Patent Application
20230211362
The drug soaking method relies on passive diffusion of a drug into the lens materials. This process is simple, but wasteful. The capacity is low and loading and release from soaked lenses are highly uncontrolled leading to rapid release kinetics and a varying rate of release. The molecular imprinting method allows the incorporation of drug molecules into the contact lenses polymeric material, although typically only of one drug at a time. This drug loading method could extend the release of drug to weeks. However due to differences in the chemistry of each ocular drug, each contact lens material would be different, so regulatory approval would be needed for each different drug to reach the market. Using the layering technique, the manufacture process contains multiple steps and is highly complex. For example, latanoprost DECLs that have been made in this way have an encapsulated drug-polymer film within the contact lens hydrogel [14]. The drug-polymer film has to be placed within the contact lens hydrogel and the dried hydrogel then has to be precisely lathed so that the drug-polymer is encapsulated by the contact lens hydrogel.
Additive coating contact lenses using drug-containing materials has also been proposed. The coating concept is simple and has the advantage of potentially being an additive step that can be implemented into the manufacturing process of the contact lenses. Spin coating, which involve centrifugal coverage of the entire lens, has been suggested and previously tested by us as a coating technique but suffers from the disadvantages of high drug wastage as the drug-polymer is lost from the edge of the lenses, which leads to a low level of drug loading accuracy. In addition, if the drug-loaded film has any opacity after hydration, the lenses will lose their clarity and affect vision. Conventional electrospinning and electrospray [e.g. see US 2004/0170666 A1; U.S. Pat. No. 9,956,168 B2, US 2005/0067287 A1; US 2007/0157880 A1; EP 2 529 761 E1; EP 1988941] have also been reported for coating of hydrogel surface and contact lenses.
The difference between the invention described in this disclosure and the above-mentioned reference is that conventional electrospray and electrospinning require the pumping mechanism (i.e. syringe pumps) to continuously feed the solution to the nozzle for deposition, as illustrated in
Inkjet printing is an established drop-on-demand technique that can deposit low viscosity liquid formulations (<30 cp) with typical drop volumes of 1-5 pL and a resulting feature size of 20-50 micron [16]. Although the inkjet method may be used to selectively print on certain portions of the lens surface, it will suffer significant difficulties in coating the hemispherical geometry of a contact lens due to highly constrained printable liquid properties. Inkjet requires low-viscosity, high-solvent content which would lead to long drying times for printed material and issues of flow and spreading of the drug coating post-deposition due to the curved substrate. This could lead to unwanted coating within the visual axis of the lens. Another drawback of inkjet is that the low printing resolution results in a relatively large droplet volume which will produce a sudden step at the edge of the deposited drug film. This relatively prominent feature could be problematic and cause increased friction due to blinking and discomfort to the wearer.
The present inventors wished to develop a method that is an alternative to, ideally an improvement upon, the various prior art methods of applying an active compound to a contact lens or other ocular prosthetic device, and which addresses one or more of the problems associated with the prior art techniques.
In a first aspect, there is provided a method of coating an ocular prosthetic device, the method comprising
nanoelectrospraying droplets comprising an active ingredient and/or a carrier species onto a surface of the ocular prosthetic device in a predetermined pattern, the nanoelectrospraying involving controlling the flow rate of the droplets from a nozzle of the nanoelectrospraying equipment by controlling the voltage between the nozzle and the ocular prosthetic device.
In a second aspect, there is provided an ocular prosthetic device formable according to the method of the first aspect. Any liquid in the droplets will typically have been removed (e.g. evaporated) to leave a layer comprising the active ingredient and/or carrier species on the surface of the prosthetic device.
In a third aspect, there is provided a method of coating a surface of a prosthetic device, the method comprising
nanoelectrospraying droplets comprising an active ingredient and/or a carrier species onto the surface in a predetermined pattern, the nanoelectrospraying involving controlling the flow rate of the droplets from a nozzle of the nanoelectrospraying equipment by controlling the voltage between the nozzle and the prosthetic device. The surface may be of any curvature, e.g. curved (which may be selected from convex and concave) or flat. The surface may be electrically conducting, insulating or electrically semi-conducting. Preferably, the surface is a curved and/or electrically conductive surface.
In a fourth aspect, there is provided a prosthetic device formable according to the method of the third aspect. The surface onto which the droplets have been nanoelectrosprayed may be of any curvature, e.g. curved or flat. The surface onto which the droplets have been nanoelectrosprayed may be electrically conducting, insulating or electrically semi-conducting. The prosthetic device may be a device with a curved and/or electrically conductive surface (and which is the surface on with the droplets are nanoelectrosprayed), such as a stent. Any liquid in the droplets will typically have been removed (e.g. evaporated) to leave a layer comprising the active ingredient and/or carrier species on the surface of the prosthetic device.
The present invention is a unique liquid deposition method involving nanoelectrospray (nES) onto ocular and other prosthetic devices, which allows high precision control to deposit controlled mass of materials onto surfaces, e.g. electrically conductive surfaces, of the ocular prosthetic device or other prosthetic device with pre-designed pattern at pre-determined location. In more detail, the method allows precise deposition of a desired amount of material onto a surface, which may be an electrically conductive surface (such as a wet surface or a metal surface), to form a coating layer on the surface. This allows the formation of the pre-designed coating pattern of the materials that have been deposited onto the conductive surface. The coat material and the pattern in combination carry functionality such as controlled delivery of drug at specific site of the body. Examples of such include a ring-shaped drug layer at the edge of contact lenses to allow targeted drug delivery to the eye at a controlled rate. It may also be applied to other prosthetics, e.g. by applying a drug-containing coating onto the highly curved surfaces of carotid artery stents. Depending on the processing parameters and the material properties, the deposited coat layer can be either a smooth and continuous layer of materials with no pinhole or a layer formed by semi-fused nanoparticles. The method can also generate multiple-layer deposition in which the layers overlay with each other. The dimension of the area covered by the deposition is tuneable by the nES operational parameters.
The method has distinct difference from conventional electrospray method in terms of the method used to control the liquid jetting for deposition. This leads to much narrower and streamlined spray of the material leading to more precise and selective deposition location of the materials in comparison to conventional electrospray. It does not allow deposition of a material onto a large non-selective area.
The advantages can include:
As mentioned above, some prior art techniques have two significant disadvantages: 1) waste of active ingredient and no control of the amount of the materials that are deposited on the lens; 2) the full coverage of the lens surface leads to the loss of clarity of vision zone of lens, as shown in the
Herein we describe the use of additive deposition technology using nanoelectrospray (nES), to deposit the drug formulation on selected areas of an ocular prosthetic device, e.g. a contact lens. nES is a form of ES operation where the fluid flow rate is controlled by the applied voltage and nES is achieved without the need for pumps or valves [17, 18]. In nES, for a particular applied voltage, the droplet size and volumetric flow rate will depend on (as well as the applied voltage), the liquid properties such as conductivity, surface tension and viscosity and on nozzle geometry. It has been shown to be a versatile tool for numerous applications but most notably, it has been proven as a benign technique for the ‘soft’ ionisation of biomolecules, most commonly used in mass spectrometry. At present, the most studied ES form is the cone-jet regime, wherein the balance between electrostatic stresses and surface tension creates a Taylor cone and from the apex of which a liquid jet is emitted [17].
Conventional electrospray (ES) for additive printing can generate liquid droplets of microns to nanometer size by electrostatic force to form a film coat. However as demonstrated in
Therefore, nES can be used to coat the ocular prosthetic device, e.g. a contact lens, with a continuous coating in a certain desired pattern, e.g. to coat a fraction of the surface area of the ocular prosthetic device, e.g. a contact lens, allowing for active ingredient for delivery to the eye at from the coated area(s). nES can generate liquid droplets 10s of microns to nanometer size and with the use of appropriate formulation and processing parameters, the technique can deposit semi-dried droplets to form a film coat on the ocular prosthetic device's, e.g. a contact lens', surface. The film coat may contain excipients including a polymer and an active compound. This ability to print higher drug and polymer loadings with lower solvent content, when compared to inkjet, would result in minimal flow of deposited material prior to drying with the ability to design formulations to fuse or dry almost instantaneously on landing, of particular benefit when addressing wet contact lenses. Regarding patient comfort, due to the high-resolution and low-volume capabilities the film coating profile can be controlled to result in a shallower/smoother edge profile and increased patient comfort due to the deposition of nm to micron-sized droplets within the spray.
More importantly, instead of printing the formulation to cover the entire surface of the ocular prosthetic device, e.g. a contact lens, nES is capable of selectively coating a certain proportion of ocular prosthetic device's, e.g. a contact lens', surface to achieve the patterned printing (as seen in
nES allows the controlled ejection and deposition of minute volumes (sub-picolitre) of highly viscous liquid (up to 1000 cp, required for drug formulation) which is not possible by conventional inkjet printing. The nES printing technique allows excellent reproducibility with volume variation demonstrated to be less than 3% to enable high precision drug dosing.
In a first aspect, there is provided a method of coating an ocular prosthetic device, the method comprising
nanoelectrospraying droplets comprising an active ingredient and/or a carrier species onto the surface of the ocular prosthetic device in a predetermined pattern, the nanoelectrospraying involving controlling the flow rate of the droplets from a nozzle of the nanoelectrospraying equipment by controlling the voltage between the nozzle and the ocular prosthetic device.
Nanoelectrospraying involves controlling the flow rate of the droplets from a nozzle of the nanoelectrospraying equipment by controlling the voltage between the nozzle and the ocular prosthetic device. The ocular prosthetic device may be placed on a surface, e.g. an electrically conductive surface. The electrically conductive surface may comprise an electrically conductive material, which may be selected from a metal, carbon, and organic or inorganic electrically conducting material. The metal may be any metal, including any transition metal, such as, for example, copper, brass, zinc, iron, steel, gold, silver. The surface may comprise a transparent, electrically conducting material, such as a transparent conducting oxide, which may be, for example, selected from an indium tin oxide, which may be doped, for example with fluorine. doped zinc oxide, such as indium zinc oxide, aluminium zinc oxide. The organic electrically conducting material may comprise an electrically conductive polymer, for example a polymer selected from poly(3,4-ethylenedioxythiophene) (PEDOT), poly(3,4-ethylenedioxythiophene) PEDOT: poly(styrene sulfonate) PSS (i.e. the material PEDOT:PSS), poly(4,4-dioctyl cyclopentadithiophene). The carbon may be any form of electrically conducting carbon, which may comprise, for example graphite. In an embodiment, the carbon comprises a carbon nanotube, and the surface comprising the carbon nanotube may be transparent. An advantage of the surface being transparent is that it allows light transmission from below the ocular prosthetic device, which is typically transparent, which allows more accurate analysis of the ocular prosthetic device and any materials applied to it during and after coating process.
Controlling the flow rate of droplets may be carried out by setting a nozzle to substrate distance (the substrate being the ocular prosthetic device), and then determining the minimum (onset) voltage between the nozzle and the ocular prosthetic device for a droplet to eject from the nozzle (Von). To eject droplets from the nozzle, the voltage may then be altered from a voltage V1, which is below Von, to V2, which is above Von. V1, may be ground (0V) or a voltage less than Von. V2-V1 may be of from about 0.05 kV to 5 kV, optionally 0.05 kV to 4 kV, optionally 0.05 kV to 3 kV, optionally 0.05 kV to 2 kV, optionally 0.05 kV to 1 kV, optionally 0.05 kV to 0.5 kV, optionally about 0.1 to 0.4 kV. V2 will depend on a variety of factors, including the nature of the droplets and the distance from nozzle to substrate. However, in general terms, V2 may be from 1.5 kV to 3 kV. The voltage may be circulated between V1 and V2 with a particular frequency. The frequency may be from 0.1 Hz to 20 kHz, optionally 0.5 Hz to 15 kHz, optionally 1 Hz to 10 kHz, optionally optionally 1 Hz to 5 kHz, optionally 1 Hz to 1 kHz, preferably optionally 10 Hz to 1000 Hz is preferably 1 to 20 Hz, preferably 5 to 15 Hz. While the nozzle is not spraying (e.g. when the voltage is at V1), the ocular prosthetic device may be moved relative to the nozzle, and then the ocular prosthetic device held stationery relative to the nozzle while spraying, and this may be repeated, to allow a build-up of a pattern on the surface. In alternative embodiments, there may be movement of the ocular prosthetic device during spraying, to allow build-up of a pattern.
Controlling the voltage between the nozzle and the ocular prosthetic device may mean varying the voltage to induce droplet formation. In some examples, varying the voltage may comprise changing the voltage from a first voltage to a second (higher) voltage, and the nozzle may eject a droplet or spray of droplets when the voltage is the second (higher) voltage but not when the voltage is the first (lower) voltage.
The voltage may be a DC voltage or an AC voltage. The voltage may be a pulsed voltage. The variation of the AC voltage may be sinusoidal voltage or a square wave voltage, optionally with a constant frequency switch between V1 and V2. This has the effect of switching the spray on and off and slowing the volumetric throughput, which promotes the evaporation of any liquid in the droplets after deposition, minimising flow of the droplets after they have landed on the surface.
In some embodiments a backpressure may be applied to the liquid in the nozzle during nanoelectrospraying. This may be from a chamber in fluid connect from a nozzle that is pressurized. In an embodiment, the chamber may be a syringe or other pressurisable chamber in fluid connection with the nozzle. A backpressure of from 0.01 bar to 1 bar, optionally 0.01 bar to 0.1 bar may be applied. This may assist the ejection of the droplets. The pressure may be raised and lowered, e.g. in a cycle, during the nanoelectrospraying, optionally in synchronization with any cycle of the voltage variation.
The distance from the nozzle to the ocular prosthetic device during the nanoelectrospraying is preferably from 0.1 mm to 5 mm, preferably 0.5 mm to 4 mm, preferably 0.5 mm to 3 mm, preferably 0.5 mm to 2 mm, preferably about 1 mm. The smaller distance from the nozzle has been found to lead to a smaller surface area of the lens being coated with a spray of droplets. This leads to finer control of the pattern being produced on the ocular prosthetic device. It was found, for example, that a width of coating of less than 3 mm could be achieved with a nozzle to ocular prosthetic device distance of 3 mm, and that a width of coating of less than 1 mm could be achieved with a nozzle to ocular prosthetic device distance of 1 mm.
The distance from the nozzle to the position on the ocular prosthetic device at which a droplet (or the center of a spray of droplets) will be deposited (nozzle to substrate distance, NSD) during the nanoelectrospraying is preferably 5 mm or less, for example, 4.5 mm or less, 4 mm or less, 3.5 mm or less, 3 mm or less, 2.5 mm or less, 2 mm or less, 1.5 mm or less, 1 mm or less, 0.5 mm or less, or 0.1 mm or less. The distance from the nozzle to the ocular prosthetic device during the nanoelectrospraying is preferably from 0.1 mm to 4.5 mm, for example, 0.5 mm to 4 mm, 1 mm to 3.5 mm, 1 mm to 3 mm, 1.5 mm to 2.5 mm, 0.1 mm to 2 mm. In some examples, it was found that the width of coating increases linearly with the distance from the nozzle to the ocular prosthetic device and/or the thickness of the coating decreases linearly with the distance from the nozzle to the ocular prosthetic device. In some examples, the ocular prosthetic device is a contact lens and the NSD is the distance from the nozzle to a point on the surface of the contact lens where a droplet will be deposited (which may correspond to the center of the deposited active ingredient and/or carrier species on the surface of the contact lens).
The nozzle may have an inner diameter of 250 μm or less, optionally 200 μm or less, optionally 150 μm or less, optionally 100 μm or less. The nozzle may have an inner diameter of from 10 μm to 250 μm, optionally 10 μm to 200 μm, optionally 50 μm to 200 μm, optionally 50 μm to 150 μm, optionally 75 μm to 125 μm, optionally about 100 μm.
The flow rate of droplets from the nozzle may be from 0.1 nL/min to 5000 nL/min, optionally from 1 nL/min to 1000 nL/min, optionally from 1 nL/min to 100 nL/min.
During the nanoelectrospraying, the substrate (i.e., the ocular prothetic device) and the nozzle of the nanoelectrospraying equipment may move relative to each other. In some examples, the nozzle is stationary and the substrate is in motion. In some examples, the substrate is stationary and the nozzle is in motion. In some examples, both the nozzle and the substrate are in motion, for example, in opposite directions. The relative movement may be at a speed of at least about 0.1 mm/s, for example, at least about 0.5 mm/s, at least about 1 mm/s, at least about 1.5 mm/s, at least about 2 mm/s, at least about 2.5 mm/s, at least about 3 mm/s, at least about 3.5 mm/s, at least about 4 mm/s, at least about 4.5 mm/s, at least about 5 mm/s, at least about 5.5 mm/s, at least about 6 mm/s, at least about 6.5 mm/s, at least about 7 mm/s, at least about 7.5 mm/s, at least about 8 mm/s, at least about 8.5 mm/s, at least about 9 mm/s, at least about 9.5 mm/s, at least about 10 mm/s, at least about 15 mm/s, at least about 20 mm/s, at least about 25 mm/s, at least about 30 mm/s, at least about 35 mm/s, at least about 40 mm/s, at least about 45 mm/s, or at least about 50 mm/s. In some examples, the relative movement may be at a speed of up to about 50 mm/s, for example, up to about 45 mm/s, up to about 40 mm/s, up to about 35 mm/s, up to about 30 mm/s, up to about 25 mm/s, up to about 20 mm/s, up to about 15 mm/s, up to about 10 mm/s, up to about 9.5 mm/s, up to about 9 mm/s, up to about 8.5 mm/s, up to about 8 mm/s, up to about 7.5 mm/s, up to about 7 mm/s, up to about 6.5 mm/s, up to about 6 mm/s, up to about 5.5 mm/s, up to about 5 mm/s, up to about 4.5 mm/s, up to about 4 mm/s, up to about 3.5 mm/s, up to about 3 mm/s, up to about 2.5 mm/s, up to about 2 mm/s, up to about 1.5 mm/s, up to about 1 mm/s, up to about 0.5 mm/s, or up to about 0.1 mm/s. In some examples, the relative movement may be at a speed of from about 0.1 mm/s to about 50 mm/s, for example, about 0.5 mm/s to about 45 mm/s, about 1 mm/s to about 40 mm/s, about 1.5 mm/s to about 35 mm/s, about 2 mm/s to about 30 mm/s, about 2.5 mm/s to about 25 mm/s, about 3 mm/s to about 20 mm/s, about 3.5 mm/s to about 15 mm/s, about 4 mm/s to about 10 mm/s, about 4.5 mm/s to about 9.5 mm/s, about 5 mm/s to about 9 mm/s, about 5.5 mm/s to about 8.5 mm/s, about 6 mm/s to about 8 mm/s, about 6.5 mm/s to about 7.5 mm/s, or about 7 mm/s to about 50 mm/s. In some examples, it was found that the width of the coating was not significantly affected by the speed at which the substrate and/or the nozzle moved and/or that the thickness of the coating decreased (for a fixed number of layers, e.g., rotations of the nozzle or substrate) with the speed at which the substrate and/or the nozzle moved.
The droplets will typically include a liquid in which the active ingredient and/or the carrier species is dispersed, e.g. dissolved. A plurality of active ingredients and/or a plurality of carrier species may be present in the droplets, and then in the coating on the ocular prosthetic device. The liquid may be selected from a polar solvent and a non-polar solvent. The solvent may be a protic polar solvent or a non-polar protic solvent. The polar protic solvent may be selected from water, an alkanol, e.g. n-butanol, isopropanol, n-propanol, ethanol, and a carboxylic acid, such as formic acid or acetic acid. The polar non-protic solvent may be a solvent such a solvent selected from dichloromethane, tetrahydrofuran, ethylacetate, acetone, dimethylformamide, acetonitrile and dimethyl sulphoxide.
The ocular prosthetic device may have a concave surface and/or a convex surface and the droplets and the active ingredient and/or the carrier species may be applied to the concave surface and/or convex surface.
The carrier species may be a polymer, preferably a biocompatible polymer. The carrier species may be a polymer selected from sodium polystyrene sulfonate (PSS), polyethers, such as a polyethylene oxide (PEO), polyoxyethylene glycol or polyethylene glycol (PEG), polyethylene imine (PEI), a biodegradable polymer such as a polylactic acid, e.g poly-d,I-lactic acid, polycaprolactone, polyglycolic acid, poly(lactic-co-glycolic acid) (PLGA), zein, poly(I-lactide-co-ε-caprolactone) (PLCL), hyaluronic acid, polymethylsilsesquioxane (PMSQ) and copolymers, derivatives, and mixtures thereof. Other polymers that may be used include those well known to those of skill in the art to be used in cell cultures, implants, regenerative, therapeutic, and pharmaceutical compositions. One such example is polyvinylpyrrolidone (PVP).
The polymer may be a water-soluble and/or hydrophilic polymer, which may be selected from biocompatible polymers, including, but not limited to, cellulose ether polymers, including those selected from the group consisting of hydroxyl alkyl cellulose, including hydroxypropyl methyl cellulose (HPMC), hydroxypropyl cellulose (HPC), hydroxyethyl cellulose (HEC), methyl cellulose (MC), carboxymethyl cellulose (CMC), and mixtures thereof.
The polymer may also be selected from polyvinylpyrrolidone, vinyl acetate, polyvinylpyrrolidone-vinyl acetate copolymers, polyvinyl alcohol (PVA), acrylates and polyacrylic acid (PAA), including polyacrylate polymer, vinylcaprolactam/sodium acrylate polymers, methacrylates, poly(acryl amide-co-acrylic acid) (PAAm-co-AA), vinyl acetate and crotonic acid copolymers, polyacrylamide, polyethylene phosphonate, polybutene phosphonate, polystyrene, polyvinylphosphonates, polyalkylenes, and carboxy vinyl polymer. The multiphasic fiber compositions may comprise derivatives, copolymers, and further combinations of such polymers, as well.
The polymer may be selected from water insoluble or hydrophobic polymers including, but not limited to, cellulose acetate, cellulose nitrate, ethylene-vinyl acetate copolymers, vinyl acetate homopolymer, ethyl cellulose, butyl cellulose, isopropyl cellulose, shellac, hydrophobic silicone polymer (e.g., dimethylsilicone), polymethyl methacrylate (PMMA), cellulose acetate phthalate and natural or synthetic rubber; siloxanes, such as polydimethylsiloxane (PMDS), cellulose, polyethylene, polypropylene, polyesters, polyurethane and nylon, including copolymers, derivatives, and combinations thereof.
In a preferred embodiment, the carrier species is a polymer selected from PLGA, zein, PLCL, and hyaluronic acid.
Poly(lactide-co-caprolactone) (PLCL) is a co-polymer of poly(lactic acid) and polycaprolactone. The co-polymer is biocompatible and has biodegradable characteristics. They show high degradation and hydrophobicity as well as good miscibility with other polymers and the permeability is excellent for a drug-carrying polymer. These aliphatic polymers are approved by the FDA and as such are extensively used in the pharmaceutical sector.
Poly (Lactic-co-Glycolic Acid) (PLGA) 50:50, with acidic terminal group (used in the Examples). PLGA, approved by the FDA, is an extensively used polymer in the pharmaceutical industry as it has characteristics favourable to releasing drug in a controlled manner. The polymer is a synthetic biodegradable and non-immunogenic product that can have its physicochemical properties altered by changing the ratio of PGA and LA during its production. PGA is highly crystalline material and hydrophilic but when combined with LA becomes a polymer with a longer degradation time due to higher hydrophobicity and less swelling capacity.
Zein is a class of protein found in maize. The commercial source of zein is a fine yellow powder which partially soluble in water alone. It solubilizes in a mixture of ethanol (70-80% w/w) and water (30-20% w/w) due to it amphiphilic nature. It is a biodegradable and biocompatible material.
The carrier species, e.g. polymer, may be present in the droplets in a concentration of from 0.1 mg/ml to 100 mg/ml, optionally 1 mg/ml to 100 mg/ml, optionally from 5 mg/ml to 75 mg/ml, optionally from 10 mg/ml to 75 mg/ml, optionally from 10 mg/ml to 50 mg/ml, optionally from 50 mg/ml to 75 mg/ml.
The active ingredient may be or comprise a pharmaceutically active ingredient, optionally two or more pharmaceutically active ingredients. The pharmaceutically active ingredient may be selected from a small molecule drug, proteins, nucleic acids, polysaccharides, and biologics. A biologic may be selected from a vaccine, blood, blood components, cells, genes, tissues and recombinant proteins. A biologic may be selected from a protein that can control the action of other proteins and cellular processes, a gene that control the production of proteins, a hormone, which may be a modified hormone, and a cell that produces one or more substances that suppress or activate components of the immune system. A biologic may be referred to as a biologic response modifier. The pharmaceutically active ingredient may be any agent capable of providing a therapeutic or a prophylactic benefit to a human or animal wearing an ocular prosthetic device having the pharmaceutically active ingredient thereon or therein. In an embodiment, the pharmaceutically active ingredient is effective for treating diseases and disorders of the eye or an ocular disease or disorder. In non-limiting, exemplary embodiments, the pharmaceutically active ingredient is selected from an anti-infective agent (e.g., an antibiotic or antifungal agent), an anesthetic agent, an anti-VEGF agent, an anti-inflammatory agent, a biological agent (i.e. a biologic, as described above. The biological agent may be or comprise a macromolecular molecule, including, but not limited to, RNA, DNA, antibodies or fragments thereof, peptides, proteins and carbohydrates), an intraocular pressure reducing agent (i.e., a glaucoma drug), or a combination thereof. Non-limiting examples of drugs are provided below. The ocular disease or disorder may be selected from disorders such as post-surgical and post-trauma pain, glaucoma, uveitis, and hay fever allergy and dacrocystitus.
The active ingredient may be selected from growth factors, angiogenic agents, anti-inflammatory agents, anti-infective agents such as antibacterial agents, antiviral agents, antifungal agents, and agents that inhibit protozoan infections, antineoplastic agents, anaesthetics, anti-cancer compositions, autonomic agents, steroids (e.g., corticosteroids), non-steroidal anti-inflammatory drugs (NSAIDs), antihistamines, mast-cell stabilizers, immunosuppressive agents, antimitotic agents, or other drug. In some embodiments, the active agent may be a wetting agent, surfactant, ocular demulcent, electrolyte, buffer, or preservative. In some cases, an active agent may improve a wearer's comfort when wearing a ocular prosthetic device, e.g. contact lens.
The antibacterial agent may be selected from bacitracin, chloramphenicol, ciprofloxacin, erythromycin, moxifloxacin, gatifloxacin, gentamicin, levofloxacin, sulfacetamide, polymyxin B, vancomycin, tobramycin, or a combination thereof.
The antiviral agents may be selected from trifluridine, vidarabine, acyclovir, valacyclovir, famciclovir, foscarnet, ganciclovir, formivirsen, and cidofovir.
The antifungal agents may be selected from amphotericin B, natamycin, fluconazole, itraconazole, ketoconazole, and miconazole.
The antiprotozoal agents may be selected from polymyxin B, neomycin, clotrimazole, miconazole, ketoconazole, propamidine, polyhexamethylene biguanide, chlorhexidine, itraconazole.
The anesthetic agents may be selected from an aminoamide and an aminoester. The aminoamide may be selected from lidocaine, prilocalne, mepivacaine and ropivacaine.
The aminoesters may be selected from benzocaine, procaine, proparacaine, and tetracaine.
The autonomic agents may be selected from acetylcholine, carbachol, pilocarpine, physostigmine, echothiophate, atropine, scopolamine, homatropine, cyclopentolate, tropicamide, dipivefrin, epinephrine, phenylephrine, apraclonidine, brimonidine, cocaine, hydroxyamphetamine, naphazoline, tatrahydrozoline, dapiprazole, betaxolol, carteolol, levobunolol, metipranolol, and timolol.
The anti-inflammatory agents may be selected from a non-steroidal anti-inflammatory agent, and a steroidal anti-inflammatory agent. Non-limiting examples include glucocorticoids (e.g., dexamethasone, prednisolone, fluorometholone, loteprednol, medrysone, and rimexolone) and NSAIDS (e.g., diclofenac, flurbiprofen, ketorolac, bromfenac, and nepafenac).
The pharmaceutically active ingredient may be selected from ophthalmic antihistamines, ophthalmic prostaglandin analogs, ophthalmic α-adrenoreceptor agonists, ophthalmic p-adrenoreceptor agonists, ophthalmic carbonic anhydrase inhibitors and ophthalmic parasympathomimetic agents. The ophthalmic antihistamines may be selected from alcaftadine, azelastine, bepotastine, cetirizine, epinastine, ketotifen, and olopatadine. The ophthalmic prostaglandin analogs may be selected from latanoprost, travoprost, bimatoprost, tafluprost, and unoprostone isopropyl. The ophthalmic α-adrenoreceptor agonists may be selected from brimonidine and apraclonidine. The ophthalmic p-adrenoreceptor agonists may be selected from timolol, betaxolol, carteolol and levobunolol. The ophthalmic carbonic anhydrase inhibitors may be selected from acetazolamide, dorzolamide and brinzolamide. The ophthalmic parasympathomimetic agents may be selected from carbachol and pilocarpine.
In an embodiment, the active ingredient is selected from ketotifen fumarate, timolol and latanoprost and sodium hyaluronate.
Ketotifen fumarate has a molecular weight of 250.50, pKa 12.3 (acid), Log P 3.4. It is a second-generation antihistamine drug for treating seasonal allergic conjunctivitis (SAC). The physicochemical characteristics of KF suggest that this drug is slightly soluble in aqueous solutions (7.87 μg/ml) and as such will dissociate from a polymer layer and move into the aqueous environment via diffusion.
Timolol is a nonselective beta-adrenergic antagonist given in an eye drop solution to reduce intraocular pressure, or pressure in the eyes.
Latanoprost is a prostaglandin F2α analogue, is a selective prostanoid FP receptor agonist which reduces the intraocular pressure by increasing the outflow of aqueous humour.
Sodium Hyaluronate (HA) is a naturally occurring anionic non-sulfated glucosaaminoglycon linear polysaccharide which has been used as an artificial tear solution. Due to its hygroscopic nature, it provides treatment for ocular discomfort and dry eye syndrome. It has an important role in structural and biological roles in animal tissues. The long polysaccharide chain provides the viscoelasticity which enhances the tear stability.
The active ingredient may be a compound for increasing the comfort of the wearer and/or improving the hydration of the ocular prosthetic device, e.g. contact lens, and eye in contact with the ocular prosthetic device, such as hyaluronic acid.
The pharmaceutically active agent may be present in the droplets in a concentration of from 0.1 mg/ml to 50 mg/ml, optionally from 0.1 mg/ml to 20 mg/ml, optionally from 0.1 mg/ml to 10 mg/ml.
The amount of active ingredient deposited on the ocular prosthetic device may be an amount suitable for it to carry our its function, e.g. therapeutic or prophylactic function if the active is a pharmaceutical active ingredient. The active ingredient may, for example, be present on the ocular prosthetic device in an amount of from 0.1 μg to 100 μg, optionally from 1 μg to 50 μg.
The active ingredient may be a colouring agent for imparting a colour to the ocular prosthetic device, e.g. contact lens, for cosmetic purposes. The colouring agent may be selected from a solubilized leuco sulfate ester of 7,16-Dichloro-6,15-dihydro-5,9,14,18-anthrazinetetrone (C.I. Vat Blue 6), 16,23-Dihydrodinaphtho[2,3-a:2′,3′-i]naphth[2′, 3′:6:7]indole[2,3-c]carbazole-5,10,15,17,22,24-hexone (C.I. Vat Brown 1), N,N′-(9,10-Dihydro-9,10-dioxo-1,5-anthracenediyl)bisbenzamide (C.I. Vat Yellow 3), 16,17-Dimethoxydinaphtho[1,2,3-cd:3′,2′,1′-1n]perylene-5,10-dione (C.I. Vat Green 1), 6-Ethoxy-2-(6-ethoxy-3-oxobenzo[b]thien-2(3H)-ylidene)benzo[b]thiophen-3(2H)-one (C.I. Vat Orange 5).
The droplets may further include an inorganic salt to increase their conductivity. The conductivity of the droplet, measured in a sample of a bulk of the liquid of the droplets, may be from 0 to 1 μS/m, optionally from 0.1 μS to 10 mS/m, optionally from 0.1 μS to 100 mS/m, optionally from 0.1 μS to 10 mS/m, optionally from 0 to 200 μS/m, optionally from 5 μS to 100 μS/m, optionally from optionally from 10 μS to 30 μS/m. The inorganic salt may, for example, be an ammonium salt, such as a tetraalkylammonium halide, such as tetrabutylammonium bromide (TBAB) or an alkali metal salt, such as an alkali metal halide, such as sodium chloride.
The droplets may have a dynamic viscosity, measured at 20° C. and standard pressure, of from 1 cP to 10000 cP, optionally 1 cP to 1000 cP, optionally 20 cP to 1000 cP, optionally from 50 cP to 1000 cP, optionally from 100 to 1000 cP. The dynamic viscosity may be measured using any standard technique, e.g. using a capillary viscometer, a rotational viscometer (e.g. using ISO 3218.2) or a rolling ball viscometer.
The droplets, and hence the active ingredient and/or carrier species may be coated on the ocular prosthetic device in a predetermined pattern. The pattern may be an array of discrete dots, of any shape, lines or shaped blocks. Each dot may result from an individual droplet. The lines and shaped blocks may be formed from the coalescence of a plurality of droplets on the surface of the ocular prosthetic device. The array may itself form a shape, e.g. a line or shaped block, such as a ring. The dots or shaped blocks may be regular or irregular shapes, e.g. n-sided polygon, wherein n is 3 to 6, or circles, ovals, and rings. The pattern may also include one or more features selected from symbols, letters and numbers. In an embodiment, the ocular prosthetic device comprises, when viewed from above the ocular prosthetic device, a substantially circular or oval shaped body with an optic zone disposed centrally in the body, which sits on the cornea over the pupil once the ocular prosthetic device, e.g. contact lens, is fitted to an eye, and an outer zone, which sits over the cornea and sclera, The droplets, and the active ingredient and/or carrier species, may be coated on the ocular prosthetic device so that they leave a substantial portion, optionally all of, the optic zone, free of droplets, and the active ingredient and/or carrier species. The droplets, and the active ingredient and/or carrier species, may be coated on the ocular prosthetic device so that they form a ring around the periphery of the ocular prosthetic device, when viewed from above the ocular prosthetic device. The ring may be in the form of a continuous line or a discontinuous array of dots, of any shape, or lines. In an embodiment, at least 10%, optionally at least 20%, optionally at least 25% optionally at least 30%, optionally at least 35%, optionally at least 40%, optionally at least 45%, optionally at least 50% of the area of the ocular prosthetic device, on the side of the ocular prosthetic device coated, is free of droplets and active ingredient/carrier species, after the nanoelectrospraying of the ocular prosthetic device is complete, and this area is preferably in the central portion of the ocular prosthetic device (which would sit over the pupil of an eye, when in place on an eye).
After nanoelectrospraying the droplets comprising the active ingredient and/or the carrier species, any liquid from the droplets (e.g. the solvent in which the active ingredient and/or the carrier species were dispersed, e.g. dissolved) is preferably evaporated to leave just a layer comprising the active ingredient and/or the carrier species. If both a carrier species, e.g. a biocompatible polymer, and an active ingredient, e.g. a pharmaceutically active ingredient, are present on the ocular prosthetic device, this may allow a controlled release of the active ingredient to the eye once the ocular prosthetic device, e.g. contact lens, has been placed on the eye.
The thickness on the ocular prosthetic device of a layer comprising the active ingredient and/or the carrier species, after removal, e.g. evaporation, of any liquid from the droplets, is preferably 20 μm or less, preferably 15 μm or less, preferably 10 μm or less, preferably 5 μm or less. The thickness on the ocular prosthetic device of a layer comprising the active ingredient and/or the carrier species, after removal of any liquid from the droplets, is preferably 0.1 μm to 20 μm, preferably 0.5 μm to 15 μm, preferably 1 μm to 10 μm.
Optionally, multiple layers of droplets are nanoelectrosprayed onto the ocular prosthetic device. The thickness on the ocular prosthetic device of the active ingredient and/or the carrier species, after removal, e.g., evaporation, of any liquid from the droplets may vary linearly with the number of layers of droplets that are nanoelectrosprayed onto the ocular prosthetic device. In some examples, at least 10 layers are nanoelectrosprayed onto the ocular prosthetic device, for example, at least 15 layers, at least 20 layers, at least 25 layers, at least 30 layers, at least 35 layers or at least 40 layers are nanoelectrosprayed onto the ocular prosthetic device. In some examples, up to 40 layers to are nanoelectrosprayed onto the ocular prosthetic device, for example, up to 35 layers, up to 30 layers, up to 25 layers, up to 20 layers, up to 15 layers or up to 40 layers are nanoelectrosprayed onto the ocular prosthetic device. In some examples, from 10 to 40 layers are nanoelectrosprayed onto the ocular prosthetic device, for example, 15 to 35 layers, 20 to 30 layers, or 25 to 40 layers are nanoelectrosprayed onto the ocular prosthetic device. In some examples, each layer corresponds to a single rotation of the nozzle relative to the ocular prosthetic device (e.g., the contact lens) such that a layer of the active ingredient and/or the carrier species is formed on the ocular prosthetic device.
Optionally two or more active ingredients are coated onto the ocular prosthetic device. The active ingredients may perform different functions, e.g. having different therapeutic effects.
The pattern may be formed by moving the nozzle or moving the ocular prosthetic device, such that there is relative movement of the ocular prosthetic device with respect to the nozzle. The apparatus for carrying out the method may include a movable surface, on which the ocular prosthetic device resides during nanoelectrospraying. The movable surface may be movable in x and/or y directions (with the z direction being the direction from the surface to the nozzle), and x and y being perpendicular to one another and direction z. The movable surface may be movable in the x, y and z directions. Alternatively, or in addition, the nozzle may be movable in x and/or y directions (with the z direction being the direction from the surface to the nozzle), and x and y being perpendicular to one another and direction z. The nozzle may be movable in the x, y and z directions. In an embodiment, the surface on which the ocular prosthetic device reside is movable in x and y directions, while the nozzle is movable in the z direction. In an embodiment, the surface on which the ocular prosthetic device reside is movable in x and y directions, and in rotational translation (e.g. with the axis of rotation being through the centre of a circular or oval ocular prosthetic device), while the nozzle is movable in the z direction. This is depicted schematically in
In an embodiment, the relative movement of the nozzle and the prosthetic device and/or the nanoelectrospraying (e.g. control of the voltage, which may cycle between V2 and V1 as described above) is controlled by a computer (which may be any electronic device for storing and processing data according to instructions given to it in a variable program). This can allow a desired pattern, determined digitally, to be translated to a pattern on a surface of the ocular prosthetic (or other prosthetic) device, and allow this to be formed on a plurality of devices in a consistent and reproducible manner, which may lend itself to mass production of the coated ocular prosthetic devices.
In a second aspect, there is provided an ocular prosthetic device formable according to the method of the first aspect. The ocular prosthetic device may be any device for placement on or in the eye or ocular region. The ocular prosthetic device may comprise a lens. The ocular prosthetic device may be selected from a contact lens, intraocular lens, a keratoprosthesis, intraocular ring, corneal inlay (also called an intracorneal implant), and an aqueous shunts/glaucoma filtration devices.
A contact lens may be considered to be a removable prosthetic lens for the eye. The contact lens may be to correct the vision of a wearer or for cosmetic reasons or for therapeutic reasons (other than vision correction). A contact lens is typically circular or oval in shape, when looked at from above one of its largest surfaces.
The intraocular lens may comprise an optic and at least one haptic. The intraocular lens may comprise two haptics. The optic is the portion of the intraocular lens which serves as the lens and the haptic(s) is(are) attached to the optic and maintains the optic in its proper place in the eye. The optic and the haptic(s) may be of the same or different material. A multi-piece lens is so called because the optic and the haptic(s) are made separately and then the haptic(s) is(are) attached to the optic. In a single piece lens, the optic and the haptic(s) are formed out of one piece of material. In this invention, a single piece lens is preferred.
The ocular prosthetic device, onto which the droplets or active ingredient and/or carrier species are coated, may comprise any suitable material. They may comprise a material selected from polymethylmethacrylate (PMMA), poly(4-methyl-pentene), styrene-acrylonitrile copolymer, acrylonitrile-butadiene-styrene copolymer, polycarbonate, polysulfone, cellulose acetate butyrate, silicone and fluorosilicone acrylate, and a hydrogel material. A hydrogel is a polymeric material that can absorb water while maintaining its structural integrity. The hydrogel material may comprise a polymer of 2-hydroxyethyl methacrylate (PHEMA), which may be a homopolymer of HEMA or co-polymer of HEMA and one or more other monomers, with the one or more other monomers optionally being selected from N-vinyl pyrrolidone, methyl methacrylate, methacrylic acid and phosphorylcholine, a polymer of vinyl alcohol (PVA), and a copolymer of methyl methacrylate and N-vinyl pyrrolidone. The hydrogel material may be a silicone hydrogel material. The hydrogel material, on which the droplets or active ingredient and/or carrier species are coated, may comprise water in an amount of from 10% to 80% by weight of the hydrogel material, optionally from 20% to 80% by weight of the hydrogel material, optionally from 20% to 60% by weight of the hydrogel material, optionally from 30% to 50% by weight of the hydrogel material, optionally from 35% to 45% by weight of the hydrogel material.
The contact lens may be a hard contact lens or a soft contact lens.
In a third aspect, there is provided a method of coating a surface of a prosthetic device, the method comprising
nanoelectrospraying of droplets comprising an active ingredient and/or a carrier species onto the surface in a predetermined pattern, the nanoelectrospraying involving controlling the flow rate of the droplets from a nozzle of the nanoelectrospraying equipment by controlling the voltage between the nozzle and the prosthetic device. The surface may be of any curvature, e.g. curved or flat. The surface may be electrically conducting, insulating or electrically semi-conducting. Preferably, the surface is a curved and/or electrically conductive surface.
The prosthetic device may be any prosthesis for a human or animal body, i.e. an object that will act as an artificial substitute or part for a human or animal body, which may be for functional or cosmetic reasons. The prosthetic device may be selected from a stent (i.e. a prothesis for a blood vessel), a heart valve, an artificial heart or any component thereof including power supply and regulator, a breast prosthesis, a larynx prosthesis, a trachea prosthesis, a tracheobronchial prosthesis, an ear or nose prosthesis, an internal organ prosthesis, biological tissue prosthesis, a limb, hand or foot prothesis. The manner of spraying may be as described for the ocular prosthetic device. All features of the process described for the ocular prosthetic devices are equally applicable to other prosthetic devices. The active ingredient and/or carrier species may be any suitable for the prosthesis. The active ingredient may be any pharmaceutically active ingredient for treating a disorder of the human or animal body, which may include any type of active ingredient mentioned herein.
In a fourth aspect, there is provided a prosthetic device formable according to the method of the third aspect. The surface may be of any curvature, e.g. curved or flat. The surface may be electrically conducting, insulating or electrically semi-conducting. Preferably, the surface is a curved and/or electrically conductive surface. The prosthetic device may be any device with a curved and/or electrically conductive surface, such as a stent. The electrically conductive surface may be any metallic and/or wet or water-containing surface, e.g. the surface of a hydrogel material (which may be as described above for the ocular prosthetic device). The prosthetic device may be a stent having a metallic surface, onto which the active and/or carrier species has been coated using the method of the third aspect. The pattern may be any suitable pattern, e.g. an array of dots or lines or block shapes.
Standard soft contact lenses, Biomedics 38 contact lenses (CooperVision Ltd, USA), with a composition of 62% polymacon/38% H2O were used as the model lenses for coating. Ketotifen fumarate was supplied by Molekula, UK. Tetrabutylammonium bromide (TBAB), phosphate buffer saline solution tablets (pH 7.4) and poly(lactic-co-glycolic acid) 50:50 (PLGA) (brand name as Resomer RG502H) (MW7000-17000 Da) were obtained from Sigma-Aldrich, UK. Other polymers, poly(I-lactide-co-ε-caprolactone) 7:3 (PLCL, Ester terminated, Inherent viscosity: 0.93 dl/g) and poly-d,l-lactic acid (PDLLA, Ester terminated, Inherent viscosity: 0.66 dl/g) were supplied by Ashland, UK. Zein was obtained from Acros Organics, Thermo Fisher, UK), Sodium hyaluronate (MW 2 MDa) was obtained from Lifecore Biomedical. The solvents: Dichloromethane, Ethanol absolute, Acetone, Ethyl acetate were sourced from Sigma-Aldrich, UK. Milli-Q (Millipore, Merck, US) Ultra-pure water was used throughout all experiments. All materials were obtained from suppliers without further processing.
Conductivity measurements were performed by using Jenway 4510 conductivity meter with a micro volume conductivity probe. Interfacial tension of solution was measured by pendant drop method (Kyowa Interface Science, Model: DMs-401). The measurement runs for 100 second to collect the sufficient amount of data point for the analysis. For each sample, three measurements were carried out to calculate the average surface tension.
The nES process was monitored with a high-resolution camera with a high magnification zoom lens 6.5×(Thorlabs, UK). High voltage supply (F.u.G. Elektronik, Germany) was applied to the solution via a copper wire immersed in the solution. The voltage output and fast current amplifier were connected to a digital storage oscilloscope (Tektronix TBSD1000, RS component) to capture the voltage and current waveform respectively during a nES event. Different sizes of nozzles (50-100 μm) were connected to a 2 ml syringe, which was fixed on a manual 3D translation stage (as shown in
A 1% w/v stock solution of PLGA was prepared by dissolving the polymer into acetone with stirring until a clear solution was formed. Different amount of tetrabutylammonium bromide (TBAB) was added into a glass vial containing 1 ml of the PLGA stock solution to produce solutions with a range of conductivity as listed in Table 1.
Fluorine doped tin oxide glass slide was used for nES due to its conductive surface. After nES, profilometry (DektakXT Bruker) was used to measure the surface profile of the nES deposited polymer film. The Stylus radius 2 m and 1 mg Stylus force was used for the measurement. For microscopic images, Nikon LV150NL microscope with 5×-1000× magnification range was used to capture the sprayed polymer film on glass surface.
Nanoelectrospray (nES) Set Up and Operational Procedure
Nanoelectrospray printing was performed using the apparatus depicted in
An example of the applied voltage waveform for the PLGA solution to be nES printed at 1 mm NSD is shown in
As shown in
Once a suitable combination of nES process parameters of voltage, frequency, duty cycle, NSD and stage speed had been determined, the amount of deposition polymer/drug could be controlled and increased by successive overprinting onto the substrate. An example of the resulting film geometry from a single print pass and a five times overprinted substrate are detailed in
From Table 3, the effect of successive overprinting is to increase the average thickness of the printed film. Although there appears to be a small increase in width, the printed features from single pass printing and five times overprinting appear to have near identical width from optical microscopy, as shown in
Alternatives controlling nES fluid delivery rate to the substrate to achieve rapid drying are by decreasing the duty cycle and/or print frequency, increasing the stage speed or by reducing the nES-on voltage V2.
Method
Table 4 depicts the nES formulation of different polymers with or without ketotifen in organic solvents. While sodium hyaluronate (HA) was dissolved in water/ethanol (1/1) mixture and nES deposited without any drug molecule to demonstrate the spraying capabilities of high molecular weight polymer via nES technique.
nES of Drug-Polymer Solutions onto Contact Lenses
Commercial contact lenses were removed from the packaging and soaked in Milli-Q water to remove saline solution on the lens. Excess water on the lens was removed by lint-free dry wipe before being transferred onto a conductive glass substrate for nES. The drug-polymer solution was sprayed on the contact lens via nES with the parameters as shown in Table 5. A set of two lens was prepared for in vivo drug release experiment and another set was used to quantify the amount of drug being deposited on contact lens via nES.
The total amount of ketotifen deposited on a contact lens was determined by immersing the processed lens into 1 ml of the original solvent used for solubilising both the polymer and ketotifen. 0.2 ml of the solution was measured using UV absorbance at 300 nm by a quartz microplate to quantify the drug loading on the lens after nES.
Glass vials with screw caps containing 2 ml PBS pH 7.3 were first placed in a shaking incubator (IKA KS 2000i Control) set at 125 rpm and 35° C. for 2 hours to equilibrium to the environmental temperature. The contact lens was transferred immediately to the vial after the nES process for drug release study. 0.5 ml of the release sample was pipetted and replaced by same amount of PBS. At pre-determined time intervals, 0.2 ml PBS was taken and placed into a quartz microplate (Hellma, Germany). Ketotifen release was measured using UV absorbance at 300 nm using a microplate reader and analysed using the MARS Data Analysis software (ClarioStar multiwell plate reader, BMG-Labtech, UK).
The transmission of light through contact lenses was studied using a lightbox (Schott, KL1500 Electronic, MA USA), charged coupled device (CCD) camera (UVP, Cambridge, UK) and the Genesnap image capture program (Syngene, Cambridge, UK). Contact lenses were coated as described and placed onto PBS soaked gauze to maintain hydration and enclosed in a petri dish. Lenses were placed onto a petri dish base and centred over the light source. Images of control lenses (no deposition) and nES processed lenses were captured.
Images of the surface and cross section of nES lenses were taken using SEM (Zeiss, Gemini300, UK). Immediately after nES deposition, lenses were cut in half. Lenses were allowed to dehydrate and the dried lenses placed onto SEM stubs. Samples were prepared by gold sputter coating.
In order to assess the quality of the materials that is deposited on the contact lens by nES, each solution was first sprayed on a contact lens, then dissolved into 1 ml of the solvent that can completely solubilise both the model polymer and model drug on the lens. The amount of the model drug (ketotifen) deposited on the contact lens was quantified by measuring the absorbance of 200 μl of the sample at 300 nm using a quartz microplate. The average value of two lens were calculated with standard deviation (SD). As it can be seen in Table 6, the SD is very low for the model drug loaded in different polymers. This indicates that the nES material deposition is highly accurate and reproducible.
Using ketotifin fumarate as the model drug and PLCL as the model polymer, the accuracy of the nES liquid deposition around the periphery of the lens and its physical effect on the optical clarity were investigated. Using the nES parameters detailed in Table 5 to deposit a layer of the formulation containing PLCL7:3 with 10% Ketotifen Fumarate loading (PLCL/KF) creates a continuous white band after 30 rotations (
Further detailed work using SEM further demonstrates the contrast between the smooth surface of the contact lens and the layer of polymer and deposition (
The in vitro drug-release profile of model drug (ketotifen) with model polymer deposited on the contact lenses are shown in
Unless otherwise indicated, the properties mentioned herein are measured at room temperature and pressure.
In this study, nES was developed to allow precise and reproducible liquid deposition of materials on to the conductive surfaces. Using the wet contact lens as the example coating system, our data demonstrated that the nES method developed can reproducibility deposit materials in a range of patterns as dots or lines or rings. The quantity of the polymer and drug deposited on the contact lens is highly reproducible. The deposited materials allow a range of release patter of the drug to be released from the coated lenses.
Zein was obtained from Acros Organics, Thermo Fisher, UK. Ethanol absolute was sourced from VWR Chemicals, UK. Milli-Q (Millipore, Merck, US) ultra-pure water was used throughout all experiments. Phosphate buffer saline tablet and methyl blue were purchased from Sigma-Aldrich, UK. All materials were obtained from suppliers without further processing.
Preparation of Zein Solutions for Nanoelectrospraying (nES)
Zein solution was used as the example polymer solution to validate the nES process. As shown in Table 7, 2.5% w/v and 5% w/v zein solutions were prepared by dissolving zein powder in different % w/w aqueous ethanol with stirring at ambient condition until all powder dissolved. The solution was filtered through a 0.4 μm syringe filter (Fisher, UK) before nES.
Conductivity: Conductivity measurements were performed by using Jenway 4510 conductivity meter with a micro volume conductivity probe.
Viscosity: Dynamic viscosity of the solutions was measured by a Discovery HR-2 (TA instrument, Delaware, USA) equipped with a 2°, 40 mm cone-and-plate geometry. The method was set to be a flow ramp procedure from 0.1 to 100 s−1 at 25° C. for 60 seconds.
The measurement was done in triplicate to calculate the average viscosity±SD at 80 s−1.
Surface tension: Surface tension of the solutions was measured by pendant drop method (DMS-401, Kyowa interfacial science, Japan). For each sample, ten measurements were carried out to calculate the average surface tension±SD.
Density: Density of the solutions was measured by a density meter (DMA 4500M, Anton Paar, Germany) equipped with an oscillating U-tube. The measurement was done by injecting 1 ml of the sample to the system at 25° C. The measurement was done in triplicate to obtain the average density±SD.
A device constructed as shown in
There are a few spraying parameters available to be adjusted from the system, which require clarification. The motion of the 2-dimensional motorised translation stage was controlled so the substrate moved relative to the nozzle along a circular path, allowing the radius, substrate speed and number of rotations to be defined as follows. The radius indicates the relative distance between the centre of a circle at pre-set x, y coordinates to its perimeter (which corresponds to the location of the nozzle), which enables deposition of polymer film with different size rings or dots. The number of rotations controls the number of layers of spray being deposited on the surface, which enables building up of the film thickness. The substrate speed indicates the speed of the movement of the substrate in a circular motion, with a higher number indicating a faster moving speed. The nozzle substrate distance indicates the distance between the apex of the nozzle and the substrate, which is controlled by changing the coordinate of the translational stage moving in the z-axis.
A range of combinations of spraying parameters were investigated in order to understand the influence of each parameter on the width and thickness of a deposited polymer film.
These parameters are:
After nanoelectrospraying zein solutions on the conductive glass substrate, a stylus profilometer (DektakXT, Bruker, MA, USA) was used to measure the surface profile of the polymer films using the ‘Hills and Valleys’ profile. A stylus with 2 m radius was used for the measurement set at 1 mg stylus force. The measurement of spraying width and average step height (thickness) of the polymer film was performed at 3 locations as shown in
Statistical analysis was carried out using SPSS computer programme (SPSS Statistic 25). The difference in spraying width of two polymer concentrations and the viscosity difference were analysed by independent t-test (2 tailed) where a difference is considered when P<0.05 (95% confidence).
The zein solutions with two different concentrations were nanoelectrosprayed onto a conductive glass substrate to investigate the influence of polymer concentration on the morphology of the film formed. The zein solutions were sprayed with the same spraying parameters with different applied voltage (see Table 8). Profilometry results from
Different nanoelectrospraying parameters were investigated to understand their influence on the spraying width (the width of the ring-shaped film layer deposited on the substrate (a contact lens), illustrated in
For a particular formulation, the number of rotations is the main parameter that controls the thickness of the polymer film. By increasing the number of rotations, a layer by layer deposition of nanoelectrosprayed particles stack on each other to build up the thickness of polymer film. The step height (thickness) of zein film produced by the zein solution with both concentrations shows a linear relationship with the number of rotations. The results indicate that the thickness of polymer film can be controlled by the number of rotations.
For a particular formulation, the NSD is the main spraying parameter that determines the spraying width. It can be seen from
Commercial soft contact lenses, Biomedics 55 contact lenses (CooperVision Ltd, USA), with a composition of 45% ocufilcon D/55% H2O were used as the model lenses nanoelectrospraying. Ketotifen fumarate and bimatoprost were purchased from Molekula (UK). Phosphate buffer saline (PBS) solution tablets (pH 7.4), triethylamine (≥99.5%), phosphoric acid (≥85%) were obtained from Sigma-Aldrich (UK). Zein was sourced from Acros Organics, Thermo Fisher (UK). Ethanol absolute was obtained from VWR (UK). Milli-Q (Millipore, Merck, US) ultra-pure water was used to prepare all aqueous solutions. Methanol and acetonitrile, high-performance liquid chromatography grade, were purchased from Fisher Scientific (UK). All materials were obtained from suppliers without further processing. 50 μm ceramic MicroDot tip (7364054) was purchased from Nordson RFD (UK).
Nanoelectrospraying (nES) of Drug-Polymer Solutions onto Contact Lenses
Commercial contact lenses were removed from the packaging and soaked in PBS pH 7.4 for 30 minutes to equilibrate. Excess liquid on the lens was removed by lint-free wipes (RS 558-795, RS Components, UK). 20 μL of PBS 7.4 was pipetted onto the 3D printed lens holder before fitting the semi-dry contact lens on the holder (
Glass vials containing 2 ml PBS 7.4 was incubated in a shaking incubator set at 35° C. with 125 rpm for 2 hours before in vitro experiments. All in vitro experiments were performed under sink conditions (i.e., the drug concentration within the in vitro dissolution medium was more than 10 times diluted in comparison to the saturated solubility of the drug in the medium). The nanoelectrosprayed lenses were placed into the vial and an aliquot of 1.5 ml was replaced with fresh PBS at regular intervals up to 24 hours (for KF the intervals were 0.5, 1, 3, 6, and 24 hours; for BIM the intervals were 0.25, 0.5, 0.75, 1, 2, 3, 4, 5, 6, and 24 hours). Any remaining drug left on the lens was extracted by PBS until no peak was observed in an HPLC chromatogram. The total amount of ketotifen and bimatoprost sprayed on a contact lens was determined by the sum of the cumulative amount of the drug released during the in vitro study and the amount obtained from extraction.
The high-performance liquid chromatography system (Jasco, Japan) consists of a pump (PU-1580), an autosampler (AS-2055 Plus) and a 4-channel UV detector (UV-1570M). A Waters C18 column (250×4.6 mm i.d.; 5 μm particle size) was used under ambient conditions. The mobile phase of ketotifen fumarate [21] and bimatoprost [22] was prepared according to a method adopted from the literature. The mobile phase for ketotifen comprised methanol and 0.2% triethylamine (v/v) (80:20), running with 1 ml/min flow rate. The mobile phase for bimatoprost included acetonitrile, methanol and 0.1% phosphoric acid (v/v/v) (30:30:40), operating with a 1 ml/min flow rate. The injection volume was 20 μL and the detection wavelength was 300 nm and 210 nm for ketotifen and bimatoprost, respectively. Sample preparation of an aliquot was done to improve the peak intensity for both drug candidates. The aliquot collected from the in vitro experiment was mixed with the associated mobile phase in a 1:1 ratio, followed by filtration via a 0.2 μm (15141499, Fisher scientific, UK) syringe filter before the assay.
Conductivity: Conductivity measurements were performed by using a Jenway 4510 conductivity meter with a micro volume conductivity probe.
Surface tension: Surface tension of the solutions was measured by the pendant drop method (DMS-401, Kyowa interfacial science, Japan) For each sample, ten measurements were carried out to calculate the average surface tension.
Density: Density of the solutions was measured by a density meter (DMA 4500M, Anton Paar, Germany) equipped with an oscillating U-tube. The measurement was performed by injecting 1 ml of the sample into the system at 25° C. The measurement was taken in triplicate to obtain the average density.
Morphological Studies by Cryo-Scanning Electron Microscopy (cryoSEM)
Images of the surface and cross section of nanoelectrosprayed lenses were taken by using cryo-SEM (Zeiss, Gemini300, UK). Immediately after nanoelectrospraying, the lenses were cut in ⅛ of the whole lens and loaded onto an SEM stub for fast cooling with solid nitrogen. Samples were then prepared by platinum sputter coating before imaging.
Table 10 summarises the physiochemical properties of the drug-polymer formulations used for nES. The digital images shown in
According to the BNF (the British National Formulary), the strength of a ketotifen fumarate eye drop is 0.025% which is equivalent to 250 μg/mL. Taking the 5% bioavailability into consideration and the dose (1 drop twice daily), the required amount of ketotifen would be 1.25 μg per day [23]. The amount of ketotifen deposited onto a contact lens was higher than the therapeutic target in all cases. A further reduction in drug loading of ketotifen to 0.5% to obtain an even slower drug release with a sufficient dosage amount can be easily achieved to meet the therapeutic requirements.
| Number | Date | Country | Kind |
|---|---|---|---|
| 2008098.2 | May 2020 | GB | national |
There is a wide range of applications of using contact lens (CLs) as carriers to deliver and release compounds to the eye. These reported methods can be divided into 5 main categories [12, 13]: 1) drug soaking, where the lenses are typically soaked in drug solution for a period of time 2) molecular imprinting, in which a hydrogel polymer is synthesised as a complimentary template for the drug molecule to give drug binding sites within the polymer 3) layering, where typically a drug-polymer is manufactured in a layer within the lens [e.g. see U.S. Pat. No. 6,719,929 B2] 4) chemically treat and modify the lens surface [e.g. see EP 1 315 985 B1; EP 1 046 068 B1; EP 1 299 753 B1] and 5) additive coating onto the lenses by electrospinning [US 2004/0170666 A1; U.S. Pat. No. 9,956,168 B2], electrospray [US 2007/0157880 A1 EP 2 529 761 B1; EP 1988941]; or inkjet printing [EP2555751A1; US201816188833].
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/GB2021/051334 | 5/28/2021 | WO |
