Self-assembling peptides in the prevention and treatment of cavitated carious lesions

Abstract

The present invention relates to the prevention and treatment of dental caries, particularly minimally or non-invasive dental caries treatment. It relates to compositions and kits comprising a self-assembling peptide, such as the self-assembling peptide P11-4 or P11-8, and a dental agent, i.e., a dental sealant, such as a glass ionomer cement-based sealant or a resin-based sealant, a fluoride varnish, a dental restorative material, or a bonding agent. The present invention also relates to compositions and kits comprising the self-assembling peptide and a dental agent for use in treating carious lesions, preferably cavitated caries lesions. The self-assembling peptide prevents secondary caries after the failure of a dental agent used to treat cavitated caries, particularly in the case of a failed interface between the dental agent and the tooth. The compositions and kits of the present invention may also be used for pulp capping.

Description

[Technical field]

[0001] The present invention is in the field of caries treatment, especially minimally or non-invasive dental caries treatment. It relates to compositions and kits comprising a self-assembling peptide, such as the self-assembling peptide P11-4 or P11-8, and a dental agent, i.e. a dental sealant, for example a cement-based sealant, such as a glass ionomer cement-based sealant, or a resin-based sealant, a fluoride varnish or bonding agent, or other dental restorative material. The present invention further relates to compositions and kits comprising a self-assembling peptide and a dental agent for use in treating carious lesions, preferably cavitated carious lesions. The carious lesions can be primary or secondary caries. Primary caries is not associated with a previous restoration. In contrast, secondary caries is defined as a lesion associated with a dental restoration or sealant. In the context of the present invention, the self-assembling peptide prevents secondary caries after failure of a dental agent used in treating cavitated caries, especially in the case of a failed interface between the restoration and the tooth with the dental agent. The compositions and kits of the present invention may also be used for pulp capping. [Background technology]

[0002] Tooth caries, also known as dental caries, is one of the most ubiquitous bacterial infections in the world. It is the breakdown of tooth material by bacterial metabolites, mainly acids produced when bacteria break down food debris or sugars on the tooth surface or in the biofilm. This leads to an imbalance between the demineralization and remineralization processes. The hard tooth structures, i.e. enamel, dentin and cementum, are damaged by ongoing demineralization, causing carious lesions and ultimately resulting in the appearance of carious cavities. The earliest sign of a new carious lesion is the appearance of chalky white spots on the tooth surface, the so-called white spot lesions (also called early carious lesions), i.e. subsurface lesions. As demineralization progresses, the mineralized surface of the lesion (partially) disintegrates and breaks down, revealing microcavities or cavities, i.e. holes in the tooth. This is called a (partially) cavitated carious lesion or a cavitated carious lesion.

[0003] Caries prevention approaches have been described that include topical fluoride therapy and the application of dental sealants, also known as pit and fissure sealants. The application of a sealant involves the introduction of the sealant into the pits and / or fissures of a tooth that is prone to caries. The main types of sealant materials currently on the market are resin-based sealants and glass ionomer cement-based sealants (Naaman et al., 2017, Dent J., 5, 34). The fluoride-releasing properties are considered one of the advantages of glass ionomer cements (Sidhu and Nicholson, 2016, J Funct Biomater., 7, 16), which thus combine the remineralizing effect of fluoride and the protective effect of sealants against pits and fissures.

[0004] To date, treatment of caries usually depends on whether the carious lesion is cavitated or non-cavitated. Non-invasive approaches have been proposed for non-cavitated carious lesions, i.e., subsurface lesions. For example, remineralization is attempted by application of topical fluoride.

[0005] Furthermore, enamel matrix derivatives or self-assembling peptides have been shown to be effective in remineralizing subsurface carious lesions (Ruan et al., 2013. Acta Biomater. 9(7):7289-97; Ruan et al., 2014. J Vis Exp. 10(89), doi 10.379151606; Schmidlin et al., 2016, J Appl Oral Sci. 24(1), 31-36; Alkilzy et al., 2018, Adv Dent Res. 29(1), 42-47; Brunton et al., 2013, Br Dent J, 215:E6; Kind et al., 2017, J Dent Res., 96(7), 790-797; Kirkham et al., 2007, J Dent Res., 86, 426-430). In WO2014 / 027012A1, a method is described for targeted delivery of self-assembling peptides to subsurface dental lesions with the aim of remineralizing the carious lesions. Alkilzy et al. (2018, supra) found an additional effect of application of P11-4, a self-assembling peptide, to early carious lesions in combination with fluoride, compared to application of fluoride alone.

[0006] Amelogenin-based biomineralization approaches for tooth repair have been described in the context of dentin hypersensitivity, tooth whitening or bleaching, and caries treatment in WO2017 / 123986A1, US2014 / 0186273A1, and US2017 / 0007737A1.

[0007] In WO2017 / 168183A1 biomimetic mineralized apatite structures based on elastin-like peptides are described for use as dental restorative materials, in particular for the reconstruction of enamel and for the treatment of dental diseases such as dental caries.

[0008] Furthermore, casein-based strategies have been developed for the remineralization of enamel and the prevention and / or treatment of caries or dental erosion (e.g. WO00 / 06108A1 and WO2010 / 042754A2). For example, casein phosphopeptide amorphous calcium phosphate complex (CPP-ACP) and CPP-stabilized amorphous calcium fluoride phosphate complex are described in WO02 / 094204A1 in combination with glass ionomer cement as remineralizing dental restorative materials. Some studies have also shown that caries progression slows or stops under sealants when applied to non-cavitated carious lesions (Griffin SOet al., 2008, J Dent Res., 87(2), 169-174).

[0009] When carious lesions are cavitated and holes appear in the teeth, the typical treatments applied so far are invasive, since remineralization at this stage is much more difficult than for non-cavitated carious lesions. The decayed material is usually removed, for example, using a dental handpiece ("drill"). Alternatively, lasers, dental spoons, or chemical-mechanical systems may be used to remove dental caries.

[0010] After removal of the carious enamel (i.e., drilling), the missing tooth structure requires dental restoration using dental restorative materials such as sealants, amalgam, dental composites, porcelain or gold. This traditional invasive treatment of cavitated caries is associated with several disadvantages.

[0011] First of all, during the process of removing the carious dental material, the natural tooth structure is severely damaged because healthy tooth material is also removed. Typically, the process of removing carious material is associated with pain and anxiety, which usually needs to be alleviated by application of local anesthetics. Furthermore, the removal of carious enamel affects the adjacent tooth structure.

[0012] Furthermore, restorations, especially with sealants or composites, often fail and eventually require redoing the filling, resulting in further loss of tooth structure. Failure may be caused by the presence or formation of a gap between the tooth and the restoration and / or sealant that is partially or completely lost. Such failures are often associated with secondary caries, also called recurrent caries or CARS (caries adjacent to restorations and sealants). The site of the filling serves as a vulnerable site for further caries (Burke and Lucarotti 2018. The ultimate guide to restoration longevity in England and Wales. Part 3: Glass ionomer restorations-time to next intervention and to extraction of the restored tooth. Br Dent J. 224(10):789-800; Nedeljkovic et al., 2015. Dental Materials 31(11):e247-e277). Summary of the Invention

[0013] There is therefore a need for an improved treatment of cavitated caries that overcomes at least some of the disadvantages of the prior art methods, in particular preventing the development of secondary caries. This problem is solved by the present invention, in particular the subject matter of the claims. Surprisingly, the inventors of the present invention have found that self-assembling peptides can be effectively used in combination with conventional dental agents, i.e. dental sealants, such as glass ionomer cements, fluoride varnishes or bonding agents for the advantageous treatment of cavitated carious lesions.

[0014] Thus, the present invention relates to a composition comprising (a) a self-assembling peptide, and (b) a dental agent selected from the group consisting of dental restorative materials such as dental sealants, fluoride varnishes and bonding agents, wherein preferably the dental agent is a dental sealant, e.g. a cement-based sealant, most preferably a glass ionomer-based sealant. The composition may be a pharmaceutical composition.

[0015] Preferably, in the compositions of the present invention, the self-assembling peptide and the dental agent are in a homogeneously mixed form. "Homogeneously mixed" means that the composition is a consistent mixture of its components. Thus, preferably, the compositions according to the present invention have essentially the same ratio of its components throughout any given sample.

[0016] The present invention further provides a kit comprising (a) a self-assembling peptide and (b) a dental agent, wherein the dental agent is a glass ionomer-based sealant. The kit may be a pharmaceutical kit.

[0017] The kit comprises the components of the composition according to the invention in separate form, for example in separate vials, flasks, syringes, applicators, or other containers. In the kit according to the invention, the self-assembling peptide may be included in a composition that may further comprise a solvent, such as water and / or a buffer. Alternatively, non-polar or polar organic solvents may be used. In the context of the present invention, unless otherwise stated, "a" is taken to mean "one or more". The kit may also comprise further components, for example two or more of the dental agents defined herein. For example, the kit may comprise, in addition to the self-assembling peptide, a complex and a binding agent suitable for binding the complex. Further desired components are, for example, etching agents and / or buffers that allow or demonstrate calcium, fluoride and / or phosphate.

[0018] The kit according to the present invention may comprise (a) a composition comprising a self-assembling peptide, and (b) at least one dental agent as defined herein, where the composition and the dental agent are in separate forms. Alternatively, the self-assembling peptide and a solvent such as water may be in separate forms, for example, separate compartments of an application device (e.g., commercially available from Curodont® Repair, credentis AG, Windisch, CH). The self-assembling peptide may be in a dried and buffered form, as disclosed in WO2014 / 027012.

[0019] A composition or kit that includes a self-assembling peptide means that the composition may contain a single type of self-assembling peptide, or two or more different types of self-assembling peptides, such as 3, 4, or 5. For example, as described in more detail below, the composition may contain two complementary peptides that self-assemble in combination.

[0020] As used herein, the term "comprises" encompasses the meaning of "consisting of."

[0021] The invention further provides a composition or kit comprising (a) a self-assembling peptide and (b) a dental agent as defined herein for use in treating a carious lesion, wherein the carious lesion is preferably a cavitated carious lesion, more preferably an active carious lesion such as an active cavitated carious lesion, and wherein preferably the composition or kit is a composition or kit according to the invention, such as a kit comprising a self-assembling peptide and a dental sealant.

[0022] The present invention also provides a composition or kit comprising (a) a self-assembling peptide, and (b) a dental agent selected from the group consisting of a dental sealant, a fluoride varnish, and a dental bonding agent, for use in preventing secondary caries following failure of a dental agent in treating a cavitated carious lesion. Preferably, the dental agent is a dental sealant, preferably a glass ionomer cement-based sealant.

[0023] In the context of the present invention, the term "carious lesion" (or "carious lesion") refers to the localized destruction of dental hard tissues by acidic by-products from bacterial fermentation of dietary carbohydrates. Dental caries can be defined as a microbial disease of the mineralized tissues of the teeth, characterized by demineralization of the inorganic parts and destruction of the organic material of the teeth. The term includes, for example, enamel caries, cement caries and dentin caries. In the context of the present invention, typically the lesion to be treated is a carious enamel lesion. Carious lesions may be subdivided into non-cavitated or subsurface carious lesions and cavitated carious lesions. The carious lesions to be treated in the present invention may be primary or secondary carious lesions.

[0024] A "cavitated carious lesion" refers to a carious lesion that has resulted in a tooth surface that is not intact, i.e., where there is at least partial destruction of the surface of the tooth structure in the area of ​​the carious lesion, e.g., a hole, e.g., localized destruction of the enamel.

[0025] There are indices used in dentistry to score the severity and activity of carious lesions.In the context of the present invention, preferably the clinical scoring system ICDAS (International Caries Detection and Assessment System), preferably the ICDAS-II system, is used.Information on the scoring and activity criteria of carious lesions according to ICDAS-II can be found, for example, in Dikmen (2015, J Istanbul Univ Fac Dent 49(3):63-72) or at www.icdas.org. In summary, the ICDAS-II stages describe the dental condition as follows: ICDAS-II stage 0: healthy; ICDAS-II stage 1: first visual changes in enamel; ICDAS-II stage 2: clear visual changes in enamel; ICDAS-II stage 3: localized enamel destruction (without clinical visual signs of dentin involvement); ICDAS-II stage 4: dark shadow at the bottom from dentin; ICDAS-II stage 5: clear cavities with visible dentin; ICDAS-II stage 6: extensive clear cavities with visible dentin. The ICDAS system is also used for Lesion Activity Assessment (LAA), typically this is based on a combination of the clinical appearance of the lesion, whether the lesion is in an area of ​​plaque stagnation, and knowledge of the tactile sensation as a ball-ended WHO probe is gently drawn across the tooth surface. Further information regarding the LAA may be found, for example, in Ekstrand et al. (2007, Oper Dent. 32(3):225-235) or Nyvad et al., 2018. Caries Res. 52(5):397-405).

[0026] In a preferred embodiment, the composition or kit for use according to the invention is for use in the treatment of a carious lesion at ICDAS-II stage 3, ICDAS-II stage 4, ICDAS-II stage 5 or ICDAS-II stage 6, preferably at ICDAS-II stages 3 to 6, such as ICDAS-II stages 3 to 5, even more preferably at ICDAS-II stages 3 to 4. Preferably, the carious lesion is an active carious lesion.

[0027] The detection and evaluation of carious lesions may be carried out by conventional caries detection and evaluation methods. For example, visual inspection assisted by a probe such as a ball-end probe, or radiography or a combination thereof is widely used. In the context of the present invention, a "cavitated carious lesion" may be a carious lesion detectable by a ball-end probe, such as a probe ball-end with a sphere showing a diameter of 0.5 mm (e.g., WHO probe). Information on the diagnosis and evaluation of dental carious lesions can be found, for example, in Braga et al. (2010, Dent Clin N Am 54:479-493).

[0028] Thus, for example, the invention provides a composition comprising (a) a self-assembling peptide, and (b) a dental agent as defined herein for use in treating, e.g., a carious lesion of ICDAS-II stage 1, 2, 3, 4, or 5, preferably a cavitated carious lesion, more preferably a carious lesion of ICDAS-II stage 3, 4, or 5, even more preferably a carious lesion of ICDAS-II stage 3 or 4. Further, for example, the invention provides a kit comprising (a) a self-assembling peptide, and (b) a glass ionomer cement-based dental sealant for use in treating a cavitated carious lesion, i.e. a carious lesion of ICDAS-II stage 3, 4, 5, or 6, more preferably a carious lesion of ICDAS-II stage 3, 4, or 5, even more preferably a carious lesion of ICDAS-II stage 3 or 4, wherein preferably the carious lesion is active.

[0029] Preventing secondary caries after failure of dental agent in treating hollow carious lesion means reducing the incidence of secondary caries. Typically, secondary caries forms under or at the edge of restoration. Dental agent failure can be the presence or formation of gaps between dental agents used to repair hollow carious lesion (partial failure). It can also be dental agent or restoration partially or completely worn or falling off (complete failure). In one embodiment, the treatment described herein not only reduces the incidence of secondary caries after failure of dental agent, but also increases the time to partial or complete failure of dental agent, because secondary caries at the edge of restoration, for example, can also cause such failure. However, this is not necessary, and it is sufficient that secondary caries is prevented after partial or complete failure of dental agent.

[0030] Typically, restorations classically made with dental agents as defined herein fail in a relatively short time, which is often associated in the art with secondary caries. For example, fluoride varnishes typically last for up to 3 days under conditions present in the oral cavity, including abrasion by chewing. In protected areas, the varnish can also be maintained longer. Typically, sealants, such as glass ionomer-based cement sealants, fail slowly over a period of 2-10 years. Typically, bonding agents are used to bond restorations, such as composites or crowns, to the tooth surface. Typically, failure of bonding agents and restorations are related. The time to failure of dental agents depends on several factors in addition to the material, namely, the patient (e.g., a history of extensive dental procedures is associated with a high failure rate), hygiene, the technical proficiency of the dentist, location, etc.

[0031] In the context of the present invention, the inventors have found that the combination of dental agents and self-assembling peptides prevents secondary caries, because upon failure of the dental agent, for example, the formation of a gap between the restoration with the dental agent or the dental agent worn / shedded, or the restoration bonded to the tooth with the dental agent, the matrix formed by the self-assembling peptides that allows remineralization protects the tooth from the onset of secondary caries. The remineralization of the matrix formed by the self-assembling peptides may be induced already under the cover of the dental material, depending on the availability of ions (e.g., from the dental material and / or saliva), when the tooth (i.e., the cavity) comes into contact with the self-assembling peptide. In any case, upon failure of the dental material, for example, when a gap is formed between the material and the tooth, the matrix formed by the self-assembling peptides can be remineralized and protect the tooth from secondary caries. The self-assembling peptides can also organize around the edges of the restoration with the dental material and protect these sites, which are also highly vulnerable to the onset of secondary caries.

[0032] When the self-assembling peptide and dental agent are mixed, the dental agent may further function as a delayed release reservoir for the self-assembling peptide.

[0033] Pretreatment with self-assembling peptides has been used to increase the bond strength of demineralized dentin (Barbosa-Martins et al., 2018. J Mech Behav Biomed Mater 81:214-221). It is known in the art that bonding of restorations, especially composites, to dentin is problematic because, compared to enamel, collagen is present as an organic component, and the water content of dentin is much higher and the concentration of ions is much lower than in enamel. Barbosa-Martins et al. tested whether pretreatment with self-assembling peptides would increase the bonding of two adhesive systems, Adper Single Bond (SB) and Clearfil SE Bond (CSE). Both agents are resin-based composites. An advantage was found for bonding with SB but not CSE. This may be due to the fact that SB requires etching with phosphoric acid. Phosphoric acid is thought to promote the organization of the self-assembling peptides. In contrast, CSE is a self-etching system and the pH drop is much less. Because there are significant differences between the dentin and enamel surfaces, this publication can only draw conclusions about dentin lesions and not about enamel lesions. Enamel lesions are preferred throughout the present invention. The publication also gives no indication as to whether the results apply to dental agents other than composites. In one embodiment of the present invention, the dental agent is not a resin-based composite. Indeed, as described herein, other dental agents, such as glass ionomer cement-based dental sealants, have advantageous properties in combination with self-assembling peptides.

[0034] Dental agents as defined herein are well known to those skilled in the art, and typically they are commercially available. Dental agents include dental sealants, dental varnishes, such as fluoride varnishes, and dental bonding agents. Thus, the present invention provides a composition or kit comprising a dental agent (in the case of the kit, a glass ionomer cement-based sealant) selected from the group consisting of (a) a self-assembling peptide, and (b) a dental restorative material, such as a dental sealant, a dental varnish, and a dental bonding agent, as described above, for use in treating carious lesions, preferably ICDAS-II stages 1 to 6, more preferably for use in treating hollowed carious lesions, such as ICDAS-II stages 3, 4, or 5, and in particular for use in preventing secondary caries.

[0035] The dental agent used in the present invention may comprise an acidic polymer selected from the group comprising acrylates and methacrylates, ionomers, diomers, Ormocer® and any other suitable polymers and / or monomeric forms thereof. Preferably, the dental agent comprises a component that can be polymerized, for example at or before being filled into a hollowed carious lesion. Thus, the components of the dental agent may initially be in the form of a monomer, which upon polymerization of said monomeric components may form a restoration (particularly in the case of a sealant) or a protective layer (particularly in the case of a varnish) or may be used to bond a restoration (particularly in the case of a bonding agent). The dental agent may be a material suitable for curing with visible light, an autopolymerizing material, or a material suitable for curing with UV light, preferably a material suitable for curing with visible light.

[0036] In one embodiment of the present invention, the dental agent is a dental varnish, such as a fluoride varnish. Fluoride varnishes are well known to those skilled in the art and are commercially available. In general, conventional fluoride varnishes contain fluoride as a salt or silane preparation at high concentrations in adhesive solutions, such as fast-drying alcohol and resin-based solutions. The concentration, form, and method of dispensing of the fluoride vary from manufacturer to manufacturer. Typical concentrations are, for example, 2.5% to 5% sodium fluoride or about 1% difluorosilane in a polyurethane, polyacrylic, colophonium, or shellac base. The varnish may contain CPP-ACP, such as MI Varnish (GC, Japan). Further suitable fluoride varnishes include, for example, Duropha® Suspension (Gaba Schweiz AG), Profluoride Varnish and Bifluorid 12 (Voco, Cuxhaven, Germany), Fluor Protector (Ivoclar / Vivadent, Amherst NY), Duraflor (PharmaScience, Montreal, Canada), CavityShield (OMNII Oral Pharmaceuticals, West Palm Beach, FL, USA) and Carex (Voss, Norway). Information regarding fluoride varnishes can be found, for example, in Chu and Lo (Gen Dent. Jul-Aug 2006;54(4):247-253).

[0037] In a preferred embodiment, the dental agent is a dental sealant. Dental sealants, also known as pit and fissure sealants, are well known to those skilled in the art and are widely used and commercially available in the field of dentistry. Information regarding dental sealants can be found, for example, in Naaman et al. (Dent J. 2017, 5, 34). In the context of the present invention, the dental sealant may be selected from the group including glass ionomer cement-based sealants, resin-based sealants, and modified ceramic materials (e.g., Ormocer®). Glass ionomer cement-based sealants include glass ionomer cements, resin-modified glass ionomer cements. Resin-based sealants include polyacid-modified composite resins (also called "compomers") and resin-based composites (also called composite resins), such as flowable composite resins.

[0038] Preferably, the dental sealant is a cement-based sealant, in particular a glass ionomer cement-based sealant. Thus, in one embodiment, the present invention provides a composition or kit comprising (a) a self-assembling peptide, and (b) a glass ionomer cement-based sealant, such as a resin-modified glass ionomer cement, a glass ionomer cement, or a glass ionomer type material. The composition or kit may be for use in treating carious lesions, for example, ICDAS-II stages 1 to 6. Typically, the sealant is used in treating non-cavitated carious lesions, and for this purpose may be used in combination with a self-assembling peptide, such as P11-4, either in a mixed form or in a kit form, where the peptide may be for initial administration, followed by administration of the sealant.

[0039] The present invention further opens the possibility to treat e.g. cavitated carious lesions of ICDAS-II stages 3-5, especially 3-4, with a glass ionomer cement-based sealant in combination with a self-assembling peptide such as P11-4. The advantage is that in combination with a self-assembling peptide, secondary caries is prevented (or the incidence of secondary caries is reduced).

[0040] Glass ionomer cements belong to a class of materials known as acid-base cements and are widely used and commercially available in the field of dentistry. Information on glass ionomer cements can be found, for example, in Sidhu and Nicholson (J Funct Biomater. 2016, 7, 16). The terms "glass ionomer cement" and "glass ionomer" are used interchangeably. Another term used for dental glass ionomer cements is "glass polyalkenoate cements". The main components of glass ionomer cements are polymeric water-soluble acids, basic (ion-leachable) glasses, and water. The polymers commonly used in glass ionomer cements are polyalkenoic acids. The glasses used in glass ionomer cements are basic and therefore capable of reacting with acids to form salts, for example aluminosilicate glasses with the addition of fluorides and phosphates are often used. Typically, commercial glasses for glass ionomer cements are based on calcium or strontium compounds and contain fluoride. Typically, glass ionomer cements harden within 2-3 minutes of mixing through an acid-base reaction and adhere to the tooth surface through chemical bonds formed by ionic bonds between the carboxylic acid groups of the polyacid molecules and the calcium ions on the tooth surface.

[0041] In the context of the present invention, "Glass Carbomer®" (GCP Dental, Netherlands) is also included in the term "glass ionomer cement-based sealants". It is a new commercial material of the glass ionomer type that contains substances that are not often included in glass ionomer formulations, such as silicone oils containing polydimethylsiloxane. Similarly, "Activa" is a rubberized glass ionomer cement-based sealant and is also included in the term "glass ionomer cement-based sealants".

[0042] Resin-modified glass ionomer cements may also be used as dental sealants in the present invention. These materials contain the same essential components as conventional glass ionomers (basic glass ionomer, water, polyacid), but also include a monomer component and an associated initiator system. Typically, the monomer is 2-hydroxyethyl methacrylate (HEMA) and the initiator can be camphorquinone or other suitable photoinitiators.

[0043] In another embodiment, resin-based sealants, i.e. resin-based dental sealants, are used as the dental sealant of the present invention. Resin-based sealants are well known in the dental field and are typically based on an oligomeric matrix containing components such as bisphenol A-glycidyl methacrylate (bisGMA), urethane dimethacrylate (UDMA), or other dimethacrylate monomers (such as TEGMA or HDDMA), or semicrystalline polyceram (PEX). They may be classified into four generations determined by the polymerization method. The first generation was polymerized by the action of ultraviolet light. The second generation was an autopolymerizing resin-based sealant or a chemically cured sealant. In this case, a tertiary amine (activator) is added to one component and mixed with another component. The reaction between these two components generates free radicals that start the polymerization of the resin sealant material. The third generation includes visible light polymerized resin-based sealants, and thus typically visible light in the wavelength range of about 470 nm activates a photoinitiator (e.g., as described herein) present in the sealant material. The fourth generation of resin-based sealants is fluoride resin-based sealants, which are the product of adding fluoride-releasing particles into conventional resin-based sealant materials. Resin-based sealants can be filled or unfilled, i.e., they can contain filler materials, for example, as described herein for resin-based composite materials. Typically, unfilled resin-based sealant materials exhibit lower viscosity. Suitable commercially available resin-based sealants include Clinpro® (3M-ESPE, St. Paul, MN, USA), Delton® FS (Dentsply-De Trey, Konstanz, Germany), Estiseal® F (Heraerus-Kulzer, Hanau, Germany), and Guardian Seal® (Kerr, Orange, CA, USA).Thus, in one embodiment, the invention provides a composition or kit comprising (a) a self-assembling peptide, and (b) a resin-based sealant, e.g., for use in treating carious lesions. Typically, the resin-based sealant is used in the treatment of non-cavitated carious lesions, and for this purpose may be used in combination with a self-assembling peptide, such as P11-4.

[0044] The present invention further opens the possibility to treat cavitated carious lesions, e.g. ICDAS-II stages 3-5, especially 3-4, with resin-based sealants in combination with self-assembling peptides such as P11-4. The advantage is that in combination with self-assembling peptides, secondary caries is prevented (or the incidence of secondary caries is reduced).

[0045] As used herein, "resin-based sealant" also includes resin-based composites, preferably flowable resin-based composites (also known as flowable resin composites). Thus, resin-based composites, preferably flowable resin-based composites, may be used as dental sealants or preventative resin restorations in the present invention. The terms "resin-based composites" and "composite resins" are used interchangeably herein. Resin-based composites are a type of synthetic resin that is widely used in dentistry as restorative materials, dental adhesives, and sealants. Conventional resin-based composites include a resin-based oligomer matrix and a filling material, for example, based on glass or ceramic materials. For example, bisphenol A-glycidyl methacrylate (bisGMA), urethane dimethacrylate (UDMA) or other dimethacrylate monomers (such as TEGMA or HDDMA), or semi-crystalline polyceram (PEX) are used as the resin-based oligomer matrix. Typically, glass fillers are made of crystalline silica, silicon dioxide, lithium / barium-aluminum glass, or borosilicate glass containing zinc / strontium / lithium. Ceramic fillers include zirconia silica and zirconium oxide. Typically, composite resins also contain initiator systems such as photoinitiators, e.g., camphorquinone, phenylpropanedione (PPD), and lucrin (TPO). These initiator systems initiate the polymerization reaction of the resin when irradiated with blue light. There are also chemically cured resin composite systems (also known as "autopolymerizing" resin-based sealants), which are usually provided as two paste systems (substrate and catalyst) and begin to cure when the substrate and catalyst are mixed together. Various additives may be added to control the reaction rate or physical properties of the dental composite resin, for example, dimethylglyxomine, which may be added to achieve a particular flowability. Dental composites exhibit the ability to adhere to tooth structure through micromechanical bonding that is increased by acid etching. With the additional application of dental bonding agents, very high bond strengths to tooth structure can be achieved.

[0046] Resin-based composites may be classified into "flowable" and "filled" categories depending on their handling properties. Typically, flowable resin composites exhibit a filler content of less than 60% by volume, such as about 35-55% by volume. Typically, filled resin composites exhibit a higher filler content, for example, greater than 60% by volume. In general, flowable resin composites exhibit lower viscosity and therefore better wetting properties and ease of handling compared to filled resin-based composites.

[0047] Suitable commercially available composite resins include, for example, Metafil (Sun Medical, Moriyama, Japan), Heliomolar® (Ivoclar-Vivadent, Liechtenstein), Solitaire® (Heraeaus Kulzer, Hanau, Germany), Arabesk (Vocvo, Cuxhaven, Germany), and Charisma® (Heraeaus Kulzer, Hanau, Germany). Examples of suitable commercially available flowable resin composites are Admira® Flow (Voco, Cuxhaven, Germany), Filtek® Supreme (3M-ESPE, St. Paul, MN, USA), FlowLine (Heraeus-Kulzer, Hanau, Germany), Grandio® Flow (Voco, Cuxhaven, Germany), Point-4® Flowable (Kerr, Orange, CA, USA), Premise® Flowable (Kerr, Orange, CA, USA), Revolution® Formula 2 (Kerr, Orange, CA, SA), and X-Flow® (Dentsply-De Trey, Konstanz, Germany). Thus, in one embodiment, the present invention provides a composition or kit comprising: (a) a self-assembling peptide; and (b) a resin-based composite material, preferably a flowable resin-based composite material, preferably for use in the treatment of a carious lesion, more preferably for use in the treatment of a cavitated carious lesion, such as an ICDAS-II stage 3, 4 or 5 carious lesion, as described herein.

[0048] As used herein, resin-based sealants also include polyacid-modified composite resins (also called "compomers"). Thus, the dental sealant used in the present invention may be a polyacid-modified composite resin (also called "compomer"). Compomers resemble conventional composite resins in that the curing reaction is typically an addition polymerization, does not contain water, and the majority of the components are identical. Typically, these are bulky macromonomers such as bisglycidyl ether dimethacrylate (bisGMA) or its derivatives and / or urethane dimethacrylates, blended with a viscosity-reducing diluent such as triethylene glycol dimethacrylate (TEGDMA). These polymer systems are filled with non-reactive inorganic powders such as quartz or silicate glass, e.g., SrAlFSiO4. These powders are usually coated with silanes to promote bonding between the filler and the matrix in the cured material. Typically, compomers contain additional monomers that are different from conventional composites, including acid functional groups. For example, 2-hydroxyethyl methacrylate and diesters of butanetetracarboxylic acid (TGB). The curing reaction is usually initiated by light, but in some systems the curing occurs as a result of mixing two pastes, each of which contains a component of a free radical initiator system. Examples of suitable commercially available compomers include Ana Compomer (Nordiska Dental, Sweden), Dyract® (Dentsply, Konstanz, Germany), Compoglass® (Ivoclar-Vivadent, Liechtenstein), F2000 Compomer (3M-ESPE, St. Paul's, MN, USA), Freedom (SDI, Bayswater, Victoria, Australia), Hytac® (3M-ESPE, Seefeld, Germany), MagicFil (Zenith, Englewood, NJ, USA), Twinky Star (Voco, Cuxhaven, Germany).Thus, in one embodiment, the present invention provides a composition or kit comprising (a) a self-assembling peptide, and (b) a polyacid-modified composite resin, preferably for use in the treatment of a carious lesion, more preferably for use in the treatment of a cavitated carious lesion, such as an ICDAS-II stage 3, 4 or 5 carious lesion, as described herein.

[0049] In another embodiment, the dental agent used in the present invention is a dental bonding agent, also called "dentin bonding agent". Dental bonding agents are well known in the field of dentistry and are typically resinous materials traditionally used to firmly bond dental composite filling materials to both dentin and enamel. Often dental bonding agents are methacrylates with volatile carriers and solvents, such as acetone or ethanol / water. As adhesives, typically BisGMA or TEGMA are used. Application of dental agents usually requires the use of an etchant, such as phosphoric acid, citric acid / calcium chloride, oxalic acid / aluminum nitrate, and a primer, such as NTG-GMMA / BPDM, HEMA / GPDM or 4META. Dental bonding agent systems are available that combine all the necessary components in one composition. Others provide the components separately. Suitable commercially available dental bonding agents include 3M® Scotchbond® Universal Adhesive (3M ESPE, St. Paul, MN, USA), ACE® ALL-BOND (BISCO Dental Products, Schaumburg, IL, USA), Admira Bond (VOCO, Cuxhaven, Germany), BeautyBond 7 thExamples of suitable dental bonding agents include 1-generation adhesive (Shofu Dental Corporation, Ratingen, Germany), CLEARFIL SE Bond 2 (Kuraray America Inc., Houston, TX, USA), Futurabond U (VOCO, Cuxhaven, Germany), G-Premio BOND® (GC America Inc., Alisp, IL, USA), and Prelude® (Zest Dental Solutions, Carlsbad, CA, USA). Thus, in one embodiment, the present invention provides a composition or kit comprising (a) a self-assembling peptide and (b) a dental bonding agent, preferably for use in treating a carious lesion, more preferably for use in treating a cavitated carious lesion, such as an ICDAS-II stage 3, 4 or 5 carious lesion, as described herein.

[0050] In the present application, two or more types of dental agents may be used.For example, the kit according to the present application may include a dental sealant based on glass ionomer cement and a fluoride varnish.Therefore, for example, when using such a kit to treat carious lesion, the self-assembling peptide may be applied first to the tooth surface that is optionally cleaned and / or treated, for example, the carious lesion surface (either before the fluoride varnish or in one composition with the fluoride varnish), and the glass ionomer cement may be applied after the application of the fluoride varnish.

[0051] In one embodiment of the present invention, no fluoride varnish is applied.

[0052] In the context of the present invention, the dental agent, for example a glass ionomer cement, a calcium silicate-based cement, a polyacid-modified composite resin, or a resin-based sealant, is preferably capable of releasing fluoride. In a particularly preferred embodiment, the dental agent is capable of releasing calcium and fluoride. For example, a glass ionomer cement is based on calcium aluminofluorosilicate glass powder or a strontium-based glass and an acid, typically polyacrylic acid, and therefore has both properties. The dental agent may also or alternatively be permeable to ions, for example from saliva, especially after it has been applied to the tooth. Thus, substances present in saliva, such as calcium or phosphate, can penetrate the dental agent. These two features improve the remineralization of the tooth, especially the cavity surface, already before the dental agent fails, thus improving protection.

[0053] There are dental agents such as dental sealants with a basic pH, and there are dental sealants such as dental sealants with a neutral or acidic pH. The pH of the dental agent can be determined before the material hardens, especially in an aqueous environment. For example, many cements, such as Portland cement, are basic. In the context of the present invention, both basic or neutral or acidic dental agents may be used, but this may affect the type of self-assembling peptide used, as will be described in more detail below.

[0054] Portland cement is an example of a calcium silicate-based cement that may be used in the context of the present invention. It is preferably used for dentin restorations close to the dental pulp because of its relatively long setting time. For example, mineral trioxide aggregates (MTA), such as fast-setting mineral trioxide aggregates, such as Biodentine (Septodont, Saint Maur des Fosses, FR); Watson et al., 2014. Dental Mater. 30(1):50-61), which sets within about 12 minutes, can be used.

[0055] The self-assembling peptides used in the present invention are peptides that can form three-dimensional scaffolds, thereby promoting tissue regeneration. They may assemble in one dimension to form higher-order assemblies, such as beta sheets and tape-like assemblies. Three-dimensional supramolecular structures of self-assembling proteins with affinity for / to calcium phosphate can be formed.

[0056] In the context of the present invention, self-assembling peptides may be capable of self-assembling by themselves, as is the case for example for peptides P11-4, P11-8, P11-2, P11-5 below, or alternatively, they may self-assemble in combination of two self-assembling peptides, as is the case for example for peptides P11-13 / P11-14 and P11-28 / P11-29, P11-30 / P11-31 below.

[0057] In the context of the present invention, the self-assembling peptides taught in WO2004 / 007532A1, US10 / 521,628, US12 / 729,046, US13 / 551,878, US14 / 062,768 or WO2014 / 027012A1 are preferred, all of which are fully incorporated herein by reference.In particular, the self-assembling peptides having a net charge of +2 or -2 at pH 7.5 may be used in monomeric or assembled form.

[0058] Preferably, the self-assembling peptide used in the present invention comprises 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 peptide used in the present invention comprises the consensus sequence SEQ ID NO: 1, X1-X2-X1-X2-X1, where X1 is independently selected from the group consisting of glutamic acid, aspartic acid, glutamine, glutamine and ornithine, and X2 is independently selected from the group consisting of alanine, valine, isoleucine, leucine, methionine, phenylalanine, tyrosine, tryptophan and glutamine, or an amino acid sequence having at least 80% identity thereto.Independently selected means, for example, that X1 at position 1, 3 or 5 of the above sequence can be different from each other. Of course, they can also be the same.

[0059] Preferably, the self-assembling peptides used in the present invention comprise SEQ ID NO:2, X1-X2-X1-X2-X1, where X1 is independently selected from the group consisting of glutamic acid and ornithine, and X2 is independently selected from the group consisting of tryptophan and phenylalanine.

[0060] In a further embodiment, the self-assembling peptide used in the invention may comprise SEQ ID NO:3, X3-F-X1-W-X1-F-X1, where X1 is independently selected from the group consisting of glutamic acid and ornithine, and X3 is selected from the group consisting of arginine, glutamic acid and ornithine, where preferably X3 is arginine.

[0061] Furthermore, the self-assembling peptide used in the present invention may comprise SEQ ID NO: 4, or preferably consists of X4-X4-X3-F-X1-W-X1-F-X1-X4-X4, where X1 is independently selected from the group consisting of glutamic acid and ornithine, X3 is independently selected from the group consisting of arginine, glutamic acid and ornithine, and X4 is independently selected from the group consisting of glutamine, glutamic acid, serine, threonine and ornithine. Preferably, X3 is arginine. Independently, preferably, X4 is glutamine.

[0062] The self-assembling peptides of the invention may comprise SEQ ID NO:5, or preferably consist of QQRF-X1-W-X1-F-X1-QQ, where X1 is independently selected from the group consisting of glutamic acid and ornithine.

[0063] Preferably, the self-assembling peptides used in the present invention comprise or consist of a sequence selected from the consensus sequences listed in Table 1 below.

[0064] Most preferably, the peptides comprise or consist of the specific peptides listed in Table 2. Of course, self-assembling peptides that assemble in combination with another self-assembling peptide, for example as disclosed herein, may be formulated in a single kit or composition.

[0065] Peptides of SEQ ID NO: 6, 9, 11, 12, 16 or 17 are particularly advantageous, for example because they can be used in relatively low concentrations, are highly compatible with cells and have a beneficial charge distribution.

[0066] Preferably, the self-assembling peptide comprises or consists of the sequence of SEQ ID NO: 6. The peptide consisting of the sequence of SEQ ID NO: 6 is also called P11-4 and is preferred throughout the present invention. In another preferred embodiment, the self-assembling peptide comprises or consists of the sequence of SEQ ID NO: 9 (P11-8).

[0067] The kit or composition of the invention may also comprise at least one self-assembling peptide having at least 45% sequence identity to a peptide consisting of SEQ ID NO: 6. Preferably, the peptide has or is at least 54%, at least 63%, at least 72%, at least 81% or at least 90% sequence identity to a peptide consisting of SEQ ID NO: 6. For example, a peptide of the invention is 11 amino acids in length.

[0068] The self-assembling peptides may be modified peptides containing an Ac-N-terminus and / or an NH2-C-terminus, preferably both, or unmodified peptides. The termini of all self-assembling peptides of SEQ ID NO: 1 are preferably blocked to enhance stability, since the unblocked forms tend to initiate deamination reactions. In particular, peptides of SEQ ID NO: 6, 9, 11, 12, 16 and 17 may contain an Ac-N-terminus and an NH2-C-terminus. SEQ ID NO: 18-29 correspond to modified peptides of the invention. [Table 1] [Table 2]

[0069] Preferably, the self-assembling peptides do not have restriction sites for the endopeptidase of interest. They also do not need to contain special recognition motifs for the cell.

[0070] In one embodiment, in the kit or composition according to the invention, the self-assembling peptide is mainly in monomeric form, for example, at least 70%, at least 80%, or at least 90% of the self-assembling peptide is present in a monomeric state. For this purpose, if the peptide assembles at a pH of 7.5 or less, the pH of the composition is preferably 0.1 to 0.5 pH units higher than the pH at which the peptide begins to undergo self-assembly (e.g., pH 7.5 for P11-4), or more than 0.5 pH units higher than the pH. To avoid rapid aggregation, the pH may be buffered at that pH. It may be beneficial if aggregation and hydrogel formation begins immediately after application to the carious cavity. Thus, the pH may be 0.1 to 0.5 pH units higher than the pH at which the peptide begins to undergo self-assembly without buffering. In one embodiment, the composition may comprise dry peptide, for example as obtainable according to WO2014 / 027012.

[0071] In another embodiment, a kit or composition according to the invention comprises a self-assembling peptide in a predominantly organized form, e.g., at least 70%, at least 80%, or at least 90% organized form, or essentially organized form, and a buffer (at a pH that stabilizes the organized form). Typically, the assembled self-assembling peptide forms a hydrogel.

[0072] The choice of protein assembly state depends on several factors. The predominantly monomeric form of the self-assembling peptide may improve the diffusion of the monomer into demineralized enamel and be particularly advantageous for smaller cavitation lesions, such as partially cavitation lesions or lesions with microcavities (e.g., ICDAS1-3). The predominantly assembled form may result in a more rapid action of the peptide and thus improve the function, especially for larger cavities (e.g., ICDAS4, 5, or 6). When the self-assembling peptide and dental agent are mixed, typically the pH and ionic strength required for the dental agent to harden are decisive. For compositions that include both dental agents and self-assembling peptides, either predominantly monomeric self-assembling peptides or predominantly polymeric self-assembling peptides may be used at the respective pH.

[0073] The combination of complementary self-assembling peptides, such as P11-4 and P11-8, provides significantly faster assembly times and, because they are attracted to each other, results in faster application and stability.

[0074] For example, the present invention provides a composition or kit comprising (a) a self-assembling peptide comprising a consensus sequence according to SEQ ID NO:1-5, such as a consensus sequence according to SEQ ID NO:1, preferably a consensus sequence according to SEQ ID NO:2, more preferably a consensus sequence according to SEQ ID NO:3, even more preferably a consensus sequence according to SEQ ID NO:4, and most preferably a consensus sequence according to SEQ ID NO:5, as described above, for use in treating a cavitated caries lesion, such as a caries lesion at ICDAS-II stage 4 or 5. In particular, it may be used for the prevention of secondary caries.

[0075] In a preferred embodiment, the present invention provides a composition or kit comprising (a) a self-assembling peptide comprising, preferably consisting of, a sequence selected from sequences according to SEQ ID NO: 6-17 and 30-33 or a sequence having at least 80%, more preferably at least 90% identity thereto, preferably a sequence according to SEQ ID NO: 6 or a sequence having at least 80%, more preferably at least 90% identity thereto, and (b) a dental agent such as a dental sealant, preferably a glass ionomer cement-based sealant, as described above, preferably for use in the treatment of carious lesions, both early and advanced carious lesions, more preferably for use in the treatment of cavitated carious lesions, such as carious lesions of ICDAS-II stages 3-5. P-11 is most preferred for use in the context of the present invention. In particular, it may be used for the prevention of secondary caries.

[0076] In certain preferred embodiments, the present invention provides a composition or kit comprising (a) a self-assembling peptide comprising, preferably consisting of, a sequence according to SEQ ID NO: 6, and (b) a glass ionomer cement-based sealant, as described above, preferably for use in the treatment of carious lesions, more preferably for use in the treatment of carious lesions of ICDAS-II stages 3 to 5, such as 4 or 5. In particular, it may be used for the prevention of secondary caries.

[0077] In the context of the present invention, the self-assembling peptide used may undergo self-assembly at low pH, particularly at a pH of less than 7.5, for example, P11-4. Typically, carious lesions have an acidic pH, ensuring the assembly of self-assembling peptides within the lesion. The preferred peptide that undergoes self-assembly at low pH, particularly at a pH of less than 7.5, is P11-4.

[0078] The term self-assembling peptide (SAP) self-assembles at a pH of less than 7.5 means that it can self-assemble at a pH of less than 7.5. Ionic strength also influences the assembly state of the selected self-assembling peptide. Preferably, the self-assembling peptide used in the present invention can self-assemble at a pH of less than 7.5 and at least physiological ionic strength. The self-assembling peptides that can self-assemble at a pH of less than 7.5 and at least physiological ionic strength can begin to undergo self-assembly at said pH, as for example the preferred peptide P11-4, but it is not necessary. They can also be in a self-assembly state at higher or lower pH.

[0079] Those skilled in the art know how to determine and measure the ionic strength of a solution. Ionic strength I is generally calculated according to the formula

number

[0080] If desired, the mechanical properties can be influenced by the concentration of the self-assembling peptide as well as the types of molecules and ions present in the composition. Compositions containing self-assembling peptides used in the present invention may include a biologically suitable buffer, such as, for example, NaCl, and optionally Tris.

[0081] In one embodiment, the assembly state of the self-assembling peptides may also be controlled by ionic strength, for example, assembly may be induced by combination with a buffer of high ionic strength (e.g., greater than 0.15 mol / L) that may contain calcium, phosphate and / or fluoride ions.

[0082] The pH in the lesion (or between the surface of the cavitated carious lesion and the dental agent) may be influenced by the pH of the dental agent. Most dental agents, such as chitosan-arginine-amide, have a neutral or acidic pH.

[0083] Alternatively, in the context of the present invention, the self-assembling peptide used may undergo self-assembly at high pH, ​​particularly at a pH of at least 7.5.In this case, typically, dental agent has a pH of at least 7.5, such as Portland cement or any calcium hydroxide-based material.The preferred peptide that undergoes self-assembly at high pH, ​​particularly at a pH of at least 7.5, is P11-8.

[0084] An aspect of the present invention is a composition or kit according to the present invention for use in the treatment of cavitated carious lesions as described above, preferably cavitated carious lesions of enamel, in particular for use in preventing or reducing the incidence of secondary caries after failure of dental agents. Thus, primary carious lesions may be treated, the incidence of secondary caries being reduced by the treatment of the present invention. Typically, the treatment of carious lesions in the context of the present invention is carried out by a dentist.

[0085] The treatment may include (a) cleaning the tooth surface, (b) optionally drying the tooth surface, (c) optionally etching the tooth surface, (d) administering a composition or component of the kit to the carious lesion, and (e) optionally curing, where steps (b) and (c), if present, may be performed in any order.

[0086] Typically, step (a) precedes steps (b) to (e) of the treatment procedure. Steps (b) to (d) are more flexible in the order of application, and step (e) usually follows steps (b) to (d). Thus, for example, etching (if applied) may be performed before or after drying, or during step (d), etc., and typically the order and application of steps depends on the type of dental agent used in the present invention, and corresponds to the order and application of steps typically taken for said dental agent when said dental agent is used without self-assembling peptide. For example, when a resin-based sealant is used as the dental agent, it is advantageous to etch the tooth surface before application of the resin-based sealant. Resin-based sealants are more sensitive to moisture than glass ionomer cements. Therefore, in the case of application of a resin-based sealant, it is advantageous to dry the tooth surface. Preferably, in this case, the tooth is dies and etched. Dentists know the treatment procedures that can be recommended for different dental agents.

[0087] Step (a), "cleaning the tooth surface" means that at least the surface of the carious lesion is cleaned. That is, substances such as food particles, dental deposits, plaque or dental film are removed from the tooth surface, at least from the surface of the carious lesion, by a suitable method. Such cleaning methods may include treatment with a conventional toothbrush, treatment with a specific dental brush, for example, a brush for removing plaque, treatment with a dental scaler and cullet, application of a mouthwash, for example, using an antibacterial mouthwash, etc. Preferably, cleaning includes removal of plaque. Cleaning may further include partial or complete removal of carious tooth material, for example, by drilling, air abrasion, or laser ablation, by application of a spoon excavator, or by chemical-mechanical removal (for example, using the Carisolv® system). In the context of the present invention, removal of carious tooth material is generally not necessary, and preferably drilling is not performed.

[0088] Nevertheless, in the case of carious lesions of more advanced caries stages, such as carious lesions of ICDAS-II stages 3, 4, 5, and 6, it may be advantageous to at least partially remove the decayed tooth material. If removal of carious tooth material is performed, it is preferred that the carious tooth material is removed by a "gentle" method, such as using a spoon excavator or chemical-mechanical removal method, rather than drilling. The inventors have found that in the context of enamel lesions without dentin involvement (preferred herein), it is not necessary to completely remove the carious material. It is particularly preferred that the process of removing the carious material does not remove intact tooth material, which is typically the case when drilling is used as the removal technique. In the case of lesions with dentin lesions, preferably the carious material is removed.

[0089] As mentioned above, the application of step (b), i.e. drying the tooth surface, may be advantageous for the application of certain dental agents, such as resin-based sealants, dental bonding agents, or resin-based composites. Drying may be performed before etching (if etching is performed), after etching, or in the context of step (d), for example between the application of the self-assembling peptide and the application of the dental agent. Typically, drying the tooth surface is performed using an air flow.

[0090] Furthermore, step (c), i.e. etching the tooth surface, may be advantageous for the application of certain dental agents, such as resin-based sealants, dental bonding agents, or resin-based composites. Etching is a common technique traditionally used in dental sealing procedures. "Etching" refers to creating microscopic porosity on the tooth surface into which dental agents, such as self-assembling peptides and / or dental sealant materials, can flow, thereby increasing retention, increasing surface area, and improving the strength of the bond between the dental agent and the tooth surface. Etching agents that may be used in the context of the present invention include maleic acid, EDTA, citric acid, tartaric acid, phosphoric acid, nitric acid, and polyacrylic acid. A particularly preferred etching agent is phosphoric acid, which is also often referred to as a conditioning agent. Typically, the etching agent is applied to the cleaned and optionally dried tooth surface for between 10 and 60 seconds, for example between 15 and 30 seconds. During etching, the tooth surface is preferably rinsed, for example with water. Depending on the dental agent to be applied, the tooth surface is dried, for example in the case of a resin-based sealant, after an optional rinsing step prior to application of the dental agent. In one embodiment, etching is performed after application of the self-assembling peptide and prior to application of the dental agent.

[0091] Step (d) comprises administering the composition or components of the kit to the carious lesion, preferably the hollowed carious lesion. This means that the composition or components of the kit are administered to the carious lesion by suitable techniques known to dentists, for example by filling. When the kit of the present invention is used to treat a carious lesion, preferably step (d) comprises (i) administering the self-assembling peptide or a composition comprising the self-assembling peptide (included in the kit) to the carious lesion, and then (ii) administering the dental agent to the carious lesion. Typically, "application of the dental agent" refers to application as professionally and conventionally performed by a dentist, for example according to the manufacturer's instructions. The self-assembling peptide may be administered, for example, as a solution in a suitable buffer. The self-assembling peptide may be administered to the carious lesion by any suitable applicator, for example a syringe, a sponge, etc. For example, the application used in Curodont® Repair may be used. Preferably, the kit also includes an applicator.

[0092] In one embodiment, step (d) comprises administering a mixture of the self-assembling peptide and the dental agent to the cavitated carious lesion. This mixture may be a composition according to the invention or a component of a kit of the invention that is mixed before or during administration. The mixture may be administered as is conventionally practiced in the dental field for the respective dental agent, for example according to the manufacturer's instructions.

[0093] The desired application of step (e), i.e., hardening, depends on the dental agent used. Most dental agents require some type of hardening. The term "hardening" refers to a process in which a chemical reaction (such as polymerization) or physical action (such as evaporation) occurs, resulting in a harder, stronger, or more stable connection (such as a bond) or substance (such as a hardened resin). Typical hardening for dental materials is chemical hardening, hardening by application of light, thermal hardening, or a combination thereof.

[0094] Thus, for example, the present invention provides a composition or kit according to the invention for use in the treatment of a carious lesion, preferably a cavitated carious lesion such as a carious lesion at ICDAS-II stage 3, 4 or 5, as described above, wherein the treatment of the carious lesion comprises the steps of: (i) cleaning the tooth surface including removing plaque, in particular cleaning the surface of the carious lesion, optionally including partial or complete removal of carious material, where preferably no drilling is performed, (ii) administering to the carious lesion a composition comprising a self-assembling peptide as described above, such as a self-assembling peptide comprising a consensus sequence according to SEQ ID NOs: 1-5 (e.g., P11-4), (iii) administering to the carious lesion a dental agent, such as a glass ionomer cement-based sealant or a fluoride varnish, and (iv) hardening the fluoride varnish, for example by drying, or hardening the glass ionomer cement-based sealant, for example by chemical hardening or a combination of chemical hardening and hardening with visible light. The order of steps may be from (i) to (iv).

[0095] Another example of a composition or kit for use according to the invention is, as described above, a composition or kit according to the invention for use in the treatment of carious lesions, preferably cavitated carious lesions such as carious lesions at ICDAS II stages 3, 4 or 5, wherein the treatment of the carious lesion comprises the following steps, preferably according to the manufacturer's instructions: (i) cleaning the tooth surface including removing plaque, in particular cleaning the surface of the carious lesion, optionally with partial or complete removal of carious material, where preferably no perforation is performed, cleaning; (ii) optionally removing the carious lesion from the tooth surface, preferably according to the manufacturer's instructions; (iii) applying an etching agent, such as phosphoric acid, to the surface of the carious lesion for a suitable period of time, thereby etching the surface of the tooth having the carious lesion; (iv) removing the etching agent, for example by rinsing; (v) administering to the carious lesion a composition comprising a self-assembling peptide as described above, such as a self-assembling peptide comprising a consensus sequence according to SEQ ID NOs: 1-5 (e.g., P11-4); (vi) drying the surface of the tooth having the carious lesion; (vii) administering a resin-based sealant; and (viii) curing the resin-based sealant by a suitable curing method.

[0096] The choice of dental agent in the context of the present invention may be influenced by the type of carious lesion to be treated. For example, the present invention advantageously allows the use of dental sealants for cavitated carious lesions of any size, or occlusal carious lesions in tooth pits and / or cracks.

[0097] Furthermore, the present invention provides a method for treating a hollowed carious lesion, comprising administering to said carious lesion a component of the kit or a composition of the present invention.For example, the method may comprise the above steps (a) to (e) as described above for the composition and kit for use in treating a carious lesion, namely (a) cleaning the tooth surface, (b) optionally drying the tooth surface, (c) optionally etching or conditioning the tooth surface, (d) administering to the carious lesion a composition or component of the kit, and (e) optionally curing.

[0098] The present invention further solves additional technical problems. In particular, during excavation of a carious lesion, the dental pulp may be exposed or nearly exposed, increasing the risk of pulpitis or pulp shrinkage, since the dental pulp reacts against the dental materials (sealants, bonding agents, fillers, resins, etc...). Surprisingly, the inventors have found that the kit or composition of the present invention can be used by dentists to prevent the dental pulp from deteriorating when a dental restoration approaches the dental pulp. The present invention therefore provides a composition or kit according to the present invention for use in pulp capping. The dental material may be a dental sealant or bonding agent, or a dental agent as mentioned above, such as a filling agent. It may be a glass ionomer cement-based sealant selected from the group comprising glass ionomer cements and resin-modified glass ionomer cements, or a resin-based sealant selected from the group consisting of polyacid-modified composite resins and resin-based composites. In this context, resin-based composites are the preferred dental materials.

[0099] For example, where the pulp is exposed or nearly exposed (to an extent that would increase the risk of pulp or pulpitis shrinkage, as can be appreciated by those skilled in the art), e.g., in the context of cleaning, particularly excavating, a carious lesion, a solution of a self-assembling peptide (e.g., P11-4 or a mixture of P11-4 and P11-8, preferably P11-4) may be applied to protect the pulp and improve remineralization, and then a dental material, e.g., a dental sealant, is applied.

[0100] Alternatively, the self-assembling peptide (e.g., P11-4 or a mixture of P11-4 and P11-8, preferably P11-4) and the dental material may be mixed prior to application, which is most relevant if the dental material is also biocompatible with the dental pulp (e.g., MTA).

[0101] The composition or kit of the present invention may be used for pulp capping.

[0102] The pulp capping may be direct or indirect pulp capping. The remineralization of the thin layer of softened dentin remaining for indirect pulp capping, which can be improved by the self-assembling peptide used in the context of the present invention, is particularly advantageous. Glass ionomers or resin-modified glass ionomers can also be used in the context of the pulp capping of the present invention, especially for indirect pulp capping. If necessary, a different layer of dental sealant can be applied on top of the pulp capping.

[0103] One source of calcium ions for remineralization may be the dental pulp, if the self-assembling peptide is placed on top of the dental pulp and protects it from shrinkage due to the cytotoxicity of the dental filling material. In the case of pulp capping, the ions for remineralization originate from the dental pulp (or via the blood supply of the dental pulp) and remineralization can begin immediately after placement (without / before the filling fails or the formation of microgaps or microleaks). Of course, additional ions, including fluoride, may originate from the dental material. Therefore, fluoride-releasing dental materials, such as glass ionomer cements, are also advantageous in this context.

[0104] The present invention is illustrated, but not limited, by the following examples. All references cited herein are incorporated in their entirety. [Brief description of the drawings]

[0105] Figure legend Figure 1.1 Treatment of a partially cavitated carious lesion (ICDAS class 3). A: Cross section of a partially cavitated tooth; partial cavitation on the left. B: Application of a mainly monomeric self-assembling peptide (e.g., P11-4, fragment) solution to the tooth surface after cleaning of the tooth, here without drilling. The self-assembling peptide starts to diffuse into the cavity. C: Self-assembling peptides, such as P11-4, assemble in the cavities due to the low pH and generate a matrix. D: The tooth surface is dry, leaving organized P11-4 ribbons and fibers within the lesion. E: After the solution is absorbed into the cavity, a sealant such as glass ionomer cement (GIC) is applied, allowing diffusion of ions (calcium and phosphate) from the saliva and the glass ionomer cement into the lesion. F: Calcification (hexagon) within the lesion protects the failed sealant, resulting in a calcified structure underneath the lesion (protected by the sealant, e.g., GIC). After failure of the GIC, a fully regenerated tooth surface and structure remains.

[0106] Figure 1.2 A: Cross section of a partially cavitated tooth; partial cavitation on the left. B: Excavated carious lesion C: Application of monomeric P11-4 (piece) simultaneously or sequentially with the filler. D: Extracted tooth with filling (gray) and fibril P11-4 (long pieces) at the interface between the tooth and filling E: Mineralization (hexagonal) in the microscopic gap between the filling and the tooth. Calcium phosphate is supplied either from the filling (in the case of glass ionomer cement), saliva, or pulp (via dentinal fluid).

[0107] Figure 2 Treatment of cavitated carious lesions (ICDAS class 4) with a composition of the invention in a caries model as described in Example 2. In the right and left columns, two representative examples are shown. A, B: Perforation of artificial tooth C, D: Application of 1 drop of P11-4, 10 mg / mL, pH 8 E, F: Application of glass ionomer cement (Aqual Ionofill Plus, VOCO)

[0108] Figure 3 Treatment of cavitated carious lesions (ICDAS class 4) with a kit of the invention in a caries model as described in Example 3. In the right and left columns, two representative examples are shown. A, B: Perforation of the artificial cavity. C, D: Application of glass ionomer cement (Aqual Ionofill Plus, VOCO) mixed with one drop of P11-4, 10 mg / mL, pH 8.

[0109] Figure 4. Release profiles of model substances Congo Red and P11-4 from varnish, glass ionomer cement, and Paro-Amin Fluor Gelee. Experiments were performed as specified in Examples 6 and 7. Release was measured as described and specified in arbitrary units (AU). Figure 4A shows the release of Congo Red from varnish Duraphat (sample A1) and glass ionomer cement Aqua Ionofil Plus (sample B1), and Figure 4B shows the release of Congo Red from paro Amin Fluor Gelee (sample 2). Figure 4C shows the release of monomer P11-4 from varnish Duraphat (sample AP) and glass ionomer cement Aqua Ionofil Plus (sample BP).

[0110] Working Example Example 1 A partially cavitated lesion (ICDAS class 3) is cleaned. It is either excavated (Figure 1.2) or not (Figure 1.1). After cleaning, optionally including excavation, a self-assembling peptide (e.g., P11-4) is applied to the cleaned tooth surface, especially the carious tooth. The self-assembling peptide starts to diffuse into the cavity, where it organizes due to the low pH and generates a matrix. After the solution is absorbed into the dentin or enamel (10 seconds to 5 minutes, e.g., depending on the surface, more rapid absorption in dentin than in enamel), a sealant such as a glass ionomer cement is applied, which transfers ions (Ca and phosphate) from the saliva or glass ionomer into the lesion body. This allows the initiation of mineralization in the matrix protected by the sealant. After the sealant falls off the tooth, typically after several months or years, a completely regenerated tooth surface and structure remains, free of secondary caries.

[0111] Example 2 Artificial ICDAS class 4 lesions, created by drilling in excised bovine incisors, were treated with the kit of the invention according to the manufacturer's instructions, specifically by application of one drop of P11-4, 10 mg / mL, pH 8, followed by glass ionomer cement (Aqual Ionofill Plus, VOCO). Briefly, polyacrylic acid conditioner was applied for 10 seconds, rinsed, lightly dried, the glass ionomer was applied, and the material was allowed to mature before the finishing procedure. The main steps are shown in Figure 2. The glass ionomer cement was cured to produce a hard restoration indistinguishable from the restoration applied without combination with P11-4 (data not shown). No differences could be determined up to one week after filing.

[0112] Example 3 An artificial ICDAS class 4 lesion created by drilling in a resected bovine incisor was treated with the composition of the present invention. A glass ionomer cement powder (Aqual Ionofill Plus, VOCO) was prepared according to the manufacturer's instructions. 1 mg of P11-4 was added per 100 mg (10 mg per ml of powder) and mixed with water. The powder was dissolved according to the manufacturer's recommendations. The solution was mixed for 30 seconds and applied to the tooth surface (as above) as recommended by the manufacturer. The main steps are shown in Figure 3.

[0113] Glass ionomer cement cured to produce hard restorations indistinguishable from restorations applied without combination with P11-4 (data not shown). Differences could not be determined until 1 week after filing.

[0114] Example 4 - Prevention of secondary caries Teeth with artificial ICDAS class 4 lesions generated by drilling in excised bovine incisors are treated with a dental sealant (e.g., a glass ionomer (e.g., Aqual Ionofill Plus, VOCO)) in the absence or presence of P11-4 according to three different protocols: 1) Apply dental sealant to the cleaned surface of the cavity according to the manufacturer's instructions. 2) Apply a drop of P11-4, 10 mg / mL, pH 8, to the cleaned surface of the cavity such that the surface of the cavity is covered. After the solution is essentially absorbed into the tooth surface, apply dental sealant to the cleaned surface of the cavity according to the manufacturer's instructions. 3) Prepare glass ionomer cement powder (Aqual Ionofill Plus, VOCO) according to manufacturer's instructions. Add 1 mg of P11-4 per 100 ml (10 mg per ml of powder) and mix with water. Dissolve powder according to manufacturer's instructions. Mix solution for 30 seconds as recommended by manufacturer and apply to tooth surface (as above).

[0115] The teeth were stored in distilled water for 24 hours. They were then thermocycled (500 cycles, 5-55°C), immersed in basic fuchsin, sectioned, and analyzed for dye penetration and remineralization using a stereomicroscope (e.g., as described in Hepdeniz et al., 2016. Eur J Dent. 10(2):163-169).

[0116] The formation of microgaps or microleakage and the shrinkage of the dental sealant are monitored over time and the remineralization at the site of the microgap is analyzed. The different groups are compared.

[0117] Example 5 - Pulp Capping - Dental pulp from blood supply and remineralization with calcium phosphate. A cavity exposing the pulp for dental caries is excavated, and the pulp is covered with P11-4 solution (e.g., 100 μl, 10 mg / mL, pH 8) before depositing (e.g., applying in layers and light curing, as appropriate) a dental material, such as a resin composite, according to the manufacturer's instructions.

[0118] The pulp remains asymptomatic (i.e., there is no pulpitis) and a follow-up radiograph one year later evaluates the pulp for shrinkage.

[0119] Example 6 - Release profile. As known to those skilled in the art, the release profile of a drug, i.e., fluoride or a self-assembling peptide, depends on the matrix of the material (Mousavinasab et al., 2009, Dent Res J (Isfahan) 6(2):75-81). In the following experiments, the release profile of a model substance mimicking the self-assembling peptide Congo Red was evaluated after incorporation into dental materials.

[0120] The materials were prepared and applied according to the instructions for use (IFU), which usually describe the use of 0.5-0.75 mL of the respective product. To mimic saliva, phosphate buffered saline (PBS) was used. PBS matches the osmolality and ionic concentration of the human body - it is isotonic. To mimic a constant saliva flux, the applied volume was placed in a solution equivalent to the 1-h saliva flux, which is approximately 20 mL. The diffusion of the material was evaluated at 500 nm using a photometer. [Table 3]

[0121] Release Profile a) Preparation of varnish (Duraphat) and paro Amin Fluor Gel According to the supplier IFU, it is recommended to add 0.5-1 mL of varnish to the carious site. The concentration of Congo Red was selected to lead to a clear signal in UV Vis. - Place 12mg of Congo Red in a 2mL Eppendorf tube -Add 2mL of varnish directly into the tube with a syringe - Mix Congo Red and varnish vigorously until content uniformity is reached. -Pick up the product with a syringe - 0.5-0.75 ml of product is deposited as a thin film on a glass slide as recommended in the IFU - Air dry according to IFU b) Preparation of Ionofil (glass ionomer restorative material) -12mg of Congo Red placed directly in the dental cup -Add 2 spoonfuls of Ionofil Plus as per IFU -Add 2 drops of water and mix according to IFU -Place a quantity of the product on a slide c) Incubation Prepare PBS solution with 1% EtOH for storage -Place the slide in a 50ml centrifuge tube -Add 20mL of PBS - Place it on the rotating wheel d) Measurements and Calculations - Decant 1mL of culture directly into the UV-Cell -500 nm, control / blank = PBS containing 1% EthOH -Subtract the control without Congo Red from the measurements

[0122] Results and Discussion A model substance that is easily incorporated into products regardless of material formulation. This is indicated by visual assessment of content uniformity as a uniform red / brown color dispersion is visible.

[0123] The model substances are also able to diffuse into the artificial saliva from all three fluoride-containing formulations. Depending on the formulation, the release profiles of the individual formulations differ.

[0124] The model material Congo red was released from the varnish Duraphat and glass ionomer cement (Figure 4A) as well as from the paro Amin Fluor Gel used for comparison (Figure 4B). The release from the varnish was more rapid than the release from the glass ionomer cement. The release from the glass ionomer cement was found to be constant and increased with time.

[0125] Release profile of Example 7-P11-4. Example 7 was carried out essentially as Example 6, but with the self-assembling peptide P11-4, unless otherwise stated. 12 mg of P11-4 (credentis AG) in monomeric form was added to either the varnish Duraphat (sample ID: AP) or the glass ionomer cement Aqua Ionofil Plus (sample ID BP) in the amounts and conditions described above. Incubation was carried out in a static manner at room temperature. Measurements were at 280 nm, and the control values ​​for buffer without peptide were subtracted from the measurements.

[0126] The results are shown in Figure 4C. Confirming previous data, the experiments show a surprisingly good diffusion of the peptide from the matrices tested, i.e. Duraphat (sample AP) or Aqua Ionofil Plus (BP), confirming the data obtained with model materials.

[0127] P11-4 is released slowly and substantially constantly from both the varnish and the glass ionomer cement. The amount of peptide released increases with time measured over several days. Release from the varnish is more rapid than from the glass ionomer cement. Thus, depending on the formulation, a slow release can be obtained by using a sealant such as a glass ionomer cement, or a "fast" release can be obtained with a varnish, e.g., a colophony-containing formulation.

[0128] A fast release, such as that seen with varnishes such as Duraphat, may have a beneficial effect on the performance of the peptide immediately after application, which may be particularly useful, for example, in the remineralisation of lesions covered by the varnish.

[0129] On the other hand, a longer release may be particularly advantageous for the prevention or treatment of secondary lesions, which may develop slowly over time under or at the edge of the sealant on the treated (filled) lesion. It is reasonable to conclude that the drug is released from the sealant, e.g., glass ionomer cement, over a longer period of time than from the varnish. This is appropriate because the average time on the tooth of the sealant, e.g., glass ionomer cement, is also longer than for the varnish. Furthermore, the self-assembling peptide released from the filling may remain in place for a long period of time.

Claims

1. a) a self-assembling peptide, where the self-assembling peptide is P11-4 consisting of the sequence of SEQ ID NO:6, and b) a dental agent selected from the group consisting of dental sealants, fluoride varnishes, and dental bonding agents; wherein preferably the composition is a pharmaceutical composition.

2. Dental drugs include: (i) a glass ionomer cement-based sealant selected from the group consisting of glass ionomer cements and resin-modified glass ionomer cements; and (ii) a resin-based sealant selected from the group consisting of polyacid-modified composite resins and resin-based composite materials.

10. The composition of claim 1, wherein the dental sealant is a glass ionomer cement based sealant.

3. The composition of claim 1 , wherein the dental agent is a fluoride varnish.

4. The composition of claim 1 , wherein the dental agent is a dental bonding agent.

5. In another format, a) a self-assembling peptide, where the self-assembling peptide is P11-4 consisting of the sequence of SEQ ID NO:6, and b) a dental agent which is a glass ionomer cement-based dental sealant selected from the group consisting of glass ionomer cements and resin-modified glass ionomer cements; wherein preferably the kit is a pharmaceutical kit.

6. A composition according to any one of claims 1 to 4 or a kit according to claim 5, wherein the dental agent is capable of releasing fluoride ions, preferably wherein the dental agent is also capable of releasing calcium ions.

7. The composition according to any one of claims 1 to 4 or 6 or the kit according to any one of claims 5 to 6, wherein the dental agent is permeable to ions.

8. The composition of any one of claims 1 to 4 or the kit of any one of claims 5 to 7, wherein the self-assembling peptide is predominantly in monomeric form.

9. The composition of any one of claims 1 to 4 or the kit of any one of claims 5 to 7, wherein the self-assembling peptide is in a predominantly assembled form.

Images