U.S. patent number 3,911,116 [Application Number 05/380,711] was granted by the patent office on 1975-10-07 for process for promoting wound healing with chitin derivatives.
Invention is credited to Leslie L. Balassa.
United States Patent |
3,911,116 |
Balassa |
* October 7, 1975 |
Process for promoting wound healing with chitin derivatives
Abstract
Wound healing compositions and the process of healing wounds
with such compositions are described, the compositions containing a
chitin derivative.
Inventors: |
Balassa; Leslie L. (Blooming
Grove, NY) |
[*] Notice: |
The portion of the term of this patent
subsequent to January 4, 1989 has been disclaimed. |
Family
ID: |
26703102 |
Appl.
No.: |
05/380,711 |
Filed: |
July 19, 1973 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
Issue Date |
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27977 |
Apr 13, 1970 |
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619007 |
Feb 27, 1967 |
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704538 |
Feb 12, 1968 |
3632754 |
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Current U.S.
Class: |
514/55; 424/538;
424/447 |
Current CPC
Class: |
A61K
31/715 (20130101); C08B 37/003 (20130101) |
Current International
Class: |
A61K
31/715 (20060101); C08B 37/00 (20060101); C08B
37/08 (20060101); A61K 031/70 () |
Field of
Search: |
;424/180,28,95 |
References Cited
[Referenced By]
U.S. Patent Documents
Primary Examiner: Friedman; Stanley J.
Assistant Examiner: Stephens; Daren M.
Attorney, Agent or Firm: Darby & Darby
Parent Case Text
This is a continuation of application Ser. No. 27,977, filed Apr.
13, 1970 and now abandoned, which in turn is a continuation-in-part
of my copending application Ser. No. 619,007 filed Feb. 27, 1967,
now abandoned, and a continuation of application Ser. No. 704,538
filed Feb. 12, 1968, now U.S. Pat. No. 3,632,754.
Claims
What is claimed is:
1. A process for facilitating healing of a wound in a mammal which
comprises topically administering a therapeutically effective
dosage of a material selected from the group consisting of hydroxy
lower alkyl chitin, carboxy lower alkyl chitin, lower alkyl chitin,
chitin acetate, chitin phosphate, chitin nitrate, a salt of carboxy
lower alkyl chitin and chitin citrate.
2. A process for accelerating the healing of a surgical wound in a
mammal requiring healing which comprises administering to said
wound a therapeutically effective dosage of a finely divided
material selected from the group consisting of hydroxy lower alkyl
chitin, carboxy lower alkyl chitin, lower alkyl chitin, chitin
acetate, chitin phosphate, chitin nitrate, a salt of carboxy lower
alkyl chitin and chitin citrate.
3. A process for facilitating healing of a wound in a mammal which
comprises parenterally administering a therapeutically effective
dosage of a material selected from the group consisting of hydroxy
lower alkyl chitin, carboxy lower alkyl chitin, lower alkyl chitin,
chitin acetate, chitin phosphate, chitin nitrate, a salt of carboxy
lower alkyl chitin and chitin citrate.
4. A process according to claim 2 which comprises topically
administering said finely divided material at the site of said
wound.
Description
This invention relates to methods of promoting the healing of
wounds and compositions therefor comprising chitin derivatives.
Medicine has long been interested in improving the healing of
wounds. Patients suffering from diabetes or undergoing extensive
cortisone treatment show extremely slow rates of healing of any
wounds which they receive. Thus, surgery on such patients involves
additional risks not present with other patients. Moreover, rapid
healing of wounds is particularly desired for patients in tropical
countries where the risk of infection is high. Rapid healing is
also desired in the case of soldiers who have been wounded in a
battle zone and cannot easily and quickly be removed therefrom.
Acceleration of wound healing is highly desirable in the case of
patients who cannot readily be immobilized, such as farm
animals.
In evaluating the utility of a material to promote wound healing, a
reproducible test is necessary to give comparative data. Such a
test method has been described by Prudden et al in: "The
Acceleration of Wound Healing with Cartilage," Surgery, Gynecology
and Obstetrics, 105:283 (1957). In this method, rats are tested in
pairs, each pair receiving an identical surgical incision, only the
one rat of the pair receiving a measured dose of the material whose
wound healing properties is to be determined. The pair is then kept
in the same cage and the tensile strength of the wounds in the two
rats is determined in millimeters of mercury. The difference in the
tensile strengths between the treated rat and the control rat is
expressed as the percentage improvement obtained. Considering
biological variance it is believed that only differences of about
10% or more are significant.
There have been several recent developments reported concerning
materials which promote wound healing. In this connection U.S. Pat.
No. 3,232,836 describes the parenteral administration of
N-acetylglucosamine as a wound healing material. Utilizing the test
method of Prudden et al referred to in the preceding paragraph,
N-acetylglucosamine showed improvement in tensile strength of only
about 10% whereas Prudden and his co-workers have reported
significantly larger increases in wound healing by the use of
cartilage preparations from various animals. Depending on the age
and species of animal and the fineness of the cartilage powder,
improvements ranging from 20 to 40% in wound healing tensile
strength have been reported by Prudden.
Now it has been discovered that finely divided chitin, partially
depolymerized chitin, and chitin derivatives possess the ability to
promote the healing of wounds.
Accordingly, one aspect of the present invention relates to novel
methods of promoting and assisting the healing of wounds as, for
example, damaged mammalian tissue, open ulcers, etc., and to
compositions therefor.
Another aspect of the invention relates to significant improvements
in wound healing strength achieved by the administration of finely
divided chitin, partially depolymerized chitin or chitin
derivatives to a patient.
An additional aspect of the present invention is concerned with
articles of manufacture such as surgical bandages, surgical
sutures, etc., containing the wound healing materials of the
present invention.
These and other aspects of the present invention will be apparent
from the following description.
Chitin is a polysaccharide, believed to be poly
(N-acetylglucosamine) which forms the cell walls of fungi and the
hard shell of insects and crustaceans. As used herein, the term
"chitin" embraces naturally occurring chitin, synthetic chitin, as
well as poly (N-acetylglucosamine) and its epimer poly
(N-acetylgalactosamine). The partially depolymerized chitin, e.g.
chitotriose, chitobiose, is a substance which retains its polymeric
nature but has undergone a reduction in molecular weight (i.e.,
chain length) as a result of (1) enzymatic action such as by a
chitinase enzyme, (2) chemical treatment such as acid hydrolysis or
alkaline treatment, and (3) physical treatment.
The chitin derivatives contemplated are materials such as ethers
formed with pharmaceutically acceptable radicals and esters or
salts with pharmaceutically acceptable acids. Examples of suitable
derivatives include hydroxy lower alkyl chitin such as hydroxyethyl
chitin, carboxy alkyl chitin such as carboxymethyl chitin, salts of
carboxy lower alkyl chitin such as the zinc salt, lower alkyl
chitin such as methyl chitin and ethyl chitin, chitin acetate,
chitin nitrate, chitin citrate, chitin phosphate, N-acyl
derivatives derived from monocarboxylic aliphatic acids such as
N-formyl, N-acetyl, N-propionyl, N-caproyl, etc.
It is preferred to use natural chitin as the wound healing
accelerator. The naturally occurring chitin is preferably chitin of
fungal origin, both by reason of its ready availability and its
high degree of effectiveness.
The degree of improvement in wound healing obtained with the chitin
materials is at least equal to and in many instances greater than
that derived from the cartilage materials of the prior art. The
substantial improvement in rate of healing which is obtained from
the use of poly(N-acetylglucosamine), i.e., chitin, as compared to
monomeric N-acetylglucosamine is particularly surprising. As
compared to the great variability in cartilage depending on the
animal, its age and the method of collecting the cartilage, chitin,
particularly chitin of fungal origin, is a relatively uniform and
easily obtained material.
The compositions of the present invention are applied using the
same techniques and processes developed for cartilage, and
N-acetylglucosamine. Thus, it is preferred to topically apply
finely divided chitin directly to the wound surface. However,
tablets, capsules or pellets of chitin may be prepared from
mixtures of chitin, partially depolymerized chitin or chitin
derivatives with well-known pharmaceutical excipients such as
starch, sugar, certain forms of clay, etc. Such tablets, capsules
or pellets may be taken orally or implanted near the situs of the
wound. Alternatively, a colloidal solution may be prepared from
chitin, preferably in isotonic saline, or a water-soluble
derivative of chitin may be dissolved preferably in isotonic saline
solution, and the solution administered intramuscularly,
parenterally or intravenously.
A powder or solution of chitin or of a chitin derivative may also
be used to impregnate a surgical gauze or pad which is applied to
the wound. Chitin may also be dissolved as the alkali chitin
xanthate, spun into fibers and regenerated as the virtually
undegraded polymer in accordance with the procedures described in
the prior art by Thor et al. Partially deacetylated chitin
filaments and fibers may be prepared in accordance with the
procedure described in U.S. Pat. No. 2,040,880. These chitin fibers
may then be used as surgical sutures or included in bandages or
other support base for surgical dressings either in a woven or
nonwoven fabric structure in the manner described in U.S. Pat. No.
3,196,075. Chitin or chitin derivative may also be made up into an
ointment or salve. The use of nonactive carriers for the chitin is
not preferred as the presence of extraneous matter in a wound
frequently tends to interfere with the healing process due to the
interposition effect.
As previously stated, where the chitin is to be applied by
injection, i.e., either intramuscularly, parenterally or
intravenously, it is first necessary to prepare a dispersion or a
solution of the material in a pharmaceutically acceptable liquid.
Colloidal solutions of chitin may be prepared using the method
described by Lingappa and Lockwood in NATURE, 189, page 158 (1961).
When administered intravenously it is preferred to administer the
compound in isotonic solution such as isotonic saline.
The chitin or chitin derivatives may be used alone, in admixture
with each other, with cartilage, or may be co-administered with
other therapeutically effective agents such as ascorbic acid,
ascorbyl palmitate, pharmaceutically acceptable zinc salts such as
zinc oxide, zinc ascorbate, zinc sulfate and zinc stearate;
antiseptics such as thimerosal and benzalkonium chloride; local
anesthetics such as lidocaine and procaine; antibiotics such as
chloramphenicol, sulfanilamide and ampicilline. Combinations of the
therapeutically effective agents described above with chitin and/or
chitin derivatives may be used.
Suitable sources of chitin are from lobsters, shrimp and other
crustacea. To utilize chitin from such sources, it is necessary to
reduce the chitin in particle size to less than about 150 microns
and preferably less than about 50 microns. Due to the tough and
rather fibrous nature of chitin from such sources, this grinding is
difficult and expensive. Accordingly, it is preferred to use chitin
of fungal origin. The cell walls of fungi are made of chitin. It
has been found that it is not necessary to extract the chitin from
the remaining cell material. Thus, if desired, after suitable
sterilization as by heat or gas (i.e., ethylene oxide), the entire
fungal mat produced by fermentation of a fungus in a suitable
nutrient medium may be ground and used to promote healing of
wounds. Preferably, however, the fungal mat is treated to remove
the extraneous materials leaving only the chitin skeletons.
Purifying the material in this manner eliminates the nonchitinous
materials, thus substantially reducing the possibility of an
allergic reaction and eliminating any interference with the healing
process which might be caused by such materials.
Finely divided chitin or chitin derivatives may be applied
topically by blowing a metered amount of the material onto the
wound using a hand atomizer. Alternatively, it may be applied by
dusting as from a hand shaker or may be placed together with an
inert gas under increased pressure (i.e., above atmospheric
pressure) in a pressure vessel. In this latter means of
application, termed "aerosol application," the finely divided
chitin or chitin derivative, optionally with other medicaments as
indicated, may be packaged as a dry aerosol powder as described in
Dutch patent application No. 6,415,252, published July 5, 1965
(this patent application is directed to a medicament for bovine
mastitis but the method of aerosol packaging described is
applicable to powdered medicament having the described particle
size) or as an aerosol foam.
In the following examples, the wound healing efficiency of the
various chitinous materials is determined by using the method of
Prudden et al as described above. In general, at least 10 pairs of
rats are used to obtain a meaningful average for each material
tested. In each of these examples a powder insufflator is used to
apply 2 to 10 mg./cm.sup.2 of wound surface of the material
tested.
EXAMPLE 1
Commercial lobster shell chitin is ground to a fine powder in a
laboratory 4-quart size porcelain jar mill loaded with 1-inch size
(average) flint pebbles in a weight ratio of 1 chitin to 2 pebbles.
Dry ice is then put on top of the mill charge and the mill is kept
open for 5 minutes to allow the CO.sub.2 to displace the air in the
mill. The lid of the mill is then clamped on tight and the grinding
carried out for 96 hours. Approximately 50% of the powdered chitin
passed through a 40 micron screen.
The whole powdered chitin so produced is then applied to the 45
test rats of 45 pairs of rats used in the Prudden et al assay
method described above. The percent of wound healing for the
treated rats, stating the control rats as 100%, is 122%, i.e., the
use of chitin results in an average 22% increase in wound healing
activity.
EXAMPLES 2-5
Various fungi are grown on either brain-heart infusion (200 gm.
calf brain, 250 gm. beef heart, 10 gm. proteose peptone, 2 gm.
dextrose, 5 gm. sodium chloride and 2.5 gm. disodium phosphate)
called "BHI" or on Sabouraud's broth (40 gm. dextrose and 10 gm.
bacto-peptone) called "SAB". The cultures are grown in shallow
layers of media contained in flasks and held stationary until good
growth and extensive sporulation occurs. Prior to collection of the
growth mats, the cultures are killed by placing the flasks into a
closed oven under CO.sub.2 at 127.degree.C. for 3 hours. The flasks
are then cooled in the oven for an additional 1 hour and 15
minutes. Culture broths are removed by filtration through Buchner
funnels and the growth mats washed with distilled water. The mats
are then frozen and lyophilized and the dry products ground in a
mortar with a pestle under CO.sub.2. No attempt is made to purify
the chitin. Twelve pairs of rats are used for each test. Some
inflammation is observed on all treated wounds and infection on
several. The increases in wound healing obtained may be all the
more significant in view of those adverse factors.
______________________________________ % Wound Healing (Control %
Improve- Example Fungus Medium = 100) ment
______________________________________ 2 Mucor spinosus SAB 136 36
3 Aspergillus niger BHI 118 18 4 Penicillium BHI 146 46 5
Cryptococcus BHI 128 28 ______________________________________
EXAMPLE 6
100 grams of dried fungus material (obtained from Penicillium
fungus of Example 4, cultured on a BHI medium, sterilized by
boiling the fungus with the medium and then filtering, washing with
distilled water and drying the fungus material) is defatted by
extracting the solvent-soluble fatty materials with 1,000 ml.
chloroform at room temperature. The chloroform is removed by
filtering and then drying at reduced pressure in a vacuum
desiccator.
The defatted fungus material is treated with 2,000 ml. 1.0 N NaOH
solution for 18 hours at room temperature. The material is then
acidified with HCl. Thereafter the material is dialyzed in
distilled water until the wash water is free from chlorine ions.
This procedure is repeated until a substantially purified material
is obtained. The material is dried in vacuum below 50.degree.C and
is a gray, friable mass.
The dried material is ground in a laboratory mortar and screened
through a 400 mesh standard screen. When the screened material is
applied to 20 test rats of 20 pairs of rats there is obtained an
average of about 25% increase in the wound healing of the treated
rats over the untreated control rats.
EXAMPLE 7
Lobster shell chitin is purified by first slurrying it in 10%
aqueous NaOH for 5 minutes at 80.degree.C, then it is washed,
drained and slurried in 10% HCl for 5 minutes at 80.degree.C,
drained, slurried in water, the pH of the water adjusted to 6 with
dilute aqueous NaOH and finally drained and dried.
The dried chitin material is pulverized to a fineness of about 40
microns. The material shows an average 25% increase in the wound
healing over the untreated control rats.
Although the present invention has been described in conjunction
with preferred embodiments, it is to be understood that
modifications and variations may be resorted to without departing
from the spirit and scope thereof, as those skilled in the art will
readily understand.
* * * * *