DEVICE FOR TRANSMUCOSAL DELIVERY OF AGENTS

TECHNOLOGICAL FIELD

The invention generally concerns devices and methods for delivery of active and non- active agents.

BACKGROUND

Transmucosal delivery of agents is rarely used for delivering water-insoluble agents as the agents must have ideal physicochemical properties that enable their dissolution and permeation through the mucosa. Oral transmucosal (oromucosal) absorption of such agents requires their dissolution in the hydrophilic environment of the oral cavity and their permeation through the mucus membrane. Agents of high lipophilicity (Biopharmaceutics Classification System (BCS) classes II and IV) are typically poorly soluble in the oral cavity environment and additional parameters, such as high molecular weight, may have an effect on their transmucosal permeation as well. Attempts to administer highly lipophilic agents via the oromucosal route have been inefficient and resulted in poor bioavailability. Though oromucosal delivery is an attractive alternative to delivery by injection, the slow dissolution and permeation of many therapeutic agents require impractically long residence time in the oral cavity rendering oromucosal delivery less favorable.

Patent application US2011/0087192A1 [1] describes devices and methods for transmucosal drug delivery. The device may include a housing configured for intralumenal deployment (e.g., intravaginally); a drug-dispensing portion configured to dispense the drug from the housing by positive displacement; and a permeability enhancer-dispensing portion configured to release or generate a permeability enhancing substance to disrupt the mucosal barrier adjacent to the housing.

Patent application EP3065799B1 [2] provides a fluid delivery device including a housing with a porous membrane, a first fluid reservoir, a first puncture mechanism operable to puncture the first fluid reservoir, a first positive displacement mechanism operable, following puncture of the reservoir to drive the first fluid out of the first fluid reservoir. Patent application EP2900212B1 [3] provides a device for transmucosal drug delivery comprising two reservoirs comprising actuators for moving fluid (positive displacement).

PUBLICATIONS

[1] US2011/0087192A1

[2] EP3065799

[3] EP2900212

GENERAL DESCRIPTION

The inventors of the technology disclosed herein have developed devices and methods that overcome many of the difficulties associated with transmucosal delivery and thus present an effective alternative to existing inefficient methodologies.

As a person of skill in the art will appreciate, devices of the invention assure localized and tissue specific delivery (systemic delivery). Where the devices are configured for oromucosal delivery, they limit undesirable oral delivery (p.o. delivery), provide simplicity of operation and further provide precise and consistent dosing.

In a first of its aspects, the invention provides a material (e.g., agent, drug, pharmaceutical composition or formulation) dispensing device (or a mouthpiece) which comprises a plurality of channels extending to an outer surface region of the device, wherein each of the plurality of channels having a first opening and a second opening and is configured to receive and hold a liquid and permit a material flow to the outer surface of the device, wherein the material flow comprising or consisting passive flow of the liquid; or wherein the plurality of channels are formed of a plurality of hollow fibers, each having a first opening and a second opening, permitting material flow through the hollow fiber to an outer surface of the device; or wherein the plurality of channels are formed of a plurality of structured elements of at least one material, the structured elements being fused to each other to form a continuous porous material with a porosity permitting a material flow to an outer surface of the device. Also provided is a mouthpiece device having a tissue contacting surface, the device comprising a plurality of channels extending to the tissue contacting surface, wherein each of the plurality of channels having a first opening and a second opening and is configured to receive and hold a liquid material and permit a material flow to the tissue contacting surface, wherein the material flow comprising or consisting passive flow of the liquid material; or wherein the plurality of channels are formed of a plurality of hollow fibers, each having a first opening and a second opening, permitting liquid material flow through the hollow fibers to the tissue contacting surface; or wherein the plurality of channels are formed of a plurality of structured elements of at least one material, the structured elements being fused to each other to form a continuous porous material with a porosity permitting a liquid material flow to the tissue contacting surface.

In another aspect, the invention provides a material dispensing device comprising a material reservoir in liquid communication with a plurality of channels extending to an outer surface region of the device, wherein each of the plurality of channels having a first opening and a second opening, the first opening is positioned to permit a material to flow from said material reservoir through the channel to the second opening at an outer surface of the device; or wherein the plurality of channels are formed of a plurality of hollow fibers, each having a first opening and a second opening, wherein the first opening being in liquid communication with a material reservoir, permitting material flow through the hollow fiber to the second opening positioned at an outer surface of the device; or wherein the plurality of channels are formed of a plurality of structured elements of at least one material, the structured elements being fused to each other to form a continuous porous material with a porosity permitting liquid communication from the liquid reservoir to an outer surface of the device.

The invention further provides a material dispensing device comprising a material reservoir in liquid communication with a plurality of channels, e.g., microchannels, each having a first opening and a second opening, the first opening is positioned to permit a material to flow from said material reservoir through the microchannels to the second opening at an outer surface of the device. The invention further provides a material dispensing device having an outer surface comprising a plurality of openings, each of said openings being in liquid communication with another opening positioned in proximity to a material reservoir and configured to permit material flow from the material reservoir to the outer surface of the device.

Also provided is a material dispensing device, the device being formed of a plurality of hollow fibers, each having a first opening and a second opening, the first opening being at an outer surface of the device and in liquid communication with the second opening, and wherein the hollow fibers are configured to receive and hold a liquid and permit a material flow to the outer surface of the device, wherein the material flow comprising or consisting passive flow of the liquid.

Further provided is a material dispensing device, the device being in a form of a continuous porous material having a porosity permitting liquid flow to an outer surface of the device, the porosity being in a form of a plurality of microchannels, each configured to receive and hold a liquid and permit a material flow to the outer surface of the device, wherein the material flow comprising or consisting passive flow of the liquid.

Further provided is a dispensing device, the device being in a form of a continuous porous material enclosing a liquid reservoir, the continuous porous material being formed of a plurality of structured elements of the material, the structured elements being fused to each other to form the continuous porous material with a porosity permitting liquid communication from the liquid reservoir to an outer surface of the device.

In all embodiments of a device of the invention, as will be further detailed below, the outer surface of the device is configured to contact a region of a mucosal tissue to thereby enable material delivery from the device to the tissue. The so-called first openings or pores are present at the outer surface of the device. The so-called second openings are generally present at a distal end of the device, proximate to a material reservoir, in case present. Both the first and second opening are open, not clogged or stoppered, allowing passive flow of materials upon contact with a tissue, as further detailed hereinbelow. Also, in all embodiments, flow of a material from the device through microchannels or pores present in the device is in the direction of the outer surface, namely in the direction of the tissue. Back flow of materials is excluded.

In some embodiments, the structured elements are made of materials selected from particulate materials, fibrous materials, wire-type materials and fabric materials (e.g., non- woven materials). In some embodiments, fusion or otherwise association of the structured elements, as defined, forms a continuous material characterized by a porosity profile that can be controlled (a porosity profile including one or more of density of porosity, distribution of pores, size of pores and shape of pores). The porosity permits liquid communication from the liquid reservoir to an outer surface of the device.

The channels or microchannels are structures with the dispensing device through which a liquid material may be delivered from an inner section of the device to its outer surface. The channels or microchannels are typically elongated tubular structures that are “ configured to receive and hold ” an amount of the liquid material which delivery is desired. They are configured to receive the liquid by a variety of mechanisms, such as by actively injecting or forcing a liquid into the channels’ lumen, by permitting passive flow of the liquid into the lumen (e.g., when the device is soaked in the liquid and the liquid is permitted to occupy all cavities and lumens of the device) or by any other means as disclosed herein. Once substantially filled or partially filled with the liquid, the channels hold the liquid until an end of the channel is brought into contact with a tissue. The inner surface of the channel or lumen is selected of a material and optionally also selected to have an inner diameter or inner surface sufficient to achieve a surface tension which does not permit liquid escaping from the lumen. Under such conditions, liquid release is limited to passive flow mechanisms such as disclosed herein (e.g., capillary flow).

In some embodiments, the channels are microchannels.

In some embodiments, the channels inner diameter, defining the diameter of the end pores, may range from 0.01mm and 5mm.

In order to permit a material flow to an outer surface of the device, the dispensing device first needs to be filled with an amount of the material. In some embodiments, the amount of the material is filled into the device via a material reservoir, a capsule or cartridge (which are used herein interchangeably) containing the material, mostly in a liquid form.

As noted above, the material reservoir that is associated with the dispensing device may be used as a source of the material (optionally in a liquid form), enabling flow of material to an outer surface of the device optionally from the second opening. However, any other form of filling or loading the device with the material may be utilized in accordance with the present invention, such as (but not limited to) soaking the device in a liquid comprising the material to be delivered.

In some embodiments, the device may be provided completely charged with a material to be delivered. In other words, the extending channels, hollow fibers or the continuous porous material as described herein may be filled or saturated with a material, mostly in a form of a liquid, when provided for use. In some other embodiments, said channels, hollow fibers or continuous porous material are partially filled (i.e., an additional material may be added to the device before it is completely loaded).

The material to be delivered is usually in a form of a flowable liquid. The liquid may be a single material, agent or drug, a combination or such materials, or a composition, formulation, suspension or dispersion comprising at least one material, agent or drug.

Without wishing to be bound by theory or by a mechanism of action, the liquid occupying some or each the channels of a device flows in a direction of the outer surface of the device via one or more passive flow mechanisms. Such mechanisms may, without limitation, include capillary flow, gravitation, diffusion, or osmosis.

In some embodiments, the passive flow mechanism is capillary flow.

In the context of the invention presented herein, the term “ capillary flow” refers to a flow which is mostly induced by a capillary force. The term “capillary force” is to be understood as meaning the force that causes the movement of liquids in thin channels (or spaces) or through porous media, due to surface tension and/or interfacial tension acting at a liquid surface.

More specifically, capillary action occurs due to the forces of cohesion (liquid material molecules that stays close together) and adhesion (liquid material molecules are attracted and stick to other substances). Adhesion of liquid material to the walls of a hollow tube/channel/fiber will cause an upward force on the liquid at the edges. The surface tension acts to hold the surface intact. Capillary action mainly occurs when the adhesion to the walls is stronger than the cohesive forces between the liquid molecules. Eventually, a concentration gradient allows release of the material to the tissue. From the surface, liquid material molecules are absorbed into the tissue and ultimately, optionally into the bloodstream.

Since the flow of a liquid material in the channels occurs mostly via a passive flow mechanism, the device of the invention releases a first amount only when in contact with a human or animal tissue and when the tissue is capable of receiving the amount (e.g., in case the tissue is saturated or not receptive for whatever reason). Any subsequent amount discharged from the device is able to do so only when the “previous” amount was cleared from the surface tissue, namely absorbed by the tissue, thus enabling a continuous, yet gradual flow of material to the tissue, substantially without flooding the tissue or losing excessive material to the surroundings.

In aspects wherein a material reservoir exists, the reservoir may be an integral part of the dispensing device, or may be an external reservoir, which is not directly connected to the extending channels, hollow fibers or the continuous porous material.

Each of the devices of the invention, as are methods of the invention, is configured for transmucosal delivery of agents to any mucosal membrane or tissue in a human or animal body. These include mucosal tissues of the oral cavity, rectal cavity, vaginal cavity, nasal cavity as well as mucosal membranes of the eye or around the eye. The devices of the invention are not tailored nor proposed for delivery of agents to the oral cavity for achieving delivery per os.

In some embodiments, devices of the invention are configured for oromucosal delivery. In some embodiments, oromucosal delivery includes buccal and/or sublingual delivery. The delivery can be local or systemic.

In all devices of the invention, the plurality of channels or hollow fibers or structured elements, as may be the case, may be formed by any technique known in the art. Such techniques may involve (1) using a porous material, such as a porous polymer, (2) binding together a plurality of material fibers having hollow centers, (3) fusing together a plurality of material particles, fibers and/or wires which together form the two-sided open-ended channels or generally a porous structure, as defined herein, (4) 3D printing, (5) structuring a non-woven fabric product by means known in the art, enabling a functionability as described herein, or any other technique.

Irrespective of the method of production, devices of the invention may be constructed of a material which permits single or repeated use. Such materials may be selected amongst ceramic materials, polymeric materials, natural materials, fabric materials and hybrid materials. Non-limiting examples include polyethylene, polypropylene, polyesters, polyamides, cellulose based materials, natural fibers and others.

Where the device is constructed of fused hollow fibers, each of the hollow fiber, having a tubular shape, may be selected based on its length, inner diameter, wall thickness, internal surface roughness and the material from which it is made. A device of the invention may be made of fused fibers which are substantially the same or different in one or more of length, inner/external diameter, wall thickness and material. In fact, irrespective of the means of preparation, devices of the invention having a plurality of channels, having each two or more openings, may be selected, designed, configured, assembled and/or engineered to meet one or more criteria relating to the agent to be delivered, the agent’s physiochemical properties, the agent’s particle size, the agent’s hydrophobic properties/hydrophilic properties, the receptor mucosal tissue, the delivery duration required for the complete administration of the agent, the amount of agent to be delivered, a desired volume or quantity of a droplet or a quanta amount of the material (solution, dispersion or emulsion) to be delivered, the polarity of the material, the material physiochemical properties and other parameters. Such criteria may be met by selecting one or more of the channels’ length, inner or external diameter, surface roughness, wall thickness and material.

The pores formed at the outer surface of the device, defining channel ends, do not extend outwards of the device outer surface and are not configured to insert into the mucosal membrane. The devices of the invention do not utilize microneedles, permeability enhancers or any alternative means to puncture, injure or penetrate into the membrane.

The at least one liquid material may comprise (consist) an agent in its pure form (a liquid material), or an agent carried in a liquid carrier forming a solution, a dispersion or an emulsion. The agent may be any such material which dispensing onto a living mucosal tissue membrane is desired. Typically, the agent may be selected amongst pharmaceutical agents, nutraceuticals, dietary supplements, vitamins and minerals and others, as may be the case. Such agents may be water soluble or water insoluble or characterized by low water solubility. In some cases, the agent is a lipophilic agent or an agent that is hydrophobic or non-hydrophilic.

In some embodiments, the agent is characterized by a high LogP value.

When the agent is a pharmaceutical material or a biopharmaceutical material it may be categorized as acceptable in the field, e.g., as a Biopharmaceutics Classification System (BCS) class II or IV.

Without wishing to be bound by theory or a particular mode of operation, the at least one liquid material flows from the reservoir (when such exists) and/or through the channels or pores. Typically, the flow proceeds from the ends of the channel end close to or in proximity to or in contact with or which is immersed in the material reservoir (namely, the first opening) to the other end of the channel, positioned at the outer surface of the device (namely, the second opening) which is configured to intimately contact the mucosal membrane. Droplets of the at least one liquid material advance continuously through the channels, either by an active (on demand production of droplets) or by a passive transport mechanism. Where active flow is desired, on-demand production of droplets may be initiated using valves or electric actuators, as may be the case. Where passive flow such as a capillary flow, gravitation, diffusion or osmosis is desired, continuous droplet production may be achieved by proper construction of the channels geometry, e.g., volume, lumen length, position etc.

Pressure-driven flow may also be utilized.

Devices of the invention permit utilization and absorption of any type of a liquid material or a solution whether lipophilic or hydrophilic. This is feasible by dint of the passive flow mechanism which naturally “holds” the flow until after a previous amount of the material is absorbed into the selected tissues. Thus, no substantial loss or spillage of the material is observed. Materials having lower absorbance rates will flow in a slower manner to the outer surface of the device due to the artificial barrier of liquid material which is formed between the absorbing tissue and the device. Such a barrier is a bottleneck which slows the flow at least at the contact area between the device and the desired tissue. When external forces are applied to drive the liquid material out of the lumen as described, the flow rate may increase.

Droplets containing the at least one liquid material are produced when the material, e.g., in a hydrophobic or non -hydrophilic form, contacts the mucosal membrane and the aqueous environment surrounding it. The dosage or amount of the agent that comes into contact with the mucosal membrane and which eventually becomes absorbed through the tissue depends on, inter alia, the shape and size of the dispensing device, the area of contact, the porosity profile of the device outer surface, namely on the number of channels, structured elements or hollow fibers that extend the material reservoir and the outer surface per unit surface area, the average pore size and the material from which the device is made, just to name a few. Both the number of pores and their size (diameter) define the volume of the material that exists the device (calculated based on the number of pores and the volume of each of the droplets exiting) and the rate at which the material volume is discharged from the ends of the channels. By selecting a specific porosity profile, size/geometry and a device material (or at least the material of the inner channels surface) one can control the volume of the material that comes into contact or remains in contact with the tissue before the next volume is discharged, thereby rendering efficient the tissue absorbance and avoiding material flooding the mucosal tissue. By selecting a porosity profile, size/geometry and a device material, one may further control discharge of the at least one liquid material from the device pores. In fact, the porosity profile, size, geometry and device material are selected to prevent or arrest discharge of the at least one liquid material when the device is not in intimate contact with the mucosal membrane. These may further enable to minimize the loss of the agent, the control in the flow rate, increase the control and consistency that should increase efficiency of the dissolution and the permeability of the agent, reduce the chances of staining due to the “still” mode and increase the dispersal control in the delivered area and/or deduction of the intra and inter user variance.

The total volume (dosage) of the material is typically predetermined. In using a device of the invention, the amount of the material (in volume units) may be predetermined prior to use. In such cases, the amount of the material may be sufficient for a complete treatment cycle. In some cases, the device is configured for a single use or multiple uses. The amount of the agent to be delivered may range from micrograms to hundreds of milligrams or even grams. The amount may depend on a variety of factors ranging from allowable amounts or prescribed amounts, and may depend on additional factors such as the agent, the concentration of the agent to be delivered, the carrier liquid, the surface area of the mucosal membrane, the capacity of the membrane to absorb the agent, the rate of discharge from the device, which may depend on the device porosity profile, as explained herein, on the duration of device operation and on other factors.

In some embodiments, the amount of the agent may range between 0.001 and 500 mg, in a volume ranging between 0.5 milliliter to 100 milliliter.

As stated herein, the at least one liquid material may be presented in pure form or in a liquid carrier (medium), in a form of a solution, a suspension or a dispersion. The liquid carrier may be selected amongst water, water containing media, oils, alcohols, and other pharmaceutically acceptable carriers. The liquid material may include additional excipients, acting as adjuvants, solubility enhancers, absorption enhancers, stabilizers, preservatives and the like, commonly used for pharmaceutical formulation. These carriers may be used as vehicles, adjuvants, excipients or diluents and are well- known to those skilled in the art and are readily available to the public.

Oils which can be used in devices of the invention as media for dissolving, suspension or dispersing the material may be selected from petroleum, animal, botanical, vegetable, or synthetic oils. Specific examples of oils include peanut, soybean, sesame, cottonseed, corn, olive, coconut, petrolatum and mineral oils. Suitable fatty acids include oleic acid, stearic acid and iso-stearic acid. Ethyl oleate and isopropyl myristate may also be used.

The agent to be delivered or administered to the mucosal membrane may be any such material which dispensing onto a mucosal tissue is desired and may be selected as defined hereinabove. In some embodiments, the agent is selected from pharmaceutically active agents, drugs, additives, supplements and others. Non-limiting examples of such materials include vitamins, proteins, anti-oxidants, peptides, polypeptides, lipids, carbohydrates, hormones, antibodies, monoclonal antibodies, therapeutic agents, antibiotic agents, vaccines, prophylactic agents, diagnostic agents, contrasting agents, nucleic acids, nutraceutical agents, small molecules of a molecular weight of less than about 1,000 Da or less than about 500 Da, electrolytes, drugs, immunological agents, macromolecules, biomacromolecules, analgesics or anti-inflammatory agents; enthelmintic agents; anti-arrhythmic agents; anti-bacterial agents; anti-coagulants; anti depressants; antidiabetics; anti-epileptics; anti-fungal agents; anti-gout agents; anti hypertensive agents; anti-malarial agents; anti-migraine agents; anti-muscarinic agents; anti-neuroplastic agents or immunosuppressants; anti-protazoal agents; anti-thyroid agents; alixiolytics, sedatives, hypnotic or neuroleptic agents; beta-blockers; cardiac inotropic agents; corticosteroids; diuretic agents; anti-Parkinsonian agents; gastro intestinal agents; histamine Hl-receptor antagonists; lipid regulating agents; nitrate or anti-anginal agents; nutritional agents; HIV protease inhibitors; opioid analgesics; cytotoxic agents; cannabinoids; terpenoids, flavonoids, rescue drugs; and stimulants.

In some embodiments, the active agent is at least one cannabinoid, optionally in combination with one or more other material derived from the cannabis plant, e.g., flavonoids and terpenoids.

As known in the art, the cannabinoids encompass a class of chemical compounds, cannabinoid/cannabinoid agonists/cannabinoid-related compounds, acting with various affinities on the endogenous cannabinoid receptors (CB1 and CB2). In accordance with aspects and embodiments of the invention, the term encompasses the group of ligands that include the endocannabinoids (produced naturally by humans and animals), phytocannabinoids (found in cannabis and some other plants), and synthetic cannabinoids (manufactured artificially). The most notable are tetrahydrocannabinol (THC) and cannabidiol (CBD).

In some embodiments, the material to be administered by utilizing devices as herein is at least one of a Tetrahydrocannabinol and Cannabinol-type (THC, CBN), Cannabidiol-type (CBD), Cannabigerol-type (CBG), Cannabichromene-type (CBC), Cannabielsoin-type (CBE), iso-Tetrahydrocannabinol-type (iso-THC), Cannabicyclol- type (CBL), Cannabicitran-type (CBT), a derivative, a precursor, or a combination thereof. All classes derived from cannabigerol-type compounds and differ mainly in the way this precursor is cyclized. The classical cannabinoids are derived from their respective 2-carboxylic acids (2-COOH, also denoted with -A) by decarboxylation (catalyzed by heat, light, or alkaline conditions). In some embodiments, the active agent is tetrahydrocannabinol or cannabidiol acid precursors, THC-A or CBD-A.

In some embodiments, the cannabinoid is selected from tetrahydrocannabinol (THC), cannabidiol (CBD) and cannabinol (CBN).

In some embodiments, the at least cannabinoid is an extract derived from a cannabis plant and which comprises one or more of the aforementioned cannabinoid materials.

In some embodiments, the active agent is provided in a water-soluble form, e.g., in an ionic form, a complex, encapsulated, or in any other form.

In some embodiments, the active agent is provided in the form of an active combination of at least one active agent and at least one other active or non-active agent.

In further embodiments, the active agent may be provided in the form of an active formulation comprising at least one active or non-active agent and one or more materials.

In some embodiments, the dispensing device of the invention is provided as part of a delivery apparatus which comprises the dispensing device, an electronic unit and a user interface. In other words, a delivery apparatus is any apparatus comprising at least the dispensing device as provided in the herein disclosure and any other elements which may be essential or required to operate, control, or load the dispensing device.

The dispensing device may be formed as a mouthpiece and structured based on the particular oromucosal administration desired. For example, for buccal delivery, the device may be flat or rounded to enable intimate contact with the buccal tissue and convenience to the user. For sublingual delivery, the device may be shaped in a U shape to intimately fit underneath the tongue. The device may allow 360°, two-sided delivery or a one-sided delivery of the material.

In the context of the herein disclosure, the terms “dispensing device”, “mouthpiece” and “applicator” are interchangeable and refer to the part of the device containing the plurality of channels/hollow fibers/porous material, as disclosed herein. The material reservoir may or may not be an integral portion of the dispensing device, mouthpiece or applicator, all in accordance with the specific embodiment, where the terms appear. In some embodiments, an oromucosal device may be configured for delivery to any other mucosal membrane in a subject’s body, including rectal, nasal, ocular and vaginal delivery and thus may be shaped to fit such application.

Notwithstanding the shape and size of the device, the material reservoir (when needed) may be shaped and positioned to achieve an effective liquid communication with the outer surface of the device through the plurality of channels, structured elements or hollow fibers. Generally speaking, the material from which the device is constructed, as detailed herein, may be selected to arrest diffusion or permeation of the active or non-active material or a medium in which it is contained through the bulk of the device. Thus, the material reservoir may be contained or enveloped in a material that on one hand permits liquid communication with the channels’ openings but at the same time limits contact with the device bulk material and minimize material loss, i.e. agent.

In some embodiments, the reservoir is in the form of one or more capsules that is/are (each) loaded with a single dose or multiple doses of the material and is/are disposed after treatment or is/are reusable (non-disposable). Such a capsule may be configured to liquid communicate with the plurality of channels.

The invention also contemplates a method for delivering at least one agent to a mucosal membrane of a subject, the method comprising contacting a region of the mucosal membrane with a dispensing device or apparatus comprising the at least one agent; the device being configured to permit metered delivery of the at least one agent to the membrane such that each metered amount is dispensed from the device when the device is in contact with the membrane and after a prior metered amount has been absorbed by the membrane.

In some embodiments, the metered delivery is via passive flow mechanisms. In some specific embodiments, the passive flow mechanism is a capillary flow.

In some embodiments, the dispensing device is provided with a membrane contacting surface having a porosity enabling dispensing the at least one agent.

In some embodiments, the dispensing device is provided with a membrane contacting surface that comprises pores that are in liquid communication with a material reservoir comprising the at least one agent. In some embodiments, the pores defining the ends of a plurality of (micro)channels extending from the contacting surface to the material reservoir (if one exists), thereby permitting liquid communication.

In embodiments where the device is filled with a liquid (e.g., being soaked in a liquid), the material reservoir is not necessary, since such a device can operate both as a reservoir for holding the liquid (particularly the channels) and as a dispensing unit, in a way that liquid material flows within the lumen of said channels/hollow fibers/porous material to the outer surface of the device.

In some embodiments, the dispensing device is the device described hereinabove.

Further provided is a kit or a commercial package comprising the dispensing device or apparatus as described herein.

In some embodiments, the kit or commercial package comprises a plurality of devices or apparatus as described herein.

In some embodiments, the kit or commercial package comprises a single device or apparatus and a plurality of disposable reservoirs, optionally in a form of a cartridge, each comprising an amount of a material, mostly in a liquid form. The reservoirs are configured to provide material to the device which passively flows within the channels/fibers/porous material as described to the outer surface of the device.

In some embodiments, the kit further comprising a remote controller, or a mobile application configured to control or monitor efficient or complete delivery of the agent, or to control or monitor proper operation of the device. Such remote controller or application may control the amount, volume or rate of release, or any other property of the device.

In some embodiments, the kit comprises instructions of use.

Thus, aspects and emboidemnts of the invention relate to any one of more of the following:

A dispensing device comprising a material reservoir in liquid communication with a plurality of channels extending to an outer surface region of the device, wherein each of the plurality of channels having a first opening and a second opening, wherein the first opening is positioned to permit a material to flow from a material reservoir to the second opening at an outer surface of the device; or wherein the plurality of channels are formed of a plurality of hollow fibers, each having a first opening and a second opening, wherein the first opening being in liquid communication with a material reservoir, permitting material flow through the hollow fiber to the second opening positioned at an outer surface of the device; or wherein the plurality of channels are formed of a plurality of structured hollow elements of at least one material, the structured hollow elements being fused to each other to form a continuous porous material with a porosity permitting liquid communication from a liquid reservoir to an outer surface of the device.

In a device of the invention, the material reservoir may be detachable from the device.

A device of the invention comprises a material reservoir in liquid communication with a plurality of channels, each of said plurality of channels having a first opening and a second opening and a lumen extending the first and second openings, wherein the first opening is positioned to permit a material to flow from said material reservoir through the lumen to the second opening at an outer surface of the device.

A device of the invention has an outer surface comprising a plurality of openings, each of said openings being in liquid communication with another opening positioned in proximity to a material reservoir to permit material flow from the material reservoir to the outer surface of the device.

A device of the invention is formed of a plurality of hollow fibers, each of said plurality of hollow fibers having a first opening and a second opening, the first opening being in liquid communication with a material reservoir, permitting material flow through the hollow fiber to the second opening positioned at an outer surface of the device.

The device of the invention is alternatively formed of a continuous porous material enclosing a liquid reservoir and formed of a plurality of particles of the material, wherein the material having a porosity permitting liquid communication from the liquid reservoir to an outer surface of the device.

The device of the invention is configured for transmucosal delivery of at least one agent to a mucosal membrane or tissue in a human or animal body, the mucosal membrane being a mucosal tissue of the oral cavity, rectal cavity, vaginal cavity, nasal cavity or mucosal membranes of the eye.

The device of the invention is configured for oromucosal delivery.

The device of the invention is configured for oromucosal delivery, e.g., buccal or sublingual delivery.

The device of the invention is configured for single use or multiple uses.

The device of the invention is formed of a material selected from ceramic materials, polymeric materials, natural materials, fabric materials and hybrid materials.

The device of the invention is of a material that is or comprises polyethylene, polypropylene, polyesters, polyamides, cellulose based materials and/or natural fibers.

In a device of the invention, each of the channels having an inner diameter ranging from 0.1mm to 5mm.

In a device of the invention, the liquid reservoir comprises at least one agent selected from active and non-active agents.

In a device of the invention, the at least one agent is provided in neat form, or carried in a liquid carrier.

In a device of the invention, the at least one agent is provided in a solution, a dispersion or an emulsion.

In a device of the invention, the at least one agent is selected amongst pharmaceutical agents, nutraceuticals, dietary supplements, vitamins and minerals.

In a device of the invention, the at least one agent is water soluble or water insoluble or characterized by low water solubility.

In a device of the invention, the at least one agent is a lipophilic agent or a non- hydrophilic agent.

In a device of the invention, the at least one agent is a pharmaceutical material or a biopharmaceutical material categorized as a Biopharmaceutics Classification System (BCS) class II or IV.

In a device of the invention, the liquid carrier is selected amongst water, water containing media, oils, alcohols, and pharmaceutically acceptable carriers.

In a device of the invention, the at least one agent is selected from pharmaceutically active agents, drugs, additives, supplements and others. In a device of the invention, the at least one agent is selected from vitamins, proteins, anti-oxidants, peptides, polypeptides, lipids, carbohydrates, hormones, antibodies, monoclonal antibodies, therapeutic agents, antibiotic agents, vaccines, prophylactic agents, diagnostic agents, contrasting agents, nucleic acids, nutraceutical agents, small molecules of a molecular weight of less than about 1,000 Da or less than about 500 Da, electrolytes, drugs, immunological agents, macromolecules, biomacromolecules, analgesics or anti-inflammatory agents; enthelmintic agents; anti- arrhythmic agents; anti-bacterial agents; anti-coagulants; anti-depressants; antidiabetics; anti-epileptics; anti-fungal agents; anti-gout agents; anti-hypertensive agents; anti- malarial agents; anti-migraine agents; anti-muscarinic agents; anti-neuroplastic agents or immunosuppressants; anti-protazoal agents; anti-thyroid agents; alixiolytics, sedatives, hypnotic or neuroleptic agents; beta-blockers; cardiac inotropic agents; corticosteroids; diuretic agents; anti-Parkinsonian agents; gastro-intestinal agents; histamine Hl-receptor antagonists; lipid regulating agents; nitrate or anti-anginal agents; nutritional agents; HIV protease inhibitors; opioid analgesics; cytotoxic agents; cannabinoids; terpenoids, flavonoids, rescue drugs; and stimulants.

In a device of the invention, the at least one agent is at least one cannabinoid, optionally in combination with one or more other material derived from the cannabis plant.

In a device of the invention, the at least one cannabinoid is a cannabis extract.

In a device of the invention, the at least one agent is provided in a water-soluble form.

In a device of the invention, the water-soluble form is selected from an ionic form, a complex and an encapsulated form.

A device of the invention is for dispensing an amount of the at least one agent or a formulation or a dispersion or an emulsion comprising same, the amount being determined by one or more of surface area of the mucosal membrane, capacity of the membrane to absorb the at least one agent, rate of discharge from the device and duration of device operation.

A device of the invention is for dispensing an amount between 0.001 and 500 mg, when in a volume ranging between 0.5 milliliter to 100 milliliter. A device of the invention is configured as a mouthpiece.

A device of the invention is configured for sublingual delivery of the at least one agent.

A device of the invention is provided in a delivery apparatus further comprising an electronic unit and a user interface.

A delivery apparatus is provided which comprises a dispensing device according to the invention.

A method is provided for delivering at least one agent to a mucosal membrane of a subject, the method comprising contacting a region of the mucosal membrane with a dispensing device comprising the at least one agent; the device being configured to permit passive metered delivery of the at least one agent to the membrane such that each metered amount is dispensed from the device when the device is in contact with the membrane and after a prior metered amount has been absorbed by the membrane.

In a method of the invention, the dispensing device is provided with a membrane contacting surface having a porosity enabling dispensing the at least one agent.

In a method of the invention, the dispensing device is provided with a membrane contacting surface that comprises pores that are in liquid communication with a material reservoir comprising the at least one agent.

In a method of the invention, the dispensing device is the device s disclosed herein.

A mouthpiece device is provided having a tissue contacting surface, the device comprising a plurality of channels extending to the tissue contacting surface, wherein each of the plurality of channels having a first opening and a second opening and is configured to receive and hold a liquid material and permit a material flow to the tissue contacting surface, wherein the material flow comprising or consisting passive flow of the liquid material; or wherein the plurality of channels are formed of a plurality of hollow fibers, each having a first opening and a second opening, permitting liquid material flow through the hollow fibers to the tissue contacting surface; or wherein the plurality of channels are formed of a plurality of structured elements of at least one material, the structured elements being fused to each other to form a continuous porous material with a porosity permitting a liquid material flow to the tissue contacting surface.

BRIEF DESCRIPTION OF THE DRAWINGS

In order to better understand the subject matter that is disclosed herein and to exemplify how it may be carried out in practice, embodiments will now be described, by way of non-limiting example only, with reference to the accompanying drawings, in which:

Fig. 1 depicts an exemplary delivery apparatus according to the invention.

Figs. 2A-C depict different dispensing devices, e.g., mouthpieces, according to certain embodiments of the invention.

Fig. 3 depicts another exemplary delivery apparatus according to the invention.

Fig. 4 depicts another exemplary S -shaped delivery apparatus including a thumb handle for conveniently holding the device in its place.

Fig. 5 provides a front view and a side view of yet another exemplary delivery apparatus of the invention.

Fig. 6 depicts an exemplary delivery apparatus; the apparatus includes a cartridge/capsule/material, a capsule chamber for placing the cartridge/capsule/material reservoir inside, an indicator on the capsule chamber of the administered amount or the remaining amount of liquid material, an applicator (a dispensing device of the herein invention) comprising said plurality of extending channels/fibers/porous material, a rotation mechanism which, upon rotation, enables a liquid communication between the reservoir and the applicator, an upper lid enclosing and securing the capsule in its place and a front lid enclosing and protecting the applicator.

Fig. 7 depicts the area to which the device/apparatus is placed to permit efficient oromucosal material delivery. In the figure, the device/apparatus is placed sublingually.

Fig. 8 shows rat cadaver #829 on an experiment to observe the absorption of colorants into the buccal/sublingual tissues via the applicator of the invention at t=0 (start of experiment). Oil soluble green colorant is used.

Fig. 9 shows rat cadaver #829 after 60 minutes with the applicator under the tongue. Figs. 10A-B show rat cadaver #829 after 60 minutes with the applicator under the tongue, after removal of the applicator, before wiping the excess of oil.

Fig. 11 shows rat cadaver #829 after 60 minutes with the applicator under the tongue, after removal of the applicator, after wiping the excess of oil. A green color is spotted in the circled area.

Fig. 12 shows rat cadaver #831 on an experiment to observe the absorption of colorants into the buccal tissues via the applicator of the invention at t=0 (start of experiment). Water soluble trypan blue colorant is used.

Fig. 13 shows rat cadaver #831 after 60 minutes with the applicator under the tongue, after removal of the applicator, before wiping the excess of the blue colorant.

Fig. 14A-B show rat cadaver #831 after 60 minutes with the applicator under the tongue, after removal of the applicator, after wiping the excess of blue colorant. Blue color is spotted in the circled areas.

Fig. 15 shows the mean concentration (ng/ml) of midazolam in blood samples of the rats of group #1 (rats 1-4) over time.

Fig. 16 shows the mean concentration (ng/ml) of midazolam in blood samples of the rats of group #2 (rats 5-8) over time.

DETAILED DESCRIPTION OF EMBODIMENTS OF THE INVENTION

EXAMPLE 1 - DEVICE AND METHOD

An exemplary delivery apparatus shaped as a pen device is shown in Fig. 1.

The dispensing device is configured in this example for oromucosal delivery with an exemplary disposable mouthpiece (dispensing device) shown in Fig. 2A. An alternative mouthpiece is shown in Fig. 2B.

In the pen of Fig. 1, the mouthpiece is configured to removably associate or fit into the body of the delivery apparatus which may be configured to allow mechanical or electronic operation. In the shown example, an electronic unit operably allows communication of a liquid material from a reservoir, e.g., a disposable capsule or a cartridge (however, generally the device does not necessarily require a reservoir to be fully operable as described above). The ON/OFF operation of the apparatus may be controlled from the electronic unit which can further provide a dial or a general means to set the dose or volume to be delivered, the duration of delivery and other parameters relating to the dose or volume. The operation of the device may be local via pressing a button or by distant control from a distant device such as mobile device via Bluetooth or any other technology which permits distant communication.

To control or monitor efficient or complete delivery of the agent, or to control or monitor proper operation of the apparatus, the apparatus may further comprise a timer for timing the beginning and/or end or duration of the delivery; a sensor for controlling or monitoring rate of delivery or flow of the agent from the reservoir or any other parameter of operation; and others.

Operation of the apparatus may be set by a timer, or by setting the ON/OFF switch or dial to ON or communicating with the apparatus when use is intended. Dispensing of the material may commence however only upon contacting the mouthpiece with the mucosal membrane. In some configurations, an ON/OFF switch or dial may not be required.

Particularly where the apparatus does not comprise or does not utilize electronic elements, the apparatus may comprise the dispensing device, shaped as described herein, comprising a dispensing unit comprising a plurality of channels/hollow fibers/continuous porous material extending to an outer surface region of the device, wherein each of the plurality of channels having a first opening and a second opening and are configured to receive and hold a liquid and permit a material flow to the outer surface of the device, wherein the material flow comprising or consisting passive flow of the liquid and optionally one or more reservoirs (or cartridges or capsules) that are in liquid communication with the unit. Delivery of the agents from the reservoir(s) may ensue by mechanically setting the device to ON and contacting the mucosal membrane therewith.

In some configurations, the device may comprise a reservoir which is not directly linked to the dispensing unit (i.e., an external reservoir), however such a reservoir is able to be in a liquid communication with the dispensing unit when needed.

In some configurations, the reservoir is directly connected to the dispensing unit and thereby allows flow of liquid material therein.

In some embodiments, the flow is by passive flow mechanisms.

In some configurations the device may comprise a dispensing unit per se which is prefilled with a material to be delivered to the selected tissues. In some embodiments, the prefilling is obtained by soaking the device in a desired liquid material.

In some configurations of an apparatus of the invention, an ON/OFF switch, either mechanical or electronic, is not required as dispensing of the active materials can occur only when the dispensing unit comes into contact with the mucosal membrane.

Not wishing to be bound by any one embodiment of an apparatus, or dispensing device or method of the invention, as disclosed herein, the apparatus shown in Fig. 1 need not comprise a dose calibrator. Dispensing of the dose may not require calibration or control. For example, the apparatus may be configured for continuous and complete delivery of a volume of a reservoir or a capsule. In such cases, a dose calibrator may not be required. In other cases, the reservoir or capsule may comprise a material volume equivalent of two or more doses and thus sufficient for multiple uses. In such cases a dose or a volume calibrator operable to arrest dispensing after a predetermined volume is dispensed may be included.

In some other embodiments, the apparatus of Fig. 1 or any other apparatus provided by the herein invention need not comprise a reservoir. In such embodiments, the device is pre filled with a material to be delivered (optionally by soaking).

Where an apparatus of the invention is electronically operated, the apparatus may comprise a power supply, e.g., a battery, a rechargeable battery, an electronic charger, means for connecting to a power supply, etc.

The operation of the apparatus or system of the invention may require the use of a sensor or an array of sensors for monitoring proper operation and for signaling, e.g., initiation and completion of treatment cycle(s).

A user interface panel may be provided to permit operation of the apparatus and selection of parameters such as dosage size.

The dispensing devices/units examples shown in Figs. 2A-C are shaped as mouthpieces (Figs. 2A and 2B), e.g., for sublingual delivery, or for inner cavity (Fig. 2C: rectal, vaginal, nasal- at different dimensions). In Fig. 2A, the black dots designate the distribution of channel openings extending from the core of the device to its outer surface. The density and pore diameter defining the device porosity profile. Similar pore structure may also be applicable to other configurations of the device, such as those exemplified in Figs. 2B and 2C. The invention further provides a method for delivering at least one agent to a mucosal membrane of a subject (human or non-human), the method comprising:

-contacting a region of the mucosal membrane with a dispensing device comprising the at least one agent; the device being configured to permit metered (e.g., dropwise) delivery of the agent to the tissue such that each metered amount (e.g., each drop) is dispensed from the device when the device is in contact with the tissue and after a prior metered amount has been absorbed by the tissue.

Methods of the invention are further directed to using any one device as disclosed herein.

In some embodiments, the dispensing device is provided with a membrane contacting surface (the device outermost surface) having a porosity enabling dispensing the at least one agent. The porosity, e.g., the number of pores per unit surface area and the size of the pores, determines the metered amount to be dispensed.

In some embodiments, the dispensing device is provided with a membrane contacting surface that comprises pores that are in liquid communication with a material reservoir comprising the at least one agent.

In some embodiments, the pores defining the ends of a plurality of (micro)channels extending from the contacting surface to the material reservoir, thereby permitting liquid communication.

Devices/apparatus of the invention are designed to be used in at least two configurations:

- According to a first configuration, the device (as provided in Figs. 3-6) is placed in the sublingual/buccal area (as depicted in Fig. 7) and held in such position by a subject or caregiver. The device is held in position until the treatment is over, and the full amount intended to be administered is given to the subject. The indicator (Fig. 6) on the cartridge displays the given amount. In such a configuration, the subject or the caregiver can set the dose or volume to be delivered, the duration and rate of delivery and other parameters relating to the dose, volume and/or rate. The operation of the device may be local via pressing the buttons or by distant control from a distant device such as mobile device via Bluetooth or any other technology which permits distant communication. The duration of administration, rate of delivery or flow of the agent from the reservoir or any other parameter of operation is controllable. According to a second configuration, the dispensing unit (mouthpiece/applicator) alone is retained in the sublingual/buccal area. In some examples, the mouthpiece is prefilled by the operation of the apparatus (with the material comprised in the material reservoir/cartridge). In other examples, the mouthpiece is soaked and thereby loaded with a desired amount of material to be delivered and is placed in the sublingual/buccal area of a subject.

In any of the above configurations, the dose or volume to be delivered, the duration of delivery and other parameters relating to the dose, volume or rate are fully controllable as described above.

EXAMPLE 2 - PHARMACOKINETICS

Provided herein is an evaluation of safety and PK parameters of liquid medicine in sublingual administration via devices and methods of the invention. Further described is the onset time, C _max , T _niax. AUC and relative bioavailability of sublingual administration with and without the device.

Also provided is examination of the timing of bleeding points and the initial comparison to traditional sublingual administration.

Materials

Applicator / Device

Table 1: describes device’s specifications.

Active liquid loaded into the device

In the present example, midazolam (Buccolam© - pre-filled oral syringe of oromucosal solution comprising 10 mg midazolam (as hydrochloride) in 2 ml solution - 5mg in 1ml) for treating prolonged, acute, convulsive seizures in infants, toddlers, children and adolescents (from 3 months to < 18 years) has been loaded into the device. The dose should be administered in accordance with the age as presented in Table 2 below. Pharmacokinetic properties of Buccolam© are presented in Table 3. Posology (Buccolam©)

Table 2: shows the corresponding dose to the age range for treating with Buccolam©.

Pharmacokinetics (Buccolam©)

Table 3: shows the pharmacokinetic properties and the corresponding dose and age range for Buccolam©.

Elimination

Plasma clearance of midazolam in children following oromucosal administration is 30 ml/kg/min. The initial and terminal elimination half-lives are 27 and 204 minutes, respectively. Midazolam is excreted mainly by renal route (60-80% of the injected dose). Less than 1% of the dose is recovered in urine as unchanged active (i.e., most of the active is metabolised via hepatic CYP450 enzymes, and particularly CYP450-3A4).

Method Animals

8 Adult male healthy rats (weights 250 g each) were used in the study.

Procedure

Treatment groups: 8 rats (n=8) divided into 2 groups.

Group #1 includes 4 rats which received the active via the novel device; Group #2 includes 4 rats which received the active sublingually in a direct manner via pipette or syringe.

The active was administered via the device by sublingual route (under the tongue and the floor of the mouth). The active has also been administered via traditional SL (sublingual) administration by dripping Buccolam© solution with a pipette/syringe. Study groups as demonstrated below are tested on the same day and in some cases over 2 consecutive days.

In the experiments, the actual dosage given to the animals may vary from the above calculations, without compromising study results (e.g. due to residency time). Thus, to control such possible variation and allow comparison between study groups, the dose given to Group #2 (traditional SL administration) will be the calculated mean dose given to Group #1 via the device.

Blood samples are 125±10pL of plasma (0.25mL full blood) each.

The dispensing unit (applicator) is being weighted after 30 minutes (where t=0 is the time where the applicator is properly sublingually placed and is in contact with mouth tissues). Table 4 below, provides the division into 2 groups, each group is unique by the mode of administration. In the table, sampling time points in minutes are also provided.

Table 4: sampling time points in minutes for the 2 different study groups

Sequence of operation

Group #1- device

1. Empty half of Buccolam© 10mg/2ml syringe into a small beaker 2. For each animal: a. Pick a single device (as described above) b. Weigh the dry device and record the result (nominal weight = 0.337mg) c. Soak the applicator in Buccolam© formulation for 20 seconds until it is fully soaked d. Gently remove residual drops from the device surface e. Weigh the fully soaked applicator and record the result in the table below (nominal weight = 0.537). i. If delta is <200mg (between the wet and the dry applicators) re soak and record the result. f. Place the applicator in the sublingual area with the flat wide end proximally positioned in a way which permits i. maximal surface contact with the back of the tongue and the floor of the mouth g. Start the timer for each rat h. Take photos for record i. Take blood samples at time points as described in Fig. 20 (mean concentrations of midazolam in said samples are recorded in the figure). j. At t=30 min i. Remove the device with a pair of tweezers ii. Weigh the device and record the weight in a table

Group #2- Syringe/pipette

1. Calculate the mean dose of Group 1 and set it as the starting Group 2 dose.

2. Use the residual Buccolam© from Group 1 or use additional syringe if needed.

3. For each animal: a. Fill the pipette with the exact dose and pour the liquid to the sublingual area at once (gently) i. Remove the pipette immediately ii. Start the timer per each rat iii. Take photos for record iv. Wake the rat b. Bleeding sampling points (and sampling times) and mean concentration of midazolam in the samples are demonstrated in Fig. 21.

General parameters

The estimated time of the experiments is 4 hours.

Rats will get their usual diet before and during the experiment.

Rats will be under full anesthesia through the residency of the device. Safety parameters are collected through the experiments.

End of the experiment

Blood samples are centrifuged and frozen. Thereafter, the frozen samples are transferred to analytical lab for PK parameters analysis. Statistical analysis is performed in order to calculate Tmax, Cmax, AUC and other parameters which allows concluding the precise residual time required in the experiment. Tables 5A-B below present the dosages administered to each rat in each group. Table 6 provides the sampling points (in minutes) for each rat with the respective midazolam concentration in blood.

Results

1. Raw data from experiments

Tables 5A-5B: Table 5A (Left) describes the initial weight of the device, the weight after soaking (t=0), after 30 min and the administered dose, for each rat of group 1; Table 5B (Right) describes the administered dose by the rats of group 2. 2. Analytics

Table 6: describes the weight of each rat in each group; the sampling times and the mean concentrations of midazolam in blood. *Group 1, Rat #3 showed out of spec results that assumed to be concerned with analytical measurements.

3. Statistics a. Bioavailability measures

Table 7: provides bioavailability measurements results for each rat in each group. b. Mann-Whitney test to compare between the Groups.

Table 8: provides Mann-Whitney test results a. Grouping Variable: group b. Not corrected for ties.

Conclusions

1. Results demonstrate significant difference between the groups.

2. The relative bioavailability is: F=AUC1 (Group #1)/AUC2 (Group #2) = 29565/8988 = 3.28 excluding the out of spec results of Group #1, Rat #3, F=27041/8988=3.01

3. No spillage or drug loss was observed in group #1

4. It is assumed that the loss of material in Group #2 is due to swallowing the formulation

5. The applicator exhibit superiority over existing SL administration, as demonstrated above

6. Onset time seems to be short in both groups, <7 minutes

EXAMPLE 3 - COLORANT COMPATIBILITY TEST

The below presented experiment aim is to observe the absorption of colorants into the buccal tissues via the applicator of the herein invention, positioned in the buccal area by dint of the device.

Assay Specific Information: Ex- Vivo test compatibility colorant. 2 colorants, one with solubility in water and the second one in oil.

Two different Applicator kinds, pre-filled with either oil based green solution or water based blue solution were inserted into the buccal cavity. Specifically: For animal cadaver #829 (immediately following euthanasia), 3 oil-type Applicators coupled each to a reservoir, were inserted under the tongue (1 on each side and 1 in the front) for 60min.

For Animal cadaver #831 (immediately following euthanasia), 6 water-type Applicators, each 3 coupled to a reservoir, were inserted under the tongue (3 on each side of the tongue) for 60min. Results- Following 60 minutes incubation the Applicators were withdrawn and the buccal cavities were pictured:

• Cadaver #829, Colorant Green, oil based: accumulation of liquids was visible in the buccal cavity.

A leak in the connection, between the Applicator to the reservoir of all units was noticed.

• Cadaver #831, Colorant Blue, water based: No liquid was accumulated in the buccal cavity and the tissue under the tongue was stained blue.

Conclusion

Cadaver #829, Green Colorant oil based:

A fade green color was detected under the tongue.

Liquid accumulation in buccal cavity was observed due to reservoir connection point leakage (not from applicator).

Cadaver #829, Trypan Blue water based:

Trypan blue is a vital stain and is absorbed into all cells (alive and dead) but immediately extracted from living cells leaving only dead cells and free fluids stained blue. As the cadaver was fresh the cells were alive and extracted the stain so the blue color accumulated in the free fluids inside the veins - coloring the blood blue (see appendix for vein scheme)