TR2021015389A2 - MICROEMULSION EYE FORMULATIONS WITH ANTI-INFLAMMATORY EFFECT FROM BLACK SEED OIL - Google Patents

Abstract

The invention relates to eye formulations containing black cumin oil and thymoquinone developed for use in the treatment of dry eye disease and their preparation method.

Description

DESCRIPTION MICROEMULSION EYE FORMULATIONS WITH ANTI-INFLAMMATORY EFFECT BASED ON BLACK SEED OIL Technical Field to which the Invention Concerns The invention relates to liquid eye formulations containing black cumin oil and thymoquinone developed for use in the treatment of dry eye disease and their processing method. State of the Art Related to the Invention (Prior Art) Eye; It is a small, complex multipart organ. Its anatomy, physiology and biochemistry are highly resistant to xenobiotics. The challenge for ASD was to deliver enough drugs to the area to treat ophthalmic diseases that bypass these protective barriers of the eye. The human eye consists of three layers that are suitable for transmitting and repellent Eg. The DE layer is called the hard layer or eye white. This layer bulges and forms the transparent layer E. The hard, white and stringy layer is a membrane that protects the eye. The vascular layer, which is a multi-vascular connective tissue, turns the eyeball into a complete dark room with its cell cover on both sides. In its anterior part, the ciliary region with the ciliary body muscles is located. Ciliary extensions are given to the extremities in the form of small pyramids that fill with blood to keep the multi-veined asphagium of the ciliary region tense. By extending the ciliary region, the vascular layer in the front part changes color and forms a diaphragm with a hole in the middle (pupil), this diaphragm is called the iris. The iris, whose color varies from person to person, contains muscle fibers that help enlarge and shrink the pupil. The retinal layer forms the third and very thin layer of the eye. The hollow, white small bubble in the back (visual nerve disc) is where the optic nerve enters and is called the blind spot. Just beyond your blind spot is SarEleke. This is the visual region where external images are best shaped. The optic nerve, which enters the posterior pole of the eye, spreads into many nerve fibers towards the vascular layer and ends with neurons arranged in three layers. The neurons in the first layer (multipolar neurons) continue in the cylinder axis, the optic nerve; In the anterior extension, the second layer of visual neurons abuts the cylinder axes. In this layer, there are cone- and rod-shaped neurons, one end of which enters the part of the network layer. Cones and rodsEi free endsAimed at the vascular layer. It affects the nerve endings of the cells coming to the vascular layer and the retinal cells. Obstacles in Ocular Drug Release The main obstacles and determining factors in ocular drug release are the physicochemical properties of the drug, excretion from lacrimal fluid, corneal barriers and extra-corneal absorption. Drug transport across the comeal epithelium occurs primarily through the paracellular or transcellular pathway. 11. Hydrophilic drugs penetrate primarily through the paracellular route, while lipophilic drugs prefer the transcellular route, passive or facilitated diffusion of light through the intercellular spaces. Lipophilic feature, solubility, molecular size and shape, charge and degree of ionization also affect the penetration route and rate in the comea. Most liquid dosage forms, such as solutions, suspensions, and liposomes, are either released from the conjunctival sac into the nasolacrine duct or cleared from the precorneal area, resulting in poor bioavailability of the drugs. Drugs are eliminated primarily from precorneal lacrimal fluid by solution drainage, lacrimation, and inefficient absorption into the conjunctiva of the eye. These factors and the corneal barrier prevent the penetration of the topically applied drug into the eye and prevent the applied drug from reaching the intraocular tissue. It is a transparent and curved tissue specialized to protect against external factors. The front surface of the cornea is the main visual component of the eye and the other one is the lens. The comea and the lens ensure that fluid coming from the external environment effectively reaches the retina. In humans, the cornea is an oval structure that makes up 1/6 of the external surface of the eye. When measured, the horizontal diameter is approximately 12.6 mm, the vertical diameter is approximately 11.7 mm, the thickness in the middle is 1.2 mm and the edge is 1.2 mm. The cornea had to be transparent for the Sgß to be clearly visible. Therefore, the structure does not contain blood vessels. The cornea creates a mechanical barrier that limits the path of drug molecules. It is thought that, due to its high lipid content, the epithelium and endothelium constitute an obstacle to the passage of hydrophilic molecules. Moreover, 5 liters between dead corneal epithelial cells. the existence of connections, It delays paracellular drug penetration, thus increasing the permeability of the cornea to hydrophilic and ionized molecules. On the other hand, the stroma, which consists of a cellular matrix composed of a layered arrangement of collagen fibrils, is characterized by a high water content, which makes this layer impermeable to lipophilic molecules. In general, to penetrate these three layers, the molecules appear to have an amphiphilic nature, characterized by the presence of distinct hydrophilic and lipophilic properties. The negatively charged cornea is more permeable to cations than anions at physiological pH. There is a transparent chamber in the chamber (anterior chamber) between the comea and the lens of the eye. This continuously produced transparent fluid is released into the blood through the Schlemm canal through the opening in the corner where the cornea and the iris meet. The pressure of this fluidity is regulated by our body. The inside of the eyeball is filled with a transparent substance (corpus vitreum) with a gelatin consistency. Comea; It consists of 5 layers: epithelium, Bowman's layer, stroma, Descement membrane and endothelium. Corneal epithelium: The cornea is embryologically of ectoderm origin, such as hair, nails and skin, therefore it is constantly renewed. This layer is the lowest layer of the cornea. It consists of five layers of cells. The cells at the bottom and the cells surrounding the cornea proliferate. These cells move towards the upper layers and replace the old cells, so the corneal epithelium is renewed approximately every two weeks. The comea epithelium is naturally lipophilic and acts as a selective membrane for small molecules, while also preventing the diffusion of macromolecules via the paracellular pathway. Lipophilic drugs can pass through the corneal epithelium transcellularly, and hydrophilic drugs can pass paracellularly. Outer outer layer (Bowman's membrane): It occurs as a result of irregular tightening of collagen fibers. Epithelial cells are tightly attached to Bowman's membrane and provide structural support to the comea. It is a protective layer against trauma and bacterial invasion. It is not a true membrane, and epithelial cells form Bowman's membrane and provide support to the comea. Corneal stroma Comea is a transparent structure that creates interest. It consists of approximately 100 layers of regularly aligned collagen fibers and has high hydrophilic properties. It contains sparse cells called keratocytes. When this layer is damaged, healing often leads to loss of transparency and/or changes in corneal curvature, negatively affecting vision. Ayn Elzamanda komean E transparent] [g E Elsags. Internal layer (Descement membrane): A basement membrane secreted by the corneal endothelial cells. 11. After damage or disease, the corneal endothelium is damaged and edema develops in the cornea. Corneal endothelium: The innermost layer of the cornea. It consists of hexagonal-shaped, single-layered cells. Cells are attached to each other with spheroids and function as a semi-permeable membrane. Thanks to the pump enzymes located on the lateral surfaces of the cells, the corneal stroma j water content is kept constant. These cells come into contact with intraocular fluid, not blood or lymphatic cells, and their embryological origin is different from the vascular endothelium. Tears l Tears; It is a layer with an ultra-thin aqueous-mucin gel structure, approximately 7710 μm thick, that covers the corneal and conjunctival epithelium, protects the eye surface, and contains electrolytes, water, mucin, vitamin A, and antimicrobial proteins such as lysozyme-lactoferrin. Its functions include eliminating small surface epithelial irregularities, turning the cornea into a smooth optical surface, moistening and protecting the sensitive surface epithelium of the cornea and conjunctiva, inhibiting the reproduction of microorganisms through mechanical washing and antimicrobial activity, and providing the cornea with the necessary nutrients. Tears are physical; Its average pH is 7.35 and varies between 5.20 and 8.35, its osmolarity is 302 mosm/L on average and its volume is 7 ul°. It is produced at an average of 1.2 uL per minute. Its refractive index is 1.136 "dE. Changes in blood glucose and urea are reflected in tears. 3. Under normal conditions, tears are isotonic. Mucin layer: It forms the innermost layer of the tear film layer. It covers the surface of the corneal and conjunctival epithelial cells on the surface. Kal EilEgEyaklasÜg is 0.5 microns. The mucin layer is secreted mostly by the goblet cells in the conjunctiva, conjunctiva and corneal epithelial cells, and in very small amounts by Henle crypts and Manz glands. The functions of the mucin layer are to protect it from trauma, - To turn the hydrophobic corneal epithelium into hydrophilic, thus allowing the tears to disperse on the ocular surface. - To reduce the surface tension with the help of the lipid layer and thus ensure the stability of the tear film. - To capture the shed epithelial cells, foreign substances and bacteria. Aqueous layer: It is located in the middle part of the tear film layer. The layer with the highest thickness is 11, approximately 8 microns thick. In the aqueous layer; It contains electrolytes, proteins, growth factors, vitamins, antibacterial molecules, cytokines, immunoglobulins and hormones. 1. This content provides nutrition and protection of the ocular surface. Functions of the aqueous layer: - To provide the appropriate electrolyte composition required for epithelial health, - To protect the ocular surface from infection, - To eliminate instantaneous irregularities on the comea surface, ° To wash the debris formed in tears, ° To regulate the functions of the comea and conjunctival epithelial cells, - To create an environment that enables cell movement. . Lipid layer: The most important function of the tear is to prevent the evaporation of the aqueous layer. It also prevents tears from spreading to the skin and is thought to have antibacterial properties. Lipid layer functions - Preventing evaporation of the aqueous layer - Smooth ocular surface - Contributing to clear vision by creating a hydrophobic barrier that prevents the tear barrier. OCULAR DRUG ADMINISTRATION WAYS Compared to drug administration to other parts of the body, drug application to the eye must overcome the significant difficulties presented by various ocular barriers. It is one of the most delicate ways of drug targeting due to the eye's unique physiology and physicochemical conditions. Common routes of drug administration to the eye are: topical, subconjunctival, intravitreal, retrobulbur, and intracameral. Topical application Topical application is most often applied to the eye chambers as ointments, gels, emulsions, or suspensions and is applied to the anterior segment. It is used in the treatment of diseases. Topical application has remained the most preferred method due to ease of application and low cost. Site of action for most topically applied drugs. Usually the cornea, conjunctiva, sclera and anterior segment, such as the iris and ciliary body, are the main obstacles to ocular drug targeting. Although delivering the drug to the retina via systemic administration would be ideal, it is still a challenge due to the blood-retinal barrier; The blood-retina barrier prevents the drug from passing from the blood to the retina. Oral application Whether used alone or topically, therapeutic drug concentration in the posterior segment could not be achieved with only oral drug use. Oral medication use, as opposed to the parenteral route, is the patient's preferred method in the treatment of chronic retinal diseases. However, for most of the targeted ocular tissues, high doses are required to provide accessibility and therapeutic efficacy at the desired dose, which makes oral administration unnecessary. These doses may cause systemic side effects and toxic effects. Vitamin A is an important nutrient found naturally in the tear film of healthy eyes. Vitamin A plays an important role in the production of the mucin layer, which is the innermost lubricating layer of the tear film, which is very important for the tear film. Vitamin A deficiency leads to mucin layer loss and goblet cell atrophy. Vitamin A drops protect the eyes from free radicals, toxins, allergens and inflammation. Topical retinoic acid therapy combined with systemic administration of Vitamin A has been used in the treatment of xerophthalmia. One or more single-acting retinoids can be dispersed in a pharmaceutically acceptable ophthalmic vehicle and applied topically for the effective treatment of dry eye disease. Today, oral supplementation with essential fatty acids is recommended by ophthalmologists. Essential fatty acids are precursors of eicosanoids, locally acting hormones that play a role in inflammatory processes. Essential fatty acids may benefit dry eye patients by reducing inflammation and changing the composition of meibomian lipids. Clinicians recommend the use of an omega-3 fatty acid diet to provide relief from dry eye disease. Examples of base lomega-3 gel capsules marketed specifically for dry eyes include Thera Tear and Bio Tears. Rasid et al. The Massachusetts Eye Research Institute has shown for the first time that topical application of a specific fatty acid is useful in treating the symptoms of dry eye disease. Topical α-linolenic acid treatment has been found to significantly reduce dry eye and inflammatory changes at both the ocular and molecular levels. Therefore, topical application of α-linolenic acid may be a new therapeutic method to treat clinical manifestations and inflammatory changes in dry eye disease. Pericucular and intravitreal application Although not the preferred method of patients, these methods are used to eliminate the inadequacy of topical and systemic application in the posterior segment. The periocular route includes subconjunctival, subtenons, retrobulbar and peribulbar administration and is less invasive compared to the intravitreal route. In subconjunctival injection, the drug is administered by passing the conjunctival epithelial barrier, which is a high-speed barrier for the permeability of water-soluble drugs. Drug solutions are applied close to the sclera and high retinal and vitreal concentrations are obtained. In contrast to periocular injections, intravitreal injection offers distinct advantages because the molecules are administered directly to the vitreous. Unlike other routes, intravitreal injection provides higher drug concentrations for the vitreous and retina. E. ACTIVE SUBSTANCE RELEASE, KINETICS AND BIOAVAILABILITY IN THE EYE Active substances enter the ocular tissues through the cornea or through the cornea. The corneal pathway plays an important role in diseases affecting the anterior segment such as inflammation, infection and glaucoma. The corneal epithelium acts as a fast-limited barrier. In the posterior segment, the blood-retina barrier prevents the active substance from leaving the blood and reaching tissues such as the retina. This barrier consists of the retinal pigment epithelium and retinal walls. Transport of active substance from tears to other tissues is initially affected by two factors; contact time of the transport system with the cornea: and the permeability of the cornea to the active substance. The active substance passes through the comea by passive diffusion, facilitated diffusion or active transport. Passive diffusion is not due to stones/EEIs. It is mostly affected by the physicochemical properties that determine the partition. Comea is a very flexible barrier and the permeability is 3310-7-10-5 om/sec7. The bioavailability of topically applied active substances cannot exceed 5%, even for small molecules. However, highly lipophilic substances (log P3) pass more difficult than substances with a log P (lipid-water partition coefficient 2-3). Because their passage through the lipophilic epithelium is limited by hydrophilic It slows down when it comes to the stroma. Comea permeability is the combination of passive diffusion and active transport. Active transport plays an important role especially in hydrophilic compounds. Eye studies are difficult to conduct in humans. That is why in Vivo studies are carried out in rabbits and pigs. As much as these models have human-like aspects, eye Various differences were noted, such as the frequency of blinking. In a study conducted with cyclosporin A, no difference was observed between humans and rabbits in terms of corneal permeability. It was determined that active substances applied to the eye had two basic pharmacokinetic models: one-compartment and two-compartment. In long-term determinations in the one-compartment model, one A number of deviations were observed. In this model, the active ingredient appears to penetrate the comea very easily. According to the accepted model that the eye consists of two basic compartments, the first compartment is the precorneal area and the second compartment is the aqueous humor. An active substance applied to the eye first encounters tears and then penetrates through the cornea. In a study conducted on rabbit eyes, the behavior of pilocarpine in the precorneal area was described as follows: - Precorneal area - Normal tear Evolution - Conjunctival absorption - Tear Evaporation D These factors affect the bioavailability of active substances in the eye and compete with absorption from the coma. The passage of the drug to the cornea is interrupted when the precomeal concentration falls below the concentration in the cornea. Drainage both controlled the dose and prevented the active substance from reaching peak concentration in the corneal epithelium. Structure Plan studies have shown that the first five minutes following the application of the active ingredient are very important. Bioavailability in eye preparations depends on the rate and degree of absorption of the active substance and is determined by four main factors; - Factors related to the application - Physiological factors - Factors related to the active substance - Factors related to the formulation Factors related to the application Although the application of hair at regular intervals increases the concentration of the active substance in the tear film, the advantage of being removed from the environment by lacrimal drainage is eliminated. When conventional eye preparations such as solutions, suspensions and ointments are applied to the eye, the eye tries to protect itself by activating its defense mechanisms. It has been stated that the human eye can hold 10 uL of 5 v under normal conditions. The dropper, which is generally used in eye drops, drops 50 to 75 uL of 5 v into the medium. However, the eye's ability to hold more heat than its tear volume is diminished. Thus, the concentration of active substance in tears decreases to an average of one quarter of the amount dropped into the eye, and a significant portion of the active substance is lost. was being overlooked. Additionally, if an ointment type preparation is applied, blurring of vision occurs. As a result, a bioavailability of around 1-10% is achieved from substances applied topically to the eye, which is a very low level and the patient has to apply the active substance frequently for effective treatment. This difficult situation also caused side and toxic effects to occur. Physiological Factors Human tears are renewed every 16 minutes. Various traumas such as inflammation, ulceration, and burns disrupt the integrity of the membrane. As a result, the eye becomes vulnerable to microorganisms and the passage of products from the damaged coma changes. Active substances bound to protein; Due to the size of the protein-active substance complex, it cannot pass through the corneal epithelium and thus its absorption does not occur. Some active substances may also undergo metabolism in the precorneal area. As in other biological substances, lysosomal enzymes are present in tears and can cause the breakdown of the active substance. Factors Related to the Active Substance Solubility and Partition Coefficient It is known that one of the most important factors affecting eye absorption is the solubility of the active substance in oil or water. For a substance that is highly soluble in oil, the stroma acts as a membrane, and for a substance that is highly soluble in water, the corneal epithelium acts as a rapidly silylating membrane. Therefore, lipophilic substances easily pass through the corneal epithelium, but resistance to hydrophilic substances is observed. Corneal epithelium is 50% resistant, stroma and endothelium are 25% resistant to lipophilic substances. Molecular weight and chemical form of the active ingredient: In general, it has been determined that compounds greater than 500 g/mol are less absorbed into the eye. It has been stated that those with a larger molecular weight do not have significant effects on bioavailability. Changing the chemical form, such as conversion to base, salt or ester form, affects solubility and therefore bioavailability. The prodrug approach is based on this principle. Conductors depending on the formulation: It is not possible to prepare eye preparations in a wide pH range. As we move away from physiological pH values, tear recognition increases and active substance loss occurs. The pH of all eye preparations is required to be close to neutral. If the pH shifts towards acid, tear secretion increases. At pH 7.4, many weakly basic active ingredients are non-ionized, therefore their absorption is high. Aids in the formulation provide better absorption by increasing the residence time in the eye or by causing chemical modifications or by reducing the surface tension of non-polar active substances. Some of these auxiliary substances also affect the integrity of the corneal epithelium. Desired properties in ocular formulations; Ensure bioavailability Prevent side effects Prevent side effects - Convenience to the patient and improve quality of life is the application method. Although topical drug application to the eye has many advantages, low bioavailability is considered a disadvantage in drug targeting due to many anatomical, physiological and biological factors that inhibit the penetration of drugs into the eye where they are available to protect the eye. Eye capacity can only remain in my hand. Excess SEJ, both normally produced and externally excreted, is rapidly discharged by the eye. In general, for patient compliance, it is thought that the preferred topical ocular drug delivery systems may be in the form of eye drops without causing additional vision or irritation. Conventional ocular drug delivery systems, such as eye drops, have very poor bioavailability because the eye is protected by a complex set of defense mechanisms that make it difficult to achieve an effective drug concentration within the target area of the eye. By increasing the viscosity of drug formulations applied to the eye, inefficient absorption can be eliminated. Increased viscosity may reduce drainage rates, prolong residence time in the precorneal area, and increase absorption in the eye. Drug delivery is open. Fleidan can be used to deliver pharmacological agents to four target regions of the eye - Vitreous cavity. Topical application to the eye can be used to deliver pharmacological agents to the anterior ocular, cornea and anterior / posterior regions. To treat disorders of the anterior structure of the eye, topical ocular application offers four main advantages over other routes of drug administration: ° Facilitates drug targeting into the eye, which is difficult to reach with systemic application. - It is a relatively simple and painless method to use. Topical drug applications to the eye are considered a noninvasive method due to absorption through the conjunctiva because the drug entering the conjunctiva reaches the general blood circulation rather than the intraocular segments. In addition to the conjunctiva, the nasolacrimal duct is known to contribute to the systemic absorption of the drug. Since systemic absorption does not constitute an obstacle to posterior segment targeting, the amount of intraocular drug obtained is generally below the effective concentration. DOSAGE FORMS APPLIED TO THE OCULAR SURFACE Solutions Ophthalmic solutions are sterile solutions applied to the eye by dripping. At the same time, other pharmaceutical factors such as the need for antimicrobial agents, osmolarity, buffering, Viscosity and appropriate packaging in these dosage forms are carefully examined. Polymer solutions increase the viscosity of the solution when polymer is added to the active substance solution, therefore they cannot pass through biological membranes, and since they can remain in the eye for a long time, they increase the bioavailability of the active substance by prolonging its contact time with the cornea. Different solution type systems can be prepared depending on the type and concentration of the polymer. These are mainly viscous solutions, bioadhesive hydrogels and in situ gelling systems. Emulsions Ocular emulsions are generally prepared by dissolving the active ingredient in an oil phase or by adding a dispersed surfactant and mixing with water to form an oil-in-water emulsion. The active ingredient can be added to the soluble phase at the beginning of the emulsion production process or after the emulsion has been prepared by a suitable dispersion process. can be added to the formulation. Suspensions Ophthalmic suspensions are used when the active substance is not dissolved in the desired solvent or its solution is not stable. However, suspensions have a disadvantage: Appropriate dosing may not be possible for the drug concentration to be dispersed in the solvent. The insoluble active ingredient in suspensions must be in a micronized form to avoid causing corneal irritation or scratching. Suspensions are commonly formulated by dispersing micronized drug powder (less than 10 µm in diameter) in a suitable aqueous solvent. Suspensions with particle sizes over 10 µm in diameter may result in a foreign feeling in the eye after application to the eye, causing reflex tear formation and damage to eye tissue. Reduction in particle size generally increased patient comfort and acceptability of suspension formulations. Ointments Ophthalmic ointments prolong drug contact time with the eye surface compared to many ophthalmic solutions, but ophthalmic ointments have disadvantages such as blurring of vision and dulling of the eyelids. Ophthalmic ointments can be partially sterilized or alternatively they must be produced from sterile ingredients in an aseptic environment. Use frequently filtration through a suitable membrane or dry sterilization. Ophthalmic gels consist of mucoadhesive polymers that enable local delivery of the active ingredient to the eye. These polymers can prolong drug contact time with biological tissues and thus improve ocular bioavailability. The choice of polymer plays a critical role in the release kinetics of the drug dosage form. Carboxymethylcellulose, carbopol, polycarbophil and sodium alginate can be given as examples. Inserts Ophthalmic inserts are coats of appropriate size and shape placed in the conjunctival fornix, lacrimal punctum, or cornea. Dosage forms EIE. This solid dosage form can be formulated as soluble (soluble) or as soluble (insoluble). Inserts ensure accurate delivery of the drug dose applied to the eye and can significantly increase its bioavailability in the eye. Contact lenses Traditionally used contact lenses provide a dose of medication for several hours. Conventional hydrogel soft contact lenses have the ability to absorb medication, minimizing its absorption through the lacrimal 5V or through the conjunctiva. Implants Implants are widely used to prolong the delivery of drugs to the ocular cells and tissues, especially in the posterior. INNOVATIVE DOSAGE FORMS APPLIED TO THE OCULAR SURFACE Hydrogels Hydrogels are systems in which polymers are used in their preparation. Synthetic polymers such as polyvinyl alcohol, polyvinyl pyrrolidone, polyethylene glycol (PEG), polyacrylic acid are used as substances with increased viscosity due to their physiological compatibility and physicochemical properties. As a result of the easy application of the formulation to the eye and its viscoelastic properties, it is ensured that it stays in the eye for a long time. These systems are called in situ gelling systems. When in situ gels are applied to the eye, a change occurs in the conformation of the polymers that are sensitive to external factors such as humidity, pH, ionic content and tear fluid in the eye environment, and as a result, the viscosity of the formulation changes. However, formulations that adhere to the mucosa have disadvantages such as blurred vision and an unpleasant sticking sensation, and patients' compliance with the treatment may be difficult. Colloidal Systems Colloidal system dosage forms include liposomes, nanoparticles, microemulsions, nanoemulsions, etc. There is. Colloidal dosage forms have advantages such as providing sustainable and controlled well-being in the targeted area, reducing the frequency of drug administration, and overcoming blood-ocular barriers. Encapsulating drugs with these colloidal compounds significantly increases the permeability in the targeted area and prevents degradation by ocular enzymes. It prevents. Nanosuspensions are suspended in a suitable dispersion medium stabilized by surfactants. They are submicron colloidal systems consisting of slightly water-soluble drugs. They are effective in increasing the solubility of the drug and thus increasing its bioavailability. The charge on the surface of the nanoparticle Elaril contained in the nanosuspensions facilitated adhesion to the comea. Nanoparticles are particles with a diameter of less than 1 µm containing various biodegradable/non-degradable polymers, lipids, phospholipids or metals. Liposomes are defined as small artificial vesicles that can be produced from natural non-toxic phospholipids and cholesterol. Also, dimensions: amphiphilic properties and Due to their biocompatibility properties, they are promising systems for ocular drug targeting. Niosomes are preferred over other vesicular systems for topical targeting because they are more chemically stable compared to liposomes. They can carry both lipotylic and hydrotylic drugs, and at the same time, they have low toxicity because they are nonionic. Dendrimers are macromolecular compounds consisting of a series of branches around the inner core. 11. Used in drug targeting due to their nanometer size ranges, ease of preparation and functionality, and the ability to display multiple copies of surface groups for biological diagnostics. 40`13years It was used by Hoar. Later J.H. In a study conducted in 1959, Schulman obtained a single-phase system by titrating a milky emulsion with hexanol and used the term "microemulsion" for this system. Microemulsions are optically isotropic and thermodynamically stable colloidal systems formed using oil, water and surfactant (YEM) and in some microemulsions, aerca yard Inc 1 surfactant (YYEM). There are three different types of microemulsions; water in oil (W/O), oil in water (Y/W) and continuous/transitional structures II. Microemulsions are systems that have small droplet size, are thermodynamically stable, transparent and increase the bioavailability of the active substance. It also overcomes toxicity problems by increasing the absorption and clinical effects of active substances with low solubility and bioavailability problems. Industrial production and sterilization are simple and cheap, and microemulsions, which provide convenience for soluble lipophilic drugs due to the lipophilic phase used, are the preferred drug delivery systems due to increasing the solubility of the active substance, ease of preparation and application, and long shelf lives. Microemulsions have low viscosity, the droplet size is larger than the micelle structure, and the radius of the internal phase droplets varies between 10 and 100 nm. After determining the substances to be used while preparing the microemulsion, the water phase and oil phase can be determined by visual observation and the comparison of the oil phase YEM and YYEM with the help of the triangular phase diagram. By adding YEM and YYEM together to the system, the interfacial tension is low: E. Microemulsions formed by using YYEM are called self-emulsifying microemulsions. Self-forming microemulsion systems consist of drug, oil and surfactant; They are biocompatible systems that usually contain one or more surfactants. Generally, in these systems, the lipophilic property of the active ingredient disperses between the aqueous and oily phases and the dispersed coefficient affects the pH of the solution. Self-micro/nano-emulsifying drug delivery systems are effective systems in improving drug solubility and absorption in drugs with solubility and bioavailability problems. Hydrophobic drugs can be dissolved in these systems. Microemulsions meet all possible requirements by being able to remain intact for a long time (thermodynamically stable), having low inter-surface tension and being able to form almost spontaneously, showing Newtonion flow properties due to their low viscosity, high solubilization capacity and small droplet size. . Microemulsions Ij Yap E 3 Organic phase: The choice of oil for microemulsions is important as it significantly affects microemulsion stability and is necessary to solubilize the large amount of lipophilic drug that needs to be administered. However, there is often no simple oil choice that meets both conditions perfectly. The organic phase not only dissolves the lipolyphic drug, it does the same! It also has various advantages such as increasing the absorption of the lipophilic drug from the targeted area. Typically original! Tar with long hydrocarbon chains prevents the formation of microemulsions. Because this chain structure prevents it from penetrating the interface film. Shorter chain oils can penetrate deeper into the hydrophobic tails of surfactants and thus form a more stable interfacial film necessary for microemulsion formation. However, the solubility capacity of the organic phase increases with the length of the chain. Therefore, choosing the most suitable oil to achieve microemulsion formation for a given surfactant and drug pair requires careful optimization. In addition to stability and dissolution issues, the ocular toxicity of oils also needs to be carefully evaluated during the formulation phase. However, in some cases, fat can act as the active component. It depends on the better solubility of the drugs! Instead of non-polar oils, polar oils such as medium or long chain triglycerides (MCT, LCT) are preferred. Additionally, organic phases should not be multipolar because they prevent microemulsion formation. MCT, which is more water soluble than LCT, is often preferred because it dissolves lipophilic drugs in high concentrations. Use at its purest: The outer phases include soybean oil, castor oil (glycerol trials, caprylics and caprylic acids), isopropyl myristate, fatty acids such as oleic acid, and sucrose esters such as mono-, di- or tri-palmitates of sucrose. Since these aids are well tolerated by the eye, their saline levels must be high to avoid contamination with potentially irritating substances. Aqueous phase: Water is the basic component in microemulsion formation. In O/W type microemulsions, it functions as a continuous phase and, contrary to its impermeability with oil, oil droplets are formed through hydrogen bonding with YEM and YYEM, and an oil-in-water type microemulsion is formed. Aqueous phase buffers are used to minimize the difficulties in preparing microemulsion systems and It should also be chosen to maximize microemulsion stability. Typically microemulsions should contain several additives such as antibacterial and isotonic agents. They can affect the available area of microemulsions and therefore microemulsions should be examined for the presence of other components. Salinity affects phase diagrams when ionic surfactants are added and non-ionic surfactants phase inversion reduce wetness (PIT) E. Play PIT if microemulsions are digested Ema conditions Eta fuel is very sensitive to humidity It is also important to set the initial pH at 7-8 to reduce the hydrolysis of phospholipids and triglycerides to fatty acids, which can reduce the pH of the microemulsion. Preservatives generally used in eye drop formulations cannot be included in microemulsions. It should not interact with surfactants and result in complexes and should not be absorbed into nanodroplets, which would significantly reduce bacterial activity in the bases. Thiomersal and chlorobutanol can be used in concentrations of 0.01-0.2%; Combinations: l Properties Surfactant (YEM): BEMs enable the formation of emulsions by reducing the tension between aqueous and oily phases. The preferred adsorption of YEM enables changes in the physicochemical properties of the interface due to its amnitic nature. While the YEM concentration is 0.1% by weight in emulsions, it is at most 10% in microemulsions due to the increase in the interfacial area between the aqueous and oily phase. This high concentration of YEM may cause ocular toxicity. Therefore, it may be better to reduce the amount of FEED and choose a digestion process that is not self-emulsifying. Generally, ionic BAITs are too toxic to be used for ocular application. Therefore, non-ionic BAITs should be preferred. These BAMS dissolve easily in water due to the presence of ether functional groups. The most commonly used BAMS for the digestion of microemulsions are poloxamers, polysorbates, polyethylene glycol and tyloxapol ( tylaxapol). Among PEG, PEG 200 is preferred due to its lowest viscosity. As for poloxamers, polyoxyethylene glycol blocks are hydrophilic while polyoxypropylene blocks are hydrophobic. The highest amount of oxyethylene poloxamer is found in 188" (82 w/w). In conclusion; It has the highest hydrophilic lipophilic balance (HLB) value in the poloxamer group (HLB: 29) and is preferred in the preparation of O/W emulsions. Finally, polysorbate 80 is the surfactant with both the highest HLB (HLBZIS) value and the lowest Viscosity. Amphoteric EMs such as lecithin are interesting due to their low toxicity. Before use, it is necessary to specify the degree of purification (change of fatty acid content) as well as the physical properties, composition and concentrations of phospholipids by decoction of this compound. Therefore, for example, lecithin obtained from egg yolk contains 32% phosphatidylcholine, 22% phosphatidylethanolamine and 19% phosphadylinositol. Miranol (MHT), another amphoteric FEED, is well tolerated in eye formulations. Auxiliary surfactant (YYEM): YYEM71s had three functions in microemulsion formation; (a) very low interfacial tension required for the formation of microemulsions and thermodynamic stability. (b) they can change the curvature of the interface depending on the relative importance of the nonpolar groups and (c) they affect the fluidity of the interfacial film. If the interfacial film is too hard, it prevents the formation of microemulsion and causes a more viscous double brittle phase. The addition of YYEM provides fluidity and branched It is equivalent to a YEM. It also enables the use of less concentration of FEED in formulations. The saturation of the YEMs on the hydrocarbon chain increases equally with the fluidity of the film. The YEMs used generally consist of small molecules such as alcohols with low molecular weight, glycols with carbon chain lengths varying between C2 and 310. Base alcohols such as pentanol and hexanol, lnl It cannot be used in pharmaceutical applications due to its irritating nature. Chained amines can also be used as YYEM. Microemulsions First Phase Studies 3 A widely accepted diagnosis for separating emulsions according to their equilibrium states was proposed by Winsor in 1947. Emulsions Winsor I, Winsor II, Winsor There are four types: III and Winsor IV. In the phase called Winsor 1, there are two phases, emulsion and oil. In this phase, there is excess oil in the upper part, and emulsion forms in the lower part. In Winsor II, there are 2 phases, water and emulsion. There is an emulsion in the upper part of SJ, and there is waste water in the lower part. 11. In Winsor II, there are 3 phases, these phases are W/O, Y/O and transitional emulsion phases. Here, the emulsion is formed in the middle part. In the Winsor IV type, there is only one phase obtained by homogeneously mixing oil, water and YEM. In emulsion systems, the concentrations at which the FEED/YEM-Oil-Water system gives the most suitable and durable 'Etch'bmulsion are determined by triangular phase diagrams. The triangular phase diagram is a method used to calculate the appropriate microemulsion area and the amount of the components in the formulation. It also helps investigate and prevent stability problems. When there are more than three components in the system, one or more of the components? It can be reduced to a variable number by keeping it at a constant rate. The diagram consists of the balance of W/W emulsion, lamellar or hexagonal silicrystals, SN microemulsion, O/W emulsion and O/W microemulsion. In Y/S and S/Y type microeinulsions, the droplet is surrounded by a film consisting of [clarÇ YEM and YYEM molecules. In the (W/Y) microemulsion, where the DLs' phase is oil, the hydrophobic part of the amphiphilic molecule is oriented towards the lis phase, and the hydrophilic lym high water droplet is oriented towards the g. These types of microemulsions are called reverse micelles. DG phase? In the water (O/W) microemulsion, the hydrophobic oil droplet of the amphiphilic molecule is directed into the oil droplet, and the hydrochloric acid is directed towards the outer phase. In transitional microemulsions, the ratio of water, oil and feed is equal. This type of microemulsion is multi-cornered and the oil and water phases are separated by a highly porous layer. Transitional microemulsions are formed as a monolayer by YEM9 in water and oil. The first step in the formulation of microemulsion drug delivery systems is the drawing of triangular phase diagrams. In the diagram drawn, the aqueous phase, oily phase, YEM and YYEM in the microemulsion formulation are properly mixed and the components in the formulation can be determined after the most suitable microemulsion area is determined. The most common and easiest method for drawing phase diagrams, which is closely related to the solubility of the components, is titration. One of the characteristic features of microemulsions prepared with non-ionic surfactants is that they are sensitive to change in shape. Each system is characterized by a narrow change in shape, and since it is known that the emulsion structure and properties of substances change with time, the PIT system has been developed, in addition to the HLB system. In this system, emulsifiers are graded according to the humidity at which they convert the O/W emulsion into a W/O emulsion. It is suggested in the literature that a diagnosis of HLB based on PIT is preferable. This siltivity is a feature of the FEED-oil-water phase diagram. For non-ionic YEMAs, the PIT is in a shelter and below this shelter, a YEM is preferably broken down as oil-swelling micelles into the water phase, and above this water glass, Lr is preferably broken down into water-swelling invert micelles into the oil phase. Hydrophilic-Lipophilic Balance (HLB) The hydrophilic-lipophilic balance system developed by Griffin in 1949 is used in FEED selection. The type of emulsion formed depends on the HLB value. HLB values of YEM and YYEMas vary between 0 and 20. As this value approaches 20, the emulsifier gains hydrophilic properties. Emulsifiers with HLB values less than 8 have lipophilic properties, while those with higher HLB values have hydrophilic properties. Microemulsion Preparation Methods In microemulsion formulation, aqueous phase, oily phase, YEM and YYEM are appropriately mixed; The resulting system can be O/W microemulsion, O/W microemulsion, O/W macroemulsion, O/W macroemulsion, lamellar or hexagonal crystal structure. One of the methods used in microemulsion preparation is high pressure homogenization. In the method, previously prepared macroemulsions are passed through a high-pressure homogenizer to obtain transparent microemulsions. In another method, PlT is used. In the method, W/W emulsion is prepared at 59-80°C with suitable components to prepare O/W microemulsion, phase separation occurs when the resulting emulsion is cooled to room temperature, and this phase separation is called sEaklEgE. Microemulsion can be obtained by adding YYEM to the system after phase separation. Microemulsions can also be created by using a high pressure homogenizer of systems prepared without YYEM. The most suitable amount of water for microemulsions can be determined by the titration method or the trial-and-error method. The trial-and-error method is not preferred due to the significant loss of substance and being an empirical method (77 In the selection of microemulsion components, care should be taken to ensure that the interaction of the components with each other is in line with the purpose. The most important issue in the preparation of microemulsions is that the substances that make up the system, especially the YEMs they contain in high amounts (20% on average), do not have toxic and irritating effects. Microemulsions 11 Characterization Optical Properties: Microemulsions are transparent because their droplet size is small and they pass the wavelength of visible LSLg. While transparency is achieved with a small amount of YEM in O/O systems, the amount of YEM required in O/O microemulsions is higher. Microemulsion stability is visible, no visible distortion, no physical appearance. It can be easily understood by observing the changes that occur. Depends on this As a matter of fact, optical examinations of microemulsions forming glassy structures have been carried out by direct imaging with scanning electron microscopy (TEM), albeit in black light. Droplet size determination: To measure the droplet size of microemulsions, different methods such as electron microscopy, Fifty-scatter EhJßANS and SAXS) and dynamic @lk hairEEnD (DLS) methods can be used. In wet scattering techniques such as SAXS and SANS, waves sent to the sample at a certain angle affect each other within the sample and spread the hair in all directions to create the pattern that will later create the appearance of the sample. Aerca structural properties are also revealed with the SANS method. Since the structural information is opened, the method, the diameters of the aggregates | It must also have the capacity to measure. For micelles and microemulsion droplets, neutrons and X-rays (wavelength <1.0 nm) can be used. The density measured in the narrow angle method affects the droplet structure and particle size data in proportion to the density of the substances causing the scatter, the formations between the droplets, and the structural differences between the droplets. In the DLS method, the sent Ether beam interacts with the colloidal particle [Ela Brown motion. The density of the correlation function gives the diffusion coefficient of the emulsion blood particles and hence the diameters of the microemulsion droplets according to the EiE Stokes-Einstein equation. Zeta potential analysis: Zeta potential analyzes are also performed in microemulsion systems. While N/S type emulsions with small droplets show low resistance, conductivity is observed when droplet coagulation is observed in W/Y type emulsions. Increasing resistance over time is a sign of aggregation, that is, durable. The system is diluted in bidistilled water with a conductivity of 50 pSem-l and the analyzes are evaluated according to the Helmholtz-Smoluchowski equation. In microemulsions used ocularly, internal phase droplets affect the loads. viscosity of the system It is of great importance to use single-point viscometers and multi-point viscometers in the measurement of viscosity. Rotation type rheometers using the cone-table method from multi-point viscometers were frequently used for functate & rheological analyses. While macroemulsions generally show pseudo-plastic flow, microemulsions show Newtonian flow. The microemulsion formulation requires certain rheological properties, especially in topical application. Ocular Use of Microemulsions: 1 Microemulsions have been developed as new drug delivery systems to increase the ocular absorption of ophthalmic drugs. Most formulation strategies aim to maximize ocular drug penneability by extending drug residence time in the corneal and conjunctival area while simultaneously minimizing precorneal drug loss. Microemulsions are promising dosage forms for eye use as they have many advantages regarding ophthalmic application. Microemulsions are of particular interest in delivering hydrophobic drugs to the cornea due to the possibility of loading the drug into the oil particle. O/W microemulsions have been evaluated as a suitable system for intraocular administration, dissolving poorly soluble drugs and increasing their absorption, as well as prolonging the release profile. Since the droplet size of microemulsions is very small (usually below 150 nm), these systems provide controlled release of the drug and provide greater penetration of the drug into the deeper layers of the eye. Although microemulsions have excellent advantages, the slender limas in the choice of YEM/YYEM and YEM/YEM The potential toxicity associated with higher concentrations often limits the usefulness of microemulsions. In recent years, the potential of microemulsions in ocular targeting has been examined by various research teams. Kumar et al. In 2014, they developed a voriconazole-loaded ocular microemulsion formulation and evaluated the formulation they developed and the traditional formulation in in vitro health and ex vivo migration studies. It has been observed that the microemulsion developed as a result of the study has superior properties than the traditional formulation. Fialho et al. In the study they conducted in 2004, they loaded dexamethasone into the microemulsion formulation. Stability, pharmacokinetics and ocular irritation test studies were carried out on dexamethasone loaded ocular microemulsion formulations and according to the results they obtained, the system they developed was superior to traditional systems. DRY EYE DISEASE Previously, dry eye was defined as a tear film disorder resulting from tear insufficiency or excessive tear evaporation, leading to initial symptoms of ocular surface damage and ocular discomfort in the interpalpebral space. Nowadays, this definition has changed and dry eye is defined as a complex inflammatory syndrome of the lacrimal functional unit. Dry eye syndrome, for which the terms "dysfunctional tear syndrome" or "lacrimal keratoconjunctivitis" may also be used, refers to the presence of clinical or subclinical inflammation on the ocular surface. In 2007, a new definition for dry eye, including a comprehensive classification system, was reported by the International Dry Eye Study Committee. A three-part classification system was published in the committee. The first part addresses etiopathogenesis and identifies the multiple causes of dry eye. In this group, dry eye is divided into two main categories; aqueous insufficiency (Sjögren's or non-Sjögren's related) and evaporative disorders. The second part is mechanistic and shows that each cause acts along a common pathway (tear hyperosmolarity and tear film instability). The third part is based on the severity of the disease (visual disorders: conjunctival injection, conjunctival staining, corneal staining, eyelid findings, tear time, Schinner test, and provides a rational basis for treatment (90). The mechanisms in dry eye formation are tear hyperosmolarity, tear transport instability and inflammation. Tear hyperosmolarity may occur due to increased evaporation or decreased aqueous secretion. The increase in protein and electrolyte concentration causes a decrease in tear volume, which initially leads to irritation of the ocular surface and inflammation in later stages. Tear film instability, diagnosed by a decrease in eye dryness over time, is also increased by evaporation. it is related to sma Dry eye is a condition frequently encountered in ophthalmology clinics. According to recent studies, the prevalence of dry eye in various age groups is estimated to be between 5-30%. There are 7.1 million people over the age of 40 in the United States. While it is stated that jocular discomfort due to a person's dry eye syndrome is prohibited, other studies on prevalence show that the population over the age of 40 increases with age and is significantly more common, especially in women. The incidence of dry eye in various systemic diseases increases significantly. Etiology of Dry Eye Different and interrelated factors may play a role in the etiology. Decrease in the amount of tear production may develop due to disorder in the distribution of tear fluid on the ocular surface, irregularity in the corneal epithelium, and pathologies in the tear lipids. Aqueous tear insufficiency occurs due to the inability of the tear gland to secrete enough tears. As tear quantity decreases, osmolarity increases, and hyperosmolarity triggers inflammation, increasing the severity of dry eye. In Sjögren's syndrome, there is an autoimmune cause that causes damage to the glandular structure as a result of lymphocyte infiltration in the tear and salivary glands. As a result, tear and saliva secretion decreases. Aqueous insufficiency not related to Sjögren's syndrome may occur due to reasons such as aqueous insufficiency caused by aging, congenital absence of the lacrimal gland (Congenital Alacrima), impairments in the sympathetic and parasympathetic innervation of the lacrimal gland, infiltration and ablation of the lacrimal gland. In dry eye due to excess evaporation, tear flow from the tear gland is normal, but dry eye clinic develops as a result of evaporation from the ocular surface. Reasons such as meibomian gland dysfunction (posterior blepharitis), prolonged interval between blinking, low humidity in the environment, allergic conjunctivitis, contact lens use and drugs toxic to the ocular surface cause dry eye, especially by deterioration in the lipid layer and changes in lipid components. Dry Eye Mechanism The basic mechanisms of dry eye are driven by tear hyperosmolarity and tear film instability. Tear hyperosmolarity activates a series of inflammatory events on the ocular surface and damages the surface epithelium through the contact of inflammatory mediators with tears. Epithelial damage includes cell death by apoptosis, loss of goblet cells, and disruption of mucin expression, leading to tear film instability. This imbalance exacerbates ocular surface hyperosmolarity and cEE completes the cycle. Tear film instability can be initiated by a variety of etiologies, including xerophthalmia before tear hyperosmolarity, ocular allergy, use of topical preservatives and contact lens wear. Epithelial damage caused by dry eye: corneal nerve endings are sensitive, leading to compensatory reflex lacrimal tear secretion due to increased eye opening and closing. Deficiency of normal mucin on the ocular surface also contributes to symptoms by increasing the frictional resistance between the eyelids and the eyeball. The high reflex input during this period has been suggested as the basis of a neurogenic inflammation within the gland. The main causes of tear hyperosmolarity are watery tears resulting from lacrimal insufficiency and/or increased evaporation from the tear film. This condition is indicated by the arrow in the upper middle part of the figure. Recording as a result of increased evaporation is favored by environmental conditions of low humidity and high air flow and may clinically result from muibomian gland dysfunction leading to a particularly unstable tear film lipid layer. Lid lipid quality may change with the effect of esterase and lipase enzymes released in normal lid products, with increasing numbers of blepharitis (eyelid inflammation and stye). Decreased watery tears were due to impaired conduction of lacrimal flow into the conjunctival sac. It is unclear whether this is a feature of normal aging. However, it can be induced by some systemic drugs such as antihistamines and antimuscarinic agents. The 11 most common causes are inflammatory lacrimal damage, which is seen in autoimmune disorders such as Sjögren's syndrome, as well as in dry eye without Sjögren's syndrome. Inflammation causes both tissue destruction and a potentially reversible neurosecretion. A receptor blockade may be caused by circulating antibodies to the M3 receptor. Inflammation is favored by low tissue androgen levels. Tear conduction may be inhibited by cicatricial conjunctival scratching or reduced by loss of sensory reflex drive from the ocular surface to the lycrimal gland. Eventually, chronic surface damage of dry eye leads to a decrease in corneal sensitivity and a decrease in reflex tear secretion. Various etiologies may cause dry eye syndrome, at least in part by the mechanism of refractive secretion blockade: refractive surgery, contact lens wear, and chronic abuse of topical anesthetics. Treatment of Dry Eye The first things to do in the treatment of dry eye are; The aim is to identify and treat the underlying causes, to eliminate the symptoms, to reduce tear osmolarity, to improve tear layer stability and to reverse eye surface damage. Regardless of the cause of dry eye syndrome, the patient must be informed about the treatment. Treatment should aim to improve symptoms, ensure the integrity of the ocular surface, and prevent complications. If there is a systemic disease that causes dry eye, it should be treated. It should always be kept in mind that dry eye is a chronic, often resistant disease that can be very difficult to treat definitively. Since dry eye symptoms are found in a wide range and are common in many ophthalmological diseases, mistakes can be made in the treatment of dry eye. Since there have been studies in recent years indicating that this disease is an inflammatory syndrome, treatment protocols have begun to shift to anti-inflammatory and immune modulator areas. TO. Medicines used in the treatment of dry eye disease are divided into two. These; - Ophthalmic Lubricants Ophthalmic anti-inflammatory agents: Ophthalmic anti-inflammatory agents are applied to the eyes to reduce pain or inflammation. These are products formulated to treat. Anti-inflammatory agents act against one or more mediators that cause inflammation and reduce irritation and swelling in the eyes. Also use anti-inflammatory eye drops to relieve pain after surgery, Jim. Restasis® Ophthalmic Emulsion Restasis ophthalmic emulsion contains 0.05% Cyclosporine as the active ingredient and castor oil glycerin, polysorbate 80, carbomer, water and sodium hydroxide as auxiliary ingredients. Restasis eye drops contain Cyclosporine, an immunosuppressive drug. Cyclosporine may increase tear production, which reduces inflammation in the eyes. Restasis eye drops are used to treat chronic dry eye that may be caused by inflammation. Xiidra® Ophthalmic Solution Xiidra ophthalmic solution contains 5% fibertegrast as the active ingredient and sodium chlorite, sodium phosphate, sodium thiosulfate, sodium hydroxide, hydrochloric acid and water as auxiliary ingredients. Xiidra works by blocking a certain protein on the surface of cells in our body. This protein can cause your eyes not to produce enough tears or to produce tears that are not the right amount to keep your eyes healthy. Xiidra eye drops use to cure dry eye disease g el symptoms i Elli. Ophthalmic lubricants: Ophthalmic lubricants are used to treat dry and irritated eyes. 1H1t Black Seed Our country has a very wide range in terms of agriculture and plant diversity. Among this plant variety, black cumin is a Nigella sativa (NS) species found in the Ranunculacea family. It is also known as black seed, black cumin, and grain of abundance. 12 Nigella species grow in our country, and among these, the seeds of Nigella sativa L., Nigella damascena and Nigella arvensis are used as food and medicine. Nigella sativa L. is the most widely traded species in our country. Black cumin is a cultural plant that has been used medicinally since ancient civilizations. Especially the seeds of black cumin, which is used both as food and as a folk medicine, have recently been used as a traditional medicine in the Middle and Far East for a long time. It has been traditionally used in the treatment of asthma, bronchitis, inflammation, eczema, fever, flu, hypertension, cough, headache, dizziness, diabetes, kidney and liver dysfunction, nerve diseases, rheumatism, cancer and gastrointestinal diseases. In studies investigating the pharmacological effects of the components contained in black cumin seed, it has been reported that they have anti-inflammatory, antimicrobial, antibacterial, anticarcinogenic, antitimoral, antioxidant and immune system strengthening effects. Black cumin seed generally contains fixed oils, volatile oils, protein, amino acids, carbohydrates, fibers, alkaloids, tannins, saponins, minerals, ascorbic acid, thiamine, niacin, pyridoxine and folic acid. Fixed oil contains saturated fatty acids myristic acid, palmitic acid and stearic acid, and unsaturated fatty acids oleic acid, linoleic acid, eicosadienoic, arachidonic acid and linolenic acid. The essential oil, which contains the most important active components from a pharmacological point of view, includes Thymoquinone (TQ), Dithymoquinone (DTQ), Thymohydroquinone (THQ) and Thymol (TMYY, as well as nigellon, carvacrol, p-cymene, d-limonene, (1 and [3- It contains pinene. CHEMICAL PROPERTIES OF BLACK SEED Black cumin seed contains 34-39% oil and more than 307% of it constitutes fixed oil and fraction. In addition to the essential active components of essential oil, carvacrol, nigellimine-N-oxide, nigellisine and nigellidin. Black cumin seed also contains eight of the nine essential amino acids. It has been reported that there are four basic active pharmacological substances in the essential oil of black cumin seed, namely TQ, DTQ, THQ or nigellon and THY. The most important of these is the continuous studies on them recently. yapHdEgETQ has the structure 2-isopropyl-5-methyl-1,4-benzoquinone and its molecular formula is C10H1202. Various pharmacologically active chemical compounds have been isolated from the essential oils of black cumin seed; mainly TQ, DTQ, THQ and TMY YanDsEa; p-cymene, d-limonene, a-pinene, ß-pinene, trans-anethole, carvacrol and nigellone. TQ, p-cymene, longifolin, carvacrol, u-cubebene, (i-pinene, limonene, ß-pinene and sabinene are the main components of black cumin seed essential oil. Linoleic acid, oleic acid, palmitic acid are the main fatty acids in fixed oil. Black cumin seed. The seed has a very high content of saturated fatty acids, and the majority of the unsaturated fatty acid is linolenic acid, along with oleic acid, which is less partial. It contains saturated fatty acids such as palmitic, stearic, eicosadienoic and myristic acid. Black cumin seed has a rich content of vitamins and minerals. Vitamins Bl (thiamine), B2 (riboflavin), B3 (niacin), B6 (pyridoxine), B9 (folic acid), OL and y-tocopherol, retinol, as well as potassium, iron, bakE, calcium, zinc, phosphorus, magnesium and There are also necessary mineral substances such as selenium. E. PHYSICAL CHEMICAL PROPERTIES OF BLACK SEED OIL In many studies, the physicochemical properties of black cumin seed oil have been examined with parameters such as fatty acid content, color, viscosity, thermal property and oxidative stability. It has been argued that UV absorption is high due to its color composition and therefore it can provide a sun protective effect. Since it contains high amounts of natural antioxidants and low amounts of polyunsaturated fatty acids, it has a distinctive lead over different vegetable oils. It has been reported that the total phenolic component content of black cumin oil obtained by the cold press method is high and low oxidative stability is observed. THIMOCINONE TQ, which is found at a rate of 18-24% in the essential oil of black cumin seed and has a molecular weight of 164.2 g/mol, is a main active phenolic compound and is widely used in the treatment of many patients with some pharmacological effects. In vitro and In vivo studies have shown that it has versatile biological activity properties and its toxicological effects have been examined extensively. Enol and keto forms exist in ltautomeric form, and the keto form is responsible for its pharmacological properties. TQ is a hydrophobic molecule, so it enters the cell by passive diffusion. It has high sensitivity against lsfga. Its stability decreases at high pH and minimal degradation occurs at acidic pH. ANTI-INFLAMMATORY PROPERTIES OF THIMOKINONE Many effects have been observed when the pharmacological properties of TQ are investigated. TQ is used in treatment areas such as anti-inflammatory, antimicrobial, antibacterial, anticancer, nervous system, respiratory system, antidiabetic and antioxidant. The properties of these main components with pharmacological properties, such as TQ, THQ, DTQ, TMY, as well as p-cymene, nigellidine and nigellisin contained in black cumin oil, are stated in the literature. Inflammation, cyclooxygenase (COX) and lipoxygenase (LUX). It is mainly regulated by two enzymes. While prostaglandins (PG) are synthesized in the COX pathway, locotrienes (LT) are synthesized in the LOX pathway. TQ inhibits both COX and LOX pathways. This reduces the formation of reactive oxygen species (ROS). The anti-inflammatory effect of black cumin seed has many benefits. Acetic acid and formalin tests were applied, oil analysis was done by GC-MS (gas chromatography 7 mass spectroscopy) method, 20 different compounds were identified in the oil and the anti-inflammatory effects of the obtained essential oil were determined as 37.3% and 13%, respectively. P-cymene and TQ were found to be the compounds responsible for the effect with ratios of 7. Houghton et al. reported that black cumin seed fixed oil and TQ, as a potential anti-inflammatory substance, inhibit the inflammation chain at some stages, inhibit eicosanoid structure and membrane lipid peroxidation. It is shown with a play he made. It has been observed that black cumin seed fixed oil and TQin inhibit calcium ionophore-stimulated arachidonate metabolism, COX and 5-LOX pathways in the white blood cells of stimulated mice. These results have also shown that the anti-inflammatory effect may be due not only to the TQ in the oil but also to other components, especially the 3202 fatty acid. In this study, the anti-inflammatory effect of the plant seeds EiEi aqueous extract was demonstrated with its inhibitory properties on paw edema in animal models. Edema was created in mice with carrageenan, and it was observed that black cumin extract given orally partially eliminated the edema, and intraperitoneal administration completely eliminated the edema. In another study, edema was created in the paws with carrageenan and in the ears with castor oil, and black cumin essential oil and its effectiveness were observed in mice. It has been determined that the component TQ7 has a dose-dependent anti-inflammatory effect and prevents the formation of edema and granulomas. The anti-inflammatory activities of DTQ, THQ, TMY and TQ compounds were investigated in vitro using COX-1 and COX-2. The results showed that all products were against at least one form of COX. It showed that it had more significant inhibitory activity compared to indomethacin. This shows that DTQ, THQ, TMY and TQ collectively contribute to the anti-inflammatory activity of black cumin and that a new non-steroidal anti-inflammatory drug can be developed by further studies. Some synthetic drugs used for the treatment of inflammation cause changes in biological activities and have adverse effects. In the studies conducted to date, TQ It has been proven that black seed has a cytoprotective effect and this effect is mainly associated with the anti-inflammatory effect of TQ. Components of black cumin have been shown to have an anti-inflammatory effect in many inflammatory diseases, including experimental allergic encephalomyelitis (EAE), colitis and arthritis. EAE is higher when the animal receives DTQ. They showed that glutathione levels and perivascular inflammation showed no disease symptoms compared to animals not treated with EAE. These data demonstrate the therapeutic potential of TQ in the EAE model and suggest that it may have possible effects in the treatment of multiple sclerosis in humans. Kapan et al. They investigated the effectiveness of TQ against bacterial translocation and inflammation stimulated by mechanical intestinal obstruction. In the experimental study, in the group given TQ in rats, intestinal inflammation decreased the secretion of inflammatory cytokines, oxidative damage and bacterial translocation in the intestine and liver. It was concluded that it actually prevents inflammatory changes and is beneficial in remodeling the mucosa. they did. They reported that TQ successfully controls bacterial translocation and is effective in improving intestinal barrier function. In a clinical study, patients with rheumatoid arthritis were treated with black cumin seed oil at a dose of 2 g/day for 2 months, and erythrocyte sedimentation, plasma bake, uric acid, creatinine, alanine aminotransferase (AST) and asparate aminotransferase (ALT) levels were improved. A decrease was observed, an increase was observed in plasma zinc, vitamin E and vitamin C levels, and anti-inflammatory activity was detected. TOXICITY STUDY OF TIMOKIN0N9 Darakhshan et al. In the study on TQ, the therapeutic effects of TQ were observed. It was reported that TQ could correct oxidative stress-induced damage in many organ systems such as kidney, liver, heart, lung and stomach. Many beneficial effects of TQ, such as anticarcinogenic, anti-inflammatory, analgesic, hypoglycemic, and immune system strengthening, have been suggested and it has been reported to be safe. The toxic effects of TQ have only been demonstrated at very high doses. TQ has a selective cytotoxic effect at high doses. It has been reported that TQ can effectively kill tumor cells without showing cytotoxicity in normal cells such as osteoblasts, mouse kidney cells, human lung fibroblasts and vero cells. It has been reported that TQ does not show toxic and lethal effects in rats when taken orally at doses of 10-100 mg/kg. In the same study, the maximum tolerable dose of TQ was found to be 22.5 mg/kg in male rats and 15 mg/kg in female rats in IP injection, and 250 mg/kg in male and female rats orally. In the same study, in acute toxicity tests, the oral lethal dose (LD50) value of TQ in mice (1.52-3.77 operating range) was reported as 2.4 g/kg and the IP LD50 value in rats was reported as 10 mg/kg. However, Mansour et al. In another study conducted by et al., it was reported that TQ 4, 8 did not change biochemical parameters and showed a toxic effect in IP injection at doses above 50 mg/kg, and the IP LD50 value was 90.3 mg/kg. In the study conducted by Khader et al. If TQ3 does not undergo biotransformation in the body, ROS formed in it can increase oxidative stress. High doses of TQ can consume antioxidant enzymes and cause DNA damage in hepatocytes. B.H. Ali et al. reported that black cumin seed and oil have low toxicity in their study on black cumin seed and oil. TQ is the route of administration. They said that it affects the results of toxicity. It has been reported that high doses of TQ cause liver damage in the In Vivo study. Acute administration of high doses of TQ (2 g/kg and more) caused hypoactivity and respiratory difficulty. In Abukhader studies, IP administration of TQ should be achieved through systemic circulation. It has been shown to cause acute pancreatitis as a result of IHA. Male and female levels of TQ were monitored for 5 days following a single dose IP injection of 20, 30 and 40 mg/kg. Dosing of 30 and 40 mg/kg of TQ Eonras] Irritability within 72 hours showed signs of toxicity such as insomnia, piloerection, and mild abdominal swelling. An average decrease of 17.3 g in body weight was observed in rats on the 3rd and 4th days of the experiment. It was reported that no symptoms were observed in female rats administered 20 mg/kg TQ. TQ was observed in males and females. Levels were monitored for 5 days following a single oral dose of 200, 300, and 500 mg/kg. Within forty-eight hours, an increase in weight, diarrhea, hypoactivity, mild abdominal distention, and shortness of breath were observed. Subsequently, the weights of the lipids returned to normal and signs of toxicity continued on the 5th day of the experiment. has disappeared until now. Death occurred in one male and one female rabbit at the 500 mg/kg dose. In the study conducted by Ezzat S. El Daly, it was shown that black cumin seed extract (50 mg/kg) did not affect kidney and liver function biomarkers when administered IP to mice for 5 days. A study by Zaoui et al. [Emada, fixed oil of black cumin] at a dose of 10 mL/kg It has been reported that oral administration did not cause mortality or histopathological changes or significant changes in basic liver enzymes including ALT, AST and glutathione (GSH) in rats for up to 12 weeks. In the same study, it was done by giving 2 mL/kg/day oral dose of black cumin oil to the rats for 12 weeks. In a chronic toxicity test, no histopathological modifications or changes in liver enzyme levels including ALT, AST and GSH were observed. Serum cholesterol, triglyceride and glucose levels and platelet and leukocyte amounts were found to be significantly lower compared to control values. Hemoglobin and hematocrit values were found to be significantly higher. Additionally, A decrease in body weight was observed in the group given black cumin oil compared to the animals in the control group. Gali-Muhtasib et al. According to the study, TQ did not cause death or change in average body weight in Balb/c mice when administered for 20 consecutive days, and it was reported that TQ was well tolerated in mice. In a study by Shoji Fukushima et al., 200 mg/kg/day of muffin for 14 weeks. It was reported by Al-Ali et al. that male selans given with black cumin oil did not cause pathological changes in the liver, kidney, spleen and other organs, and that black cumin did not affect blood biochemistry in mice. The IP LD50 value of TQ in mice was 104.7 mg/kg and the oral LD50 mg/kg. It is reported as . Zaki Tubesha et al. did g. In the study, the acute toxicity of the nanoemulsion formulation of TQ was investigated in male and female silanes. The preparation containing 44.5 mg TQ was administered quickly orally. Subsequently, the animals were examined for 14 days. All animals were recorded as healthy. Mortality and any toxicity were not observed. General behavior, body weight No change was determined in food and water consumption, organ network hematology, histopathology and clinical biochemical parameters. It was concluded that the nanoemulsion formulation of TQ containing 44.5 mg TQ was not toxic. In the study conducted by E. Islam SN et al., it is reported that TQ is a contact allergen. In rats exposed to 2.5 uL black cumin oil, twice a week for 4 weeks; It has been reported that it reduces neutrophils and splenocytes and increases peripheral lymphocytes and monocytes. Khader et al reported 2 cases of allergic contact dermatitis with maculopapular eczema symptoms after topical use of pure black cumin oil. Cases of contact dermatitis are reported to be caused by the use of cosmetics and perfumes that contain essential oils and are sold in markets as treatments for skin diseases. These cases were treated with topical corticosteroid application. The dose value that does not show any observable side effects in terms of reproductive toxicity in rats is IP 15 mg/kg. Since there is no information about its safety in oral and dermal exposure, it should not be used during pregnancy and breastfeeding. Yi et al. They suggested that TQ is safe to use during pregnancy due to its antiangiogenic effect. In humans and guinea pigs, it has been shown to inhibit uterine smooth muscle. Today, with the development of pharmaceutical technology, the use of herbal products used in traditional medicine has increased. Among these plant-based products, black cumin seed is also included. Black cumin seed and oil are also beneficial for health. Along with the active components it contains, many pharmacological effects such as anti-inflammatory, antioxidant, antimicrobial and anticancer have been determined by biological activity studies. Thymoquinone (T Q) is one of these active ingredients. Black cumin seed essential oil structure contains mainly TQ, dithymoquinone (DTQ), thymohydroquinone (THQ) and thymol (TMY) among the main pharmacologically active components. It is reported that TQ isolated from black seed seedlings has an anti-inflammatory effect by reducing the formation of reactive oxygen species (ROS) by inhibiting the cyclooxygenase (COX) and lipoxygenase (LOX) enzyme pathways that initiate the inflammation process. Treatment of eye-related diseases is usually carried out by topical application of medication. However, due to the unique structure of the eye, sufficient bioavailability cannot be achieved by applying topical medication to the eye. 11. Application of topical medication due to problems such as small eye volume and tear volume, reflexive blinking after medication is applied to the dead eye, excess tears passing into the nasolacrimal drainage, and absorption from the conjunctiva into the systemic circulation. The desired drug concentration in the eye cannot be achieved through this method. Since adequate bioavailability cannot be obtained with traditional topical dosage forms applied to the eye due to these problems, innovative ocular dosage forms have been developed in order to increase ocular bioavailability. These developed dosage forms have increased the contact time with the eye surface. It is applied to the patient. It is aimed to ensure patient compliance. In order for the topical dosage forms applied to the eye to be more effective, the contact time of the dosage form with the conjunctiva should be extended. It should be simple to apply for the patient. It should not cause irritation in the eye. It must have appropriate features. Microemulsions have been developed as new drug-containing systems to increase the ocular absorption of ophthalmic drugs. Most formulation strategies aim to maximize ocular drug permeability by prolonging drug residence time in the corneal and conjunctival area while simultaneously minimizing precomeal drug loss. Microemulsions are promising dosage forms for ophthalmic formulations as they have many advantages regarding ophthalmic application. Microemulsions are of particular interest in the delivery of hydrophobic drugs to the comea due to the possibility of loading the drug into the oil droplet. O/O (oil in water) microemulsions have been evaluated as a suitable system for ocular administration, dissolving poorly soluble drugs, increasing their absorption, and also prolonging the saline profile. Dry eye disease is an interpalpebral disease caused by tear deficiency or excessive tear evaporation. It was defined as a tear film disorder that causes ocular surface damage and symptoms of ocular discomfort in the field. 11. Today, this history has changed and dry eye is defined as a complex inflammatory syndrome of the lacrimal functional unit. There are few topical drugs available in the treatment of dry eye. Therefore, there is a need for anti-inflammatory and herbal-derived/innovative drug formulations for the treatment of dry eye disease. Ocular microemulsion formulations, which are innovative drug forms with thymoquinone (TQ) loading, are prepared. Products known in the art are products containing either artificial tears or cyclosporine. However, in this invention, black cumin oil? and use thymoquinone Jin Et 3. The primary aim of the invention is to benefit from the rich content of black cumin oil with anti-inflammatory properties. E. With the invention, black cumin essential oil contains thymoquinone, thymohydroquinone, dithymoquinone, thymol and side effects of these substances such as p-cymene, nigellidine and nigellisine. By selecting the oil phase of the ocular microemulsion system, which was developed by taking advantage of the anti-inflammatory properties of these main components with pharmacological properties such as black seed oil, and adding Thymoquinone, which has proven anti-inflammatory effects in the literature, as an active ingredient, an ocular eye drop was designed to provide the treatment of inflammation, which is a symptom of dry eye disease. With the invention, ocular microemulsion formulations were developed using black cumin oil for the first time. In the selection of black cumin oil, obtaining it cold pressed was determined as a prerequisite. Because it was proven by the literature that the total phenolic component content of black cumin oil obtained by the cold press method was high and low oxidative stability was observed. An ocular formulation was made for the first time, taking advantage of the anti-inflammatory properties of thymoquinone. In the invention, two different methods were used to prepare ocular microemulsion formulations. These are the phase titration method and the high-cycle homogenization method. While creating ocular microemulsion formulations, the surfactants they contain irritate the ocular surface. Therefore, the lyse surfactant It is necessary to develop the best formulation by using the minimum amount. Therefore, characterization and stability studies were carried out on the formulations developed using two different methods. As a result, the minimum surfactant content was determined in the ocular microemulsion formulations prepared according to the high-speed homogenization method and the ideal formulation was formed. According to the selected method, 30 microemulation formulations were prepared in a high speed homogenizer using black cumin oil surfactants, auxiliary surfactants and distilled water in certain proportions. Characterization studies Organoleptic evaluation for leaching, pH, viscosity, mass index, droplet. size, polydispersity index, zeta potential, density, phase separation and stability values were examined. The two best ocular microemulsion formulations were determined and Thymoquinone loading was done to the oil phase. The contents of the determined formulations were determined as F3-1-TQ Cremophor RH40 and 91.99% distilled water, together with the formulation codes: In vitro studies using ocular microemulsion formulations containing thymoquinone and dialysis membranes were carried out and pharmacokinetic evaluations were made in line with the data obtained in this study. Tween 20 and Cremophor RH40 were used as surfactants in the formulations. Cremophor RH40; Polyoxyl 40 hydrogenated castor oil and Tween 20; It is in the form of polyoxyethylene 20 sorbitan monolaurate. In addition to ocular microemulsion formulations, method validation of the Thymoquinone substance was carried out with an LC-MS/MS device for the first time in the literature. Solubility studies of Thymoquinone were performed for the first time in the literature on black cumin oil, Tween 80, Cremophor RH40, Cremophor ELP, propylene glycol, PEG 400 and glycer. Cytotoxicity studies on ARPE-19 cell line for the use of Thymoquinone in eye formulations were performed for the first time in the literature. Also containing Thymoquinone. The HET-CAM study was conducted for the first time in the literature to measure the irritation potential of the formulations on the eye surface. As a result, the irritation potential of the ocular microemulsion formulations was evaluated as "weak or slightly irritating" for F3-1-TQ and "non-irritating" for F7-9-TQ. Diagnostics of the Drawings Explaining the Invention Figure 1: Process Flow Diagram of the Formulation Prepared by the High Speed Homogenization Method Separation of the Invention [IJDAc [notation] The invention relates to effective eye drops in the treatment of dry eye disease containing black cumin oil and Thymoquinone (TQ). Bulus eye drops contain Dayrîla surfactant. The surfactant is Polyoxyethylene 20 sorbitan monolaurate (Tween 20) or Polyoxyl 40 hydrogenated castor oil (Cremophor RH40). Within the scope of the invention, two different eye drop formulations have been prepared. The first formulation contains 0.01% by weight Thymoquinone and 1.98% by weight black cumin oil. The formulation contains 0.01% by weight Thymoquinone, 2.00% by weight black cumin oil, 6.00% by weight Cremophor RH40 and 91.99% by weight distilled water. The surfactants selected in the invention are not irritating to the eyes, water By keeping the amount high, transparent emulsions with high patient compliance were obtained. The production cost was reduced by using a high amount of water in Ayr Ea. The amount of oil in it was sufficient to compensate for the lipid deficiency of the eye. The amount of thymoquinone was chosen to be effective but not toxic to the eye. Both formulations are suitable for application to the eye. Both formulations are suitable for application to the eye. The formulation has a long enough stay in the eye.HET-CAM test has shown that they are safe products for application to the eye. Kinetic studies showed that both formulations showed controlled and prolonged release. For the ocular microemulsion formulations developed by Hanm Fslt E. Invention, the oil phase forming the microemulsion was used as black seed oil obtained by cold pressing. Thus, black seed oil was used in the treatment of dry eye disease. The oil was extracted with anti-inflammatory active ingredients (e.g. TQ, DTQ, THQ, THY etc.) contained in the herb oil eluent. Substances that are used as surfactants forming the microemulsion and in eye formulations and that do not show irritating properties according to the literature were preferred. These are Cremophor RH40; Polyoxyl 40 hydrogenated castor oil and Tween 20; Polyoxyethylene 20 sorbitan monolaurate. At the same time, since it is known that the irritation potential is as low as 0 for the formulations developed for eye formulations, the lower the amount of surfactant, the lower the irritation potential, so stable formulations containing the least surfactant have been developed. For this purpose, formulations have been developed with two different methods. Distilled water was used as the water phase measurement. Thymoquinone was used as the active ingredient. Thymoquinone, which is also found in trace amounts in black cumin oil, was also included in the formulation by dissolving it in black seed oil (since it has lipophilic properties). Thus, by taking advantage of the anti-inflammatory properties of the formulation, it will help in the treatment of dry eye disease. a) Obtaining water phase 3 with surfactant and distilled water in a high speed homogenizer at 4000-16000 rpm, b) Obtaining oil phase 3 by mixing black seed oil and thymoquinone at 4000-16000 rpm. Obtaining ocular microemulsion by comparing with the corresponding value in the range of , Process names include Inlar Ii. Microemulsion pre-formulation studies were prepared as 10 mL using the high-speed homogenization method. Homogenizer conditions: 4000 rpm for 0 seconds, 30 formulations were prepared at room temperature3 (Figure 1). Selection of microemulsion formulations: The prepared pre-formulations were first observed and then placed in the stability cabinet (40 °C under moderate stability condition; 75% Humidity). The samples, which were kept for 3 months, were observed again and the droplet size, PDI (polydispersity index) and zeta potential values were checked with a zetasizer device. Microemulsion formulations were selected in line with the results obtained. Have you had dry eyes before, due to lack of tears or vaccination? It was diagnosed as a tear film disorder, resulting in the first symptoms of ocular surface damage in the interpalpebral area, caused by tear evaporation. Nowadays, this diagnosis has changed and dry eye is defined as a complex inflammatory syndrome of the lacrimal functional unit. Many effects of TQ have been observed, regardless of its pharmacological properties. Studies have been conducted in the fields of treatment such as TQ anti-inflammatory, antimicrobial, antibacterial, anticancer, nervous system, respiratory system, antidiabetic, antioxidant. In the invention, black cumin oil contains TQ, THQ, DTQ, TMY and these substances, which have pharmacological properties such as p-cymene, nigellidine and nigellisin. The following anti-inflammatory properties of the main components are mentioned in the literature. The cytotoxicity of the developed ocular microemulsion formulations was determined when applied to the ARPE-19 cell line and the formulation dose was determined accordingly. Separately, the potential for conjunctival irritation was determined with the HET-CAM Test. Ocular microemulsion formulations should be created using a high-speed homogenizer. In order to create ocular microemulsion formulations EiE, a high-speed homogenizer should be used to make the oil-in-water microemulsion more stable by using less surfactant and more oil phase. . In ARPE-19 cell line, Thymoquinone and ocular microemulsion formulations with and without Thymoquinone are used for lIl III cell survival. Determining the (IC50) value played an important role in determining the dosage amount. TQ and its formulations were studied for cytotoxicity using the MTT method on ARPE-19 cell line and IC50 values were determined. First, TQ was applied to ARPE-19 cells at different doses and at different times, and the cells were found in line with the data obtained. An increase in cell viability was observed at the 24th and 48th hours, depending on the low doses. watched very close to each other. It was observed that cell viability almost doubled at a dose of 37.5 µM TQ. At the 48th hour, it was observed that the effect of TQ on cell viability increased and it was determined that it increased cell reproduction. F3-1 and F 7-9 formulations, which do not contain TQ, were applied to ARPE-19 cells in different doses and at different times. In line with the data obtained, the cells 50% of them were found to be alive. It was determined that the effect of F3-1 formulation on cell viability was greater at 20 uL, 10 uL, 5 uL and 2.5 uL doses at the 24th hour, and cell viability decreased at the same doses at the 48th hour. However, at the lowest dose of 1.25 uL, a greater increase in cell viability was observed at the 48th hour. For the F7-9 formulation, its effect on cell viability increased at the 24th and 48th hours according to the decreasing dose. Cell survival at 24 hoursEIEgE48. It was determined that cell viability per hour was higher than all doses. Therefore, the 24th hour data were taken as basis for the F 7-9 formulation. As a result, the cell viability of both formulations was higher at the 24th hour. The F3-1 formulation showed slightly more cell viability than the F7-9 formulation at 20 uL and 10 uL doses; It is observed that the F7-9 formulation has higher cell viability at doses of 5 uL, 2.5 uL and 1.25 uL. The formulation with the highest ICso value can be preferred since it is F3-1. F3-1-TQ and F7-9-TQ formulations containing TQ were applied to ARPE-19 cells at different doses and at different times. According to the data obtained, 50% of the cells were found to be alive. It was found to be 45.15 uM per hour. The effect of the F 3-1-TQ formulation on cell viability was determined to decrease at higher doses. However, at the lowest dose, 41.5 µM cell survival]g348. More increases were seen per hour. For the F7-9-TQ formulation, cell survival was observed at 167.7 uM, depending on the decreasing dose, but at the 83.8 uM dose, cell viability decreased. Then, at the 41.9 uM dose, cell viability was observed again. It was determined that cell viability at the 24th hour was higher than the cell viability at the 48th hour compared to all doses. Therefore, the 24th hour data were taken as basis for the F7-9-TQ formulation. As a result, the cell survival at the 24th hour was higher in both formulations. E. F7-9-TQ can be preferred since the formulation with the highest IC50 value is F7-9-TQ. Cytotoxicity studies were conducted to determine the cell viability and IC50 value of TQ-loaded formulations, and the formulation that gave the best IC50 value was found to be the F 7-9-TQ formulation. The reason for this is that when we examined the results of the F 7-9-TQ formulation in the salen study, it provided more controlled salience than the F3-1-TQ formulation, and accordingly, the droplet size and PDI values were more stable. At the same time, its viscosity was higher and the residence time in the environment could be longer. With these data, the interpretation that it could produce a linearly prolonged release following the first order and Korsmeyer-Peppas models as a result of the kinetic massage was evaluated. Therefore, when the in vitro results of F7-9-TQ were evaluated, it was proven that cell viability was increased by making a slow, effective and controlled release in cell lines at first, but this effect decreased over time and cell viability decreased. Determination of irritation potential of formulations using Ibrak HET-CAM test using embryonated chicken eggs Hen's Egg Test on Caryoallantoic Membrane (CAM) is another alternative method for animal experiments for the analysis of as EldiBD and/or severe ocular irritants using the karyoallantoic membrane of embryonated chicken eggs. This test is assesses damage to this membrane to determine potential irritation of the conjunctiva. It is assumed that the acute effects of the test substance on the small vasculature and proteins of the soft tissue of the membrane are similar to the effects caused by the substances in the eyes of rabbits. TQ-loaded ocular microemulsion formulations were tested using this method and In comparison with the results obtained with 0.9% NaCl solution as control [[hstEEldüF 3-1-TQ and F 7-9-TQ formulations, bleeding, vascular lysis and pEElt formation were observed. When the F3-1-TQ formulation was applied to the chorioallantoic membrane, 266 Vascular lysis was observed after .4 seconds and the IS was found to be 0.346. When IS was evaluated, the formulation was not irritating. When the EsaptandD F7-9-TQ formulation was applied to the chorioallantoic membrane, no bleeding, vascular lysis or clotting was observed, and it did not cause any irritation. As a result, the prepared TQ-loaded ocular microemulsion formulations were shown to be safe for ocular application.TR

Claims (1)

1.CLAIMS. It is an effective eye drop in the treatment of dry eye disease and its features are; It contains black cumin oil and Thymoquinone (TQ). . Ealup feature of eye drops in accordance with Claim 1; It contains surfactant. . It is an eye drop that complies with claim 2 and its feature is; The surfactant is Polyoxyethylene 20 Surbitan monolaurate (Tween 20) or Polyoxyl 40 hydrogenated castor oil (Cremophor RH40) 20) and agElEça contains 95.04% distilled water. It is an eye drop according to claim 3 and its feature is; agElEça 0.01% Eda Thymoquinone, ag ElEkça castor oil (Cremophor RH40) and agElEça 91.99% ratio. Eda contains distilled water. It is an eye drop EE preparation method and its feature is; ) Obtaining the oil phase by mixing black cumin oil and thymoquinone in the speed range of 4000-16000 rpm. s Contains E. TR