Multi-region oral membranes for treating periodontal and dental diseases or conditions

Abstract

The present invention relates to the field of dental care products for dental, periodontal and / or any other oral conditions, or for cosmetic treatment. It provides an oral film (5), in particular a pharmaceutical oral film, useful for the simultaneous treatment of gums and teeth of a subject, for example for the treatment and / or prevention of periodontal and dental diseases or conditions, or for cosmetic treatment, such as tooth whitening. The invention also provides a method, such as a cosmetic method using the film and a method of making the oral film (5).

Description

[0001] The present invention relates to the field of dental care products for the treatment and / or prevention of periodontal and dental diseases or disorders or for cosmetic treatment. It provides an oral film, in particular a pharmaceutical oral film, which can be used to treat the gums and teeth of a subject simultaneously, for example for the treatment and / or prevention of dental diseases, periodontal diseases and / or any other oral disorders, or for cosmetic treatment such as teeth whitening. The present invention also provides a method, for example a cosmetic method using the film and a method for preparing the oral film.

[0002] Oral films for the preventive, restorative and regenerative treatment of teeth have found wide application in dentistry due to their adaptable mechanical and biological properties, low production costs and ease of processing. They are able to exert a local action, release drugs rapidly, or act as an oral adhesive system retained in the oral cavity for controlled drug release. Oral polymeric films are commonly used in personal care products and over-the-counter drugs.

[0003] A common application of oral films, also known as "patches", is for teeth whitening (for example, US 6,893,629B2, US10,258,546B2, WO 2021 / 224618, EP 3315118A1, US10,849,729B, US 9,782,338B2, US9,149,417B2, EP 3302405 A1, US 6,949,240B2, EP 1696866A1, EP 2934446A1, US 9,668,970 B2).

[0004] Both extrinsic and intrinsic causes can lead to tooth discoloration. For example, those caused by lifestyle habits or foods, such as drinking coffee, tea, wine, carrots, oranges, or tobacco, can leave stains on the tooth surface. Using certain dental care products, such as mouthwashes containing chlorhexidine and cetylpyridinium chloride, or dental materials such as amalgam, especially those containing silver sulfide, can also cause extrinsic tooth discoloration. In addition, certain medications, therapies, or genetic diseases may cause intrinsic discoloration by preventing the normal development of enamel and dentin. Some of these medications that cause tooth discoloration are antibiotics (such as tetracycline and doxycycline) or antihistamines, antipsychotic medications, and antihypertensive medications, or excessive intake of fluoride. Even some infections in pregnant women can affect the enamel development of the baby. Certain medical treatments, such as radiotherapy and chemotherapy for the head and neck, can also cause tooth discoloration. Tooth trauma may also lead to discoloration. Over time, the outer layer of enamel on the tooth wears away with age, exposing the yellow dentin. Dentin also grows with age, thus reducing the size of the dental pulp. As a result, the translucency of the tooth decreases, making it appear darker. Tooth discoloration can be an important aesthetic problem for dental patients. For example, in the UK, approximately 20% of people are dissatisfied with the color of their teeth. In the US, 34% of people seem to be dissatisfied. According to the American Academy of Cosmetic Dentistry, 99% of people believe that their smile is their most important social feature.

[0005] For example, US2004062724A1 discloses a two-layer or multi-layer patch that includes an enamel-adhesive water-soluble polymer layer containing a tooth whitening agent or other active compound and a coating-etchable backing layer that controls the desired residence time.

[0006] US20070178055A1 discloses a dissolvable tooth whitening patch that contains a whitening agent and a water-soluble polymer system. The whitening patch includes additional ingredients, some of which are active agents, to provide additional benefits such as gum health maintenance or treatment. In addition, the whitening patch also contains a variety of reagents, such as antibacterial agents, mineralizing compounds, desensitizing compounds, anticalculus agents, flavoring agents, anti-inflammatory agents, antioxidants, and vitamins.

[0007] US20150125810A1 describes a re-foldable holding device for insertion into a dental site for controlled delivery of at least one intraoral active material. In one embodiment, different surfaces of the device can contact enamel and gingival tissue.

[0008] Whitening products typically contain peroxide-based reagents that are highly reactive and can damage both enamel and irritate the gums.

[0009] In view of this, the inventors have solved the problem of providing an advantageous oral film that is suitable for simultaneously administering an active agent to teeth and gums. It can be used for the topical and controlled delivery of cosmetic and / or pharmaceutical active agents, for example for teeth whitening and / or treatment, prevention or reduction of the incidence of dental caries, tooth decay, mucositis, gingivitis, periodontitis, peri-implantitis or tooth discoloration. Advantageously, it provides greater flexibility in the selection of active agents compared to systems known in the art. The present invention, in particular the subject matter of the claims, solves this problem.

[0010] The present invention provides a film comprising a first surface, the first surface comprising at least one first region containing a first active agent and at least one second region containing a second active agent different from the first active agent,

[0011] wherein the at least one first region is capable of contacting at least one tooth of a subject and the at least one second region is capable of simultaneously contacting the gums of the subject.

[0012] The film of the present invention is a film having a length, a width and a thickness. The thickness has a minimum extension. The thickness of the film can be 1 - 3000 μm, for example 20 - 2000 mm, 40 - 1000 μm, 50 - 800 μm, such as 100 - 500 μm or 100 - 200 μm.

[0013] The first surface generally extends in the length and width of the film. It is the surface on which the first region and the second region are arranged. Generally, the first and second regions are adjacent to each other, i.e., the first region is adjacent to the second region in the first surface.

[0014] The at least one first region is capable of contacting at least one tooth of a subject and the at least one second region is capable of simultaneously contacting the gums of the subject. Thus, the first region can contain at least one active agent suitable for acting on teeth, such as a teeth whitening agent, such as peroxide or a reagent capable of releasing peroxide, an anti-caries agent and / or a desensitizing agent, such as a self-assembling peptide (such as P11-4), and the second region can contain at least one active agent suitable for acting on gums, such as an anti-inflammatory agent, an anti-irritant agent and / or a senescent cell scavenger. For example, the second reagent can include a reagent capable of neutralizing free radicals. In a classic teeth whitening patch containing peroxide or a reagent capable of releasing peroxide, such a reagent would reduce the bleaching activity required for teeth. In the oral film of the present invention, this is not the case because the two reagents are applied in a targeted manner. In addition to the direct incompatibility of the active agents, there are several reagents beneficial for gum treatment that may cause tooth discoloration if applied to teeth, and this discoloration can be avoided or minimized if they are targeted to the gums with the oral film of the present invention. Therefore, the present invention allows for a more targeted treatment of teeth and gums and provides greater flexibility in the selection of active agents.

[0015] The first region of the membrane of the present invention may have dimensions suitable for contacting a single tooth, i.e., the length and width may correspond to the length and width of the tooth. For example, the first region may have an area of 0.5 - 2 cm * 0.5 - 2 cm. Generally, it is suitable for contacting multiple teeth simultaneously. Preferably, it is suitable for contacting at least the labial or vestibular side of the incisors, and optionally the canines. It may also be applicable to contact the labial / vestibular or buccal side of the upper or lower teeth, or the palatal side of the incisors. Optionally, it contacts the incisors and canines, and even contacts the incisors, canines, and all or some of the premolars or molars of the upper or lower teeth, for example. Thus, the first region of the membrane of the present invention may have a length of a single tooth, for example, about 0.5 - 2 cm (e.g., 1 - 1.5 cm) and a width of 3 - 25 cm, such as 4 - 22 cm, 5 - 20 cm, or 7 - 10 cm. It may also be designed to cut a suitable width from a larger membrane, and optionally, a suitable length.

[0016] In one embodiment, the membrane has a first region that not only contacts the labial / vestibular or buccal side of a tooth or teeth but can be mounted around the tooth or adjacent teeth such that the first region contacts both the labial / vestibular or buccal side and the occlusal side (for premolars or molars, the occlusal side). In this case, there may be a second region suitable for contacting the gingiva on the labial, vestibular / buccal side, and optionally, there may be another second region contacting the gingiva on the palatal side (i.e., the opposite side of the first region). For this embodiment, the length of the first region may be about 1 - 4 cm (e.g., 2 - 3 cm), and the width may be, for example, 3 - 25 cm, such as 4 - 22 cm, 5 - 20 cm, or 7 - 10 cm.

[0017] In another embodiment, the membrane may be designed to contact the labial / vestibular side of the upper and lower teeth simultaneously, and optionally, the buccal side. In this case, there may also be two second regions located on the opposite side of the first region.

[0018] Generally, permanent teeth will be treated with the oral membrane of the present invention, but for the treatment of primary teeth, the dimensions may be smaller.

[0019] There may generally be multiple first and second regions, which may also be separated by regions without an active agent, but each first region is adjacent to at least one second region.

[0020] The second region generally extends along the width of the first region. The length of the second region may be, for example, 0.1 to 2 cm, such as 0.2 - 1 cm or 0.3 to 0.5 cm.

[0021] The membrane can be personalized or can be personalized for a specific subject and the shape and size of the subject's teeth. Exact measurements of the membrane or its first and second regions can be selected for a specific subject, typically based on the size of the incisors. If treatment of other teeth is more important, those teeth may determine the size of the membrane used.

[0022] For ease of manufacture and standardization, the shape of the first region is typically rectangular. In this case, due to the curve of the gingival margin, there may be some contact between the first region and the gingiva and / or between the second region and the teeth when applied to the teeth and gingiva. However, compared to membrane treatments that contain only one region, the treatment is generally more targeted overall. The membrane can more specifically conform to the shape of the teeth, particularly the curve of the gingival margin. This allows for a more targeted treatment of the teeth and gingiva.

[0023] The oral membrane can be flat or substantially flat. It can also have at least one recess, at least one protrusion, perforations, or any other type of microstructure. For example, its topography can be configured to physically bind an active agent within a region of the membrane. The height of the region due to such protrusions or microstructures is included within the thickness of the membrane.

[0024] Throughout the invention, the first surface is one surface of the membrane, whether before application or when in contact with a tooth or teeth. There may be an adaptation to the tooth or gingival surface upon contact, but it is not necessary to fold the membrane to contact the subject's teeth and gingiva, as this would change the number of its surfaces.

[0025] In the context of the present invention, the gingiva is defined as the periodontal tissue, i.e., the connective tissue composed of cementum, periodontal ligament, alveolar bone, and gingival tissue. The periodontal tissue surrounds and supports the teeth. Gum is synonymous with gingiva in this application.

[0026] The teeth can be any teeth and, in principle, can be either side of these teeth. Particularly for whitening applications, typically, the labial side of the incisors and optionally, the vestibular side of the canines are in contact with the oral membrane of the present invention, but this can also extend to the buccal side of the molars. For example, in the case of caries treatment or prevention or treatment of periodontitis or aging of the gingiva and teeth, other or additional sides of the teeth can also be contacted. In the context of the present invention, if not specifically mentioned otherwise, the application to teeth includes the application to multiple teeth.

[0027] The subject is typically a human subject, such as an adult or an adolescent. The subject can also be an elderly person, such as a person 50 years old or older. Children can also be treated. The subject can also be an animal, such as a dog, a horse, or a camel.

[0028] The oral film of the present invention is preferably soluble in the oral cavity after the film is applied, particularly when applied to teeth and gums. For example, the film may be substantially dissolved after 1 - 240 minutes, such as 2 - 180 minutes, 3 - 120 minutes, 4 - 90 minutes, 5 - 60 minutes, 8 - 45 minutes, 10 - 30 minutes, or 15 - 20 minutes after application. Preferably, this is the case after 15 minutes or longer to allow sufficient contact time with the active agent. Substantially dissolved means the loss of continuity of the film, with more than 50%, more than 60%, more than 70%, more than 75%, more than 80%, preferably more than 90%, or optionally more than 95% of the film area being dissolved. Of course, the film may also be completely dissolved at this time. Thus, the residence time of the film (i.e., the time until the film is substantially dissolved) is determined by the solubility of the film. The optimal residence time depends on the desired, for example, therapeutic or cosmetic application and / or the active agent in contact with the teeth and gums.

[0029] Optionally, the film comprises a backing layer that forms the second surface of the film opposite the first surface. For example, if the backing layer dissolves more slowly than the material of the first and second regions, such a backing layer can make the residence time longer. If the backing layer impedes the diffusion of one or more active agents, it can also minimize the exposure of the lips or tongue to the active agent in the sheet, which is not only advantageous when one or several active materials are irritants (such as peroxides or reagents capable of providing peroxides), but can also help increase the concentration of the active agent at the teeth and gums, respectively.

[0030] There are different options for the configuration of the oral film of the present invention.

[0031] In one embodiment, the first region is formed by a first sheet and the second region is formed by a second sheet, wherein the surfaces of the first and second sheets together form the first surface. Thus, the film does not have to be a continuous film, but two sheets adjacent to each other can form two regions. This has the advantage that each sheet (forming the first or second region) containing at least one different active agent can be prepared separately. By impregnating an off-the-shelf sheet with at least one active agent, for example, by soaking it in an active agent solution or adding an active agent to the sheet, each sheet can be prepared independently of the other sheet. Alternatively, it can be prepared after mixing the matrix material of the sheet with at least one active agent, such as polymerization, extrusion, or drying.

[0032] When the regions are formed by separate sheets, the residence time of the first region may be different from that of the second region. For example, the first sheet may have a different matrix material and / or different solubility and / or thickness from the second sheet. This may be helpful, for example, if it takes longer to treat the gums than the teeth, or vice versa.

[0033] Optionally, the first sheet or the second sheet may overlap with the respective other region by 10 - 100%. Such an arrangement can simplify the preparation of the film of the present invention. For example, if there is no backing layer, it can firmly fix the two regions in one film. If the overlap is 100%, the structure is particularly strong.

[0034] These sheets may overlap, for example, by 3 - 100%, such as 5 - 50% and 10 - 15%.

[0035] In one embodiment, the first region overlaps with the second region, for example, by at least 3%, such as at least 50% or 100%. Thus, in this case, within the scope of the second region, the first sheet forms a second layer opposite to the first surface of the second region. This has the effect that the active agent contained in the first region can also be provided to the gums by contacting the second region of the teeth. Optionally, in this case, the first region does not contain an irritant. If it contains a reagent beneficial to both the gums and the teeth, such as a self - assembling peptide, such as P11 - 4, this may be advantageous.

[0036] In another embodiment, the second region overlaps with the first region, for example, by at least 3%, such as at least 50% or 100%. Thus, in this case, within the scope of the first region, the second sheet forms a second layer opposite to the first surface of the first region. This has the effect that the active agent contained in the second region can also be provided to the teeth by contacting the first region of the teeth. Optionally, in this case, the second region does not include a reagent that causes problems for the teeth, such as a coloring agent. If it contains a reagent beneficial to both the gums and the teeth, such as a self - assembling peptide, such as P11 - 4, this may be advantageous.

[0037] If the first and second sheets do not overlap, the film of the present invention preferably includes a backing layer to improve stability.

[0038] If the first and second sheets do not overlap, they may be adjacent to each other.

[0039] Thus, they contact each other by at least one of their narrow sides.

[0040] Optionally, the first and second sheets may be separated by a spacer. The spacer may be a region without an active agent. If the film is placed in the oral cavity and / or in contact with water, it is still capable of mediating the diffusion of the active agent. Optionally, the spacer may be configured to inhibit the diffusion of at least one active agent from one region to another region, for example, due to hydrophobicity or the absence of a material. In the latter case, the first surface of the spacer may be formed by a backing layer.

[0041] For example, the spacer can be configured to be placed on the gingival border of a tooth, for example, if it is desired that the active agent in the first region is not applied to the gingiva and / or the active agent in the second region is not applied to the tooth.

[0042] The spacer can be a film extent between at least one first and second sheet. The spacer can be formed by at least one sheet (spacer sheet) disposed between the first and second sheets and adjacent to the narrow sides of the respective sheets. Optionally, the spacer can also surround one or both of the first or second regions.

[0043] Optionally, the spacer sheet can overlap the first and second regions in the first surface, i.e., at the location of the spacer sheet, the first and second regions do not form the first surface.

[0044] In one embodiment, the first region, the second region, and the spacer region (if present) are regions of the same sheet forming the film. Thus, in this case, these regions are all part of a continuous sheet. Thus, these regions are made of a common material. In this case, the film is typically prepared by adding at least two different active agents to their respective regions. To separate the active agents in this embodiment, a spacer region that inhibits diffusion from one region to another may be helpful, for example, by first impregnating the sheet with a hydrophobic material that inhibits the diffusion of hydrophilic active agents.

[0045] For example, the present invention provides a film in which a first sheet and a second sheet forming a first surface are disposed on a backing layer to form a second surface opposite the first surface. The backing layer supports the first and second sheets. If a spacer region is present, the spacer region can be an uncovered region of the backing layer between or around the first and second regions. The spacer region can also overlap the sheets of the first and second regions as described in detail above, or it can be adjacent to the first region on one side and adjacent to the second region on the opposite side.

[0046] Whether or not one or more sheets form the first surface, the material of the backing layer is preferably configured to be soluble in water or saliva. Optionally, it includes cellulose. The backing layer may have a residence time depending on its solubility, which is different from the residence time of the sheet(s) or film(s) constituting the first and second regions depending on solubility.

[0047] Optionally, the backing layer includes at least one third region containing an active agent, where the third region forms at least a part of the second surface. If the first region contacts the tooth and the second region contacts the gingiva, the third region is suitable for contacting the lips and / or tongue of the subject. The third region can include one of the active agents contained in the first or second region, preferably, the same agent as contained in the second region, but optionally or additionally includes a third agent different from the first and second agents.

[0048] Optionally, the backing layer is configured to inhibit the diffusion of at least one active agent into the oral mucosa of the subject. To this end, for example, the water permeability and / or the permeability to the active agent of the backing layer can be lower than that of the one or more sheets forming the first and second regions. Preferably, the residence time of the backing layer is longer than that of the sheets of the first and second regions.

[0049] In the film of the present invention, at least one selected from the group consisting of the backing layer, the spacer, the first region, and the second region can be configured to release ions when placed in the oral cavity. In one embodiment, the backing layer preferably reduces or inhibits the diffusion of the active agent from the second surface of the film into the oral mucosa of the subject and is also capable of releasing ions, such as Figure 3 shown. The release of ions can be triggered, for example, by contact with water or saliva after being placed in the oral cavity. Before being placed in the oral cavity, the film can also be contacted with water to activate it. The release of ions can be used to locally change the pH value. For example, by releasing H + ions, the pH value can be made more acidic. For example, this may be helpful if it is desired to provide the assembled form of the self-assembling peptide to a tooth or teeth, wherein the self-assembling peptide assembles at an acidic pH value, such as for P11-4.

[0050] Optionally, the pH value can be made more alkaline by releasing OH - ions. For example, this may be helpful if it is desired to provide the assembled form of the self-assembling peptide to a tooth, wherein the self-assembling peptide assembles at an alkaline pH value, such as for P11-8. The responses of other components to the teeth and gums may also be caused by the ions released from the backing layer.

[0051] Using the same principle, it can be an additional intermediate layer capable of releasing ions instead of the backing layer, wherein the backing layer preferably reduces or inhibits the diffusion of the active agent from the second surface of the film into the oral mucosa of the subject. The intermediate layer can be located between the backing layer and the one or more sheets forming the first surface of the film.

[0052] Optionally, the film of the present invention, such as the backing layer, includes a battery element, which can be a biological battery or at least one other Galvanic cell. The biological battery converts the chemical energy from an energy carrier, usually a sugar such as glucose or maltodextrin, preferably glucose. It requires a suitable energy carrier, an enzyme capable of converting it, and water. It can be activated, for example, by adding glucose, or by adding water (or saliva) to the dry biological battery, wherein, for example, water (such as from saliva) can enable the energy carrier to diffuse into the biological battery. The enzyme can be contained in a suitable bacterium.

[0053] Other galvanic cells include, for example, nano - batteries. For example, lithium batteries can be used.

[0054] A battery, such as a bio - battery, is preferably a primary battery comprising at least two electrodes separated by a membrane. The bio - battery consists of an anode, a cathode, a separator, and an electrolyte, with each component laminated on top of another. At the anode, sugar is oxidized, generating electrons and protons. The electrons travel from the anode surface through an external circuit to the cathode. On the other hand, the protons are transferred through the electrolyte across the separator to the cathode side of the battery.

[0055] The battery core can be planar. Multiple battery cores can be arranged adjacent to each other in a planar manner, for example, to optimize the transfer of ions and active agents to multiple teeth and / or the gums adjacent to multiple teeth. The battery advantageously optimizes the dose delivery of at least one active agent to at least one tooth, gum, and / or lip or tongue of a subject.

[0056] The battery can be embedded in an intermediate layer or preferably in a backing layer. The bio - battery can include at least one, optionally two, planar mesh electrodes within the embedding layer. If the battery is a bio - battery, a preferred option is an embedded enzyme bio - battery. The layers holding the battery are generally non - degradable, but the layer containing the first and second regions may be degradable. Thus, it allows for better delivery of the active agent through the first surface to the gums and / or teeth. As Figure 2 shown, the battery can act on the first and second regions or on the first or second region. It can be embedded in the backing layer and act on the first and second regions or on the first or second region. Optionally, it can be placed on the back of one or more sheets holding the first and / or second regions as an intermediate layer.

[0057] Optionally, instead of a battery, at any position where a battery is described, the membrane of the present invention can include electrodes that can be connected to an external battery and can be used in combination with another external electrode to provide controlled ion transport to the tooth surface and / or gums.

[0058] In the context of the present invention, different sheets or layers can be connected to each other by physical (e.g., network interpolation or hydrophobic interaction, ionic interaction) and / or chemical interactions (e.g., covalent bonds). They can be, for example, pressed, welded, thermally welded, laser welded, ultrasonically welded, 3D printed, glued, coated, cast, or chemically connected, such as cross - linked to each other.

[0059] The oral membrane or at least one of its sheets preferably includes a polymer material and / or a hydrogel as a matrix material. The matrix material is preferably water - soluble, so it dissolves when it comes into contact with saliva. They are also non - toxic and preferably do not interact with the active agent to the extent that it prevents its effect on the teeth or gums.

[0060] The matrix material may include vinyl polymers such as polyvinyl acetate (PVA), polyvinylpyrrolidone (PVP), polyacrylates, PEEK (polyetheretherketone) polymers, carbomers, natural polymers or their copolymers, or pectin-coated liposomes. For example, the polymeric material may include natural polymers such as cellulose-based polymers such as hydroxypropylmethylcellulose (HPMC), hydroxypropylcellulose (HPC), ethylcellulose (EC), carboxymethylcellulose (CMC) or chitosan, collagen, gelatin, silk, alginate, xanthan gum, albizia gum or Khaya gum. HPMC is a particularly preferred natural polymer.

[0061] The polymeric material may additionally contain one or more plasticizers such as glycerol or PEG such as PEG-400.

[0062] The polymeric material may for example include PVP, for example without plasticizer or containing 1 - 30% plasticizer. Low or medium amounts of plasticizer, such as 0 - 15%, 1 - 12%, 3 - 10%, 4 - 8% or 5 - 6% plasticizer (preferably glycerol) have been found to be advantageous. In a preferred embodiment, the matrix polymer comprises 95% PVP and 5% glycerol or PEG-400, preferably glycerol. The percentages herein relate to the mass of the polymer and the plasticizer (but not to the active ingredient).

[0063] The polymeric material may for example include PVA, for example without plasticizer or containing 1 - 30% plasticizer. Low or medium amounts of plasticizer, such as 0 - 15%, 1 - 12%, 3 - 10%, 4 - 8% or 5 - 6% plasticizer (preferably glycerol) have been found to be advantageous. In a preferred embodiment, the matrix polymer comprises 95% PVA and 5% glycerol or PEG-400, preferably glycerol.

[0064] The polymeric material may for example include HPMC, for example without plasticizer or containing 1 - 30% plasticizer. Low or medium amounts of plasticizer, such as 0 - 15%, 0 - 12%, 0 - 10%, 0 - 8% or 0 - 6%, 0 - 5%, 0 - 3% or 0 - 1% plasticizer (preferably without plasticizer) have been found to be advantageous. In a preferred embodiment, the matrix polymer comprises 100% HPMC and no plasticizer.

[0065] The polymer can be stabilized by chemical and / or physical crosslinking. It can also be an aerogel of self-assembling peptides such as P11-4 or P11-8, or a combination of at least one of the above polymers with self-assembling peptides. Additional ingredients can be added to enhance matrix properties (such as dopa (3,4-dihydroxyphenylalanine)).

[0066] Natural polymers combined with vinyl polymers have proven to have advantageous properties as matrix materials (e.g., mixtures of PVA and HPMC or PVP and HPMC, preferably PVP and HPMC).

[0067] Hydrocolloids can encapsulate, protect, and release active agents in a controlled manner, e.g., for wounds in the gingival or oral region. Biocellulose, i.e., cellulose-based polymers, may contribute to periodontal regeneration and has been successfully used for dressing oral mucosal surgical wounds.

[0068] At least two regions of the membrane can comprise the same or different polymer matrices. Different polymer matrices are particularly needed if different dissolution and release characteristics are required, e.g., as shown in the examples below.

[0069] The material of the membrane is preferably at least partially flexible such that the membrane or at least the first and second regions can be respectively disposed in the periodontal or dental treatment area, particularly the gingiva, attached gingiva, mucosa, mucogingival junction, gingival sulcus, and / or interdental papilla as the free margin of the periodontal treatment area, and the facial, lingual, incisor, and / or occlusal tooth surfaces as the dental treatment area. The membrane can, for example, comprise at least one flexible region, where the flexible region can be a spacer region. The membrane can also comprise less flexible or non-flexible rigid regions, e.g., one or more of the first, second, or third regions. Flexibility of the membrane is preferred as it can conform well to the teeth and gums, thus transferring the active agent well. Optionally, the membrane of the present invention can have a rigid spatial shape such that at least one first region and at least one second region respectively conform to the periodontal or dental treatment area of the subject. Typically, in such cases, it is personalized.

[0070] Regardless of the flexibility or rigidity of the membrane or its regions, the membrane is not designed to rest and be applied on the sulcular epithelium, gingival sulcus, junctional epithelium, periodontal ligament, or cementum as the treatment area.

[0071] In addition to the matrix material and the active agent, the membrane or sheet comprising at least the first and second regions can also comprise other components, such as plasticizers, sweeteners, flavoring agents, surfactants, saliva stimulants, permeation enhancers, buffers, and / or mucosal or tooth adhesion enhancers.

[0072] In one embodiment, the sheet comprising the first and / or second (and, optionally, the third region) can comprise

[0073] · 1 - 30 wt% of a natural polymer, e.g., 3 - 10 wt% or 5 - 12 wt%, e.g., about 9%;

[0074] · 0 - 30 wt% of a copolymer (e.g., a vinyl polymer), e.g., 1 - 10 wt% or 5 - 10 wt%;

[0075] · 0 - 10 wt% of a plasticizer, such as 1 - 5 wt%;

[0076] · 0.1 - 50 wt% of an active agent, such as 1 - 40, 2 - 30, 3 - 25, 4 - 20, 4 - 15 or 5 - 10 wt%;

[0077] · 0 - 5 wt% of a sweetener, such as 1 - 2.5 wt%;

[0078] · 0 - 10 wt% of a flavoring agent, such as 1 - 5 wt%;

[0079] · 0 - 5 wt% of a surfactant, such as 1 - 3.5 wt%;

[0080] · 0 - 5 wt% of a saliva stimulant, such as 1 - 2.5 wt%, and optionally;

[0081] · Water to make up 100 wt%.

[0082] If the active ingredient is a self - assembling peptide, it is preferred to use a buffer instead of water to stabilize the peptide in monomer form. The inventors were able to demonstrate that, despite the drying process, the self - assembling peptide in the film remained in the monomer state in different polymer matrices.

[0083] The membrane or at least its first surface is at least partially sticky, i.e., at least one area of said surface, preferably the entire first surface, is sticky such that during the residence time of the membrane, it can hold the membrane on at least one tooth and the adjacent gingiva. The membrane can be held on the tooth and / or gingival surface by the adhesive action of the matrix material of the oral membrane, preferably without the need for an additional adhesive layer that does not contain an active agent. If necessary, the membrane can also be designed to adhere to crowns, bridges, and veneers. The adhesion can be through physical interaction or through mixed-bond adhesion. The sheets of the first and second regions, and optionally the spacer regions, can optionally include adhesives of synthetic or natural origin, such as polyacrylic acid and / or polyvinylpyrrolidone complexes. Different topographies of the surface may also contribute to adhesion, such as those disclosed by Bovone et al., 2021. ACS Biomater Sci Eng. 2021;7(9):4048 - 4076. Doi:10.1021 / acsbiomaterials.0c01677; Yang et al., 2020. Adv. Funct. Mater. 2020, 30, 1901693. https: / / doi.org / 10.1002 / adfm.201901693; or Pelaez-Vargas et al., 2010, Microsc Microanal. 2010;16(6):670 - 676. Doi:10.1017 / S1431927610094158.

[0084] In the context of the present invention, the active agent is not limited to the active agents considered as such in the corresponding market or marketing authorization, which will vary according to the type of product and the applicable legislation, but rather any compound having a cosmetic, therapeutic, prophylactic, or preventive effect on the teeth and / or gingiva.

[0085] In the context of the present invention, the first active agent can be selected from peroxides, reagents capable of generating peroxides, self-assembling peptides, CPP-ACP (casein phosphopeptide-amorphous calcium phosphate), mineral particles such as hydroxyapatite, amorphous calcium phosphate, and fluorides. Combinations thereof can also be used and are indeed preferred, for example, using at least two or at least three active agents. Unless otherwise explicitly stated, a reference to an active agent refers to a reference to at least one active agent. The first active agent is contained in the first region of the membrane, which is intended to be applied to the teeth of a subject. Thus, the first active agent is suitable for treating teeth. For example, it can be suitable for teeth whitening, caries treatment, caries prevention, and / or desensitization of sensitive teeth.

[0086] There are various tooth whitening techniques known, but most are based on bleaching with oxidizing agents such as peroxides. In particular, most tooth whitening products on the market use hydrogen peroxide (HP). However, in recent years, there has been an increased awareness of bleaching-related side effects such as demineralization, erosion, and tooth sensitivity caused by peroxides (e.g., Dahl et al., 2003, Crit Rev Oral Biol Med 14(4):292-304). Peroxides are also irritating to the gums and can cause inflammation. Other peroxides, such as carbamide peroxide (CP) or reagents capable of generating peroxides, are considered to have milder effects. For example, a combination of HP and CP can be used, or phthalimido peroxyhexanoic acid (PAP) can be used alone or in combination with HP. If a peroxide or a reagent capable of providing a peroxide is used in the oral film of the present invention, PAP is preferred.

[0087] Self-assembling peptides (SAPs) are known for their excellent properties in the treatment and prevention of carious lesions (e.g., WO2004 / 007532A1). WO 2004 / 007532A1 discloses peptides capable of forming three-dimensional scaffolds that promote de novo calcium phosphate nucleation. These artificial peptides assemble in one dimension to form β-sheets and higher-order assemblies such as ribbon assemblies. A three-dimensional supramolecular structure of SAP can be formed, which has an affinity for calcium phosphate.

[0088] In the context of the present invention, self-assembling peptides taught in WO 2004 / 007532A1, US10 / 521628, US12 / 729046, US13 / 551878, US14 / 062768, or WO 2014 / 027012A1 are preferred, and these are hereby incorporated by reference in their entirety. In particular, self-assembling peptides with a net charge of +2 or -2 at pH 7.5 can be used in monomeric or assembled form.

[0089] The self-assembling peptides used in the present invention preferably comprise a sequence of the formula X1-X2-X1-X2-X1, where X1 is an amino acid with an acidic side chain and X2 is an amino acid with a hydrophobic side chain. For example, the self-assembling peptides used in the present invention comprise the consensus sequence SEQ ID NO:1, X1-X2-X1-X2-X1, where X1 is independently selected from glutamic acid, aspartic acid, glutamine, and ornithine, and X2 is independently selected from alanine, valine, isoleucine, leucine, methionine, phenylalanine, tyrosine, tryptophan, and glutamine, or an amino acid sequence having at least 80% identity therewith. Independent selection means that, for example, X1 at positions 1, 3, or 5 of the above sequence can be different from each other. Of course, they can also be the same.

[0090] Preferably, the self-assembling peptide used in the present invention comprises SEQ ID NO:2, X1-X2-X1-X2-X1, wherein X1 is independently selected from glutamic acid and ornithine, and X2 is independently selected from tryptophan and phenylalanine.

[0091] In another embodiment, the self-assembling peptide used in the present invention may comprise SEQ ID NO:3, X3-F-X1-W-X1-F-X1, wherein X1 is independently selected from glutamic acid and ornithine, X3 is selected from arginine, glutamic acid and ornithine, and wherein X3 is preferably arginine.

[0092] In addition, the self-assembling peptide used in the present invention may contain or preferably consist of SEQ ID NO:4: X4-X4-X3-F-X1-W-X1-F-X1-X4-X4, wherein X1 is independently selected from glutamic acid and ornithine, wherein X3 is selected from arginine, glutamic acid and ornithine, and wherein X4 is independently selected from glutamine, glutamic acid, serine, threonine and ornithine. X3 is preferably arginine. Independently, X4 is preferably glutamine.

[0093] The self-assembling peptide of the present invention may contain or preferably consist of SEQ ID NO:5: Q-Q-R-F-X1-W-X1-F-X1-Q-Q, wherein X1 is independently selected from glutamic acid and ornithine.

[0094] Preferably, the self-assembling peptide used in the present invention comprises or consists of a sequence selected from the consensus sequences listed in Table 1 below.

[0095] Most preferably, the peptide comprises or consists of the specific peptides listed in Table 2. Of course, as disclosed herein, a self-assembling peptide in combination with another self-assembling peptide may be formulated in a kit or a composition.

[0096] Peptides of SEQ ID NO:6, 9, 11, 12, 16 or 17 are particularly advantageous, for example, because they can be used at relatively low concentrations, they are highly compatible with cells and have a beneficial charge distribution.

[0097] Preferably, the self-assembling peptide comprises or consists of the sequence of SEQ ID NO:6. The peptide consisting of the SEQ ID NO:6 sequence is also designated as P11-4 and is preferred in the present invention. In another preferred embodiment, the self-assembling peptide comprises or consists of the sequence of SEQ ID NO:9 (P11-8).

[0098] The kit or composition of the present invention may further comprise at least one self-assembling peptide having at least 45% sequence identity with the peptide consisting of SEQ ID NO:6. Preferably, the peptide has at least 54%, at least 63%, at least 72%, at least 81% or at least 90% sequence identity with the peptide consisting of SEQ ID NO:6 or is the peptide. The peptides of the present invention are, for example, 11 amino acids in length.

[0099] The self-assembling peptide may be a modified peptide comprising an Ac-N-terminus and / or an NH2-C-terminus, preferably both, or an unmodified peptide. Since the non-closed form tends to start the deamination reaction, it is preferred to block the ends of all self-assembling peptides of SEQ ID NO:1 to improve stability. In particular, the peptides of SEQ ID NO:6, 9, 11, 12, 16 and 17 may comprise an Ac-N-terminus and an NH2-C-terminus. SEQ ID NO:18-29 correspond to the modified peptides of the present invention.

[0100] Table 1: Consensus sequence of preferred self-assembling peptides

[0101]

[0102]

[0103] Table 2: Preferred self-assembling peptides. Position X1 is underlined

[0104]

[0105] The self-assembling peptides preferably do not have cleavage sites for endopeptidases in the subject. They also do not need to contain special recognition motifs for cells.

[0106] The mineral particles are preferably calcium phosphate particles, such as hydroxyapatite particles. As described in WO 2015 / 044268 A1, a composition comprising 0.4-40 wt% of mineral particles, preferably hydroxyapatite particles with a size of 0.01-50 μm and 0.001-5 wt% of a self-assembling peptide (such as the peptides described herein, preferably P11-4) also has excellent tooth whitening effects.

[0107] Other suitable mineral particles are amorphous calcium phosphate (ACP). ACP, for example in the context of CPP-ACP, has also been shown to contribute to tooth remineralization. WO 2022 / 084288A1 shows that the combination of SAP and ACP can accelerate the remineralization of SAP. Reagents capable of generating ACP can also be used as active agents.

[0108] It has now been recognized that peroxides or reagents capable of generating peroxides, such as PAP and SAP, can be advantageously combined, optionally in combination with the mineral particles disclosed in WO 2015 / 044268A1. Accordingly, the present application provides an oral care product comprising a peroxide or a reagent capable of generating a peroxide, such as PAP and SAP, optionally in combination with these mineral particles. Such an oral care product, such as the oral film of the present invention, which comprises these reagents in a first region, can be used not only to improve tooth whitening effects, but also to avoid demineralization, corrosion and tooth sensitivity usually caused by peroxides. It may also contribute to the remineralization of teeth.

[0109] Fluoride can also improve the remineralization of teeth, because the presence of fluoride in tooth enamel makes it more difficult for teeth to remineralize. The fluoride can be, for example, sodium fluoride or sodium monofluorophosphate with a fluoride content of up to 1500 ppm. The fluoride can be advantageously combined with a combination of SAP and an active agent comprising SAP, for example, to improve caries prevention, i.e., to prevent or reduce the incidence of dental caries.

[0110] A second region capable of contacting the subject's gums comprises at least one active agent different from the first active agent. However, it may additionally comprise at least one active agent that is also present in the first region, i.e., the first active agent. The second active agent may be selected from anti-inflammatory agents, senescent cell scavengers, plant preparations, antibacterial agents and self-assembling peptides.

[0111] In tooth whitening, for example involving peroxides, a common problem is the irritation and inflammation of the gums near the treated teeth caused by peroxides. This may also lead to an increase in tooth sensitivity. It further causes gum aging. Therefore, it is advantageous to counteract any such risks while treating the teeth.

[0112] The anti-inflammatory agent can be, for example, a plant preparation or a metal agent, such as gold particles, or a non-steroidal anti-inflammatory drug (NSAIDs). Typical senescent cell scavengers are also plant preparations, usually colored substances, which may cause tooth staining if applied to the teeth. However, with the oral film of the present invention, such potential tooth staining agents can also be used to target the treatment of the gums.

[0113] The antibacterial agent can also be a plant preparation. They can also be antibiotics.

[0114] Table 3 provides exemplary suitable plant preparations and their main therapeutic effects. It should be noted that both isolated active agents and plant extracts can be used as active agents, such as water extracts, ethanol extracts, methanol extracts, oils, such as sunflower oil extracts, propylene glycol extracts, PEG extracts, hexane extracts, CO2 extracts or glycerol extracts or dry extracts. In the context of the dental care products of the present invention, it may be desirable to provide a product that does not contain ethanol, methanol, hexane, PEG or propylene glycol. Therefore, glycerol extracts or dry extracts, such as glycerol extracts, can be advantageously used. Glycerol extracts can also contain water. Of course, combinations of plant preparations can also be used.

[0115] Table 3:

[0116]

[0117]

[0118] References for aloe vera juice: Indian Journal of Oral Sciences (Vol. 3(2) 2012.; Geetha Bhat et al., J Indian Soc Periodontol. 2011 15(3): 205-209.

[0119] References for neem extract: IOSR Journal of Pharmacy Vol 3(4), 15 / 21 2013

[0120] References for turmeric: Gen Dent 2012 60(5): e283 / 7 K Indian Soc Periodontol 2011, 15(·1): 35 = 38

[0121] References for propolis: Journal of Pharmaceutical Sciences Vol 96(8), 2074 / 2089, 2007

[0122] Alternative active plant preparations are extracts from plants of the genus Rhododendron, preferably plants of the genus Rhododendron L classified by Chamberlain 1996. Advantageously, the extracts are from Rhododendron ferrugineum (sometimes called alpenrose or alpine rhododendron, snow rose or rusty-leaved alpenrose) or Rhododedron hirsutum, as described in US11154470 B1.

[0123] It has been shown that this extract has advantageous characteristics, in particular its ability to

[0124] · inhibit UVA-induced photoaging (protein carbonylation)

[0125] · exhibit anti-aging effects, i.e., it eliminates senescent cells without having a negative impact on healthy cells. This rejuvenates the entire tissue as senescent cells cause inflammation and aging.

[0126] · significantly reduce redness and swelling and increase elasticity.

[0127] The extract also has a potent antibacterial effect, for example against bacteria in periodontal disease.

[0128] Dental care products can contain, for example, 0.1 - 5 wt%, for example 0.5 - 3 wt% of the said extract. It has been found that using 1 - 2 wt% is advantageous. The extract can be, for example, a leaf extract extracted at a drug-to-extract ratio of 1:10 to 1:100, preferably about 1:50, for example extracted with glycerol.

[0129] The inventors have found that it is particularly advantageous to use the dental care product of the present invention in combination with an extract of plants of the genus Leontopodium, the dental care product containing an extract of plants of the genus Rhododendron. Possibly due to its high content of antioxidants, the extract has anti-inflammatory properties and advantageously synergistically treats the gums of a subject with the rhododendron extract, as further explained below, for example treating gingivitis, periodontitis and peri-implantitis, and regenerating the gums.

[0130] Thus, the dental care product also contains 0.1 - 5 wt% of an extract of plants of the genus Leontopodium, preferably Leontopodium alpinum, such as Leontopodium nivale, which is also named Edelweiβ, as described in US 11154470B1.

[0131] Dental care products can, for example, contain 0.5 - 3 wt% of the said extract. It has been found that using 1 - 2 wt% is advantageous.

[0132] It has antioxidant and anti-inflammatory properties. The adsorption and elimination of free radicals are improved, which helps to combat the aging of the gums. The high tannin content also has a protective effect. Tannins can also increase elasticity and contribute to the regeneration of the gums. Flavonoids protect the vascular system. Phenylpropanoids have an astringent effect. It stimulates blood circulation and improves cell activity, and thus, in particular, in combination with the active agents in the rhododendron extract, stimulates the regeneration of tissues.

[0133] Optionally, the dental care product of the present invention further comprises an extract of the genus Eleutherococcus, optionally Eleutherococcus senticosus, commonly known as Siberian ginseng, as described in US11154470 B1. It has anti-inflammatory, anti-irritant and anti-stress activities and can improve wound healing. The dental care product of the present invention may contain, for example, 0.1 - 5 wt% of the extract of the genus Eleutherococcus, such as 0.5 - 2 wt% of the above-mentioned extract. Usually, if the extract is present, it contains 1 - 2 wt%. The extract may be, for example, a root extract extracted at a drug-to-extract ratio of 1:10 to 1:100, preferably about 1:50, for example, extracted with glycerol.

[0134] Preferably, the dental care product does not contain ethanol. Thus, in a preferred embodiment, all the herbal extracts contained are non-alcoholic extracts, such as glycerol extracts, oil extracts or dry extracts. Such glycerol extracts may also contain other ingredients or solvents, preferably water, but they do not contain ethanol. The dental care product of the present invention can be halal and / or vegan.

[0135] Preferably, in the oral film of the present invention, at least the first region contains the self-assembling peptides disclosed herein, such as SAPs containing SEQ ID NO:1 or having at least 70% sequence identity therewith, or P11-4 of P11-8. Preferably, the SAP is P11-4. The concentration range of the SAP can be, for example, 5 - 50 mg / mL. The second region may also include the SAP. In this case, the first region and the second region further include different active agents. For example, the first active agent may be a peroxide or a reagent capable of generating a peroxide, such as HP / CP or PAP, preferably PAP. Its concentration range can be, for example, 2 - 20 wt%. Optionally, the first region further includes fluoride and / or minerals, such as calcium phosphate particles having the size disclosed herein, making it suitable for tooth whitening. The second active agent is preferably an anti-inflammatory plant preparation, as described above. Alternatively or additionally, the second active agent may be a prebiotic to balance the oral microbiome, such as inulin and / or Vaccinium macrocarpon fruit extract, for example, at a concentration range up to 5 wt%.

[0136] The oral film of the present invention can also be a pharmaceutical film, that is, it can be used for treating and / or preventing (e.g., reducing the incidence of) at least one disease or disorder. In particular, the oral film of the present invention can be used for treating or preventing at least one periodontal or dental disease or disorder selected from dental caries, mucositis, gingivitis, periodontitis, peri-implantitis and tooth discoloration in a subject. Preferably, it is used for treatment.

[0137] The present invention also provides a method of treating a subject using the membrane, preferably a cosmetic method, comprising contacting at least one tooth of the subject with a first region of the membrane and contacting at least one region of the subject's gingiva with a second region of the membrane. After administration, the presence and flow of saliva may activate the membrane, i.e., the active agents from different regions start to contact the corresponding target regions (tooth or gingiva), for example, by diffusion. By providing ions or using a bio-battery as disclosed herein, the active agents can also be actively propelled onto or into the target tissue. If the membrane or a sheet thereof of the present invention is water-soluble, the dissolution of the membrane also contributes to the delivery of the active agent. The action of the active agent can last, for example, from 15 minutes to one hour or several hours, depending, for example, on the solubility of the membrane or a sheet thereof, until the membrane is finally dissolved. A backing layer having a lower or no permeability to the active agent and / or a lower solubility compared to the sheet forming the first and second regions may contribute to targeted delivery.

[0138] Preferably, only the tooth contacts the first region. For example, for the treatment of upper teeth, the first region adheres to the Figure 4 tooth below the line shown, and the second region (or spacer, if any) adheres above the line, where the second region mainly contacts the gingiva.

[0139] The method may also include removing the undissolved membrane or its residue. Of course, if the membrane or a part thereof is insoluble, it needs to be removed. The removal can be by manually removing the membrane, or, for example, in the case where the membrane is partially dissolved, washing it off with water or a suitable solvent, which may optionally be provided with the membrane in the kit. For example, if the membrane is more soluble in ethanol than in water, an ethanol mouthwash may be provided with the membrane in the kit to wash off the membrane or any of its residues after the desired administration time. The mouthwash may of course contain active agents, such as anti-inflammatory agents and / or reagents capable of neutralizing any remaining peroxide.

[0140] The present invention also provides a method of preparing the membrane of the present invention, comprising:

[0141] providing a membrane comprising a first surface, the first surface comprising at least one first region and at least one second region and adsorbing at least a first active agent in the first region and at least a second active agent in the second region. When adding the active agents, the backing layer is generally already attached to the sheet comprising the first and second regions.

[0142] However, preferably, the method of preparing the membrane of the present invention comprises preparing a first sheet by mixing a matrix material of the first sheet with at least a first active agent and preparing a second sheet by mixing a matrix material of the second sheet with at least a second active agent.

[0143] The method may include co - casting. Optionally, in the first step of the polymerization process, physical separation prevents mixing of the formed polymer matrix, and in the second step, the physical separation is removed to allow the bonding of the two regions, which minimizes the mixing of the components and / or different matrix materials (if applicable).

[0144] Optionally, the method includes attaching part or all of one of the first and second sheets to the other to form a first surface, where the sheets overlap by 3 - 100%, optionally 10 - 50%. Optionally, the method includes attaching a spacer between the first and second regions, where the spacer is adjacent to these regions. The method may also include further attaching spacers on the first and second regions, where the spacers overlap these regions by 10 - 50% respectively.

[0145] The membrane is assembled from the first and second sheets and the spacer (if present) and optionally a backing layer. If there is an intermediate layer, for example in the case of a bio - battery, the membrane is assembled from an intermediate layer between the first and / or second sheets and the backing layer.

[0146] If present, a third region may be attached to the second surface of the backing layer opposite the first surface.

[0147] Optionally, the method of preparing the membrane of the present invention involves three - dimensional printing the membrane in a personalized manner, taking into account the size and shape of the tooth, and optionally the condition or disorder of the subject and its severity. This allows for the most precise targeting of the active agent, as the shape of the gingival border can be reflected by the boundary between the first and second regions.

[0148] The method of preparing the membrane of the present invention may also optionally include packaging the membrane. Description of the Drawings

[0149] Figure 1 Different embodiments of the membrane of the present invention are shown. The different components are slightly shifted laterally, which is only for better observation in the figures. In A, the first sheet 10 including the first region 1 overlaps the second sheet 20 including the second region 2 by 100%. In B, the first sheet 10 including the first region 1 and the second sheet 20 including the second region 2 are adjacent to each other and are located on the backing layer 30. In C, the first sheet 10 including the first region 1 and the second sheet 20 including the second region 2 are adjacent to each other, but at the boundary between these sheets, the spacer 3 overlaps a part of the first region 1 and / or the second region 2. D shows the same configuration as A, but with a backing layer 30. E shows the same configuration as C, but with a backing layer 30. In A, the first surface 4 is shown with an arrow. F shows an embodiment where the first region 1 and the second region 2 are located on a continuous sheet 10 / 20 and there is a backing layer 30.

[0150] Figure 2 shows a dual - zone multilayer film 5 in which a biobattery 31 is embedded Figure 2 A a backing layer 30 behind the first and second zones 1, 2 on sheets 10 and 20, Figure 2 B behind the second sheet 20, with a separate biobattery in an intermediate layer 40, placed Figure 2 C on the back side of sheet 20. The back or "rear" side means the side opposite the first surface 4. Sheet 20 is shown as transparent, which is not necessary.

[0151] Figure 3 shows a dual - zone multilayer film 5 with a backing layer 30 having ion impermeability or low ion permeability. The active agent shown in zone 2 / sheet 20 is shown to be provided to the gums by forced ion diffusion.

[0152] Figure 4 Preferably, only the tooth contacts the first zone 1 of the film 5 of the present invention. For example, for treating the upper teeth, the first zone 1 adheres to the tooth below the shown line, and the second zone 2 (or spacer 3, if present) adheres above the line, where the second zone 2 mainly contacts the gums.

[0153] Figure 5 Bioadhesion data of market samples, placebo, and loaded films

[0154] Figure 6 Dissolution times and corresponding thicknesses of the selected films

[0155] Figure 7 Release curves and corresponding film thicknesses of three films

[0156] Figure 8 An aspect of films made of HPMC, PVP, and an 8:2 mixture of both. B Side - by - side oral films and a single matrix.

[0157] 1 First zone

[0158] 2 Second zone

[0159] 3 Spacer

[0160] 4 First surface

[0161] 5 Film

[0162] 6 Second surface

[0163] 10 First sheet

[0164] 20 Second sheet

[0165] 30 Backing layer

[0166] 31 Battery, such as a bio - battery

[0167] 40 Intermediate layer Example

[0168] A proof - of - concept study was conducted to evaluate the feasibility of incorporating peptide P11 - 4 into a membrane formulation that uses a matrix - forming polymer to assess its release. The goal was to achieve the release of P11 - 4 within 30 minutes and complete dissolution of the membrane approximately 45 minutes later - something that existing products cannot achieve. Additionally, the membrane should prevent premature aggregation of self - assembling peptides such as P11 - 4. The possibility of preparing a dual - matrix membrane, which consists of two independent regions with different properties and can be used to deliver active molecules with different performances, was also evaluated. Dual - matrix membranes could be targeted, for example, in oral applications because they can treat both teeth and gums simultaneously.

[0169] Laboratory - scale film - casting methods were tested on selected compositions. Dual - matrix membranes in side - by - side configurations were also prepared. By developing analytical methods, candidate formulations were characterized, ranked, and compared with commercial products. These analyses were able to evaluate in vitro the key physicochemical properties of individual matrices, such as bioadhesion, peptide aggregation within the membrane, dissolution time, and peptide release kinetics.

[0170] This work shows that, for the first time, using common US Food and Drug Administration - approved pharmaceutical excipients, oral membranes containing peptide P11 - 4 can be formulated without signs of aggregation. Leading membranes have bioadhesive properties comparable to the marketed alternatives provided, while their dissolution and release characteristics are within the target range. The different properties observed between the matrices mean that the preparation of dual - matrix dosage forms allows for targeted multimodal release profiles.

[0171] The results of this study indicate that oral membranes with satisfactory properties can be formulated.

[0172] 1. Materials and methods

[0173] 1.1. Composition considerations

[0174] The composition of a benchmark commercial whitening patch (Table 4) shows that dental whitening membranes are typically a dual - polymer mixture containing one or more plasticizers and other excipients such as buffers or flavoring agents. Using mixtures can improve the performance of the membrane; however, the efficacy depends largely on the polymer characteristics, mixing ratios, and plasticizers. A common feature in all commercial formulations is polyvinylpyrrolidone (PVP) of unknown molecular weight. It is a hydrophilic, biocompatible synthetic polymer that has been reported to have strong mucoadhesive properties (Jain et al., 2008. Pharmazie. 2008, No. 2, 129 - 135).

[0175] Table 4: Benchmark composition

[0176]

[0177] 1.2. Materials

[0178] The following film-forming polymers were used in this study: hydroxypropyl methylcellulose - HPMC (Dow Chemical Company, Midland, Michigan, USA), polyvinyl alcohol 72000 biochemical - PVA (Applichem, Darmstadt, Germany), polyvinylpyrrolidone K90 - PVP (Fluka, Sigma - Aldrich Chemie, Steinheim, Germany). As plasticizers, two compounds were used: anhydrous glycerol (Fluka, Sigma - Aldrich Chemie, Steinheim, Germany) and polyethylene glycol 400 - PEG 400 (Applichem, Darmstadt, Germany).

[0179] For the buffer solution and release medium, Dulbecco's phosphate - buffered saline - DPBS (Sigma - Aldrich Chemie, Steinheim, Germany) and sodium phosphate dihydrate (ACROS, Geel, Belgium) were used. The xanthan gum (NOW Foods, Bloomingdale, Illinois, USA) water had MiliQ quality.

[0180] 1.3. Preparation of membranes

[0181] Blank membranes were cast by solvent evaporation. Briefly, at 80 °C with magnetic stirring, a 9 w% solution of deionized water containing the polymer and a certain amount of plasticizer (as described in Table 2 below) was dissolved for 24 hours to fully hydrate the polymer. The solution was poured into a borosilicate crystallizer, and the membrane was dried in an active air stream for at least 48 hours. After collection, the membranes were stored in sealed bags at 2 - 5 °C until cut into the desired size and shape before use.

[0182] For the membranes loaded with P11 - 4 peptide, the preparation method was modified from that of the blank membranes. The polymer and plasticizer were dissolved in 10 mM phosphate buffer at pH 8.5. The pH of the solution was controlled using a FiveEasy pH meter (Mettler - Toledo) and adjusted using 1 M NaOH or 1 M HCl. A peptide solution with the desired concentration was prepared in a plastic bottle and used to achieve the target loading in the membrane. The required volume of the peptide solution was added immediately to the polymer solution before casting. The loaded membranes were fabricated on a Mathis SVLabcoater (Werner Mathis AG) to avoid loss of the peptide due to adsorption.

[0183] Most membranes were fabricated as a single matrix system to better characterize each matrix independently. As a test and proof of concept, a bilayer membrane: PVAGly_HPMCPVP_S2S was cast. For this, the basic procedure described previously was followed, but two different solutions containing their respective formulations were prepared and cast side by side.

[0184] For small-scale fabrication, such as samples for Fourier transform infrared spectroscopy (FTIR) analysis, a hydrophobic membrane was adhered to a flat surface. Then, the polymer-peptide solution was carefully placed on the hydrophobic material using a pipette and air-dried for 48 hours. This method produced small round discs of membrane material.

[0185] 1.4. Formulations

[0186] Different combinations of polymers and plasticizers were formulated in this study. All the tested formulations and the analyses performed on each prototype are summarized below:

[0187] Table 5 Tested formulations and analyses performed.

[0188]

[0189]

[0190]

[0191] Bioadh: Bioadhesion; Aggreg: Aggregation; Disso: Dissolution; Rel: Release

[0192] It is worth noting that only one formulation using Soluplus was prepared. In fact, after casting and drying, this membrane was almost inseparable from the casting surface (plastic or glass). Due to manufacturing difficulties and the limitation of the peptide on the casting substrate, Soluplus was not further considered.

[0193] 1.5. Bioadhesion

[0194] The adhesion of the polymer membranes was tested on porcine esophageal tissue, which is a recognized model mimicking the behavior of human oral tissue. Briefly, the freshly harvested esophagus was trimmed 1 - 2 cm from both ends. Then, the submucosa was carefully incised vertically and separated from the connective tissue by careful slicing. Then, the mucosa was rinsed in Krebs - Ringer buffer (KRB), cut into appropriate-sized pieces, and stored in KRB at 2 - 7 °C or frozen sandwiched between paraffin films for later use.

[0195] Bioadhesion was measured on a TA.XTplusC Texture Analyzer (Stable Micro Systems) calibrated at 500 g. The bioadhesive cells holding the tissue were immersed in artificial saliva (DPBS15 containing 0.75 g / L xanthan gum) slightly above the tissue level to ensure its moisturization. The tissue was always covered with 1 mL of artificial saliva to simulate the saliva accumulation effect in the oral cavity. The temperature was maintained at 37 °C under magnetic stirring on an IKA RCT standard (IKA Works) hot plate equipped with a thermal probe. The adhesion force was measured by lowering the polymer film (pre-cut into a circle with a diameter of 1.8 cm) attached to the probe using double-sided tape and applying a force of 100 g to the tissue within 30 s. The maximum adhesion force and the work of adhesion (area under the curve) were recorded at a contraction speed of 1 mm / s. Blanks were collected using only the metal probe, and the said data were subtracted from the measured values. Outliers were determined by the Grubb's test at a 0.05 confidence level and removed from the mean.

[0196] 1.6. Aggregation analysis by FTIR

[0197] Since P11-4 was designed to aggregate at physiological pH, a method for detecting aggregation in membranes was developed.

[0198] For self-assembling peptides (SAPs), such as P11-4, the assembled or aggregated state is decisive for their effect. Therefore, the peptide needs to remain in monomeric form in and upon release from the membrane. According to the existing literature on P11-4, its aggregation seems to be mainly driven by the formation of β-sheets. Therefore, the formation of this structure can be observed in the amide I band of FTIR, i.e., two characteristic peaks were identified in the aggregation spectra at 1684 cm−1 and 1617 cm−1, as shown by Davies et al. (2006. Supramolecular Chemistry 2006, 18, 435 - 443).

[0199] Therefore, spectra between 1000 and 3500 cm−1 (2 cm−1 step, 24 scans) were acquired in triplicate, along with spectra of blank formulations, using a UART Two (PerkinElmer) FT-IR device. The blank spectra were then subtracted from the corresponding formulation spectra using the SpectraGryph 1.2 software. Subtraction was performed using the coefficients provided by the software by default for the samples.

[0200] To evaluate whether this method was able to detect aggregation at low peptide concentrations, 10 mM phosphate buffer solutions containing 0.5% peptide at pH 5 or pH 8.5 were analyzed.

[0201] At low pH values, the peptide aggregated as expected, and the FT-IR method was able to detect a peak at 1684 cm-1, which is characteristic of this structure. On the other hand, for the high pH solution, no peak was detected, which is consistent with the monomeric state that should be observed at this pH value.

[0202] 1.7. Dissolution

[0203] Cut membrane patches with dimensions of 2 × 4.5 cm and place them between two glass microscope slides. Measure the thickness of three points of the patch using a Mitutoyo thickness gauge. Then use double-sided tape to stick the membrane to the bottom of a 100 mm plastic petri dish. Place the petri dish on an orbital shaker at 200 rpm in an oven at 37 °C and add 20 mL of artificial saliva (as described herein) to each petri dish.

[0204] Evaluate the dissolution of the membrane by observing it every 10 minutes to limit temperature changes in the oven. If in doubt, gently brush the double-sided tape with a spatula to check for the presence of a gel-like layer.

[0205] Since the low peptide loading target of this project was not expected to have a significant impact on the dissolution time, this experiment was conducted on blank formulations.

[0206] Each formulation was analyzed three times and the average dissolution time was determined.

[0207] 1.8. Peptide dosage

[0208] In this study, a plate-based intrinsic fluorescence quantification method was developed without using the provided HPLC method. The rationale behind this choice is that the former method is relatively simple to develop. There is no need to filter the samples, avoiding compatibility issues between the polymer and the column.

[0209] In a 96-well UV plate, place 50 μL of sample in each well. Excite the plate at 280 nm and measure the emission at 355 nm. For each formulation, calibration curves with concentrations of 1, 5, 10, 20, 35, and 50 μg / mL were plotted on the same plate. The calibration samples were prepared in artificial saliva containing excipients that mimic the matrix of the formulation. Each sample was measured three times.

[0210] 1.9. Release

[0211] Using membranes loaded with 0.5% peptide content, set up the release as described in the dissolution above.

[0212] At fixed time points (5 minutes, 15 minutes, 30 minutes, 45 minutes, 60 minutes, 90 minutes), take 300 μL of the release medium from each petri dish and replace it with fresh artificial saliva. Further assay the samples using the method described previously.

[0213] Each formulation was analyzed in triplicate. Results are expressed as the mean release at each time point relative to the final release at each replicate.

[0214] 2. Results and Discussion

[0215] Bioadhesion

[0216] Bioadhesion tests were conducted to (1) investigate the effects of plasticizers on film adhesion, (2) compare lead polymer candidates to commercially available whitening films, and (3) test whether peptides affect film adhesion.

[0217] from Figure 5 As can be seen, PVA and PVP show an increasing trend in both maximum peel force and work of adhesion with increasing plasticizer content. Plasticizers help increase chain free volume, promote hydration, and increase contact area with tissue and adhesion. This is demonstrated by the brittleness of PVP films without plasticizer, while slight incorporation significantly improves flexibility and adhesion.

[0218] However, increasing the plasticizer content did hinder the bioadhesion of the polymer, particularly in the placebo formulations of HPMC and PVA_Gly30. Excessive amounts caused the mixture to separate, coating the membrane in slippery plasticizer and hindering adhesion. This effect was most pronounced in formulations PVP_Gly15, PVP_Gly30, PVP-PEG30, and PVA_PEG30, and therefore was not tested. This leakage could potentially shorten the shelf life of the product and facilitate diffusion of the peptide from the formulation.

[0219] The bioadhesive properties are comparable to those of commercially available products.

[0220] After confirming the dissolution time of the lead formulation based on adhesion data, pure HPMC and PVA_Gly5 were selected to verify the effect of the peptide on membrane bioadhesion. Surprisingly, at the low peptide loadings tested, incorporation of P11-4 increased the overall adhesion properties of both tested formulations.

[0221] To further test whether PVP increases membrane adhesion when mixed with less adhesive polymers, a proof-of-concept membrane was fabricated using a 2:8 ratio of PVP to HPMC loaded with 0.5% P11-4. Surprisingly, the results showed an overall decrease in adhesion compared to the HPMC-loaded membrane. One possible explanation is demixing of the polymers during drying, which also explains the whitening of the membrane (not shown) due to the formation of newly formed domains. Including a plasticizer and carefully selecting the blend ratio can yield membranes with the desired properties; however, as shown, using membranes containing only one polymer is also a promising solution.

[0222] 2.2. Dissolution

[0223] Based on the bioadhesion results and considerations of other properties of the first formulated preparation (brittleness, plasticizer leakage...), four favorable preparations were selected for the dissolution test. These preparations were: PVA_pure, PVA_Gly5, PVP_Gly5, and HPMC_pure.

[0224] The dissolution times measured for these films and the tested film thicknesses (standard deviation) are as Figure 6 shown. The following table shows the dissolution times.

[0225] It can be seen from these data that the tested preparations showed different dissolution times, ranging from 15 to 50 minutes. Binary or ternary mixtures can be used to optimize the dissolution time and release performance.

[0226] In addition, the preparation containing HPMC had the longest dissolution time, followed by PVA and PVP. As expected, adding plasticizers such as glycerol to the preparation made the dissolution faster, as shown for PVA.

[0227] It is worth noting that the standard deviation of the dissolution time of PVA_Gly5 was narrower than that of PVA_pure. This improvement in reproducibility may have different explanations, such as a more reproducible thickness, or more uniform hydration due to the more hydrophilic glycerol.

[0228] 2.3. Release

[0229] After the dissolution test, two preparations, HPMC_pure and PVA_Gly5, were selected for the release study because they showed properties close to the target characteristics. In fact, PVP_pure seemed to have a very fast dissolution rate. In addition, since HPMC_pure showed a rather long dissolution time, PVA_Gly5 was selected instead of PVA_pure because its slightly higher dissolution rate and lower variability seemed more promising compared to the release curve of HPMC_pure. In addition, an additional preparation consisting of a mixture of PVP and HPMC was prepared to benefit from the extended dissolution time of HPMC_pure and the high bioadhesion of PVP. Therefore, the preparations tested by the release method were: HPMC_pure_Loaded, PVA_Gly5_Loaded, and HPMC_PVP_Loaded. The results are as Figure 7 shown.

[0230] Interestingly, the release curve of PVA_Gly5_Loaded was slower than those of the two HPMC-based preparations, i.e., faster than the release curves based on the HPMC matrix. This was surprising because, considering the dissolution times, this trend was the opposite.

[0231] In addition, it can be seen that although the thicknesses of the HPMC_pure_Loaded and HPMC_PVP_Loaded membranes are similar, the latter releases the peptide at a slower rate at shorter time points. This is unexpected because PVP_Gly5 showed the fastest dissolution time in the dissolution experiment. Therefore, it was thought that PVP would increase the peptide release rate of the HPMC-based membrane.

[0232] 2.4. Aggregation

[0233] Although the formulation was buffered at pH 8.5 before drying, during the drying step, due to water evaporation, an increase in the concentration of the self-assembling peptide and / or an increase in the salt concentration may have a negative impact and lead to self-assembly. Therefore, ATR-FTIR was used to evaluate the monomer state of the peptide in the dried membrane formulation. The spectra obtained from spectral acquisition and corresponding subtraction of the blank spectrum were analyzed (data not shown). In any of the tested formulations, the typical β-sheet band of the aggregated self-assembling peptide was not seen. Therefore, it is possible to provide membranes that are substantially or only composed of monomeric self-assembling peptides, such as monomeric P11-4. This is very important for therapeutic uses, such as the treatment and prevention of non-cavitated dental caries.

[0234] 2.5. Preparation of double matrix membranes

[0235] After demonstrating the feasibility of incorporating monomeric P11-4 into the polymer membrane, studies were conducted to investigate the preparation of double matrix membranes.

[0236] For example, two different dried membranes were simply fused together by depositing a concentrated polymer solution along the seam of the two membranes in a thin line. However, this was not very uniform, most likely due to the different water absorption rates of the two polymers. In fact, one of the sheets absorbed water much faster than the other, swelled and formed waves, which would hinder the proper fusion of the two sheets. The problem is less if the polymer matrices on both sides are the same or substantially the same (e.g., different only in the content of the plasticizer (and of course the active ingredient)).

[0237] In a second test, as described in Section 2.3, the two solutions were cast side by side and dried. As Figure 8 shown in B, this method produced a single membrane with the expected double matrix structure.

[0238] In the obtained side-by-side matrices, the interface between the two matrices is not completely straight, which may not only be impractical for the intended application to the gums and teeth, but also may mean variability in dosage (if the two matrices have different loads) and release profiles on both sides. In addition, the left side of the double matrix film composed of the PVA_Gly5 formulation appears less translucent than the pure PVA_Gly film on its left side. This indicates that some of the more opaque HPMC_PVP matrix on the right side has diffused and mixed during drying. This lateral transport may occur during the preparation of the "side-by-side film". Compared with the expectation of the "pure" matrix, the transfer may change the release profile, lead to inconsistent release and bioadhesion between batches, and cause bidirectional contamination of the opposite sides.

[0239] If two films are cast side by side simultaneously, a device that allows the casting of films with a straight interface in a reproducible manner can be used. For example, it includes a physical separation between the double film areas, which can be removed during the polymerization process. Faster drying methods may also be beneficial as this may limit the diffusion between the matrices.

[0240] Combining two already dried films can limit the problems of diffusion and non-straight interfaces.

[0241] The clamp will allow the films to be clamped flat while leaving a gap to add the polymer solution. A third dry film sprayed with an appropriate amount of water, or a third matrix with a low water content, can also be used as a substrate or support to bond the two matrices together. This will limit the absorption and diffusion of water between the matrices, while providing an opportunity to add an outer layer with other properties, such as a non-stick surface to protect the surrounding tissue or flavoring agents, etc.

[0242] 3. Conclusions and Outlook

[0243] Promising films containing active agents (such as self-assembling peptides) and capable of releasing monomeric self-assembling peptides within 30 to 45 minutes were successfully formulated for the first time. For example, oral films containing the P11-4 peptide were developed. ATR-FTIR was used to confirm their ability to keep P11-4 in an unaggregated state. In addition, these films were shown to have satisfactory bioadhesion (comparable to commercial products), disintegration time, and targeted release characteristics. The polymer matrices and excipients used are all materials approved for human use.

[0244] These results are very promising and demonstrate the feasibility of developing a film formulation for oral use.

Claims

1. A membrane (5) comprising a first surface (4), said first surface (4) comprising at least one first region (1) containing a first active agent and at least one second region (2) containing a second active agent different from said first active agent, Among them, Said at least one first region (1) is capable of contacting at least one tooth of a subject, and said at least one second region (2) is capable of contacting the subject's gingiva simultaneously.

2. The membrane (5) according to claim 1, wherein the membrane (5) is soluble after being administered in the oral cavity, wherein, Preferably, the membrane substantially dissolves after 1 - 240 minutes.

3. The membrane (5) according to any one of the preceding claims, wherein said first region (1) is adjacent to said second region (2) in said first surface (4).

4. The membrane (5) according to any one of the preceding claims, wherein the first region (1) is formed by a first sheet (10) and the second region (2) is formed by a second sheet (20), wherein, The surfaces (10, 20) of said first sheet and second sheet together form said first surface (4).

5. The membrane (5) according to claim 4, wherein said first sheet (10) or said second sheet (20) overlaps the corresponding other region by 10 - 100%.

6. The membrane (5) according to claim 4, wherein said first sheet (10) and said second sheet (20) are adjacent to each other.

7. The membrane (5) according to any one of claims 1 - 2 or 4 - 6, wherein said first sheet and said second sheet (10, 20) are separated by a spacer region (3), wherein said spacer region (3) is configured to inhibit the diffusion of at least one active agent from one region to another region, Optionally, wherein said first region (1), said second region (2) and, if present, said spacer region (3) are regions of the same sheet forming the membrane (5).

8. The membrane (5) according to any one of the preceding claims, wherein said first sheet (10) and said second sheet (20) forming said first surface (4) are arranged on a backing layer (30) to form a second surface (6) opposite to said first surface (4), Among them, Optionally, said backing layer (30) is configured to inhibit the diffusion of at least one active agent to the oral mucosa of the subject.

9. The membrane (5) according to claim 8, wherein, At least one selected from said backing layer (30), said spacer region (3), said first region (1) and said second region (2) is configured to release ions when applied to said at least one tooth and gingiva, Optionally, wherein the membrane comprises a battery (31), such as a bio - battery.

10. The membrane (5) according to any one of the preceding claims, wherein said membrane (5) or at least one of its sheets comprises a polymer material and / or a hydrogel as a matrix material, Wherein said matrix material optionally comprises a vinyl polymer, a polyacrylate, a PEEK (polyetheretherketone) polymer, carbomer, a natural polymer or a copolymer thereof, or pectin - coated liposomes.

11. The membrane (5) according to any one of the preceding claims, wherein said membrane (5) or at least one of its sheets comprises polyvinyl acetate (PVA), polyvinylpyrrolidone (PVP) and / or hydroxypropyl methylcellulose (HPMC), optionally PVA.

12. The membrane (5) according to any one of the preceding claims, wherein said membrane (5) or at least one of its sheets contains PVP.

13. The membrane (5) according to any one of the preceding claims, wherein the membrane (5) or at least one sheet thereof comprises HPMC.

14. The membrane (5) according to any one of the preceding claims, wherein the membrane (5) or at least one sheet thereof comprises a plasticizer, optionally glycerol or PEG-400, preferably glycerol.

15. The membrane according to claim 14, wherein the weight percentage of the plasticizer in each weight of the polymer and the plasticizer is 0-15%, preferably 0-5%.

16. The membrane (5) according to any one of the preceding claims, wherein the first active agent is selected from peroxides, reagents capable of generating peroxides, self-assembling peptides, CPP-ACP, mineral particles, and fluorides.

17. The membrane (5) according to any one of the preceding claims, wherein the second active agent is selected from anti-inflammatory agents, senescent cell scavengers, plant preparations, antibacterial agents, and self-assembling peptides.

18. The membrane (5) according to any one of the preceding claims, wherein at least the first region (1) comprises a self-assembling peptide, the self-assembling peptide comprising the consensus sequence SEQ ID NO:1, X1-X2-X1-X2-X1, wherein X1 is independently selected from glutamic acid, aspartic acid, glutamine, and ornithine, and X2 is independently selected from alanine, valine, isoleucine, leucine, methionine, phenylalanine, tyrosine, tryptophan, and glutamine, wherein the self-assembling peptide preferably has at least 80% sequence identity with SEQ ID NO:6 and most preferably comprises SEQ ID NO:

6.

19. The membrane according to claim 18, wherein the self-assembling peptide is in monomeric form.

20. The pharmaceutical membrane (5) according to any one of the preceding claims, optionally for treating at least one periodontal or dental disease or disorder selected from dental caries, mucositis, gingivitis, periodontitis, peri-implantitis, and tooth discoloration in a subject.

21. A method of treating a subject using the membrane (5) according to any one of claims 1 to 20, optionally a cosmetic method, comprising contacting at least one tooth of the subject with the first region (1) of the membrane and contacting at least one region of the subject's gingiva with the second region (2) of the membrane.

22. A method of preparing the membrane (5) according to any one of claims 1-20, optionally comprising three-dimensionally printing the membrane (5) in a personalized manner, taking into account the size and shape of the tooth, as well as the condition or disorder of the subject and its severity.

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