FOR REDUCING LEVELS OF ARACHIDONIC ACID I TISSUE HAVING UNDERGONE AN INVASIVE PROCEDURE

I■ The Field of the Invention

This invention relates to compositions and methods for reducing levels of araehidonic acid in tissue after surgery. In particular, the present invention is directed to compositions- for reducing levels of araehidonic acid in tissue having, undergone an invasive -procedure., wherein the composition comprises at least one fatty acid. Correspondingly, the present invention is further directed to methods for reducin levels of araehidonic acid in tissue after surgery and, accordingly, reducing the resul ting tissue inflammation, comprising the steps of admmistering preoperatively and/or postoperatively a composition comprising at least one fatty acid, wherein the fatty acid may compri se omega-3 fatty acids; increasing levels of anti-inflammatory omegas' s in said tissue; and decreasing levels of inflammatory omega-6's in said tissue. In certain embodiments, the omega-3 fatty acids comprise- the triglyceride form delivered in an effective daily dosage. 2- The Background Art.

Dry eye is a. conditio in which there are insufficient tears to lubricate and nourish the eye. Tears are necessary for maintaining the health of the front surface of the eye and for providing clear vision. People with dry eyes either do not produce enough tears or have a poor quality of tears. With each blink of the eyelids, tears are spread across the cornea in order to provide lubrication, to wash away any foreign matter and to keep the surface of the eyes smooth and clear.

Tears are produced by several glands in and around the eyelids. When the normal amount of tear production decreases or tears evaporate too quickly from the corneal surface, symptoms of dry eye can develop.

As appreciated, tears are made up of oil, water and mucus. Each component serves a specified function in protecting and nourishing the front -surface of the eye. A smooth oil layer helps to prevent evaporation of the water layer, while the mucin layer functions in spreading the tears evenly over the surface of the eye. If the tears evaporate too quickly or do not spread evenly over the cornea as a result of deficiencies with any of the three tear layers,, symptoms of dry eye or posterior blepharitis may ensue. Along the margin of the ^' eyelids are a series of small ^' sebaceous glands, called meibomian glands. The ^' meibomian glands creat and distribute a supply ©f meibum., an oily substance, that makes up the lipid layer of the tear. The supply ofmeibum functions to help keep the eye moist and tends to protect the tear film, from evaporation. There are approximately twenty-five meibomian glands on the upper eyelids and twenty-fi ve meibomian glands on the lo wer eyelids. Upon blinking of the eye. the upper eyelid comes down, presses on the oily substance produced by the meibomian glands, and pulls a sheet of this oily substance upwards, thereby coating the tear layer beneath to keep it from evaporating. This oily substance or meibum (wherein lipids are axnajor component) which is created by the meibomian glands is therefore critical for healthy eyes and clear vision.

Meibomianitis refers to inflammation or dysfunction of the meibomian glands which is also referred to in the art as meibomian gland dysfunction. Inflammation of the meibomian, glands may occur because of the production of meibum which is pro-inflammatory in nature as a result of an increased composition of omega-6 essential fatly acids. Secondarily, bacteria have been found to invade the meibomian glands and colonize there, Once inflamed, the meibomian glands generally will not function in a manner sufficient to adequately produce the quantity and quality of oils necessary to properly lubricate the eye.

The volume of oil produced from inflamed meibomian glands tends to decrease and the oils that are produced become thicker in composition, like toothpaste. These oils also become abnormal in their characteristics. Instead of spreading evenly across the aqueous layer, the oil coalesces ^' lea ving areas on the corneal surface in which the aqueou can evaporate and other areas in which the oil adheres to the cornea surface itself. This creates a dry spot on the cornea for which the aqueous cannot penetrate. Such condition generally produces a foreign body sensation and if it persists may result in injury to the epithelium which is seen as corneal Staining on examination. A reduction in oil production therefore inherently results in a quantitative decrease in the quality and quantity of the oily layer, thus causing tears to evaporate more rapidly. Because the thickened oil does not coat the eye properly, a person with inflamed meibomian glands may experience discomfort or problems with, their eyes that may include, for example, but not by way of limitation: (1) dryness; (2) burning; (3) itching; (4) irritation and redness: (5) blurred vision: and/or (6) foreign body sensations,

Tills inflammatory process can also spread throughout the lid margin and spill over to involve the ocular surface resulting in significant ocular discomfort.. Inflammation of the meibomian glands in the upper and lower lids can further lead to vascularization and fibrosis, causing stenosis and then closure of the meibomian gland orifices. Deprived of the meibum or lipids that inhibit evaporation, tear film evaporation will generally increase. Similarly, a deficiency in tea film generally results in irritation of the eye, but can also cause damage to the surface of the eye. As appreciated, an irregular oil pattern disrupts tears and allows for increased exposure of the aqueous layer to the atmosphere and the increased evaporation of the aqueous. Unfortunately, this inflamed condition of the meibomian glands has often been found to be chronic.

Some of the treatments ^' for meibomianitis that have been contemplated by those skilled in the art incl ude: (J) the application of artificial tears; (2) cleaning the affected eyelid margins with a gentle baby shampoo; and (3) applying warm and moist compresses 5-10 minutes two to three times per day in an effort to promote normal eyelid glandular function. A physician may also prescribe a topical and/or oral antibiotic such as, for example, tetracycline, erythromycin, or doxyeycline, to help eradicate the bacteria found in the glands and to facilitate a breakdown in the thickened lipid secretions from the meibomian glands. These various treatments, however, can often take months before a treated patient notices any significant improvement.

Although the elimination of bacteria or antiinflammatory effects of the antibiotics resulted, in a temporary change, none of the known treatment methodologies have brought long- lasting relief to patients. Hoping to provide a form of sustainable relief to the ongoing symptoms associated with dry eye, with or without meibomian gland dysfunction, that are suffered by patients, a study was conducted by those skilled In the art to investigate the effects of dietary supplementation of a combination of flaxseed and fish oils on the teax film and the ocular surface. At the baseline, all patients in the study had a history of dry eye or one or more symptoms of posterior blepharitis. At the end of the study, the clinical results did not achieve any statistical significance, wherein the lipid composition of the samples collected from the omega-3 supplemented group was found to be very similar to that collected from the placebo group. Thus, the study concluded that dietary supplementation of flaxseed oil and omega-3 fatty acids for treating dry eye or meibomianitis showed no significant effect on meibum composition or aqueous tear evaporation rate.

Consistent with the foregoing, in order to control or resolve the long term effects of dry eye, posterior blepharitis, or meibomian gland dysfunction, the characteristics or nature of the oil (meibum) that is produced by the meibomian glands must be normalised. Thus, what is needed are nutritional or dietary supplement compositions and treatment methodologies using the same that effectively change the quality of the meibum composition,, thereby resulting in a meibum composition having a direct correlat on to enhancing and improving the function .and/or composition of the lipid layer of the tear which reduces the symptoms associated with dry eye, posterior blepharitis and/or meibomian gland dysfunction.

As appreciated, invasive surgical operations may induce an acute inflammatory response thai usually impact the clinical outcomes. Surgical injury induces a systemic inflammatory response proportional to the severity of the insult. An appropriate response maintains homeostasis and allo ws wound healing while an excess! ve response may trigger an inflammatory cascade resulting in the systemic inflammatory response syndrome (SIRS). Correspondingly, tissue injury generally results in cytokine release, which in turn stimulates the production of acute phase proteins such as€ -reactive protein (CRP), fibrinogen, complement G3 and haptoglobin.

For example, when cataract surgery is performed on a patient, the resultant, tissue trauma usually incites inflammation which can cascade leading to not only to pain and discomfort for the patient, but to more serious problems such as cystoid macular edema. Those skilled in this art have developed means to reduce potentially vision-threatening problems from, occurring by means of topically applying a combination of NSAlDs and steroids, which has been shown to reduce tissue inflammation. However, careful monitoring must be undertaken in order to sufficiently devise a treatment duration to avoid and guard against potential complications associated with use of these drugs.

Although the combination of NSAfDs and steroids are topically applied to the eye, it is important to remember regarding inflammation is what transpires on the cellular level concerning additional inflammatory mediators, such as prostaglandins, interleukins, leukotrienes, TNF-a and TGF-β. As appreciated to those skilled in the ait, these molecules in turn, trigger a cascade of intracellular and extracellular changes, including the breakdown of intercellular junctions, which increases vascular permeability.

One particularly significant and worrisome inflammatory cascade is the arachidonic acid, pathway. Surgical trauma results in. the breakdown of cell membrane phospholipids, which phosplvolipase A2 converts into arachidonic aeid. in general,, this arachidonic acid is converted into prostaglandins and leukotrienes, which are both inflammatory mediators. The resulting side effects of these leukotrienes and prostaglandins are pain, infianmiation, macular edema and possibly increased intraocular pressure. The use of a combination of NSAiDs and steroids as physicians are often taught to use should do so with caution when managing postoperative iiiflanimatioa because of the risk of complications .such as cataract, steroid dependence, exacerbation of ^" viral or fungal infections, and intraocular pressure (IOP) elevation. As a result of these complications, a patient's treatment must often be discontinued prematurely or at least reduced in dosage or concentration to diminish the associated inherent risks. At times, steroid treatment is avoided altogether, consequently treating a condition subopiimaily that may recur o become chronic.

Consistent with the foregoing, in order to reduce and manage resulting inflamination associated with tissue after having undergone an invasive surgical procedure, the amount of arachidonie acid in the tissue must be reduced. Thus, what is needed are compositions and methods using the same that not only block the cyciooxygenase enzyme so that arachidonie acid is not converted to prostaglandins and leukotrienes, but replacing the substrate (arachidonie acid) with an antiinflammatory .substance would further reduce overall inflammation,

As contemplated herein, the composition and methods of the present invention comprise the steps of (.1) administering .preoperatively a composition consisting, of at least one fatty acid, wherein the tatty acid may include of omega-3 fatty acids, (2) increasing levels of anti- Inflammatory omega-3 \s in the tissue and (3) decreasing levels of inflammatory omega-6's (arachidonie acid) in the tissue, thereby reducing post surgical inflammation by reducing the prostaglandin precursors and increasing the anti-inflammatory and resoivins available at. the surgical site. The methods of the present invention may further include the step of administering the composition of omega-3. fatt acids, as defined herein, postoperatively. As further contemplated herein, the administration of compositions of the present invention both preoperatively and postoperativel may produce preferred outcomes. Consequently, the compositions and .methods of the present invention have been found to significantly reduce postoperative inflammation of tissue having undergone an invasive surgical procedure, thereby resul ting in reduced, post-surgical downtime and hospital stays and, most important improvement in healing outcomes,

SUMMARY OF THE INVENTION

The present invention discloses methods for administering a supplementation of omega-3 fatty acids to a patient suffering from sym toms of dry eye, wherein the supplementation of omega-3 fatty acids Is provided in an effective -amount sufficient to facilitate an increase in the resulting omega-3's content of the treated meibomian glands, acting as an anti-inflammatory, and, respectively, in a decrease in the amount of resulting omega-6's (arachidonic acid), acting as an inflammatory, thereby having an affect on the normalization of the lipid layer of the tear and a corresponding reduction i the associated dry eye symptoms.

Additionally, the present invention discloses methods for administering a supplementation, of omega-3 fatty acids to a patient suffering from symptom of posterior blepharitis, wherein the supplementation of omega-3 fatty acids in the re-esterified triglyceride form is provided in an effective amount sufficient to effectively change the quality of the meibum composition resulting in a meibum composition that improves or increases tear breakup time, .reduces tear osmolality, and elevates the omega-3 index, while eliminating o reducing the relaxed symptoms of posterior blepharitis.

The present invention further discloses methods for administering a sup lementation of omega-3 fatty acids to a patient suffering from symptoms of meibomianitis, wherein the supplementation of omega-3 fatty acids in the re-esterified triglyceride form is provided in an effective amount sufficient to effectively change the quality of the meibum composition resulting in a meibum composition that improves or increases tear breakup time, reduces tear osmolality, and elevates the omega-3 index, while eliminating or reducing the related symptoms of meibomianitis.

The present invention also discloses methods for changing the composition of the oil produced by sebaceous glands found, in the body from, pro-inflammatory o.m.ega-6 t antiinflammatory omega-3, whereby normalizing the oil production of the treated gland by way of administering a supplementation of omega-3 fatty acids as taught by the present invention. in addition, the present invention discloses methods for ^' changing the composition of the oil (meibum) produced by meibomian glands from pro-inflammatory omega-6 to ami- inflammatory omega-3 , whereby normalizing the oil production of the meibomian gland by way of administering a. supplementation of omega-3 fatty .acids as taught by the present invention.

Furthermore, the present invention discloses methods- for treating post inflammation by preoperati vely and/or postoperatively administering an embodiment of the compositions as taught by the present invention, whereby increasing the omega-3 level and decreasing the level of arachidonic acid (omega-6's) within the cell membrane thereby reducing post surgical inflammation by way of reducing- the prostaglandin precursors and increasing the- antiinflammatory and reso ns available at the surgical site. Consequently, the compositions and methods of the present invention have been found to significantly reduce postoperative inflammation of the tissue having undergone the invasive procedure,^ reduction in post-surgical downtime and hospital stays and. importantly, an improvement in healing outcomes.

Consistent with the foregoing, the present invention, is directed to methods .for administering, a supplementation ofomega-3 fatty acids to a patient suffering from symptoms- of dry eye, posterior blephariti and/or meibomianitis. The supplementation of omega-3 fatty acids is administered in an an¾?unt formulated to- change the composition of the oil (nieibum) produced by meibomian glands from pro-inflammatory omaga-6 to anti-inflammatory omega~3, whereby normalizing the oil production of the meibomian gland so as to improve or increase tear break up- time, reduce tear osmolality, and elevate the omega-3 index, thereby, consequently, eliminating or reducing the related symptoms .of dry eye, posterior blepharitis or meibomian! tis (meibomian gland dysfunction).

In an embodiment of the present invention, the present invention provides for methods for treating and preventing dry eye associated with meibomian gland inflammationor dysfunction by way of -administering a nutritional or dietary supplement composition comprising fin effective amount ofomega-3 fatty acids. The supplementation may include an effective amount ofomega- 3 fatty acids comprising a daily dosage that includes between about 600 mg and about 5,000 n g. The effective amount of omega-3's may comprise the re-esterified triglyceride form.

The effective amount of omega-3 fatty acids may comprise an effective amount of eicosapentaenoic acid (EPA). In one embodiment of the present invention, the daily dosage of an effective amount of EPA may include an amount greater than 600 mg.

in yet another embodiment of the present invention, the effective amount of omega-3 fatty acids may comprise an effective amount of docosahexaenoic acid (DHA). The daily dosage of an effecti ve amount of DHA may include .an amount greater than 500 mg.

in certain embodiments of the present invention, an effective amount of omega-3- fatty acids may be delivered in a daily dosage that includes between about 2,000 mg and about 3,000 mg. This effective amount of omega-3 fatt acids- may comprise a effective amount of eicosapentaenoic acid (EPA) and an effective amount of docosahexaenoic acid (DHA). Similarly, in one embodiment of the present invention, the daily dosage of an effective amount of EPA may include an amount between about 1,600 mg and about 2,500 mg and the daily dosage of an effective amount DHA may include an amount between about 500 mg and about 900 mg. An additional amount of omega~3 fatty acids may also be included in the administered composition. These additional omega-3 fatty acids may include a daily dosage amount of between about 400 mg and about 700 mg. Furthermore, the nutritional or dietary supplement composition of the present invention may include an effective amount, of Vitamin 0 (as 03). An effecti ve amount of Vitamin 0 may comprise a daily dosage amount between about 500 IU and about 2,000 IU.

As further contemplated, the present invention is directed to compositions, for reducing levels of arachidon c acid (omega-6) found in tissue after .surgery, in particular, the present invention involves compositions for reducing levels of arachidonic acid (omega-6) found at the cellular level in tissue having undergone an invasi ve surgical procedure, wherein the eompositi on consists of a single fatty acid.

In certain embodiments of the present invention, the single fatty acid included m the composition consists of omega-3 fatty acids. These omega-3 tatty acids ma comprise the triglyceride form. Additionally, the composition of the present invention may consist of an effective amount of omega-3 fatty acids including between 2,000 mg and 3,000 mg on a daily dosage basis. In certain embodiments of the present invention, the amount of omega-fatty acids found in the composition comprises an -amount greater than 600 mg.

In other embodiments, compositions for reducing levels of arachidonic acid in tissue having undergone an invasive procedure may consist of omega-3 fatty acids comprising an. effective amount of ei-cosapentaenoic acid (EPA). The effective amount of EPA may include an amount between 1,600 mg mid 2,500- mg. In other embodiments of the composition of the present invention, the amount of EPA may be adjusted between. 1,600 mg and 1,800 mg. In further embodiments, the amount of EPA found in the compositions may include about 1,680 The present invention further contemplates compositions for reducing levels of arachidonic acid (omega-6) found at the cellular level in tissue having undergone an invasive procedure, wherein the omega-3 fatty acids of the composition may comprise an effective amount of docosahexaenoic acid (DHA). In certain embodiments, the effective amount of DHA may comprise an amount greater than 500 mg. Alternatively, the effective amount of DHA may comprise an amount between 500 mg and 900 mg. In further embodiments., the amount of DHA found in the composi tions of the present invention may include an amount between 500 mg and 600 mg. In certain embodiments, the compositions of the present invention for reducin levels of arachidonie acid in tissue having undergone an invasive procedure may include omega-3 fatty acids in the triglyceride form. The omega~3 fatty acids contained in the. compositions of the present invention may further comprise a form selected from the group consisting of triglyceride (inclusive of esterified and re-esterified); (2) ethyl ester; (3) free fatty acid; (4) phospholipids; and (5 ) other biochemical forms known in the art.

In some embodiments of the present invention, the composition of omega- 3 fatty acids may include EPA in an amount greate than 600 mg and/or DHA in an amoant greater than 500 mg. In sone embodiments, the composition of the present invention comprises an amount of EPA and an amount of DHA in a 3 : 1 ratio.

Correspondingly, the present invention is further directed to methods for reducing inflammation found in tissue after surgery, in certain embodiments of the present invention, the methods comprise the steps of (1) administering preoperatively a composition consisting of at least one fatty acid, wherein the fatty acid may consists of omega-3 fatty acids, (2) increasing levels of anti-inflammatory omega-3 ⁵ s in the tissue and (3) decreasing levels of inflammatory •omega-6's (arachidonie acid) in the tissue, thereb reducing post surgical inflammation by reducing the prostaglandin precursors, and increasing the antiinflammatory and resolvins available at the surgical site. The methods of the present invention may further Include the step of ^■ administering the composition of omega-3 fatty acids, as defined herein, postoperatively. In certain embodiments, the methods of the present invention contemplate the administration of the compositions taught herein both preoperatively and postoperatively to gamer the synergies associated wit the same.

Consequently, the compositions and methods of the present invention have been found to significantly reduce preoperative inflammation of tissue after an invasive surgical procedure, which, generally results in reduced post-surgical downtime and hospital stays and, most important, improves healing outcomes.

As contemplated herein, the administration of the compositions of the present invention may be delivered by means of softgel, tablet, liquids, granules, .microgranu!es, powders, or any other delivery system deemed effective. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

It will be readily understood that the components of the present invention, as generally described herein, could be modified, arranged and designed in a wide variety of different formula, systems and methods. Thus, the following more detailed description of the embodiments of the composition, systems and methods of the present invention is not intended to limit the scope of the invention. The scope of the in vention is as broad as claimed herein.

As used herein, the term "form" in association with defining the form of omega-3 fatty acids included in the compositions of the present invention means: (!) triglyceride (inclusive of esierified and/or re-esierified); (2) ethyl ester; (3) free fatty acid; (4) phospholipids; and (5) other biochemical forms known in the ait.

As used herein, the term ^' "omega-3 's in the re-esterified triglyceride form" includes omega-3 's derived from marine and other sources. As appreciated, omega-3's in fish are present in the triglyceride form. Marine source fatty acids may undergo purification by the use of absorbents and molecular distillation to remove mercury and other heavy metals and pollutants that are usually prevalent in these sources. This purification process generally results in the omega-3 ¾ being in the ethyl ester form, whichis how the vast majority ofOTC omega-3 products are sold. The omega-3 's derived from marine sources, as used in the studies and as contemplated by the present invention, underwent a further re-esterification step to restore the triglyceride group to the omega-3 's (rTG). Consequently, this further step of re-esterification of the omega- 3's greatly increased the body's ability to absorb the omega-3 's as illustrated in the studies.

As used herein, the term "effective amount" includes the amount of omega-3 fatty acids which is capable of effectively changing the q uality of the meibum concentration which has a direct correlation to improving the lipid layer of the tear, while eliminating or reducing the related symptoms of dry ey e, posterior blepharitis and or meibomianitis.

As used herein, the terms "dry eye, meibomianitis, meibomian gland dysfunction, posterior blepharitis and blepharitis" are to be considered as synonyms.

The present invention provides for methods for treating and preventing dry eye associated with meibomian gland inflammation or dysfunction by way of administering a composition comprising an effective amount of omega-3 fatty acids. The composition may include an effective amount of omega-3 fatty acids comprising a daily dosage that includes between about 600 mg and about 5,000 nig. This effective amount of oniega~3 fatty acids may comprise an effective amount of eicosapentaenoic acid (EPA). In one embodiment of the present invention, the daily dosage of an effective amount of EPA may include an amount greater than 600 mg.

In yet another embodiment of the present invention, the effective amount of omega-3 fatty acids in the composition may comprise an effective amount of docosahexaenoic acid (DMA). The daily dosage of an effective amount of DHA may -include an amount greater than 500 mg.

In certain embodiments _. , the composition of the present invention may include an effective amount of omega-3 fatty acids comprising a daily dosage including an effective amount between about 2,000 mg and about 3,000 mg. This effective amount of omega-3 fatty acids may be comprised of an effective amount of eicosapentaenoic acid (EPA) and an- effective amount of docosahexaenoic acid (DHA). in an embodiment of the present invention, the daily dosage of an effective amount of EPA may include art .amount between about 1,600 mg and about 2,500 mg and the daily dosage of an effective amount DHA may include an amount between about 500 mg and about 900 mg.

As appreciated by those skilled in the art the composition of the present invention includes omega-3 fatty acids that may comprise, for example, but not by way of limitation, the triglyceride form, esterified. and/or re-esterified triglyceride form or concentrates, the ethyl ester form, the free fatty acid form, the phospholipids form, or any other suitable form sufficient to effectively change the quality of the meibum composition of the meibomian glands which has a direct correlation to improving the lipid layer of the tear, while eliminating or reducing the related symptoms of dry eye or meibomianitis. It is contemplated herein mat me supplementation of the omega-3's can he in a concentrated form, whereas up to 100% of the unit, volume can be omega-3. In certain embodiments of the present invention, the dietary or nutritional supplement omega-3 composition administered for treating dry eye, posterior blepharitis, meibomianitis for changing the qualily of the mei bum concentration of inflamed or dysfunctional meibomian glands in order to improve or increase tear break up time, reduce tear osmolality, and elevate the omega-3 index may comprise omega-3 fatty acids in the re-esterified triglyceride form.

The effective amount of eicosapentaenoic acid (EPA) and/or an effective amount of docosahexaenoic acid (DHA) included in the composition of the present invention may be obtained from known sources, such as for example, and not by way of limitation, fish, kri.il, algae, squid, yeast, and vegetable sources, it is further recognized that stearidonic acid is a precursor to EPA and DHA and that consuming a product rich in siearidonic acid may be used to achieve the benefits as disclosed herein.

In selected embodiments of the nutritional or dietary supplement composition of the present invention, an effective amount ofEPA/DHA may be administered in one or more softgel capsules containing an amount in the range of between about 800 -mg and ,250 mg and between about 250 mg and about 450 nig, respectively. For purposes of dosage, in certain embodiments of the present invention, the daily dosage amount ma include an effective amount ofEPA/DHA comprising the amounts of 840 mg and 280 mg, respectively .

in certain embodiments, this effective amount of EPA DHA form ma comprises a ratio ofEPA/DHA of 3: 1. Whereas, in selected embodiments, the ratio of EPA/DHA in each capsule may be in the range of between about 800 mg and i ,250 trig of EPA and between about 250 mg and 450 mg of DHA, whereby two capsules would comprise a daily effective dosage range.

An additional amount of omega-3 fatty acids may also be included in the administered composition. These additional omega-3 fatty acids may include a daily dosage amount of between about 400 mg and about 700 mg.

Fuilliermore, the nutritional or dietary supplement composition of the present invention may include a effective amount of Vitamin D (as D3). Such effective amount of

Vitamin D may comprise a daily dosage amount of betwee about 500 IU and about 2,000 IU.

A clinical study was conducted based on the following parameters:

OBJECTIVE:

To evaluate the clinical ^' effect of the oral administration of a supplementation of omega-3 fatty acids in the re-esterified triglyceride form to a patient suffering from symptoms of dry eye and meibomian gland dysfunction.

SUBJECT ^' S:

A total of twenty-one (21) subjects or participants, between the ages of 18-60 years of age inclusive, who voluntarily provided writte informed consent and who were capable of complying with the study visit schedule, were enrolled. There were three (3) scheduled visits with an attending physician. The first visit included, an initial base line analysis for inclusion in the study. The second visit involved a 4-week follow- up and the third visit was an 8-week follow-up.

STUDY POPULATION:

The parameters of the study protocol for the "inclusion" of participants included the following conditions: (1) the participant must be of the age of 18 to 60 at the time of signing the informed consent; (2) must understand, he willing and able, and likely to fully comply with study procedures, visit schedule, and restrictions; and (3) have symptoms of dry eye, posterior blepharitis, and/or meibomian gland dysfunction.

The parameters of the study protocol for the "exclusion" of participants included the foil owing conditions : ( 1 ) clinically significant eyelid deformity or eyelid movement disorder that is caused by conditions such as notch deformity, incomplete lid closure, entropion, ectropion, hordeola, or chalazia; (2) previous ocular disease leaving sequelae or requiring current topical eye therapy other than fo DEI ⁾ , including, but not limited to: active corneal or conjunctival infection of the eye and ocular surface scarring; (3) active ocular or nasal allergy; (4) LAS1K or PR surgery that was performed within one (1) year of Visit 1 or at any tune during the study; (5) ophthalmologic drop use within 2 hours of Visits 1 , 2, or 3; (6) pregnancy or lactation at any time during the study; (7) abnormal ity of nasolacrimal drainage, (by hi story); (8) prev ious Punctal plugs placement or cauterization; or (9) started or changed the dose of chronic systemic medication known to affect tear production including, but not limited to antihistamines, antidepressants, diuretics, corticosteroids or imniuno-modulators within 30 days of Visit ί , 2, or 3.

STUDY DESIGN:

This is a single-center study of participants with signs and symptoms of dry eye undergoing nutritional therapy treatment with an amount of omega- 3 fatty acids delivered in re- esterified triglyceride form over the course ofthree (3) visits with approximately 4-week intervals between each visit.

The following clinical tests were performed, on each participant at baseline: (!) Ocular- Surface Disease Index (OSDi) which is a survey based on an array of questions that are asked having a gradation scale for answers to score subjective symptoms and to distinguish between normal subjects arid patients with dry eye disease (normal, mild to moderate, and severe) and effect on vision-related function; (2) Slit Lamp Examination which involves the use of a low- power microscope combined with a high-intensity light source that can be focused to shine in a thi beam so that the physician can examine the patient's eyes, especially the eyelids, cornea _* conjunctiva, sclera and iris; (3) Corneal Staining which is an evaluation of epithelial integrity; (4) Tear Break Up Time (TBUT) which involves a method of determining the stability of the tear film and checking for evaporative -dry eye by way o determming the time required for dry spots to appear on the cornea! surface after blinking; (5) Tear Osmolality (Teariab®) that involves measuring the concentration, of the osmotic solution of the tear; (6) EPA and DHA red blood cell saturation using the HS Omega~3 Index (OmegaQuant®) performed by probing the meibomian glands with a Maskin probe tor a meibum sample: and (7) blood omega-3 levels were obtained to ensure patient compliance wit supplementation given.

The participants were placed on a supplementation of omega-3 fatty acids comprising a daily dosage amount of 2,668 rog in are-esterified triglyceride form (rTG) dispensed in four 667 nig capsules, each containing 42.0 mg of EPA, 140 mg of DHA, 107 mg of other omega-3 's and 250 mg ofVitamin D(D3).

The participants were reevaluated at the 4-week visit with all the baseline testing except the (1) HS Omega-3 Index (OmegaQuant®) and meibum analysis. At 8 -weeks, the participants were reevaluated with all the testing conducted at the baseline and, in addition, a the Mastroda paddle was used, to collect meibomian gland secretions from each participant,

OUTCOME:

Based on OSDI which was taken at baseline, all twenty-one (21) participants reported a reduction of their primary complaint and fourteen (14) of the twenty-one (21) patients became completely asymptomatic

As illustrated in Table 1 , the participant levels of arachidonic acid, a direct precursor to pro-inflammatory eicosanoid derivatives, decreased significantly (p<.00004) from 12.2% at baseline to 10.3% at 8 weeks, as ^' measured in the blood. TABLE 1 RBC Hemoglobin - Oi»ega-6 (Arac do c Acid Decosapeataen ic Aek

Acid: C20:4n6~Arachidonic Acid; DPA ^::;: Docosapefitaenoic Acid; C22:5n6=Docosapentaenoic Acid)

The participant levels of EPA increased significantly (p<<00000) in the RBCs from baseline and at 8 weeks (0.8% and 3.2%, respectfully) and levels of DHA increased (p<.00349) in the RBCs from baseline and 8 weeks (3,3% and 4.1%, respecifuily), as shown in Table 2. Table 2 BC Hemoglobin - Omega-3 (DocosahexaenQic Acid/Eicosapentaeneic Acid)

(DH A ^:;;: Docosahexaenoic Acid; C22:6n3HDocosahexaenoie Acid; EPA-Eicosapentaenoie Acid; C20 :.5n3 ^:: -Eicosapentaenoic Acid)

Referring now to Table 3, the HS Omega~3 Inde Scores are provided for each of the twenty-one ( 1) participants.

Table 3 HS Omega-3 iad x Scores

(HS~Omega~3 index percentage - Red Blood Ceil Membrane Saturation of Omega-3s)

Tear osmolality decreased on average seventeen percent (17%) at the eight week exam period, as illustrated in Table 4.

Table 4 Tear Osmolality

(The osmolality of the right eye/left eye m miliiosmols)

As shown, there were variations in starting osmolalities among patients. The ase of topical drops within two (2) hour's of checking osmolalit disqualified participant 15 ^" s test as it may have had a dilution effect on the tears.

The lid margins were graded on a scale of trace - 4 for meibomian gland insipt sailors. The results of the participants of the clinical study are illustrated in Table 5,

Table 5 Lid Margins

„1 Q.

(Grading of meibomian gland appearance with reference to inSpissa&oii)

As shown, some patients did not have insipisation, but their lid margins were irregular versus smooth due to previous inflammation.

Referring now to Table 6, the improvement of Ί ear Break Up Time (TBUT) at eight weeks was statistically significant (p<,00027).

L a le

(Tear breakup time in seconds)

As shown, fifteen (15) of nineteen (19) participants demonstrated a lengthening of their TBUT from baseline.

As illustrated in Tables 7 and 8, meihuni analysis from, the initial samples from the study participants revealed that thirtee (13) participants had insufficient quantity of oil to analyze. Of the seve (7) thai were readable, none of the participants exhibited ornega-3 fatty acids in the roeibum. Bacterial components comprised 1.0 to 15 % of the oils present. Oleic acid (18:1 w9c) comprised betwee 34% and 60%.

Table 7 Meitam Analysis (BEFORE)

ECL deviates 0.000 !

and anti-iso represent bacterial coaiponents)

Ta le 8 MeibuiK A pfeihod P.LFA2

JSeqjName EH 412592 T [Start Time 4/12201115:18 s5artip# 5 jprof Method PLFA2 js m ID ll -SMIIH-04 ( i 08-CN) [Total Response 111929.9264 D# 4177 flotai ^" Named 71882.51325

6 percent Named 64.22099569 177 jTotal Amount 71928.91226 iCommeni

(DHA 22 :6w3, ,6j 12, 15 present at 3% post- treatment)

At the eight weeks exam period, fourteen (14) of the twenty-one (21) meibara samples had sufficient quantity to analyze. All fourteen (14) meibum samples had DHA (22:6n-3) present. The DHA was present as approximately 2% to 3% of the meibum composition.

Corneal staining was graded on a scale of trace, 1 +, 2+, 3+, and 4+. Improving one grade was considered clinically significant. Nine (9) of twenty-one (21) patients did not present with corneal staining at ^' baseline, but the eleven (11) patients that did all had significant improvement by way of slit lamp examination at the four week exam.

By end of the study, all participants showed improvement. Consequently, an increase in omega-3 RBC and meibum compositio had a direct correlation to the improvement of tear break up time, reduction in tear osmolality, and elevation of omega-3 index from the baseline. The stud also demonstrated the new presence of omega-3 fatty acids within the meibum itself.

An additional clinical study was conducted based on the following parameters:

OBJECTIVE:

To evaluate the clinical effect of the oral administration of a supplementation of omega-3 fatiy acids in re-ester ified triglyceride form on the meibum in patients suffering from symptoms of -dry eye and meibomian gland dysfunction.

SUBJECTS/VISITS/STUDY DESIGN:

Patients with meibomian, gland dysfunction were selected from the clinic and a meibum sample was obtained from each of the participants using a Mastroda paddle at baseline and at 8- weeks. The samples were immediately frozen and shipped at a later date on dry ice to be analyzed by the OmegaQuant® system. The participants were placed on a supplementation of omega-3 fatty acids comprising a daily dosage amount of 2,668 mg in a re-esterified triglyceride form (rTG) dispensed in four 667 mg capsules, each containing 420 mg of EPA, 140 mg ofDHA, 107 mg of other omega-3 's and 250 mg of Vitamin D(D3).

OUTCOME:

Of the eighteen (.18) available participant samples (three (3) samples appeared to be contaminated), twelve (12) showed an increase in the level of anti-inflammatory fatty acids (omega-3's) of almost five (5) fold and, more specifically, 4.85. The level of inflammatory fatty acids (omega-6 ^* s) decreased about two (2) fold. The results of the second stud confirm the findings of the first study showing an increase in. omega-3 in the meibum with the more accurate analytic system facilitated with by use of the OmegaQuant® system.

Furthermore, the findings of these studies indicate that on each blink a bath of inflammatory material, namely arachidonic acid (an omega-6) flows over the entire ocular surface. Here, lipases and other enzymes such as cyclooxygenase have the opportunity to break this chemical down into its prostaglandin derivatives, which are very potent inflammatory agents. When treated with oral supplementation of omega-3 in the re-esterified triglyceride form. the meibum is changed from an inflammatory bath with each blink to an anti-inflammatory bath. Reducing the inflammatory components about 2.5 fold, would have a profound effect on the tissues continually bathed by the meibum, changing to an almost five (5) fold increase in antiinflammatory would further stabilize the ocular surface. As taught by the present invention, bathing the ocular surface in an ami -inflammatory meibum instead of an inflammatory meibum is the mechanism of action of the supplementation of the omega-3 in re-esterified triglyceride form, delivered in the dosage amounts disclosed herein, such that said supplementation improved :">ά. the resolution of dry eye symptoms, tear osmolality, tear break up time, and blood saturation of omega.

The following examples will illustrate several embodiments of the present invention in further detail It will he readily understood that the nutritional or dietary supplement composition of the presen t in vention, as generally described and illustrated in the Examples herein, could be synthesized in a variety of formulations and dosage forms. Thus, the following more detailed descri ption of the embodiments of the methods, formulations and compositions of the present invention, as represented in the Examples are not intended to limit the scope of the invention, as claimed, but it is merely representative of various contemplated embodiments of the present invention.

EXAMPLE !

A daily dosage formulation of an embodiment of the nutritional or dietary supplement composition of the present invention administered for an increase in the ornega-3 level and a decrease in the omega-6 in the meibum composition of the meibomian glands is set forth as comprising:

Grnega-3 fatty acids

in certain embodiments of the present invention, a method for changing the quality of a meibum concentration of inflamed or dysfunctional meibomian glands comprises administering a supplementation comprising an effective amount of omega- 3 fatty acids as disclosed in Example I, wherein increasing levels of omega-3's and, respectively, decreasing levels of omega-6's in the meibum composition. Consequently, bathing the ocular surface in an anti-inflammatory meibuni instead of an inflammatory meibum is the mechanism of action of the supplementation of the omega-3's delivered in the dosage amounts disclosed herein so as to improve the resolution of dry eye .symptoms, tear osmolality, tear break up time, and blood saturatio of ornega-3. In certain embodiments, the omega-3 fatty acids comprise the esterified or re-esterified triglyceride form.

EXAMPLE

A daily dosage formulation of an embodiment of the nutritional or dietary supplement composition of the present invention administered for an increase in the ornega-3 level and a decrease in the omega-6 in the meibum composition of the meibomian glands is set forth as comprising:

Omega-3 fatty acids 1 ,000 mg - 3,000 mg

In certain embodiments of the present invention, a method for changing the q uality of a meibum concentration of inflamed or dysfunctional meibomian glands comprises administering a supplementation comprising an effective amount of omega-3 fatty acids as disclosed in Example H, wherein increasing levels of oinega-3 's and, respectively, decreasing levels ofomega-6's in the meibum composition. Consequently, bathing the ocular surface in a anti-inflammatory meibum. instead of an inflammatory meibum is the mechanism of action of the supplementation of the omega-3's delivered in the dosage amounts disclosed herein so as to improve the resolution of dry eye symptoms, tear osmolality, tea break up time, and blood saturation of omega-3. in certain embodiments, the omega-3 fatty acids comprise the esterified or re-esterified triglyceride form.

EXAMPLE 111

A daily dosage formulation of an embodiment of the nutritional or dietary ^' supplement composition of the present invention administered for an increase in the omega-3 level .and a decrease in the oniega-6 in the meibum composition of the meibomian glands is set forth as comprising:

omega-3 fatty acids 2,000 mg - 3,000 mg

In certain embodiments of the present invention, a method for changing the quality of a meibum concentration of inflamed or dysfunctional, meibomian gland ^' s comprises administering a supplementation comprising an effective amount of omega-3 fatty acids as disclosed in Example III, wherein increasing levels of omega-3 ^! s and, respectively, decreasing levels of om.ega-6's in the meibum composition. Consequently, bathing the ocular surface in an anti-inflammatory meibum instead of an inflammatory meibum is the mechani sm of action of the supplementatio of the omega-3 's deli ered m the dosage amounts disclosed herein so- as to improve the resolution of dry eye symptoms, tear osmolality, tear break up time, and blood saturation of omega-3. in certain embodiments, the omega-3 fatty acids comprise the esterified or re-esterified triglyceride form. EXAMPLE IV

A daily dosage fomrulation of an embodiment of the nutritional or dietary supplement composition of the present inventio administered for an increase i the omega-3 level and a decrease in the omega-6 in the meibam composition of the meibomian glands is set forth as comprising:

elcosapeniaenolc acid (EPA) > 600 mg

docosahexaenoic acid (DBA) > 500 mg

in certain embodiments of the present invention, a method for changing the quality of a meibu concentration of inflamed or dysfunctional meibomian glands comprises administering a supplementation comprising an effective amount of omega-3 fatty acids as disclosed in Example IV, wherein increasing levels of omega~3's and, respectively, decreasing levels of omega-6's in the meibimi composition. Consequently, bathing the ocular surface in an -anti-inflammatory meibum instead of an .inflammatory meibum is the mechanism of action of the supplementation of the omega-3 's delivered in the dosage amounts disclosed herein so as to improve the resoi ution of dry eye symptoms, tear osmolality., tear break up time, and blood saturation of omega-3. In certain embodiments, the omega-3 fatty acids comprise the esterified or re-esterified triglyceride form.

EXAMPLE V

A daily dosage formulation, of an embodiment, of the nutri tional or dietary supplement composition of the. present invention admimstered for an increase in the omega-3 level and a decrease in the omega-6 in the meibum composition of the meibomian glands is set forth as comprising:

docosahexaenoic acid (DHA) > 500 mg

In certain embodiments of die present invention, a method for changing the quality of a meibum concentration of inflamed or dysfunctional meibomian glands comprises administering a supplementation comprising an effective amount of omega-3 fatty acids as disclosed in Example V, wherein increasing levels of omega-3's and, respectively, decreasing levels of omega-6's in the meibum composition. Consequently, bathing the ocular surface in an anti "inflammatory meibum instead of an inflammatory meibum is the mechanism of action of the supplementation of the omega-3 's delivered in the dosage amounts disclosed .herein so as to impro e the resolution of dry eye symptoms, tear osmolality, tear- break up time, and blood saturation of omega-3-. In -77- certain embodiments, the omega-3 fatty acids comprise the esterified or re-esterified triglyceride form.

EXAMPLE VI

A daily dosage formulation of an embodiment of the ^■ ■.nutritional or dietary supplement composition of the present invention administered for an increase in the omega-3 level and a decrease in the omega-6 in. the meibum. composition of the meibomian glands is set forth, as comprising: eicosapeh ^' taenoie acid (EPA) > 600 nig

docosahexaenoic acid (DHA) > 500 mg

other omega-3 fatty acids 400 mg - 700 mg

in certain embodiments of the present invention, a method for changing the quality of a meibum concentration of inflamed or dysfunctional .meibomian glands comprises administering a supplementation comprising an effective amount ofomega-3 fatty acids as disclosed in Example VI, wherein increasing levels of omega-3's and, respectively, decreasing levels of omega-6's i the meibum composition. Consequently^ bathing the ocular- surface in an anti-inflammatory meibum instead of an inflammatory meibum is the mechanism of action of the supplementation of the omega-3 's delivered in the dosage amounts disclosed herein so as to improve die resolution of dry eye symptoms, tear osmolality, tear break up time, and blood saturation of omega-3. hi certain embodiments, the omega-3 fatty acids comprise the esterified or re-esterified triglyceride form.

EXAMPLE VII

A daily dosage formulation of an embodiment of the nutritional or dietary supplement composition of the present invention administered, for an increase in the omega-3 level and decrease in the omega-6 in the meibum composition of the meibomian glands is set forth as comprising:

eicosapentaenoic acid (EPA) 1,600 mg - 2,500 mg

docosahexaenoic acid (DHA) 500 mg - 900 mg

other omega-3 fatty acids 400 mg - 700 mg ί ^η certain embodiments of the present invention, a method for changing the quality of a meibuni concentration of inflamed or dysfunctional meibomian glands comprises administering a supplementation comprising an effective amount of omega- 3 fatty acids as disclosed in Example VII, wherein increasing levels of omega-3's and, respectively, decreasing levels ofomega-6's in the meibu composition. Consequently, bathing the ocular surface in an anti-infl mmatory meibum instead of an inflammatory meibum is the mechanism of actios of the supplementation of the omega-3's delivered in the dosage amounts disclosed herein so as to improve the resolution of dry eye symptoms, tear osmolality; tear break up time, and blood saturation of omega-3, in certain embodiments, the omega-3 fatty acids comprise the esterified or re-esterified triglyceride form.

EXAMPLE VIII

A daily dosage formulation of an embodiment of the nutritional or dietary supplemen composition of the present invention administered for an increase in the omega-3 level and a decrease in the omega~6 in the meibum composition of the meibomian glands is set forth as comprising:

eicosapentaenoic acid (EPA) > 600 mg

docosahexaenoic acid (DHA) > 500 mg

Vitamin D (as D3) 500 III - 2,000 IIJ

In certain embodiments of the present invention, a method for changing the quality of a meibum concentration of inflamed or dysfunctional meibomian glands comprises administering a supplementation comprising an effective amount of omega-3 fatty acids as disclosed in Example VIII, wherein increasing levels ofomega-3's and, respectively, decreasing levels of omega-6's in the meibum composition. Consequently, bathing the ocular surface in an anti-inflammatory meibum instead of an inflammatory meibum is the mechanism of action of the supplementation of the omega-3's delivered in the dosage amounts disclosed ^' herein so as to improve the resolution of dry eye symptoms, tear osmolality, tear break up time, and biood saturation of omega-3. In certain embodiments, the oinega-3 fatty acids comprise the esteri fied or re-esterified triglyceride form.' EXAMPLE IX

A daily dosage formulation of an embodiment of the nutritional or dietary supplement composition of the present invention administered for an increase in the omega-3 level and a decrease in the omega-6 in the meibum composition of the meibomian glands is set forth as comprising:

eicosapentaenoic acid (EPA) 1,600 mg - 2,500 mg

docosahexaenoic acid (DBA) 500 mg - 900 mg

Vitamin. D (as D3) 500 iU - 2,000 IU

in certain embodiments of the present invention, a method for changing the quality of a meiburn. concentration of inflamed or dysfunctional meibomian glands comprises administering a supplementation comprising an effective amount of omega-3 fatty acids as disclosed in Example IX, wherein increasing levels of omega-3 's and, respectively, decreasing levels of omega-6's in the meibum composition, Consequently, bathing the ocular surface in an anti-inflarnmatoiy meibum i nstead of an inflammatory meibum is the mechanism of action of the supplementation of the omega-3 's delivered in the dosage amounts disclosed herein so as to improve the resolution of dry eye symptoms, tear osmolality, tear break up time, and blood saturation of omega-3. In certain embodiments, the omega-3 fatty acids comprise the ester ified or re-estefified triglyceride form.

EXAMPLE X

A daily dosage formulation, of an embodiment of the nutritional or dietary supplement composition of the present invention administered for an increase in the omega-3 level and a decrease in the omega-6 in the meibum compositio of the meibomian, glands is set form as comprising:

eicosapentaenoic acid (EPA) > 600 mg

docosahexaenoic acid (DHA) > 500 mg

other omega-3 fatty acids 400 mg - 700 mg

Vitamin D (as D3) 500 IU - 2,000 IU

In certain embodiments of the present invention, a method for changing the quality of a meibum concentration of inflamed or dysfunctional meibomian glands comprises administering a suppiementation comprising an effective amount of omega-3 tatty acids as disclosed in Example X, wherein increasing levels of omega-3 ¾ and, respectively, decreasing levels of omega-6's in the meibum composition. Consequently, bathing the ocular surfa.ce in an anti¾flammatory meibum instead of an inflammatory meibimi is the mechanism, of action of the supplementation of the omega-3 's delivered in the dosage amounts disclosed herein so as to improve the resolution of dry eye symptoms, tear osmolality, tear break up time, and blood saturation of omega~3. In certain embodiments, the omega- 3 fatty acids comprise the esterified or re-esterified- triglyceride

EXAMPLE XI

A daily dosage formulation of an embodiment- of the nutritional ^' or -dietary supplement composition of the present invention administered for an increase in the omega-3 level and a decrease in the omega-6 in the meibum composition, of the meibomian glands is set forth as comprising:

eicosapentaenoic acid (EPA) 1 ,600 mg - 2,500 mg

docosahexaenoic acid (DHA) 500 mg - 900 mg

other omega-3 fatty acids 400 mg - 700 mg

Vitamin D (as D3) 500 IU - 2,000 IU

In certain embodiments of the present invention, a method, for changing the quality of a meibum concentration of inflamed o dysfunctional meibomian glands comprises administering a supplementation comprising an effective amount of omega-3 fatty acids as disclosed in Example XL wherein increasing levels of omega-3 's and, respectively, decreasing levels of omega-6's in the meibum composition. Consequently, bathing the ocular surface in an anti-inflammatory meibum instead of an inflammatory meibum is the mechanism of action of the supplementation of die omega-3 's delivered in the dosage amounts disclosed herein, so as to improve the resolution of dry eye symptoms, tear osmolality, tear break up time, and blood saturation, of omega-3. in certain embodiments, the omega-3 fatty acids comprise the esterified or re-esterified triglyceride

EXAMPLE XII

A daily dosage formulation of an embodiment of the nutritional or dietary supplement composition of the present invention administered for an increase in the omega-3 level, and a decrease in the omega-6 in the meibum composition of the meibomian glands is set forth as comprising: eicosapentaenoic acid (EPA) 1,650 mg - 1,750 mg doeosahexaenoie acid (DHA) 500 mg - 600 mg

other omega-3 fatty acids 400 mg - 500 mg

Vitamin D (as D3) 600 KJ - 800 10

in certain embodiments of the present invention, a method for changing the quality of a meibum concentration of inflamed or dysfunctional meibomian glands comprises administering a supplementation, comprising an effective amount of omega-3 fatty acids as disclosed in Example XIL wherein increasing levels of omega-3's and, respectively, decreasing levels ofomega-6's in the meibum composition. Consequently, bathing the ocular surface in an atiti-iiifiamraatory meibum instead of an inflammatory meibum is the mechanism of action of the supplementation of the omega-3 's delivered in the dosage amounts disclosed herein so as to improve the resolution of dry eye symptoms, tear osmolality, tear break, up time, and blood saturation of omega-3. In certain embodiments, the omega-3 fatty acids comprise the esierified or re-esterified triglyceride form.

EXAMPLE XIII

A dail dosage formulation of an embodiment of the nutritional or dietary supplement composition of the present invention administered for an increase in the omega-3 level and a decrease in the omega-6 in the meibum composition of the meibomian glands is set forth as comprising:

eieosapentaeiioic acid (EPA) 1,680 mg

doeosahexaenoie acid (DHA) 5 0 mg

other omega-3 fatty acids 428 mg

Vitamin (as D3) 334 I J

In certain embodiments of the present invention, a method for changing the quality of meibum concentration of inflamed or dysfonctiona! meibomia glands comprises administering a supplementation comprising an effective amount of omega-3 fatty acids as disclosed ^' in Example XIII, wherein increasing levels ofomega-3's and, respectively, decreasing levels ofomega-6's in the meibum composition. Consequently, bathing the ocular surface in an antiinflammatory meibum instead of an inflammatory meibum is the mechanism of action of the supplementation of the omega-3 ¾ delivered in the dosage amounts disclosed herein so as to improve the resolution of dry eye symptoms, tear osmolality, tear break up time, and blood saturation of omega-3. In certain embodiments, the omega-3 fatty acids comprise the esterifiedor re-esterified triglyceride form.

As concluded from the studies conducted, the supplementation of orhega-3's in. the re- esterified triglyceride form has a three (3) phase affect in the inflamed meibomian gland. First, the level of omega-3 is increased which, respectively, competes with arachidonic acid (omega-6) for binding sites on cyciooxygenase. Secondly, the amount of arachidonic acid which leads to prostaglandin synthesis is reduced. The products of COX 1 & 2 enzymes ^' working on omega-3 creates eicosanoids that compete with those in the prostaglandin pathway from the omega-6. Thirdly, the production of resolvin from the omega-3 may provide an even greater factor in the anti-inflammatory action within the meibomian glands. Consequently, the level of inflammatory fatty acids (omega-6's) decreased about two (2) fold and the level of anti-inflammatory fatty acids (om.ega~3's) increased: nearly five (5) fold.

As appreciated, surgery, by its very definition, involves the rapture of cell membranes. These cell membranes contain phospholipases- and omega-3 and omega-6 fatty acids. On a biochemical level, the pliospiiolipase liberates cyciooxygenase which is then available to act on the omega fatty acids. The omega-6 that is found in the cell membranes is known as arachidonic add and the irrflammatory pathway has been described hereinabove.

It follows that if the concentratio of omega-3, the anti-inflammatory fatty acid, as found in the compositions of the present invention, is at a higher cellular level then there will effectively be less inflammation, while taking into account there will correspondingly be less inflammatory substrate (arachidonic acid - omega-6's) since the area within the cell membrane in which these fatty acids reside is limited physically. Accordingly, the administration of compositions of the present inventionpreoperatively and/or postoperatively facilitate an increase in the anti-inflammatory level of omega-3 's in the cell membranes and correspondingly a decrease in the inflammatory level of arachidonic acid (omega~6's).

Although the composition and. methods of the present invention are directed, to any type of surgically invasive procedure where tissue damage occurs, one embodiment of the present invention, as described herein, contemplates the effects associated with cataract surgery. Moreover, the various Examples of the compositions that are outlined hereinabove are applicable, not only to an increase in the omega-3 level and a decrease in. the omega-ό level in the meibum composition of the meibomian glands, but also to reducing tissue after an invasive surgical proceeding. To this end, the foregoing Examples of the compositions of the present invention provide inherent advantages for reducing -inflammation in the tissue on a cellular level. A clinical study was conducted based on the following parameters:

OBJECTIVE:

To evaluate the clinical effect of the oral administration of a supplementation of oniega-3 fatty acids in the triglyceride form to patients having undergone an invasive: surgical procedure.

SUBJECTS:

in the preliminary study, five subjects or participants were included. Two of the participants had been taking about 2,600 tag of omega-3 fatty acids in the re-esterified triglyceride form daily prio to undergoing cataract surgery and three participants had not been taking any omega-3 supplements preoperati ely.

OUTCOME:

The clinical results of the study were, as follows, in the participants that were preoperatively taking about 2,600 mg of omega-3 fatty acids in the triglyceride form daily (and , more specifically, in the re-esterified triglyceride form), the level of aracMdonic acid (omega-6's) was reduced approximately about fifteen to twenty percent (15%~20%) lower than those participants who had not taken any omega-3 's (control group). Conversely, the omega-3 level was approximately twenty to twenty-five percent (20%~25%) higher in those participants that were supplementing with omega-3 fatty acids preoperatively.

It follows that an alternative treatment of postoperative inflammation of tissue having undergone a -invasive procedure by way ^' of administering a composition of thepresent invention preoperatively and/or postoperatively will reduce the amount of aracMdonic acid (omega-6 fatty acids) in the affected tissue. To date, those skilled in the art have focused efforts on blocking the cyclooxygenase enzyme so the arachidonic acid is not converted to prostaglandins and leukotrienes by means of applying a combination of NSAIDs and steroids. Whereas, NSAEDs and steroids are used to treat inflammation, they do so by blocking enzymes and, in particular, phospholipase for the steroids and cyciooxygenase by the NSAIDs, Conversely, the compositions and methods for using the same of the present invention have been found to reduce the amount of substrate for the reaction to achieve the same effects without the potentiall negative side effects associated with NSAIDs and steroids, by way of augmenting or replacing the substrate witli aft anti-inflammatory substance (omega-3 fatty acids) that further reduce the levels of arachidonic acid (omega-6's), thus tacliitating decrease in the levels of inflammation in the tissue. In this regard, the preoperative administration of the compositions of the present invention increase the anti-inflammatory levels of omega-3's in the cell, membrane thereby reducing the amount of arachidonic acid (omega-6's) affecting a reduction in tissue inflammation, whereby the use ofNSAIDs- and steroids may be eliminated or effectively reduced.

For example, one of the patients that was preoperative!}' taking omega-3' s had previously developed iritis, commonly referred to as inflammation of the eye. She was treated with topical steroids from which she developed caiaracts and elevated pressure. She was therefore refused cataract surgery because of the fear of postoperative inflammation. .As a participant in the clinical study, this, patient was evaluated b an. attending physician and permitted to undergo .cataract surgery in both eyes, while only using a minimal amount, of topical steroid (lotopredneiol)(lotemax). The topical steroids were delivered four (4) times a day initially, and then once a day the following week. As a result of the preoperative and postoperative supp lementation with the compositions of the present invention, thi s patient had only minimal postoperative inflammation.

Another example as to the effectiveness of the presen invention involved a. patient that had undergone photo refractive keratectomy (PRK). This patient experienced great difficulty healing the surface of her eye postoperatively, resulting in. significant haze in the cornea and poor vision in the 20/80 to 20/100 range. She had been receiving care from two (2) top ranked eye programs in the nation for a period of about eight (8) months with no measurable improvement. When referred to participate in a clinical study, she was placed on a composition of the present invention consisting of omega-3 fatty acids in the re-esterified triglyceride form. Within six weeks of supplementation, the patient's vision had improved to 20/30 to 20/40 range in both eyes. Moreover, her cornea clarity had. improved markedly. It was clear that omega-3 supplementation of the compositions of the present invention were having an effect on penetrating the cornea, which is an avascular tissue, to facilitate the healing outcomes.

Since the omega-3 is present in all cell membranes of the body, it follows that once a systemic level is achieved, generally felt to be over eight percent (8%) in the red blood cell index, this affect would occur relative to reducing tissue inflammation, associated with all types of surgeries, in fact one individual who had a blood level of ten percent 0%) was slated to undergo hip replacement surgery at a very large hospital. Th s individual was instructed to stop supplenienting with omega-3 's prior to surgery due to concerns about bleeding, Being well aware of the affects of omega 3, he refused to stop supplementing with omega-3 ⁵ s and proceeded to undergo the surgery. As a result, this patient's postoperative course was remarkable for lack of inflammation: and it was noted that he had recovered faster than anyone the clinic had ever seen.

As the clinical studies have shown, there is a correlation with a reduction in the level of OHiega-C¾ found in the cell membrane of tissue, when preoperative supplementation of composition of the present invention are administered. From a biochemical standpoint, the .supplementation of compositions of the present invention comprising an effective amount of omega-3 fatty acids function to displace or lower the levels of arachidonic acid (omega-6's), thereby facilitating a decrease or reduction in tissue inflammation after surgery. As taught herein, the administration of compositions of the present invention contemplate preoperativeiy and/or postoperatively.

i certain embodiments of the present invention, there was about a fifteen to twenty percent (15%-20%) decrease in the levels of arachidonic acid in tissue after surgery with the administration of the compositions of the present in ention preoperativeiy. Moreover, by administering the compositions preoperativeiy to participants in the study, it was shown that levels of omega-3 's increased about twenty-five percent (25%), whereby di^l d g the 15%~20% reduction in levels of omega-6's with this higher level of non-inflammatory omega-3's.

As contemplated herein, the composition and methods of the present invention comprise the steps of (!) administering preoperativeiy a composition consisting of a single fatty acid, wherein the single fatty acid consists of omega-3 fatty acids, (2) increasing levels of anti- mfiammatory omega-3 's in the tissue and (3) decreasing levels of inflammatory omega-6's (arachidonic acid) in the tissue, thereby reducing post surgical inflammation by reducing the prostaglandin precursors and increasing the anti-inflammatory and resolvins available at the surgical site. The methods of the present invention may furthe include the step of administering the composition of omega-3 fatty acids, as defined herein, postoperatively. Consequently, the compositions and methods of the present invention have been found to significantly reduce postoperative inflammation of tissue after surgery, thereby resulting in reduced post-surgical downtime and hospital stays and, most important, an improvement in healing outcomes. The present invention may be embodied in other specific forms without departing from its fundamental functions or essential characteristics. The described embodiments are to be considered in all respects only as illustrative, and not. restrictive. Ail changes which

come within the meaning and range of equivalency of the illustrative embodiments are to be embraced within their scope.