/v of
anhydrous citric acids, 0.012% w/v of colchicine, 0.01% w/v of
FD&C Red No. 40, 1.2% w/v or dibasic sodium phosphate,
heptahydrate, 0.125% w/v of flavor cherry 825.662, 5% w/v of
propylene glycol, 0.15% w/v of sucralose, 0.15% w/v of xanthan gum,
and water.
TABLE-US-00002 TABLE 1 Exemplary formulation Ingredient % w/v
Benzyl Alcohol 0.3 Citric Acid, Anhydrous 0.2 Colchicine * 0.012
FD&C Red No. 40 0.01 Dibasic Sodium Phosphate, Heptahydrate 1.2
Flavor Cherry 825.662 0.125 Propylene Glycol 5 Glycerin 5 Sucralose
0.15 Xanthan Gum 0.15 Water Q.S. * Calculated on the anhydrous,
solvent free basis
Storage
[0065] The colchicine described herein is stable in various storage
conditions including refrigerated, ambient, and accelerated
conditions. Stable as used herein refers to the ability of an
active agent to maintain activity under standard stability
conditions. Standard stability conditions include relative humidity
conditions along with the temperatures, 25 degrees C. 60% RH(RT),
30 C 65% RH (ICH), and 40 C 75% RH (accelerated), for example.
[0066] At refrigerated and ambient conditions, the liquid
colchicine composition described herein in stable for at least 1
month, at least 2 months, at least 3 months, at least 6 months, at
least 9 months, at least 12 months, at least 15 months, at least 18
months, and at least 24 months. At accelerated conditions, the
colchicine solution described herein is stable for at least 1
month, at least 2 months, at least 3 months and at least 6 months.
Accelerated conditions include temperatures that are above ambient
levels. In some instances, an accelerated condition is at about
30.degree. C., about 35.degree. C., about 40.degree. C., about
45.degree. C., about 50.degree. C., about 55.degree. C., or about
60.degree. C. Ambient conditions include temperature that is at
ambient levels. In some instances, an ambient condition is at about
20.degree. C., about 21.degree. C., about 22.degree. C., about
23.degree. C., about 24.degree. C., about 25.degree. C., about
26.degree. C., about 27.degree. C., about 28.degree. C., about
29.degree. C., and about 30.degree. C. Refrigerated conditions
include temperature in typical refrigeration units (e.g.
5.+-.3.degree. C.). In some instances, a refrigerated condition is
about 2.degree. C., about 3.degree. C., about 4.degree. C., about
5.degree. C., about 6.degree. C., about 7.degree. C., or about
8.degree. C.
[0067] Liquid vehicles suitable for the colchicine described herein
are selected for a particular oral liquid composition (e.g.,
solution, suspension, etc.) as well as other properties such as
clarity, viscosity, compatibility with excipients, chemical
inertness, palatability, odor, and color. Exemplary liquid vehicles
include water, ethyl alcohol, glycerin, propylene glycol, syrup
(e.g., sugar or other sweetener based, e.g., Ora-Sweet.RTM. SF
sugar-free flavored syrup), juices (e.g., apple, orange, cranberry,
cherry, tomato and the like), other beverages (e.g., tea, coffee,
soft drinks, milk and the like), oils (e.g., olive, soybean, corn,
mineral, castor and the like), and combinations or mixtures
thereof. Certain liquid vehicles, e.g., oil and water, can be
combined together to form emulsions. In some embodiments, water is
used as a vehicle for a colchicine oral liquid. In other
embodiments, propylene glycol is used as a vehicle for a colchicine
oral liquid. For the liquid colchicine described herein, the
solution component is used as the vehicle for a colchicine oral
liquid.
[0068] The viscosity of the solution is an important component. In
some embodiments the solution has a viscosity in the range of
40-800 cps. In other embodiments the solution has a viscosity of
80-250 cps.
[0069] The colchicine oral liquid compositions may be used for the
treatment of diseases and conditions described herein. In addition,
a method for treating any of the diseases or conditions described
herein in a subject in need of such treatment involves
administration of colchicine oral liquid compositions in
therapeutically effective amounts to the subject. In some
embodiments, the amount of a given colchicine oral liquid
composition that corresponds to such an amount varies depending on
factors such as the particular colchicine salt or form, disease
condition and its severity, the identity (age, weight, sex) of the
subject or patient in need of treatment, but can nevertheless be
determined according to the particular circumstances surrounding
the case, including, e.g., the specific agent being administered,
the liquid composition type, the condition being treated, and the
subject or patient being treated.
[0070] In further embodiments, the daily dosages appropriate for
the colchicine oral liquid compositions described herein are from
about 0.2 mg-1.5 mg/dose/day or in other embodiments 0.5 mg-1.5
mg/dose/day. In one embodiment, the daily dosage appropriate for
the colchicine liquid compositions is about 0.6-1.2
mg/dose/day.
[0071] The treatment of certain diseases or conditions (e.g., gout,
FMF, cardiac disease etc.) in a patient or subject with a
colchicine oral liquid composition described herein encompass
additional therapies and treatment agents in some embodiments. Such
additional therapies and treatment regimens include another
therapy, e.g., antibiotics, for the treatment of the particular
disease in some embodiments.
[0072] Unless defined otherwise, all technical and scientific terms
used herein have the same meanings as commonly understood by one of
ordinary skill in the art. Although any methods and materials
similar or equivalent to those described herein can be used in the
practice of testing of embodiments described herein, certain
preferred methods, devices, and materials are now described.
[0073] As used herein and in the appended claims, the singular
forms "a", "an", and "the" include plural reference unless the
context clearly dictates otherwise. Thus, for example, reference to
"an excipient" is a reference to one or more excipients and
equivalents thereof known to those skilled in the art, and so
forth.
[0074] The term "about" is used to indicate that a value includes
the standard level of error for the device or method being employed
to determine the value. The use of the term "or" in the claims is
used to mean "and/or" unless explicitly indicated to refer to
alternatives only or the alternatives are mutually exclusive,
although the disclosure supports a definition that refers to only
alternatives and to "and/or". The terms "comprise", "have", and
"include" are open-ended linking verbs. Any forms or tenses of one
or more of these verbs "comprises," "comprising," "has," "having,"
"includes," and "including" are also open-ended. For example, any
method that "comprises," "has" or "includes" one or more steps is
not limited to possessing only those one or more steps and also
covers other unlisted steps.
[0075] "Optional" or "optionally" may be taken to mean that the
subsequently described structure, event or circumstance may or may
not occur, and that the description includes instances where the
events occurs and instances where it does not.
[0076] As used herein, the term "therapeutic" means an agent
utilized to treat, combat, ameliorate, prevent or improve an
unwanted condition or disease of a patient. In some embodiments, a
therapeutic agent such as oral colchicine is directed to the
treatment and/or the amelioration of, reversal of, or stabilization
of the symptoms of gout, familial Mediterranean fever (FMF),
pericarditis, Behcet's disease, atrial fibrillation, amyloidosis,
calcium pyrophosphate deposition disease (pseudogout), cirrhosis of
the liver, sarcoid arthritis, and inflammatory diseases described
herein.
[0077] "Administering" when used in conjunction with a therapeutic
means to administer a therapeutic systemically or locally, as
directly into or onto a target tissue, or to administer a
therapeutic to a patient whereby the therapeutic positively impacts
the tissue to which it is targeted. Thus, as used herein, the term
"administering", when used in conjunction with an oral colchicine
composition, can include, but is not limited to, providing an oral
colchicine composition into or onto the target tissue; providing an
oral colchicine composition systemically to a patient by, e.g.,
oral administration whereby the therapeutic reaches the target
tissue or cells. "Administering" a composition may be accomplished
by injection, topical administration, and oral administration or by
other methods alone or in combination with other known
techniques.
[0078] As used herein, the terms "patient," "subject" and
"individual" are intended to include living organisms in which
certain conditions as described herein can occur. Examples include
humans, monkeys, cows, sheep, goats, dogs, cats, mice, rats, and
transgenic species thereof. In a preferred embodiment, the patient
is a primate. In certain embodiments, the primate or subject is a
human. In certain instances, the human is an adult. In certain
instances, the human is child. In certain instances, the human is
elderly. In other instances, the human is 65 years of age or older.
Other examples of subjects include experimental animals such as
mice, rats, dogs, cats, goats, sheep, pigs, and cows. By
"pharmaceutically acceptable", it is meant the carrier, diluent or
excipient must be compatible with the other ingredients of the
formulation and not deleterious to the recipient thereof.
[0079] The term "pharmaceutical composition" shall mean a
composition comprising at least one active ingredient, whereby the
composition is amenable to investigation for a specified,
efficacious outcome in a mammal (for example, without limitation, a
human). Those of ordinary skill in the art will understand and
appreciate the techniques appropriate for determining whether an
active ingredient has a desired efficacious outcome based upon the
needs of the artisan.
[0080] A "therapeutically effective amount" or "effective amount"
as used herein refers to the amount of active compound or
pharmaceutical agent that elicits a biological or medicinal
response in a tissue, system, animal, individual or human that is
being sought by a researcher, veterinarian, medical doctor or other
clinician, which includes one or more of the following: (1)
preventing the disease; for example, preventing a disease,
condition or disorder in an individual that may be predisposed to
the disease, condition or disorder but does not yet experience or
display the pathology or symptomatology of the disease, (2)
inhibiting the disease; for example, inhibiting a disease,
condition or disorder in an individual that is experiencing or
displaying the pathology or symptomatology of the disease,
condition or disorder (i.e., arresting further development of the
pathology and/or symptomatology), and (3) ameliorating the disease;
for example, ameliorating a disease, condition or disorder in an
individual that is experiencing or displaying the pathology or
symptomatology of the disease, condition or disorder (i.e.,
reversing the pathology and/or symptomatology).
[0081] The terms "treat," "treated," "treatment," or "treating" as
used herein refers to both therapeutic treatment in some
embodiments and prophylactic or preventative measures in other
embodiments, wherein the object is to prevent or slow (lessen) an
undesired physiological condition, disorder or disease, or to
obtain beneficial or desired clinical results. For the purposes
described herein, beneficial or desired clinical results include,
but are not limited to, alleviation of symptoms; diminishment of
the extent of the condition, disorder or disease; stabilization
(i.e., not worsening) of the state of the condition, disorder or
disease; delay in onset or slowing of the progression of the
condition, disorder or disease; amelioration of the condition,
disorder or disease state; and remission (whether partial or
total), whether detectable or undetectable, or enhancement or
improvement of the condition, disorder or disease. Treatment
includes eliciting a clinically significant response without
excessive levels of side effects. Treatment also includes
prolonging survival as compared to expected survival if not
receiving treatment. A prophylactic benefit of treatment includes
prevention of a condition, retarding the progress of a condition,
stabilization of a condition, or decreasing the likelihood of
occurrence of a condition. As used herein, "treat," "treated,"
"treatment," or "treating" includes prophylaxis in some
embodiments.
[0082] Embodiments have been described where the techniques are
implemented in circuitry and/or computer-executable instructions.
It should be appreciated that some embodiments may be in the form
of a method, of which at least one example has been provided. The
acts performed as part of the method may be ordered in any suitable
way. Accordingly, embodiments may be constructed in which acts are
performed in an order different than illustrated, which may include
performing some acts simultaneously, even though shown as
sequential acts in illustrative embodiments.
[0083] Various aspects of the embodiments described above may be
used alone, in combination, or in a variety of arrangements not
specifically discussed in the embodiments described in the
foregoing and is therefore not limited in its application to the
details and arrangement of components set forth in the foregoing
description or illustrated in the drawings. For example, aspects
described in one embodiment may be combined in any manner with
aspects described in other embodiments.
[0084] The present invention is further illustrated by the
following Examples, which in no way should be construed as further
limiting. The entire contents of all of the references (including
literature references, issued patents, published patent
applications, and co-pending patent applications) cited throughout
this application are hereby expressly incorporated by
reference.
EXAMPLES
Example 1: Formulations
TABLE-US-00003 [0085] TABLE 2 Liquid Colchicine Formulation
Ingredient % w/v Benzyl Alcohol 0.3 Citric Acid, Anhydrous 0.2
Colchicine* 0.012 FD&C Red No. 40 0.01 Dibasic Sodium
Phosphate, Heptahydrate 1.2 Flavor Cherry 825.662 0.125 Propylene
Glycol 5 Glycerin 5 Sucralose 0.15 Xanthan Gum 0.15 Water Q.S.
*Calculated on the anhydrous, solvent free basis
[0086] In some embodiments, the oral liquid formulation includes:
the 0.3% w/v of benzyl alcohol, 0.2% w/v of anhydrous citric acids,
0.012% w/v of colchicine, 0.01% w/v of FD&C Red No. 40, 1.2%
w/v or dibasic sodium phosphate, heptahydrate, 0.125% w/v of flavor
cherry 825.662, 5% w/v of propylene glycol, 0.15% w/v of sucralose,
0.15% w/v of xanthan gum, and water as seen in Table 2.
Example 2: Stability
[0087] Colchicine has been shown to be unstable at room temperature
in solution. An article published in Drug Development and
Industrial Pharmacy, 15(11), 1905-1909 (1989) by Habib, et. al.,
investigated the stability of colchicine and showed that there is
photodegradation of colchicine in solution, especially in the
presence of glycerin, (up to a 50% loss). The additives, such as
lithium carbonate, p-aminobenzoic acid, and uric acid, used in this
study to try and prevent the degradation of the colchicine are not
acceptable excipients for an oral solution. Furthermore, these
ingredients did not prevent the degradation of the colchicine. The
stability studies described in this paper were conducted over
hours, which would not yield a pharmaceutically acceptable
product.
[0088] Colchicine solutions are used in biological studies for
cellular assays. Companies like BI (Biological Industries) sell a
colchicine solution in a cellular buffering agent. The storage
conditions are 2-8.degree. in order to prevent degradation, as
these types of solutions are not stable at room temperature.
Furthermore, the safety information/MSDS for the buffers used to
make these solutions state that it is harmful if ingested.
[0089] The product information from various suppliers of colchicine
crystalline powder for laboratory use state that colchicine should
be stored at freezing temperatures or refrigeration to maintain
stability. Cayman Chemical's product information regarding
colchicine states that they do not recommend storing an aqueous
solution made from their crystalline colchicine powder for more
than one day. Sigma product information on colchicine says to store
any made solutions between 2-8.degree. for stability purposes.
Furthermore these product information sheets are for the colchicine
crystalline powder and any solutions described are not only not
stable at room temperature for any period of time, but are not
solutions for oral consumption, as they are made with buffers and
solvents that are not for human consumption, such as DPBS, benzene,
and chloroform. These solutions are for cellular assays in a
laboratory setting.
[0090] The colchicine injections for IV administration are made in
single unit doses, and the label and package inserts for these
products specifically state that the colchicine injections are for
intravenous use only.
[0091] The colchicine compounded solutions for veterinary purposes
are for immediate use, and there is no stability data generated on
these types of products. As the prior art shows, colchicine
solutions currently manufactured are not stable at room temperature
for long term, and furthermore none of the solutions described
could be used for human oral consumption.
[0092] Surprisingly, a stable oral colchicine solution that is even
stable in the presence of glycerin has been developed according to
the invention. The colchicine solution is formulated and assayed
for colchicine levels. Stability of the claimed formulation was
tested at room temperature, 30.degree. C., and 40.degree. C. and
assayed for the colchicine. Results can be found in Table 3 below.
Table 4 shows further stability data up through 6 months. The fact
that the oral solution was so stable after 6 months at an
accelerated temperature (40 degrees Celsius, which corresponds to
24 months at room temperature) was quite surprising. Solutions of
colchicine for oral administration to humans have not been
developed because they were believed to be too unstable. The data
presented herein show for the first time that an oral colchicine
solution can remain stable with minimal degradants and thus have a
long shelf life consistent with commercial formulations.
TABLE-US-00004 TABLE 3 Stability Results of Colchicine Solution (%
Label Claim) Sample Result API (colchicine drug substance)* 91.8%
3.5 months @ 5.degree. 92.3% 3 months @ 25.degree. .+-. 2.degree.
C. 60% .+-. 5% RH(RT) 92.6% 3 months @ 30.degree. .+-. 2.degree. C.
65% .+-. 5% RH(ICH) 91.3% 3 months @ 40.degree. .+-. 2.degree. C.
75% .+-. 5% RH(ACC) 91.1% *Calculated on the anhydrous,
solvent-free basis
TABLE-US-00005 TABLE 4 Stability Report for Colchicine Solution,
0.12 mg/mL Test Conditions Initial 1 Month 2 Months 3 Months 6
Months 25.degree. C./60% RH 98.1% -- 96.7% 97.3% 96.7% 30.degree.
C./65% RH 98.1% -- 96.8% 96.5% 96.6% 40.degree. C./75% RH 98.1%
96.6% 97.0% 96.7% 95.1%
[0093] These results show that there is no loss of colchicine from
the initial raw API used and under all conditions after 6 months.
These results show that the colchicine solution of the invention is
stable, even at high temperatures. The colchicine does not degrade
over time and under accelerated conditions. The results were
calculated on an as is basis. The colchicine solution in Table 4
was packaged in a 190 mL HDPE bottle and assayed by HPLC cUSP.
[0094] These results show the percent of label claim of colchicine
in the solution of the invention. The label claim of the liquid
solution is 0.12 mg/ml and the assay results of the active
ingredient are reported as percent of label claim. For instance,
the label claim of an exemplary colchicine solution of the
invention is 0.12 mg/ml. The FDA requirement for stability results
for drugs is 90-110% label claim. These results show that
colchicine was not degraded over time under accelerated conditions
as well as at room temperature, which is surprising based on the
prior art.
[0095] The pH of the liquid solution was evaluated (Table 5). The
results show that the solution maintained a stable pH at
accelerated conditions as well at room temperature.
TABLE-US-00006 TABLE 5 Evaluation of pH of Colchicine Oral Solution
(Initial pH = 6.6) Condition 1 month 2 months 3 months RT 6.48 6.61
6.59 30.degree. 6.59 6.57 6.60 40.degree. 6.49 6.59 6.57
[0096] A desirable range of pH includes 6.2-7.2. In some
embodiments the pH range is 6.3-6.7.
[0097] Tables 6 and 7 below show data from USP antimicrobial
preservatives effectiveness tests. The product tested in both cases
met the requirements of the Current USP for Oral Products (Category
3 Products).
TABLE-US-00007 TABLE 6 Day 14 Day 28 Test Initial Log red Log red
Organism Inoculum/ml CFU/ml (% red) CFU/ml (% red) S. aureus 6.1
.times. 10.sup.5 <10 >4 <10 >4 ATCC #6538 (>99.99)
(>99.99) E.coli 5.8 .times. 10.sup.5 1.98 .times. 10.sup.4 1.47
1.46 .times. 10.sup.3 2.60 ATCC #8739 (96.59) (>99.75) Ps.
aeruginosa 8.6 .times. 10.sup.5 <10 >4 <10 >4 ATCC
#9027 (>99.99) (>99.99) C. albicans 3.5 .times. 10.sup.5 6.0
.times. 10.sup.3 1.77 <10 >4 ATCC #10231 (98.29) (>99.99)
A. brasiliensis 3.5 .times. 10.sup.5 1.1 .times. 10.sup.2 3.50
<10 >4 ATCC #16404 (99.97) (>99.99)
TABLE-US-00008 TABLE 7 Day 14 Day 28 Test Initial Log red Log red
Organism Inoculum/ml CFU/ml (% red) CFU/ml (% red) S. aureus 6.1
.times. 10.sup.5 5.4 .times. 10.sup.3 2.05 <10 >4 ATCC #6538
(99.11) (>99.99) E.coli 5.8 .times. 10.sup.5 6.5 .times.
10.sup.1 3.95 <10 >4 ATCC #8739 (99.99) (>99.99) Ps.
aeruginosa 8.6 .times. 10.sup.5 <10 >4 <10 >4 ATCC
#9027 (>99.99) (>99.99) C. albicans 3.5 .times. 10.sup.5 4.6
.times. 10.sup.2 2.88 <10 >4 ATCC #10231 (99.87) (>99.99)
A. brasiliensis 3.5 .times. 10.sup.5 1.1 .times. 10.sup.2 3.50
<10 >4 ATCC #16404 (99.97) (>99.99)
Example 3: Colchicine Assay
[0098] This example presents the results of the evaluation of the
application of the current USP Official (12/1/15-4/30/16) Monograph
for Colchicine/Colchicine Injection
[0099] (Assay) for Colchicine Oral Liquid 0.12 mg/mL. Evaluation
included demonstration of system suitability, specificity for
placebo and potentially interfering individual placebo components,
and analysis of a sample preparation. The analysis was performed
and documented within QCL project G8968.
[0100] Table 8, below, shows the sample materials that were used
for the evaluation.
TABLE-US-00009 TABLE 8 Sample Materials Material Description Lot #
Colchicine Oral Liquid, 0.12 mg/mL Formula 0090A-1 011416B
Colchicine Oral Liquid Placebo Formula 0090A-1-PI 011416A
Colchicine, USP API (Indena) 30670/H FD&C Red No. 40 47943
Flavor Cherry 825.662 FONA S1528930 Benzyl Alcohol Sigma-Aldrich
SHBD7983V
[0101] The current USP Assay method for Colchicine Injection refers
to the Assay method described in the monograph for Colchicine drug
substance. The method employs an isocratic reverse phase separation
with stationary phase L7 (C8) and UV detection at 254 nm with a 20
.mu.L injection volume. The sample was prepared at a target
concentration of 6 .mu.g/mL Colchicine and compared to an external
standard prepared at the same concentration. For the purposes of
evaluation, Colchicine API was used for preparation of the external
standard. Standard solutions were prepared in duplicate by the
dilution of 12 mg Colchicine API to 200.0 mL in 50:50
methanol:water diluent. 10.0 mL of the resulting stock standard was
further diluted to 100.0 mL in diluent to a concentration of 6
.mu.g/mL Colchicine working standard. For the preparation of the
sample solution, 8.0 mL (equivalent to 0.96 mg Colchicine) of the
oral liquid was diluted to 50.0 mL in diluent. 30.0 mL of the
resulting solution was further diluted to 100.0 mL in diluent to a
concentration of 6 .mu.g/mL Colchicine working sample solution. The
HPLC method parameters are summarized in Table 9 and example
chromatography is provided in FIG. 1.
TABLE-US-00010 TABLE 9 HPLC Parameters Parameter Mobile Phase 45:55
0.5M monobasic potassium phosphate:methanol, pH 5.5 Column Waters
SPHERISORB .RTM. 5 .mu.m C8 4.6 .times. 250 mm Flow Rate 1.1 mL/min
Column Temperature 21.degree. C. Injection Volume 20 .mu.L
Detection UV 254 nm
[0102] All specified system suitability parameters were met and
maintained throughout the analysis. A summary of system suitability
requirements and results are provided in Table 10.
TABLE-US-00011 TABLE 10 System Suitability Parameters Parameter
Requirement Result Column efficiency (Theoretical Plates) NLT 4500
7640 Retention Time-Colchicine 5.5-9.5 minutes 8.8 minutes RSD
Colchicine Area NMT 2% 0.0% Standard Check Agreement 98.0-102.0%
99.6%
[0103] Colchicine Oral Liquid, 0.12 mg/mL, Formula 0090A-1, and the
liquid placebo includes ingredients at the concentrations shown in
Table 5. Placebo and ingredients with the potential for UV
absorptivity and chromatographic interference were examined (Table
11).
TABLE-US-00012 TABLE 11 Ingredient % % w/v Potential for
Interference (Y/N) Benzyl Alcohol 0.3 Y Citric Acid, Anhydrous 0.2
N Colchicine 0.012* -- FD&C Red No. 40 0.01 Y Dibasic Sodium
Phosphate, 1.2 N Heptahydrate Flavor Cherry 825.662 0.125 Y
Glycerin USP (99.7%) 5.0 N Propylene Glycol 5.0 N Sucralose 0.15 N
Xanthan Gum 0.15 N Water Q. S. N *Calculated on the anhydrous,
solvent free basis
[0104] The placebo formulation and individual ingredients benzyl
alcohol, FD&C Red No. 40, and Flavor Cherry 825.662 were
prepared at nominal (100% of target formulation) concentrations in
diluent. An overlay of the resulting peaks including chromatograms
of diluent blank and colchicine standard are included in FIG. 2.
Injections of benzyl alcohol and FD&C Red No. 40 resulted in
single peaks that did not interfere at the retention time of
colchicine. For the injection of Flavor Cherry 825.662, a primary
peak and four minor secondary peaks were recorded. The secondary
flavor related peak, at a retention time of 8.9 minutes, elutes at
the approximate retention time of Colchicine (8.8 minutes),
contributing interference at approximately 1.5% of the area
response of a standard injection. Because the interfering peak
completely co-elutes with colchicine, the assay calculation was
performed by subtraction of the flavor related peak response
obtained from injection of the placebo solution from the colchicine
response of the sample injections.
[0105] The sample preparation was injected in duplicate, with the
calculation performed against bracketing standard injections. The
area response of the flavor related peak at retention time 8.9
minutes was subtracted from the area response of Colchicine in the
sample injections. The assay results obtained was 99.1% of the
labeled amount.
[0106] The USP Colchicine Assay method evaluated as described in
this report is suitable for the routine assay of Colchicine in
Colchicine Oral Liquid, 0.12 mg/mL. Minor interference due to the
flavor cherry ingredient was mitigated by concurrent injection of
placebo sample with area subtraction.
Example 4: Caco-2 Permeability of Colchicine
[0107] A Caco-2 study was conducted to compare the permeability of
a colchicine solution of the invention to commercially available
tablet and capsules. This study is an in vitro test that uses a gut
cell line to measure permeability of the test articles. The results
show that the colchicine oral solution has a similar permeability
as the tablets and the capsules. These results indicate that the
bioavailability of the colchicine solution in vivo will be similar
to the tablets and the capsules.
[0108] The purpose of this study was to determine the permeability
of Colchicine from different formulations for the purpose of BCS
classification. This study was performed under non-GLP conditions.
All work was performed with appropriate local health regulations
and ethical approval. Colchicine monoisotopic mass is 399.17, and
parent MW (free base) is 399.44.
Procedure:
[0109] Colchicine-Liquid--1 bottle
[0110] Colchicine Liquid (1 mL equivalent to 300 .mu.M Colchicine)
was tested. The procedure included shaking the colchicine liquid
bottle for a few seconds before using. The solution was then
diluted to the final assay concentration.
Colchicine Tablets, USP+Sterile Water for Injection
[0111] Colchicine tablets, USP (equivalent to 0.6 mg or 300 .mu.M
colchicine) vial of 30 tablets were tested. 10 mL of sterile water
was added for injection.
[0112] First, the cover from the colchicine vial was removed. The
contents of 2 tablets were crushed into a fine powder. Then, 10 mL
of sterile water was added, for injection, to the vial. The
solution was then shaken well and sonicated for 10 minutes or until
all of the powder was dissolved. Next, the solution was centrifuged
for 10 minutes. The supernatant was retained and was diluted to the
final assay concentration.
Colchicine Capsules+Sterile Water for Injection
[0113] Colchicine capsules (equivalent to 0.6 mg or 300 .mu.M
colchicine), vial of 100 capsules, were tested. 10 mL of sterile
water was added for injection.
[0114] First, the cover from Colchicine vial was removed. 2
capsules were then carefully opened, and the contents were emptied
into a vial. 10 mL of sterile water was added for injection to the
vial. The well was then shaken and sonicated for 10 minutes or
until all of the powder was dissolved. The product was then
centrifuged for 10 minutes. The supernatant was retained and was
diluted to the final assay concentration.
Methods
Mass Spectrometry Method Development
[0115] The signal was optimized for each compound by ESI positive
or negative ionization mode. An MS2 scan or an SIM scan was used to
optimize the fragmenter voltage and a product ion analysis was used
to identify the best fragment for analysis, and the collision
energy was optimized using a product ion or MRM scan. An ionization
ranking was assigned indicating the compound's ease of
ionization.
Analysis
[0116] Samples were analyzed by LC/MS/MS using an Agilent 6410 mass
spectrometer coupled with an Agilent 1200 HPLC and a CTC PAL
chilled autosampler, all controlled by MassHunter software
(Agilent). After separation on a C18 reverse phase HPLC column
(Agilent Zorbax 3.5 um, 2.1.times.30 mm) using an
acetonitrile-water gradient, peaks were analyzed by mass
spectrometry using ESI ionization in MRM mode; Solution A contained
H.sub.2O with 0.1% formic acid, and Solution B contained
acetonitrile with 0.1% formic acid. Tables 12 and 13 show data
regarding HPLC gradient and experimental conditions.
TABLE-US-00013 TABLE 12 HPLC Gradient Time Flow rate % A % B (min)
(mL/min) Mobile Phase Mobile Phase 0.37 1 98 2 1.88 1 5 95 2.06 1 5
95 2.14 1 98 2 3 1 98 2
TABLE-US-00014 TABLE 13 Caco-2 Permeability: Experimental
Conditions Test Test Transport Reference Analytical Article conc.
pH Direction compounds method Colchicine-Liquid 10 .mu.M 5.7/7.4
A.fwdarw.B ranitidine LC/MS/ Colchicine, USP- 6.5/7.4 B.fwdarw.A
talinolol MS Tablets 7.4/7.4 metoprolol Colchicine- Capsules
Summary of Procedure:
[0117] Caco-2 cells were grown in tissue culture flasks and were
trypsinized, suspended in medium, and the suspensions were applied
to wells of a Millipore 96 well Caco-2 plate. The cells were
allowed to grow and differentiate for three weeks, and fed at 2-day
intervals.
[0118] For Apical to Basolateral (A.fwdarw.B) permeability, the
test agent was added to the apical (A) side and amount of
permeation was determined on the basolateral (B) side; for
Basolateral to Apical (B.fwdarw.A) permeability, the test agent was
added to the B side and the amount of permeation was determined on
the A side. The A-side buffer contained 100 .mu.M Lucifer yellow
dye, in Transport Buffer (1.98 g/L glucose in 10 mM HEPES, lx
Hank's Balanced Salt Solution) at pH 5.7, 6.5, or 7.4, and the
B-side buffer used was Transport Buffer, pH 7.4. Caco-2 cells were
incubated with these buffers for 2 hours, and the receiver side
buffer was removed for analysis by LC-MS/MS.
[0119] To verify the Caco-2 cell monolayers were properly formed,
aliquots of the cell buffers were analyzed by fluorescence to
determine the transport of the impermeable dye Lucifer Yellow. Any
deviations from control values were reported.
Data expressed as permeability (P.sub.app):
P.sub.app=dQ/Dt/C.sub.0A.
dQ/dt: rate of permeation; C.sub.0: initial concentration of test
agent; A: area of monolayer. For bidirectional permeability
studies, Efflux Ratio (RE):
R.sub.e=P.sub.app(B.fwdarw.A)/P.sub.app(A.fwdarw.B). Table 14 shows
mass spectrometry method development. Tables 15 and 16 show data
regarding Caco-2 permeability data displaying a data summary and
individual data from replicates, respectively.
Results
TABLE-US-00015 [0120] TABLE 14 Mass Spectrometry Method
Development: MS/MS Test Mono- ESI Article isotopic Polar- Precursor
Product Ionization (API) Mass ization m/2z m/z classification
Colchicine 399.17 Positive 399.9 358.2 1 Ionization classification:
1 = Highly ionizable; 2 = Intermediately ionizable; 3 = Poorly
ionizable m/z: Mass-to-Charge (or, mass-to-double charge) ratio of
analyte
TABLE-US-00016 TABLE 15 Caco-2 Permeability: Data Summary mean mean
A.fwdarw.B B.fwdarw.A Papp Papp Ef- Test Test pH (10-6 (10-6 flux
Article conc. (A/B) cm s-1) cm s-1) ratio Comment ranitidine 10
.mu.M 6.5/7.4 0.27 2.1 7.8 Class III BCS talinolol 10 .mu.M 6.5/7.4
0.073 6.6 90.4 P-gp efflux control metoprolol 10 .mu.M 6.5/7.4 13.7
27.2 2.0 high BCS control (Class I BCS) Colchicine- 10 .mu.M
5.7/7.4 0.10 5.4 54.0 Liquid Colchicine, 10 .mu.M 5.7/7.4 0.12 4.9
40.8 USP- Tablets Colchicine- 10 .mu.M 5.7/7.4 0.15 5.2 34.7
Capsules Colchicine- 10 .mu.M 6.5/7.4 0.16 5.4 33.8 Liquid
Colchicine, 10 .mu.M 6.5/7.4 0.17 5.2 30.6 USP- Tablets Colchicine-
10 .mu.M 6.5/7.4 0.14 5.3 37.9 Capsules Colchicine- 10 .mu.M
7.4/7.4 0.12 5.8 48.3 Liquid Colchicine, 10 .mu.M 7.4/7.4 0.13 5.8
44.6 USP-Tablet Colchicine- 10 .mu.M 7.4/7.4 0.18 6.4 35.6
Capsules
TABLE-US-00017 TABLE 16 Caco-2 Permeability: Individual Data from
Replicates Trans- Papp (10.sub.-6 cm s-1) A.fwdarw.B port 1.sup.st
2.sup.nd 3.sup.rd Post- Test pH Direc- repli- repli- repli- Assay
Article (A/B) tion cate cate cate Mean Recovery Colchicine- 5.7/7.4
A.fwdarw.B 0.10 0.10 0.10 0.10 61% Liquid B.fwdarw.A 5.2 5.2 5.4
5.1 6.5/7.4 A.fwdarw.B 0.19 0.13 0.16 0.16 63% B.fwdarw.A 5.6 5.1
5.5 5.4 7.4/7.4 A.fwdarw.B 0.13 0.13 0.091 0.12 64% B.fwdarw.A 5.3
6.1 6.1 5.7 Colchicine, 5.7/7.4 A.fwdarw.B 0.15 0.11 0.091 0.12 65%
USP- B.fwdarw.A 4.7 5.1 5.0 4.9 Tablets 6.5/7.4 A.fwdarw.B 0.12
0.13 0.25 0.17 63% B.fwdarw.A 4.7 5.4 5.6 5.1 7.4/7.4 A.fwdarw.B
0.16 0.15 0.093 0.13 63% B.fwdarw.A 5.5 5.7 6.3 5.6 Colchicine-
5.7/7.4 A.fwdarw.B 0.15 0.17 0.13 0.15 63% Capsules B.fwdarw.A 5.0
5.1 5.4 5.0 6.5/7.4 A.fwdarw.B 0.18 0.084 0.15 0.14 64% B.fwdarw.A
5.2 5.1 5.5 5.1 7.4/7.4 A.fwdarw.B 0.16 0.19 0.19 0.18 63%
B.fwdarw.A 6.3 6.2 6.6 6.2 P.sub.app: apparent permeability rate
coefficient
QC Criteria
[0121] All wells utilized in this study passed QC criteria for
epithelial monolayer integrity. All wells utilized exhibited
Lucifer Yellow transport less than 2% (unless noted otherwise). All
wells that were utilized in this assay exhibited TEER
(Trans-Epithelial Electrical Resistance) readings greater than 1500
Ohms, also in accordance with QC criteria for acceptable monolayer
integrity. FIG. 3 shows the calibration curve of colchicine in
Caco-2 transport buffer.
[0122] BCS criterion classifies all 3 batches of Colchicine tested
in this study as exhibiting low permeability across the pH ranges
tested (5.7, 6.5, and 7.4). All 3 batches of Colchicine tested in
this study (USP tablets, capsules, and liquid) exhibited a very
similar permeability profile (i.e. low permeability with high
efflux ratios and similar post-assay recovery levels).
[0123] The apparent permeability rate coefficients (P.sub.app) for
all 3 batches of Colchicine tested in this study (in the
absorptive, A.fwdarw.B, transport direction) approximates only 1%
(or less) of the Papp coefficient of the BCS high permeability
control compound, metoprolol.
[0124] This value of 1% does not meet the 90% threshold of FDA
criteria for BCS classification as high permeability. As a result,
all 3 batches of Colchicine tested in this study are classified as
exhibiting low permeability for BCS purposes. (Current EMA
guidelines for BCS classification are 80% of the positive control
for high permeability). FIG. 4 shows a colchicine chromatograms in
Caco-2 buffer. Table 17 shows the MRM transition for colchicine.
Table 18 shows cumulative colchicine oral solution data at
25.degree. C..+-.2.degree. C./60%.+-.5% RH, which has been stable
for at least 3 months. Table 19 shows the cumulative colchicine
oral solution data at 40.degree. C..+-.2.degree. C./75%.+-.5% RH,
which has been stable for at least 3 months. It is mandatory to add
a suitable preservative to non-sterile solutions, it maintain
microbiological integrity for the remainder of the shelf-life. The
preservative should also be stable to maintain its anti-microbial
activity. Benzyl alcohol, a preservative in the colchicine oral
solution tested in Tables 18 and 19, is stable and is compatible
with colchicine as well as all excipients.
[0125] The data presented herein also shows that over a period of
time the active drug-substance (colchicine) does not degrade more
than the accepted limit and the each of the degradants does not
exceed a certain threshold. No such information is available in the
literature for liquid colchicine. As you can see, the data herein
show that, fortunately, the degradants, after 3-month storage, even
at 40.degree. C./75% RH (indicative of stability to 24-month at
room temperature), are either not detected or not quantifiable
(very small quantity).
[0126] A liquid formulation of colchicine has been made for oral
consumption which is stable, has a compatible and stable
preservative, and under degrades minimally in longer-term storage
conditions (well below the acceptable limit).
TABLE-US-00018 TABLE 17 MRM Transition for Colchicine: MS/MS Method
Development Mono- ESI Test Isotopic Polar- Precursor Product
Ionization Article Mass ization m/z m/z Classification Colchicine
399.17 Positive 399.9 358.2 1 m/z: mass-to-charge ratio of analyte
Ionization Classification: 1 = Highly Ionizable
TABLE-US-00019 TABLE 18 Cumulative Colchicine Oral Solution Data at
25.degree. C. .+-. 2.degree. C./60% .+-. 5% RH Stability Data:
Colchicine Oral Solution Batch No.: PD16033 Storage Condition:
25.degree. .+-. 2.degree. C./60% .+-. 5% RH Drug Substance Batch
No.: 31626/H Container Closure: 190-cc, Oblong, HDPE Bottles with
38-400 mm Auto-Loc CRC with a foil liner Test Parameter Analytical
Acceptance Results [Storage (Months)] Procedure Criteria 0 1 2 3 6
Description A slightly Con- Con- Con- Con- Organoleptic hazy, red
forms forms forms forms liquid with a cherry odor. pH 6.0 to 6.6
6.6 6.6 6.6 USP <791> 7.2 Density 1.00 to 1.02 1.02 1.02 1.02
USP <841> 1.04 g/mL at 25.degree. C. Colchicine 0.108 to 99.3
100.0 100.0 99.8 Assay 0.132 mg/mL In-house HPLC (90.0 to 110.0% of
label claim) Benzyl Alcohol 2.4 to 99 99 98 98 Assay 3.6 mg/mL
In-house HPLC (80 to 120% of label claim) Related Substances
In-house HPLC .beta.- NMT 1.0% NT NT NT ND lumicolchicine .gamma.-
NMT 1.0% NT NT NT ND lumicolchicine Colchiceine NMT 1.0% NT NT NT
<LOQ Any other NMT 1.0% NT NT NT ND individual unknown
impurities Total NMT 5.0% NT NT NT <LOQ Impurities HPLC: High
Performance Liquid Chromatography NMT: Not More Than NLT: Not Less
Than NT: Not Tested (Testing not required by development
specification.)
TABLE-US-00020 TABLE 19 Cumulative Colchicine Oral Solution Data at
40.degree. C. .+-. 2.degree. C./75% .+-. 5% RH Stability Data:
Colchicine Oral Solution Batch No.: PD16033 Storage Condition:
40.degree. .+-. 2.degree. C./75% .+-. 5% RH Drug Substance Batch
No.: 31626/H Container Closure: 190-cc, Oblong, HDPE Bottles with
38-400 mm Auto-Loc CRC with a foil liner Test Parameter Analytical
Acceptance Results [Storage (Months)] Procedure Criteria 0 1 2 3 6
Description A slightly Con- Con- Con- Con- Organoleptic hazy, red
forms forms forms forms liquid with a cherry odor. pH 6.0 to 6.6
6.6 6.6 6.6 USP <791> 7.2 Density 1.00 to 1.02 1.02 1.02 1.02
USP <841> 1.04 g/mL at 25.degree. C. Colchicine 0.108 to 99.3
99.2 99.6 97.4 Assay 0.132 mg/mL In-house HPLC (90.0 to 110.0% of
label claim) Benzyl Alcohol 2.4 to 99 97 96 93 Assay 3.6 mg/mL
In-house HPLC (80 to 120% of label claim) Related Substances
In-house HPLC .beta.- NMT 1.0% NT NT NT ND lumicolchicine .gamma.-
NMT 1.0% NT NT NT ND lumicolchicine Colchiceine NMT 1.0% NT NT NT
<LOQ Any other NMT 1.0% NT NT NT ND individual unknown
impurities Total NMT 5.0% NT NT NT <LOQ Impurities HPLC: High
Performance Liquid Chromatography NMT: Not More Than NLT: Not Less
Than NT: Not Tested (Testing not required by development
specification.)
REFERENCES
[0127] Habib, M. J. et al (2008). "Influence of certain additives
on the photostability of colchicine solutions." Drug Development
and Industrial Pharm. 15(11):1905-1909. [0128] Stewart, B. H. et
al. (1995). "Comparison of Intestinal Permeabilities Determined in
Multiple In Vitro and In Situ Models: Relationship to Absorption in
Humans." Pharm. Res. 12:693-699. [0129] Artursson, P. et al.
(2001). "Caco-2 Monolayers in Experimental and Theoretical
Predictions of Drug Transport." Adv. Drug Deliv. Rev. 46:27-43.
[0130] Yee, Shiyin (1997). "In Vitro Permeability across Caco-2
Cells (Colonic) Can Predict In Vivo (Small Intestinal) Absorption
in Man--Fact or Myth." Pharm. Res 14(6):763-766. [0131] Yu, L. X.
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[0132] The foregoing written specification is considered to be
sufficient to enable one skilled in the art to practice the
invention. The present invention is not to be limited in scope by
examples provided, since the examples are intended as a single
illustration of one aspect of the invention and other functionally
equivalent embodiments are within the scope of the invention.
Various modifications of the invention in addition to those shown
and described herein will become apparent to those skilled in the
art from the foregoing description and fall within the scope of the
appended claims. The advantages and objects of the invention are
not necessarily encompassed by each embodiment of the
invention.
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