U.S. patent application number 14/478519 was filed with the patent office on 2015-03-05 for use of lipid conjugates in the treatment of diseases or disorders of the eye.
The applicant listed for this patent is CELSUS THERAPEUTICS PLC, YISSUM RESEARCH DEVELOPMENT COMPANY. Invention is credited to YUVAL COHEN, SHAUL YEDGAR.
Application Number | 20150065467 14/478519 |
Document ID | / |
Family ID | 39402091 |
Filed Date | 2015-03-05 |
United States Patent
Application |
20150065467 |
Kind Code |
A1 |
YEDGAR; SHAUL ; et
al. |
March 5, 2015 |
USE OF LIPID CONJUGATES IN THE TREATMENT OF DISEASES OR DISORDERS
OF THE EYE
Abstract
In one embodiment, the invention provides a method of treating,
reducing the incidence, reducing the severity or pathogenesis of an
eye disease or disorder in a subject, including, inter alia,
retinal detachment, macular degeneration, glaucoma or retinopathy,
comprising the step of administering an effective amount of a lipid
or phospholipid moiety bound optionally via a spacer to a
physiologically acceptable monomer, dimer, oligomer, or polymer via
an ester or amide bond, and/or a pharmaceutically acceptable salt
or a pharmaceutical product thereof. This invention also provides a
contact lens solution comprising a lipid or phospholipid moiety
bound optionally via a spacer to a physiologically acceptable
monomer, dimer, oligomer, or polymer via an ester or amide bond,
and/or a pharmaceutically acceptable salt or a pharmaceutical
product thereof.
Inventors: |
YEDGAR; SHAUL; (JERUSALEM,
IL) ; COHEN; YUVAL; (BROOKLYN, NY) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
YISSUM RESEARCH DEVELOPMENT COMPANY
CELSUS THERAPEUTICS PLC |
JERUSALEM
MAYFAIR |
|
IL
GB |
|
|
Family ID: |
39402091 |
Appl. No.: |
14/478519 |
Filed: |
September 5, 2014 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
11984223 |
Nov 14, 2007 |
8865681 |
|
|
14478519 |
|
|
|
|
Current U.S.
Class: |
514/114 |
Current CPC
Class: |
A61K 31/726 20130101;
A61P 27/06 20180101; A61K 31/718 20130101; A61K 47/544 20170801;
A61K 47/56 20170801; A61K 31/717 20130101; Y02A 50/401 20180101;
A61K 31/685 20130101; A61K 31/715 20130101; A61P 27/02 20180101;
A61K 31/661 20130101; A61K 9/0048 20130101; A61K 47/61 20170801;
A61K 31/727 20130101 |
Class at
Publication: |
514/114 |
International
Class: |
A61K 31/661 20060101
A61K031/661; A61K 47/48 20060101 A61K047/48 |
Claims
1. A method of treating a disease or disorder of the eye in a
subject comprising the step of contacting said subject with a
compound comprising a lipid or phospholipid moiety bound optionally
via a spacer to a physiologically acceptable polymer, and/or a
pharmaceutically acceptable salt or a pharmaceutical product
thereof.
2. The method according to claim 1, wherein said phospholipid
moiety is phosphatidylethanolamine.
3. The method according to claim 2, wherein said
phosphatidylethanolamine is dipalmitoyl
phosphatidylethanolamine.
4. The method according to claim 2, wherein said
phosphatidylethanolamine is dimyristoyl
phosphatidylethanolamine
5. The method according to claim 1, wherein said physiologically
acceptable monomer, dimer, oligomer, or polymer is polygeline.
6. The method according to claim 1, wherein said physiologically
acceptable monomer, dimer, oligomer, or polymer is a
polypyranose.
7. The method according to claim 6, wherein said polypyranose is
carboxymethylcellulose.
8. The method according to claim 6, wherein said polypyranose is
alginate.
9. The method according to claim 6, wherein said polypyranose is
hydroxyethyl starch.
10. The method according to claim 1, wherein the lipid or
phospholipid moiety bound optionally via a spacer to a
physiologically acceptable monomer, dimer, oligomer, or polymer via
an ester or amide bond, and/or a pharmaceutically acceptable salt
or a pharmaceutical product thereof is represented by the structure
of the general formula (A): ##STR00054## wherein L is a lipid or a
phospholipid; Z is either nothing, ethanolamine, serine, inositol,
choline, phosphate, or glycerol; Y is either nothing or a spacer
group ranging in length from 2 to 30 atoms; X is a physiologically
acceptable polymer; and n is a number from 2 to 1000.
11. The method of claim 10, wherein L is phosphatidyl, Z is
ethanolamine, Y is nothing, and X is carboxymethylcellulose or a
glycosaminoglycan.
12. The method of claim 10, wherein the phosphatidylethanolamine
moiety is dipalmitoyl or dimyristoyl phosphatidylethanolamine.
13. The method of claim 10, wherein the lipid or phospholipid
moiety bound optionally via a spacer to a physiologically
acceptable monomer, dimer, oligomer, or polymer via an ester or
amide bond, and/or a pharmaceutically acceptable salt or a
pharmaceutical product thereof is represented by the structure of
the general formula (I): ##STR00055## wherein R.sub.1 is a linear,
saturated, mono-unsaturated, or poly-unsaturated, alkyl chain
ranging in length from 2 to 30 carbon atoms; R.sub.2 is a linear,
saturated, mono-unsaturated, or poly-unsaturated, alkyl chain
ranging in length from 2 to 30 carbon atoms; and Y is either
nothing or a spacer group ranging in length from 2 to 30 atoms; X
is a physiologically acceptable polymer; and n is a number from 1
to 1000.
14. The method of claim 13, wherein n is a number from 2 to
100.
15. The method of claim 10, wherein the lipid or phospholipid
moiety bound optionally via a spacer to a physiologically
acceptable monomer, dimer, oligomer, or polymer via an ester or
amide bond, and/or a pharmaceutically acceptable salt or a
pharmaceutical product thereof is represented by the structure of
the general formula (III): ##STR00056## wherein R.sub.1 is a
linear, saturated, mono-unsaturated, or poly-unsaturated, alkyl
chain ranging in length from 2 to 30 carbon atoms; R.sub.2 is a
linear, saturated, mono-unsaturated, or poly-unsaturated, alkyl
chain ranging in length from 2 to 30 carbon atoms; Z is either
nothing, inositol, choline, or glycerol; Y is either nothing or a
spacer group ranging in length from 2 to 30 atoms; X is a
glycosaminoglycan; and n is a number from 1 to 1000.
16. The method of claim 15, wherein n is a number from 2 to
100.
17. The method of claim 10, wherein the lipid or phospholipid
moiety bound optionally via a spacer to a physiologically
acceptable monomer, dimer, oligomer, or polymer via an ester or
amide bond, and/or a pharmaceutically acceptable salt or a
pharmaceutical product thereof is represented by the structure of
the general formula (IV): ##STR00057## wherein R.sub.1 is either
hydrogen or a linear, saturated, mono-unsaturated, or
poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon
atoms; R.sub.2 is a linear, saturated, mono-unsaturated, or
poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon
atoms; Z is either nothing, inositol, choline, or glycerol; Y is
either nothing or a spacer group ranging in length from 2 to 30
atoms; X is a glycosaminoglycan; and n is a number from 1 to
1000.
18. The method of claim 17, wherein n is a number from 2 to
100.
19. The method of claim 10, wherein the lipid or phospholipid
moiety bound optionally via a spacer to a physiologically
acceptable monomer, dimer, oligomer, or polymer via an ester or
amide bond, and/or a pharmaceutically acceptable salt or a
pharmaceutical product thereof is represented by the structure of
the general formula (V): ##STR00058## wherein R.sub.1 is a linear,
saturated, mono-unsaturated, or poly-unsaturated, alkyl chain
ranging in length from 2 to 30 carbon atoms; R.sub.2 is either
hydrogen or a linear, saturated, mono-unsaturated, or
poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon
atoms; Z is either nothing, inositol, choline, or glycerol; Y is
either nothing or a spacer group ranging in length from 2 to 30
atoms; X is a glycosaminoglycan; and n is a number from 1 to
1000.
20. The method of claim 19, wherein n is a number from 2 to 100.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is continuation of U.S. application Ser.
No. 11/984,223, filed Nov. 14, 2007, which claims the benefit of
U.S. Provisional Application Ser. No. 60/858,706, filed Nov. 14,
2006 and U.S. Provisional Application Ser. No. 60/907,785, filed
Apr. 17, 2007. All applications above are incorporated herein in
their entirety.
FIELD OF THE INVENTION
[0002] This invention provides compounds and methods of use thereof
for reducing the incidence, reducing the severity or pathogenesis,
or treating a disease or disorder of the eye in a subject,
including, inter alia, retinal detachment, macular degeneration,
glaucoma or retinopathy, as well as contact lens solutions
comprising said compounds.
BACKGROUND OF THE INVENTION
[0003] Compounds for use in the present invention are thought to
inhibit the enzyme phospholipase A2 (PLA2, EC 3.1.1.4).
Phospholipase A2 catalyzes the breakdown of phospholipids at the
sn-2 position to produce a fatty acid and a lysophospholipid. The
activity of this enzyme has been correlated with various cell
functions, particularly with the production of lipid mediators such
as eicosanoid production (prostaglandins, thromboxanes and
leukotrienes), platelet activating factor and lysophospholipids.
Compounds for use in the present invention may offer a wider scope
of protection of cells and organisms from injurious agents and
pathogenic processes, including the prevention and treatment of eye
diseases.
[0004] The elderly population in the United States is increasing
rapidly. By the year 2030, approximately 70 million Americans will
be over 65 years of age. Loss of vision among the elderly is a
major health care problem: approximately one in three elderly
persons has some form of vision-reducing eye disease by the age of
65. Vision impairment is associated with a decreased ability to
perform activities of daily living and an increased risk for
depression.
[0005] Although estimates vary, there are approximately 10 million
blind and visually impaired people in the United States, of which
approximately 5.5 million are elderly individuals. Cataract,
glaucoma, age-related macular degeneration, and diabetic
retinopathy are the four eye disorders that pose the greatest
threats to vision after age 40. Patients with age-related macular
degeneration often have the following symptoms: blurred vision,
image distortion, central scotoma, and/or difficulty reading;
Patients with glaucoma often have the following symptoms: visual
field loss and/or blurred vision (late); Patients with cataracts
often complain of blurred vision, glare, and/or monocular diplopia;
Patients with diabetic retinopathy often have the following
symptoms: Bluffed vision, floaters, visual field loss, often have
poor night vision.
[0006] Other common eye disorders of aging, include presbyopia, dry
eye, floaters and flashes, retinal detachment, and eyelid problems
such as drooping upper or lower lids.
[0007] In the United States, diabetes is responsible for 8% of
legal blindness, making it the leading cause of new cases of
blindness in adults 20-74 years of age. Each year, between 12,000
to 24,000 people lose their sight because of diabetes, making
patients with diabetes 25 times more likely to lose vision than
those who are not diabetic, according to the American Academy of
Ophthalmology. In addition, diabetic retinopathy often leads to
additional eye disorders such as retinal detachment, glaucoma
cataract, and corneal disease, contributing to the high rate of
blindness in diabetics.
[0008] Over 29 million people in the United States wear contact
lenses, which provide a safe and effective way to correct vision
when used with care and proper supervision. However, many contact
lens wearers and potential contact lens wearers suffer from
discomfort, dry eyes, and infection as a result of contact lens
use. There is therefore a need to design contact lenses to comprise
compounds that allow the lens to be more biocompatible,
comfortable, tear-wettable, anti-bacterial and oxygen
permeable.
SUMMARY OF THE INVENTION
[0009] In one embodiment, the invention provides a method of
reducing the incidence, reducing the severity or pathogenesis of a
disease or disorder of the eye in a subject comprising the step of
contacting said subject with a compound comprising a lipid or
phospholipid moiety bound optionally via a spacer to a
physiologically acceptable monomer, dimer, oligomer, or polymer via
an ester or amide bond, and/or a pharmaceutically acceptable salt
or a pharmaceutical product thereof. In another embodiment, the
invention provides a method of treating a disease or disorder of
the eye in a subject comprising the step of contacting said subject
with a compound comprising a lipid or phospholipid moiety bound
optionally via a spacer to a physiologically acceptable monomer,
dimer, oligomer, or polymer via an ester or amide bond, and/or a
pharmaceutically acceptable salt or a pharmaceutical product
thereof. In another embodiment, the invention provides a contact
lens solution comprising a lipid or phospholipid moiety bound
optionally via a spacer to a physiologically acceptable monomer,
dimer, oligomer, or polymer via an ester or amide bond, and/or a
pharmaceutically acceptable salt or a pharmaceutical product
thereof.
[0010] In one embodiment, the compound for use in the solutions,
compositions and methods of the present invention is represented by
the structure of the general formula (A):
##STR00001##
[0011] wherein [0012] L is a lipid or a phospholipid; [0013] Z is
either nothing, ethanolamine, serine, inositol, choline, phosphate,
or glycerol; [0014] Y is either nothing or a spacer group ranging
in length from 2 to 30 atoms; [0015] X is a physiologically
acceptable monomer, dimer, oligomer, or polymer; and [0016] n is a
number from 1 to 1000; [0017] wherein any bond between L, Z, Y and
X is either an amide or an esteric bond.
[0018] In one embodiment, the compound for use in the solutions,
compositions and methods of the present invention is represented by
the structure of the general formula (A):
##STR00002##
[0019] wherein [0020] L is a lipid or a phospholipid; [0021] Z is
either nothing, ethanolamine, serine, inositol, choline, phosphate,
or glycerol; [0022] Y is either nothing or a spacer group ranging
in length from 2 to 30 atoms; [0023] X is a physiologically
acceptable monomer, dimer, oligomer, or polymer; and [0024] n is a
number from 2 to 1000; [0025] wherein any bond between L, Z, Y and
X is either an amide or an esteric bond.
[0026] In another embodiment, the compound for use in the
solutions, compositions and methods of the present invention is
represented by the structure of the general formula (I):
##STR00003##
wherein [0027] R.sub.1 is a linear, saturated, mono-unsaturated, or
poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon
atoms; [0028] R.sub.2 is a linear, saturated, mono-unsaturated, or
poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon
atoms; [0029] Y is either nothing or a spacer group ranging in
length from 2 to 30 atoms; [0030] X is alginate,
hydroxyethylstarch, polygeline, carboxymethylcellulose, or a
combination thereof; and [0031] n is a number from 1 to 1000 [0032]
or [0033] wherein [0034] R.sub.1 is a linear, saturated,
mono-unsaturated, or poly-unsaturated, alkyl chain ranging in
length from 2 to 30 carbon atoms; [0035] R.sub.2 is a linear,
saturated, mono-unsaturated, or poly-unsaturated, alkyl chain
ranging in length from 2 to 30 carbon atoms; [0036] Y is either
nothing or a spacer group ranging in length from 2 to 30 atoms;
[0037] X is alginate, hydroxyethylstarch, polygeline,
carboxymethylcellulose, or a combination thereof; and [0038] n is a
number from 1 to 1000.
[0039] In another embodiment, the compound for use in the
solutions, compositions and methods of the present invention is
represented by the structure of the general formula (I):
##STR00004##
wherein [0040] R.sub.1 is a linear, saturated, mono-unsaturated, or
poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon
atoms; [0041] R.sub.2 is a linear, saturated, mono-unsaturated, or
poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon
atoms; [0042] Y is either nothing or a spacer group ranging in
length from 2 to 30 atoms; [0043] X is alginate,
hydroxyethylstarch, polygeline, carboxymethylcellulose, or a
combination thereof; and [0044] n is a number from 2 to 1000 [0045]
or [0046] wherein [0047] R.sub.1 is a linear, saturated,
mono-unsaturated, or poly-unsaturated, alkyl chain ranging in
length from 2 to 30 carbon atoms; [0048] R.sub.2 is a linear,
saturated, mono-unsaturated, or poly-unsaturated, alkyl chain
ranging in length from 2 to 30 carbon atoms; [0049] Y is either
nothing or a spacer group ranging in length from 2 to 30 atoms;
[0050] X is alginate, hydroxyethylstarch, polygeline,
carboxymethylcellulose, or a combination thereof; and [0051] n is a
number from 2 to 1000.
DETAILED DESCRIPTION OF THE INVENTION
[0052] In one embodiment, the invention provides a method of
reducing the incidence, reducing the severity or pathogenesis of a
disease or disorder of the eye in a subject comprising the step of
contacting said subject with a compound comprising a lipid or
phospholipid moiety bound optionally via a spacer to a
physiologically acceptable monomer, dimer, oligomer, or polymer via
an ester or amide bond, and/or a pharmaceutically acceptable salt
or a pharmaceutical product thereof.
[0053] In one embodiment, the invention provides a method of
suppressing or inhibiting, a disease or disorder of the eye in a
subject, comprising the step of contacting a cell with a compound
comprising a lipid or phospholipid moiety bound to a
physiologically acceptable monomer, dimer, oligomer, or polymer,
and/or a pharmaceutically acceptable salt or a pharmaceutical
product thereof.
[0054] In another embodiment, the invention provides a method of
treating a disease or disorder of the eye in a subject, comprising
the step of contacting said subject with a compound comprising a
lipid or phospholipid moiety bound optionally via a spacer to a
physiologically acceptable monomer, dimer, oligomer, or polymer via
an ester or amide bond, and/or a pharmaceutically acceptable salt
or a pharmaceutical product thereof.
[0055] In another embodiment, the invention provides a method of
preventing a disease or disorder of the eye in a subject,
comprising the step of contacting said subject with a compound
comprising a lipid or phospholipid moiety bound optionally via a
spacer to a physiologically acceptable monomer, dimer, oligomer, or
polymer via an ester or amide bond, and/or a pharmaceutically
acceptable salt or a pharmaceutical product thereof.
[0056] In one embodiment, the invention provides for the use of a
lipid or phospholipid moiety bound optionally via a spacer to a
physiologically acceptable monomer, dimer, oligomer, or polymer via
an ester or amide bond, in the preparation of a composition for
suppressing, inhibiting, preventing or treating a disease or
disorder of the eye in a subject. In another embodiment, the
invention provides for the use of a lipid or phospholipid moiety
bound optionally via a spacer to a physiologically acceptable
monomer, dimer, oligomer, or polymer via an ester or amide bond, in
the preparation of a composition for reducing the incidence,
reducing the severity or pathogenesis of a disease or disorder of
the eye in a subject.
[0057] In one embodiment, the term "a disease or disorder of the
eye" refers to any one or more of the following conditions: retinal
detachment, macular edema, retinopathy, age-related macular
degeneration, macular cyst, macular hole, solar retinopathy,
diabetic retinopathy, branch retinal vein occlusion, or Lebers
congenital amaurosis. In another embodiment, the term "a disease or
disorder of the eye" refers to any one or more of the following
conditions: corneal graft rejection, uveitis, inflammatory eye
diseases, infectious eye diseases, ocular tumours, neovascular
proliferative diseases, neovascular maculopathies, rheumatoid
corneal melting disorders, or autoimmune disorders.
[0058] It is to be understood that the method of the present
invention may be used to prevent or treat any disorder or disease
of the eye or associated with the eye, or in another embodiment,
any ophthalmic disorder. In one embodiment, the methods of the
present invention may be used to prevent, suppress, inhibit or
treat episcleritis, scleritis, or a combination thereof. In another
embodiment, the methods of the present invention may be used to
prevent, suppress, inhibit or treat retinopathy, including, inter
alia, diabetic retinopathy, glaucoma, macular degeneration, retinal
detachment, or a combination thereof. In another embodiment, the
methods of the present invention may be used to prevent, suppress,
inhibit or treat any one or more of the following diseases or
disorders, or symptoms as a result thereof: achromatopsia/Maskun,
amblyopia, anisometropia, Argyll Robertson pupil, astigmatism,
anisometropia, blindness, chalazion, color blindness,
achromatopsia/Maskun, esotropia, exotropia, floaters, vitreous
detachment, Fuchs' dystrophy, hypermetropia, hyperopia,
hypertensive retinopathy, iritis, keratoconus, Leber's congenital
amaurosis, Leber's hereditary optic neuropathy, macular edema,
myopia, nyctalopia, ophthalmoplegia, including progressive external
ophthalmoplegia and internal opthalmoplegia, opthalmoparesis,
presbyopia, pterygium, red eye (medicine), retinitis pigmentosa,
retinopathy of prematurity, retinoschisis, river blindness,
ophthalmoplegia, scotoma, snow blindness/arc eye, eyelid disorders,
ptosis, extraocular tumours, strabismus, which in one embodiment is
esotropias, exotropias, vertical patterns, eye injuries, or a
combination thereof. In another embodiment, the methods of the
present invention may be used to prevent, suppress, inhibit or
treat any one or more of the following diseases or disorders, or
symptoms as a result thereof: neovascular glaucoma, retrolental
fibroplasias, Vitamin A deficiency, contact lens overwear, atopic
keratitis, superior limbic keratitis, pterygium keratitis sicca,
Sjogrens syndrome, acne rosacea, phylectenulosis, syphilis, lipid
degeneration, chemical burns, Mooren ulcer, Terrien's marginal
degeneration, marginal keratolysis, polyarteritis, trauma, Wegeners
sarcoidosis, scleritis, Steven's Johnson disease, periphigoid
radial keratotomy, sickle cell anemia, syphilis, pseudoxanthoma
elasticum, Paget's disease, vein occlusion, artery occlusion,
carotid obstructive disease, chronic uveitis/vitritis, Lyme
disease, systemic lupus erythematosis, Eales disease, Behcet's
disease, presumed ocular histoplasmosis, Best's disease, optic
pits, Stargardt's disease, pars planitis, chronic retinal
detachment, retinoschisis, hyperviscosity syndromes, toxoplasmosis,
trauma, post-laser complications, rubeosis, or a combination
thereof.
[0059] In another embodiment, the methods of the present invention
may be used in combination with or to prevent or treat secondary
effects of: intraocular lens replacement; ophthalmic enucleation,
evisceration, exenteration, or a combination thereof; lacrimal sac
surgeries; corneal pterygium; lamellar keratoplasty; penetrating
keratoplasty, or a combination thereof, as well as any of the
disorders or conditions mentioned herein.
[0060] In one embodiment, the disease or disorder of the eye
affects the anterior region of the eye, while in another
embodiment, it affects the posterior region of the eye, while in
another embodiment, it affects both the anterior and posterior
regions of the eye. In one embodiment, the anterior segment
includes the cornea, anterior chamber, iris and ciliary body
(anterior choroid), posterior chamber and crystalline lens and the
posterior segment includes the retina with optic nerve, choroid
(posterior choroid) and vitreous. In one embodiment, eye disorders
resulting from the pathologic conditions of structures in the
anterior segment of the eye are dry eye syndrome, keratitis or
corneal dystrophy, cataracts, and glaucoma. In one embodiment, the
disease or disorders of the posterior segment of the eye in general
are retinal or choroidal vascular diseases or hereditary diseases
such as Lebers congenital amaurosis.
[0061] In one embodiment, "treating" refers to both therapeutic
treatment and prophylactic or preventive measures, wherein the
object is to prevent or lessen the targeted pathologic condition or
disorder as described hereinabove. Thus, in one embodiment,
treating may include suppressing, inhibiting, preventing, treating,
or a combination thereof. Thus, in one embodiment, "treating"
refers, inter alia, to increasing time to sustained progression,
expediting remission, inducing remission, augmenting remission,
speeding recovery, increasing efficacy of or decreasing resistance
to alternative therapeutics, or a combination thereof. In one
embodiment, "preventing" refers, inter alia, to delaying the onset
of symptoms, preventing relapse to a disease, decreasing the number
or frequency of relapse episodes, increasing latency between
symptomatic episodes, or a combination thereof. In one embodiment,
"suppressing" or "inhibiting", refers, inter alia, to reducing the
severity of symptoms, reducing the severity of an acute episode,
reducing the number of symptoms, reducing the incidence of
disease-related symptoms, reducing the latency of symptoms,
ameliorating symptoms, reducing secondary symptoms, reducing
secondary infections, prolonging patient survival, or a combination
thereof.
[0062] In one embodiment, symptoms are primary, while in another
embodiment, symptoms are secondary. In one embodiment, "primary"
refers to a symptom that is a direct result of an eye disease,
while in one embodiment, "secondary" refers to a symptom that is
derived from or consequent to a primary cause. In one embodiment,
the compounds for use in the present invention treat primary or
secondary symptoms or secondary complications related to an eye
disease. In another embodiment, the compounds for use in the
present invention treat primary or secondary symptoms or secondary
complications related to an eye disease or disorder.
[0063] In another embodiment, "symptoms" may be any manifestation
of a disease or pathological condition, comprising inflammation,
swelling, fever, pain, bleeding, itching, runny nose, coughing,
headache, migraine, dizziness, blurry vision, decreased visual
acuity, light sensitivity, etc., or a combination thereof. In one
embodiment, symptoms comprise itchy eyes, swollen eyelids, redness,
irritation, watery eyes, mucoid discharge, pain, or a combination
thereof.
[0064] Thus, in one embodiment of the present invention, the
compounds for use in the present invention are directed towards the
resolution of symptoms of a disease or disorder of the eye. In
another embodiment, the compounds affect the pathogenesis
underlying a disease or disorder of the eye.
[0065] In one embodiment, a disease or disorder of the eye may
affect a cell, in one embodiment, a vertebrate cell, in another
embodiment, a mammalian cell, and in another embodiment, a human
cell. It is to be understood that compounds of the present
invention may be efficacious in treating any cell type in which a
disease or disorder of the eye or the causes of a disease or
disorder of the eye may exert an effect. In one embodiment, a
compound for use in the present invention may localize to or act on
a specific cell type. In one embodiment, a compound for use in the
present invention may be cytoprotective. In one embodiment a
compound for use in the present invention may be inserted or
partially inserted into a cell membrane. In another embodiment a
compound for use in the present invention may be effective in
treating a plurality of cell types.
[0066] In one embodiment, a disease or disorder of the eye is a
primary or secondary symptom of an underlying illness, which in one
embodiment, is an autoimmune disease. In one embodiment, the
underlying illness is rheumatoid arthritis, systemic lupus
erythematosus, Kawasaki's Disease, ulcerative colitis, Crohn's
Disease, ankylosing spondylitis, Behcet's syndrome, psoriasis,
Reiter's syndrome, sarcoidosis, diabetes, multiple sclerosis, etc.,
or any combination thereof.
[0067] In one embodiment, the methods of the present invention may
be used to treat a disease or disorder of the eye in a subject that
is immunosuppressed, while in another embodiment, in a subject that
is immunodeficient, while in another embodiment, in a subject that
is immunocompetent.
[0068] In another embodiment, the methods of the present invention
may be used to prevent or treat glaucoma. In one embodiment,
glaucoma is characterized by increased fluid pressure in the eye,
which in one embodiment, is due to slowed fluid drainage from the
eye. In one embodiment, glaucoma may damage the optic nerve and
other parts of the eye, lead to vision loss or blindness, or a
combination thereof. In one embodiment, glaucoma may refer to
primary open angle glaucoma, normal pressure glaucoma, normal
tension glaucoma, pigmentary glaucoma, pseudoexfoliation glaucoma,
acute angle closure glaucoma, absolute glaucoma chronic glaucoma,
congenital glaucoma, juvenile glaucoma, narrow angle glaucoma,
chronic open angle glaucoma, simplex glaucoma, primary congenital
glaucoma, secondary glaucoma, or a combination thereof.
[0069] In another embodiment, the methods of the present invention
may be used to prevent or treat macular degeneration. In one
embodiment, macular degeneration is characterized by damage to or
breakdown of the macula, which in one embodiment, is a small area
at the back of the eye. In one embodiment, macular degeneration
causes a progressive loss of central sight, but not complete
blindness. In one embodiment, macular degeneration is of the dry
type, while in another embodiment, it is of the wet type. In one
embodiment, the dry type is characterized by the thinning and loss
of function of the macula tissue. In one embodiment, the wet type
is characterized by the growth of abnormal blood vessels behind the
macula. In one embodiment, the abnormal blood vessels hemorrhage or
leak, resulting in the formation of scar tissue if untreated. In
some embodiments, the dry type of macular degeneration can turn
into the wet type. In one embodiment, macular degeneration is
age-related, which in one embodiment is caused by an ingrowth of
chorioidal capillaries through defects in Bruch's membrane with
proliferation of fibrovascular tissue beneath the retinal pigment
epithelium.
[0070] In another embodiment, the methods of the present invention
may be used to prevent or treat retinopathy. In one embodiment,
retinopathy refers to a disease of the retina, which in one
embodiment is characterized by inflammation and in another
embodiment, is due to blood vessel damage inside the eye. In one
embodiment, retinopathy is diabetic retinopathy which, in one
embodiment, is a complication of diabetes that is caused by changes
in the blood vessels of the retina. In one embodiment, blood
vessels in the retina leak blood and/or grow fragile, brush-like
branches and scar tissue, which in one embodiment, blurs or
distorts the images that the retina sends to the brain. In another
embodiment, retinopathy is proliferative retinopathy, which in one
embodiment, is characterized by the growth of new, abnormal blood
vessels on the surface of the retina (neovascularization). In one
embodiment, neovascularization around the pupil increases pressure
within the eye, which in one embodiment, leads to glaucoma. In
another embodiment, neovascularization leads to new blood vessels
with weaker walls that break and bleed, or cause scar tissue to
grow, which in one embodiment, pulls the retina away from the back
of the eye (retinal detachment). In one embodiment, the
pathogenesis of retinopathy is related to non-enzymatic glycation,
glycoxidation, accumulation of advanced glycation end-products,
free radical-mediated protein damage, up-regulation of matrix
metalloproteinases, elaboration of growth factors, secretion of
adhesion molecules in the vascular endothelium, or a combination
thereof.
[0071] In one embodiment, retinopathy leads to macular edema, which
in one embodiment, is swelling of the retina. In one embodiment,
macular edema is characterized by retinal blood vessels that
develop tiny leaks, which in one embodiment, allow blood and fluid
to seep from the retinal blood vessels, and fatty material (called
exudate) to deposit in the retina. In one embodiment, symptoms of
macular edema comprise impaired or blurred vision.
[0072] In another embodiment, retinopathy refers to retinopathy of
prematurity (ROP), which in one embodiment, occurs in premature
babies when abnormal blood vessels and scar tissue grow over the
retina. In one embodiment, retinopathy of prematurity is caused by
a therapy necessary to promote the survival of a premature
infant.
[0073] In another embodiment, retinopathy refers to
arteriosclerotic retinopathy, which in one embodiment, is due to
arteriosclerosis (hardening of the arteries). In another
embodiment, retinopathy refers to hypertensive retinopathy, which
in one embodiment, is due to high blood pressure. In another
embodiment, retinopathy refers to solar retinopathy, while in
another embodiment, it refers to drug-related retinopathy.
[0074] In another embodiment, the methods of the present invention
may be used to prevent or treat retinal detachment, including,
inter alia, rhegmatogenous, tractional, or exudative retinal
detachment, which in one embodiment, is the separation of the
retina from its supporting layers. In one embodiment, retinal
detachment is associated with a tear or hole in the retina through
which the internal fluids of the eye may leak. In one embodiment,
retinal detachment is caused by trauma, the aging process, severe
diabetes, an inflammatory disorder, neovascularization, or
retinopathy of prematurity, while in another embodiment, it occurs
spontaneously. In one embodiment, bleeding from small retinal blood
vessels may cloud the vitreous during a detachment, which in one
embodiment, may cause blurred and distorted images. In one
embodiment, a retinal detachment can cause severe vision loss,
including blindness.
[0075] Administration of the compounds for use in the present
invention in a diversity of animal and cell models of disease
invoke remarkable, and unexpected, cytoprotective effects, which
are useful in the prevention and treatment of eye diseases and/or
conditions.
[0076] In one embodiment of the present invention, the useful
pharmacological properties of the compounds for use in the present
invention, some of which are described hereinabove, may be applied
for clinical use, and disclosed herein as methods for the
prevention or treatment of a disease. The biological basis of these
methods may be readily demonstrated by standard cellular and animal
models of disease, for example, as described in the Examples
hereinbelow.
[0077] In one embodiment, the pharmacological activities of
compounds for use in the present invention, including membrane
stabilization, anti-inflammation, anti-oxidant action, and
attenuation of chemokine levels, may contribute to the resistance
of a treated cell to diseases of the eye. In one embodiment, cell
membrane stabilization may ameliorate or prevent tissue injury
arising in the course of an eye disease. In another embodiment,
anti-oxidant action may limit oxidative damage to cell and blood
components arising in the course of an eye disease. In another
embodiment, attenuation of chemokine levels may attenuate
physiological reactions to stress that arise in the course of an
eye disease.
[0078] In one embodiment of the invention, the compounds for use in
the present invention described herein can be used to treat
disease, through amelioration or prevention, of tissue injury
arising in the course of pathological disease states by stabilizing
cell membranes; limiting oxidative damage to cell and blood
components; or attenuating physiological reactions to stress, as
expressed in elevated chemokine levels.
[0079] In one embodiment, methods of the present invention involve
treating a subject by, inter alia, controlling the expression,
production, and activity of phospholipases such as PLA2;
controlling the production and/or action of lipid mediators, such
as eicosanoids, platelet activating factor (PAF) and
lyso-phospholipids; amelioration of damage to cell surface
glycosaminoglycans (GAG) and proteoglycans; controlling the
production of oxidants, oxygen radicals and nitric oxide;
protection of cells, tissues, and plasma lipoproteins from damaging
agents, such as reactive oxygen species (ROS) and phospholipases;
controlling the expression, production, and activity of cytokines,
chemokines and interleukins; anti-oxidant therapy; anti-endotoxin
therapy or any combination thereof.
[0080] In one embodiment of the invention, the term "controlling"
refers to inhibiting the production and action of the above
mentioned factors in order to maintain their activity at the normal
basal level and suppress their activation in pathological
conditions.
[0081] In one embodiment of the invention, eye disease is
characterized by the presence of damaging agents, which comprise,
inter alia, phospholipases, reactive oxygen species (ROS), free
radicals, lysophospholipids, fatty acids or derivatives thereof,
hydrogen peroxides, phospholipids, oxidants, cationic proteins,
streptolysins, proteases, hemolysins, or sialidases.
Dosages and Routes of Administration
[0082] This invention encompasses administration of compounds as
described herein or compositions comprising the same, for treating
diseases of the eye.
[0083] In one embodiment, compositions of this invention are
pharmaceutically acceptable. In one embodiment, the term
"pharmaceutically acceptable" refers to any formulation which is
safe, and provides the appropriate delivery for the desired route
of administration of an effective amount of at least one compound
for use in the present invention. This term refers to the use of
buffered formulations as well, wherein the pH is maintained at a
particular desired value, ranging from pH 4.0 to pH 9.0, in
accordance with the stability of the compounds and route of
administration.
[0084] In some embodiments, any of the compositions of this
invention will comprise a lipid conjugate, in any form or
embodiment as described herein. In some embodiments, any of the
compositions of this invention will consist of a lipid conjugate,
in any form or embodiment as described herein. In some embodiments,
of the compositions of this invention will consist essentially of a
lipid conjugate, in any form or embodiment as described herein. In
some embodiments, the term "comprise" refers to the inclusion of
the indicated active agent, such as the Compounds I-C, as well as
inclusion of other active agents, and pharmaceutically acceptable
carriers, excipients, emollients, stabilizers, etc., as are known
in the pharmaceutical industry. In some embodiments, the term
"consisting essentially of" refers to a composition, whose only
active ingredient is the indicated active ingredient, however,
other compounds may be included which are for stabilizing,
preserving, etc. the formulation, but are not involved directly in
the therapeutic effect of the indicated active ingredient. In some
embodiments, the term "consisting essentially of" may refer to
components which facilitate the release of the active ingredient.
In some embodiments, the term "consisting" refers to a composition,
which contains the active ingredient and a pharmaceutically
acceptable carrier or excipient.
[0085] In one embodiment, a compound used in the methods of this
invention may be administered alone or within a composition. In
another embodiment, compositions comprising compounds for use in
the present invention in admixture with conventional excipients,
i.e. pharmaceutically acceptable organic or inorganic carrier
substances suitable for parenteral, enteral (e.g. oral) or topical
application which do not deleteriously react with the active
compounds may be used. In one embodiment, suitable pharmaceutically
acceptable carriers include but are not limited to water, salt
solutions, alcohols, gum arabic, vegetable oils, benzyl alcohols,
polyethylene glycols, gelatine, carbohydrates such as lactose,
amylose or starch, magnesium stearate, talc, silicic acid, viscous
paraffin, white paraffin, glycerol, alginates, hyaluronic acid,
collagen, perfume oil, fatty acid monoglycerides and diglycerides,
pentaerythritol fatty acid esters, hydroxy methylcellulose,
polyvinyl pyrrolidone, etc. In another embodiment, the
pharmaceutical preparations can be sterilized and if desired mixed
with auxiliary agents, e.g. lubricants, preservatives, stabilizers,
wetting agents, emulsifiers, salts for influencing osmotic
pressure, buffers, coloring, flavoring and/or aromatic substances
and the like which do not deleteriously react with the active
compounds. In another embodiment, they can also be combined where
desired with other active agents, e.g. vitamins.
[0086] In one embodiment, the therapeutic compositions of the
instant invention comprise a compound of the instant invention and
additional compounds effective in preventing or treating eye
disease. In one embodiment, the additional compounds comprise
anti-inflammatory compositions, which in one embodiment are
non-steroidal anti-inflammatory medications, antihistamines,
antibiotics, corticosteroids, cromolyn sodium (sodium
cromoglicate), mast-cell stabilizers, artificial tears, lubricants,
or a combination thereof. In one embodiment, antibiotics comprise
chloramphenicol, fusidic acid, tetracycline, erythromycin,
gentamycin, or a combination thereof. In another embodiment, an
additional compound is vitamin A.
[0087] In one embodiment, the therapeutic compositions of the
instant invention are administered with other treatments that
relieve symptoms. In one embodiment, other treatments comprise
application of cold compresses, while in another embodiment, warm
compresses.
[0088] In one embodiment, the route of administration may be
parenteral, enteral, or a combination thereof. In another
embodiment, the route may be intra-ocular, topical, transdermal,
intradermal, subcutaneous, intraperitoneal, intravenous,
intra-arterial, vaginal, rectal, intratumoral, parcanceral,
transmucosal, intramuscular, intravascular, intraventricular,
intracranial, inhalation, nasal aspiration (spray), sublingual,
oral, aerosol or suppository or a combination thereof. In one
embodiment, the dosage regimen will be determined by skilled
clinicians, based on factors such as exact nature of the condition
being treated, the severity of the condition, the age and general
physical condition of the patient, etc.
[0089] For intra-ocular application, eye drops, ointments, lotions,
creams, or coated eye patches may be used in one embodiment. In
another embodiment, intra-ocular application may comprise the use
of contact lens comprising the compounds of the instant
invention.
[0090] In one embodiment, intra-ocular application is used to treat
an eye condition or disease. In another embodiment, intra-ocular
injection is used to treat an eye condition or disease. In one
embodiment, compounds may be administered intravitreally, in
another embodiment, subretinally, while in another embodiment,
intra-retinally, while in another embodiment, periocularly. In one
embodiment, compounds may be administered intracamerally into the
anterior chamber or vitreous, via a depot attached to the
intraocular lens implant inserted during surgery, or via a depot
placed in the eye sutured in the anterior chamber or vitreous.
[0091] For parenteral application, particularly suitable are
injectable, sterile solutions, preferably oily or aqueous
solutions, as well as suspensions, emulsions, or implants,
including suppositories and enemas. Ampoules are convenient unit
dosages. Such a suppository may comprise any agent described
herein.
[0092] For application by inhalation, solutions or suspensions of
the compounds mixed and aerosolized or nebulized in the presence of
the appropriate carrier suitable. Such an aerosol may comprise any
agent described herein and, in one embodiment, may be used to treat
diseases or conditions caused by airborne pathogens, which may in
one embodiment, cause sinusitis or upper respiratory infections, in
addition to eye diseases.
[0093] For topical application, particularly in the area around the
eye, an admixture of the compounds with conventional creams,
lotions, or delayed release patches is acceptable. Such a cream or
lotion may comprise any agent described herein, and, in one
embodiment, may be used to treat an eye disease.
[0094] For enteral application, particularly suitable are tablets,
dragees, liquids, drops, or capsules. A syrup, elixir, or the like
can be used when a sweetened vehicle is employed.
[0095] Sustained or directed release compositions can be
formulated, e.g. liposomes or those wherein the active compound is
protected with differentially degradable coatings, e.g. by
microencapsulation, multiple coatings, etc. It is also possible to
freeze-dry the new compounds and use the lyophilisates obtained,
for example, for the preparation of products for injection.
[0096] Thus, in one embodiment, the route of administration may be
directed to an organ or system that is affected by an eye disease.
For example, compounds may be administered in intra-ocular form to
treat an eye disease. In another embodiment, the route of
administration may be directed to a different organ or system than
the one that is affected by an eye disease. For example, compounds
may be administered parenterally to treat an eye disease. Thus, the
present invention provides for the use of compounds of the instant
invention in various dosage forms suitable for administration using
any of the routes listed hereinabove.
[0097] In general, the doses utilized for the above described
purposes will vary, but will be in an effective amount to exert the
desired effect. As used herein, the term "pharmaceutically
effective amount" refers to an amount of a compound of formulae A
and I-LXXXVII as described hereinbelow, which will produce the
desired alleviation in symptoms or other desired phenotype in a
patient. The doses utilized for any of the above-described purposes
will generally be from 1 to about 1000 milligrams per kilogram of
body weight (mg/kg), administered one to four times per day, or by
continuous IV infusion. When the compositions are dosed topically
or intraocularly, they will generally be in a concentration range
of from 0.1 to about 10% w/v, administered 1-4 times per day.
[0098] In one embodiment of the invention, the concentrations of
the compounds will depend on various factors, including the nature
of the condition to be treated, the condition of the patient, the
route of administration and the individual tolerability of the
compositions.
[0099] It will be appreciated that the actual preferred amounts of
active compound in a specific case will vary according to the
specific compound being utilized, the particular compositions
formulated, the mode of application, and the particular conditions
and organism being treated. Dosages for a given host can be
determined using conventional considerations, e.g. by customary
comparison of the differential activities of the subject compounds
and of a known agent, e.g. by means of an appropriate, conventional
pharmacological protocol.
[0100] In one embodiment, the compounds of the invention may be
administered acutely for acute treatment of temporary conditions,
or may be administered chronically, especially in the case of
progressive, recurrent, or degenerative disease. In one embodiment,
one or more compounds of the invention may be administered
simultaneously, or in another embodiment, they may be administered
in a staggered fashion. In one embodiment, the staggered fashion
may be dictated by the stage or phase of the disease.
[0101] In one embodiment, the present invention offers methods for
the treatment of disease based upon administration of lipids
covalently conjugated through their polar head group to a
physiologically acceptable chemical moiety, which may be of high or
low molecular weight.
[0102] The present invention has been illustrated in terms of the
anti-disease activity of compounds for use in the present invention
and methods of their use as pharmaceutical compositions in the
treatment of disease. The following sections present some examples
of the therapeutic compounds for use in the present invention and
their chemical preparation.
Compounds
[0103] In one embodiment, the compounds for use in the present
invention or for the compositions of the present invention comprise
a lipid or phospholipid moiety bound to a physiologically
acceptable monomer, dimer, oligomer, or polymer. In one embodiment,
the physiologically acceptable monomer, dimer, oligomer, or polymer
is salicylate, salicylic acid, aspirin, a monosaccharide,
lactobionic acid, maltose, an amino acid, glycine, carboxylic acid,
acetic acid, butyric acid, dicarboxylic acid, glutaric acid,
succinic acid, fatty acid, dodecanoic acid, didodecanoic acid, bile
acid, cholic acid, cholesterylhemmisuccinate, a dipeptide, a
disaccharide, a trisaccharide, an oligosaccharide, a
polysaccharide, a hetero-polysaccharide, a homo-polysaccharide, a
polypyranose, an oligopeptide, or a di- or trisaccharide monomer
unit of heparin, heparan sulfate, keratin, keratan sulfate,
chondroitin, chondroitin-6-sulfate, chondroitin-4-sulfate,
dermatin, dermatan sulfate, dextran, or hyaluronic acid, a
glycosaminoglycan, polygeline (`haemaccel`), alginate, hydroxyethyl
starch (hetastarch), polyethylene glycol, polycarboxylated
polyethylene glycol, chondroitin-6-sulfate, chondroitin-4-sulfate,
keratin, keratin sulfate, heparan sulfate, dermatin, dermatan
sulfate, carboxymethylcellulose, heparin, dextran, or hyaluronic
acid.
[0104] In one embodiment, examples of polymers which can be
employed as the conjugated moiety for producing compounds for use
in the present invention for use in the methods of this invention
may be physiologically acceptable polymers, including
water-dispersible or -soluble polymers of various molecular weights
and diverse chemical types, mainly natural and synthetic polymers,
such as glycosaminoglycans, hyaluronic acids, heparin, heparin
sulfates, chondroitin sulfates, chondroitin-6-sulfates,
chondroitin-4-sulfates, keratins, keratin sulfates, dermatins,
dermatan sulfates, dextrans, plasma expanders, including polygeline
("Haemaccel", degraded gelatin polypeptide cross-linked via urea
bridges, produced by "Behring"), "hydroxyethylstarch" (Hetastarch,
HES) and extrans, food and drug additives, soluble cellulose
derivatives (e.g. methylcellulose, carboxymethylcellulose),
polyaminoacids, hydrocarbon polymers (e.g. polyethylene),
polystyrenes, polyesters, polyamides, polyethylene oxides (e.g.
polyethyleneglycols, polycarboxyethyleneglycols, polycarboxylated
polyethyleneglycols), polyvinnylpyrrolidones, polysaccharides,
polypyranoses, alginates, assimilable gums (e.g. xanthan gum),
peptides, injectable blood proteins (e.g. serum albumin),
cyclodextrin, and derivatives thereof.
[0105] In one embodiment, examples of monomers, dimers, and
oligomers which can be employed as the conjugated moiety for
producing compounds for use in the present invention for use in the
methods of the invention may be mono- or disaccharides,
trisaccharides, oligopeptides, carboxylic acids, dicarboxylic
acids, fatty acids, dicarboxylic fatty acids, salicylates,
slicyclic acids, acetyl salicylic acids, aspirins, lactobionic
acids, maltoses, amino acids, glycines, glutaric acids, succinic
acids, dodecanoic acids, didodecanoic acids, bile acids, cholic
acids, cholesterylhemisuccinates, and di- and trisaccharide unit
monomers of polysaccharides, polypyranoses, and/or
glycosaminoglycans including heparins, heparan sulfates, hyaluronic
acids, chondroitins, chondroitin sulfates, chondroitin-6-sulfates,
chondroitin-4-sulfates, dermatins, dermatan sulfates, keratins,
keratan sulfates, or dextrans.
[0106] In one embodiment, the lipid compounds for use in the
present invention are described by the general formula:
[phosphatidylethanolamine-Y]n-X
[phosphatidylserine-Y]n-X
[phosphatidylcholine-Y]n-X
[phosphatidylinositol-Y]n-X
[phosphatidylglycerol-Y]n-X
[phosphatidic acid-Y]n-X
[lyso-phospholipid-Y]n-X
[diacyl-glycerol-Y]n-X
[monoacyl-glycerol-Y]n-X
[sphingomyelin-Y]n-X
[sphingosine-Y]n-X
[ceramide-Y]n-X
[0107] wherein [0108] Y is either nothing or a spacer group ranging
in length from 2 to 30 atoms; and [0109] X is a physiologically
acceptable monomer, dimer, oligomer or polymer; and [0110] n is the
number of lipid molecules bound to a molecule of X, wherein n is a
number from 1 to 1000. In another embodiment, n is a number from 2
to 1000.
[0111] In one embodiment, the invention provides low-molecular
weight compounds, previously undisclosed and unknown to possess
pharmacological activity, of the general formula described
hereinabove. In another embodiment, wherein the general formula
described hereinabove describes low-molecular weight compounds, X
is a mono- or disaccharide, carboxylated disaccharide, mono- or
dicarboxylic acids, a salicylate, salicylic acid, aspirin,
lactobionic acid, maltose, an amino acid, glycine, acetic acid,
butyric acid, dicarboxylic acid, glutaric acid, succinic acid,
fatty acid, dodecanoic acid, didodecanoic acid, bile acid, cholic
acid, cholesterylhemmisuccinate, a di- or tripeptide, an
oligopeptide, a trisacharide, or a di- or trisaccharide monomer
unit of heparin, heparan sulfate, keratin, keratan sulfate,
chondroitin, chondroitin-6-sulfate, chondroitin-4-sulfate,
dermatin, dermatan sulfate, dextran, or hyaluronic acid.
[0112] In one embodiment of this invention, X is any of the
physiologically acceptable monomer, dimer, oligomer, or polymer, as
described herein. In one embodiment, X is conjugated to the lipid,
phospholipid, or spacer via an ester bond. In another embodiment, X
is conjugated to the lipid, phospholipid, or spacer via an amide
bond.
[0113] As defined by the structural formulae provided herein for
the compounds for use in the present invention, these compounds may
contain between one to one thousand lipid moieties bound to a
single physiologically acceptable polymer molecule. In one
embodiment of this invention, n is a number from 1 to 1000. In
another embodiment, n is a number from 1 to 500. In another
embodiment, n is a number from 1 to 100. In another embodiment, n
is a number from 1 to 50. In another embodiment, n is a number from
1 to 25. In another embodiment, n is a number from 1 to 10. In
another embodiment, n is a number from 1-5. In another embodiment,
n is a number from 1 to 4. In another embodiment, n is a number
from 1 to 3, In another embodiment, n is a number from 1 to 2. In
another embodiment, n is a number from 2 to 1000. In another
embodiment, n is a number from 2 to 100. In another embodiment, n
is a number from 2 to 200. In another embodiment, n is a number
from 2 to 50. In another embodiment, n is a number from 2 to 25. In
another embodiment, n is a number from 2-10. In another embodiment,
n is a number from 2 to 5. In another embodiment, n is a number
from 2 to 4, In another embodiment, n is a number from 2 to 3. In
another embodiment, n is a number from 3 to 300. In another
embodiment, n is a number from 10 to 400. In another embodiment, n
is a number from 50 to 500. In another embodiment, n is a number
from 100 to 300. In another embodiment, n is a number from 300 to
500. In another embodiment, n is a number from 500 to 800. In
another embodiment, n is a number from 500 to 1000.
[0114] In one embodiment of the invention, when the conjugated
moiety is a polymer, the ratio of lipid moieties covalently bound
may range from one to one thousand lipid residues per polymer
molecule, depending upon the nature of the polymer and the reaction
conditions employed. For example, the relative quantities of the
starting materials, or the extent of the reaction time, may be
modified in order to obtain products with either high or low ratios
of lipid residues per polymer, as desired.
[0115] In one embodiment, the set of compounds comprising
phosphatidylethanolamine covalently bound to a physiologically
acceptable monomer, dimmer, oligomer, or polymer, is referred to
herein as the PE-conjugates. In one embodiment, the
phosphatidylethanolamine moiety is dipalmitoyl
phosphatidylethanolamine. In another embodiment, the
phosphatidylethanolamine moiety is dimyristoyl
phosphatidylethanolamine. In another embodiment, related
derivatives, in which either phosphatidylserine,
phosphatidylcholine, phosphatidylinositol, phosphatidic acid or
phosphatidylglycerol are employed in lieu of
phosphatidylethanolamine as the lipid moiety provide equivalent
therapeutic results, based upon the biological experiments
described below for the compounds for use in the present invention
and the structural similarities shared by these compounds.
[0116] In another embodiment, the lipid or phospholipid moiety is
phosphatidic acid, an acyl glycerol, monoacylglycerol,
diacylglycerol, triacylglycerol, sphingosine, sphingomyelin,
chondroitin-4-sulfate, chondroitin-6-sulfate, ceramide,
phosphatidylethanolamine, phosphatidylserine, phosphatidylcholine,
phosphatidylinositol, or phosphatidylglycerol, or an ether or alkyl
phospholipid derivative thereof.
[0117] In one embodiment, derivatives relevant to this invention
are compounds wherein at least one of the fatty acid groups of the
lipid moieties at position C1 or C2 of the glycerol backbone are
substituted by a long chain alkyl group attached by amide, ether or
alkyl bonds, rather than ester linkages.
[0118] In the methods, according to embodiments of the invention,
the compounds for use in the present invention administered to the
subject are comprised from at least one lipid moiety covalently
bound through an atom of the polar head group to a monomeric or
polymeric moiety (referred to herein as the conjugated moiety) of
either low or high molecular weight. When desired, an optional
bridging moiety can be used to link the compounds for use in the
present invention moiety to the monomer or polymeric moiety. The
conjugated moiety may be a low molecular weight carboxylic acid,
dicarboxylic acid, fatty acid, dicarboxylic fatty acid, acetyl
salicylic acid, cholic acid, cholesterylhemisuccinate, or mono- or
di-saccharide, an amino acid or dipeptide, an oligopeptide, a
glycoprotein mixture, a di- or trisaccharide monomer unit of a
glycosaminoglycan such as a repeating unit of heparin, heparan
sulfate, hyaluronic acid, chondroitin-sulfate, dermatan, keratan
sulfate, or a higher molecular weight peptide or oligopeptide, a
polysaccharide, a hetero-polysaccharide, a homo-polysaccharide, a
polypyranose, polyglycan, protein, glycosaminoglycan, or a
glycoprotein mixture. The composition of some
phospholipid-conjugates of high molecular weight, and associated
analogues, are the subject of U.S. Pat. No. 5,064,817, which is
incorporated herein in its entirety by reference.
[0119] In one embodiment, the term "moiety" means a chemical entity
otherwise corresponding to a chemical compound, which has a valence
satisfied by a covalent bond.
[0120] In some cases, according to embodiments of the invention,
the monomer or polymer chosen for preparation of the compound may
in itself have select biological properties. For example, both
heparin and hyaluronic acid are materials with known physiological
functions. In the present invention, however, the compounds for use
in the present invention formed from these substances as starting
materials display a new and wider set of pharmaceutical activities
than would be predicted from administration of either heparin or
hyaluronic acid which have not been bound by covalent linkage to a
phospholipid. In some embodiments, phosphatidylethanolamine (PE)
linked to hyaluronic acid (Compound XXII), to heparin (Compound
XXIV), to chondroitin sulfate A (Compound XXV), to
carboxymethylcellulose (Compound XXVI), to Polygeline (haemaccel)
(Compound XXVII), to alginate (Compound LI), or to
hydroxyethylstarch (Compound XXVIII), are useful for methods and in
compositions as herein described but perform unexpectedly in terms
of potency and range of useful pharmaceutical activity compared to
the free conjugates. Thus, the combination of a phospholipid such
as phosphatidylethanolamine, or related phospholipids which differ
with regard to the polar head group, such as phosphatidylserine
(PS), phosphatidylcholine (PC), phosphatidylinositol (PI), and
phosphatidylglycerol (PG), results in the formation of a compound
which has novel pharmacological properties when compared to the
starting materials alone. In one embodiment, such properties may
include: greater lubrication, greater local persistence, greater
anti-inflammatory properties, greater antioxidant activity, or a
combination thereof.
[0121] The biologically active compounds for use in the present
invention described herein can have a wide range of molecular
weights, e.g. above 50,000 (up to a few hundred thousands) when it
is desirable to retain the lipid conjugate in the vascular system
and below 50,000 when targeting to extravascular systems is
desirable. The sole limitation on the molecular weight and the
chemical structure of the conjugated moiety is that it does not
result in a compound devoid of the desired biological activity, or
lead to chemical or physiological instability to the extent that
the Compound is rendered useless as a drug in the method of use
described herein.
[0122] In one embodiment, the compound for use in the present
invention is represented by the structure of the general formula
(A):
##STR00005##
[0123] wherein [0124] L is a lipid or a phospholipid; [0125] Z is
either nothing, ethanolamine, serine, inositol, choline, phosphate,
or glycerol; [0126] Y is either nothing or a spacer group ranging
in length from 2 to 30 atoms; [0127] X is a physiologically
acceptable monomer, dimer, oligomer, or polymer; and [0128] n is a
number from 1 to 1000; [0129] wherein any bond between L, Z, Y and
X is either an amide or an esteric bond.
[0130] In one embodiment, the compound for use in the present
invention is represented by the structure of the general formula
(A):
##STR00006##
[0131] wherein [0132] L is a lipid or a phospholipid; [0133] Z is
either nothing, ethanolamine, serine, inositol, choline, phosphate,
or glycerol; [0134] Y is either nothing or a spacer group ranging
in length from 2 to 30 atoms; [0135] X is a physiologically
acceptable monomer, dimer, oligomer, or polymer; and [0136] n is a
number from 2 to 1000; [0137] wherein any bond between L, Z, Y and
X is either an amide or an esteric bond.
[0138] In one embodiment, L is phosphatidyl, Z is ethanolamine,
wherein L and Z are chemically bonded resulting in
phosphatidylethanolamine, Y is nothing, and X is
carboxymethylcellulose. In another embodiment, L is phosphatidyl, Z
is ethanolamine, wherein L and Z are chemically bonded resulting in
phosphatidylethanolamine, Y is nothing, and X is a
glycosaminoglycan. In one embodiment, the phosphatidylethanolamine
moiety is dipalmitoyl phosphatidylethanolamine. In another
embodiment, the phosphatidylethanolamine moiety is dimyristoyl
phosphatidylethanolamine.
[0139] In another embodiment, the compound for use in the present
invention is represented by the structure of the general formula
(I):
##STR00007##
[0140] wherein [0141] R.sub.1 is a linear, saturated,
mono-unsaturated, or poly-unsaturated, alkyl chain ranging in
length from 2 to 30 carbon atoms; [0142] R.sub.2 is a linear,
saturated, mono-unsaturated, or poly-unsaturated, alkyl chain
ranging in length from 2 to 30 carbon atoms; [0143] Y is either
nothing or a spacer group ranging in length from 2 to 30 atoms; and
[0144] X is either a physiologically acceptable monomer, dimer,
oligomer or a physiologically acceptable polymer; and [0145] n is a
number from 1 to 1,000; [0146] wherein if Y is nothing the
phosphatidylethanolamine is directly linked to X via an amide bond
and if Y is a spacer, the spacer is directly linked to X via an
amide or an esteric bond and to the phosphatidylethanolamine via an
amide bond.
[0147] In another embodiment, the compound for use in the present
invention is represented by the structure of the general formula
(I):
##STR00008##
[0148] wherein [0149] R.sub.1 is a linear, saturated,
mono-unsaturated, or poly-unsaturated, alkyl chain ranging in
length from 2 to 30 carbon atoms; [0150] R.sub.2 is a linear,
saturated, mono-unsaturated, or poly-unsaturated, alkyl chain
ranging in length from 2 to 30 carbon atoms; [0151] Y is either
nothing or a spacer group ranging in length from 2 to 30 atoms; and
[0152] X is either a physiologically acceptable monomer, dimer,
oligomer or a physiologically acceptable polymer; and [0153] n is a
number from 2 to 1,000; [0154] wherein if Y is nothing the
phosphatidylethanolamine is directly linked to X via an amide bond
and if Y is a spacer, the spacer is directly linked to X via an
amide or an esteric bond and to the phosphatidylethanolamine via an
amide bond.
[0155] In one embodiment, compounds for use in the methods of the
invention comprise one of the following as the conjugated moiety X:
acetate, butyrate, glutarate, succinate, dodecanoate,
didodecanoate, maltose, lactobionic acid, dextran, alginate,
aspirin, cholate, cholesterylhemisuccinate,
carboxymethyl-cellulose, heparin, hyaluronic acid, chondroitin
sulfate, polygeline (haemaccel), hydroxyethylstarch (Hetastarch,
HES) polyethyleneglycol, polycarboxylated polyethylene glycol, a
glycosaminoglycan, a polysaccharide, a hetero-polysaccharide, a
homo-polysaccharide, or a polypyranose. The polymers used as
starting material to prepare the PE-conjugates may vary in
molecular weight from 1 to 2,000 kDa.
[0156] Examples of phosphatidylethanolamine (PE) moieties are
analogues of the phospholipid in which the chain length of the two
fatty acid groups attached to the glycerol backbone of the
phospholipid varies from 2-30 carbon atoms length, and in which
these fatty acids chains contain saturated and/or unsaturated
carbon atoms. In lieu of fatty acid chains, alkyl chains attached
directly or via an ether linkage to the glycerol backbone of the
phospholipid are included as analogues of PE. In one embodiment,
the PE moiety is dipalmitoyl-phosphatidyl-ethanolamine. In another
embodiment, the PE moiety is
dimyristoyl-phosphatidyl-ethanolamine.
[0157] Phosphatidyl-ethanolamine and its analogues may be from
various sources, including natural, synthetic, and semi-synthetic
derivatives and their isomers.
[0158] Phospholipids which can be employed in lieu of the PE moiety
are N-methyl-PE derivatives and their analogues, linked through the
amino group of the N-methyl-PE by a covalent bond; N,N-dimethyl-PE
derivatives and their analogues linked through the amino group of
the N,N-dimethyl-PE by a covalent bond, phosphatidylserine (PS) and
its analogues, such as palmitoyl-stearoyl-PS, natural PS from
various sources, semi-synthetic PSs, synthetic, natural and
artifactual PSs and their isomers. Other phospholipids useful as
conjugated moieties in this invention are phosphatidylcholine (PC),
phosphatidylinositol (PI), phosphatidic acid and
phosphoatidylglycerol (PG), as well as derivatives thereof
comprising either phospholipids, lysophospholipids, phosphatidic
acid, sphingomyelins, lysosphingomyelins, ceramide, and
sphingosine.
[0159] For PE-conjugates and PS-conjugates, the phospholipid is
linked to the conjugated monomer or polymer moiety through the
nitrogen atom of the phospholipid polar head group, either directly
or via a spacer group. For PC, PI, and PG conjugates, the
phospholipid is linked to the conjugated monomer or polymer moiety
through either the nitrogen or one of the oxygen atoms of the polar
head group, either directly or via a spacer group.
[0160] In another embodiment, the compound for use in the present
invention is represented by the structure of the general formula
(II):
##STR00009##
[0161] wherein [0162] R.sub.1 is a linear, saturated,
mono-unsaturated, or poly-unsaturated, alkyl chain ranging in
length from 2 to 30 carbon atoms; [0163] R.sub.2 is a linear,
saturated, mono-unsaturated, or poly-unsaturated, alkyl chain
ranging in length from 2 to 30 carbon atoms; [0164] Y is either
nothing or a spacer group ranging in length from 2 to 30 atoms;
[0165] X is a glycosaminoglycan; and [0166] n is a number from 1 to
1000; [0167] wherein if Y is nothing, the phosphatidylserine is
directly linked to X via an amide bond and if Y is a spacer, the
spacer is directly linked to X via an amide or an esteric bond and
to the phosphatidylserine via an amide bond.
[0168] In another embodiment, the compound for use in the present
invention is represented by the structure of the general formula
(II):
##STR00010##
[0169] wherein [0170] R.sub.1 is a linear, saturated,
mono-unsaturated, or poly-unsaturated, alkyl chain ranging in
length from 2 to 30 carbon atoms; [0171] R.sub.2 is a linear,
saturated, mono-unsaturated, or poly-unsaturated, alkyl chain
ranging in length from 2 to 30 carbon atoms; [0172] Y is either
nothing or a spacer group ranging in length from 2 to 30 atoms;
[0173] X is a glycosaminoglycan; and [0174] n is a number from 2 to
1000; [0175] wherein if Y is nothing, the phosphatidylserine is
directly linked to X via an amide bond and if Y is a spacer, the
spacer is directly linked to X via an amide or an esteric bond and
to the phosphatidylserine via an amide bond.
[0176] In one embodiment, the phosphatidylserine may be bonded to
Y, or to X if Y is nothing, via the COO.sup.- moiety of the
phosphatidylserine.
[0177] In another embodiment, the compound for use in the present
invention is represented by the structure of the general formula
(III):
##STR00011##
[0178] wherein [0179] R.sub.1 is a linear, saturated,
mono-unsaturated, or poly-unsaturated, alkyl chain ranging in
length from 2 to 30 carbon atoms; [0180] R.sub.2 is a linear,
saturated, mono-unsaturated, or poly-unsaturated, alkyl chain
ranging in length from 2 to 30 carbon atoms; [0181] Z is either
nothing, inositol, choline, or glycerol; [0182] Y is either nothing
or a spacer group ranging in length from 2 to 30 atoms; [0183] X is
a glycosaminoglycan; and [0184] n is a number from 1 to 1000;
[0185] wherein any bond between the phosphatidyl, Z, Y and X is
either an amide or an esteric bond.
[0186] In another embodiment, the compound for use in the present
invention is represented by the structure of the general formula
(IV):
##STR00012##
[0187] wherein [0188] R.sub.1 is either hydrogen or a linear,
saturated, mono-unsaturated, or poly-unsaturated, alkyl chain
ranging in length from 2 to 30 carbon atoms; [0189] R.sub.2 is a
linear, saturated, mono-unsaturated, or poly-unsaturated, alkyl
chain ranging in length from 2 to 30 carbon atoms; [0190] Z is
either nothing, inositol, choline, or glycerol; [0191] Y is either
nothing or a spacer group ranging in length from 2 to 30 atoms;
[0192] X is a glycosaminoglycan; and [0193] n is a number from 1 to
1000; [0194] wherein any bond between the phospholipid, Z, Y and X
is either an amide or an esteric bond.
[0195] In another embodiment, the compound for use in the present
invention is represented by the structure of the general formula
(III):
##STR00013##
[0196] wherein [0197] R.sub.1 is a linear, saturated,
mono-unsaturated, or poly-unsaturated, alkyl chain ranging in
length from 2 to 30 carbon atoms; [0198] R.sub.2 is a linear,
saturated, mono-unsaturated, or poly-unsaturated, alkyl chain
ranging in length from 2 to 30 carbon atoms; [0199] Z is either
nothing, inositol, choline, or glycerol; [0200] Y is either nothing
or a spacer group ranging in length from 2 to 30 atoms; [0201] X is
a glycosaminoglycan; and [0202] n is a number from 2 to 1000;
[0203] wherein any bond between the phosphatidyl, Z, Y and X is
either an amide or an esteric bond.
[0204] In another embodiment, the compound for use in the present
invention is represented by the structure of the general formula
(IV):
##STR00014##
[0205] wherein [0206] R.sub.1 is either hydrogen or a linear,
saturated, mono-unsaturated, or poly-unsaturated, alkyl chain
ranging in length from 2 to 30 carbon atoms; [0207] R.sub.2 is a
linear, saturated, mono-unsaturated, or poly-unsaturated, alkyl
chain ranging in length from 2 to 30 carbon atoms; [0208] Z is
either nothing, inositol, choline, or glycerol; [0209] Y is either
nothing or a spacer group ranging in length from 2 to 30 atoms;
[0210] X is a glycosaminoglycan; and [0211] n is a number from 1 to
1000; [0212] wherein any bond between the phospholipid, Z, Y and X
is either an amide or an esteric bond.
[0213] In another embodiment, the compound for use in the present
invention is represented by the structure of the general formula
(IV):
##STR00015##
[0214] wherein [0215] R.sub.1 is either hydrogen or a linear,
saturated, mono-unsaturated, or poly-unsaturated, alkyl chain
ranging in length from 2 to 30 carbon atoms; [0216] R.sub.2 is a
linear, saturated, mono-unsaturated, or poly-unsaturated, alkyl
chain ranging in length from 2 to 30 carbon atoms; [0217] Z is
either nothing, inositol, choline, or glycerol; [0218] Y is either
nothing or a spacer group ranging in length from 2 to 30 atoms;
[0219] X is a glycosaminoglycan; and [0220] n is a number from 2 to
1000; [0221] wherein any bond between the phospholipid, Z, Y and X
is either an amide or an esteric bond.
[0222] In another embodiment, the compound for use in the present
invention is represented by the structure of the general formula
(V):
##STR00016##
[0223] wherein [0224] R.sub.1 is a linear, saturated,
mono-unsaturated, or poly-unsaturated, alkyl chain ranging in
length from 2 to 30 carbon atoms; [0225] R.sub.2 is either hydrogen
or a linear, saturated, mono-unsaturated, or poly-unsaturated,
alkyl chain ranging in length from 2 to 30 carbon atoms; [0226] Z
is either nothing, inositol, choline, or glycerol; [0227] Y is
either nothing or a spacer group ranging in length from 2 to 30
atoms; [0228] X is a glycosaminoglycan; and [0229] n is a number
from 1 to 1000; [0230] wherein any bond between the phospholipid,
Z, Y and X is either an amide or an esteric bond.
[0231] In another embodiment, the compound for use in the present
invention is represented by the structure of the general formula
(V):
##STR00017##
[0232] wherein [0233] R.sub.1 is a linear, saturated,
mono-unsaturated, or poly-unsaturated, alkyl chain ranging in
length from 2 to 30 carbon atoms; [0234] R.sub.2 is either hydrogen
or a linear, saturated, mono-unsaturated, or poly-unsaturated,
alkyl chain ranging in length from 2 to 30 carbon atoms; [0235] Z
is either nothing, inositol, choline, or glycerol; [0236] Y is
either nothing or a spacer group ranging in length from 2 to 30
atoms; [0237] X is a glycosaminoglycan; and [0238] n is a number
from 2 to 1000; [0239] wherein any bond between the phospholipid,
Z, Y and X is either an amide or an esteric bond.
[0240] In another embodiment, the compound for use in the present
invention is represented by the structure of the general formula
(VI):
##STR00018##
[0241] wherein [0242] R.sub.1 is either hydrogen or a linear,
saturated, mono-unsaturated, or poly-unsaturated, alkyl chain
ranging in length from 2 to 30 carbon atoms; [0243] R.sub.2 is a
linear, saturated, mono-unsaturated, or poly-unsaturated, alkyl
chain ranging in length from 2 to 30 carbon atoms; [0244] Z is
either nothing, inositol, choline, or glycerol; [0245] Y is either
nothing or a spacer group ranging in length from 2 to 30 atoms;
[0246] X is a glycosaminoglycan; and [0247] n is a number from 1 to
1000; [0248] wherein any bond between the phospholipid, Z, Y and X
is either an amide or an esteric bond.
[0249] In another embodiment, the compound for use in the present
invention is represented by the structure of the general formula
(VI):
##STR00019##
[0250] wherein [0251] R.sub.1 is either hydrogen or a linear,
saturated, mono-unsaturated, or poly-unsaturated, alkyl chain
ranging in length from 2 to 30 carbon atoms; [0252] R.sub.2 is a
linear, saturated, mono-unsaturated, or poly-unsaturated, alkyl
chain ranging in length from 2 to 30 carbon atoms; [0253] Z is
either nothing, inositol, choline, or glycerol; [0254] Y is either
nothing or a spacer group ranging in length from 2 to 30 atoms;
[0255] X is a glycosaminoglycan; and [0256] n is a number from 2 to
1000; [0257] wherein any bond between the phospholipid, Z, Y and X
is either an amide or an esteric bond.
[0258] In another embodiment, the compound for use in the present
invention is represented by the structure of the general formula
(VII):
##STR00020##
[0259] wherein [0260] R.sub.1 is a linear, saturated,
mono-unsaturated, or poly-unsaturated, alkyl chain ranging in
length from 2 to 30 carbon atoms; [0261] R.sub.2 is either hydrogen
or a linear, saturated, mono-unsaturated, or poly-unsaturated,
alkyl chain ranging in length from 2 to 30 carbon atoms; [0262] Z
is either nothing, inositol, choline, or glycerol; [0263] Y is
either nothing or a spacer group ranging in length from 2 to 30
atoms; [0264] X is a glycosaminoglycan; and [0265] n is a number
from 1 to 1000; [0266] wherein any bond between the phospholipid,
Z, Y and X is either an amide or an esteric bond.
[0267] In another embodiment, the compound for use in the present
invention is represented by the structure of the general formula
(VII):
##STR00021##
[0268] wherein [0269] R.sub.1 is a linear, saturated,
mono-unsaturated, or poly-unsaturated, alkyl chain ranging in
length from 2 to 30 carbon atoms; [0270] R.sub.2 is either hydrogen
or a linear, saturated, mono-unsaturated, or poly-unsaturated,
alkyl chain ranging in length from 2 to 30 carbon atoms; [0271] Z
is either nothing, inositol, choline, or glycerol; [0272] Y is
either nothing or a spacer group ranging in length from 2 to 30
atoms; [0273] X is a glycosaminoglycan; and [0274] n is a number
from 1 to 1000; [0275] wherein any bond between the phospholipid,
Z, Y and X is either an amide or an esteric bond.
[0276] In one embodiment of the invention, phosphatidylcholine
(PC), phosphatidylinositol (PI), phosphatidic acid (PA), wherein Z
is nothing, and phosphatidylglycerol (PG) conjugates are herein
defined as compounds of the general formula (III).
[0277] In another embodiment, the compound for use in the present
invention is represented by the structure of the general formula
(VIII):
##STR00022##
[0278] wherein [0279] R.sub.1 is a linear, saturated,
mono-unsaturated, or poly-unsaturated, alkyl chain ranging in
length from 2 to 30 carbon atoms; [0280] R.sub.2 is either hydrogen
or a linear, saturated, mono-unsaturated, or poly-unsaturated,
alkyl chain ranging in length from 2 to 30 carbon atoms; [0281] Z
is either nothing, ethanolamine, serine, inositol, choline, or
glycerol; [0282] Y is either nothing or a spacer group ranging in
length from 2 to 30 atoms; [0283] X is a glycosaminoglycan; and
[0284] n is a number from 1 to 1000; [0285] wherein any bond
between the phospholipid, Z, Y and X is either an amide or an
esteric bond.
[0286] In another embodiment, the compound for use in the present
invention is represented by the structure of the general formula
(VIII):
##STR00023##
[0287] wherein [0288] R.sub.1 is a linear, saturated,
mono-unsaturated, or poly-unsaturated, alkyl chain ranging in
length from 2 to 30 carbon atoms; [0289] R.sub.2 is either hydrogen
or a linear, saturated, mono-unsaturated, or poly-unsaturated,
alkyl chain ranging in length from 2 to 30 carbon atoms; [0290] Z
is either nothing, ethanolamine, serine, inositol, choline, or
glycerol; [0291] Y is either nothing or a spacer group ranging in
length from 2 to 30 atoms; [0292] X is a glycosaminoglycan; and
[0293] n is a number from 2 to 1000; [0294] wherein any bond
between the phospholipid, Z, Y and X is either an amide or an
esteric bond.
[0295] In another embodiment, the compound for use in the present
invention is represented by the structure of the general formula
(IX):
##STR00024##
[0296] wherein [0297] R.sub.1 is either hydrogen or a linear,
saturated, mono-unsaturated, or poly-unsaturated, alkyl chain
ranging in length from 2 to 30 carbon atoms; [0298] R.sub.2 is
either hydrogen or a linear, saturated, mono-unsaturated, or
poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon
atoms; [0299] Z is either nothing, ethanolamine, serine, inositol,
choline, or glycerol; [0300] Y is either nothing or a spacer group
ranging in length from 2 to 30 atoms; [0301] X is a
glycosaminoglycan; and [0302] n is a number from 1 to 1000; [0303]
wherein any bond between the phospholipid, Z, Y and X is either an
amide or an esteric bond.
[0304] In another embodiment, the compound for use in the present
invention is represented by the structure of the general formula
(IX):
##STR00025##
[0305] wherein [0306] R.sub.1 is either hydrogen or a linear,
saturated, mono-unsaturated, or poly-unsaturated, alkyl chain
ranging in length from 2 to 30 carbon atoms; [0307] R.sub.2 is
either hydrogen or a linear, saturated, mono-unsaturated, or
poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon
atoms; [0308] Z is either nothing, ethanolamine, serine, inositol,
choline, or glycerol; [0309] Y is either nothing or a spacer group
ranging in length from 2 to 30 atoms; [0310] X is a
glycosaminoglycan; and [0311] n is a number from 2 to 1000; [0312]
wherein any bond between the phospholipid, Z, Y and X is either an
amide or an esteric bond.
[0313] In another embodiment, the compound for use in the present
invention is represented by the structure of the general formula
(IXa):
##STR00026##
[0314] wherein [0315] R.sub.1 is either hydrogen or a linear,
saturated, mono-unsaturated, or poly-unsaturated, alkyl chain
ranging in length from 2 to 30 carbon atoms; [0316] R.sub.2 is
either hydrogen or a linear, saturated, mono-unsaturated, or
poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon
atoms; [0317] Z is either nothing, ethanolamine, serine, inositol,
choline, or glycerol; [0318] Y is either nothing or a spacer group
ranging in length from 2 to 30 atoms; [0319] X is a
glycosaminoglycan; and [0320] n is a number from 1 to 1000; [0321]
wherein any bond between the phospholipid, Z, Y and X is either an
amide or an esteric bond.
[0322] In another embodiment, the compound for use in the present
invention is represented by the structure of the general formula
(IXa):
##STR00027##
[0323] wherein [0324] R.sub.1 is either hydrogen or a linear,
saturated, mono-unsaturated, or poly-unsaturated, alkyl chain
ranging in length from 2 to 30 carbon atoms; [0325] R.sub.2 is
either hydrogen or a linear, saturated, mono-unsaturated, or
poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon
atoms; [0326] Z is either nothing, ethanolamine, serine, inositol,
choline, or glycerol; [0327] Y is either nothing or a spacer group
ranging in length from 2 to 30 atoms; [0328] X is a
glycosaminoglycan; and [0329] n is a number from 2 to 1000; [0330]
wherein any bond between the phospholipid, Z, Y and X is either an
amide or an esteric bond.
[0331] In another embodiment, the compound for use in the present
invention is represented by the structure of the general formula
(IXb):
##STR00028##
[0332] wherein [0333] R.sub.1 is either hydrogen or a linear,
saturated, mono-unsaturated, or poly-unsaturated, alkyl chain
ranging in length from 2 to 30 carbon atoms; [0334] R.sub.2 is
either hydrogen or a linear, saturated, mono-unsaturated, or
poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon
atoms; [0335] Z is either nothing, ethanolamine, serine, inositol,
choline, or glycerol; [0336] Y is either nothing or a spacer group
ranging in length from 2 to 30 atoms; [0337] X is a
glycosaminoglycan; and [0338] n is a number from 1 to 1000; [0339]
wherein any bond between the phospholipid, Z, Y and X is either an
amide or an esteric bond.
[0340] In another embodiment, the compound for use in the present
invention is represented by the structure of the general formula
(IXb):
##STR00029##
[0341] wherein [0342] R.sub.1 is either hydrogen or a linear,
saturated, mono-unsaturated, or poly-unsaturated, alkyl chain
ranging in length from 2 to 30 carbon atoms; [0343] R.sub.2 is
either hydrogen or a linear, saturated, mono-unsaturated, or
poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon
atoms; [0344] Z is either nothing, ethanolamine, serine, inositol,
choline, or glycerol; [0345] Y is either nothing or a spacer group
ranging in length from 2 to 30 atoms; [0346] X is a
glycosaminoglycan; and [0347] n is a number from 2 to 1000; [0348]
wherein any bond between the phospholipid, Z, Y and X is either an
amide or an esteric bond.
[0349] In another embodiment, the compound for use in the present
invention is represented by the structure of the general formula
(X):
##STR00030##
[0350] wherein [0351] R.sub.1 is either hydrogen or a linear,
saturated, mono-unsaturated, or poly-unsaturated, alkyl chain
ranging in length from 2 to 30 carbon atoms; [0352] R.sub.2 is a
linear, saturated, mono-unsaturated, or poly-unsaturated, alkyl
chain ranging in length from 2 to 30 carbon atoms; [0353] Z is
either nothing, ethanolamine, serine, inositol, choline, or
glycerol; [0354] Y is either nothing or a spacer group ranging in
length from 2 to 30 atoms; [0355] X is a glycosaminoglycan; and
[0356] n is a number from 1 to 1000; [0357] wherein any bond
between the ceramide phosphoryl, Z, Y and X is either an amide or
an esteric bond.
[0358] In another embodiment, the compound for use in the present
invention is represented by the structure of the general formula
(X):
##STR00031##
[0359] wherein [0360] R.sub.1 is either hydrogen or a linear,
saturated, mono-unsaturated, or poly-unsaturated, alkyl chain
ranging in length from 2 to 30 carbon atoms; [0361] R.sub.2 is a
linear, saturated, mono-unsaturated, or poly-unsaturated, alkyl
chain ranging in length from 2 to 30 carbon atoms; [0362] Z is
either nothing, ethanolamine, serine, inositol, choline, or
glycerol; [0363] Y is either nothing or a spacer group ranging in
length from 2 to 30 atoms; [0364] X is a glycosaminoglycan; and
[0365] n is a number from 2 to 1000;
[0366] wherein any bond between the ceramide phosphoryl, Z, Y and X
is either an amide or an esteric bond.
[0367] In another embodiment, the compound for use in the present
invention is represented by the structure of the general formula
(XI):
##STR00032##
[0368] wherein [0369] R.sub.1 is a linear, saturated,
mono-unsaturated, or poly-unsaturated, alkyl chain ranging in
length from 2 to 30 carbon atoms; [0370] Y is either nothing or a
spacer group ranging in length from 2 to 30 atoms; [0371] X is a
glycosaminoglycan; and [0372] n is a number from 1 to 1000; [0373]
wherein if Y is nothing the sphingosyl is directly linked to X via
an amide bond and if Y is a spacer, the spacer is directly linked
to X and to the sphingosyl via an amide bond and to X via an amide
or an esteric bond.
[0374] In another embodiment, the compound for use in the present
invention is represented by the structure of the general formula
(XI):
##STR00033##
[0375] wherein [0376] R.sub.1 is a linear, saturated,
mono-unsaturated, or poly-unsaturated, alkyl chain ranging in
length from 2 to 30 carbon atoms; [0377] Y is either nothing or a
spacer group ranging in length from 2 to 30 atoms; [0378] X is a
glycosaminoglycan; and [0379] n is a number from 2 to 1000; [0380]
wherein if Y is nothing the sphingosyl is directly linked to X via
an amide bond and if Y is a spacer, the spacer is directly linked
to X and to the sphingosyl via an amide bond and to X via an amide
or an esteric bond.
[0381] In another embodiment, the compound for use in the present
invention is represented by the structure of the general formula
(XII):
##STR00034##
[0382] wherein [0383] R.sub.1 is a linear, saturated,
mono-unsaturated, or poly-unsaturated, alkyl chain ranging in
length from 2 to 30 carbon atoms; [0384] R.sub.2 is a linear,
saturated, mono-unsaturated, or poly-unsaturated, alkyl chain
ranging in length from 2 to 30 carbon atoms; [0385] Z is either
nothing, ethanolamine, serine, inositol, choline, or glycerol;
[0386] Y is either nothing or a spacer group ranging in length from
2 to 30 atoms; [0387] X is a glycosaminoglycan; and [0388] n is a
number from 1 to 1000; [0389] wherein any bond between the
ceramide, Z, Y and X is either an amide or an esteric bond.
[0390] In another embodiment, the compound for use in the present
invention is represented by the structure of the general formula
(XII):
##STR00035##
[0391] wherein [0392] R.sub.1 is a linear, saturated,
mono-unsaturated, or poly-unsaturated, alkyl chain ranging in
length from 2 to 30 carbon atoms; [0393] R.sub.2 is a linear,
saturated, mono-unsaturated, or poly-unsaturated, alkyl chain
ranging in length from 2 to 30 carbon atoms; [0394] Z is either
nothing, ethanolamine, serine, inositol, choline, or glycerol;
[0395] Y is either nothing or a spacer group ranging in length from
2 to 30 atoms; [0396] X is a glycosaminoglycan; and [0397] n is a
number from 2 to 1000; [0398] wherein any bond between the
ceramide, Z, Y and X is either an amide or an esteric bond.
[0399] In another embodiment, the compound for use in the present
invention is represented by the structure of the general formula
(XIII):
##STR00036##
(XIII)
[0400] wherein [0401] R.sub.1 is a linear, saturated,
mono-unsaturated, or poly-unsaturated, alkyl chain ranging in
length from 2 to 30 carbon atoms; [0402] R.sub.2 is a linear,
saturated, mono-unsaturated, or poly-unsaturated, alkyl chain
ranging in length from 2 to 30 carbon atoms; [0403] Z is either
nothing, choline, phosphate, inositol, or glycerol; [0404] Y is
either nothing or a spacer group ranging in length from 2 to 30
atoms; [0405] X is a glycosaminoglycan; and [0406] n is a number
from 1 to 1000; [0407] wherein any bond between the diglyceryl, Z,
Y and X is either an amide or an esteric bond.
[0408] In another embodiment, the compound for use in the present
invention is represented by the structure of the general formula
(XIII):
##STR00037##
[0409] wherein [0410] R.sub.1 is a linear, saturated,
mono-unsaturated, or poly-unsaturated, alkyl chain ranging in
length from 2 to 30 carbon atoms; [0411] R.sub.2 is a linear,
saturated, mono-unsaturated, or poly-unsaturated, alkyl chain
ranging in length from 2 to 30 carbon atoms; [0412] Z is either
nothing, choline, phosphate, inositol, or glycerol; [0413] Y is
either nothing or a spacer group ranging in length from 2 to 30
atoms; [0414] X is a glycosaminoglycan; and [0415] n is a number
from 2 to 1000; [0416] wherein any bond between the diglyceryl, Z,
Y and X is either an amide or an esteric bond.
[0417] In another embodiment, the compound for use in the present
invention is represented by the structure of the general formula
(XIV):
##STR00038##
[0418] wherein [0419] R.sub.1 is either hydrogen or a linear,
saturated, mono-unsaturated, or poly-unsaturated, alkyl chain
ranging in length from 2 to 30 carbon atoms; [0420] R.sub.2 is a
linear, saturated, mono-unsaturated, or poly-unsaturated, alkyl
chain ranging in length from 2 to 30 carbon atoms; [0421] Z is
either nothing, choline, phosphate, inositol, or glycerol; [0422] Y
is either nothing or a spacer group ranging in length from 2 to 30
atoms; [0423] X is a glycosaminoglycan; and [0424] n is a number
from 1 to 1000; [0425] wherein any bond between the glycerolipid,
Z, Y and X is either an amide or an esteric bond.
[0426] In another embodiment, the compound for use in the present
invention is represented by the structure of the general formula
(XIV):
##STR00039##
[0427] wherein [0428] R.sub.1 is either hydrogen or a linear,
saturated, mono-unsaturated, or poly-unsaturated, alkyl chain
ranging in length from 2 to 30 carbon atoms; [0429] R.sub.2 is a
linear, saturated, mono-unsaturated, or poly-unsaturated, alkyl
chain ranging in length from 2 to 30 carbon atoms; [0430] Z is
either nothing, choline, phosphate, inositol, or glycerol; [0431] Y
is either nothing or a spacer group ranging in length from 2 to 30
atoms; [0432] X is a glycosaminoglycan; and [0433] n is a number
from 2 to 1000; [0434] wherein any bond between the glycerolipid,
Z, Y and X is either an amide or an esteric bond.
[0435] In another embodiment, the compound for use in the present
invention is represented by the structure of the general formula
(XV):
##STR00040##
[0436] wherein [0437] R.sub.1 is a linear, saturated,
mono-unsaturated, or poly-unsaturated, alkyl chain ranging in
length from 2 to 30 carbon atoms; [0438] R.sub.2 is either hydrogen
or a linear, saturated, mono-unsaturated, or poly-unsaturated,
alkyl chain ranging in length from 2 to 30 carbon atoms; [0439] Z
is either nothing, choline, phosphate, inositol, or glycerol;
[0440] Y is either nothing or a spacer group ranging in length from
2 to 30 atoms; [0441] X is a glycosaminoglycan; and [0442] n is a
number from 1 to 1000; [0443] wherein any bond between the
glycerolipid, Z, Y and X is either an amide or an esteric bond.
[0444] In another embodiment, the compound for use in the present
invention is represented by the structure of the general formula
(XV):
##STR00041##
[0445] wherein [0446] R.sub.1 is a linear, saturated,
mono-unsaturated, or poly-unsaturated, alkyl chain ranging in
length from 2 to 30 carbon atoms; [0447] R.sub.2 is either hydrogen
or a linear, saturated, mono-unsaturated, or poly-unsaturated,
alkyl chain ranging in length from 2 to 30 carbon atoms; [0448] Z
is either nothing, choline, phosphate, inositol, or glycerol;
[0449] Y is either nothing or a spacer group ranging in length from
2 to 30 atoms; [0450] X is a glycosaminoglycan; and [0451] n is a
number from 2 to 1000; [0452] wherein any bond between the
glycerolipid, Z, Y and X is either an amide or an esteric bond.
[0453] In another embodiment, the compound for use in the present
invention is represented by the structure of the general formula
(XVI):
##STR00042##
[0454] wherein [0455] R.sub.1 is either hydrogen or a linear,
saturated, mono-unsaturated, or poly-unsaturated, alkyl chain
ranging in length from 2 to 30 carbon atoms; [0456] R.sub.2 is a
linear, saturated, mono-unsaturated, or poly-unsaturated, alkyl
chain ranging in length from 2 to 30 carbon atoms; [0457] Z is
either nothing, choline, phosphate, inositol, or glycerol; [0458] Y
is either nothing or a spacer group ranging in length from 2 to 30
atoms; [0459] X is a glycosaminoglycan; and [0460] n is a number
from 1 to 1000; [0461] wherein any bond between the lipid, Z, Y and
X is either an amide or an esteric bond.
[0462] In another embodiment, the compound for use in the present
invention is represented by the structure of the general formula
(XVI):
##STR00043##
[0463] wherein [0464] R.sub.1 is either hydrogen or a linear,
saturated, mono-unsaturated, or poly-unsaturated, alkyl chain
ranging in length from 2 to 30 carbon atoms; [0465] R.sub.2 is a
linear, saturated, mono-unsaturated, or poly-unsaturated, alkyl
chain ranging in length from 2 to 30 carbon atoms; [0466] Z is
either nothing, choline, phosphate, inositol, or glycerol; [0467] Y
is either nothing or a spacer group ranging in length from 2 to 30
atoms; [0468] X is a glycosaminoglycan; and [0469] n is a number
from 2 to 1000; [0470] wherein any bond between the lipid, Z, Y and
X is either an amide or an esteric bond.
[0471] In another embodiment, the compound for use in the present
invention is represented by the structure of the general formula
(XVII):
##STR00044##
[0472] wherein [0473] R.sub.1 is either hydrogen or a linear,
saturated, mono-unsaturated, or poly-unsaturated, alkyl chain
ranging in length from 2 to 30 carbon atoms; [0474] R.sub.2 is a
linear, saturated, mono-unsaturated, or poly-unsaturated, alkyl
chain ranging in length from 2 to 30 carbon atoms; [0475] Z is
either nothing, choline, phosphate, inositol, or glycerol; [0476] Y
is either nothing or a spacer group ranging in length from 2 to 30
atoms; [0477] X is a glycosaminoglycan; and [0478] n is a number
from 1 to 1000; [0479] wherein any bond between the lipid, Z, Y and
X is either an amide or an esteric bond.
[0480] In another embodiment, the compound for use in the present
invention is represented by the structure of the general formula
(XVII):
##STR00045##
[0481] wherein [0482] R.sub.1 is either hydrogen or a linear,
saturated, mono-unsaturated, or poly-unsaturated, alkyl chain
ranging in length from 2 to 30 carbon atoms; [0483] R.sub.2 is a
linear, saturated, mono-unsaturated, or poly-unsaturated, alkyl
chain ranging in length from 2 to 30 carbon atoms; [0484] Z is
either nothing, choline, phosphate, inositol, or glycerol; [0485] Y
is either nothing or a spacer group ranging in length from 2 to 30
atoms; [0486] X is a glycosaminoglycan; and [0487] n is a number
from 2 to 1000; [0488] wherein any bond between the lipid, Z, Y and
X is either an amide or an esteric bond.
[0489] In another embodiment, the compound for use in the present
invention is represented by the structure of the general formula
(XVIII):
##STR00046##
[0490] wherein [0491] R.sub.1 is either hydrogen or a linear,
saturated, mono-unsaturated, or poly-unsaturated, alkyl chain
ranging in length from 2 to 30 carbon atoms; [0492] R.sub.2 is
either hydrogen or a linear, saturated, mono-unsaturated, or
poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon
atoms; [0493] Z is either nothing, choline, phosphate, inositol, or
glycerol; [0494] Y is either nothing or a spacer group ranging in
length from 2 to 30 atoms; [0495] X is a glycosaminoglycan; and
[0496] n is a number from 1 to 1000; [0497] wherein any bond
between the lipid, Z, Y and X is either an amide or an esteric
bond.
[0498] In another embodiment, the compound for use in the present
invention is represented by the structure of the general formula
(XVIII):
##STR00047##
[0499] wherein [0500] R.sub.1 is either hydrogen or a linear,
saturated, mono-unsaturated, or poly-unsaturated, alkyl chain
ranging in length from 2 to 30 carbon atoms; [0501] R.sub.2 is
either hydrogen or a linear, saturated, mono-unsaturated, or
poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon
atoms; [0502] Z is either nothing, choline, phosphate, inositol, or
glycerol; [0503] Y is either nothing or a spacer group ranging in
length from 2 to 30 atoms; [0504] X is a glycosaminoglycan; and
[0505] n is a number from 2 to 1000; [0506] wherein any bond
between the lipid, Z, Y and X is either an amide or an esteric
bond.
[0507] In another embodiment, the compound for use in the present
invention is represented by the structure of the general formula
(XIX):
##STR00048##
[0508] wherein [0509] R.sub.1 is either hydrogen or a linear,
saturated, mono-unsaturated, or poly-unsaturated, alkyl chain
ranging in length from 2 to 30 carbon atoms; [0510] R.sub.2 is
either hydrogen or a linear, saturated, mono-unsaturated, or
poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon
atoms; [0511] Z is either nothing, choline, phosphate, inositol, or
glycerol; [0512] Y is either nothing or a spacer group ranging in
length from 2 to 30 atoms; [0513] X is a glycosaminoglycan; and
[0514] n is a number from 1 to 1000; [0515] wherein any bond
between the lipid, Z, Y and X is either an amide or an esteric
bond.
[0516] In another embodiment, the compound for use in the present
invention is represented by the structure of the general formula
(XIX):
##STR00049##
[0517] wherein [0518] R.sub.1 is either hydrogen or a linear,
saturated, mono-unsaturated, or poly-unsaturated, alkyl chain
ranging in length from 2 to 30 carbon atoms; [0519] R.sub.2 is
either hydrogen or a linear, saturated, mono-unsaturated, or
poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon
atoms; [0520] Z is either nothing, choline, phosphate, inositol, or
glycerol; [0521] Y is either nothing or a spacer group ranging in
length from 2 to 30 atoms; [0522] X is a glycosaminoglycan; and
[0523] n is a number from 2 to 1000; [0524] wherein any bond
between the lipid, Z, Y and X is either an amide or an esteric
bond.
[0525] In another embodiment, the compound for use in the present
invention is represented by the structure of the general formula
(XX):
##STR00050##
[0526] wherein [0527] R.sub.1 is either hydrogen or a linear,
saturated, mono-unsaturated, or poly-unsaturated, alkyl chain
ranging in length from 2 to 30 carbon atoms; [0528] R.sub.2 is
either hydrogen or a linear, saturated, mono-unsaturated, or
poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon
atoms; [0529] Z is either nothing, choline, phosphate, inositol, or
glycerol; [0530] Y is either nothing or a spacer group ranging in
length from 2 to 30 atoms; [0531] X is a glycosaminoglycan; and
[0532] n is a number from 1 to 1000; [0533] wherein any bond
between the lipid, Z, Y and X is either an amide or an esteric
bond.
[0534] In another embodiment, the compound for use in the present
invention is represented by the structure of the general formula
(XX):
##STR00051##
[0535] wherein [0536] R.sub.1 is either hydrogen or a linear,
saturated, mono-unsaturated, or poly-unsaturated, alkyl chain
ranging in length from 2 to 30 carbon atoms; [0537] R.sub.2 is
either hydrogen or a linear, saturated, mono-unsaturated, or
poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon
atoms; [0538] Z is either nothing, choline, phosphate, inositol, or
glycerol; [0539] Y is either nothing or a spacer group ranging in
length from 2 to 30 atoms; [0540] X is a glycosaminoglycan; and
[0541] n is a number from 2 to 1000; [0542] wherein any bond
between the lipid, Z, Y and X is either an amide or an esteric
bond.
[0543] In another embodiment, the compound for use in the present
invention is represented by the structure of the general formula
(XXI):
##STR00052##
[0544] wherein [0545] R.sub.1 is either hydrogen or a linear,
saturated, mono-unsaturated, or poly-unsaturated, alkyl chain
ranging in length from 2 to 30 carbon atoms; [0546] R.sub.2 is
either hydrogen or a linear, saturated, mono-unsaturated, or
poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon
atoms; [0547] Z is either nothing, choline, phosphate, inositol, or
glycerol; [0548] Y is either nothing or a spacer group ranging in
length from 2 to 30 atoms; [0549] X is a glycosaminoglycan; and
[0550] n is a number from 1 to 1000; [0551] wherein any bond
between the lipid, Z, Y and X is either an amide or an esteric
bond.
[0552] In another embodiment, the compound for use in the present
invention is represented by the structure of the general formula
(XXI):
##STR00053##
[0553] wherein [0554] R.sub.1 is either hydrogen or a linear,
saturated, mono-unsaturated, or poly-unsaturated, alkyl chain
ranging in length from 2 to 30 carbon atoms; [0555] R.sub.2 is
either hydrogen or a linear, saturated, mono-unsaturated, or
poly-unsaturated, alkyl chain ranging in length from 2 to 30 carbon
atoms; [0556] Z is either nothing, choline, phosphate, inositol, or
glycerol; [0557] Y is either nothing or a spacer group ranging in
length from 2 to 30 atoms; [0558] X is a glycosaminoglycan; and
[0559] n is a number from 2 to 1000; [0560] wherein any bond
between the lipid, Z, Y and X is either an amide or an esteric
bond.
[0561] For any or all of the compounds represented by the
structures of the general formulae (A), (I), (II), (III), (IV),
(V), (VI), (VII), (VIII), (IX), (IXa), (IXb), (X), (XI), (XII),
(XIII), (XIV), (XV), (XVI), (XVII), (XVIII), (XIX), (XX), (XXI),
and (XXII) hereinabove: In one embodiment, X is a
glycosaminoglycan. According to this aspect and in one embodiment,
the glycosaminoglycan may be, inter alia, hyaluronic acid, heparin,
heparan sulfate, chondroitin sulfate, keratin, keratan sulfate,
dermatan sulfate or a derivative thereof. In another embodiment, X
is not a glycosaminoglycan. In another embodiment, X is a
polysaccharide, which in one embodiment is a hetero-polysaccharide,
and in another embodiment, is a homo-polysaccharide. In another
embodiment, X is a polypyranose.
[0562] In another embodiment, the glycosaminoglycan is a polymer of
disaccharide units. In another embodiment, the number of the
disaccharide units in the polymer is m. In another embodiment, m is
a number from 2-10,000. In another embodiment, m is a number from
2-500. In another embodiment, m is a number from 2-1000. In another
embodiment, m is a number from 50-500. In another embodiment, m is
a number from 2-2000. In another embodiment, m is a number from
500-2000. In another embodiment, m is a number from 1000-2000. In
another embodiment, m is a number from 2000-5000. In another
embodiment, m is a number from 3000-7000. In another embodiment, m
is a number from 5000-10,000. In another embodiment, a disaccharide
unit of a glycosaminoglycan may be bound to one lipid or
phospholipid moiety. In another embodiment, each disaccharide unit
of the glycosaminoglycan may be bound to zero or one lipid or
phospholipid moieties. In another embodiment, the lipid or
phospholipid moieties are bound to the --COOH group of the
disaccharide unit. In another embodiment, the bond between the
lipid or phospholipid moiety and the disaccharide unit is an amide
bond.
[0563] In one embodiment, the invention encompasses compounds,
compositions and preparations, comprising a phospholipid-GAG
conjugate, whereby the molar ratio in the compounds, compositions
and preparations between the phospholipid and the GAG is in the
range of between 1.5:1 to 20:1, or in another embodiments, 2:1 to
10:1, or in another embodiment, 3:1 to 7:1, or in another
embodiment, 1.5:1 to 7:1. In another embodiment, the molar ratio
between the phospholipid and the GAG is in the range of between 2:1
to 10:1. In another embodiment, the molar ratio between the
phospholipid and the GAG is in the range of between 2:1 to 5:1. In
another embodiment, the molar ration between the phospholipid and
the GAG is 2:1. In another embodiment, the molar ration between the
phospholipid and the GAG is 3:1. In another embodiment, the molar
ration between the phospholipid and the GAG is 5:1. In another
embodiment, the molar ration between the phospholipid and the GAG
is 10:1. In another embodiment, the molar ration between the
phospholipid and the GAG is 20:1.
[0564] In one embodiment, the compound of the present invention
comprises a glycosaminoglycan (GAG) with a molecular weight in the
range of between 30-100 kD. In another embodiment, the GAG has a
molecular weight in the range of between 30-80 kD. In another
embodiment, the GAG has a molecular weight in the range of between
30-50 kD. In another embodiment, the GAG has a molecular weight in
the range of between 20-80 kD. In another embodiment, the GAG has a
molecular weight in the range of between 20-50 kD.
[0565] In another embodiment, the chondroitin sulfate may be, inter
alia, chondroitin-6-sulfate, chondroitin-4-sulfate or a derivative
thereof.
[0566] In one embodiment of the invention, Y is nothing.
Non-limiting examples of suitable divalent groups forming the
optional bridging group (which in one embodiment, is referred to as
a spacer) Y, according to embodiments of the invention, are
straight or branched chain alkylene, e.g. of 2 or more, preferably
4 to 30 carbon atoms, --CO-alkylene-CO, --NH-alkylene-NH--,
--CO-alkylene-NH--, --NH-alkylene-NH, CO-alkylene-NH--, an amino
acid, cycloalkylene, wherein alkylene in each instance, is straight
or branched chain and contains 2 or more, preferably 2 to 30 atoms
in the chain, --(--O--CH(CH.sub.3)CH.sub.2).sub.x wherein x is an
integer of 1 or more.
[0567] According to embodiments of the invention, in addition to
the traditional phospholipid structure, related derivatives for use
in this invention are phospholipids modified at the C1 or C2
position to contain an amine, ether or alkyl bond instead of an
ester bond. In one embodiment of the invention, the alkyl
phospholipid derivatives and ether phospholipid derivatives are
exemplified herein.
[0568] In one embodiment of the invention, the sugar rings of the
glycosaminoglycan are intact. In another embodiment, intact refers
to closed. In another embodiment, intact refers to natural. In
another embodiment, intact refers to unbroken.
[0569] In one embodiment of the invention, the structure of the
lipid or phospholipid in any compound according to the invention is
intact. In another embodiment, the natural structure of the lipid
or phospholipids in any compound according to the invention is
maintained.
[0570] In one embodiment, the compounds for use in the present
invention are biodegradable.
[0571] In one embodiment, the compound according to the invention
is phosphatidylethanolamine bound to aspirin. In one embodiment,
the compound according to the invention is phosphatidylethanolamine
bound to glutarate.
[0572] In some embodiments, the compounds for use are as listed in
Table 1 below.
TABLE-US-00001 TABLE 1 Phospholipid Spacer Polymer (m.w.) Compound
PE None Hyaluronic acid (2-2000 kDa) XXII Dimyristoyl-PE None
Hyaluronic acid XXIII PE None Heparin (0.5-110 kDa) XXIV PE None
Chondroitin sulfate A XXV PE None Carboxymethylcellulose (20-500
kDa) XXVI PE Dicarboxylic acid + Diamine Polygeline (haemaccel)
(4-40 kDa) XXVII PE None Hydroxyethylstarch XXVIII PE Dicarboxylic
acid + Diamine Dextran (1-2,000 kDa) XXIX PE None Aspirin XXX PE
Carboxyl amino group Hyaluronic acid (2-2000 kDa) XXXI PE
Dicarboxyl group Hyaluronic acid (2-2000 kDa) XXXII PE Dipalmitoic
acid Hyaluronic acid (2-2000 kDa) XXXIII PE Carboxyl amino group
Heparin (0.5-110 kDa) XXXIV PE Dicarboxyl group Heparin (0.5-110
kDa) XXXV PE Carboxyl amino group Chondroitin sulfate A XXXVI PE
Dicarboxyl group Chondroitin sulfate A XXXVII PE Carboxyl amino
group Carboxymethylcellulose (20-500 kDa) XXXVIII PE Dicarboxyl
group Carboxymethylcellulose (20-500 kDa) XXXIX PE None Polygeline
(haemaccel) (4-40 kDa) XL PE Carboxyl amino group Polygeline
(haemaccel) (4-40 kDa) XLI PE Dicarboxyl group Polygeline
(haemaccel) (4-40 kDa) XLII PE Carboxyl amino group
Hydroxyethylstarch XLIII PE Dicarboxyl group Hydroxyethylstarch
XLIV PE None Dextran (1-2,000 kDa) XLV PE Carboxyl amino group
Dextran (1-2,000 kDa) XLVI PE Dicarboxyl group Dextran (1-2,000
kDa) XLVII PE Carboxyl amino group Aspirin XLVIII PE Dicarboxyl
group Aspirin XLIX PE None Albumin L PE None Alginate (2-2000 kDa)
LI PE None Polyaminoacid LII PE None Polyethylene glycol LIII PE
None Lactobionic acid LIV PE None Acetylsalicylate LV PE None
Cholesteryl-hemmisuccinate LVI PE None Maltose LVII PE None Cholic
acid LVIII PE None Chondroitin sulfates LIX PE None
Polycarboxylated polyethylene glycol LX Dipalmitoyl-PE None
Hyaluronic acid LXI Dipalmitoyl-PE None Heparin LXII Dipalmitoyl-PE
None Chondroitin sulfate A LXIII Dipalmitoyl-PE None
Carboxymethylcellulose LXIV Dipalmitoyl-PE None Polygeline
(haemaccel) LXV Dipalmitoyl-PE None Hydroxyethylstarch LXVI
Dipalmitoyl-PE None Dextran LXVII Dipalmitoyl-PE None Aspirin
LXVIII Dimyristoyl-PE None Heparin LXVIX Dimyristoyl-PE None
Chondroitin sulfate A LXX Dimyristoyl-PE None
Carboxymethylcellulose LXXI Dimyristoyl-PE None Polygeline
(haemaccel) LXXII Dimyristoyl-PE None Hydroxyethylstarch LXXIII
Dimyristoyl-PE None Dextran LXXIV Dimyristoyl-PE None Aspirin LXXV
PS None Hyaluronic acid LXXVI PS None Heparin LXXVII PS None
Polygeline (haemaccel) LXXVIII PC None Hyaluronic acid LXXIX PC
None Heparin LXXX PC None Polygeline (haemaccel) LXXXI PI None
Hyaluronic acid LXXXII PI None Heparin LXXXIII PI None Polygeline
(haemaccel) LXXXIV PG None Hyaluronic acid LXXXV PG None Heparin
LXXXVI PG None Polygeline (haemaccel) LXXXVII PE None Glutaryl
LXXXVIII Dipalmitoyl-PE None Alginate LXXXIX Dimyristoyl-PE None
Alginate XC PS None Alginate XCI PC None Alginate XCII PI None
Alginate XCIII PG None Alginate XCIV PS None Hydroxyethylstarch XCV
PC None Hydroxyethylstarch XCVI PI None Hydroxyethylstarch XCVII PG
None Hydroxyethylstarch XCVIII PE
--CO--(CH.sub.2).sub.3--CO--NH--(CH.sub.2).sub.6--
Hydroxyethylstarch XCIX PE --CO--CH.sub.2-- Carboxymethylcellulose
C
[0573] In one embodiment of the invention, the compounds for use in
the present invention are any one or more of Compounds I-C. In
another embodiment, the invention provides a composition comprising
any combination of any of the compounds of the invention or the use
of any combination of any of the compounds of the invention. In
another embodiment, the invention provides a composition comprising
Compounds XCIX, C, or a combination thereof and uses thereof. In
another embodiment, the invention provides a composition comprising
Compounds LXV, LXVI, LXXI, LXXII, LXXIII, LXXXIX, XC, or a
combination thereof and uses thereof. In another embodiment, the
compounds for use in the present invention are Compound XXII,
Compound XXIII, Compound XXIV, Compound XXV, Compound XXVI,
Compound XXVII, Compound XXVIII, Compound XXIX, Compound XXX,
Compound LI, or pharmaceutically acceptable salts thereof, in
combination with a physiologically acceptable carrier or solvent.
According to embodiments of the invention, these polymers, when
chosen as the conjugated moiety, may vary in molecular weights from
200 to 2,000,000 Daltons. In one embodiment of the invention, the
molecular weight of the polymer as referred to herein is from 200
to 1000 Daltons. In another embodiment, the molecular weight of the
polymer as referred to herein is from 200 to 1000 Daltons. In
another embodiment, the molecular weight of the polymer as referred
to herein is from 1000 to 5000 Daltons. In another embodiment, the
molecular weight of the polymer as referred to herein is from 5000
to 10,000 Daltons. In another embodiment, the molecular weight of
the polymer as referred to herein is from 10,000 to 20,000 Daltons.
In another embodiment, the molecular weight of the polymer as
referred to herein is from 10,000 to 50,000 Daltons. In another
embodiment, the molecular weight of the polymer as referred to
herein is from 20,000 to 70,000 Daltons. In another embodiment, the
molecular weight of the polymer as referred to herein is from
50,000 to 100,000 Daltons. In another embodiment, the molecular
weight of the polymer as referred to herein is from 100,000 to
200,000 Daltons. In another embodiment, the molecular weight of the
polymer as referred to herein is from 200,000 to 500,000 Daltons.
In another embodiment, the molecular weight of the polymer as
referred to herein is from 200,000 to 1,000,000 Daltons. In another
embodiment, the molecular weight of the polymer as referred to
herein is from 500,000 to 1,000,000 Daltons. In another embodiment,
the molecular weight of the polymer as referred to herein is from
1,000,000 to 2,000,000 Daltons. Various molecular weight species
have been shown to have the desired biological efficacy.
[0574] In one embodiment, AlgPE has a molecular weight of
approximately 120 kD, CSAPE has a molecular weight of approximately
100 kD, HemPE has a molecular weight of approximately 75 kD,
HesDMPE has a molecular weight of approximately 90 kD, CMPE has a
molecular weight of approximately 75 kD, or a combination thereof.
In one embodiment, "approximately" refers to up to 5%, 10%, 15%,
20%, or 25% of the value. In another embodiment, "approximately"
refers to 5-25%, 5-15%. 10-25%, 10-20%, 15-25% of the value.
[0575] In one embodiment of this invention, low molecular weight
compounds for use in the present invention are defined hereinabove
as the compounds of formula (I)-(XXI) wherein X is a mono- or
disaccharide, carboxylated disaccharide, mono- or dicarboxylic
acids, a salicylate, salicylic acid, aspirin, lactobionic acid,
maltose, an amino acid, glycine, acetic acid, butyric acid,
dicarboxylic acid, glutaric acid, succinic acid, fatty acid,
dodecanoic acid, didodecanoic acid, bile acid, cholic acid,
cholesterylhemmisuccinate, a di- or tripeptide, an oligopeptide, a
trisacharide, or a di- or trisaccharide monomer unit of heparin,
heparan sulfate, keratin, keratan sulfate, chondroitin,
chondroitin-6-sulfate, chondroitin-4-sulfate, dermatin, dermatan
sulfate, dextran, hyaluronic acid, glycosaminoglycan, or
polypyranose.
[0576] Examples of suitable divalent groups forming the optional
bridging group Y are straight- or branched-chain alkylene, e.g. of
2 or more, preferably 4 to 18 carbon atoms, --CO-alkylene-CO,
--NH-alkylene-NH--, --CO-alkylene-NH--, cycloalkylene, wherein
alkylene in each instance, is straight or branched chain and
contains 2 or more, preferably 2 to 18 carbon atoms in the chain,
--(--O--CH(CH.sub.3)CH.sub.2--).sub.x-- wherein x is an integer of
1 or more.
[0577] In another embodiment, in addition to the traditional
phospholipid structure, related derivatives for use in this
invention are phospholipids modified at the C1 or C2 position to
contain an ether or alkyl bond instead of an ester bond. These
derivatives are exemplified hereinabove by the general formulae
(VIII) and (IX).
[0578] In one embodiment of the invention, X is covalently
conjugated to a lipid. In another embodiment, X is covalently
conjugated to a lipid via an amide bond. In another embodiment, X
is covalently conjugated to a lipid via an esteric bond. In another
embodiment, the lipid is phosphatidylethanolamine.
[0579] In one embodiment, cell surface GAGs play a key role in
protecting cells from diverse damaging agents and processes, such
as reactive oxygen species and free radicals, endotoxins,
cytokines, invasion promoting enzymes, and agents that induce
and/or facilitate degradation of extracellular matrix and basal
membrane, cell invasiveness, white cell extravasation and
infiltration, chemotaxis, and others. In addition, cell surface
GAGs protect cells from bacterial, viral and parasitic infection,
and their stripping exposes the cell to interaction and subsequent
internalization of the microorganism. Enrichment of cell surface
GAGs would thus assist in protection of the cell from injurious
processes. Thus, in one embodiment of the invention, PLA2
inhibitors are conjugated to GAGs or GAG-mimicking molecules. In
another embodiment, these compounds for use in the present
invention provide wide-range protection from diverse injurious
processes, and are effective in amelioration of diseases that
requires cell protection from injurious biochemical mediators.
[0580] In another embodiment, a GAG-mimicking molecule may be,
inter alia, a negatively charged molecule. In another embodiment, a
GAG-mimicking molecule may be, inter alia, a salicylate derivative.
In another embodiment, a GAG-mimicking molecule may be, inter alia,
a dicarboxylic acid.
[0581] In another embodiment, the invention provides a
pharmaceutical composition for treating a subject suffering from an
eye disease, including a lipid or phospholipid moiety bonded to a
physiologically acceptable monomer, dimer, oligomer, or polymer;
and a pharmaceutically acceptable carrier or excipient.
[0582] In another embodiment, the invention provides a
pharmaceutical composition for treating a subject suffering from an
eye disease, including any one of the compounds for use in the
present invention or any combination thereof; and a
pharmaceutically acceptable carrier or excipient. In another
embodiment, the compounds for use in the present invention include,
inter alia, the compounds represented by the structures of the
general formulae as described hereinbelow: (A), (I), (II), (III),
(IV), (V), (VI), (VII), (VIII), (IX), (IXa), (IXb), (X), (XI),
(XII), (XIII), (XIV), (XV), (XVI), (XVII), (XVIII), (XIX), (XX),
(XXI), (XXII), or any combination thereof.
[0583] The combination of lipids, such as, but not limited to
phosphatidylethanolamine and phosphatidylserine, with additional
monomer or polymer moieties, is thus a practical route to the
production of new drugs for medical purposes, provided that the
resultant chemical composition displays the desired range of
pharmacological properties. In one embodiment, the compounds for
use in the present invention possess a combination of multiple and
potent pharmacological effects in addition to the ability to
inhibit the extracellular form of the enzyme phospholipase A2.
While the pharmacological activity of the compounds for use in the
present invention described herein may be due in part to the nature
of the lipid moiety, the multiple and diverse combination of
pharmacological properties observed for the compounds for use in
the present invention emerges from the ability of the compound
structure to act essentially as several different drugs in one
chemical entity.
[0584] In the cases described herein, the diversity of biological
activities and the effectiveness in disease exhibited by the
compounds for use in the present invention far exceed the
properties anticipated by use of the starting materials themselves,
when administered alone or in combination. However, the
phospholipid conjugate compounds, alone or in combination, are
valuable when used in the methods of treating diseases and
conditions specifically described herein.
[0585] Eye Devices
[0586] It is to be understood that the compounds for use in the
present invention may also be used in combination with any device
which is applied to an eye surface or applied to the internal
regions of the eye. In one embodiment, such a device is a contact
lens, while in other embodiments, it is a corneal prosthetic
device, prosthetic iris implant, scleral lens prosthetic device, an
intra-ocular implant, a scleral buckle, ophthalmic tantalum clip,
ophthalmic conformer, artificial eye, absorbable implant, eye
sphere implant, extraocular orbital implant, keratoprosthesis,
intraocular lens, scleral shell, eye valve implant, or a
combination thereof.
[0587] In one embodiment, the present invention provides a
substrate having a coating on at least a portion of a surface of
said substrate, said coating comprising a lipid or phospholipid
moiety bound to a physiologically acceptable monomer, dimer,
oligomer, or polymer as any of the embodiments describe
hereinabove. In one embodiment, the physiological acceptable
monomer, dimer, oligomer, or polymer is a polypyranose. In one
embodiment, the substrate is a contact lens. In one embodiment, the
substrate is an implant. In one embodiment, the substrate is part
of a device for ophthalmic or ophthamologic use.
[0588] In another embodiment, this invention provides a contact
lens solution comprising the compounds of the instant invention.
Contact lens solutions of the instant invention may comprise, inter
alia, rewetting drops, cleaning solutions, washing solutions,
storage solutions, packing solutions, saline solution, daily
cleaner, multipurpose solution, hydrogen peroxide solution, or a
combination thereof. Any solution which may be used for storage,
preservation, or cleaning of a contact lens or lenses is considered
to be an embodiment of this invention. The safety and tolerability
of solutions comprising the subject compounds as comfort
ingredients, for example, in contact lens packaging solutions, is
exemplified in Example 6. In one embodiment, appliances for use
with the lenses and/or solutions of this invention may be coated
with the compounds for use in the instant invention as described
herein, as well.
[0589] In one embodiment, contact lens solutions of the instant
invention will additionally comprise surfactants, tonicity agents,
viscosity builders, anti-microbials, buffering agents, or a
combination thereof. In one embodiment, surfactants may be
non-ionic, and in one embodiment may comprise polyoxyethylene) and
poly(oxypropylene), polyethylene glycol esters of fatty acids, e.g.
coconut, polysorbate, polyoxyethylene or polyoxypropylene ethers of
higher alkanes (C12-C18). Examples include Tween.RTM. 20
(polysorbate 20) and Tween.RTM. 80, polyoxyethylene (23) lauryl
ether (Brij.RTM. 35), polyoxyethyene (40) stearate (Myrj.RTM. 52),
polyoxyethylene (25) propylene glycol stearate (Atlas.RTM. G
2612).
[0590] An amphoteric, cationic or anionic surfactant may also be
present in the contact lens solution. Amphoteric surfactants
suitable for use in a composition according to the present
invention include materials of the type are offered commercially
under the trade name "Miranol". Another useful class of amphoteric
surfactants are exemplified by cocoamidopropyl betaine commercially
available under the trade name Amphoso CA. Surfactants suitable for
use in the invention can be readily ascertained, in view of the
foregoing description, from McCutcheon's Detergents and
Emulsifiers, North American Edition, McCutcheon Division, MC
Publishing Co., Glen Rock, N.J. 07452 and the CTFA International
Cosmetic Ingredient Handbook, Published by The Cosmetic, Toiletry,
and Fragrance Association, Washington, D.C.
[0591] The pH of some contact lens solutions should, in one
embodiment, be maintained within the range of about 6.0 to 8.0,
preferably about 6.5 to 7.8. Suitable buffers may be added, such as
boric acid, sodium borate, potassium citrate, citric acid, sodium
bicarbonate, TRIS and various mixed phosphate buffers (including
combinations of Na.sub.2HPO.sub.4, NaH.sub.2PO.sub.4 and
KH.sub.2PO.sub.4) and mixtures thereof. Generally, buffers will be
used in amounts ranging from about 0.05 to 2.5 percent by weight,
and preferably, from 0.1 to 1.5 percent. In one embodiment, the
contact lens solutions of this invention contain a borate buffer,
comprising one or more of boric acid, sodium borate, potassium
tetraborate, potassium metaborate or mixtures of the same. Also,
various buffer systems such as citrate, phosphate (appropriate
mixtures of Na.sub.2HPO4, NaH.sub.2PO4, and KH.sub.2PO4),
bicarbonate, tromethamine and other appropriate nitrogen-containing
buffers (such as ACES, BES, BICINE, BIS-Tris, BIS-Tris Propane,
HEPES, HEPPS, imidazole, MES, MOPS, PIPES, TAPS, TES, Tricine) can
be used to ensure a physiologic pH between about pH 6.5 and
8.5.
[0592] In one embodiment, the contact lens solutions of the present
invention are also adjusted with tonicity agents, to approximate
the osmotic pressure of normal lacrimal fluids which is equivalent
to a 0.9 percent solution of sodium chloride or 2.5 percent of
glycerol solution. The solutions are made substantially isotonic
with physiological saline used alone or in combination. In another
embodiment, propylene glycol, lactulose, trehalose, sorbitol,
mannitol or other osmotic agents may also be added to replace some
or all of the sodium chloride.
[0593] Examples of suitable tonicity-adjusting agents include, but
are not limited to: sodium and potassium chloride, dextrose,
glycerin, calcium and magnesium chloride. These agents are
typically used individually in amounts ranging from about 0.01 to
2.5% (w/v) and preferably, from about 0.2 to about 1.5% (w/v).
Preferably, the tonicity agent will be employed in an amount to
provide a final osmotic value of 200 to 400 mOsm/kg, and more
preferably between about 250 to about 350 mOsm/kg, and most
preferably between about 280 to about 320 mOsm/kg.
[0594] It may also be desirable to optionally include water-soluble
viscosity builders in the solutions of the present invention.
Because of their demulcent effect, viscosity builders have a
tendency to further enhance a lens wearer's comfort by means of a
film on the lens surface cushioning impact against the eye.
Included among the water-soluble viscosity builders are polymers
like polyvinylalcohol cellulose-derived polymers, and povidone. In
another embodiment, viscosity builders comprise polyethylene
glycol, surfactants, polyvinylpyrrolidone, polyvinyl alcohol,
carboxymethyl cellulose and similar materials. Such polymers may be
used in an amount of from about 0.01 to about 4.0 weight percent or
less. Surface-active agents, such as polysorbates, polyoxyethylenes
and certain phosphonates, may be added to ensure proper wetting
and/or cleaning. Sequestering agents such as
ethylenedianiinetetraacetic acid (EDTA), phosphonates, citrate,
gluconate and tartarate, are also common additives to
preservatives, disinfection or cleaning solutions.
[0595] In another embodiment, nitrogen (non-fluorocarbon) is used
as a propellant in the contact lens solution.
[0596] In another embodiment, the contact lens solution of the
instant invention further comprises anti-microbial compounds, which
in one embodiment, comprise glycosides, alkaloids, phenolics
(anthocyanins, quinones, flavonols and flavonoids, etc.),
terpenoids (including phytosterols and carotenoids), or a
combination thereof. In another embodiment, anti-microbial
compounds comprise allicin, aucubin, berberine, bilberry extract,
caffeic acid, chlorogenic acid, Echinacea extract, ferulic acid,
hydrastine, lipoic acid, naringin, oleuropein, proanthocyanidins,
quercetin, rutin, or a combination thereof, which in one embodiment
are present in amounts of 10 to 10,000 parts per million. In
another embodiment, saponins, can be used as natural plant
surface-active or cleaning agents in lens solutions. Specifically,
triterpenoid saponins and steroid saponins are particularly
effective in contact lens or ophthalmic solutions. In another
embodiment, the solution comprises benzyldimethyl
{2-[2-(p-1,1,3,3-tetramethylbutylphenoxy)ethoxy]ethyl}ammonium
chloride (BDT) as an anti-microbial compound.
[0597] In another embodiment, a contact lens solution of the
instant invention comprises preservatives, which in one embodiment
comprise thimerosal, edetate disodium, sorbic acid, polyaminopropyl
biguanide, POLYQUAD (polyquartenium-1), EDTA, or a combination
thereof.
[0598] In one embodiment, a contact lens solution may be a daily
cleaner, which in one embodiment comprises
cocoamphocarboxyglycinate, sodium lauryl sulfate, hexylene glycol,
sodium chloride, sodium, Tween 21, microlens poloxamer 407,
potassium chloride, poloxamine, isopropyl alcohol, amphoteric 10,
or a combination thereof.
[0599] In one embodiment, contact lens solution may comprise an
enzyme, which in one embodiment may be pancreatin, papain,
subtilisin, or a combination thereof, which in one embodiment may
be for removing protein from the lens.
[0600] In another embodiment, a device for use with contact lenses
may comprise compounds for use in the instant invention. In one
embodiment, such a device may be a standard device commercially
available for the storage, cleaning, disinfection, and/or carriage
of contact lenses, which in one embodiment, is characterized by a
hollow well. In one embodiment, a single device may be suitable for
storage, cleaning, disinfection, and/or carriage of contact lenses,
while in another embodiment, each is a separate device.
[0601] In one embodiment, the substrate, which in one embodiment is
a contact lens, contact lens device, contact lens solution and/or
intraocular device, comprising compounds for use in the instant
invention suppresses, inhibits, prevents or treats eye-related
disorders, including, inter alia, those described hereinabove, in a
subject. In another embodiment, the substrate comprising compounds
for use in the instant invention prevents or treats proteinaceous
deposits accumulating on the substrate. In another embodiment, the
compounds for use in the instant invention endow the surface of the
substrate with the property of being more hydrophilic, which in one
embodiment, may increase comfort, decrease eye dryness, or a
combination thereof. In another embodiment, the compounds for use
in the instant invention prevent adverse reactions that are
directly or indirectly related to the substrate, such as corneal
edema, inflammation, or lymphocyte infiltration. In another
embodiment, the substrate comprising compounds for use in the
instant invention increases wettability, decreases adhesion,
increases biocompatability, provides UV shielding, prevents glare,
decreases dryness, grittiness, general discomfort, prevents
microbial (in one embodiment, bacterial) infections, or a
combination thereof or provides other desirable characteristics and
properties to the substrate that are known in the Art.
Preparation of Compounds for Use in the Present Invention
[0602] In one embodiment, the preparation of high molecular weight
compounds for use in the methods of the present invention is as
described in U.S. Pat. No. 5,064,817, which is incorporated fully
herein by reference. In one embodiment, these synthetic methods are
applicable to the preparation of low molecular weight compounds for
use in the present invention as well, i.e. compounds for use in the
present invention comprising monomers and dimers as the conjugated
moiety, with appropriate modifications in the procedure as would be
readily evident to one skilled in the art. The preparation of some
low molecular weight compounds for use in the present invention may
be conducted using methods well known in the art or as described in
U.S. patent application Ser. No. 10/952,496, which is incorporated
herein by reference in its entirety.
[0603] Without further elaboration, it is believed that one skilled
in the art can, using the preceding description, utilize the
present invention to its fullest extent. The following preferred
specific embodiments are, therefore, to be construed as merely
illustrative, and not limiting the remainder of the disclosure in
any way whatsoever.
EXAMPLES
[0604] The abbreviations used in the examples below are:
[0605] PE=phosphatidyl-ethanolamine
[0606] HA=hyaluronic acid
[0607] Cpd=Compound
[0608] Cpd XXII=dipalmitoyl-PE conjugated to HA
[0609] Cpd XXIII=dimyristoyl-phosphatidyl-ethanolamine linked to
HA
[0610] Cpd XXIV=PE conjugated to heparin
[0611] Cpd XXV=PE conjugated to chondroitin sulfate A (CSA)
[0612] Cpd XXVI=PE conjugated to carboxymethyl cellulose (CMC)
[0613] Cpd XXVII=PE conjugated to Polygeline (haemaccel)
[0614] Cpd XXIX=PE conjugated to dextran
[0615] Cpd XXX=PE conjugated to aspirin
[0616] Cpd LXXXVIII=PE conjugated to glutaryl
[0617] Cpd LI=PE conjugated to alginate
[0618] The compounds used in the examples below were prepared as
described in U.S. patent application Ser. No. 10/952,496, which is
fully incorporated herein by reference.
Example 1
Effect of Lipid Conjugates in an In Vitro Model of Diabetic
Retinopathy
Human Retinal Endothelial Cell Culture
[0619] Human eyes from donors are obtained and human retinal
endothelial cells (HRECs) are isolated. The identity of HRECs is
validated by demonstrating endothelial cell incorporation of
fluorescence-labeled, acetylated LDL, and by fluorescence-activated
cell-sorting analysis. To determine the effect of high glucose,
HRECs are grown for 7 days in normal (5.5 mM) or high (25 mM)
D-glucose medium.
[0620] Bovine Retinal Endothelial Cell Culture
[0621] Isolated bovine retinas are homogenized in ice-cold Eagle's
minimal essential medium (MEM) with HEPES by a Teflon-glass
homogenizer and microvessels trapped on an 83 mm nylon mesh.
Vessels are transferred into 2.times. MEM containing 500 .mu.g/ml
collagenase, 200 .mu.g/ml pronase (BDH, Poole, UK) and 200 .mu.g/ml
DNase at 37.degree. C. for 20 min The resultant vessel fragments
are trapped on 53 .mu.m mesh, washed with cold MEM, and centrifuged
at 225.times.g for 10 min. The pellet is suspended in microvascular
endothelial cell basal medium (MECBM) with growth supplement (TCS
Works Ltd., Buckingham, UK) at 37.degree. C., 5% CO.sub.2 for 3
days. Confluent cells are used between passages 1 and 3.
[0622] ELISA for VEGF
[0623] VEGF protein concentration is determined from retinal
endothelial cell culture-conditioned medium using the
Quantikine.RTM. Human VEGF Immunoassay ELISA kit (R & D
Systems). Retinal endothelial cell culture are treated with either
5.5 or 25 mM glucose and aliquots are taken daily for analysis.
[0624] ELISA for IGF-I
[0625] IGF-I protein concentration is determined from retinal
endothelial cell culture-conditioned medium using the
Quantikine.RTM. Human IGF Immunoassay ELISA kit (R & D
Systems). Retinal endothelial cell culture are treated with either
5.5 or 25 mM glucose and aliquots are taken daily for analysis.
[0626] Statistical Analysis
[0627] Data are analyzed using the Student's t-test and reported as
mean.+-.standard deviation (SD). A p value <0.05 is considered
significant.
[0628] Treatment with 1, 5, and 10 .mu.M, and other concentrations
of Compounds XXII, XXIII, or XXV restore the levels of IGF-1 and
VEGF in human and bovine retinal endothelial cell culture to the
level of controls.
[0629] In another embodiment, other markers may be examined
including ICAM-1, VCAM-1, HIF-1, transmembrane reductase (TMR), and
EPO; pigment epithelium-derived factor (PEDF) in the eye, markers
of oxidative stress including osmotic stress after accumulation of
sorbitol, increased cytosolic NADH/NAD ratio, depletion of NADPH
and accumulation of fructose with the resulting non-enzymatic
production of advanced glycation end products (AGES); and/or ESR,
fibrinogen, SDF/1_, RANTES, EpOx, Haptoglobin and ACE in peripheral
blood. Antibodies are used to probe the GAG portion of the
conjugate over a time course compared to unconjugated control, and
show greater local persistence. In one embodiment, GAGs are
tagged.
Example 2
Effect of Lipid Conjugates in an In Vivo Model of Diabetic
Retinopathy
[0630] Diabetes is induced Long-Evans rats via ip streptozotocin
(STZ) injections at 70-85 mg/kg, for 3 to 5 days. To help ease the
transition to diabetes, the rats are given 10% sugar water for 24
hours post-STZ injection. Retinal photographs are taken and blood
glucose tests are performed to determine baselines for each rat.
The normal glucose range for a rat is 80-100 mg/dl.
[0631] Alternatively, C57BL/6 mice (SLC, Shizuoka, Japan) are used.
Postnatal day (P)7 mice with their nursing mothers are maintained
for a full 5 days in 80% oxygen to generate the nonvascular retinal
area. On P12, they are placed in normoxia for an additional 5 days
to induce retinal neovascularization.
[0632] Each animal is glucose-tested and photographed with a fundus
camera on a weekly basis to record the progression of diabetic
retinopathy. Approximately 20-30 minutes prior to starting
pictures, one drop of 1% atropine is placed in each eye. The rats
are anesthetized with sodium pentobarbital, at a dose of 60 mg/kg,
to keep them immobile, and then injected ip with 0.1 ml of 25%
fluorescein, which is used to visualize the retinal blood vessels
when illuminated by blue light, and to determine the relative
leakage of blood by the intensity in the background. The retinal
leakage score is determined by digital analysis of the vascular and
extravascular fluorescence.
[0633] Rats treated with PTZ show significantly increased retinal
leakage scores compared to vehicle-treated controls. Treatment with
1, 5, and 10 .mu.M or other concentrations of Compounds XXII,
XXIII, or XXV decrease the retinal leakage score of PTZ-treated
rats back to the level of controls.
[0634] After 14 days, rats are sacrificed and their retinas
examined for retinal mRNA and protein levels of intercellular
adhesion molecule (ICAM)-1, vascular endothelial growth factor
(VEGF) by RT-PCR and ELISA.
[0635] RT-PCR for Intercellular Adhesion Molecule-1 and Vascular
Endothelial Growth Factor Receptor-1 and -2
[0636] Total RNA is isolated from the retina using extraction
reagent (Isogen; Nippon Gene, Toyama, Japan) and
reverse-transcribed with a cDNA synthesis kit (First-Strand;
Pharmacia Biotech, Uppsala, Sweden) according to the manufacturer's
protocols. PCR is performed with Taq DNA polymerase (Toyobo, Tokyo,
Japan) in a thermal controller (MiniCycler; MJ Research, Watertown,
Mass.). The primer sequences are as follows: 5'-ATG TGG CAC CAC ACC
TTC TAC AAT GAG CTG CG-3' (sense) and 5'-CGT CAT ACT CCT GCT TGC
TGA TCC ACA TCT GC-3' (antisense; 37 bp) for B-actin, 5'-GTG TCG
AGC TTT GGG ATG GTA-3' (sense) and 5'-CTG GGC TTG GAG ACT CAG TG-3'
(antisense; 505 bp) for mouse intercellular adhesion molecule
(ICAM)-1. Human/mouse vascular endothelial growth factor receptor-1
(VEGF R1) primers (302 bp; PCR Primer Pair; R&D Systems, Inc.,
Minneapolis, Minn.) and human/mouse VEGF R2 primers (569 bp; PCR
Primer Pair; R&D Systems, Inc.) are used for VEGFR-1 and -2,
respectively.
[0637] ELISA for ICAM-1 and VEGFR-1 and -2
[0638] The animals are killed with an overdose of anesthesia, and
the eyes are immediately enucleated. The retina is carefully
isolated and placed into 200 .mu.L lysis buffer (0.02 M HEPES, 10%
glycerol, 10 mM Na.sub.4P.sub.2O.sub.7, 100 .mu.M Na.sub.3VO.sub.4,
1% Triton, 100 mM NaF, 4 mM EDTA [pH 8.0]) supplemented with
protease inhibitors, and sonicated. The lysate is centrifuged at
15,000 rpm for 15 minutes at 4.degree. C., and the ICAM-1 and
VEGFR-1 and -2 levels in the supernatant are determined with mouse
ICAM-1 and VEGFR-1 and -2 kits (Techne Corp., Minneapolis, Minn.)
according to the manufacturer's protocol. The tissue sample
concentration is calculated from a standard curve and corrected for
protein concentration.
[0639] Treatment with 1, 5, and 10 .mu.M and other concentrations
of Compounds XXII, XXIII, or XXV dose-dependently decreases the
levels of ICAM-1 and VEGF in PTZ-treated rats back to the level of
controls.
Example 3
Effect of Lipid Conjugates in Patients with Diabetic
Retinopathy
[0640] Plasma samples are collected from Type I diabetic patients.
Their grade of retinopathy is characterised according to a modified
Airlie house technique: (a) no retinopathy (n=6), (b) background
retinopathy (n=10), (c) proliferative retinopathy (n=6), and (d)
advanced proliferative retinopathy requiring vitrectomy (n=16).
Plasma samples from non-diabetic age-matched control subjects are
also collected. Vitreous samples are collected from patients with
advanced proliferative diabetic retinopathy prior to undergoing
vitrectomy. Research Ethics Committee approval and informed consent
are obtained from all patients.
[0641] Plasma and Vitreous Samples
[0642] Venous blood samples are collected from the patients and
control subjects. Plasma is harvested by centrifugation, aliquoted
and stored at -70.degree. C. Approximately 0.5-1 ml of undiluted
vitreous fluid is collected from the eye prior to irrigation of the
vitreous, transported on dry ice and stored at -70.degree. C.
[0643] Enzyme-Linked Immunosorbent Assay (ELISA) for CD105
[0644] White 96-well micro-titre plates are coated with anti-CD105
Mab E9 (100 .mu.l/well) diluted 1/1000 in 0.1 M PBS, and incubated
in a humidified chamber overnight at 4.degree. C. The coated plates
are blocked using 1% BSA and 0.1% Tween 20 in 0.1 M PBS (PBS-Tween)
for 2 h at room temperature. Test samples, 1/2 diluted in
PBS-Tween, are added to the plates in duplicate. Plasma with
pre-determined CD105 (100 ng/ml) is titrated to make a standard
curve in each plate. After overnight incubation at 4.degree. C.,
biotinylated Mab E9 (1/2000 dilution), 100 .mu.l/well, is added to
the plates, followed by incubation at 4.degree. C. in a humidified
chamber for 3 h. HRP-conjugated avidin at 1/2000 dilution in
PBS-Tween and 1% BSA is added(100 .mu.l/well), and plates are
incubated at room temperature for 30 min Three washes with
PBS-Tween are carried out between each of the procedures. Finally,
100 .mu.l/well of Amerlite signal reagent (Amersham UK) are added
to each well and light emission is measured immediately at 420 nm
in an Amerlite plate reader (Kodak Clinical Diagnostics, Aylesbury,
UK).
[0645] Indirect Immunoassay for VEGF
[0646] White 96-well plates are coated with 1000/well of goat
anti-VEGF-165 antibody (R&D systems), diluted 1/1000 (1
.mu.g/ml) in 0.1M carbonate buffer (pH 9.6), and incubated in a
humid box overnight at 4.degree. C. The coated plate is blocked
with 1% (w/v) bovine serum albumin (BSA), 0.01% (v/v) Tween 20 in
0.1 M PBS (PBSTween) for 2 h at room temperature. Serum samples are
added in duplicate to the plates (100 .mu.l/well, diluted 1/2 in
PBS-Tween). A standard curve is generated using recombinant human
VEGF (R & D systems) in a range of 0.1-40 ng/ml on each plate.
After overnight incubation at 4.degree. C., rabbit anti-VEGF
antibody (Santa Cruz Biotechnology) is added (100 .mu.l/well) to
the plate at 1/2000 dilution (1 .mu.g/ml) in PBS-Tween and
incubation is carried out for 3 h at 4.degree. C. This is followed
by the addition of HRP-conjugated goat anti-rabbit antibody
(0.5.mu.g/ml) (diluted 1/2000 with 1%BSA in PBSTween), and
additional incubation with shaking for 30 min at room temperature.
Three washes with PBS-Tween are carried out between each of the
steps. Finally, 100 .mu.l/well of Amerlite chemiluminescence signal
reagent are added and the plate is read immediately in a plate
reader. The measured values of light emission are converted into
absolute concentration by reference to the VEGF standard curve.
[0647] Treatment with 1, 5, and 10 .mu.M and other concentrations
of Compounds XXII, XXIII, or XXV dose-dependently decreases the
plasma levels of CD105 and vitreous levels of VEGF in patients with
diabetic retinopathy. In addition, fluorescein angiography, retinal
photography, and ultrasound imaging of the eye are used to evaluate
progression of the disease.
Example 4
Effect of Contact Lens Solution Comprising Lipid Conjugates on
Protein and Lipid Deposition
[0648] Contact lenses are exposed to both a protein and lipid
artificial deposition solution (ATS) in order to assess both the
deposit inhibition of the contact lens solution of the instant
invention compared to a solution known in the Art, such as
ReNu.RTM. Rewetting Drops which contains 0.10% poloxamine, 0.50%
boric acid, 0.35% sodium borate, 0.40% sodium chloride, 0.10% EDTA,
and 0.15% sorbic acid. To test for deposit inhibition, lenses are
preconditioned with the solution of the instant invention by
soaking the lens in the solution for one hour prior to deposition.
After deposition and incubation, the lenses are rinsed with 0.9%
saline solution (without sorbic acid).
[0649] A. Protocol for Testing Protein Deposit Inhibition:
[0650] For preparation of the standards, unworn contact lenses are
taken out of their vials, left to air-dry and then placed in glass
test tubes along with standard BSA solution. An in vitro protein
mixture consisting of lysozyme, lactoferrin, human serum albumin
and mucin in MOPS buffer is used. The pH of the solution is
adjusted to 7.2 using 1 N HCl and an osmolality equal to 326 mOsm.
After one hour of pre-soaking, the lenses are removed from the
formulation and placed in 1.5 ml of the protein mix. The lenses are
then incubated in the protein mix at 37.degree. C. in a shaking
water bath for 48 hours. Protein analysis is done using the
colorimetric BCA analytical method (Sigma). The method employs the
protein induced reduction of Cu(II) to Cu(I). A purple complex
(Amax=562 nm) is formed following the addition of bicinchoninic
acid (BCA) to the reduced copper. The intensity of the complex is
directly proportional over the protein concentration range of 5
.mu.g/ml to 2000 .mu.g/ml. Following incubation at 37.degree. C.,
the rate of color development is slowed sufficiently to allow large
numbers of samples to be assayed in a single run. The standard
protein solution utilized is BSA with a standard concentration
range of 0 to 50 .mu.g. Two mls of a mixture of bicinchoninic acid
(BCA) and Cu(II) sulfate is added to each test tube, which are then
vortexed. Tubes are then covered and placed in a water bath at
37.degree. C. for 15 minutes. After incubation, the purple complex
develops. Samples and standards are read in a spectrophotometer at
562 nm Protein concentration is determined from a standard plot of
absorbency vs. concentration (ig).
[0651] B. Protocol for Testing Lipid Deposit Inhibition:
[0652] Seven contact lenses per test solution are preconditioned
with the respective test formulations by soaking the lenses in the
formulation for one hour. The lenses are removed from the
formulation, and placed in 1.5 mls of a lipid mix (palmitic acid
methyl ester (PAME), cholesterol, squalene and mucin in MOPS
buffer). Mucin is utilized as a surfactant to aid in the
solubilization of the lipids. Lenses are then incubated in the
lipid mix at 37.degree. C. in a shaking water bath for 24 hours.
After incubation, the lenses are removed from the test solution and
rinsed with physiological saline solution (without sorbic acid) to
remove any residual deposition solution. Lenses are then placed in
glass vials for extraction. A three-hour 1:1 CHCl.sub.3/MeOH
extraction is subsequently followed by a three-hour hexane
extraction. Extracts are then combined and run on a Hewlett Packard
GC. The column utilized is an HP-Ultra 1 with an FID detector and
He as the carrier gas. Standard solutions of each of the lipids in
the deposition mix are made in 1:1 CHCl.sub.3/MeOH and the
concentration of lipid extracted from the lenses is determined
[0653] C. Results:
[0654] The protein and lipid deposition values for the contact
lenses pre-soaked in control solution provide a baseline with which
to assess the potential cleaning efficacy and deposit inhibition
attributes of each of the formulations tested. The contact lens
solution of the instant invention inhibits both lipid and protein
deposition, indicating that the test formulations are coating the
lens in such a way as to hinder lipid and protein uptake.
Example 5
Safety and Tolerability of Contact Lens Solution Comprising Lipid
Conjugates
[0655] Twenty (20) subjects are enrolled in a 4-hour non-dispensing
study comparing the contact lens solution comprising
lipid-conjugates to contact lens solution comprising unconjugated
GAGs or to ReNu.RTM. Rewetting Drops. The subjects are all habitual
soft spherical contact lens wearers. Their mean spherical Rx's are
determined for test and control eyes. Each subject wears a pair of
contact lenses for approximately 4 hours. The eye receiving the
test solution is randomly selected and remains constant for the
duration of the study. Subjects are asked to place two drops of
each solution into the appropriate eye every hour until the
four-hour visit. The subjects and investigator are blinded to
solution identity. Prior to lens insertion, a spherical refraction
is performed through which high contrast visual acuity with high
ambient illumination (HCHI) is measured. Corneal and conjunctival
staining and limbal and bulbar injection are assessed with the
slitlamp. Each subject is then fitted with a pair of contact lenses
of their prescription. Each lens is evaluated for centration and
movement, comfort, and deposits/wettability. A spherical
over-refraction is then performed. The endpoint of the
over-refraction is compared to the refractive endpoint to determine
the apparent "on-eye" lens power. LogMAR visual acuity under HCHI
testing conditions is measured through the over-refraction.
Finally, two drops of each solution are instilled into the
appropriate eyes, and the subject is asked to rate any sting/burn
and the amount. Testing is repeated at the four-hour visit in
reverse order, except without repeating the baseline refraction. A
two-way ANOVA incorporating Time and Solution is used to test for
differences in each of the parametric dependent variables measured.
Non-parametric data are analyzed by Friedman ANOVA. Differences at
the p<=0.05 level are considered to be statistically
significant.
[0656] Subjects are evaluated for the effects of the solution
comprising lipid-conjugates for comfort, apparent lens Rx power,
and sting/burn visual analog score (i.e. lower sting/burn), lens
movement/centration, and anterior ocular physiology in the eye
treated with lipid-conjugate solutions compared to the eye treated
with control solution.
Example 6
Safety and Tolerability of Contact Lens Packaging Solution
Comprising Lipid Conjugates Materials and Methods
[0657] Contact lenses were exposed to a solution comprising lipid
conjugates in order to assess their efficacy as a comfort
ingredient in a contact lens solution. Five lipid conjugate
compounds, Cpd LI (AlgPE:120), Cpd XXV (CSAPE:120), Cpd XL
(HemPE:75), Cpd XXVIII (HesDMPE:90 (HetaStarch)) and Cpd XXVI
(CMPE:75), were tested.
[0658] Cpd LI, Cpd XXV, Cpd XL, Cpd XXVIII and Cpd XXVI were tested
for their solubility in an aqueous solution containing
Na.sub.2HPO.sub.4.times.7H.sub.2O, NaH.sub.2PO.sub.4 and NaCl, pH
7.3, with an osmolality of 250 mOsm/kg.
[0659] PV lenses were tested for lens compatibility and stability.
Each lens was soaked in a glass vial containing 3 ml of a solution
of one of the following: Cpd XXV, Cpd XL, Cpd XXVIII or Cpd XXVI.
The glass vials were autoclaved at 121-123.degree. C. for 30 min
The lenses and the solutions were examined for optical parameters
and physical appearance immediately following autoclaving and again
after one month at 40.degree. C.
[0660] Cytotoxic effects of the compounds XXV, XL, XXVIII and XXVI
on kidney (MDCK) and SV40 human corneal (HCEC) epithelial cells
were evaluated. Each of the compounds in solution was added to MDCK
and HCEC cell culture models, with physiological saline or HBSS
serving as a control. The compounds were individually tested for
their toxicity to L929 mouse fibroblasts in an agar diffusion
model, in which cells were separated from each compound by a layer
of agar. In this test only compounds that can diffuse through the
agar and are toxic to the cells are detected. The L929 monolayer
cultures were incubated with the compounds, and observed for
cytotoxicity.
[0661] PV lenses were soaked in solutions of the compounds and
tested. In vivo evaluation was assessed in a one-day rabbit ocular
irritation experiment. PV lenses were soaked in solutions of the
compounds and placed on the corneas of test rabbits.
Results
[0662] Cpds XXV, XL and XXVIII were soluble. Cpd XXVI was soluble
upon autoclaving the solution. Cpd LI was insoluble.
[0663] The optical parameters and physical appearance of the
contact lenses immediately following autoclaving were within
industry specifications. There were no significant changes in the
pH or osmolality of any of the solutions. A follow-up check one
month later also revealed no significant changes in either lens or
solution parameters.
[0664] Results with the compounds on MDCK and HCEC paralleled those
of the controls. No cytotoxicity to L929 monolayer cultures was
noted.
[0665] No or low levels of ocular irritation were observed with the
PV lenses in vivo. There was no statistical difference between
treated vs. control PV lenses.
[0666] It will be appreciated by persons skilled in the art that
the present invention is not limited by what has been particularly
shown and described herein above and that numerous modifications,
all of which fall within the scope of the present invention, exist.
Rather, the scope of the invention is defined by the claims which
follow:
Sequence CWU 1
1
4132DNAArtificial SequencePrimer 1atgtggcacc acaccttcta caatgagctg
cg 32232DNAArtificial SequencePrimer 2cgtcatactc ctgcttgctg
atccacatct gc 32321DNAArtificial SequencePrimer 3gtgtcgagct
ttgggatggt a 21420DNAArtificial SequencePrimer 4ctgggcttgg
agactcagtg 20
* * * * *