U.S. patent application number 11/927011 was filed with the patent office on 2008-07-03 for sustained release formulation of a peptide.
Invention is credited to Jacques-Pierre Moreau.
Application Number | 20080161233 11/927011 |
Document ID | / |
Family ID | 34525703 |
Filed Date | 2008-07-03 |
United States Patent
Application |
20080161233 |
Kind Code |
A1 |
Moreau; Jacques-Pierre |
July 3, 2008 |
Sustained Release Formulation of a Peptide
Abstract
This invention pertains to a sustained release complex, Compound
(I), which comprises Compound (A), having the formula ##STR00001##
or a pharmaceutically acceptable salt thereof, and a copolymer
comprising poly-(I)-lactic-glycolic-tartaric acid (P(I)LGT),
wherein the amino group of said Compound (A) is ionically bound to
a carboxyl group of the P(I)LGT. The present invention further
pertains to a process for making said sustained release complex.
Further still, the present invention is directed to a
pharmaceutical composition comprising said sustained release
complex and a pharmaceutically acceptable carrier(s).
Inventors: |
Moreau; Jacques-Pierre;
(Upton, MA) |
Correspondence
Address: |
Biomeasure, Incorporated
27 Maple Street
Milford
MA
01757-3650
US
|
Family ID: |
34525703 |
Appl. No.: |
11/927011 |
Filed: |
October 29, 2007 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10990033 |
Nov 16, 2004 |
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11927011 |
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10048856 |
May 15, 2002 |
7109166 |
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PCT/US00/22464 |
Aug 16, 2000 |
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10990033 |
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60149649 |
Aug 18, 1999 |
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Current U.S.
Class: |
424/426 ;
514/15.7; 514/18.2; 514/19.3; 514/6.9 |
Current CPC
Class: |
A61P 1/12 20180101; Y10S
514/806 20130101; Y10S 514/964 20130101; A61P 1/00 20180101; A61P
17/06 20180101; A61P 35/02 20180101; A61K 47/593 20170801; Y10S
514/951 20130101; Y10S 514/908 20130101; Y10S 514/867 20130101 |
Class at
Publication: |
514/11 |
International
Class: |
A61K 38/12 20060101
A61K038/12; A61P 17/06 20060101 A61P017/06; A61P 1/00 20060101
A61P001/00 |
Claims
1-8. (canceled)
9. A method of treating a disease or condition in a patient in need
thereof, which comprises administering to said patient an effective
amount of Compound (A), ##STR00006## or a pharmaceutically
acceptable salt thereof, wherein the disease or condition is
selected from the group consisting of systemic sclerosis,
pancreatic pseudocysts, pancreatic ascites, VIPoma,
nesidoblastosis, hyperinsulinism, gastrinoma, Zollinger-Ellison
Syndrome, hypersecretory diarrhea, scleroderma, irritable bowel
syndrome, upper gastrointestinal bleeding, postprandial portal
venous hypertension, complications of portal hypertension, small
bowel obstruction, duodenogastric reflux, Cushing's Syndrome,
gonadotropinoma, hyperparathyroidism, diabetic neuropathy,
hypercalcemia of malignancy, Paget's disease, meningioma, cancer
cachexia, psoriasis, hypotension and panic attacks.
10. A method of treating a disease or condition in a patient in
need thereof, which comprises administering to said patient an
effective amount of Compound (I), which comprises Compound (A)
having the formula ##STR00007## and a polymer, wherein the polymer
comprises lactide units, glycolide units and tartaric acid units
where the ratio in the polymer of the lactide units is from and
including about 71% to about 73%, of the glycolide units is from
and including about 26% to about 28%; and of the tartaric acid
units is from and including about 1% to about 3%; and where the
amino group of Compound (A) is ionically bonded to a carboxylic
group of the acid units of the polymer, and wherein the disease or
condition is selected from the group consisting of systemic
sclerosis, pancreatic pseudocysts, pancreatic ascites, VIPoma,
nesidoblastosis, hyperinsulinism, gastrinoma, Zollinger-Ellison
Syndrome, hypersecretory diarrhea, scleroderma, irritable bowel
syndrome, upper gastrointestinal bleeding, postprandial portal
venous hypertension, complications of portal hypertension, small
bowel obstruction, duodenogastric reflux, Cushing's Syndrome,
gonadotropinoma, hyperparathyroidism, diabetic neuropathy,
hypercalcemia of malignancy, Paget's disease, meningioma, cancer
cachexia, psoriasis, hypotension and panic attacks.
11. A method of treating a disease or condition in a patient in
need thereof, which comprises administering to said patient an
effective amount of Compound (I), which comprises Compound (A)
having the formula ##STR00008## and a polymer, wherein the polymer
comprises lactide units, glycolide units and tartaric acid units
where the ratio in the polymer of the lactide units is from and
including about 71% to about 73%, of the glycolide units is from
and including about 26% to about 28%; and of the tartaric acid
units is from and including about 1% to about 3%; and where the
amino group of Compound (A) is ionically bonded to a carboxylic
group of the acid units of the polymer, and wherein the Compound
(I) is in the form of microparticles, and 4, wherein the disease or
condition is selected from the group consisting of systemic
sclerosis, pancreatic pseudocysts, pancreatic ascites, VIPoma,
nesidoblastosis, hyperinsulinism, gastrinoma, Zollinger-Ellison
Syndrome, hypersecretory diarrhea, scleroderma, irritable bowel
syndrome, upper gastrointestinal bleeding, postprandial portal
venous hypertension, complications of portal hypertension, small
bowel obstruction, duodenogastric reflux, Cushing's Syndrome,
gonadotropinoma, hyperparathyroidism, diabetic neuropathy,
hypercalcemia of malignancy, Paget's disease, meningioma, cancer
cachexia, psoriasis, hypotension and panic attacks.
Description
BACKGROUND OF THE INVENTION
[0001] This invention pertains to a sustained release complex,
Compound (I), which comprises Compound (A), having the formula
##STR00002##
or a pharmaceutically acceptable salt thereof, and a copolymer
comprising poly-(1)-lactic-glycolic-tartaric acid (P(I)LGT),
wherein the amino group of said Compound (A) is ionically bound to
a carboxyl, group of the P(I)LGT. The present invention further
pertains to a process for making said sustained release complex.
Further still, the present invention is directed to a
pharmaceutical composition comprising said sustained release
complex and a pharmaceutically acceptable carrier(s).
[0002] Further, since Compound (A) is an analogue of somatostatin
and it is well known to those skilled in the art that the known and
potential uses of somatostatin are varied and multitudinous, this
invention is also directed to the use of Compound (A), Compound (I)
or microparticles of Compound (I) to treat a disease or condition
in a patient in need thereof, which comprises administering
Compound (A), Compound (I) or microparticles of Compound (I) to
said patient, wherein the diseases or conditions to be treated are
selected from the group consisting of gastroenterological
conditions and/or diseases, such as Crohn's disease, systemic
sclerosis, external and internal pancreatic pseudocysts and
ascites, VIPoma, nesidoblastosis, hyperinsulinism, gastrinoma,
Zollinger-Ellison Syndrome, diarrhea, AIDS related diarrhea,
chemotherapy related diarrhea, scleroderma, Irritable Bowel
Syndrome, pancreatitis, upper gastrointestinal bleeding,
postprandial portal venous hypertension especially in cirrhotic
patients, complications of portal hypertension, small bowel
obstruction, gastroesophageal reflux, duodenogastric reflux and in
treating endocrinological diseases and/or conditions, such as
Cushing's Syndrome, gonadotropinoma, hyperparathyroidism, Graves'
Disease, diabetic neuropathy, macular degeneration, hypercalcemia
of malignancy, Paget's disease, and polycystic ovary disease; in
treating various types of cancer such as thyroid cancer, leukemia,
meningioma and conditions associated with cancer such as cancer
cachexia; in the treatment of such conditions as hypotension such
as orthostatic hypotension and postprandial hypotension and panic
attacks.
[0003] Many drug delivery systems have been developed tested and
utilized for the controlled in vivo release of pharmaceutical
compositions. For example, polyesters such as poly(DL-lactic acid),
poly(glycolic acid), poly(.epsilon.-caprolactone) and various other
copolymers have been used to release biologically active molecules
such as progesterone; these have been in the form of microcapsules,
films or rods (M. Chasin and R. Langer, editors, Biodegradable
Polymers as Drug Delivery Systems, Dekker, N Y 1990). Upon
implantation of the polymer/therapeutic agent composition, for
example, subcutaneously or intramuscularly, the therapeutic agent
is released over a specific period of time. Such bio-compatible
biodegradable polymeric systems are designed to permit the
entrapped therapeutic agent to diffuse from the polymer matrix.
Upon release of the therapeutic agent, the poller is degraded in
vivo, obviating surgical removal of the implant. Although the
factors that contribute to poller degradation are not well
understood, it is believed that such degradation for polyesters may
be regulated by the accessibility of ester linkages to
non-enzymatic autocatalytic hydrolysis of the polymeric
components.
[0004] Several EPO publications and U.S. patents have addressed
issues of polymer matrix design and its role in regulating the rate
and extent of release of therapeutic agents in vivo.
[0005] For example, Deluca (EPO Publication 0 467 389 A2) describes
a physical interaction between a hydrophobic biodegradable polymer
and a protein or polypeptide. The composition formed was a mixture
of a therapeutic agent and a hydrophobic polymer that sustained its
diffusional release from the matrix after introduction into a
subject.
[0006] Hutchinson (U.S. Pat. No. 4,767,628) controlled the release
of a therapeutic agent by uniform dispersion in a polymeric device.
It is disclosed that this formulation provides for controlled
continuous release by the overlap of two phases: first, a
diffusion-dependent leaching of the drug from the surface of the
formulation; and second, releasing by aqueous channels induced by
degradation of the polymer.
[0007] PCT publication WO 93/24150 discloses a sustained release
formulation comprising a peptide having a basic group and a
carboxy-terminated polyester.
[0008] U.S. Pat. No. 5,612,052 describes cation-exchanging
microparticles made typically of carboxyl-bearing polyester chains
onto which basic bioactive agents are immobilized to provide a
control release system within an absorbable gel-forming liquid
polyester.
[0009] Compound (A) is described and claimed in U.S. Pat. No.
5,552,520, which is assigned to the assignee hereof.
[0010] PCT publication WO 97/40085, assigned to the assignee
hereof, discloses biodegradable polyesters comprising lactic acid
units, glycolic acid units and hydroxy-polycarboxylic acid units
such as tartaric acid or pamoic acid and processes for making said
polyesters. More specifically, it discloses
poly-lactide-glycolide-tartaric acid polymers in the ratio 65/33/2,
respectively.
[0011] PCT publication WO 94/15587, assigned to the assignee
hereof, discloses ionic conjugates of polyesters having free COOH
groups with a bioactive peptide having at least one effective
ionogenic amine. More specifically, it discloses that the polymers
are made polycarboxylic by reacting the co-polymers with malic acid
or citric acid. U.S. Pat. No. 5,672,659, is the U.S. national phase
continuation application of WO 94/15587. U.S. Pat. No. 5,863,985 is
a continuation of U.S. Pat. No. 5,672,659. Pending U.S. application
Ser. No. 09/237,405 is a CIP of U.S. Pat. No. 5,863,985, which
additionally discloses a polyester which must include citric acid,
.epsilon.-caprolactone and glycolide; compositions comprising the
immediately foregoing polyesters and a polypeptide; a polyester
that must include tartaric acid as one of its members; compositions
comprising the immediately foregoing polyester and a polypeptide;
and the foregoing compositions in the shape of rods which are
optionally coated with a biodegradable polymer.
[0012] PCT publication WO 97/39738, assigned to the assignee
hereof, discloses a method of making microparticles of a sustained
release ionic conjugate as described in WO 94/15587.
[0013] The contents of the foregoing patents, applications and
publications are incorporated herein in their entirety.
[0014] The present invention is directed to a preferred embodiment
of a sustained release ionic conjugate of polymer
poly-lactide-glycolide-tartaric acid and Compound (A), also known
as Compound (I), which is characterized by the surprising and
non-obvious property of zero-order release of Compound (A) from the
conjugate. More preferably, the ionic conjugate, Compound (I), is
in the form of microparticles.
BRIEF DESCRIPTION OF DRAWINGS
[0015] FIG. 1: Shows the in vivo release profile of Compound (A)
from a sample of Compound (I) in dog, wherein the sample of
Compound (I) consists of about 11.23% Compound (A), the polymer is
I-lactide:glycolide:tartaric acid (72:27:1) and where Compound (I)
is administered intramuscularly as microparticles. The irradiated
sample refers to a sample of Compound (I) which was irradiated with
.gamma.-rays from a Cobalt source.
SUMMARY OF THE INVENTION
[0016] The present invention is directed to a Compound (I) which
comprises Compound (A), having the formula
##STR00003##
and a polymer, wherein the polymer comprises lactide units,
glycolide units and tartaric acid units where the ratio in the
polymer: of the lactide units is from and including about 71% to
about 73%, of the glycolide units is from and including about 26%
to about 28%; and of the tartaric acid units is from and including
about 1% to about 3%; and where the amino group of Compound (A) is
ionically bonded to a carboxylic group of the acid units of the
polymer.
[0017] A preferred embodiment of Compound (I) is where the polymer
consists of about 72% lactide units, about 27% glycolide units and
about 1% tartaric acid units.
[0018] A preferred embodiment of the immediately foregoing Compound
(I) is where the percentage of Compound (A) in Compound (I) is
about 8% to about 12%.
[0019] In another aspect, the present invention is directed to
microparticles of Compound (I) which comprises Compound (A), having
the formula
##STR00004##
and a polymer, wherein the polymer comprises lactide units,
glycolide units and tartaric acid units where the ratio in the
polymer: of the lactide units is from and including about 71% to
about 73%, of the glycolide units is from and including about 26%
to about 28%; and of the tartaric acid units is from and including
about 1% to about 3%; and where the amino group of Compound (A) is
ionically bonded to a carboxylic group of the acid units of the
polymer.
[0020] Preferred microparticles of Compound (I), as described
hereinabove, of this invention are those microparticles having a
mean microparticle size of about 10 microns to about 100
microns.
[0021] More preferred microparticles of Compound (I), as described
hereinabove, of this invention are those microparticles having a
mean microparticle size of about 40 microns to about 70
microns.
[0022] Even more preferred microparticles of the present invention
is where the microparticles exhibit a zero-order release profile of
Compound (A) from the microparticles.
[0023] In yet another aspect, the present invention is directed to
a pharmaceutical composition comprising microparticles comprising
Compound (I) which comprises Compound (A), having the formula:
##STR00005##
and a polymer, wherein the polymer comprises lactide units,
glycolide units and tartaric acid units where the ratio in the
polymer: of the lactide units is from and including about 71% to
about 73%, of the glycolide units is from and including about 26%
to about 28%; and of the tartaric acid units is from and including
about 1% to about 3%; and where the amino group of Compound (A) is
ionically bonded to a carboxylic group of the acid units of the
polymer; and optionally a pharmaceutically acceptable carrier,
diluent or adjuvant.
[0024] In a further aspect, the present invention is directed to a
method of treating a disease or condition in a patient in need
thereof, which comprises administering to said patient an effective
amount, of Compound (A), as described hereinabove, or a
pharmaceutically acceptable salt thereof, wherein the disease or
condition is selected from the group consisting of systemic
sclerosis, pancreatic pseudocysts, pancreatic ascites, VIPoma,
nesidoblastosis, hyperinsulinism, gastrinoma, Zollinger-Ellison
Syndrome, hypersecretory diarrhea, scleroderma, irritable bowel
syndrome, upper gastrointestinal bleeding, postprandial portal
venous hypertension, complications of portal hypertension, small
bowel obstruction, duodenogastric reflux, Cushing's Syndrome,
gonadotropinoma, hyperparathyroidism, diabetic neuropathy, macular
degeneration, hypercalcemia of malignancy, Paget's disease,
meningioma, cancer cachexia, psoriasis, hypotension and panic
attacks.
[0025] In an even further aspect, the present invention is directed
to a method of treating a disease or condition in a patient in need
thereof, which comprises administering to said patient an effective
amount of Compound (I), as described hereinabove, wherein the
disease or condition is selected from the group consisting of
systemic sclerosis, pancreatic pseudocysts, pancreatic ascites,
VIPoma, nesidoblastosis, hyperinsulinism, gastrinoma,
Zollinger-Ellison Syndrome, hypersecretory diarrhea, scleroderma,
irritable bowel syndrome, upper gastrointestinal bleeding,
postprandial portal venous hypertension, complications of portal
hypertension, small bowel obstruction, duodenogastric reflux,
Cushing's Syndrome, gonadotropinoma, hyperparathyroidism, diabetic
neuropathy, macular degeneration, hypercalcemia of malignancy,
Paget's disease, meningioma, cancer cachexia, psoriasis,
hypotension and panic attacks.
[0026] In still a further aspect, the present invention is directed
to a method of treating a disease or condition in a patient in need
thereof, which comprises administering to said patient an effective
amount of microparticles of Compound (I), as described hereinabove,
wherein the disease or condition is selected from the group
consisting of systemic sclerosis, pancreatic pseudocysts,
pancreatic ascites, VIPoma, nesidoblastosis, hyperinsulinism,
gastrinoma, Zollinger-Ellison Syndrome, hypersecretory diarrhea,
scleroderma, irritable bowel syndrome, upper gastrointestinal
bleeding, postprandial portal venous hypertension, complications of
portal hypertension, small bowel obstruction, duodenogastric
reflux, Cushing's Syndrome, gonadotropinoma, hyperparathyroidism,
diabetic neuropathy, macular degeneration, hypercalcemia of
malignancy, Paget's disease, meningioma, cancer cachexia,
psoriasis, hypotension and panic attacks.
DETAILED DESCRIPTION
[0027] The term "about" as used herein in association with
parameters and amounts, means that the parameter or amount is
within .+-.5% of the stated parameter or amount.
[0028] The term "microparticle(s)" as used herein, refers to the
micron size particles of the ionic conjugate comprising Compound
(A) and poly-lactide-glycolide-tartaric acid polymer, which are
preferably in essentially spherical form.
[0029] The instant application denotes amino acids using the
standard three letter abbreviation known in the art, for example
Phe=phenylalanine; Abu=.alpha.-aminobutyric acid.
[0030] As is well known to those skilled in the art, the known and
potential uses of somatostatin are varied and multitudinous.
Somatostatin is known to be useful in the treatment of the diseases
and/or conditions listed hereinbelow. The varied uses of
somatostatin may be summarized as follows: Cushings Syndrome (see
Clark, R. V. et al, Clin. Res. 38, p. 943A, 1990); gonadotropinoma
(see Ambrosi B., et al., Acta Endocr. (Copenh.) 122, 569-576,
1990); hyperparathyroidism (see Miller, D., et al., Canad. Med.
Ass. J., Vol. 145, pp. 227-228, 1991); Paget's disease (see,
Palmieri, G. M. A., et al., J. of Bone and Mineral Research, 7,
(Suppl. 1), p. S240 (Abs. 591), 1992); VIPoma (see Koberstein, B.,
et al., Z. Gastroenterology, 28, 295-301, 1990 and Christensen, C.,
Acta Chir. Scand. 155, 541-543, 1989); nesidioblastosis and
hyperinsulinism (see Laron, Z., Israel J. Med. Sci., 26, No. 1,
1-2, 1990, Wilson, D.C., Irish J. Med. Sci., 158, No. 1, 31-32,
1989 and Micic, D., et al., Digestion, 16, Suppl. 1.70. Abs. 193,
1990); gastrinoma (see Bauer, F. E., et al., Europ. J. Pharmacol.,
183, 55 1990); Zollinger-Ellison Syndrome (see Mozell, E., et al.,
Surg. Gynec. Obstet., 170, 476-484, 1990); hypersecretory diarrhea
related to AIDS and other conditions (due to AIDS, see Cello, J.
P., et al., Gastroenterology, 98, No. 5, Part 2, Suppl., A163 1990;
due to elevated gastrin-releasing peptide, see Alhindawi, R., et
al., Can. J. Surg., 33, 139-142, 1990; secondary to intestinal
graft vs. host disease, see Bianco J. A., et al., Transplantation,
49, 1194-1195, 1990; diarrhea associated with chemotherapy, see
Petrelli, N., et al., Proc. Amer. Soc. Clin. Oncol., Vol. 10, P
138, Abstr. No. 417 1991); irritable bowel syndrome (see O'Donnell,
L. J. D., et al., Aliment. Pharmacol. Therap., Vol. 4, 177-181,
1990); pancreatitis (see Tulassay, Z., et al., Gastroenterology,
98, No. 5, Part 2, Suppl., A238, 1990); Crohn's Disease (see
Fedorak, R. N., et al., Can. J. Gastroenterology, 3, No. 2, 53-57,
1989); systemic sclerosis (see Soudah, H., et al.,
Gastroenterology, 98, No. 5, Part 2. Suppl., A129, 1990); thyroid
cancer (see Modigliani, E., et al., Ann., Endocr. (Paris), 50,
483-488, 1989); psoriasis (see Camisa, C., et al., Cleveland Clinic
J. Med., 57, No. 1, 71-76, 1990); hypotension (see Hoeldtke, R. D.,
et al., Arch. Phys. Med. Rehabil., 69, 895-898, 1988 and Kooner, J.
S., et al., Brit. J. Clin. Pharmacol., 28, 735P-736P, 1989); panic
attacks (see Abelson, J. L., et al., Clin. Psychopharmacol., 10,
128-132, 1990); sclerodoma (see Soudah, H., et al., Clin. Res.,
Vol. 39, p. 303A, 1991); small bowel obstruction (see Nott, D. M.,
et al., Brit. J. Surg., Vol. 77, p. A691, 1990); gastroesophageal
reflux (see Branch, M. S., et al., Gastroenterology, Vol. 100, No.
5, Part 2 Suppl., p. A425, 1991); duodenogastric reflux (see
Hasler, W., et al., Gastroenterology, Vol. 100, No. 5, Part 2,
Suppl., p. A448, 1991); Graves' Disease (see Chang, T. C., et al.,
Brit. Med. J., 304, p. 158, 1992); polycystic ovary disease (see
Prelevic, G. M., et al., Metabolism Clinical and Experimental, 41,
Suppl. 2, pp 76-79, 1992); upper gastrointestinal bleeding (see
Jenkins, S. A., et al., Gut., 33, pp. 404-407, 1992 and Arrigoni,
A., et al., American Journal of Gastroenterology, 87, p. 1311,
(abs. 275), 1992); pancreatic pseudocysts and ascites (see.
Hartley, J. E., et al., J. Roy. Soc. Med., 85, pp. 107-108, 1992);
leukemia (see Santini, et al., 78, (Suppl. 1), p. 429A (Abs. 1708),
1991); meningioma (see Koper, J. W., et al., J. Clin. Endocr.
Metab., 74, pp. 543-547, 1992); and cancer cachexia (see Bartlett,
D. L., et al., Surg. Forum., 42, pp. 14-16, 1991). The contents of
the foregoing references are incorporated herein by reference.
[0031] Applicant has now discovered that Compound (A), which is a
somatostatin agonist, Compound (I) and microparticles of Compound
(I), are particularly useful in treating the conditions, disorders
and diseases noted hereinabove.
General Procedures:
[0032] Co-Polymer formation: The co-polymer consisting of
L-lactide, glycolide and L(+)-tartaric acid can be made according
to methods well-known to those skilled in the art and as enabled
herein. Accordingly, a reactor is loaded with monomers of
glycolide, L-lactide and L(+)-tartaric acid and stannous 2-ethyl
hexanoate in toluene solution. Preferably the molar percentages of
L-lactide, glycolide, and L(+)-tartaric acid is about 72/27/1,
respectively.
[0033] The L(+)-tartaric acid is previously dried, preferably over
silica gel in an Abderhalden drying apparatus for about 10 hours.
The reactor is then put under vacuum with stirring to remove
toluene. The reactor, under an atmosphere of oxygen-free nitrogen,
is then heated, preferably by immersing, it in an oil bath,
temperature=about 180.degree. C. to 190.degree. C., and stirring is
increased to about 125 rpm. Prior to immersion, a heating tape is
placed on the reactor lid. The time taken to completely melt the
reactor contents is noted, typically about 15 minutes for a load of
about 300 g at about 180.degree. C. Samples are taken every hour
during synthesis and analyzed by GPC to determine the percentage
residual monomer and to obtain values for average molecular weight
by number (Mn) and by weight (Mw) distributions. Typical reaction
times are of the order of about 9 to 15 hours. The final polymer is
also analyzed by titration to determine an acid number in meq/g and
by GC to determine residual unreacted monomer content. Further
analyses include IR (detection of characteristic C.dbd.O peak); NMR
(determination of lactide and glycolide content in polymer) and
residual tin (determination of residual tin due to use of stannous
2-ethyl hexanoate as catalyst).
Purification/Sodium salt formation of the above copolymer: Residual
monomer (typically <5% (W/W)) is removed and the copolymer is
converted to it's sodium salt form (to promote ionic salt
formation) in one step. The
poly-L-lactic-co-glycolic-co-L(+)-tartaric acid copolymer (PLGTA)
is dissolved in acetone by sonication in a sonication bath to give
a solution with a concentration in the range of 19-21% PLGTA by
weight.
[0034] To this solution is added a weak solution of an inorganic
base such as NaOH or Na.sub.2CO.sub.3, preferably 0.2M sodium
carbonate--Na.sub.2CO.sub.3 is used, in an amount so that the
resulting concentration of sodium is 1 to 2 times molar excess,
preferably 1.2 times molar excess, over copolymer carboxyl groups.
The solution is left to stir for about 15 to 60 minutes, preferably
30 minutes, at room temp. to aid sodium salt formation. It is then
fed at about 50 to 300 ml/min, preferably about 100 ml/min, into a
jacketed reactor containing de-ionized water cooled to about 1 to
4.degree. C., preferably 2.5.degree. C., using a circulation bath;
the amount of water is about 20 to 30 times volumetric excess over
acetone, preferably 20:1 volumetric excess over acetone. The water
is stirred at a rate sufficient to create surface turbulence in
order to avoid polymer agglomeration during precipitation using a
paddle linked to a stirrer motor.
[0035] Once precipitation is complete, the dispersion is left to
stir for a further 30 to 60 minutes to aid monomer removal before
being placed in centrifuge bottles and spun. The supernatant is
discarded and the cakes are resuspended in further de-ionized
water, re-spun and dried, preferably by lyophilization.
Preparation of a Compound (A) Polymer Ionic Conjugate: The
Synthesis Entails Binding Compound (A) to the copolymer sodium salt
in a medium in which both are soluble, preferably 3:1 (W/W)
acetonitrile:water, followed by precipitation of the resulting
ionic conjugate in de-ionized water and recovery of the
water-insoluble conjugate precipitate formed.
[0036] A solution of the acetate salt of Compound (A) in de-ionized
water is added to a solution consisting of a washed Na salt of
12,000 MW 71/28/1 to 73/26/1 PLGTA in acetonitrile (Range 24-26%
(W/W) solution) to which a weak base such as 0.5M Na.sub.2CO.sub.3
has been added so that it results in about a 1.05 molar excess of
Na over the acetate content of the Compound (A) acetate salt, and
left to stir for about 5 minutes to provide an alkaline
environment, preferably pH 8, to neutralize Compound (A)'s acetate
group. Approximate weight ratio of acetonitrile:water=3:1. Based on
target loading required (usually about 8% to about 12%), the
quantity of Compound (A) required is determined. From this the
volume of aqueous sodium carbonate required to neutralize the
acetate of Compound (A) is determined and finally the volume of
water for Compound (A) dissolution is calculated based on a desired
final acetonitrile:water (including sodium carbonate added)
volumetric ratio of about 3:1.
[0037] The Compound (A)-copolymer solution is left to stir for
about 10 to 15 mins. at about 0 to 5.degree. C., preferably
2.5.degree. C., to facilitate ionic binding and discourage covalent
binding (by use of low temperature) between the two components. The
solution is then fed at a rate of about 50 to 300 ml/min into about
a 20-30 to 1 volumetric excess of de-ionized water over the volume
of acetonitrile in the foregoing 3:1 acetonitrile-water solution,
stirred at a rate sufficient to provide surface agitation and avoid
agglomeration and cooled to about 1 to 4.degree. C., preferably
1.7.degree. C., in a jacketed reactor connected to a circulation
bath.
[0038] When precipitation is complete the dispersion is left to
stir for a further 30 to 60 minutes to aid removal of water-soluble
Compound (A)-oligomer compounds (oligomers are those lower
molecular weight fractions of PLGTA, which are undesirable since
they are water soluble) before being placed in centrifuge bottles
and spun at about 5000 rpm for about 15 minutes in a centrifuge.
The resultant centrifuge cakes are resuspended in de-ionized water
and re-spun. They are then frozen and dried by lyophilization for 2
days and Compound (I) (Compound (A) ionically bound to PLGTA) is
recovered. The loading is determined by HPLC analysis of the
supernatant for unbound Compound (A) and nitrogen analysis (the
Compound (A) nitrogen content is known and the polymer contains no
nitrogen whatsoever). Extraction of Compound (A) from Compound (I)
followed by HPLC analysis also allows determination of loading.
Compound (I) Nebulization: In order to provide a formulation
well-suited for injection into a patient, Compound (I) is
formulated into microspheres by dissolving it in ethyl acetate and
using ultrasonic atomization (a.k.a. nebulization) to spray the
solution into cold temperature, about -60.degree. C. to -78.degree.
C., ethanol, isopropanol or a mixture of hexane and isopropanol,
preferably isopropanol, which results in the formation of
microspheres of Compound (I) upon contact with the cold
isopropanol. The Compound (I) ethyl acetate solution can be
sterilized by passing it through a 0.2 .mu.m filter.
[0039] Compound (I) is dissolved in ethyl acetate preferably by
sonication/stirring to give about 8% to about 12% (W/W) solution,
preferably 12%, depending on polymer molecular weight and Compound
(A) loading, both of which may alter solution viscosity. This is
fed at about 4.90 ml/min. to 5.10 ml/min., preferably 5.00 ml/min.
to an industrial atomizer or nebulizer (Power--about 70%,
Amplitude--about 80%, Frequency--about 34 to 35 kHz, preferably
34.50 kHz; in general the nebulizer should be powerful enough to
generate a frequency which can uniformly spray (without "spitting")
the Compound (I) ethyl acetate solution from about 8% to about 12%
(W/W) in concentration, such concentrations lead to the formation
of solid microspheres and the frequency should be such that a mean
particle size of between 40 microns and 70 microns is obtained,
which will allow ease of injection through a 21-gauge or a 19-gauge
needle) and nebulized into a volume of isopropanol (IPA) that is 20
to 30 times, preferably 20 times, volumetric excess compared to the
ethyl acetate volume, cooled to about -60.degree. C. to about
-78.degree. C., cooling can be achieved, (e.g., via a reactor
jacket, addition of dry ice or insertion of a cooling coil) and
stirred at least at about 200 rpm (to avoid microsphere
agglomeration). De-ionised water at a temperature of about
6.degree. C. is fed at preferably 1.5 L/min to the nebulizer jacket
to eliminate any local heating effects which can cause fouling of
the nebulizer tip due to ethyl acetate evaporation. The solution
nebulized evenly and an off-white particulate dispersion is seen to
form in the IPA. This is allowed to thaw to about 0.degree. C. to
22.degree. C. over a period of about 30 mins. to 2 hrs before
passing it through a 125 .mu.m sieve (to remove any large
non-injectable droplets/particles) and on to a Whatman no. 1 filter
paper where it is vacuum-filtered. The filter cake is rinsed with
further IPA and then vacuum dried.
[0040] The present invention is illustrated by the following
example but is not limited by the details thereof.
EXAMPLE 1
Ionic Conjugate of P(I)LGTA (72/27/1) and Compound A
[0041] Step A: Synthesis of 300 g of P(I)LG/tartaric acid copolymer
(1-lactide:glycolide:tartaric acid=72:27:1)
[0042] A reactor was loaded with monomers of glycolide (Purac
Biochem, Netherlands, 68.71 g), lactide (Purac Biochem,
Netherlands, 227.53 g) and L(+)-Tartaric acid (Riedel-de Haen,
Seeize, Germany, article number 33801, 3.75 g) and stannous 2-ethyl
hexanoate (Sigma, St. Louis, Mo., USA, article number S-3252) in
toluene (Riedel-de Haen, Seelze, Germany) solution (0.0982M, 4.47
ml). This corresponded to molar percentages of 71.81%; 26.82%; and
1.36% respectively of L-lactide, glycolide, and L(+)-tartaric
acid.
[0043] The L(+)-tartaric acid was previously dried over silica gel
(Riedel-de Haen, Seelze, Germany) in an Abderhalden drying
apparatus for about 10 hours. The reactor (connected to a pump via
a liquid nitrogen trap) was then put under vacuum (0.04 mbar) with
stirring for about 50 minutes to remove toluene. The reactor, under
an atmosphere of oxygen-free nitrogen (BOC gases, Dublin, Ireland,
moisture content of 8 VPM), was then immersed in an oil bath
(Temperature=.about.180.degree. C.) and stirring was increased to
125 rpm. Prior to immersion, a heating tape (Thermolyne type 45500,
input control setting=4) was placed on the reactor lid. The time
taken to completely melt the reactor contents was noted, typically
about 15 minutes for a load of 300 g at about 180.degree. C.
Samples were taken every hour during synthesis and analyzed by GPC
to determine the percentage residual monomer and to obtain values
for average molecular weight by number (Mn) and by weight (Mw)
distributions. Typical reaction times were of the order of about 15
hours. The final polymer was also analyzed by titration to
determine an acid number in meq/g and by GC to determine residual
unreacted monomer content. Further analyses include IR (detection
of characteristic C.dbd.O peak); NMR (determination of lactide and
glycolide content in polymer) and residual tin (determination of
residual tin due to use of stannous 2-ethyl hexanoate as
catalyst).
Step B: Purification/Sodium Salt Formation with the above
Copolymer
[0044] Residual monomer (typically <5% (W/W)) was removed and
the copolymer was converted to it's sodium salt form (to promote
ionic salt formation) in one step. 81.05 g of a 12,000 g/mol
72/27/1 poly-L-lactic-co-glycolic-co-L(+)-tartaric acid copolymer
(acid number by titration=0.231 meq/g) was dissolved in 324.24 g of
acetone (Riedel-de Haen, Seelze, Germany) by sonication in a
sonication bath (Branson, Danbury, Conn., USA) to give a solution
with a concentration of 20.00% PLGTA by weight.
[0045] To this solution was added 56.17 ml of 0.2M Na.sub.2CO.sub.3
(Aldrich, Gillingham, Dorset, UK), thus providing a 1.2 times molar
excess of sodium over copolymer carboxyl groups. The solution was
left to stir for about 30 minutes at room temp. to aid sodium salt
formation. It was then fed at .about.100 ml/min into a 10 L
jacketed reactor containing 8.2 L of deionized water (approximately
a 20:1 volumetric excess over acetone cooled to about 2.5.degree.
C. using a circulation bath (Huber, Offenburg, Germany). This water
was stirred at 800 rpm to create surface turbulence and avoid
polymer agglomeration during precipitation using a paddle linked to
a stirrer motor.
[0046] Once precipitation was complete, the dispersion was left to
stir for a further 30 mins. to aid monomer removal before being
placed in centrifuge bottles and spun at 5000 rpm for about 15
minutes in a Sorvall centrifuge (DuPont Sorvall Products,
Wilmington, Del., USA). The supernatant was discarded and the cakes
were resuspended in further de-ionized water, respun and frozen in
a freezer (-13.degree. C.) overnight before being dried in a
small-scale lyophilizer (Edwards, Crawley, West Sussex, UK) the
next day. This lyophilizer contains no coolant system. After 5 days
of lyophilization 65.37 g of washed copolymer were recovered
representing a yield of 80.65%.
Step C: Preparation of Compound (I)
[0047] A solution of 1.27 g of the acetate, salt of Compound (A)
(Batch 97K-8501 from Kinerton Ltd., Dublin, Ireland, potency=85.8%
(potency refers to the percent free base peptide present in the
peptide acetate salt); acetate=10.87%) in 5.87 g of de-ionized
water was added to a solution consisting of 8.01 g of a washed Na
salt of 12,000 MW 72/27/1 PLGTA in 24.84 g acetonitrile (Riedel
de-Haen) (24.38% (W/W) solution to which 2.41 ml of 0.5M
Na.sub.2CO.sub.3 (this corresponds to a 1.05 excess of Na over the
acetate content of Compound (A)-acetate salt) had been added and
left to stir for about 5 minutes to provide an alkaline environment
(pH 8) for neutralization of Compound (A)'s acetate groups.
Approximate weight ratio of acetonitrile:water=3:1. Based on target
loading required, the quantity of Compound (A) required was
determined. From this the volume of aqueous sodium carbonate
required to neutralize the acetate of Compound (A) was determined
and finally the volume of water for Compound (A) dissolution was
calculated, based on a desired final acetonitrile:water (including
sodium carbonate added) volumetric ratio of 3:1.
[0048] The Compound (A)-copolymer solution was left to stir for
about 15 mins. at about 2.5.degree. C. to facilitate ionic and
discourage covalent binding between the two. The solution was then
fed at .about.100 ml/min into 630 ml (approximately a 20:1
volumetric excess over acetonitrile) of de-ionized water stirred at
350 rpm (to provide surface agitation and avoid Compound
(A)-copolymer agglomeration) and cooled to about 1.7.degree. C. in
a 6 L jacketed reactor connected to a circulation bath.
[0049] When precipitation was complete the dispersion was left to
stir for a further 30 minutes to aid removal of water-soluble
Compound (A)-oligomer compounds before being placed in centrifuge
bottles and spun at 5000 rpm for about 15 minutes in a Sorvall
centrifuge (DuPont Sorvall Products, Wilmington, Del., USA). The
resultant centrifuge cakes were resuspended in de-ionized water and
re-spun. They were then frozen and dried by lyophilization for 2
days. 8.30 g of the title product were recovered representing a
yield of 91.38%. The loading was determined by HPLC analysis of the
supernatant for unbound Compound (A) and nitrogen analysis (the
Compound (A) nitrogen content is known and the polymer contains no
nitrogen whatsoever). Extraction of Compound (A) from Compound (I)
followed by HPLC analysis also allows determination of loading,
which for this example was 11.25%.
Step D: Compound (I) Nebulization
[0050] 8.27 g of Compound (I) from step C was dissolved in 60.77 g
of ethyl acetate by sonication/stirring (room temp.) to give a
12.00% (W/W) solution. This was fed at 5 ml/min to an industrial
atomizer/nebulizer (Martin Walter Powersonic Model MW400GSIP,
available from Sodeva, France), Power=70%, Amplitude=80%,
Frequency=34.50 kHz and nebulized into 1.35 L of isopropyl alcohol
(IPA) (20 times volumetric excess compared to ethyl acetate volume)
cooled to about -74.+-.4.degree. C. (cooling achieved via reactor
jacket) and stirred at 200 rpm (to avoid microsphere agglomeration)
in a jacketed reactor. De-ionised water at a temperature of
6.degree. C. was fed at 1.5 L/min to the nebulizer jacket to
eliminate any local heating effects which can cause fouling of the
nebulizer tip due to ethyl acetate evaporation. The solution
nebulized evenly and an off-white particulate dispersion was seen
to form in the IPA. This was allowed to thaw to about 0.degree.
C.-4.degree. C. over a period of about 30 mins. to 2 hrs before
passing it through a 125 .mu.m sieve (to remove any large
non-injectable droplets/particles) and on to a Whatman no. 1 filter
paper where it was vacuum-filtered. The filter cake was rinsed with
further IPA and then vacuum dried. 6.88 g of injectable material
was obtained representing a yield of 83.19%. The microparticles of
Compound (I) had a mean particle size of about 54 microns.
[0051] The in vivo release of Compound (A) from microparticles of
Compound (I) can be and were tested according to the following
description. The in vivo study was designed to evaluate the in vivo
release profile of Compound (A) following the intramuscular
administration of microparticles of Compound (I) to male Beagle
dogs by means of the pharmacokinetic profile of Compound (A)
following its administration.
[0052] Pharmaceutical formulations of microparticles of Compound
(I) were administered intramuscularly in the rear legs muscles.
Following a single intramuscular administration of the irradiated
or non-irradiated prepared pharmaceutical forms containing
microparticles of Compound (I) (amount of microparticles injected
corresponded to 5 mg of Compound, (A) based upon the determination
that Compound (A) loading in Compound (I) was 11.23%) to groups of
six dogs each per pharmaceutical form. The quantitation of Compound
(A) in serum samples and the pharmacokinetic analysis is conducted
as follows.
[0053] Blood collection from the dogs are carried out before
injection (time 0), and 5, 15 and 30 min; 1, 2, 4, 8 and 12 hrs; 1,
2, 3 and 4-days; then twice per week over the first month (for
example on Mondays and Thursdays); and finally once a week from the
second month until completing the experiment, when the serum levels
of Compound (A) were no longer detected. However, the real times of
blood collection were recorded and used in the pharmacokinetic
analysis. Blood samples (5 ml) at times 0 (before i.m.
administration) and at 7, 21, 35, 56 and 84 days after the i.m.
injection and blood samples (4 ml) for the rest of sampling times,
were taken through the jugular or the cephalic veins at the
prescribed times.
[0054] The samples were placed in two fractions: one about 2.5 ml
or 3.5 ml in certain fixed time samplings, in tubes that contain
50, and 80 .mu.l, respectively, of a solution of aprotinin (10 ml
of Trasylol.RTM. 500000 KJU lypophillised and rediluted in 2 ml of
p.p.i. water) and the other one about 1.5 ml in tubes that were
allowed to stand.
[0055] After the red cells clot, the tubes were centrifuged for 20
min at 30000 r.p.m. at +4.degree. C. Serum with aprotinin were
removed and stored in two fractions at -20.degree. C. until the
sample was analyzed for Compound (A).
[0056] The concentration of Compound (A) in the serum samples were
analyzed by a radioimmunoassay method. Standard curves with blank
dog plasma and Compound (A) standard solutions were prepared daily.
In this method the limit of quantification for Compound, (A) in dog
serum samples is about 0.050 nanograms (ng)/ml. The areas under the
curve (AUC) and the maximum serum concentration (C.sub.max) were
normalized by the dose supplied (dose adminstered to each of the
animals expressed in .mu.g/kg). The index of the absorption rate
(C.sub.max/AUC) were also calculated.
[0057] The results of the foregoing experiment are shown in FIG.
1.
[0058] Compound (A) or a pharmaceutically-acceptable salt thereof,
Compound (I) or microparticles of Compound (I) can be administered
by oral, parenteral (e.g., intramuscular, intraperitoneal,
intravenous or subcutaneous injection, or implant), nasal, vaginal,
rectal, sublingual or topical routes of administration and can be
formulated with pharmaceutically acceptable carriers to provide
dosage forms appropriate for each route of administration.
[0059] Solid dosage forms, for oral administration include
capsules, tablets, pills, powders and granules. In such solid
dosage forms, the active compound is admixed with at least one
inert pharmaceutically acceptable carrier such as sucrose, lactose,
or starch. Such dosage forms can also comprise, as is normal
practice, additional substances other than such inert diluents,
e.g., lubricating agents such as magnesium stearate. In the case of
capsules, tablets and pills, the dosage forms may also comprise
buffering agents. Tablets and pills can additionally be prepared
with enteric coatings.
[0060] Liquid dosage forms for oral administration include
pharmaceutically acceptable emulsions, solutions, suspensions,
syrups, the elixirs containing inert diluents commonly used in the
art, such as water. Besides such inert diluents, compositions can
also include adjuvants, such as wetting agents, emulsifying and
suspending agents, and sweetening, flavoring and perfuming
agents.
[0061] Preparations for parenteral administration include sterile
aqueous or non-aqueous solutions, suspensions, or emulsions.
Examples of non-aqueous solvents or vehicles are propylene glycol,
polyethylene glycol, vegetable oils, such as olive oil and corn
oil, gelatin, and injectable organic esters such as ethyl oleate.
Such dosage forms may also contain adjuvants such as preserving,
wetting, emulsifying, and dispersing agents. They may be sterilized
by, for example, filtration through a bacteria-retaining filter, by
incorporating sterilizing agents into the compositions, by
irradiating the compositions, or by heating the compositions. They
can also be manufactured in the form of sterile solid compositions
which can be dissolved in sterile water, or some other sterile
injectable medium immediately before use.
[0062] Compositions for rectal or vaginal administration are
preferably suppositories which may contain, in addition to the
active substance, excipients such as coca butter or a suppository
wax.
[0063] Compositions for nasal or sublingual administration are also
prepared with standard excipients well known in the art.
[0064] It is preferred that the microparticles of Compound (I) be
administered via parenteral administration or oral
administration.
[0065] The effective dosage of the microparticles of Compound (I)
to be administered to a patient can be determined by the attending
physician or veterinarian and will be dependent upon the proper
dosages contemplated for Compound (A) and the loading of Compound
(A) in the microparticles of Compound (I). Such dosages will either
be known or can be determined by one of ordinary skill in the art.
Preferably the dosage should result in a level of at least 200
picograms/ml of Compound (A) in the patient.
[0066] The use of immediate or of sustained release compositions
depends on the type of indications aimed at. If the indication
consists of an acute or over-acute disorder, a treatment with an
immediate release form will be preferred over a prolonged release
composition. On the contrary, for preventive or long-term
treatments, a prolonged release composition will generally be
preferred.
[0067] Typically, the indication of upper gastrointestinal bleeding
will correspond an acute or over-acute treatment with a dosage of
about 80 to 120 .mu.g/day per person during approximately 5 days.
After endoscopical treatment, preventive treatment against
recurrence can be performed using microparticles of Compound (A) or
other sustained release forms as an adjuvant to usual
treatments.
[0068] For other indications other than upper gastrointestinal
bleeding, which require rather long term treatments, microparticles
of Compound (I) will be preferred.
* * * * *