Note: Descriptions are shown in the official language in which they were submitted.
WO 92/04896 -1- 2 ~ ~ 9 ~ 1 1 PCT/~S9l/06802
METHOD OF TREATING AUTO-I~IUNE
DISEASES USING G~LLIUM COMPOUNDS
This application i6 a continuation-in-part application
of United States Serial No. 586,491, filed September 21,
1990.
Field of the Invention
This inventlon relates generally to methods of treating
autoimmune diseases, and a method of treatment and
prevention of resistance to transplantation by the use of
gallium or a pharmaceutically acceptable salt thereof.
Backqround of the Invention
Immunosuppressive agents activate or inhibit lymphocyte
proliferation. Lymphocyte proliferation is due to the
interaction between antigens, macrophages, T- and B-
lymphocytes as well as certain chemicals. The presence of
certain antigens may actiYate a particular T- or B-
lymphocyte. Further, certain B-lymphocytes can be activated
by active T-lymphocytes while others are independent of the
T-lymphocyte~ and are activated only by antigens. Activated
T-lymphocytes can cause macrophages to produce a molecule
known as interleuXin-1 which in turn activates both T- and
B-lymphocytes. Activated T-lymphocytes can also produce a
molecule known as interleukin-2 which further induces T-
lymphocyte activation. Additionally, chemicals can trigger
activity in T- or B-lymphocytes. Immunosuppressive agent~
affect the complex interactions between the components o~
the immune system.
The imnune system defends against substances which can
cause disease, however, it cannot distinguish between
helpful and harmful foreign substances and destroys both.
Often times the immunological mechanisms become
sensitized to some part of an individual's own body causing
inter~er2nce with or even destruction of that p rt. The
ability to distinguish between the body'~ own and antigens
not from the body becomes impaired and the body begins to
.. . . .
,
W092/04896
-2- PCT/~lS91/068
destroy itself. The result 1~ an autoimmune disea~e. Some
autoimmune disea~es in man are multiple ~clero~1~, type I
diabetes mellitu6, lupus erythematosus, and Grave~ di~ease.
Suppression of the lmmune systt!m in autoimmune di6eases i~
desirable in minimizing or limiting the aff2ct3 of the
di~ea~e.
Circulating antibodies and cellular immune response~
are involved in the rejection of transplanted tissues and
organs. Unless the donor is the identical twin of the
recipient or is the individual himself, the recipient's
lymphocytes recognize the transplant as not being its own
and immediately responds to destroy it. The exceptions to
this situation ~re transplants to non-vascularized areas,
such as the cornea of the eye, where lymphocytes do not
circulate and therefore are not sensitized ~nd do not prompt
an immune response. It is difficult to suppress the immune
reaction to prevent rejection of the transplant without
severely damaging the patient in other ways.
It has been found that gallium, and gallium nitrate, n
particular, are effective in treating disea6es mediated by
macrophage cell lines, including macrophage/T cell/~ cell
interactions and for the treatment and prevention of
resistance to transplantation.
Gallium ha~ been known for many years to be useful in
the treatment of calcium bone disorders. Gallium is a metal
which belongs to the Group III A Elements of the Periodic
Table. The metallic compounds usad, have, of course, a low
order of tox~city and are pharmaceutically acceptable.
Prior U.S. Patents 4,529,593 issued July 16, 1985 to
Warrell et al; 4,686,104 i~sued August 11, 1987 to Hockman
et al.; and 4,704,277 issued November 3, 1987 to 80ckman et
al. describe methods of preventing exce~sive loss of calcium
from human bone by the ad~inistration of pharmaceutically
acceptable gallium-containing compound~. The '593 patent
teaches the use of pharmaceutically acceptable galllu~ ~alt~
to reduce the exces~lve loss of bone calcium. The patent
spec~fically teaches the use o~ galliu~ to prevent or treat
,
W~92/04~96
-3- PCT/US91/06802
disorder6 a6soc~ated wlth extensive 1065 of calcium from
bons in humans by adminl~tering to the individu~l
pharmaceutlcally acceptable gallium compound. Of specinl
importance among the disorders which, may be thus treated
are hypercalcemia, 06teopenia, osteoporosis, bone
destruction due to metastasi~ from malignant tumors and
hyperparathyroidism. Gallium ~alts which are disclosed to
be of use include nitrate, citrate, and halide, preferably
the chloride, car~on, acetate, tartrate, oxylate, oxide or
hydrated oxide.
Loss of bone mass from increased bone resorption
results in accelerated transfer of calcium into the blood.
This is the major cause of hypercalcemia. Diseases result
when 6ignificant depletion of bone calcium occurs and the
serum calcium level exercises dangerously. The therapeutic
agent o~ choice, according to the aforementioned patent~,
for treating this and many other bone disorder6 iB gallium,
which decreases bone resorption and thereby maintains bone
tissue calcium content.
U.S. Patent 4,303,363 discloses a method of cancer
treatment which uses radioactive 67-gallium as a cytotoxic
agent. -
Heretofore, there has been no link between the
treatment of ~one disorders and cancer by the use of gallium ;-~
and the treatment of autoimmune and immunosuppressant
diseases.
Summar~ of the Inventlon
The present invention relates to methods of tr~ating
macrophage mediated autoimmune diseases, and a method o~
treatment and prevention of resistance to transplantation by
the use of gallium or a pharmaceutically acceptable ~alt
thereof.
~etailed Desc~ion ~f thÇ_p~awi~s
Figures l(a) and l(b) graphically represent the e~fect
of gallium ad~inistration following experimental allergic
- enc2phalitis (EAE) induction;
- ':
'~
, . .. . . . . . . . .. .. . . .
W092/04896 ~i~ ?---
ç~, 4 PCT/US91/06802
Figure 2 graphically repr~sents the effects of galliumto PEAT-specific T line cells during stimulation with PEAT
or Concanavalin A and antigen-presenting cells;
Figure 3 graphically represents a decline in the
expression of I-A in macrophages of BALB/c mice with and
without gallium; and
Figure 4 graphically represents the prevention of
diabetes by the administration of gallium.
Detailed Description of the Invention
In accordance with the present invention, it is
believed that gallium is effective in the treatment of
autoimmune diseases involving various body systems: central
nervous, cardiopulmonary, gastrointestinal, dermatological,
endocrine, renal, reproductive, skeletal and the hepato-biliary
systems. The wide reaching functionality of gallium
indicate that it has promise as ~ therapeutic agent for many
conditions and diseases heretofore not associated with gallium.
More specifically, the present invention relates to the
use of gallium compounds in the treatment of autoimmune
diseases which are mediated by macrophage cell lines
including macrophage/T cell/B cell interaction. These
diseases include multiple sclerosis, thyroiditis, type I
diabetes mellitus, Hashimoto's Disease, Graves Disease,
lupus erythematosus, rheumatoid arthritis, sarcoidosis,
Wegner's granulomatosis and leprosy. In the past, gallium
compounds have been e~ployed in the tr~atment of disorders
associated with bone tissue, and as a cytotoxic agent, but have
not been administered for the treatment of the aforementioned
autoimmune disease~. Further, in the present invention, gallium
is utilized in the suppression of T-cell proliferation.
The following experiments were designed to show the
administration and use of galliu~ for preventing and/or
treating autoimmune diseases and to show the efficacy of
galliuu in the resistance to transplantation. The
experiments are shown in examples which are illustratiYe of
, . . :. . - . , . " ~ .... " .. ., . . . .. , . , . .. . : . ~ ., - ... - ..
W O 92/04896 -5- 2 ~
-- ~ V ~ ~ 1 1 P ~ /US91/068n2
the pre6ent invention. The examples are not lntended to
limit the ~cope of the lnventlon in any way.
E~ampl.e 1
~ ffect ~f q~ um ~dminis~rati~n on expçri~ent~l
allerqiç çncephalomYelitis rE:~F~.
This study demonstrates that the administration of
galllum pravents the development of EAE and results in an
antigen-specific suppression of the lyphocyte proliferative
response. The animal model screens compounds for efficacy
against multiple sclerosis.
Materials and Methods
~ . Male Lewis rats (8-12 wee~s of age) were
purchased from Harland Sprague-Dawley (Indianapolis, IN).
The rats were divided into five groups.
Antiqens. Guinea pig MBP myelin basic protein (GPMEP)
and rat MBP (RMBP) were prepared from 6pinal cords
(Rockland, Inc., Gilbertsville, PA) by cholorform/methanol
extraction. Human MBP (HuMBP) was prepared from cerebral
cortex by cholorform/methanol extraction. The control
antigen ovalbumin (OVA) wa~ obtained from Sigma (St. Loui8,
MO), and ~urified protein derivative (PEAT) was from Parke- -
Davis (Detroit, MI).
Gallium ~itrate. Ben Venue La~oratories, Bedford, Ohio
was the source of a citrated solution of gallium nitrate,
500 mg/20 ml. Rats received in~ections of gallium as
described in Figure la on days 1, 6, 13 and 20 relative to
the induction o~ EAE. Group 1 animal~ received only ~aline;
al~ other6 received weekly injection~ of gallium a~ noted.
Group~ 3 and 5 received saline rather than gallium on day
-1 . '
Induction of EAE. Rats received an intradermal
injection into one hind footpad of guinea plg MBP (25 ug),
combined with complete Freund'~ adjuvant (CFA) on day 0.
- All rats were ~onitored daily for clinical neurol~gic ~igns,
which were scored as ~ollows: limp tail, 1+; ataxi~, 2+;
early or partial paraly~i~, 3~; full hind limb paralysi~, 4+
:
. .-
W092/04896 ~ 6- PCT/US91/06802
deaths. Rats were sacrificed at the ti~e of severe
paralysi~ or 21-38 days after ~;ensitization lf no clinical
- ~igns appeared.
Hlstop,a~h~logi,ç ev~luation. Brains and spinal cords
were removed and fixed in 10~ formalin, and 7-~m transverse
sections of the thalamus, mesencephalon, and cerebellum-pons
and longitudinal sections of the entire spinal cord were
processed for hematoxylin and eosin staining. Histologic
slides were assessed for the presence of perivascular
mononuclear infiltrates and scored as follows: 1-10 lesions,
1+; 11-30 lesions, 2+; and greater than 30 lesions, 3+.
' Results
Figures l(a) and l(b) graphically represent the effect
of gallium administration prior to or following experimental
allergic encephaliti (EAE) induction.
Clinical signs of EAE were ~cored as descri~ed in
Figure l~a~. Rats who received 30 mg per kg on day 6 had
the lowest clinical scores, irrespective of day -1.
CNS Histological Evaluations were performed on all rat~
as r~ported in Bitar et al. ~ell. Immunol. 112:364-370
(1988).
The rats received subcutaneous injections of 30 mg/kg
gallium in tXe form of gallium nltrate beginning six days
after induction of EAE with maintenance doses of 10 mg/kg on
days 13 and 20. Controlled untreated rats progressed to
maximu~ paralysis by day 13 as ~hown in Figure l(a), whereas
gallium treated rats exhibited either no sign~ or ~inimal
disease (partial left tail). Studies were subsequently
undertaken to determine the ef~ect~ of dosage~ and t1ming of
administration gallium on EAE. Gallium totaling 0-80 mg/kg
was ac~inistered at weekly intervals beginning either be~ore
or after induction of EAE as ~hown in Table 1. Mark~d
suppr~ssion of EAE was demonstrated in all gallium treated
(30-R0 mg/kg) animals. Maximum inhibition of clinical
di~ease was ach~eved when 30 mg/kg was given on day 6, ancl
~aximal ~uppression of histopatholo~i~ chang~ occurred at
the larg~st close of galliu~. No demonstrable adverse
W092/04~96 7~ ~ PCT/U~91/068~2
clinical effect~ of gallium were notPd in any of thQ in YiYQ
BtUdieE~ .
The optimum timing for admini~tration ~f galllu~
following induction of EAE was determlned by injectlng 30
mgtkg as a single dose to the rats on day 3, 6, 9, or 12
after induction. Figure l(b) ~hows that gallium exerts its
maximal suppressive effect when given on day 6, with a
significant inhibitory effect also observable when
administered on day 3 or 9. ]?ollowing compl~te recovery,
rats from each group were reinjected with myelin ~a6ic
protein (MEP) on day 28 and none developed EAE.
ExamPle 2
In vitro Study of the Effect of -
Gallium upon LYmphoid Cells
Materials and Method
Rats. Thirty-eight male Lewis rats (135-195 g) were
purchased from Charles River La~oratories (Portage, MI) for
use in this experiment.
Esta~lish~ent of a Purified pro~ein deriY~tiye L~EAT!
specifiç T l~mPhocvte line. A specific PEAT-reactive T-cell
line was prepared by conventional techniques. Cells
(8xlO6/ml) from lymph nodes of CFA-immuni2ed Lewi~ rats were
incubated for 3 days in tissue culture Petri dishes in RP~I
1640 (Whittaker M.A. Bioproducts, Walkersville, MD)
containing 50 Utml penicillin, 50 ~g/ml streptomycin, 50 ~
2-mercaptoethanol, 2 mM gluta~ine, 26 mM ~epes~ 1~ fresh
autologous rat serum, and 40 ~g/ml PEAT. Lymphobla&ts were
obtained by centrifugation, using lymphocyte-separat~ng
medium (Organo~ Teknika, Durha~, NC) (240 g, 25 min~, and
the cells were washed and cultured in medium containing 10%
fetal bovine serum (Whittaker M.A. Bioproducts) and 10%
~/v) rat T-cell growth factor (24-hr super~atant from rat
splee~ CellB stimulated with Concanav~lin A). After 4-8
days in culture, the cells were restimulated ~or 3 day~ wit~ -
PEAT (20 ~g/~l) in the presence o~ ~ynerglc ~-~rradiated
.
W092/04896
-8- PCT/USgl/068~2
(3300 r) thymocytes (107tml) as a ~ource of antigen
presenting cells. The T-line cells werQ alternately
expanded in rat growth factor or restimulated with antigen
and irradiated thymocytes. After the ~econd round o~
antigen restimulation, ~n vitro lymphocyte prollferative
assays revealed a PEAT-specific response. After
establishment of the T-cell line, restimulation with antigen
was performed at approximately two-week interval~.
Procedurç
To measure the effect of Ga on the PEAT-specific T-cell
line, a lymphocyte proliferative assay was used with
[3H]thymidine incorporation as the endpoint. T cells
(2xlO4/well) were cultured in 150 ~l of RPMI complete medium
together with 10~/well) were cultured in 150 ~l of RPMI
complete medium together with 106 irradiated (3300 r)
thymocyte~ and 20 ~g/ml PD or 2 ~g/ml Concanavalin A in 96-
- well round-bottomed plates (Flow, Maclean, VA). Cultures
were incubated for a total of 72 hr in 7% CO2 at 37C in a
humidified atmosphere and pulsed with [~]thymidine (1
l~Ci/well) (Amersham, Arlington Heights, IL) during the
final 18 hr of culture. Cells were harvested using a semi-
automatic sample harvester (Skatron, Sterling, YA) and
counted by liquid ~cintillation. Cultures contained either
no Ga or Ga nitrate at 1.0 mg/ml. In order to control for
nonspecific toxicity of the Ga preparation, an aliquot of T
cell~ wa~ incubated with Ga (1.0 mg/ml) for 1 hr and washed
prior to culture.
R~sults
Figure 2 ~hows the effects of addition of gallium to
PEAT-~pec~fic ~ line cell~ during stimulation with PEAT or
Concanavalin A and a~tigen-presenting cells.
~a~Ple 3
Ex~ession of Maior Histocompatibility
ÇQmplex Clas~ II Glycop~o~eins.
Sixty-four male BALB/c mice received an i.p. ln~Pction
of 2 ml o~ 3~ thiogly olate medium in ~terile water to
'.
W092/04896 -9~ PCT/U~91/06~02
~nduce migration of macrophage~ to the peritoneum. A~ter 3-
5 day6, the mice were euthanized and 15-20 x 1o6 cell~
coll~cted from the peritoneum of e~ch by lavage with ~ank~'
balanced ~alt ~olution (Wh:Lttaker M.A. Bioproduct~).
Viability, determlned by trypan blue ~xclu6ion, wa~ always
greater than 95~. The cell~ were added to Dulbecco's
modified Eagle's medium, supplemented with 10% fetal bovine
serum (Hyclone Laboratories, Lcgan, UT), 2 mM glutamine, 100
U of penicillin, and lOO ~g of 6treptomycin, to a final
concentration of 2 x 105 cells/ml for each mouse. One-half
milliliter of each cell suspension was added to chambers of
multichambered Lab-Tek slides (Nunc, Inc., Naperville, IL).
Macrophages were allowed to adhere to the slides for 24 hr;
then nonadherent cells were removed by washing with Hanks'
balanced salt solution. The macrophages were incubated for
48 hr with lOO U/well of murine recombinant ~-interferon
(Amgen, Thousand Oaks, CA). A macrophage-activating factor, :
~-interferon stimulates the expression of I-~ (an MHC clas~
II glycoprotein) and enhances the cell'~ ability to kill
microorganisms and tumor cell~. Gallium nitrate (0 ~A), 05
(B), 1.0 (C), or 2.5 mg (D)) was subsequently added to each
well, and the percentage of cells expressing I-A was
determined at 1, 4, 8, and 16 hr by indirect fluorescence,
using a UV microscope (Zeiss, Oberkochen, W. Germany~
Specifically, the macrophages were treated with 5% rabbit
~erum at 4C to block Fc receptor binding. After 30 ~inutes
the cell6 wero washed and then incubated with 0.5 ~g of
monoclonal anti I-Ad (MKD-6, American Type Cultura
Collection, Rockville, MD), grown in the same medium as ~he
macrophages. MXD-6 i9 a cell line which produce~ cytotoxlc
monoclonal antibodies which react with murine I-Ad antigen.
After 40 ~in 'at 4C) he ~ell~ ~ere washed with ~ank~', and
FITC-conjugated goat anti~ouse F(ab' )2 immunoglobulin
(Organon Teknika) was added for 40 minO This wa~
su~sequently washed off, the tops of the slides removed, a
W092/04896 ~ 1
- o- PCT/US91/06802
coverslip placed over ~he cells, and the number of
fluorescent cells assessed ln a blinded fashion.
Rç~lts
Figure 3 ~hows a decline in the expresslon of I-A when
peritoneal macrophages of BALB/c mice were incubated with
gallium. This lnhibition of expression diminished with
time, ranging from approximately 45S at 1 hr to 0~ (full
recovery) by 16 hr. Cells remained viable throughout the
study.
Exam~le 4
Treatment of Pre-Diabetic Non-Obes Q (NOD) Mice
with Gallium Nitrate Prevents Development of Diabetes.
Diabetogenesis in NOD/mice is an immune media~ed
disease, with generation of auto-immune reactivity against
pancreatic ~ cells requiring participation of macrophages
and T lymphocytPs. In the specific pathogen-free NOD/LT
colony, untreated virgin females exhibited a 79~ diabetes
incidence by eight months of age compared to a 45S incidence
in virgin males. Diabetes in NOD mice can be circumvented
by a variety of immunomodulatory procedures initiated
shortly after weaning. These include immunosuppressive
treatment6 impairing viability and/or function of either
macrophages or T cells. In this Example, gallium nitrate ifi
used to circumvent diabetes.
Materials and ~ethods
NOD mice. 24 NOD/Lt female mice from litters born
between 12-l-90 and 12-6-90 from Jackson Laboratories, Bar
Harbour, Maine were randomly sorted into two group of
twelve. Mice were caged threej~ide ln double penned plastic
boxe~ with a natural ingredient diet (old Guilford 96 w) and
Libitum.
Gallium Nitrate. A citrated solution of gallium
nitrate per 500 mg/20 ml was obtained from Ben Venue
- Laboratories, Bedford, Ohio. Lot No. 89/215 wa3 used in
this experiment.
W~92/04896 ~ PC~/US91/06802
P~roçedures
At BiX weeks of age, twelve female~ received an
in~ection i.p. of 45 mg/kg body weight followed by once
weekly injection~ i.p. of 45 mg/kg body weight. The
controlled group of twelve females received an in~ection o~
vehicle (citrated sallne ~olution) at the same time~. Mice
were weighed and checked for glycosuria using TES-tape
diagnostic tapes supplied by Eli Lilly and Company at weekly
interval~. A diagno~is of diabetes was made if the mice
remained glycosuria for three consecutive weeks and lost
weight. Diabetic mice were euthanized without further
analysis. At the end of 20 weeks, weekly injections of
gallium nitrate in vehicle were discontinued; the mice were
aged at 30 weeXs to determine whether absence of disea e
would persist without treatment.
~esul~S
NOD/LT females after 14 weeks of gallium injections did
not show reduced body weights compared to vehicle control~
(mean + SEM = 23.0 ~ O.7 g (n - l0) for gallium recipient~
versus 23.0 + 0.5 (n = 8) for vehicle controls. Two female~
in the gallium treatment group had to be deleted from the
experiment due to development of malocculsions and
subsequent weight loss. Malocculsions are detected
sporadically in the NOD/Lt colony and are not considered to
~e direct effects of the gallium treatment.
Figure 4 depicts the diabetes incidence in the mice.
The ability of gallium nitrate injections to circumvent
development of clinical diabete6 is unequivocal. Thare ar~
no ca~e~ of overt diabete in the group during the periQd o~
weekly qallium administration. Further, there wa~ no
"breakthrough" of diabetes within a 5 week period a~ter
cessation of gallium injections. In distinctio~, ~ sh~rp
increase in c1iabetes incidence occurred in vehicle controls
during the ~ame period (21-25 week6).
WO92/04896 ~ 12- PCr/~'S91/0~80'
Example 5
Trans~lan~a~i~n Q~ MQuse---Heart
The heart of a DBA/2 from Harland mouse waR removed.
The abdomen of a C57/BL/6 mouse from Sprague Dawley,
Indianapolls, IN. was opened and the a~dominal aorta
displayed. The donor heart was transplanted by attaching
the donor aorta to the recipient abdominal aorta and the
donor pulmonary artery to the recipient vena cava. A single
dose of 45 mg/kg of elemental gallium was administered
subcutaneously after the surgical procedure was completed.
The transplanted heart remained viable for eleven days. A
controlled experiment in a mouse from the same strain and
source using the foregoing procedure without the
administration of gallium nitrate allowed the heart to
remain viable for only five days.
As used herein, the term "patient" refers to a war~
blooded animal such as a mammal which i5 affected with a
disease, such as an autoimmune disease, or is in danger of
rejection of a transplanted tissue or organ. It i~
understood that human~ are included within the scope of the
term "patient. n Based on standard clinical and laboratory
tests and procedures, an attending diagnostician, a~ a
person skilled in the art, can readily identify those
patients who are in need of treatment with immunosuppre6sive
agents.
According to the present invention, in order to obtain
the beneficial effects of gallium in treating autoimmune
diseases and resi~tance to transplantation, pharmacautically
acceptable gallium containing co~pounds ~re ad~inistered to
the patient i~ an amount sufficient to provide therapeu~ic ~ -
level3 of gallium. Therapeutio level~ are obtained when
gallium is present in a steadyi6tate concentration in blood.
Typically, the amount of elemental gallium administered is
from about 0.05 to about 50 mg/per kg per day of ~ody
weight. Preferably, this amount range~ fro~ about 0.05 tD
about .5 mg/per kg of body weight~
~092/Oq896 -13~
3 1 P~/l,'S91/06802
Galli~m containlng compound~, effective wlth this
invention, may b~ any gallium containing compound~ ln non-
nephrotoxic amount~ to lnhibit or treat autoimmune disea~e~,
and re6i~tanc~ to transplantation, in patients 6uffering
from the aforementioned condition~ by administering to a
patlent a therapeutically effective amount of such a
compound. Preferably, the compounds may be selected ~rom
the group consisting of gallium nitrate, gallium citrate,
gallium chloride, gallium carbonate, gallium acetate,
gallium lactate, gallium tartrate, gallium oxalate, gallium
oxide and hydrated gallium oxide.
Gallium containing compounds are useful in formulations
having a variety of routes of administration. The route(s)
of administration useful in a particular application for the
treatment of autoimmune diseases, immuno~uppression and
resistance to transplantation is apparent to one 6killed in
the art. Routes of administration include but no~ llmlted
to topical, transdermal, parenteral, transbronchial and
transalveolar.
Formulations of gallium contalning compounds 6uitable
for topical application include, but are ~ot l:Lmited to,
implants; ointments, creams, resin~ and gels. For~ulation~
suitable for transdermal application include, but are not
limited to, suspensions, oils, creams and ointments applied
directly or attached to a protective carrier such a~ a
patch. Formulations suitable for parenteral administration
include, but are not limited to, sterile solution6 for
intravenous, intramuscular or ~ubcutaneous iniection.
Formulations suitable for transbronchial and tran6alveolar
administration include, but are not li~ited to, various
types of aerosol~ for inhalation. The above-mentioned
formulations are meant to describe but not limit the ~ethod~
of admini~tering gallium containing compound~. The m~thod~
of making the various formulation~ are within the ability of
one skilled i~ the art and need not be described in detail.
.
The terms and expressions which had been ~mployed are
used as ter~s of description and not of llmitation, ~nd
W~92/04896 ~ 14-
~ ~ , PCT/US91/06802
thQre i5 no intention in the U8Q of such term~ and
expr~slon~ o~ excluding any Qquiv~lents of the feature
shown and described or portions thereof, it being
recognized that variou~ modifications are possible within
the scope o~ the inventlon.
