U.S. patent number 3,911,098 [Application Number 05/441,695] was granted by the patent office on 1975-10-07 for medicament carrier.
This patent grant is currently assigned to American Cyanamid Company. Invention is credited to Richard Carl Capozza.
United States Patent |
3,911,098 |
Capozza |
October 7, 1975 |
Medicament carrier
Abstract
A controlled release insert for a living eye consisting of a
biologically effective form of pilocarpine and a biologically inert
biodegradable carrier consisting essentially of
poly[N-acetyl-6-O-(carboxymethyl)-D-glucosamine] gives effective
treatment to the human eye for prolonged periods. Other medicaments
and other enzymatically degradable forms of
poly(N-acetyl-D-glucosamine) may be used for rate controlled
release in the eye and other areas.
Inventors: |
Capozza; Richard Carl (Norwalk,
CT) |
Assignee: |
American Cyanamid Company
(Stamford, CT)
|
Family
ID: |
23753927 |
Appl.
No.: |
05/441,695 |
Filed: |
February 11, 1974 |
Current U.S.
Class: |
424/428 |
Current CPC
Class: |
A61K
47/36 (20130101); A61K 9/205 (20130101); A61K
9/0051 (20130101) |
Current International
Class: |
A61K
9/00 (20060101); A61K 9/20 (20060101); A61K
47/36 (20060101); A61K 009/52 () |
Field of
Search: |
;128/260,335.5
;424/19-22 |
References Cited
[Referenced By]
U.S. Patent Documents
Other References
Worden Rayon Textile Monthyl, Sept. 1941, XXII(9), p. 49,
"Developments in Organic Non-Cellulosic Fibrous Materials." .
Chem. Abstracts 79 No. 32258n, (1973). .
Chem. Abstracts 73 No. 43828a, (1970)..
|
Primary Examiner: Rose; Shep K.
Attorney, Agent or Firm: Walker; Samuel Branch
Claims
I claim:
1. A method of dispensing an eye drug over a prolonged period of
time comprising inserting in the conjunctival sac of the eye a
bioerodible enzymatically cleavable occular insert which is shaped
to conform to the curvature of the eye and is adapted for insertion
and retention in the conjunctival sac of the eye of an eye drug
intimately dispersed in an uncoated matrix directly contacting the
conjunctival sac of the eye of an enzymatically degradable form of
poly(N-acetyl-D-glucosamine) selected from the group consisting of
poly[N-acetyl-6-0-(carboxymethyl)-D-glucosamine],
poly[N-acetyl-6-0-(2'-hydroxyethyl)-D-glucosamine, and
poly[N-acetyl-6-0-(ethyl)-D-glucosamine], whereby the said form of
poly(N-acetyl-D-glucosamine) is slowly enzymatically degraded by
lysozyme in the tears over a period of time, and said eye drug is
slowly thereby released into tears, and contacts the eyeball.
2. The method of claim 1 in which the eye drug is pilocarpine free
base or a salt thereof.
3. An enzymatically degradable bioerodible eye drug delivery
occular insert device which is shaped to conform to the eye and is
adapted for insertion and retention in the conjunctival sac of the
eye for administering an eye drug to the eye of a living mammal
comprising: an uncoated matrix adapted to directly contact the
conjunctival sac of the eye consisting essentially of an
enzymatically degradable form of poly(N-acetyl-D-glucosamine)
selected from the group consisting of
poly[N-acetyl-6-0-(carboxymethyl)-D-glucosamine],
poly[N-acetyl-6-0-(2'-hydroxyethyl)-D-glucosamine], and
poly[N-acetyl-6-0-(ethyl)-D-glucosamine], and intimately dispersed
an at least slightly water soluble eye drug.
4. The occular insert of claim 3 in which the eye drug is
pilocarpine free base or a salt thereof.
5. A method for dispensing a free base form of an eye drug over a
prolonged period of time comprising inserting in the conjunctival
sac of the eye a bioerodible enzymatically cleavable occular insert
which is shaped to conform to the curvatuve of the eye and is
adapted for insertion and retention in the conjunctival sac of the
eye of the free base form of an eye drug intimately dispersed in
and ionically bound to an enzymatically degradable form of
poly[N-acetyl-6-0-(carboxymethyl)-D-glucosamine], whereby the said
poly[N-acetyl-6-0-carboxymethyl)-D-glucosamine] is slowly
enzymatically degraded over a period of time, and said free base
form of said eye drug is slowly thereby released into tears.
6. An enzymatically degradable bioerodible eye drug delivery
occular insert device which is shaped to conform to the eye and is
adapted for insertion and retention in the conjunctival sac of the
eye for administering the free base form of an eye drug to the eye
of a living mammal comprising: a matrix consisting essentially of
an enzymatically degradable form of
poly[N-acetyl-6-0-(carboxymethyl)-D-glucosamine], and intimately
dispersed therein and ionically bound thereto, the free base form
of an at least slightly water soluble eye drug.
Description
BACKGROUND OF THE INVENTION
This invention relates to the controlled release of drugs. The time
of release of medicaments or drugs can be in part controlled by
incorporating the drugs in a matrix of an enzymatically degradable
form of poly(N-acetyl-D-glucosamine) so that said form is slowly
enzymatically degraded over a period of time by body fluids and the
drug is released into the body fluids at the time of use for a
longer period than the drug would be released without the matrix
carrier.
The prior art shows many efforts over a long period by many
individuals to alter the rate of release of drugs. Where it is
considered that drugs may be administered to many areas and for
many different conditions and that various drugs have different
solubilities in both water and oil, and the period for desired
administration may vary from almost instantly to a long period of
up to and including years, it can be seen that there is a wide
range of medicaments and a wide range of conditions to be
controlled.
DESCRIPTION OF THE PRIOR ART
U.S. Pat. No. 2,552,027, Bird and Rochow, May 8, 1951, CASTING
Gelatine TABLETS, discloses the incorporation of medicaments,
particularly vitamins, into a gelatin-glycerine matrix, which are
"particularly useful for the administration of vitamins or other
pharmaceutical materials which should be released in the stomach or
digestive tract at comparatively slow rates."
In U.S. Pat. No. 3,604,417, Stolzenberg and Linkenheimer, Sept. 14,
1971, OSMOTIC FLUID RESERVOIR FOR OSMOTICALLY ACTIVATED LONG-TERM
CONTINUOUS INJECTOR DEVICE osmotic pressure is used to propel a
piston system so that a drug is slowly injected.
Many coating systems have been used on tablets for enteric release
in which various capsules have been coated either as a single
tablet or by coating particles in the tablets or capsules so that
drugs are released into the stomach or intestine at a controlled
rate and for a longer period than would result from the
administration of the medicament without such coatings.
U.S. Pat. No. 3,739,773, Schmitt and Polistina, June 19, 1973,
POLYGLYCOLIC ACID PROSTHETIC DEVICES, in Column 6, line 53, refers
to polyglycolic acid in combination with other products as slowly
digestible drug release devices. This patent in Column 7, lines 47
and following, mentions that dyes, antibiotics, antiseptics,
anesthetics, and other materials may be present in polyglycolic
acid devices. This last language also appears in Column 3, line 48
and following of U.S. Pat. No. 3,297,033, Schmitt and Polistina,
Jan. 10, 1967, SURGICAL SUTURES.
U.S. Pat. No. 3,435,008, Schmitt, Epstein and Polistina, Mar. 25,
1969, METHOD FOR PREPARATION OF ISOMERICALLY PURE .beta.-GLYCOLIDE
AND POLYMERIZATION METHOD FOR GLYCOLIDE COMPOSITIONS EMPLOYING
PARTIAL HYDROLYZATE OF SAID .beta.-GLYCOLIDE in Column 7, line 19
and following, discloses glycolide polymers as coatings for
medicaments to alter their digestive characteristics.
U.S. Ser. No. 179,129, filed Sept. 9, 1971, by Takeru Higuchi,
Anwar A. Hussain and John W. Shell, and the Netherlands rights to
which are assigned to the Alza Corporation, is referred to in the
Derwent Publications, LTD. Patent Index, and is available through
conventional documents in the file of Netherlands 7,212,272, in
Example 14 discloses a bioerodible ocular insert containing
pilocarpine free base using a matrix of polyglycolic acid.
Pilocarpine is mixed with polyvinyl alcohol and used as a core
between two sheets of polyglycolic acid. (see page 112). Page 60
discloses polyesters of lactic and glycolic acid as a carrier. Page
73, line 11, mentions "chitin" among other polysaccharides and
plant hydrocolloids. Presumably, the reference is to the naturally
occurring form of chitin. Claim 5 is drawn to bioerodibility by
enzymatic cleavage. Claim 14 is drawn to cross-linked gelatin.
Claim 50 is drawn to polylactic or polyglycolic release rate
controlling materials.
Sterile peanut oil and similar materials have been used as a
repository for penicillin for some time. The penicillin is slowly
released from the repository. Unfortunately, the peanut oil or
beeswax remains behind and is apt to form a sterile abscess rather
than be absorbed by tissues.
Carboxymethylchitin is disclosed in Carbohyd. Res. 7, 483-485
(1968), Ralph Trujillo.
This article mentions the hydrolysis of both chitin and
carboxymethylchitin by lysozyme.
Chitin has been estimated to be the second most abundant
polysaccharide in nature with a synthesis in the neighborhood of a
billion tons a year by marine organisms. See Chitin, N. V. Tracey,
Reviews of Pure and Applied Chemistry, Royal Australian Chemical
Institute, Vol. 7, No. 1, March 1957, pages 1 to 14.
The above patents and articles are herein hereby incorporated by
this reference thereto for background information on chitin, its
properties and derivatives.
Although it is well recognized that systems for the controlled
release of drugs are very much in demand, the wide range of
requirements is such that useful contributions are still being
sought and major efforts are being made by many research
organizations to improve drug delivery devices.
SUMMARY OF THE INVENTION
It has now been found that enzymatically degradable forms of
poly(N-acetyl-D-glucosamine) sometimes herein abbreviated as PAG,
are comparatively storage stable and resistant to hydrolytic
degradation so that medicaments may be incorporated and stored in a
matrix of such biodegradable form of PAG, and the medicament then
released in the tissue of living mammals by the enzymatic
degradation of the biodegradable form of PAG. The enzyme lysozyme
is particularly effective in the enzymatic degradation of the
biodegradable forms of PAG. Various forms of PAG may have different
degradation rates, and the degradation rate may vary with the
location of the drug release device.
Usually it is desired that the drug release device be mechanically
acceptable at a location of use. For instance, an ocular insert may
be designed to be placed adjacent to the eyeball inside the eyelid,
in the cul-de-sac of the conjunctiva between the schlera of the
eyeball and the lid. An insert needs to be soft so that it will
cause a minimum of irritation to the eyeball and the degradation
products are preferably such that they may be washed away by the
flow of tears without the necessity for removal of the device after
its drug content has been delivered. For other locations, such as
implantation beneath the surface of the skin or insertion in the
uterus as an anti-fertility device, the likelihood of irritation
from the mechanical aspects of the device are much less.
Because the requirements for use in the eye are among the more
rigorous, the present device will be described particularly in
conjunction with use in the eye although it is to be understood
that the device in its many forms may be used in other
locations.
N-acetyl-D-glucosamine has the formula: ##SPC1##
Groups below the plane of the paper are shown by a dotted bond.
Poly(N-acetyl-D-glucosamine) has ascribed to it the formula (ring
hydrogens omitted for clarity) ##SPC2##
Poly(N-acetyl-D-glucosamine) is a major component of naturally
occurring chitin. The naturally occurring material has not only the
poly(N-acetyl-D-glucosamine) but also inorganic salts thought to be
forms of calcium carbonate and proteinaceous material, the
composition of which is not presently known. The term "chitin" is
used herein to refer to the various naturally occurring forms of
chitin including the protein and inorganic carbonate components.
The term "purified chitin" is used to refer to chitin after
purification to remove calcium carbonate and other inorganic salts
and various proteins which may be present and is essentially
poly(N-acetyl-D-glucosamine). Some confusion exists in the
literature in that the name chitin is used as a name for
poly(N-acetyl-D-glucosamine) without specifying whether it is a
naturally occurring material containing inorganic salts and
proteins or whether the term is intended to designate purified
poly(N-acetyl-D-glucosamine) without specifying the degree of
purity or the character of the impurities present.
The term "enzymatically degradable form of
poly(N-acetyl-D-glucosamine)" refers both to the purified
poly(N-acetyl-D-glucosamine) from chitin itself as well as the
carboxymethyl, hydroxyethyl, and 0-ethyl derivatives, etc.
The carboxymethyl derivative, properly called
poly[N-acetyl-6-0-(carboxymethyl)-D-glucosamine] has the formula
##SPC3##
The hydroxyethyl derivative, properly called
poly[N-acetyl-6-0-(2'-hydroxyethyl)-D-glucosamine] has the formula
##SPC4##
The 0-ethyl derivative, properly called
poly-[N-acetyl-6-0-(ethyl)-D-glucosamine] has the formula
##SPC5##
The above forms are sometimes hereinafter designated by the Roman
Numeral below the formula.
Other similar derivatives which are enzymatically degradable,
particularly by lysozyme, are included within the generic term
"enzymatically degradable form of
poly(N-acetyl-D-glucosamine)."
Because of the nature of the polymers, carboxymethylation,
hydroxyethylation, or ethylation may not be 100%, and may in part
occur on the 3-hydroxyl. Unless otherwise specified, under or
over-substitution of the
poly[N-acetyl-6-0-(carboxymethyl)-D-glucosamine] is to be included
as a biodegradable form of PAG. The solubility in a specified
solvent is one test of the degree of substitution. For example, the
0-ethyl derivative is water soluble when the ethyl group to
glucosamine ratio is about 1 and organic soluble when the degree of
substitution is greater than 1.
The term "drug" is used to refer to a substance other than a food
intended to affect the structure or function of the body of man or
other animal. The term is somewhat broader than "medicine" in that
the term "medicine" is sometimes considered to be restricted to an
agent which is administered to affect or control a pathogenic
condition. The broader term "drug" here is also used to include
steroids and other fertility controlling agents which may be
incorporated in an intrauterine contraceptive device or other
materials which may be included to affect the fertility of females
or males either as an intrauterine device or subcutaneously.
The term "dispensing" is used to designate a method of
administering a drug to man or other animal and includes the
release of the drug to a desired location. This would include the
eye, gastrointestinal tract (alimentary), intrauterinely,
intramuscularly, subcutaneously, or into the mucosa of the nose,
mouth (sublingual), or rectum, etc. The release over a prolonged
period of time designates any decrease in the release rate of the
drug over that which would be expected if the drug were
administered alone and would include from the matter of a few
minutes as, for example, in an ocular insert containing pilocarpine
to a duration of six months to a year which might be desired for
the administration of a steroid in an intrauterine contraceptive
device. For some conditions, even a longer period of
administration, such as the lifetime of the patient, could be
desired but usually a period of a very few hours up to about six
months includes the medically preferred range.
Because the enzymatically degradable form of
poly-(N-acetyl-D-glucosamine) is a solid which can be removed, a
long-acting repository pellet for insertion beneath the skin is
quite practical as if for any medical reason it is desired to
discontinue administration of the drug, the insert with the
remaining drug charge may be removed simply by excision.
The term "enzymatically degradable" refers to a form of
poly(N-acetyl-D-glucosamine) or its derivatives which is broken
down into body fluid soluble components and which are washed out as
in tears, or transported elsewhere by tears, or other body fluid,
and later degraded further or metabolized by the body or excreted
by the body. The problem of retention by the body or disposal of
the residual matrix is minimal or non-existent.
While other enzymes may also affect the enzymatic degradation of
the poly(N-acetyl-D-glucosamine) matrix, the enzyme which is most
widely distributed in the body and here very effective is lysozyme.
Lysozyme occurs in practically all of the body fluids, particularly
the tears, and effectively breaks down the polymer chain to water
soluble or disposable components.
Chitosan, which is a common name for the deacylated form of
poly(N-acetyl-D-glucosamine), and which is poly(D-glucosamine) is
not enzymatically degradable by lysozyme.
By contrast, the present enzymatically degradable forms of
poly(N-acetyl-D-glucosamine) are not readily hydrolyzed by water.
For instance, I in a phosphate buffer at pH 7.2 at 37.degree.C for
24 hours is not hydrolyzed whereas under the same time and
temperature in the presence of lysozyme hydrolysis occurs.
It is highly advantageous to have the degradation of the
enzymatically degradable form of poly(N-acetyl-D-glucosamine) occur
only by the action of an enzyme as the resistance to hydrolytic
degradation markedly reduces problems of manufacture and storage in
the presence of ambient moisture, and ensures a steady smooth
surface erosion rather than a fragmentation process commonly
experienced by polymers which are hydrolyzed by small
molecules.
Any of the drugs used to treat the eye and surrounding tissues can
be incorporated with the enzymatically degradable form of PAG of
this invention. Also, it is practical to use the eye and
surrounding tissues as a point of entry for systemic drugs that
enter circulation in the blood stream and produce a pharmacological
response at a site remote from the point of application of drug and
the enzymatically degradable form of PAG matrix. Thus, drugs which
will pass through the eye or the tissue surrounding the eye to the
bloodstream, but which are not used in therapy of the eye itself,
can be incorporated in the enzymatically degradable PAG matrix.
Suitable drugs for use in therapy of the eye with the present
insert include, without limitation: Anti-infectives: such as
antibiotics, including tetracycline, chlortetracycline, bacitracin,
neomycin, polymyxin, gramicidin, oxytetracycline, chloramphenicol,
and erythromycin; sulfonamides, including sulfacetamide,
sulfamethazole, and sulfisoxazole; antivirals, including
idoxuridine; and other anti-infectives including nitrofurazone and
sodium propionate; Antiallergenics such as antazoline,
methapyrilene, chlorpheniramine, pyrilamine and prophenpyridamine;
Anti-inflammatories such as hydrocortisone, hydrocortisone acetate,
dexamethasone, triamcinolone, medrysone, prednisolone, prednisolone
21-phosphate and prednisolone acetate. Decongestants such as
phenylephrine, naphazoline, and tetrahydrazoline; Miotics and
anticholinesterases such as pilocarpine, eserine salicylate,
carbachol, disopropyl fluorophosphate, phospholine iodide, and
demecarium bromide; matropine, scopolamine, tropicamide,
eucatropine, and hydroxyamphetamine and sympathominetics such as
epinephrine. Drugs can be in various forms, such as unchanged
molecules, components of molecular complexes, or nonirritating,
pharmacologically acceptable salts, such as hydrochloride,
hydrobromide, sulfate, phosphate, nitrate, borate, acetate,
maleate, tartrate, salicylate, etc. Furthermore, simple derivatives
of the drugs (such as ethers, esters, amides, etc.) which have
desirable retention and release characterics but which are easily
hydrolyzed by body pH, enzymes, etc. can be employed. The amount of
drug incorporated in the ocular insert varies widely, depending on
the particular drug, the desired therapeutic effect, and the time
span for which the ocular insert will be used. Since the ocular
insert is intended to provide the complete dosage regime for eye
therapy for but a particular time span, such as 24 hours, there is
no critical upper limit on the amount of drug incorporated in the
device. The lower limit will depend on the activity of the drug and
its capability of being released from the device. Thus, it is not
practical to define a range for the therapeutically effective
amount of drug incorporated into the device. However, typically,
from 1 microgram to 1 milligram of drug is incorporated in each
insert.
In each case, the polymeric material used to form the ocular insert
is chosen for its compatibility with a particular drug and its
capability of releasing that drug at an appropriate rate over a
prolonged period of time. Specific, but nonlimiting, examples of
combinations of drugs and polymers for use in forming the ocular
insert include: poly-[N-acetyl-6-0-(carboxymethyl)-D-glucosamine]
and epinephrine; poly[N-acetyl-6-0-(carboxymethyl)-D-glucosamine]
and mixture of pilocarpine hydrochloride and epiniphrine;
poly[N-acetyl-6-0-(2'-hydroxyethyl)-D-glucosamine] and
acetazolamide; poly[N-acetyl-6-0-(ethyl)-D-glucosamine] and
phospholine iodide;
poly[N-acetyl-6-0-(carboxymethyl)-D-glucosamine] and triamcinolone,
or in general any of the drugs listed above and the enzymatically
degradable form of poly(N-acetyl-D-glucosamine), including degrees
of substitution greater or less than 1, and related derivatives
such as other lower alkyl derivatives instead of
poly[N-acetyl-6-0-(ethyl)-D-glyucosamine], other carboxyalkyl
derivatives, and their esters and salts, hydroxyalkyl derivatives,
etc.
The degradation rate of the enzymatically degradable form of
poly(N-acetyl-D-glucosamine) can be lowered by cross-linking, if a
slower release rate is preferred.
The ocular insert can be fabricated in any convenient shape for
comfortable retention in the cul-de-sac. It is important, however,
that the device have no sharp, jagged, or rough edges which can
irritate the sensitive tissues of the eye. Thus, the marginal
outline of the ocular insert can be ellipsoidal, bean-shaped,
rectangular, etc. In cross section, it can be concavo-convex,
rectangular, etc. As the ocular insert is flexible and, in use,
will assume essentially the configuration of the scleral curvature,
the original shape of the device is not of controlling importance.
Dimensions of the device can vary widely. The lower limit on the
size of the device is governed by the amount of the particular drug
to be applied to the eye and surrounding tissues to elicit the
desired pharmacological response, as well as by the smallest sized
device which conveniently can be inserted and removed from the eye.
The upper limit on the size of the device is governed by the
limited space within the cul-de-sac that conveniently and
comfortably can be filled with an ocular insert. Typically, the
ocular insert is 4 to 20 millimeters in length, 1 to 12 millimeters
in width, and 0.1 to 1 millimeter in thickness. Preferably, it is
ellipsoidal in shape and about 6 .times. 4 .times. 0.5 millimeters
in size.
While particularly convenient for an insert in the eye, the matrix
containing the drug of the present invention can include other
drugs for other areas. For instance, if the drug is to be taken
orally, a tablet of a size and shape adapted to being swallowed is
preferred. If it is to be placed subcutaneously, a tablet or rod
such that it may be placed under the skin in an appropriate
location is selected. The amount of drug and the time over which it
is to be dispensed are controlling in the choice of size of the
implant.
While the drug may be combined with the enzymatically degradable
form of PAG matrix in any convenient way, it is particularly
convenient to dissolve both in a common solvent which permits
casting of the enzymatically degradable form of PAG as a matrix
containing the drug to be dispersed therein.
Poly(N-acetyl-D-glucosamine) is reported to be insoluble in all
solvents except 88% phosphoric acid which badly degrades the
polymer. Unexpectedly, it has now been found that
hexafluoroisopropanol (HIPA) and hexafluoroacetone sesquihydrate
(HFAS) are solvents for the polymer. These are extremely powerful
solvents, and so much so that care must be used in selecting drugs
which are compatible with such solvents to form solutions for
casting.
Poly[N-acetyl-6-0-(carboxymethyl)-D-glucosamine], I,
poly[N-acetyl-6-0-(2'-hydroxyethyl)-D-glucosamine], II, and
poly[N-acetyl-6-0-(ethyl)-D-glucosamine], III, are preferred
because of cosolubility with many drugs in common solvents,
including water. Non-toxic solvents are preferred.
I and II are water soluble at the 5% level, and III is water
soluble at the 5% level if the degree of substitution is not more
than about 1, and organic solvent soluble if more than about 1.
Organic solvents may be used such as alcohols, chloroform, benzene,
toluene, mixtures of benzene and toluene with alcohols and
ketones.
Pilocarpine or other drugs can be incorporated into matrices of
these enzymatically degradable forms of PAG by hydrogen bonding,
covalent bonding, ionic bonding or simple entrapment. The matrices
themselves can be variably crosslinked with a variety of physical
and chemical agents. They can be sterilized and when hydrated
become quite pliable, while retaining adequate strength to resist
manipulation.
Present day therapy of topical drug application consists of drops
and ointments. There are several deficiencies associated with these
methods of delivery -- (1) it is impossible to achieve 24 hour
control of the disease (2) it is wasteful with respect to the
amount of drug used (3) some people show strong sensitivity to
cholinergic and adrenergic drops (4) many patients fail to apply
the medication as directed resulting in poor control of the disease
(5) side effects result from the drug passing through the lachrymal
duct into the circulatory system. The herein described invention
eleminates these problems and provides a means of releasing
medication into the tear films in therapeutic levels continuously.
The device is biodegradable and, hence, it is not necessary to
remove from the eye and also capable of delivering large dosages
giving it broad drug applicability. This invention constitutes a
more efficient means of drug delivery that prolongs and enhances
the drug effect.
As the scope of this invention is broad, it is illustrated by the
following typical examples in which temperatures are centigrade,
and parts are by weight unless clearly otherwise specified.
EXAMPLE 1
Purification of Chitin
A commercial grade of chitin (Cal-Biochemicals) was finely ground
in a ball mill overnight to pass a 6 mm screen and be retained by a
1 mm screen. 149 g. of this finely ground material was decalcified
by extracting with 825 ml. of 2N HCl at 4.degree.C for 48 hours, in
a flask stirred with a magnetic stirrer. The material was collected
by centrifugation and washed repeatedly with water until neutral.
The ash content was 0.4-0.5%. The decalcified chitin was then
stirred at room temperature with 1500 ml. of 90% formic acid
overnight. The mixture was centrifuged and the residue repeatedly
washed with water. The washed chitin was then suspended in 2 l. of
10% NaOH solution and heated at 90.degree.-100.degree.C. for 2.5
hours. The solution was filtered, the cake washed with water until
neutral, washed several times with absolute ethanol and ether, and
dried at 40.degree.C. under reduced pressure; yield 66 g. of
poly(N-acetyl-D-glucosamine). Infrared spectrum (KBr pellet) shows
bands at 3500 cm.sup..sup.-1 (S), 2900 (W), 1652 (S), 1619 (S),
1550 (S), 1370 (S), 1300 (M), 1070 (Broad). (S is strong, M is
medium, W is weak).
EXAMPLE II
Poly[N-acetyl-6-0-(carboxymethyl)-D-glucosamine]
15 g. of the poly(N-acetyl-D-glucosamine) from Example 1 was
swollen with 100 ml. of dimethylsulfoxide (DMSO). To this highly
swollen suspension was added 400 ml. of 2-propanol and the mixture
was stirred vigorously under nitrogen while 40 ml. of 30% aqueous
NaOH was added over an interval of 30 minutes at room temperature.
After stirring for an additional hour, 18 g. of chloracetic acid
dissolved in 40 ml. of water was added dropwise over a 30 minute
period. The mixture was then heated at 55.degree.C. for 24 hours.
The mixture was decanted and to the residue was added 100 ml. of
70% methanol. The suspension was then neutralized with 5 ml. of 90%
acetic acid. The mixture was filtered, washed with 70% methanol,
absolute methanol and dried at 40.degree.C. in vacuo. Yield 24 g.
of poly[N-acetyl-6-0-(carboxymethyl)-D-glucosamine], I. Infrared
(KBr pellet) shows bands at 3500 cm.sup..sup.-1. (S), 2900 (M),
1600 Broad (S), 1400 (M), 1320 (M), 1100 Broad (S). A sample was
titrated and shown to have 4.03 meq acid/g indicating 100% of the
repeating mers were carboxylated. Films easily removed from glass
were cast from water solution and shown to be transparent, flexible
and tough.
EXAMPLE III
Preparation of poly(D-glucosamine)
A procedure similar to that described by P. Broussignoc, Chemie and
Industrie, 99 (9) (68), 1243 was used. To a solution of 180 g. of
96% ethanol and 180 g. ethylene glycol was added 360 g. KOH with
stirring. To this solution was then added 54 g. of
poly(N-acetyl-D-glucosamine) (purified Chitin) from Example I and
the mixture heated at 120.degree.C. for 6 hours. After cooling an
equal volume of water was added to the mixture. The mixture was
filtered and washed several times with water until neutral, then
twice with acetone, and dried in vacuo. Yield 42.6 g. of
poly(D-glucosamine), sometimes called chitosan. Infrared spectrum
(KBr pellet) showed bands at 3450 cm.sup..sup.-1. (S), 2900 (M),
1620 (S), 1600 (S), 1370 [Broad (S)], 1050 [Broad (S)]. Upon
potentiometric titration of the sample 81.4% of the mers were found
to be deacylated. The product is soluble in 3% acetic acid and
forms clear, flexible, tough films from this solution. It is not
enzymatically biodegradable by lysozyme.
EXAMPLE IV
Poly(D-Glucosamine)/Pilocarpine Film
To 5 ml. of 3% acetic acid was added 0.25 g. of poly(D-glucosamine)
from Example III. To the solution thus formed was then added 50 mg.
pilocarpine free base and 100 ul of tritiated pilocarpine, and the
mixture was cast as a film (40 mil wet thickness) on glass. This
film was crosslinked by dipping the film in 37% formaldehyde
solution for 5 hours. This film showed zero order release over a
period of 3 days at which time it was still releasing pilocarpine
at a zero order rate. About 70 percent of the pilocarpine remained
in the film matrix after 3 days. The use of tritiated pilocarpine
permits the use of a liquid scintillation counter to monitor the
release rate accurately and conveniently. Radiological hazards are
associated with such tritiated material in the treatment of human
subjects so experimental animals are preferred to study release
rates.
EXAMPLE V
Poly[N-Acetyl-6-0-(Carboxymethyl)-D-Glucosamine]/Pilocarpine
Film
To a 5% solution of poly[N-acetyl-6-0-carboxymethyl)-D-glucosamine]
(0.95 g.) in water was added 50 mg. of pilocarpine nitrate and 100
ul of tritiated pilocarpine. A film 40 mils thick was cast on a
glass plate and allowed to dry. The film was crosslinked by dipping
into 10% alum for 5 hours. Release of pilocarpine from this film in
an aqueous solution approximating human tears is essentially first
order, with 90% of the pilocarpine being released within about 5
hours.
EXAMPLE VI
Poly(N-Acetyl-D-Glucosamine) Matrix
Membranes of poly(N-acetyl-D-glucosamine) were prepared by
dissolving poly(N-acetyl-D-glucosamine) in each of
hexafluoroacetone sesquihydrate (1.4% solution) and
hexafluoroisopropanol (2% solution). The films were tough,
transparent, non-tacky, flexible and were quite pliable when
hydrated yet retained adequate strength to resist manipulation. The
membranes showed no hydrolysis after exposure to water for 5 days.
In the presence of lysozyme, however, the films were degraded
slowly. The films eroded release any drug in the film slowly.
EXAMPLE VII
Biodegradability of
Poly[N-Acetyl-6-0-(Carboxymethyl)-D-Glucosamine]
After 24 hours incubation at 37.degree.C. in phosphate buffer pH
7.2 containing 1500 units/ml of lysozyme,
poly[N-acetyl-6-0-(carboxymethyl)-D-glucosamine] was hydrolyzed to
oligomers as determined by Gel Permeation Chromatography. A control
containing no enzyme was not hydrolyzed under the same
conditions.
EXAMPLE VIII
In Vivo Results Using
Poly[N-Acetyl-6-0-(Carboxymethyl)-D-Glucosamine]/Pilocarpine
Membranes of poly[N-acetyl-6-0-(carboxymethyl)-D-glucosamine] were
evaluated in vivo for sustained pharmacological effect and eye
irritation. In the right eye of each of three rabbits was placed a
1 mm .times. 10 mm film strip of
poly[N-acetyl-6-0-(carboxymethyl)-D-glucosamine] (0.25 g.
poly[N-acetyl-6-0-(carboxymethyl)-D-glucosamine]/0.64 g.
pilocarpine). Within 15 minutes after implantation of the film
strip a substantial lowering in pupillary constriction was observed
and lasted approximately 6 hours. The membranes were well tolerated
and slowly eroded in the eye. Such a prolonged effect in rabbits
can be extrapolated to a 24 hour effect in humans since the rabbit
metabolizes pilocarpine more rapidly than a human.
EXAMPLE IX
Poly[N-Acetyl-6-0-(2'-Hydroxyethyl)-D-Glucosamine]
Into a screw cap bottle was placed 13.6 g. of purified PAG milled
so that it passes a 1 mm. sieve. To the bottle was added 200 ml. of
cold (0.degree.-5.degree.C.) aqueous 43% NaOH and the contents
stirred for 2 hours under nitrogen and then held at
0.degree.-4.degree.C. for 10 hours. The swollen alkali derivative
was then squeezed to 3 times its original weight in a sintered
glass funnel, disintegrated and frozen at -20.degree.C. under
nitrogen for 1 hour and then thawed at room temperature for 1 hour.
The freeze-thaw cycle was repeated 3 times. To the alkali
derivative was then added 120 ml. of dimethylsulfoxide (DMSO) and
the slurry added immediately to a stirred autoclave. The autoclave
was purged several times with nitrogen and 53.2 ml. of ethylene
oxide was added (16 equivalents/equivalent of PAG). The mixture was
held at 50.degree.C. for 18 hours. The solution was then carefully
neutralized with glacial acetic acid, dialyzed and then
lyophilized.
The hydroxyethyl derivative can be further purified by
precipitating the polymer from aqueous solution with acetone. A
freshly precipitated sample of
poly[N-acetyl-6-0-(2'-hydroxyethyl)-D-glucosamine] readily
dissolved in water, 5% aqueous sodium hydroxide, and 3% acetic acid
and is precipitated from these solutions by acetone. Samples
analyzed for C, H and N showed the composition to be one in which
1.5 hydroxyethyl groups had been substituted per glucosamine
residue.
EXAMPLE X
Poly[N-Acetyl-6-0-(Ethyl)-D-Glucosamine]
The procedure of Example IX was followed except 75 ml. of
ethylchloride was added instead of ethylene oxide and the reaction
held at 50.degree.C. for 15 hours. A water soluble derivative is
obtained.
To obtain an organic soluble derivative, the ethylchloride was
mixed with benzene (75% of the amount of ethylchloride). The
reaction time was 10 hours and the temperature was controlled as
follows: 1 hour heating up to 60.degree.C., 1 hour heating up to
80.degree.C., 1 hour heating up to 130.degree.C. and 7 hours at
130.degree.C. An organic solvent soluble product was obtained. The
following solvents are useful for solubilization (5% solution) of
this polymer at room temperature: 0-xylene, benzene, toluene,
methylethyl ketone, 1.4 mixture of alcohol and benzene, chloroform
and alcohols.
In the following example using pilocarpine free base, the drug is
bound ionically to the polymer. The attractive features of such a
system are (1) slower drug delivery and (2) capability of
delivering pilocarpine as a free base which, as such, has a higher
potency. Up to now, it was not possible to deliver pilocarpine as
the free base since it is unstable in this form and as a result is
usually delivered as the hydrochloride or nitrate salt.
EXAMPLE XI
Pilocarpine/Poly[N-Acetyl-6-0-(Carboxymethyl)-D-Glucosamine]
Inserts
A 5% solution of poly[N-acetyl-6-0-(carboxymethyl)-D-glucosamine]
was prepared in deionized water. The solution was acidified with
acetic acid and the polymer precipitated by slowly adding this
solution to acetone. The polymer was dried in vacuo at 40.degree.C.
overnight. Films were prepared from 5% aqueous solutions containing
the following relative weights:
Poly[N-Acetyl-6-0-(Carboxy- Drug Dose per Pilocarpine
methyl)-D-Glucosamine 1.5 mg. Strip
______________________________________ 9.1 mg 90.9 mg 0.10 mg 19.4
mg 80.6 mg 0.24 mg 33.3 mg 66.6 mg 0.50 mg
______________________________________
The films were cut into strips approximately 1 mm .times. 10 mm
weighing 1.5 mg each. In this manner, the drug dosages are
delivered from each respective strip, when inserted in the eye.
Effective medication for a treatment day is obtained by placing an
insert 1 mm by 10 mm in the human eye.
* * * * *