RU2828353C2 - Self-assembled peptides in prevention and treatment of cavities accompanied by formation of cavities - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: group of inventions relates to the field of agents for preventing and treating caries, particularly minimally invasive or non-invasive caries treatment, and concerns a composition and a kit, as well as use thereof. Disclosed composition and kit contain: a) a self-assembled peptide representing P11-4, having SEQ ID NO: 6, and b) a dental substance; wherein the kit contains the above components in separate form. Dental substance used in the composition is: dental sealant selected from the group of glass ionomer cements and polymer-modified glass ionomer cements; or fluorine lacquer containing ethyl alcohol as a solvent in combination with rosin or shellac base; set includes a dental sealant based on glass ionomer cement, selected from a group comprising glass ionomer cements and polymer-modified glass ionomer cements. Also disclosed is the use of said composition or kit in preventing secondary caries after unsuccessful use of a dental substance in treating a cavity-forming carious lesion, as well as in a protective coating of the pulp.

EFFECT: use of the self-assembled peptide P11-4 in the above composition and kit provides the prevention of secondary caries after the unsuccessful use of the dental substance used in the treatment of caries accompanied by cavity formation, especially in case of violation of boundary between restoration made of said dental substance, and tooth, and, besides, it is effective for protective coating of pulp.

15 cl, 7 dwg, 2 tbl, 7 ex

Description

The present invention relates to the field of caries treatment, in particular minimally invasive or non-invasive treatment of tooth decay. It relates to a composition and a kit comprising a self-assembling peptide, such as a self-assembling peptide P11-4 or P11-8, and a dental substance, namely a dental sealant, such as a cement-based sealant, such as a glass ionomer cement-based sealant or a polymer-based sealant, a fluoride varnish, or a bonding agent, or another restorative dental material. The invention further relates to a composition and a kit comprising a self-assembling peptide and a dental substance for use in the treatment of a carious lesion, preferably a carious lesion that occurs with cavity formation. Carious lesions can be primary or secondary caries. Primary caries is not associated with a previous restoration. Conversely, secondary caries is defined as a lesion associated with dental restorations or sealants. In the context of the invention, the self-assembling peptide prevents secondary caries after the destruction of a dental substance used in the treatment of caries accompanied by cavity formation, especially in the case of a disruption of the interaction between the restoration made with the dental substance and the tooth. The composition and kit according to the present invention can also be used for a protective pulp coating.

Tooth decay, also known as dental caries, is one of the most common bacterial infections in the world. It is the breakdown of dental substance due to bacterial metabolites, mainly acids produced by bacteria when they break down food debris or sugar on the surface or in the biofilm of the teeth. This leads to an imbalance between the processes of demineralization and remineralization. The hard structures of the teeth, i.e. enamel, dentin and cementum, are damaged by continuous demineralization, leading to carious lesions and, ultimately, to the formation of cavities. The earliest sign of a new carious lesion is the appearance of a chalky white spot on the surface of the tooth, the so-called white spot lesion (also called incipient carious lesion), i.e. a subsurface lesion. As demineralization progresses, the mineralized surface of the lesion is (partially) destroyed, and a microcavity or cavity, a hole in the tooth, is formed. This is called (partially) cavity forming caries or cavity forming caries.

Approaches to prevent caries have been described that include topical fluoride treatment and the use of dental sealants, also known as pit and fissure sealants. Sealant application involves the insertion of sealants into the pits and/or fissures of caries-prone teeth. 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 property is considered as one of the advantages of glass ionomer cements (Sidhu and Nicholson, 2016, J Funct Biomater, 7, 16), which thus combine the remineralizing action of fluoride and the protective action of pit and fissure sealants.

Today, caries treatment usually depends on whether or not the lesion is cavity-forming. Noninvasive approaches have been proposed for non-carious lesions that develop a cavity, i.e., subsurface lesions. For example, attempts are being made to remineralize the lesion by applying fluoride to the lesion.

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). Patent application WO 2014/027012 A1 describes a method for targeted delivery of a self-assembling peptide to a subsurface dental lesion for the purpose of remineralization of carious lesions (Alkilzy et al., 2018, see above) found an additional effect of the application of P11-4, a self-assembling peptide, on incipient carious lesions, in combination with fluoride, compared to fluoride alone.

An approach for tooth restoration based on amelogenin-based biomineralization has been described in the context of dentin hypersensitivity, tooth whitening or bleaching, and in the treatment of caries in patent applications WO 2017/123986 A1, 2014/0186273 A1 and 2017/0007737 A1.

In patent application WO 2017/168183 A1, a biosimilar mineralized apatite structure based on elastin-like peptides is described for use as a dental restorative material, in particular for the restoration of enamel and for use in the treatment of dental diseases, such as dental caries.

In addition, casein-based strategies have been developed for the remineralization of enamel and the prevention and/or treatment of dental caries or erosion (e.g., patent applications WO 00/06108 A1 and WO 2010/042754 A2). Casein phosphopeptide amorphous calcium phosphate complexes (CPP-ACP) and CPP-stabilized amorphous calcium fluoride phosphate complexes are described in combination with glass ionomer cement as remineralizing dental restorative materials, e.g., in patent application WO 02/094204 A1. Some studies have also shown that caries progression is slowed or stopped under sealants when applied to non-cavitating carious lesions (Griffin S.O. et al., 2008, J Dent Res., 87 (2), 169-174).

Once a carious lesion has developed a cavity and a hole has formed in the tooth, the general treatment used today is invasive, since remineralization at this stage is much more difficult than for carious lesions that do not form a cavity. The destroyed tissue is usually removed using, for example, a dental handpiece ("drill"). Alternatively, a laser, an excavator or a chemomechanical system can be used to remove dental caries.

After removal (i.e. excavation) of the enamel damaged by caries, the lost tooth tissue requires dental restoration using dental restorative materials such as sealants, amalgam, dental composites, ceramics or gold. This traditional invasive treatment for cavity-forming caries has several disadvantages.

Firstly, the natural tooth tissue is seriously damaged because healthy tooth tissue is also removed during the process of removing the damaged tooth tissue. The process of removing carious tissue is usually associated with pain and anxiety, which usually must be relieved by the use of local anesthetics. In addition, due to the removal of carious enamel, adjacent tooth tissue is affected.

In addition, restorations, particularly those with sealants or composites, frequently fail and the filling eventually has to be redone, resulting in further tooth loss. Failure may be due to the presence or development of gaps between the tooth and the restoration and/or the sealant partially or completely falling out. Such failure is often associated with secondary caries (also called recurrent caries or CARS). The filling site is vulnerable to further damage (Burke and Lucarotti 2018. Comprehensive guide to the longevity of restorations in England and Wales. Part 3: Glass ionomer restorations - time to next intervention and extraction of the restored tooth. Br Dent J. 224 (10): 789-800; Nedeljkovic et al. 2015. Dental Materials 31 (11): 247-277).

There is therefore a need for an improved treatment of cavity-forming caries that overcomes at least some of the aforementioned 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 invention according to the claims. The inventors of the present invention have surprisingly found that self-assembling peptides can be effectively used in combination with traditional dental substances, namely, a dental sealant, such as glass ionomer cement, fluoride varnish or a bonding agent, for the preferential treatment of cavity-forming carious lesions.

The present invention thus relates to a composition comprising (a) a self-assembling peptide and (b) a dental substance selected from the group consisting of a dental restorative material, such as a dental sealant, a fluoride varnish and a binder, wherein preferably said dental substance is a dental sealant, such as a cement-based sealant, most preferably a glass ionomer-based sealant. The composition may be a pharmaceutical composition.

Preferably, in the composition of the invention, the self-assembling peptide and the dental substance are in a homogeneously mixed form. "Homogeneously mixed" means that said composition is a mixture of these components. Thus, the composition of the present invention preferably has substantially the same proportions of its components in any given sample.

Furthermore, the present invention provides a kit comprising (a) a self-assembling peptide and (b) a dental substance, wherein said dental substance is a glass ionomer-based sealant. Said kit may be a pharmaceutical kit.

Said kit comprises the components of the composition of the present invention in separate form, for example, in separate jars, vials, syringes, applicators or other containers. In the kit of the present invention, the self-assembling peptide may be contained in a composition, which may further comprise a solvent, for example, water and/or a buffer. Alternatively, non-polar or polar organic solvents may be used. In the context of the invention, unless otherwise indicated, the word in the singular is interpreted as meaning "one or more". The kit may also comprise additional components, for example, more than one of the dental substances defined herein. For example, the kit may comprise, in addition to the self-assembling peptide, a composite and a binder suitable for binding to the composite. Additional optional components are, for example, an etching agent and/or a buffer capable of providing calcium, fluoride and/or phosphate.

The kit of the present invention may comprise (a) a composition comprising a self-assembling peptide and (b) at least one dental substance as defined herein, wherein said composition and said dental substance are in separate form. Alternatively, said self-assembling peptide and said solvent, such as water, may be in separate form, such as in separate compartments of an application device (such as the Curodont® Repair marketed by credentis AG, Windisch, CH). The self-assembling peptide may be in dried, buffered form, as described in patent application WO 2014/027012.

"A composition or kit comprising a self-assembling peptide" means that the composition may comprise one type of self-assembling peptides or two or more, such as three, four or five, etc., different types of self-assembling peptides. For example, as described in detail below, the composition may comprise two complementary peptides that self-assemble when combined.

The term "comprising" as used herein includes the meaning "consisting of."

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

The invention also provides a composition or kit comprising (a) a self-assembling peptide and (b) a dental substance selected from the group consisting of a dental sealant, a fluoride varnish and a dental bonding agent, for use in the prevention of secondary caries after unsuccessful treatment with a dental substance in the treatment of a carious lesion occurring with cavity formation. Preferably, the dental substance is a dental sealant, preferably a sealant based on glass ionomer cement.

In the context of the present invention, the term "caries" (or "carious lesion") refers to the localized destruction of the hard dental tissues by acidic by-products of bacterial fermentation of dietary carbohydrates. Caries can be defined as a microbial disease of the calcified dental tissues, characterized by demineralization of the inorganic part and destruction of the organic substances of the tooth. The term includes, for example, enamel caries, cementum caries and dentin caries. In the context of the invention, the lesions to be treated are typically enamel caries. Carious lesions can be divided into non-cavity or subsurface carious lesions and cavity-forming carious lesions. The carious lesions to be treated according to the invention can be primary or secondary carious lesions.

The term "cavitating carious lesion" refers to a carious lesion that results in the tooth surface no longer being intact, i.e., there is at least partial destruction of the tooth surface tissue in the area of the carious lesion, such as a hole, such as localized destruction of the enamel.

There are indices used in dentistry for scoring the severity and activity of carious lesions. In the context of the present invention, the clinical scoring system ICDAS (International Caries Detection and Assessment System), preferably the ICDAS-II system, is preferably used. Information on the scoring and activity criterion 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 general, the ICDAS-II stages describe the dental status as follows: ICDAS-II stage 0: intact; ICDAS-II stage 1: first visible change in enamel; ICDAS-II stage 2: pronounced change in enamel; ICDAS-II stage 3: localized destruction of enamel (without clinically noticeable signs of dentin involvement); ICDAS-II stage 4: underlying dark zone of dentin; ICDAS-II stage 5: large cavity with visible dentin; ICDAS-II stage 6: extensive large cavity with visible dentin. The ICDAS system is also used for the lesion activity assessment (LAA), which is usually based on a combined knowledge of the clinical appearance of the lesion, whether or not there is a lesion in the area of plaque accumulation, and the tactile sensation when a WHO ball-tipped probe is gently passed over the tooth surface. Further information on LAA can be found, for example, in the article by Ekstrand et al. (2007, Oper Dent. 32 (3): 225-235) or the article by Nyvad et al., 2018. Caries Res. 52 (5): 397-405).

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

The detection and evaluation of a carious lesion can be performed by conventional methods for the detection and evaluation of caries. For example, visual inspection, for example with a probe, for example a ball-end probe, or radiographic methods or a combination thereof are widely used. In the context of the present invention, a "cavity-forming carious lesion" may be a carious lesion that can be detected with a ball-end probe, for example a probe having a 0.5 mm diameter sphere at the end (e.g. a WHO probe). Information on the diagnosis and evaluation of dental carious lesions can be found, for example, in the article by Braga et al. (2010, Dent Clin N Am 54: 479-493).

Thus, for example, the present invention provides a composition comprising (a) a self-assembling peptide and (b) a dental substance as defined herein, for use in the treatment of a carious lesion, for example of ICDAS-II stage 1, 2, 3, 4 or 5, preferably a carious lesion progressing to cavity formation, 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. Furthermore, the present invention provides, for example, a kit comprising (a) a self-assembling peptide and (b) a glass ionomer cement-based dental sealant for use in the treatment of a cavity-forming carious lesion, i.e., an ICDAS-II stage 3, 4, 5 or 6 carious lesion, more preferably an ICDAS-II stage 3, 4 or 5 carious lesion, even more preferably an ICDAS-II stage 3 or 4 carious lesion, wherein preferably the carious lesion is active.

Prevention of secondary caries after failure of treatment of a carious lesion proceeding with formation of a cavity using a dental substance means reduction of the occurrence rate of secondary caries. Secondary caries usually occurs under restorations or at the margins of restorations. Failure of treatment using a dental substance can be due to the presence or formation of gaps between the dental substance used to restore the carious lesion proceeding with formation of a cavity (partial failure). It can also be abrasion or loss of the dental substance or restoration partially or completely (complete failure). In one embodiment, the treatment as described herein not only reduces the occurrence rate of secondary caries after failure of treatment using a dental substance, but also increases the time until partial or complete failure of the use of a dental substance, since secondary caries, for example, at the margins of the restoration, can also cause such failure. This is not, however, obligatory, and it is sufficient that secondary caries is not allowed after partial or complete failure of the dental substance.

Restorations classically fabricated with the dental substances defined herein typically fail after a relatively short period of time, which in the art is often associated with secondary caries. For example, fluoride varnish typically lasts up to 3 days under conditions found in the oral cavity, including abrasion during chewing. In protected areas, the varnish may also last longer. Sealants, such as glass ionomer cement-based sealants, typically fail slowly over a period of 2 to 10 years. Bonding agents are typically used to bond a restoration, such as a composite or a crown, to the tooth surface. Failure of the bonding agent and the restoration are typically related to each other. The period of time until failure of a dental substance also depends on several factors in addition to the material, namely the patient (e.g., history of extensive dental treatment due to frequent treatment failures), hygiene, technical expertise of the dentist, location, etc.

In the context of the invention, the applicants have found that the combination of a dental substance with a self-assembling peptide does not allow secondary caries, so that in case of unsuccessful treatment using a dental substance, for example, when a gap is formed between a restoration made of a dental substance or when a dental substance or a restoration associated with a tooth by a dental substance is abraded/fallen out, the matrix formed by the self-assembling peptide, which provides remineralization, protects the tooth from the development of secondary caries. Remineralization of the matrix formed by the self-assembling peptide can already begin when the tooth (i.e., the cavity) is in contact with the self-assembling peptide, and, depending on the availability of ions (e.g., coming from the dental material and/or saliva), under the coating of the dental material. In any case, in case of unsuccessful treatment using a dental material, for example, when a gap is formed between the material and the tooth, the matrix formed by the self-assembling peptides can remineralize and protect the tooth from secondary caries. The self-assembling peptide can also assemble around the margins of restorations made with dental material and protect these areas, which also have a high risk of developing secondary caries.

If said self-assembling peptide and said dental substance are mixed, the dental substance may additionally serve as a reservoir for delayed release of the self-assembling peptide.

Pretreatment with self-assembling peptides has been used to increase bond strength on demineralized dentin (Barbosa-Martins et al. 2018. J Mech Behav Biomed Mater 81:214-221). It is known from the state of the art that bonding of restorations, particularly composites, to dentin is problematic since, compared to enamel, collagen is present as an organic component and the water content in dentin is significantly higher and the ion concentration is significantly lower than in enamel. Barbosa-Martins et al. tested whether pretreatment with self-assembling peptides increased bond strength for two adhesive systems, Adper Single Bond (SB) and Clearfil SE Bond (CSE). Both are polymer-based composites. Benefits were found for bonding to SB but not to CSE. It is believed that this is due to the fact that SB requires etching with phosphoric acid, which is believed to improve the assembly of the self-assembling peptide. In contrast, CSE is a self-etching system that has a significantly lower pH. Due to the significant differences between dentin and enamel surfaces, this publication provides a conclusion only for dentin lesions, but not for enamel lesions. Enamel lesions are preferred for this invention. Said publication also does not indicate whether the results reported are also applicable to dental substances other than composites. In one embodiment of the invention, the dental substance is not a polymer-based composite. In fact, other dental substances, such as glass ionomer cement sealants, have advantageous properties in combination with self-assembling peptides as described herein.

Dental substances as defined herein are well known to those skilled in the art and are commonly available on the market. Dental substances include dental sealants, dental varnishes, such as fluoride varnishes, and dental adhesives. The present invention thus provides a composition or a kit comprising (a) a self-assembling peptide and (b) a dental substance selected from the group consisting of dental restorative materials, such as dental sealants, dental varnishes, and dental adhesives (in the case of a glass ionomer cement-based sealant kit), preferably for use in the treatment of a carious lesion of ICDAS-II stages 1-6, more preferably for use in the treatment of a carious lesion occurring with cavity formation, such as a carious lesion of ICDAS-II stages 3, 4 or 5, as described above, in particular for use in the prevention of secondary caries.

The dental substance used in the present invention may comprise an acidic polymer selected from the group consisting of acrylate and methacrylate, an ionomer, a giomer, an ormocer® and any other suitable polymer and/or a monomeric form thereof. Preferably, said dental substance comprises components that are capable of polymerizing, for example, in a carious lesion occurring with the formation of a cavity or before filling such a lesion. Thus, the components of the dental substance may initially be in monomeric form and may form a restoration (in particular in the case of a sealant) or a protective layer (in particular in the case of a varnish), or be used to bind the restoration (in particular in the case of a binder) upon polymerization of said monomeric components. Said dental substance may be a material suitable for curing with visible light, a self-polymerizing material or a material suitable for curing with UV light, preferably a material suitable for curing with visible light.

In one embodiment of the present invention, the dental substance is a dental varnish, such as a fluoride varnish. Fluoride varnishes are well known to those skilled in the art and are commercially available. Conventional fluoride varnish typically contains fluoride in the form of a salt or a silane preparation in a high concentration in an adhesive solution, such as a fast-drying alcohol and a polymer-based solution. The concentration, type of fluoride and delivery method vary depending on the manufacturer. Common concentrations are 2.5-5% sodium fluoride or about 1% difluorosilane, such as in a polyurethane, polyacrylate, rosin or shellac base. Varnishes may contain CPP-ACP, such as MI Varnish (GC, Japan). Further suitable fluoride varnishes include, for example, Durophat® 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 on fluoride varnishes can be found, for example, in Chu and Lo (Gen Dent. Jul-Aug 2006; 54 (4): 247-253).

In a preferred embodiment, the dental substance 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 in the field of dentistry and are commercially available. Information on dental sealants can be found, for example, in Naaman et al. (Dent J. 2017, 5, 34). In the context of the present invention, dental sealants can be selected from the group consisting of glass ionomer cement-based sealants, polymer-based sealants and modified ceramic materials (e.g. Ormocer ^® ). Glass ionomer cement-based sealants comprise glass ionomer cements, polymer-modified glass ionomer cements. Polymer-based sealants comprise polyacid-modified composite polymers (also called "compomers") and polymer-based composites (also called composite polymers), such as flowable composite polymers.

Preferably, said 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 a kit comprising (a) a self-assembling peptide and (b) a glass ionomer cement-based sealant, such as a glass ionomer cement, a polymer-modified glass ionomer cement or a glass ionomer-type material. Said composition or kit can be used in the treatment of a carious lesion, such as ICDAS-II stages 1-6. Sealants are typically used to treat non-carious lesions that form a cavity and can be used for this purpose in combination with self-assembling peptides, such as P11-4, in mixed form or as a kit, wherein the self-assembling peptide can be applied first, followed by the application of the sealant.

The present invention further opens up the possibility of treating carious lesions occurring with cavity formation, for example stage 3-5 according to ICDAS-II, in particular 3-4, using sealants based on glass ionomer cement in combination with self-assembling peptides, for example P11-4. The advantage of this is that in combination with a self-assembling peptide, secondary caries is prevented (or the incidence of secondary caries is reduced).

Glass ionomer cements belong to a class of materials known as acid-based cements and they are widely used in the field of dentistry and are available in the market. 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 glass ionomer cements in dentistry is "glass polyalkenoate cement". The main ingredients of glass ionomer cement are polymeric water-soluble acid, alkaline (leachable) glass and water. The polymers commonly used in glass ionomer cements are polyalkenoic acids. The glass used for glass ionomer cements is an alkaline glass and thus capable of reacting with acid to form a salt, for example, aluminosilicate glass with fluoride and phosphate additives is often used. The glass available on the market for glass ionomer cements is usually based on calcium and strontium compounds and contains fluoride. Glass ionomer cements usually harden within 2-3 minutes from the start of mixing through an acid-base reaction and are attached to the tooth surface through chemical bonding due to ionic bonds formed between the carboxylate groups on the polyacid molecules and the calcium ions in the tooth surface.

In the context of the present invention, "Glass ^Carbomer® " (GCP Dental, The Netherlands) is also included within the term "glass ionomer cement based sealant". It is a new commercial glass ionomer type material which includes substances which are often not included in glass ionomer compounds, such as silicone oil containing polydimethylsiloxane. Similarly, "Activa" is a vulcanized glass ionomer cement based sealant and is also included within the term "glass ionomer cement based sealant".

Polymer-modified glass ionomer cements can also be used as a dental sealant according to the present invention. These materials contain the same main components as traditional glass ionomers (alkali glass powder, water, polyacid), but also include a monomer component and a related initiator composition. The monomer is typically 2-hydroxyethyl methacrylate (HEMA), and the initiator can be carboquinone or other suitable photoinitiators.

In another embodiment, the dental sealant of the present invention uses polymer-based sealants, i.e., polymer-based dental sealants. Polymer-based sealants are well known in the dental field and are typically based on an oligomeric matrix containing components such as Bisphenol A-glycodyl methacrylate (bisGMA), urethane dimethacrylate (UDMA) or other dimethacrylate monomers (e.g., TEGMA or HDDMA), or on a semicrystalline polyceramic (PEX). They can be divided into four generations, depending on the polymerization method. The first generation was polymerized by ultraviolet rays. The second generation was self-polymerizing polymer-based sealants or chemically cured sealants. In this case, a tertiary amine (activator) is added to one component and mixed with the other component. The reaction between these two components creates free radicals that initiate the polymerization of the polymer sealing material. The third generation comprises visible light polymerizable polymer-based sealants, whereby visible light, typically in the wavelength range of approximately 470 nm, activates photoinitiators (e.g., as explained herein) that are present in the sealant material. The fourth generation of polymer-based sealants are polymer-based fluoride sealants, which are a product resulting from the addition of fluorine-releasing particles to traditional polymer-based sealants. Polymer-based sealants can be filled or unfilled, i.e., the polymer-based sealants can contain a filler, such as described for polymer-based composites herein. Generally, unfilled polymer-based sealants exhibit a lower viscosity. Suitable commercially available polymer-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 present invention provides a composition or kit comprising (a) a self-assembling peptide and (b) a polymer-based sealant, for example, for use in the treatment of a carious lesion. Polymer-based sealants are typically used to treat non-carious lesions that form cavities and can be used for this purpose in combination with self-assembling peptides, such as P11-4.

The present invention further opens up the possibility of treating carious lesions occurring with cavity formation, for example stages 3-5 according to ICDAS-II, in particular 3-4, using polymer-based sealants in combination with self-assembling peptides, such as P11-4. Said advantage is that in combination with a self-assembling peptide, secondary caries is prevented (or the incidence of secondary caries is reduced).

As used herein, "polymer-based sealants" also include polymer-based composites, preferably flowable polymer-based composites (also known as flowable polymer composites). Thus, polymer-based composites, preferably flowable polymer-based composites, can be used in the present invention as dental sealants or prophylactic polymer restorations. In this document, the terms "polymer-based composites" and "composite polymers" are used interchangeably. Polymer-based composites are types of synthetic polymers that are widely used in dentistry as restorative materials, dental adhesives and sealants. Traditional polymer-based composites contain an oligomeric polymer-based matrix and a filler, such as a glass or ceramic filler. Examples of polymer-based oligomer matrices used include bisphenol A-glycidyl methacrylate (bisGMA), urethane dimethacrylate (UDMA) or other dimethacrylate monomers (e.g. TEGMA or HDDMA), or semicrystalline polyceramics (PEX). Glass fillers are typically made of crystalline quartz, silica, lithium barium aluminum glass or zinc strontium lithium borosilicate glass. Ceramic fillers include zirconia silica and zirconia. Composite polymers typically also contain an initiator system, such as a photoinitiator such as camphorquinone, phenylpropanedione (PPD) and lucrin (TPO). These initiator systems initiate the polymerization reaction of the polymers upon application of blue light. They are also chemically cured resin composite systems (also known as "self-curing" resin-based sealants), usually available as a system of 2 pastes (base and catalyst), which begin to harden when the said base and catalyst are mixed. Various additives can be added, for example, to control the reaction rate or the physical characteristics of the dental composite resins, for example, dimethylglycosamine, which can be added to achieve a certain flowability. Dental composite materials demonstrate the ability to adhere to the tooth structure by micromechanical bonding, which is enhanced by acid etching. Very high bond strengths to the tooth structure can be achieved by additional application of dental adhesives.

Polymer-based composites can be classified according to their handling into the categories of "flowable" and "packable". Typically, flowable polymer composites exhibit filler contents below 60% by volume, such as around 35-55% by volume. Packable polymer composites typically exhibit higher filler contents, such as above 60% by volume. Flowable polymer composites generally exhibit lower viscosity and, therefore, favorable wetting properties and easy processability compared to packable polymer-based composites.

Suitable commercially available composite polymers 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 include 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, USA) 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 polymer-based composite, preferably a flowable polymer-based composite, preferably for use in the treatment of a carious lesion, more preferably for use in the treatment of a cavity-forming carious lesion, such as an ICDAS-II stage 3, 4 or 5 carious lesion as described herein.

Polymer-based sealants, as used herein, also include polyacid-modified polymer composite polymers (also referred to as "compomers"). Thus, the dental sealant used in the present invention can be a polyacid-modified composite polymer (also referred to as a "compomer"). Compomers are similar to traditional composite polymers in that their curing reaction is typically an addition polymerization, in that they do not contain water, and in that most of the components are identical. They are typically bulky macromonomers, such as bisglycidyl ether dimethacrylate (bisGMA) or its derivatives, and/or urethane dimethyl acrylate, which is miscible with viscosity-lowering solvents, such as triethylene glycol dimethacrylate (TEGDMA). These polymer systems are filled with non-reactive inorganic powders such as quartz or silicate glass such as _SrAlFSiO4 . These powders are usually coated with silane to promote bonding between the filler and the matrix in the cured material. Compomers typically contain additive monomers that differ from those in traditional composites, which contain acidic functional groups, such as the diester of 2-hydroxymethyl methacrylate in butane tetracarboxylic acid (TGB). The curing reaction is usually light-triggered, but there are also systems where curing occurs by mixing two pastes, each containing a component of the free radical initiator system. Examples of suitable compomers available on the market 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 polymer, preferably for use in the treatment of a carious lesion, more preferably for use in the treatment of a cavity-forming carious lesion, such as an ICDAS-II stage 3, 4 or 5 carious lesion as described herein.

In another embodiment, the dental substance used in the present invention is a dental bonding agent, also called "dentin bonding agents". Dental bonding agents are well known in the dental field and are typically polymeric materials that are traditionally used to provide strong adhesion of dental composite restorative material to both dentin and enamel. Dental bonding agents are often methacrylates with some volatile carrier and a solvent like acetone or ethanol/water. BisGMA or TEGMA are commonly used as adhesives. The application of dental substances typically requires the use of etchants such as phosphoric acid, citric acid/calcium chloride, or oxalic acid/aluminum nitrate, etc., the use of a primer such as NTG-GMMA/BPDM, HEMA/GPDM or 4META. There are available dental bonding systems that combine all the necessary components in a single 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), BeautiBond Generation 7 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 the treatment of a carious lesion, more preferably for use in the treatment of a carious lesion that is progressing with cavity formation, for example, a carious lesion of stage 3, 4 or 5 according to ICDAS-II, as described herein.

In the present application, more than one type of dental substance may be used. For example, the kit of the present application may comprise a dental sealant based on glass ionomer cement and a fluoride varnish. Thus, for example, when using such a kit for treating a carious lesion, the self-assembling peptide may be applied first to said optionally cleaned and/or treated tooth surface, for example, the surface of a carious lesion (either before the fluoride varnish or in the same composition with the fluoride varnish), and the glass ionomer cement may be applied after the application of the fluoride varnish.

In one embodiment of the invention, no fluoride varnish is applied.

In the context of the present invention, it is preferred that said dental substance, for example a glass ionomer cement, a calcium silicate cement, a polyacid-modified composite polymer or a polymer-based sealant, is capable of releasing fluorides. In a particularly preferred embodiment, the dental substance is capable of releasing calcium and fluoride. For example, glass ionomer cements have both properties, since they are based on calcium aluminofluorosilicate glass powder or strontium glass and an acid, typically polyacrylic acid. The dental substance may also, or alternatively, be permeable to ions, for example from saliva, in particular after application to the tooth. Accordingly, substances present in saliva, such as calcium or phosphates, can penetrate into the dental substance. These two characteristics improve the remineralization of the tooth surface, in particular the cavity, even before the dental substance falls out, and thus improve the protection.

There are dental substances, such as dental sealants, that have an alkaline pH, and there are dental substances, such as dental sealants, that have a neutral or acidic pH. The pH of the dental substance can be determined before the material is cured, in particular in an aqueous environment. For example, many cements, such as Portland cement, are alkaline. In the context of the invention, both alkaline and neutral or acidic dental substances can be used, but this can affect the type of self-assembling peptide used, as explained in more detail below.

Portland cement is an example of a calcium silicate cement that can be used in the context of the invention. It is preferably used for dentinal restorations in close proximity to the dental pulp due to the relatively long setting time. For example, a mineral trioxide aggregate (MTA) can be used, such as a fast-hardening mineral trioxide aggregate such as Biodentine (Septodont, Saint Maur des Fosses, France), which sets in about 12 minutes or less; Watson et al., 2014. Dental Mater. 30 (1): 50-61).

The self-assembling peptides used in the present invention are peptides capable of forming three-dimensional "scaffolds", thus promoting tissue regeneration. They can assemble in one dimension to form a beta structure, and into higher-order structures, such as ribbon-like assemblies. Three-dimensional supramolecular structures of self-assembling proteins can be formed that have the ability to bind to calcium phosphate.

In the context of the present invention, the self-assembling peptides may be capable of self-assembling on their own, as is the case, for example, with the peptides P11-4, P11-8, P11-2, P11-5 mentioned below, but they may alternatively be capable of self-assembling in combination of two self-assembling peptides, as is the case, for example, with the peptides P11-13/P11-14 and P11-28/P11-29, P11-30/P11-31 mentioned below.

In the context of the present invention, self-assembling peptides are preferred, as disclosed in patent applications WO 2004/007532 A1, US10/521628, US12/729046, US13/551878, US 14/062768 or WO2014/027012 A1, which are incorporated herein by reference in their entirety. In particular, self-assembling peptides having a net charge of +2 or -2 at pH 7.5 can be used in monomeric or assembled form.

The self-assembling peptides used in the present invention preferably comprise a sequence of the formula X1-X2-X1-X2-X1, wherein X1 is an amino acid having an acidic side chain and X2 is an amino acid having a hydrophobic side chain. For example, the self-assembling peptide used in the present invention comprises a generalized typical sequence of SEQ ID NO: 1, X1-X2-X1-X2-X1, wherein X1 is independently selected from the group consisting of glutamic acid, aspartic acid, 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 at least 80% identical to this sequence. Independently chosen means that, for example, X1 in positions 1, 3 or 5 of the sequence above may differ from each other. Of course, they may also be identical.

Preferably, the self-assembling peptides used in the invention comprise SEQ ID NO: 2, X1-X2-X1-X2-X1, wherein 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.

In a further embodiment, the self-assembling peptides used in the invention may comprise SEQ ID NO: 3, X3-F-X1-W-X1-F-X1, wherein 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, wherein X3 is preferably arginine.

In addition, the self-assembling peptides used in the invention may comprise SEQ ID NO: 4 or, preferably, consist of: X4-X4-X3-F-X1-W-X1-F-X1-X4-X4, wherein X1 is independently selected from the group consisting of glutamic acid and ornithine, X3 is 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. X3 is preferably arginine. Independently, X4 is preferably glutamine.

The self-assembling peptides of the invention may comprise SEQ ID NO: 5, or, preferably, consist of: Q-Q-R-F-X1-W-X1-F-X1-Q-Q, wherein X1 is independently selected from the group consisting of glutamic acid and ornithine.

Preferably, the self-assembling peptides used in the present invention comprise or consist of sequences selected from the general exemplary sequences listed below in Table 1.

Most preferably, said peptides comprise or consist of certain peptides listed in Table 2. Of course, self-assembling peptides assembled in combination with another self-assembling peptide, for example as disclosed herein, can be included in a single kit or in a single composition.

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

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 designated P11-4 and is preferred for the entire invention. In another preferred embodiment, the self-assembling peptide comprises or consists of the sequence of SEQ ID NO: 9 (P11-8).

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

The self-assembling peptides may be modified peptides containing an Ac-N-terminal residue and/or an NH ₂ -C-terminal residue, preferably both, or unmodified peptides. Since unblocked forms tend to initiate a deamination reaction, the terminal residues of all self-assembling peptides of the sequence SEQ ID NO: 1 are preferably blocked to increase stability. In particular, the peptides of the sequence SEQ ID NO: 6, 9, 11, 12, 16 and 17 may contain an Ac-N-terminal residue and an NH ₂ -C-terminal residue. The sequence SEQ ID NO: 18-29 corresponds to modified peptides according to the invention.

Table 1: Summary of typical sequences of preferred self-assembling peptides

SEQ ID NO Peptide name Subsequence Sample SAP SEQ ID NO:1 Generalizing typical sequence 1 X1-X2-X1-X2-X1, where X1 is independently selected from the group consisting of glutamic acid, aspartic acid, glutamine, and ornithine, and X2 is independently selected from the group consisting of alanine, valine, isoleucine, leucine, methionine, phenylalanine, tyrosine, tryptophan, and glutamine P11-2, P11-4, P11-5, P11-8, P11-12, P11-13, P11-14, P11-17, P11-19, P11-20, P11-28, P11-29 SEQ ID NO:2 Generalizing typical sequence 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 P11-4, P11-8, P11-12, P11-13, P11-14, P11-17, P11-28, P11-29 SEQ ID NO:3 Generalizing typical sequence 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, wherein X3 is preferably arginine P11-4, P11-8, P11-12, P11-13, P11-14, P11-17, P11-28, P11-29 SEQ ID NO:4 Generalizing typical sequence 4 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 selected from the group consisting of arginine, glutamic acid and ornithine, X4 is independently selected from the group consisting of glutamine, glutamic acid, serine, threonine and ornithine. X3 is preferably is arginine. Regardless, X4 is preferably glutamine. P11-4, P11-8, P11-12, P11-13, P11-14, P11-17, P11-28, P11-29 SEQ ID NO:5 Generalizing typical sequence 5 Q-Q-R-F-X1-W-X1-F-X1-Q-Q, where X1 is independently selected from the group consisting of glutamic acid and ornithine. P11-4, P11-8

Table 2: Preferred self-assembling peptides. X1 positions are underlined.

SEQ ID NO Peptide name Sequence (Single Letter Code) % amino acid identity P11-4 (ClustalW (2.1, default parameters)) SEQ ID NO:6 P11-4 QQRF E W E F E QQ 100 SEQ ID NO:7 P11-2 QQRF Q W Q F E QQ 81.8 SEQ ID NO:8 P11-5 QQRF O W O F Q QQ 72.7 SEQ ID NO:9 P11-8 QQRF O W O F E QQ 81.8 SEQ ID NO:10 P11-12 SSRF O W O F E SS 45.4 SEQ ID NO: 11 P11-13 EQEF E W E F E QE 72.7 SEQ ID NO:12 P11-14 QQOF O W O F O QQ 63.6 SEQ ID NO:13 P11-17 TTRF E W E F E TT 63.6 SEQ ID NO:14 P11-19 QQRQ O Q O Q E QQ 54.5 SEQ ID NO:15 P11-20 QQRQ E Q E Q E QQ 72.7 SEQ ID NO:16 P11-28 OQOF O W O F O QO 45.4 SEQ ID NO:17 P11-29 QQEF E W E F E QQ 90.9 SEQ ID NO:30 P11-16 NNRF O W O F E NN 45.4 SEQ ID NO:31 P11-18 TTRF O W O F E TT 45.4 SEQ ID NO:32 P11-26 QQOQ O Q O Q O QQ 36.4 SEQ ID NO:33 P11-31 SSOF O W O F O SS 27.3

These self-assembling peptides preferably do not have restriction sites for patient endopeptidases. They also do not need to contain a special cell recognition motif.

In one embodiment, in the kit or composition of the present invention, the self-assembling peptide is predominantly in monomeric form, for example, at least 70%, at least 80% or at least 90% of the self-assembling peptides are present in the monomeric state. For this purpose, if the peptide assembles at a pH of about or below 7.5, the pH of said composition may be higher than said pH, whereby the peptides begin to self-assemble (for example, pH 7.5 for P11-4), preferably 0.1-0.5 pH units higher than said pH, or more than 0.5 pH units higher than said pH. Said pH may be buffered to avoid rapid aggregation. It may be advantageous if aggregation and hydrogel formation begins soon after application to the carious cavity. Accordingly, said pH may be 0.1-0.5 pH units higher than the pH at which the peptide begins to self-assemble without buffering. In one embodiment, said composition may comprise a dried peptide, for example, obtained according to WO 2014/027012.

In another embodiment, a kit or composition of the invention comprises self-assembling peptides in a predominantly assembled form, such as at least 70%, at least 80%, or at least 90% are present in an assembled form, or substantially in an assembled form, and a buffer (at a pH that stabilizes the assembled form). The assembled self-assembling peptide typically forms a hydrogel.

The choice of the assembly state of the protein depends on several factors. A predominantly monomeric form of the self-assembling peptide improves the diffusion of the monomer into the demineralized enamel and may be particularly advantageous for smaller lesions that undergo cavity formation, such as partial cavity lesions or lesions with micro cavities (e.g., ICDAS 1-3). A predominantly assembled form may result in a more rapid action of the peptide and thus improve function, particularly for larger cavities (e.g., ICDAS 4, 5, or 6). If the self-assembling peptide and the dental substance are mixed, the pH required for the dental substance to cure and the ionic strength are usually critical. In the case of a composition comprising both the dental substance and the self-assembling peptide, either a predominantly monomeric self-assembling peptide or a predominantly polymeric self-assembling peptide may be used at the appropriate pH.

Combinations of complementary self-assembling peptides, such as P11-4 and P11-8, provide significantly faster assembly, leading to faster application and stability due to their attraction to each other.

For example, according to the present invention there is provided a composition or a kit comprising (a) a self-assembling peptide comprising a general exemplary sequence according to SEQ ID Nos: 1-5, for example a general exemplary sequence according to SEQ ID NO: 1, preferably a general exemplary sequence according to SEQ ID NO: 2, more preferably a general exemplary sequence according to SEQ ID NO: 3, even more preferably a general exemplary sequence according to SEQ ID NO: 4, and most preferably a general exemplary sequence according to SEQ ID NO: 5, and (b) a dental substance, for example a dental sealant, preferably a glass ionomer cement based sealant, for example for use in the treatment of a carious lesion proceeding with cavity formation, for example a carious lesion of stage 4 or 5 according to ICDAS-I, as described above. In particular, this can be used for the prevention of secondary caries.

In a preferred embodiment, the present invention provides a composition or a kit comprising (a) a self-assembling peptide comprising, preferably consisting of, a sequence selected from the sequences according to SEQ ID NOs: 6-17 and 30-33, or a sequence at least 80%, more preferably at least 90% identical to said sequence, preferably the sequence according to SEQ ID NO: 6 or a sequence at least 80%, more preferably at least 90% identical thereto, and (b) a dental substance, such as a dental sealant, preferably a glass ionomer cement based sealant, preferably for use in the treatment of a carious lesion, both an early and an advanced carious lesion, more preferably for use in the treatment of a carious lesion that is proceeding with cavity formation, such as an ICDAS-II stage 3-5 carious lesion as described above. P-11 is most preferred for use in the context of the present invention. In particular, it can be used to prevent secondary caries.

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

In the context of the invention, the self-assembling peptide used, for example P11-4, can undergo self-assembly at a low pH, in particular at a pH below 7.5. Carious lesions usually have an acidic pH, which ensures the assembly of the self-assembling peptide at the site of the lesion. A preferred peptide that undergoes self-assembly at a low pH, in particular at a pH below 7.5, is P11-4.

The expression that the self-assembling peptides (SAP) undergo self-assembly at a pH below 7.5 means that they have the ability to self-assemble at a pH below 7.5. The ionic strength also influences the state of assembly of the selected self-assembling peptide. Preferably, the self-assembling peptides used in the present invention are capable of self-assembly at a pH below 7.5 and at least a physiological ionic strength. Self-assembling peptides are capable of self-assembly at a pH below 7.5 and at least at a physiological ionic strength may begin to undergo self-assembly at said pH, as is the case, for example, with the preferred peptide, P11-4, but this is not necessary. They may also be capable of being in a self-assembled state at a higher or lower pH.

A person skilled in the art knows how to determine and measure the ionic strength of a solution. The ionic strength I is generally calculated using the formula I=½∑ z _i ² b _i , where z is the valence and bi is the molal concentration [mol/kg{H ₂ O}] of the i-th ionic concentration. The summation of ∑ is taken over all ions in the solution. For example, the ionic strength of a 150 mM NaCl solution is approximately 0.15 mol/l. This is also approximately equal to the ionic strength of blood. The ionic strength of saliva present in the oral cavity is usually significantly lower, for example approximately 0.04 mol/l. In the context of the invention, the ionic strength in the physiological range is taken to correspond to an ionic strength of 0.15 mol/l.

If necessary, the mechanical characteristics can be influenced by the concentration of the self-assembling peptide and additionally by the type of molecules and ions present in the composition. The composition containing the self-assembling peptide used in the invention can, for example, contain NaCl and, optionally, a biologically suitable buffer, such as Tris.

In one embodiment, the assembled state of the self-assembling peptide may also be controlled by ionic strength, for example, assembly may be induced by combination with a buffer having a high ionic strength (e.g., above 0.15 mol/L), which may further comprise calcium, phosphate and/or fluoride ions.

The pH at the site of the lesion (or between the surface of the cavity-forming carious lesion and the dental substance) may depend on the pH of the dental substance. Most dental substances have a neutral or acidic pH, such as chitosan-arginine-amide .

Alternatively, in the context of the invention, the self-assembling peptide used may alternatively self-assemble at a high pH, in particular at a pH of at least 7.5. In this case, the dental substance, for example Portland cement or any calcium hydroxide-based material, typically has a pH of at least 7.5. A preferred peptide that self-assembles at a high pH, in particular at a pH of at least 7.5, is P11-8.

One aspect of the present invention is a composition or kit according to the present invention for use in the treatment of a carious lesion proceeding with the formation of a cavity as described above, preferably a carious lesion proceeding with the formation of a cavity in enamel, in particular for use in preventing or reducing the incidence of secondary caries after failure of the use of a dental substance. Accordingly, the primary carious lesion can be treated, in which the incidence of secondary caries is reduced by the treatment according to the invention. The treatment of a carious lesion in the context of the present invention is typically performed by a dentist.

The treatment may comprise the steps of (a) cleaning the tooth surface, (b) optionally drying the tooth surface, (c) optionally etching the tooth surface, (d) applying the composition or kit components to the carious lesion, and (e) optionally curing, wherein steps (b) and (c), if present, may be performed in any order.

Typically, step (a) precedes steps (b-e) of the treatment procedure. Steps (b-d) are more flexible in terms of their sequence during application, and step (e) typically follows steps (b-d). Thus, for example, etching (if applied) can be performed before or after drying, or in the middle of step (d), etc. The sequence and application of said steps typically depends on the type of dental substance used in the present invention and corresponds to the sequence and application of the steps typically applied to said dental substance, if said dental substance is used without a self-assembling peptide. For example, if a polymer-based sealant is used as the dental substance, it is preferable that the tooth surface is etched before applying the polymer-based sealant. Polymer-based sealants are also more sensitive to moisture than glass ionomer cements. Thus, in the case of applying polymer-based sealants, drying the tooth surface is preferable. Preferably, in this case, the tooth is both dried and etched. The dentist is aware of the recommended treatment steps for the various dental substances.

Step (a), "cleaning the tooth surface", means that at least the surface of the carious lesion is cleaned, i.e. substances, such as food particles, dental plaque, dental plaque or dental films, etc., are removed from the tooth surface, at least from the surface of the carious lesion, by suitable methods. Such cleaning methods may include treatment with a traditional toothbrush, treatment with special toothbrushes, such as plaque brushes, treatment with dental scalers and curettage spoons, the use of mouth rinses, such as the use of antibacterial mouth rinses, etc. The cleaning preferably includes the removal of plaque. The cleaning may further include a partial or complete removal of tissue destroyed by caries, such as by cleaning, drilling, sandblasting or laser burning, by using a dental excavator, or by chemomechanical removal (for example using the Carisolv ^® system). Removal of dental tissue destroyed by caries is generally not required in the context of the invention, and drilling is preferably not performed.

In this case, in the case of carious lesions in more advanced stages of caries, for example, carious lesions of stages 3, 4, 5 and 6 according to ICDAS-II, it may be preferable to at least partially remove the destroyed dental tissue. In the case of performing the removal of destroyed dental tissue, it is preferable that the carious dental tissue is removed by a "gentle" method, for example by using a dental excavator or a chemomechanical removal method, but not by drilling. The inventors have found that, in the context of enamel lesions without dentine involvement (as is preferred herein), it is not necessary to completely remove the carious tissue. It is particularly preferable that intact tissue is not removed during the removal of carious tissue, which is usual in the case where drilling is used as a removal technique. In the case of lesions with dentine involvement, the carious tissue is preferably removed.

As described above, the application of step (b), i.e. drying of the dental surface, may be preferred for the application of specific dental substances, such as polymer-based sealants, dental adhesives or polymer-based composites. Drying may be performed before etching (if etching is performed), after etching, or in the context of step (d), such as between the application of the self-assembling peptide and the application of the dental substance. Drying of the dental surfaces is usually performed using an air stream.

Furthermore, step (c), i.e. etching the tooth surface, may be advantageous for the application of specific dental substances, such as polymer-based sealants, dental adhesives or polymer-based composites. Etching is a standard method traditionally used in dental filling procedures. "Etching" means creating a microscopic porosity on the tooth surface into which the self-assembling peptide and/or the dental substance, such as a dental sealant, can flow, thereby increasing retention, increasing the surface area and increasing the bonding strength between the dental substance and the tooth surface. Etching substances that can 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 substance is phosphoric acid, which is often also called a conditioning agent. Typically, the etching agent is applied to the cleaned and optionally dried tooth surface for a period of 10-60 seconds, such as 15-30 seconds. After etching, the tooth surface is preferably rinsed, such as with water. Depending on the dental substance applied, the tooth surface is dried after an optional rinsing step before applying the dental substance, such as in the case of polymer-based sealants. In one embodiment, etching is performed after applying the self-assembling peptide and before applying the dental substance.

Step (d) comprises applying the composition or components of the kit to the carious lesion, preferably a carious lesion that is undergoing cavity formation. This means that the composition or components of the kit are applied to the carious lesion by suitable methods known to the dentist, for example by filling. When the kit of the invention is used to treat a carious lesion, preferably step (d) comprises (i) applying the self-assembling peptide or a composition comprising the self-assembling peptide (contained in the kit) to the carious lesion, and then (ii) applying a dental substance to the carious lesion. "Applying a dental substance" generally refers to application when it is professionally and conventionally performed by a dentist, for example according to the manufacturer's instructions. The self-assembling peptide can be applied, for example, as a solution in a suitable buffer. The self-assembling peptide can be applied to the carious lesion by means of any suitable applicator, for example a syringe, a sponge, etc. For example, application can be carried out using Curodont® Repair. The kit preferably also contains an applicator.

In one embodiment, step (d) comprises applying a mixture of the self-assembling peptide and the dental substance to the carious lesion undergoing cavity formation. This mixture may be a composition of the present invention or components of a kit of the present invention mixed before or during application. The mixture may be applied in a manner conventional in the dental art for the corresponding dental substance, for example, according to the manufacturer's instructions.

The use of optional step (e), i.e. curing, depends on the dental substance being used. Most dental substances require some form of curing. The term "curing" refers to a process in which a chemical reaction (e.g. polymerization) or physical action (e.g. evaporation) occurs resulting in a harder, stronger, or more stable bond (e.g. bonding) or substance (e.g. cured polymer). Typical curing for a dental material is chemical curing, curing by the application of light, thermal curing, or a combination of these.

Thus, the present invention provides, for example, a composition or a kit according to the present invention for use in the treatment of a carious lesion, preferably a carious lesion progressing to cavity formation, for example a carious lesion of stage 3, 4 or 5 according to ICDAS-II as described above, wherein the treatment of the carious lesion comprises the steps of (i) cleaning the dental surface by removing plaque, in particular cleaning the surface of the carious lesion, optionally including complete or partial removal of the affected tissue, wherein preferably no drilling is performed, (ii) applying a composition comprising a self-assembling peptide as described above to the carious lesion, for example a self-assembling peptide comprising the general exemplary sequence according to SEQ ID Nos: 1-5 (for example P11-4), (iii) applying a dental substance to the carious lesion, for example a glass ionomer cement sealant or fluoride varnish, and (iv) curing fluoride varnish, for example by drying, or a glass ionomer cement sealant, for example by chemical curing or a combination of chemical curing and visible light curing. The sequence of steps may be (i) to (iv).

Another example for a composition or kit for use according to the present invention is a composition or kit according to the present invention for use in the treatment of a carious lesion, preferably a carious lesion progressing to form a cavity, for example a carious lesion of stage 3, 4 or 5 according to ICDAS-II as described above, wherein the treatment of the carious lesion comprises the steps of (i) cleaning the tooth surface by removing plaque, in particular, optionally, cleaning the surface of the carious lesion, which includes removing the diseased tissue partially or completely, wherein preferably no drilling is performed, (ii) optionally drying the surface of the carious lesion, (iii) applying an etching agent, for example phosphoric acid, to the surface of the carious lesion for a suitable time and thereby etching the surface of the tooth having the carious lesion, (iv) removing the etching agent, for example by rinsing, (v) applying a composition comprising a self-assembling peptide as described above to a carious lesion, for example a self-assembling peptide comprising a generic exemplary sequence according to SEQ ID NOs: 1-5 (for example P11-4), (vi) drying the surface of a tooth having a carious lesion, (vii) applying a polymer-based sealant, preferably according to the manufacturer's instructions, and (viii) curing the polymer-based sealant with a suitable curing method.

The choice of dental substances in the context of the invention may depend on the type of carious lesion to be treated. For example, the invention advantageously provides the possibility of using a dental sealant for carious lesions occurring with the formation of a cavity of any size or for occlusal carious lesions in pits and/or fissures of a tooth.

Additionally, the present invention provides a method for treating a carious lesion that forms a cavity, comprising the step of applying said components of a kit or a composition of the invention to said carious lesion. For example, said method may comprise steps (a) to (e) as described above, i.e., (a) cleaning a tooth surface, (b) optionally drying the tooth surface, (c) optionally etching or conditioning the tooth surface, (d) applying the composition or components of the kit to the carious lesion, and (e) optionally curing, as described above for the composition and kit for use in treating a carious lesion.

The present invention also solves an additional technical problem. In particular, during the removal of carious dentin from a carious lesion, the dental pulp may be exposed or may be near the lesion, the risk of pulpitis or pulp retraction increases due to an adverse reaction of the pulp to the dental material (sealant, bonding agent, filling agent, polymer, etc.). The inventors have surprisingly found that in order to prevent damage to the pulp when placing a dental restoration near the pulp, the dentist can use a kit or composition according to the invention. Thus, the invention provides a composition or kit according to the present invention for use in pulp capping. The dental material can be dental substances as described above, for example, a dental sealant or bonding agent, or a filling agent, etc. It may be a glass ionomer cement based sealant selected from the group comprising glass ionomer cements and polymer-modified glass ionomer cements, or a polymer-based sealant selected from the group comprising polyacid-modified composite polymers and polymer-based composites. In this context, polymer-based composites are preferred dental materials.

For example, if, in the context of cleaning, in particular removing dentin from a carious lesion, the pulp is exposed or is about to be exposed (to an extent that one skilled in the art may determine increases the risk of pulp retraction or pulpitis), a self-assembling peptide (e.g., P11-4 or a mixture of P11-4 and P11-8, preferably P11-4) in solution can be applied to protect the pulp and enhance remineralization, and then a dental material, such as a dental sealant, is applied.

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 can be mixed prior to application, which is most suitable if the dental material is also biocompatible with the pulp (e.g. MTA).

The composition or kit of the present invention can be used for pulp capping.

The pulp capping may be a direct or indirect pulp capping. The remineralization of a thin layer of softened dentine is particularly advantageous for indirect pulp capping, which can be improved by the self-assembling peptide used in the context of the invention. Glass ionomer or polymer-modified glass ionomers can also be used in the context of the pulp capping according to the invention, in particular for indirect pulp capping. If necessary, different layers of dental sealant can be applied over the pulp capping.

If the self-assembling peptide is placed on top of the pulp to protect the pulp from retraction due to toxicity of the dental filling material, one source of calcium ions for remineralization may be the pulp. In the case of pulp capping, the ions for remineralization come from the pulp (or are delivered to the pulp by the blood), and remineralization can begin immediately after placement (without/before filling failure or formation of micro-gaps or micro-leaks). Of course, additional ions, including fluoride, may come from the dental material. Thus, fluoride-releasing dental materials, such as glass ionomer cements, are also preferred in this context.

The present invention is illustrated, but not limited, by the following examples. All references cited herein are incorporated herein in their entirety.

Description of drawings

Fig. 1.1. Treatment of carious lesion occurring with partial cavity formation (class 3 according to ICDAS).

A: Cross-section of a tooth with partial cavity formation on the left.

B: After cleaning, in this case without removing the softened dentin, applying a solution of mainly monomeric self-assembling peptide (e.g. P11-4, strips) to the tooth surface. The self-assembling peptide begins to diffuse into the cavity.

C: A self-assembling peptide, such as P11-4, assembles into the cavity due to the low pH created by the matrix.

D: The tooth surface is dried out, leaving ribbons & fibers of collected P11-4 inside the lesion.

E: After the solution is absorbed into the cavity, a sealant such as glass ionomer cement (GIC) is applied to allow diffusion of ions (calcium and phosphate) from saliva and from the glass ionomer cement into the lesion.

F: Mineralization (hexagons) in the damage protect the damaged sealant, resulting in a mineralized structure under the damage (protected by the sealant, e.g. GIC).

After damage to the GIC, a completely regenerated tooth surface and structure remains.

Fig. 1.2:

A: Cross-section of a tooth with a partially formed cavity on the left.

B: Carious lesion after removal of softened dentin.

C: Application of monomeric P11-4 (strips) either simultaneously with the filling material or sequentially.

D: Tooth after removal of softened dentin with filling material (gray) and fibrillar P11-4 (longer stripes) at the interface between the tooth and filling material.

E: Mineralization (hexagons) in the micro-cracks between the filling material and the tooth. Calcium phosphate comes either from the filling material (in the case of glass ionomer cement), saliva or from the pulp (via dental lymph).

Fig. 2. Treatment of a cavity-forming carious lesion (ICDAS class 4) using the composition of the invention in a caries model as described in Ex. 2. The right and left columns show two representative examples.

A, B: Drilling of an artificial cavity.

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).

Fig. 3. Treatment of a cavity-forming carious lesion (ICDAS class 4) using the kit according to the invention in a caries model as described in Ex. 3. The right and left columns show two typical examples.

A, B: Drilling of an artificial cavity.

C, D: Application of glass ionomer cement (Aqual Ionofill Plus, VOCO) mixed with 1 drop of P11-4, 10 mg/ml, pH 8.

Fig. 4. Release curve of model substance Congo Red and P11-4 from varnish, glass ionomer cement and Paro-Amin Fluor Gelée. The experiment was performed as described in Examples 6 and 7. Release was measured as described and expressed in arbitrary units (AU). Fig. 4A shows the release of Congo Red from Duraphat varnish (sample A1) and Aqua Ionofil Plus glass ionomer cement (sample B1), and Fig. 4B shows the release of Congo Red from paro Amin Fluor Gelée (sample 2). Fig. 4C shows the release of monomeric P11-4 from Duraphat varnish (sample AP) and Aqua Ionofil Plus glass ionomer cement (sample BP).

Examples

Example 1

A lesion with partial cavity formation (ICDAS class 3) is cleaned. This occurs either with the removal of carious dentin, (Fig. 1.2) or without it (Fig. 1.1). After cleaning, which does not necessarily include the removal of softened dentin, a self-assembling peptide (e.g. P11-4) is applied to the cleaned tooth surface, in particular into the cavity. The self-assembling peptide begins to diffuse into the cavity. In this case, the self-assembling peptide assembles due to the low pH, creating a matrix. After the solution is absorbed into the dentin or enamel (10 sec-5 min, depending, for example, on the surface (in dentin there is a faster absorption than in enamel), a sealant is applied, for example a glass ionomer cement, which ensures the diffusion of ions (Ca and phosphate) from saliva or glass ionomer into the damaged area. This ensures the beginning of mineralization in the matrix protected by the sealant. After the tooth loses the sealant, usually after several months or years, a completely regenerated tooth surface and structure remains, in the absence of secondary caries.

Example 2

An artificial ICDAS class 4 lesion created by drilling into an extracted bovine incisor was treated with the kit of the invention, specifically by applying 1 drop of P11-4, 10 mg/ml, pH 8, followed by glass ionomer cement (Aqual Ionofill Plus, VOCO) according to the manufacturer's instructions. Briefly, a polyacrylic acid conditioner was applied for 10 seconds, rinsed, lightly dried, glass ionomer applied and the material cured before the final procedures. The main steps are shown in Fig. 2. The glass ionomer cement cured to give a hard restoration indistinguishable from one placed without the combination of P11-4 (data not shown). No differences could be detected within a week after application.

Example 3

An artificial ICDAS class 4 lesion created by drilling into an extracted bovine incisor was treated with the composition of the invention. Glass ionomer cement powder (Aqual Ionofill Plus, VOCO) was prepared according to the manufacturer's instructions. 1 mg P11-4 per 100 mg (10 mg per 1 ml powder) was added and mixed with water. The powder was diluted according to the manufacturer's recommendations. The solution was mixed for 30 s and applied to the tooth surface as recommended by the manufacturer (as described above). The main steps are shown in Fig. 3.

The glass ionomer cement was cured to produce a hard restoration indistinguishable from one placed without the combination of P11-4 (data not shown). No differences could be detected within one week of placement.

Example 4 - Prevention of secondary caries

Teeth containing ICDAS class 4 artificial lesions created by drilling into an extracted bovine incisor were treated with a dental sealant (e.g. glass ionomer (e.g. Aqual Ionofill Plus, VOCO)) in the absence or presence of P11-4 according to three different protocols:

1) Dental sealant is applied to the cleaned cavity surface according to the manufacturer's instructions.

2) A drop of P11-4, 10 mg/ml, pH 8, was applied to the cleaned cavity surface so as to cover the said cavity surface. After the solution was substantially absorbed by the tooth surface, the dental sealant was applied to the cleaned cavity surface according to the manufacturer's instructions.

3) Glass ionomer cement powder (Aqual Ionofill Plus, VOCO) was prepared according to the manufacturer's instructions. 1 mg P11-4 per 100 mg was added (10 mg per 1 ml powder) and mixed with water. The powder was dissolved according to the manufacturer's recommendations. The solution was mixed for 30 s and applied to the tooth surface according to the manufacturer's recommendations (as described above).

The teeth were stored in distilled water for 24 hours. The teeth were then subjected to cyclic heat treatment (500 cycles, 5-55°C), immersed in alkaline fuccin, sectioned, and analyzed for dye penetration and remineralization using a stereomicroscope (e.g., as described by Hepdeniz et al., 2016. Eur J Dent. 10(2):163-169).

The formation of microcracks or microleaks and the shrinkage of the dental sealant were monitored over time and remineralization at the site of microcracks was analyzed. Different groups were compared.

Example 5 - Pulp capping - remineralization by pulp and calcium phosphate from the blood

The cavity, which had exposed pulp due to dental caries, was cleaned using an excavator. The pulp was coated with P11-4 solution (e.g. 100 µl, 10 mg/ml pH 8) before placement of dental material, e.g. resin composite, according to the manufacturer's instructions (e.g. layering and light curing, depending on requirements).

There were no pulpal symptoms (i.e. no pulpitis), and pulp retraction was assessed during a follow-up radiographic examination after 1 year.

Example 6 - Release Curve

As is well known to those skilled in the art, the release curve of a substance, i.e. fluoride or 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 experiment, the release curve of a model substance simulating a self-assembling peptide, Congo Red, was evaluated after incorporation into a dental material.

The material was prepared and applied according to the instructions for use (IFU), which usually describes the use of 0.5-0.75 ml of the respective products. Phosphate buffered saline (PBS) was used to simulate saliva. PBS corresponds to the osmolarity and ion concentration of the human body - it is isotonic. To simulate a constant flow of saliva, the taken amount was placed in a solution equivalent to the flow of saliva in 1 hour, which was 20 ml. The diffusion of the material was assessed at 500 nm using a photometer.

Material

Name Supplier Product ID Party Sample ID Duraphat
(Duraphat® dental suspension, varnish) Colgate Na 227955 A Aqua Ionofil Plus
(glass ionomer restorative material) Voco 1513 1827129 B Paro-Amin Fluor Gel Esro -- 1919-1 2 PBS-Taps Sigma P4417 Water Credentials -- -- Oligopeptide-104 Bachem E104 1000020803 -- Congo Red Sigma C6767 BCBR1055 Ethanol Credentials -- -- Syringe 1 ml Codan -- -- Balance Mettler Toledo AE200 -- Ultraviolet VI Pharmacia LKB Ultraspec II -- Ultraviolet cuvette Brand -- -- Spatula Object glasses VWR -- -- Eppendorf tube 2 ml VWR -- -- 50 ml centrifuge tube VWR -- --

Release curve

Preparation of varnish (Duraphat) and paro Amin Fluor Gelée

According to IFU suppliers, it is recommended to add 0.5-1 ml of varnish to the caries site. The concentration of Congo Red was chosen to obtain a clear signal in ultraviolet Vi.

Place 12 mg of Congo Red in a 2 ml Eppendorf tube.

Add 2 ml of varnish directly into the test tube using a syringe.

Vigorously mix Congo Red and varnish until the contents are uniform.

The product was drawn up using a syringe.

Place 0.5-0.75 ml of the product on a glass slide as a thin film - as recommended by IFU.

Allowed to air dry - according to IFU.

Preparation of Ionofil (glass ionomer restorative material).

Placed 12 mg of Congo Red directly into the dental cup.

Added 2 spoons of Ionofil Plus according to IFU.

Add 2 drops of water and mix according to IFU.

Place a quantity of the product on a glass slide.

Endurance.

A PBS solution with 1% EtOH was prepared for preservation.

Place the glass in a 50 ml centrifuge tube.

Added 20 ml PBS.

Placed on the carousel table.

Measurement and calculation.

1 ml of the aged liquid was directly filtered into the UV-Cell.

Measured at 500 nm, control/sample=PBS with 1% EthOH.

The control without Congo Red was subtracted from the measured value.

Results and discussion

Regardless of the material composition, the model substance was easily incorporated into the products. This is demonstrated by visual assessment of the homogeneity of the content, as the homogeneous red/brown distribution is noticeable.

The model substance was also able to diffuse from all three fluorine-containing preparations into artificial saliva. Depending on the composition, the release curve of individual formulations differed.

The model substance Congo red was released from Duraphat varnish and glass ionomer cement (Fig. 4A) and from paro Amin Fluor Gelée used for comparison (Fig. 4B). The release from the varnish was faster than from the glass ionomer cement. It can be seen that the release from the glass ionomer cement was constant and increased over time.

Example 7 - P11-4 Release Curve

Example 7 was performed essentially as Example 6, but with the self-assembling peptide P11-4, unless otherwise described. 12 mg P11-4 (credentis AG) in monomeric form was added to either Duraphat varnish (Sample ID: AP) or Aqua Ionofil Plus glass ionomer cement (Sample ID: BP) in the amounts and under the conditions described above. The curing was performed in a static manner at room temperature. The measurement was performed at 280 nm and the control value for the buffer without peptide was subtracted from the measured value.

The results are shown in Fig. 4C. In support of the previously obtained data, the experiment showed an unexpectedly good diffusion of the peptide from the tested matrix, i.e. Duraphat (sample AP) or Aqua Ionofil Plus (BP), confirming the data obtained with the model substance.

P11-4 was released slowly and essentially continuously from both the varnish and the glass ionomer cement. The amount of peptide released increased over time, as measured over several days. Release from the varnish was faster than release from the glass ionomer cement. Thus, depending on the formulation, slow release was obtained by using a sealant, such as glass ionomer cement, or “fast” release was obtained by using a varnish, such as a rosin-containing formulation.

Rapid release, as seen with a varnish such as Duraphat, may have a beneficial effect on the effectiveness of the peptide shortly after application, which may be particularly useful, for example, for remineralization of varnished lesions.

On the other hand, a longer release may additionally be particularly advantageous for the prevention or treatment of secondary lesions - which may develop slowly over time - under or at the margins of the sealant on healed (filled) lesions. It can be reasonably concluded that the substance will be released from a sealant, such as glass ionomer cement, over a longer period than from a varnish. This is appropriate since the average residence time of a sealant, such as glass ionomer cement, on the tooth is also longer than for a varnish. In addition, the self-assembling peptide released from the filling can remain in place for a long time.

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LIST OF SEQUENCES

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<223> ornithine

<400> 14

Gln Gln Arg Gln Xaa Gln Xaa Gln Glu Gln Gln

1 5 10

<210> 15

<211> 11

<212> Protein

<213> Artificial sequence

<220>

<223> P11-20

<400> 15

Gln Gln Arg Gln Glu Gln Glu Gln Glu Gln Gln

1 5 10

<210> 16

<211> 11

<212> Protein

<213> Artificial sequence

<220>

<221>MOD_RES

<222> 1..1

<223> ornithine

<220>

<223> P11-28

<220>

<221>MOD_RES

<222> 3..3

<223> ornithine

<220>

<221>MOD_RES

<222> 5..5

<223> ornithine

<220>

<221>MOD_RES

<222> 7..7

<223> ornithine

<220>

<221>MOD_RES

<222> 9..9

<223> ornithine

<220>

<221>MOD_RES

<222> 11..11

<223> ornithine

<400> 16

Xaa Gln Xaa Phe Xaa Trp Xaa Phe Xaa Gln Xaa

1 5 10

<210> 17

<211> 11

<212> Protein

<213> Artificial sequence

<220>

<223> P11-29

<400> 17

Gln Gln Glu Phe Glu Trp Glu Phe Glu Gln Gln

1 5 10

<210> 18

<211> 11

<212> Protein

<213> Artificial sequence

<220>

<221>MOD_RES

<222> 1..1

<223> ACETYLATION

<220>

<223> P11-4mod

<220>

<221>MOD_RES

<222> 11..11

<223> AMIDATION

<400> 18

Gln Gln Arg Phe Glu Trp Glu Phe Glu Gln Gln

1 5 10

<210> 19

<211> 11

<212> Protein

<213> Artificial sequence

<220>

<221>MOD_RES

<222> 1..1

<223> ACETYLATION

<220>

<223> P11-2mod

<220>

<221>MOD_RES

<222> 11..11

<223> AMIDATION

<400> 19

Gln Gln Arg Phe Gln Trp Gln Phe Glu Gln Gln

1 5 10

<210> 20

<211> 11

<212> Protein

<213> Artificial sequence

<220>

<221>MOD_RES

<222> 1..1

<223> ACETYLATION

<220>

<223> P11-5mod

<220>

<221>MOD_RES

<222> 5..5

<223> ornithine

<220>

<221>MOD_RES

<222> 7..7

<223> ornithine

<220>

<221>MOD_RES

<222> 11..11

<223> AMIDATION

<400> 20

Gln Gln Arg Phe Xaa Trp Xaa Phe Gln Gln Gln

1 5 10

<210> 21

<211> 11

<212> Protein

<213> Artificial sequence

<220>

<221>MOD_RES

<222> 1..1

<223> ACETYLATION

<220>

<223> P11-8mod

<220>

<221>MOD_RES

<222> 5..5

<223> ornithine

<220>

<221>MOD_RES

<222> 7..7

<223> ornithine

<220>

<221>MOD_RES

<222> 11..11

<223> AMIDATION

<400> 21

Gln Gln Arg Phe Xaa Trp Xaa Phe Glu Gln Gln

1 5 10

<210> 22

<211> 11

<212> Protein

<213> Artificial sequence

<220>

<221>MOD_RES

<222> 1..1

<223> ACETYLATION

<220>

<223> P11-12mod

<220>

<221>MOD_RES

<222> 5..5

<223> ornithine

<220>

<221>MOD_RES

<222> 7..7

<223> ornithine

<220>

<221>MOD_RES

<222> 11..11

<223> AMIDATION

<400> 22

Ser Ser Arg Phe Xaa Trp Xaa Phe Glu Ser Ser

1 5 10

<210> 23

<211> 11

<212> Protein

<213> Artificial sequence

<220>

<221>MOD_RES

<222> 1..1

<223> ACETYLATION

<220>

<223> P11-13mod

<220>

<221>MOD_RES

<222> 11..11

<223> AMIDATION

<400> 23

Glu Gln Glu Phe Glu Trp Glu Phe Glu Gln Glu

1 5 10

<210> 24

<211> 11

<212> Protein

<213> Artificial sequence

<220>

<221>MOD_RES

<222> 1..1

<223> ACETYLATION

<220>

<223> P11-14mod

<220>

<221>MOD_RES

<222> 3..3

<223> ornithine

<220>

<221>MOD_RES

<222> 5..5

<223> ornithine

<220>

<221>MOD_RES

<222> 7..7

<223> ornithine

<220>

<221>MOD_RES

<222> 9..9

<223> ornithine

<220>

<221>MOD_RES

<222> 11..11

<223> AMIDATION

<400> 24

Gln Gln Xaa Phe Xaa Trp Xaa Phe Xaa Gln Gln

1 5 10

<210> 25

<211> 11

<212> Protein

<213> Artificial sequence

<220>

<221>MOD_RES

<222> 1..1

<223> ACETYLATION

<220>

<223> P11-17mod

<220>

<221>MOD_RES

<222> 11..11

<223> AMIDATION

<400> 25

Thr Thr Arg Phe Glu Trp Glu Phe Glu Thr Thr

1 5 10

<210> 26

<211> 11

<212> Protein

<213> Artificial sequence

<220>

<221>MOD_RES

<222> 1..1

<223> ACETYLATION

<220>

<223> P11-19mod

<220>

<221>MOD_RES

<222> 5..5

<223> ornithine

<220>

<221>MOD_RES

<222> 7..7

<223> ornithine

<220>

<221>MOD_RES

<222> 11..11

<223> AMIDATION

<400> 26

Gln Gln Arg Gln Xaa Gln Xaa Gln Glu Gln Gln

1 5 10

<210> 27

<211> 11

<212> Protein

<213> Artificial sequence

<220>

<221>MOD_RES

<222> 1..1

<223> ACETYLATION

<220>

<223> P11-20mod

<220>

<221>MOD_RES

<222> 11..11

<223> AMIDATION

<400> 27

Gln Gln Arg Gln Glu Gln Glu Gln Glu Gln Gln

1 5 10

<210> 28

<211> 11

<212> Protein

<213> Artificial sequence

<220>

<221>MOD_RES

<222> 1..1

<223> ACETYLATION

<220>

<221>MOD_RES

<222> 1..1

<223> ornithine

<220>

<223> P11-28mod

<220>

<221>MOD_RES

<222> 3..3

<223> ornithine

<220>

<221>MOD_RES

<222> 5..5

<223> ornithine

<220>

<221>MOD_RES

<222> 7..7

<223> ornithine

<220>

<221>MOD_RES

<222> 9..9

<223> ornithine

<220>

<221>MOD_RES

<222> 11..11

<223> ornithine

<220>

<221>MOD_RES

<222> 11..11

<223> AMIDATION

<400> 28

Xaa Gln Xaa Phe Xaa Trp Xaa Phe Xaa Gln Xaa

1 5 10

<210> 29

<211> 11

<212> Protein

<213> Artificial sequence

<220>

<221>MOD_RES

<222> 1..1

<223> ACETYLATION

<220>

<223> P11-29mod

<220>

<221>MOD_RES

<222> 11..11

<223> AMIDATION

<400> 29

Gln Gln Glu Phe Glu Trp Glu Phe Glu Gln Gln

1 5 10

<210> 30

<211> 11

<212> Protein

<213> Artificial sequence

<220>

<223> P11-16

<220>

<221>MOD_RES

<222> 5..5

<223> ornithine

<220>

<221>MOD_RES

<222> 7

<223> ornithine

<400> 30

Asn Asn Arg Phe Xaa Trp Xaa Phe Glu Asn Asn

1 5 10

<210> 31

<211> 11

<212> Protein

<213> Artificial sequence

<220>

<223> P11-18

<220>

<221>MOD_RES

<222> 5..5

<223> ornithine

<220>

<221>MOD_RES

<222> 7

<223> ornithine

<400> 31

Thr Thr Arg Phe Xaa Trp Xaa Phe Glu Thr Thr

1 5 10

<210> 32

<211> 11

<212> Protein

<213> Artificial sequence

<220>

<223> P11-26

<220>

<221>MOD_RES

<222> 3

<223> ornithine

<220>

<221>MOD_RES

<222> 5

<223> ornithine

<220>

<221>MOD_RES

<222> 7

<223> ornithine

<220>

<221>MOD_RES

<222> 9

<223> ornithine

<400> 32

Gln Gln Xaa Gln Xaa Gln Xaa Gln Xaa Gln Gln

1 5 10

<210> 33

<211> 11

<212> Protein

<213> Artificial sequence

<220>

<223> P11-31

<220>

<221>MOD_RES

<222> 3

<223> ornithine

<220>

<221>MOD_RES

<222> 5

<223> ornithine

<220>

<221>MOD_RES

<222> 7

<223> ornithine

<220>

<221>MOD_RES

<222> 9

<223> ornithine

<400> 33

Ser Ser Xaa Phe Xaa Trp Xaa Phe Xaa Ser Ser

1 5 10

<---

Claims (24)

1. A composition for the prevention of secondary caries after unsuccessful use of a dental substance in the treatment of a carious lesion occurring with the formation of a cavity, containing: a) a self-assembling peptide, wherein said self-assembling peptide is P11-4 having SEQ ID NO: 6, and b) a dental substance selected from the group consisting of a dental sealant selected from the group of glass ionomer cements and polymer-modified glass ionomer cements containing an acrylate polymerizing component, and a fluoride varnish, wherein said fluoride varnish contains ethyl alcohol as a solvent in combination with rosin or shellac base. 2. The composition according to claim 1, wherein said composition is a pharmaceutical composition. 3. The composition according to claim 1, wherein the dental substance is a fluoride varnish. 4. A kit for the prevention of secondary caries after unsuccessful use of a dental substance in the treatment of a carious lesion that occurs with the formation of a cavity, containing in separate form: a) a self-assembling peptide, wherein said self-assembling peptide is P11-4 having SEQ ID NO: 6, and b) a dental substance which is a dental sealant based on a glass ionomer cement selected from the group consisting of glass ionomer cements and polymer-modified glass ionomer cements containing an acrylate polymerizing component. 5. The kit according to item 4, wherein said kit is a pharmaceutical kit. 6. The kit according to item 4, wherein the dental substance contains a non-polar or polar organic solvent. 7. The composition according to any one of claims 1-3 or the kit according to any one of claims 4-6, wherein said dental substance has the ability to release fluoride ions, wherein preferably said dental substance also has the ability to release calcium ions. 8. The composition according to any one of claims 1-3 or 7 or the kit according to any one of claims 4-7, wherein the dental substance is permeable to and from ions. 9. The composition according to any one of claims 1-3 or 7-8 or the kit according to any one of claims 4-8, wherein the self-assembling peptide is mainly in the form of a monomer. 10. Use of a composition according to any of claims 1-3 or 7-9 or a kit according to any of claims 4-9 in the prevention of secondary caries after unsuccessful use of a dental substance in the treatment of a carious lesion occurring with the formation of a cavity. 11. The use according to item 10, wherein the treatment of the carious lesion includes the steps of: a) cleaning the tooth surface, b) optionally, drying the tooth surface, c) optionally, etching of the tooth surface, d) applying the said composition or the said components of the kit to the carious lesion, e) optionally, curing. 12. Use of a composition according to any of claims 1-3 or 7-9 or a kit according to any of claims 4-9 in a protective pulp coating. 13. Use according to claim 12, wherein said composition or said components of the kit are applied to the carious lesion upon contact with the pulp. 14. Use according to any one of claims 10 or 12, 13, wherein applying said components of the kit to the carious lesion comprises applying the self-assembling peptide to the carious lesion and then applying said dental substance to the carious lesion. 15. Use according to any one of claims 10 or 12, 13, wherein applying said components of the kit to the carious lesion comprises applying a mixture of said self-assembling peptide and said dental substance to the carious lesion.