WO2025059523A2 - Peptide-polymer adhesive working synergistically with silver diamine fluoride - Google Patents

Abstract

Described herein are functionalized peptides, compositions comprising the same, and methods useful for treatment of dental caries that have been treated with silver diamine fluoride. Treatment with functionalized peptides addresses undesirable tooth discoloration associated with silver diamine fluoride treatment and/or enhances penetration for functional integrity.

Description

PEPTIDE-POLYMER ADHESIVE WORKING SYNERGISTICALLY WITH SILVER DIAMINE FLUORIDE

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] The application claims the benefit of and priority to U.S. Provisional Patent Application No. 63/538723, filed September 15, 2023, which is incorporated by reference herein in its entirety for any and all purposes.

STATEMENT OF U.S. GOVERNMENT SUPPORT

[0002] This invention was made with government support under DE025476 and DE032903 awarded by the National Institutes of Health. The government has certain rights in the invention.

BACKGROUND

[0003] Throughout this disclosure, various publications, patents, and published patent specifications are referenced by an identifying citation. Also within this disclosure are Arabic numerals referring to referenced citations, the full bibliographic details of which are provided subsequent to the Examples section. The disclosures of these publications, patents, and published patent specifications are hereby incorporated by reference into the present disclosure to more fully describe the present technology.

[0004] No infectious disease is more common than dental caries (also referred to by the general population as cavities). A factor in its prevalence is the maternal transmission of cariogenic microbiota. Early childhood caries (ECC) is widely recognized as a global health crisis, with dental caries still the most prevalent chronic disease in children worldwide. Impacting young children under age five, ECC is characterized by the presence of one or more carious lesions of the primary teeth. ECC has a higher prevalence in children of lower socioeconomic groups with limited access to dental care. In the United States, the prevalence of ECC is estimated to be between 3% and 6% of all pre-school aged children, a value which is consistent with literature reviews confirming rates from 1% to 12% in the most developed countries worldwide. However, the worse health outcomes are disproportionately skewed to certain populations as the risk of ECC in disadvantaged populations and in less developed countries can be as high as 70%. Once a child develops carious lesions, the disease becomes more difficult and more expensive to control - rapid disease progression is common without immediate professional intervention. As the ECC disease progresses, treatment options diminish and are often very costly. Children with severe early childhood caries must commonly be treated under general anesthesia. Due to the high cost and potential comorbidities, general anesthesia may not be an option for all children. More devastating to the development of the dentition and growth of the jaws is that the current standard of care in advanced ECC cases recommends premature extraction or extensive dental restorations. Early extraction of the primary teeth can alter jaw growth, leading to the failure of the remaining and adult replacement teeth to work together effectively during chewing. This can lead to consequential changes in micro- and macro-nutrition that adversely impact child health across a lifetime. Therefore, alternative treatment options that circumvent the cascade of failure described above have been the focus of attention in the dental public health community.

SUMMARY

[0005] Caries is the most ubiquitous infectious disease of humankind and early childhood caries (ECC) is the most prevalent chronic disease in children worldwide, with the resulting destruction of the teeth recognized as a global health crisis. Recent Federal Drug Administration (FDA) approval for the use of silver diamine fluoride (SDF) in dentistry offers a safe, accessible, and inexpensive approach to arrest caries progression in children with ECC. However, discoloration, z.e., black staining, of demineralized or cavitated surfaces treated with SDF has limited its widespread use. Furthermore, SDF treatment reduces effective bonding of adhesive dental composite materials commonly used to mask the staining and restore the function of the carious teeth. Therefore, there is a need for compositions and strategies to mitigate the black staining associated with SDF treatment and remineralize carious regions treated with SDF. The present technology addresses this need.

[0006] In an aspect, a functionalized peptide of Formula I is provided:

or a pharmaceutically acceptable salt thereof and/or a solvate thereof, wherein R¹ is hydrogen, unsubstituted Ci-Ce alkyl, or cyano; and Y¹ is a terminal nitrogen, side chain nitrogen, or side chain sulfur of amino acid sequence X1-X2-EQLGVRKELRGV (SEQ ID NO: 1); where Xi is absent or amino acid K, S, R, or E; and X2 is absent or a spacer of 1, 2, 3, 4, 5 ,6 , 7, 8, 9, or 10 amino acid residues.

[0007] In an aspect, a composition is provided that includes the functionalized peptide of the present technology, a dental adhesive, a photoinitiator, and a pharmaceutically acceptable carrier.

[0008] In an aspect, a method of treating a dental surface is provided that includes administering silver diamine fluoride (SDF) to the dental surface; and, after administering the SDF, administering an effective amount of the functionalized peptide of the present technology to the dental surface.

[0009] In an aspect, a method of treating a dental surface is provided that includes administering silver diamine fluoride (SDF) to the dental surface; and, after administering the SDF, administering an effective amount of the composition of the present technology to the dental surface.

BRIEF DESCRIPTION OF THE DRAWINGS

[0010] FIG. 1 provides an illustrative schematic of a method of treating a dental surface with silver diamine fluoride (SDF) and the functionalized peptide of the present technology.

[0011] FIGS. 2A-2D are graphs of attenuated total reflectance-Fourier transform infrared (ATR-FTIR) spectra of methacrylate-functionalized silver binding peptide (MA-AgBP) with or without SDF, illustrating the interaction of 4 mg/mL SDF and 4 mg/mL MA-AgBP (stoichiometry is about 13 SDF complexes to about 1 peptide). FIG. 2A is a graph of the spectra in the region 4000 cm’¹ to 550 cm’¹ of SDF, MA-AgBP, and MA-AgBP with SDF (MA-AgBP/SDF). FIG. 2B is a graph of the spectra in FIG. 2A highlighting the spectral regions corresponding to silver oxides, noted by resolved spectral features at 769 cm’¹ and 610 cm’¹. Without MA-AgBP, the silver in SDF was quickly exposed and oxidized to Ag2O, which overwhelmed the low wavenumber region (SDF spectrum) leading to raised absorbance baseline indicating substantial darkening. With MA-AgBP, Ag⁺ was chelated, the oxidation process was slowed down, silver oxides were formed in a more controlled manner as noted by resolved spectral features at 769 cm'¹ and 610 cm'¹. The Ag-MA-AgBP chelates are relatively transparent, as noted by the relatively flat baseline in this region. FIG. 2C is a graph of the spectra in FIG. 2A highlighting the spectral regions assigned to ammonium fluoride at 2002 cm'¹ and 2261 cm'¹. Diamine-silver ion complexes, [Ag(NHs)2]⁺ were partially replaced by chelated Ag-MA-AgBP. Chelation between Ag and MA-AgBP may prevail over the diammine- Ag ion complexes. Then available ammonia (NH3) molecules may become ionized and form ionic bonds with free fluoride ions from SDF, e.g., (NH₄ ⁺F'). FIG. 2D is a graph of the spectra in FIG. 2A highlighting the spectral regions associated with the ammonium ion (vl and v3 NH₄ modes) at 3088 cm'¹ and 2804 cm'¹.

[0012] FIG. 3 is a bar graph comparing mean fluorescence intensities of SDF-treated dentin, SDF and biotinylated AgBP-treated dentin, and SDF and biotinylated KGSGGG-AgBP- treated dentin after further treatment with a streptavidin-conjugated Q-dot solution. (Oneway Anova, *** denotes p<0.001 (n=8, ±SEM)). This study indicated the similar binding of the AgBP peptide with or without a spacer amino acid sequence KGSGGG to SDF-treated dentinal surfaces.

[0013] FIG. 4 is a bar graph of the average depth of penetration of silver ions into dental samples as measured using micro-CT images of dentin treated with SDF alone or with AgBP peptide, (t-test, n=60, ±SD, ***denotes p<0.001). This study indicated that AgBP may effectively regulate the formation of silver compounds, providing a higher concentration of and deeper penetration of silver ions into the dentin structure.

DETAILED DESCRIPTION

[0014] The following terms are used throughout as defined below.

[0015] As used herein and in the appended claims, singular articles such as “a” and “an” and “the” and similar referents in the context of describing the elements (especially in the context of the following claims) are to be construed to cover both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context. Recitation of ranges of values herein are merely intended to serve as a shorthand method of referring individually to each separate value falling within the range, unless otherwise indicated herein, and each separate value is incorporated into the specification as if it were individually recited herein. All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g., “such as”) provided herein, is intended merely to better illuminate the embodiments and does not pose a limitation on the scope of the claims unless otherwise stated. No language in the specification should be construed as indicating any non-claimed element as essential.

[0016] As used herein, “about” will be understood by persons of ordinary skill in the art and will vary to some extent depending upon the context in which it is used. If there are uses of the term which are not clear to persons of ordinary skill in the art, given the context in which it is used, “about” will mean up to plus or minus 10% of the particular term - for example, “about 10 wt.%” would be understood to mean “9 wt.% to 11 wt.%.” It is to be understood that when “about” precedes a term, the term is to be construed as disclosing “about” the term as well as the term without modification by “about” - for example, “about 10 wt.%” discloses “9 wt.% to 11 wt.%” as well as disclosing “10 wt.%.”

[0017] The phrase “and/or” as used in the present disclosure will be understood to mean any one of the recited members individually or a combination of any two or more thereof - for example, “A, B, and/or C” would mean “A, B, C; A and B; A and C; B and C, or the combination of A, B, and C.”

[0018] As will be understood by one skilled in the art, for any and all purposes, particularly in terms of providing a written description, all ranges disclosed herein also encompass any and all possible subranges and combinations of subranges thereof. Any listed range can be easily recognized as sufficiently describing and enabling the same range being broken down into at least equal halves, thirds, quarters, fifths, tenths, etc. As a non-limiting example, each range discussed herein can be readily broken down into a lower third, middle third and upper third, etc. As will also be understood by one skilled in the art all language such as “up to,” “at least,” “greater than,” “less than,” and the like include the number recited and refer to ranges which can be subsequently broken down into subranges as discussed above. Finally, as will be understood by one skilled in the art, a range includes each individual member. Thus, for example, a group having 1-3 atoms refers to groups having 1, 2, or 3 atoms. Similarly, a group having 1-5 atoms refers to groups having 1, 2, 3, 4, or 5 atoms, and so forth.

[0019] As used herein, the term “peptide” refers to a polymer of amino acid residues joined by amide linkages, which may optionally be chemically modified to achieve desired characteri sties. The term “amino acid residue,” includes but is not limited to amino acid residues contained in the group consisting of alanine (Ala or A), cysteine (Cys or C), aspartic acid (Asp or D), glutamic acid (Glu or E), phenylalanine (Phe or F), glycine (Gly or G), histidine (His or H), isoleucine (He or I), lysine (Lys or K), leucine (Leu or L), methionine (Met or M), asparagine (Asn or N), proline (Pro or P), glutamine (Gin or Q), arginine (Arg or R), serine (Ser or S), threonine (Thr or T), valine (Vai or V), tryptophan (Trp or W), and tyrosine (Tyr or Y) residues. The term “amino acid residue” also may include unnatural amino acids or residues contained in the group consisting of homocysteine, 2-Aminoadipic acid, N-Ethylasparagine, 3 -Aminoadipic acid, Hydroxylysine, P-alanine, P-Amino-propionic acid, allo-Hydroxylysine acid, 2- Aminobutyric acid, 3-Hydroxyproline, 4- Aminobutyric acid, 4-Hydroxyproline, piperidinic acid, 6-Aminocaproic acid, Isodesmosine, 2-Aminoheptanoic acid, allo-Isoleucine, 2-Aminoisobutyric acid, N-Methylglycine, sarcosine, 3- Aminoisobutyric acid, N-Methylisoleucine, 2-Aminopimelic acid, 6-N-Methyllysine, 2,4- Diaminobutyric acid, N-Methylvaline, Desmosine, Norvaline, 2,2'-Diaminopimelic acid, Norleucine, 2,3-Diaminopropionic acid, Ornithine, and N-Ethylglycine. Typically, the amide linkages of the peptides are formed from an amino group of the backbone of one amino acid and a carboxyl group of the backbone of another amino acid.

[0020] By “pharmaceutically acceptable” is meant a material that is not biologically or otherwise undesirable, e.g., the material may be incorporated into a pharmaceutical composition administered to a patient without causing any undesirable biological effects or interacting in a deleterious manner with any of the other components of the composition in which it is contained. When the term “pharmaceutically acceptable” is used to refer to a pharmaceutical carrier or excipient, it is implied that the carrier or excipient has met the required standards of toxicological and manufacturing testing or that it is included on the Inactive Ingredient Guide prepared by the U.S. Food and Drug administration.

[0021] Pharmaceutically acceptable salts of peptides described herein are within the scope of the present technology and include acid or base addition salts which retain the desired pharmacological activity and is not biologically undesirable (e.g., the salt is not unduly toxic, allergenic, or irritating, and is bioavailable). When the compound of the present technology has a basic group, such as, for example, an amino group, pharmaceutically acceptable salts can be formed with inorganic acids (such as hydrochloric acid, hydroboric acid, nitric acid, sulfuric acid, and phosphoric acid), organic acids (e.g., alginate, formic acid, acetic acid, benzoic acid, gluconic acid, fumaric acid, oxalic acid, tartaric acid, lactic acid, maleic acid, citric acid, succinic acid, malic acid, methanesulfonic acid, benzenesulfonic acid, naphthalene sulfonic acid, and p-toluenesulfonic acid) or acidic amino acids (such as aspartic acid and glutamic acid). When the compound of the present technology has an acidic group, such as for example, a carboxylic acid group, it can form salts with metals, such as alkali and earth alkali metals (e.g., Na+, Li+, K+, Ca2+, Mg2+, Zn2+), ammonia or organic amines (e.g., dicyclohexylamine, trimethylamine, triethylamine, pyridine, picoline, ethanolamine, diethanolamine, triethanolamine) or basic amino acids (e.g., arginine, lysine and ornithine). Such salts can be prepared in situ during isolation and purification of the compounds or by separately reacting the purified compound in its free base or free acid form with a suitable acid or base, respectively, and isolating the salt thus formed.

[0022] The peptides of the present technology may exist as solvates, especially hydrates. Hydrates may form during manufacture of the compounds or compositions comprising the compounds, or hydrates may form over time due to the hygroscopic nature of the compounds. Compounds of the present technology may exist as organic solvates as well, including DMF, ether, and alcohol solvates, among others. The identification and preparation of any particular solvate is within the skill of the ordinary artisan of synthetic organic or medicinal chemistry.

[0023] As used herein, “subject” refers to an animal, such as a mammal (including a human), that has been or will be the object of treatment, observation, or experiment. “Subject” and “patient” may be used interchangeably, unless otherwise indicated. Mammals include, but are not limited to, mice, rodents, rats, simians, humans, farm animals, dogs, cats, sport animals, and pets. The methods described herein may be useful in human therapy and/or veterinary applications. In some embodiments, the subject is a mammal. In some embodiments, the subject is a human.

[0024] The term “treatment” or “treating” means administering a compound disclosed herein for the purpose of: (i) delaying the onset of a disease, that is, causing the clinical symptoms of the disease not to develop or delaying the development thereof; (ii) inhibiting the disease, that is, arresting the development of clinical symptoms; (iii) relieving the disease, that is, causing the regression of clinical symptoms or the severity thereof; and/or (iv) alleviating or reducing side-effects of another treatment. [0025] The term “dental surface” refers to a surface of a tooth made of hard tissue that can be treated with the peptides of the present technology. The hard tissue may include enamel, dentin, and/or cementum.

[0026] Generally, reference to a certain element such as hydrogen or H is meant to include all isotopes of that element. For example, if an R group is defined to include hydrogen or H, it also includes deuterium and tritium. Compounds comprising radioisotopes such as tritium, ¹⁴C, ³²P, and ³⁵S are thus within the scope of the present technology. Procedures for inserting such labels into the compounds of the present technology will be readily apparent to those skilled in the art based on the disclosure herein.

[0027] In general, “substituted” refers to an organic group as defined below (e.g., an alkyl group) in which one or more bonds to a hydrogen atom contained therein are replaced by a bond to non-hydrogen or non-carbon atoms. Substituted groups also include groups in which one or more bonds to a carbon(s) or hydrogen(s) atom are replaced by one or more bonds, including double or triple bonds, to a heteroatom. Thus, a substituted group is substituted with one or more substituents, unless otherwise specified. In some embodiments, a substituted group is substituted with 1, 2, 3, 4, 5, or 6 substituents. Examples of substituent groups include: halogens (z.e., F, Cl, Br, and I); hydroxyls; alkoxy, alkenoxy, aryloxy, aralkyloxy, heterocyclyl, heterocyclylalkyl, heterocyclyloxy, and heterocyclylalkoxy groups; carbonyls (oxo); carboxylates; esters; urethanes; oximes; hydroxylamines; alkoxyamines; aralkoxyamines; thiols; sulfides; sulfoxides; sulfones; sulfonyls; pentafluorosulfanyl (z.e., SFs), sulfonamides; amines; N-oxides; hydrazines; hydrazides; hydrazones; azides; amides; ureas; amidines; guanidines; enamines; imides; isocyanates; isothiocyanates; cyanates; thiocyanates; imines; nitro groups; nitriles (z.e., CN); and the like.

[0028] Substituted ring groups such as substituted cycloalkyl, aryl, heterocyclyl and heteroaryl groups also include rings and ring systems in which a bond to a hydrogen atom is replaced with a bond to a carbon atom. Therefore, substituted cycloalkyl, aryl, heterocyclyl and heteroaryl groups may also be substituted with substituted or unsubstituted alkyl, alkenyl, and alkynyl groups as defined below.

[0029] Alkyl groups include straight chain and branched chain alkyl groups having from 1 to 12 carbon atoms, and typically from 1 to 10 carbons or, in some embodiments, from 1 to 8, 1 to 6, or 1 to 4 carbon atoms. Alkyl groups may be substituted or unsubstituted. Examples of straight chain alkyl groups include groups such as methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, and n-octyl groups. Examples of branched alkyl groups include, but are not limited to, isopropyl, iso-butyl, sec-butyl, tert-butyl, neopentyl, isopentyl, and 2,2- dimethylpropyl groups. Representative substituted alkyl groups may be substituted one or more times with substituents such as those listed above, and include without limitation haloalkyl (e.g., trifluoromethyl), hydroxyalkyl, thioalkyl, aminoalkyl, alkylaminoalkyl, dialkylaminoalkyl, alkoxyalkyl, carboxyalkyl, and the like.

[0030] Cycloalkyl groups include mono-, bi- or tricyclic alkyl groups having from 3 to 12 carbon atoms in the ring(s), or, in some embodiments, 3 to 10, 3 to 8, or 3 to 4, 5, or 6 carbon atoms. Cycloalkyl groups may be substituted or unsubstituted. Exemplary monocyclic cycloalkyl groups include, but not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl groups. In some embodiments, the cycloalkyl group has 3 to 8 ring members, whereas in other embodiments the number of ring carbon atoms range from 3 to 5, 3 to 6, or 3 to 7. Bi- and tricyclic ring systems include both bridged cycloalkyl groups and fused rings, such as, but not limited to, bicyclo[2.1.1]hexane, adamantyl, decalinyl, and the like. Substituted cycloalkyl groups may be substituted one or more times with, non-hydrogen and non-carbon groups as defined above. However, substituted cycloalkyl groups also include rings that are substituted with straight or branched chain alkyl groups as defined above. Representative substituted cycloalkyl groups may be mono-substituted or substituted more than once, such as, but not limited to, 2,2-, 2,3-, 2,4- 2,5- or 2,6-disubstituted cyclohexyl groups, which may be substituted with substituents such as those listed above.

[0031] Cycloalkylalkyl groups are alkyl groups as defined above in which a hydrogen or carbon bond of an alkyl group is replaced with a bond to a cycloalkyl group as defined above. Cycloalkylalkyl groups may be substituted or unsubstituted. In some embodiments, cycloalkylalkyl groups have from 4 to 16 carbon atoms, 4 to 12 carbon atoms, and typically 4 to 10 carbon atoms. Substituted cycloalkylalkyl groups may be substituted at the alkyl, the cycloalkyl or both the alkyl and cycloalkyl portions of the group. Representative substituted cycloalkylalkyl groups may be mono-substituted or substituted more than once, such as, but not limited to, mono-, di- or tri -substituted with substituents such as those listed above.

[0032] Alkenyl groups include straight and branched chain alkyl groups as defined above, except that at least one double bond exists between two carbon atoms. Alkenyl groups may be substituted or unsubstituted. Alkenyl groups have from 2 to 12 carbon atoms, and typically from 2 to 10 carbons or, in some embodiments, from 2 to 8, 2 to 6, or 2 to 4 carbon atoms. In some embodiments, the alkenyl group has one, two, or three carbon-carbon double bonds. Examples include, but are not limited to vinyl, allyl, -CH=CH(CH3), -CH=C(CH₃)₂, -C(CH₃)=CH₂, -C(CH₃)=CH(CH₃), -C(CH₂CH₃)=CH₂, among others. Representative substituted alkenyl groups may be mono-substituted or substituted more than once, such as, but not limited to, mono-, di- or tri-substituted with substituents such as those listed above.

[0033] Cycloalkenyl groups include cycloalkyl groups as defined above, having at least one double bond between two carbon atoms. Cycloalkenyl groups may be substituted or unsubstituted. In some embodiments the cycloalkenyl group may have one, two or three double bonds but does not include aromatic compounds. Cycloalkenyl groups have from 4 to 14 carbon atoms, or, in some embodiments, 5 to 14 carbon atoms, 5 to 10 carbon atoms, or even 5, 6, 7, or 8 carbon atoms. Examples of cycloalkenyl groups include cyclohexenyl, cyclopentenyl, cyclohexadienyl, cyclobutadienyl, and cyclopentadienyl.

[0034] Cycloalkenylalkyl groups are alkyl groups as defined above in which a hydrogen or carbon bond of the alkyl group is replaced with a bond to a cycloalkenyl group as defined above. Cycloalkenylalkyl groups may be substituted or unsubstituted. Substituted cycloalkenylalkyl groups may be substituted at the alkyl, the cycloalkenyl or both the alkyl and cycloalkenyl portions of the group. Representative substituted cycloalkenylalkyl groups may be substituted one or more times with substituents such as those listed above.

[0035] Alkynyl groups include straight and branched chain alkyl groups as defined above, except that at least one triple bond exists between two carbon atoms. Alkynyl groups may be substituted or unsubstituted. Alkynyl groups have from 2 to 12 carbon atoms, and typically from 2 to 10 carbons or, in some embodiments, from 2 to 8, 2 to 6, or 2 to 4 carbon atoms. In some embodiments, the alkynyl group has one, two, or three carbon-carbon triple bonds. Examples include, but are not limited to -C=CH, -C=CCH₃, -CH₂C=CCH₃, -C=CCH₂CH(CH₂CH₃)₂, among others. Representative substituted alkynyl groups may be mono-substituted or substituted more than once, such as, but not limited to, mono-, di- or trisubstituted with substituents such as those listed above.

[0036] Aryl groups are cyclic aromatic hydrocarbons that do not contain heteroatoms. Aryl groups may be substituted or unsubstituted. Aryl groups herein include monocyclic, bicyclic, and tricyclic ring systems. Thus, aryl groups include, but are not limited to, phenyl, azulenyl, heptalenyl, biphenyl, fluorenyl, phenanthrenyl, anthracenyl, indenyl, indanyl, pentalenyl, and naphthyl groups. In some embodiments, aryl groups contain 6-14 carbons, and in others from 6 to 12 or even 6-10 carbon atoms in the ring portions of the groups. In some embodiments, the aryl groups are phenyl or naphthyl. The phrase “aryl groups” includes groups containing fused rings, such as fused aromatic-aliphatic ring systems (e.g., indanyl, tetrahydronaphthyl, and the like). Representative substituted aryl groups may be mono-substituted or substituted more than once. For example, monosubstituted aryl groups include, but are not limited to, 2-, 3-, 4-, 5-, or 6-substituted phenyl or naphthyl groups, which may be substituted with substituents such as those listed above.

[0037] Aralkyl groups are alkyl groups as defined above in which a hydrogen or carbon bond of an alkyl group is replaced with a bond to an aryl group as defined above. Aralkyl groups may be substituted or unsubstituted. In some embodiments, aralkyl groups contain 7 to 16 carbon atoms, 7 to 14 carbon atoms, or 7 to 10 carbon atoms. Substituted aralkyl groups may be substituted at the alkyl, the aryl or both the alkyl and aryl portions of the group. Representative aralkyl groups include but are not limited to benzyl and phenethyl groups and fused (cycloalkylaryl)alkyl groups such as 4-indanylethyl. Representative substituted aralkyl groups may be substituted one or more times with substituents such as those listed above.

[0038] Heterocyclyl groups include aromatic (also referred to as heteroaryl) and nonaromatic ring compounds containing 3 or more ring members, of which one or more is a heteroatom such as, but not limited to, N, O, and S. Heterocyclyl groups may be substituted or unsubstituted. In some embodiments, the heterocyclyl group contains 1, 2, 3 or 4 heteroatoms. In some embodiments, heterocyclyl groups include mono-, bi- and tricyclic rings having 3 to 16 ring members, whereas other such groups have 3 to 6, 3 to 10, 3 to 12, or 3 to 14 ring members. Heterocyclyl groups encompass aromatic, partially unsaturated and saturated ring systems, such as, for example, imidazolyl, imidazolinyl and imidazolidinyl groups. The phrase “heterocyclyl group” includes fused ring species including those comprising fused aromatic and non-aromatic groups, such as, for example, benzotri azolyl, 2,3-dihydrobenzo[l,4]dioxinyl, and benzo[l,3]dioxolyl. The phrase also includes bridged polycyclic ring systems containing a heteroatom such as, but not limited to, quinuclidyl. Heterocyclyl groups include, but are not limited to, aziridinyl, azetidinyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, thiazolidinyl, tetrahydrothiophenyl, tetrahydrofuranyl, dioxolyl, furanyl, thiophenyl, pyrrolyl, pyrrolinyl, imidazolyl, imidazolinyl, pyrazolyl, pyrazolinyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, thiazolyl, thiazolinyl, isothiazolyl, thiadiazolyl, oxadiazolyl, piperidyl, piperazinyl, morpholinyl, thiomorpholinyl, tetrahydropyranyl, tetrahydrothiopyranyl, oxathiane, dioxyl, dithianyl, pyranyl, pyridyl, pyrimidinyl, pyridazinyl, pyrazinyl, triazinyl, dihydropyridyl, dihydrodithiinyl, dihydrodithionyl, homopiperazinyl, quinuclidyl, indolyl, indolinyl, isoindolyl,azaindolyl (pyrrolopyridyl), indazolyl, indolizinyl, benzotriazolyl, benzimidazolyl, benzofuranyl, benzothiophenyl, benzthiazolyl, benzoxadiazolyl, benzoxazinyl, benzodithiinyl, benzoxathiinyl, benzothiazinyl, benzoxazolyl, benzothiazolyl, benzothiadiazolyl, benzo [1,3] dioxolyl, pyrazolopyridyl, imidazopyridyl (azabenzimidazolyl), triazolopyridyl, isoxazolopyridyl, purinyl, xanthinyl, adeninyl, guaninyl, quinolinyl, isoquinolinyl, quinolizinyl, quinoxalinyl, quinazolinyl, cinnolinyl, phthalazinyl, naphthyridinyl, pteridinyl, thianaphthyl, dihydrobenzothiazinyl, dihydrobenzofuranyl, dihydroindolyl, dihydrobenzodioxinyl, tetrahydroindolyl, tetrahydroindazolyl, tetrahydrobenzimidazolyl, tetrahydrobenzotriazolyl, tetrahydropyrrolopyridyl, tetrahydropyrazolopyridyl, tetrahydroimidazopyridyl, tetrahydrotriazolopyridyl, and tetrahydroquinolinyl groups. Representative substituted heterocyclyl groups may be mono- substituted or substituted more than once, such as, but not limited to, pyridyl or morpholinyl groups, which are 2-, 3-, 4-, 5-, or 6-substituted, or disubstituted with various substituents such as those listed above.

[0039] Heteroaryl groups are aromatic ring compounds containing 5 or more ring members, of which, one or more is a heteroatom such as, but not limited to, N, O, and S. Heteroaryl groups may be substituted or unsubstituted. Heteroaryl groups include, but are not limited to, groups such as pyrrolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, thiazolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, thiophenyl, benzothiophenyl, furanyl, benzofuranyl, indolyl, azaindolyl (pyrrolopyridinyl), indazolyl, benzimidazolyl, imidazopyridinyl (azabenzimidazolyl), pyrazolopyridinyl, triazolopyridinyl, benzotriazolyl, benzoxazolyl, benzothiazolyl, benzothiadiazolyl, imidazopyridinyl, isoxazolopyridinyl, thianaphthyl, purinyl, xanthinyl, adeninyl, guaninyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl, quinoxalinyl, and quinazolinyl groups. Heteroaryl groups include fused ring compounds in which all rings are aromatic such as indolyl groups and include fused ring compounds in which only one of the rings is aromatic, such as 2,3-dihydro indolyl groups. The phrase “heteroaryl groups” includes fused ring compounds. Representative substituted heteroaryl groups may be substituted one or more times with various substituents such as those listed above.

[0040] Heterocyclylalkyl groups are alkyl groups as defined above in which a hydrogen or carbon bond of an alkyl group is replaced with a bond to a heterocyclyl group as defined above. Heterocyclylalkyl groups may be substituted or un substituted. Substituted heterocyclylalkyl groups may be substituted at the alkyl, the heterocyclyl or both the alkyl and heterocyclyl portions of the group. Representative heterocyclyl alkyl groups include, but are not limited to, morpholin-4-yl-ethyl, furan-2-yl-methyl, imidazol-4-yl-methyl, pyri din-3 - yl-methyl, tetrahydrofuran-2-yl-ethyl, and indol-2-yl-propyl. Representative substituted heterocyclylalkyl groups may be substituted one or more times with substituents such as those listed above.

[0041] Heteroaralkyl groups are alkyl groups as defined above in which a hydrogen or carbon bond of an alkyl group is replaced with a bond to a heteroaryl group as defined above. Heteroaralkyl groups may be substituted or unsubstituted. Substituted heteroaralkyl groups may be substituted at the alkyl, the heteroaryl or both the alkyl and heteroaryl portions of the group. Representative substituted heteroaralkyl groups may be substituted one or more times with substituents such as those listed above.

[0042] Groups described herein having two or more points of attachment (i.e., divalent, trivalent, or polyvalent) within the compound of the present technology are designated by use of the suffix, “ene.” For example, divalent alkyl groups are alkylene groups, divalent aryl groups are arylene groups, divalent heteroaryl groups are divalent heteroarylene groups, and so forth. Substituted groups having a single point of attachment to the compound of the present technology are not referred to using the “ene” designation. Thus, e.g., chloroethyl is not referred to herein as chloroethylene.

[0043] Alkoxy groups are hydroxyl groups (-OH) in which the bond to the hydrogen atom is replaced by a bond to a carbon atom of a substituted or unsubstituted alkyl group as defined above. Alkoxy groups may be substituted or unsubstituted. Examples of linear alkoxy groups include but are not limited to methoxy, ethoxy, propoxy, butoxy, pentoxy, hexoxy, and the like. Examples of branched alkoxy groups include but are not limited to isopropoxy, sec-butoxy, tert-butoxy, isopentoxy, isohexoxy, and the like. Examples of cycloalkoxy groups include but are not limited to cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy, and the like. Representative substituted alkoxy groups may be substituted one or more times with substituents such as those listed above.

[0044] The terms “alkanoyl” and “alkanoyloxy” as used herein can refer, respectively, to - C(O)-alkyl groups and -O-C(O)-alkyl groups, each containing 2-5 carbon atoms. Similarly, “aryloyl” and “aryloyloxy” refer to -C(O)-aryl groups and -O-C(O)-aryl groups.

[0045] The terms "aryloxy" and “arylalkoxy” refer to, respectively, a substituted or unsubstituted aryl group bonded to an oxygen atom and a substituted or unsubstituted aralkyl group bonded to the oxygen atom at the alkyl. Examples include but are not limited to phenoxy, naphthyloxy, and benzyloxy. Representative substituted aryloxy and arylalkoxy groups may be substituted one or more times with substituents such as those listed above.

[0046] The term “urea” refers to -NR⁸⁴-C(O)-NR⁸⁵R⁸⁶ groups. R⁸⁴, R⁸⁵, and R⁸⁶ groups are independently hydrogen, or a substituted or unsubstituted alkyl, alkenyl, alkynyl, cycloalkyl, aryl, aralkyl, heterocyclyl, or heterocyclylalkyl group as defined herein.

[0047] The term “halogen” or “halo” as used herein refers to bromine, chlorine, fluorine, or iodine. In some embodiments, the halogen is fluorine. In other embodiments, the halogen is chlorine or bromine.

[0048] Those of skill in the art will appreciate that compounds of the present technology may exhibit the phenomena of tautomerism, conformational isomerism, geometric isomerism and/or stereoisomerism. As the formula drawings within the specification and claims can represent only one of the possible tautomeric, conformational isomeric, stereochemical or geometric isomeric forms, it should be understood that the present technology encompasses any tautomeric, conformational isomeric, stereochemical and/or geometric isomeric forms of the compounds having one or more of the utilities described herein, as well as mixtures of these various different forms.

[0049] “ Tautomers” refers to isomeric forms of a compound that are in equilibrium with each other. The presence and concentrations of the isomeric forms will depend on the environment the compound is found in and may be different depending upon, for example, whether the compound is a solid or is in an organic or aqueous solution. For example, in aqueous solution, quinazolinones may exhibit the following isomeric forms, which are referred to as tautomers of each other:

As another example, guanidines may exhibit the following isomeric forms in protic organic solution, also referred to as tautomers of each other:

Because of the limits of representing compounds by structural formulas, it is to be understood that all chemical formulas of the compounds described herein represent all tautomeric forms of compounds and are within the scope of the present technology.

[0050] Stereoisomers of compounds (also known as optical isomers) include all chiral, diastereomeric, and racemic forms of a structure, unless the specific stereochemistry is expressly indicated. Thus, compounds used in the present technology include enriched or resolved optical isomers at any or all asymmetric atoms as are apparent from the depictions. Both racemic and diastereomeric mixtures, as well as the individual optical isomers can be isolated or synthesized so as to be substantially free of their enantiomeric or diastereomeric partners, and these stereoisomers are all within the scope of the present technology.

[0051] The compounds of the present technology may exist as solvates, especially hydrates. Hydrates may form during manufacture of the compounds or compositions comprising the compounds, or hydrates may form over time due to the hygroscopic nature of the compounds. Compounds of the present technology may exist as organic solvates as well, including DMF, ether, and alcohol solvates, among others. The identification and preparation of any particular solvate is within the skill of the ordinary artisan of synthetic organic or medicinal chemistry.

[0052] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this present technology belongs. Although any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the present technology, representative illustrative methods and materials are described herein.

[0053] The Present Technology

[0054] There is a continuing need to improve treatment of dental caries. Fluorides have proven useful to slow the progression of dental caries by replacing the hydroxyl group with fluoride in the hydroxyapatite biomineral of the teeth, thereby inhibiting the carious demineralization of teeth. Recently, silver diamine fluoride has gained attention as a safe, accessible, and inexpensive approach to arrest caries progression in children with ECC. SDF has been used worldwide for decades but was approved for dental use by the FDA in 2014. Although the exact mechanism of action has not been resolved, silver ions deposited on the dental tissues have notable antimicrobial properties. SDF treatment may result in silver ions and silver nanostructures deposited on the dental surface. Recently, silver microwires have been described in teeth treated with SDF, where, without being bound by any theory, the SDF may deposit microwires that replace defects that are a result of caries-provoked demineralization.

[0055] Unlike surgical interventions which require skilled professionals, SDF treatments can be applied by a wider range of health care providers. SDF works by limiting caries progression and protecting teeth from further degradation. As of 2016, the 38% SDF solution was awarded breakthrough therapy status by the US Food and Drug Administration for use in treating ECC. Numerous case studies have shown the overwhelming benefit of single or biannual SDF treatments for caries arrest, with a focus for use of SDF in primary teeth of children affected by ECC. SDF is inexpensive, application takes minutes, and it does not require significant patient cooperation. Only rare minor gingival irritation and no serious adverse events are associated with the use of SDF.

[0056] A side effect of SDF is black staining of the treated demineralized or cavitated surfaces due to the deposition of silver metal and ions. This side effect has limited the widespread adoption of SDF treatment for caries arrest. The loss of aesthetics was documented by Crystal et al., who interviewed parents of children qualified to receive SDF treatment and found that roughly one third of parents found the treatment unacceptable under any circumstances. Crystal, Y.O.; Janal, M.N.; Hamilton, D.S.; Niederman, R., The Journal of the American Dental Association 2017, 148, 510-518, e514. In some trials, mild gingiva irritation and redness were noted, but these symptoms subsided in a few days.

[0057] SDF is not recommended for use on carious lesions that extend into the dental pulp as it will not arrest the progression of the infection. In addition, SDF treatment is recognized to reduce effective bonding of adhesive dental composite materials commonly used to mask the staining and restore the function of the carious teeth.

[0058] Functionalized Peptides

[0059] In an aspect, the technology provides a functionalized peptide to reduce SDF silver oxidation staining and provide a composite bondable interface. The functionalized peptide may work synergistically with SDF to create an integrated interface that inhibits the silver- oxidation process while also being polymerizable to form a peptide-polymer composite at the dental surface. The composite may provide extended residence times of the functionalized peptide at the tooth surface while maintaining the peptide’s bioactivity. The composite may also provide a substrate suitable for bonding with composite-restorative materials.

[0060] The functionalized peptide has the structure of Formula I:

Or one or both of a pharmaceutically acceptable salt thereof and/or a solvate thereof, where R¹ is hydrogen, unsubstituted Ci-Ce alkyl, or cyano; and Y¹ is a terminal nitrogen, side chain nitrogen, or side chain sulfur of amino acid sequence X1-X2-EQLGVRKELRGV (SEQ ID NO: 1); where Xi is absent or amino acid K, S, R, or E; and X2 is absent or a spacer of 1, 2, 3, 4, 5 ,6 , 7, 8, 9, or 10 amino acid residues.

[0061] The functionalized peptide of the present technology may include one or more D- amino acids as well as one or more L-amino acids. In any embodiment herein, the functionalized peptide may consist of only D-amino acids, or alternatively in any embodiment herein the peptide may consist only of L-amino acids. As discussed herein, the functionalized peptide of the present technology consists of a silver binding peptide (AgBP) portion, an optional spacer portion, and an acrylate portion. The functionalized peptide of the present technology achieves 1) targeted surface binding to silver nanostructures on dental surfaces and 2) polymerization to provide extended residence times at the dental surface. The functionalized peptide can self-assemble on complex surfaces having silver ions or nanostructures thereon to produce an interface serving to modulate functions that favorably direct biomineralization.

[0062] A peptide of the present technology may be synthesized by any technique known to those of skill in the art and by methods as disclosed herein. Methods for synthesizing the disclosed peptides may include chemical synthesis of proteins or peptides, the expression of peptides through standard molecular biological techniques, and/or the isolation of proteins or peptides from natural sources. The disclosed peptides thus synthesized may be subject to further chemical and/or enzymatic modification. Various methods for commercial preparations of peptides and polypeptides are known to those of skill in the art.

[0063] A peptide of the present technology may alternatively be made by recombinant means or by cleavage from a longer polypeptide. The composition of a peptide may be confirmed by amino acid analysis or sequencing.

[0064] The peptide of the present technology may be functionalized with an acrylate portion by chemical modification by any technique known to those of skill in the art and by methods as disclosed herein. Methods for functionalizing the peptide with an acrylate portion may include chemical modification by reacting an acrylic molecule with a terminal nitrogen, side chain nitrogen, or side chain sulfur of amino acid sequence to form an acrylamide linkage, where the molecule’s reactive C=C bond is reserved for subsequent participation in the polymerization reaction. See Sheng-Xue Xie, Linyong Song, Esra Yuca, Kyle Boone, Rizacan Sarikaya, Sarah Kay VanOosten, Anil Misra, Qiang Ye, Paulette Spencer, and Candan Tamerler ACS Applied Polymer Materials 2020 2 (3), 1134-1144 DOI: 10.1021/acsapm.9b00921; Yuca, E.; Xie, S.-X.; Song, L.; Boone, K.; Kamathewatta, N.; Woolfolk, S.K.; Elrod, P.; Spencer, P.; Tamerler, C. Reconfigurable Dual Peptide Tethered Polymer System Offers a Synergistic Solution for Next Generation Dental Adhesives. Int. J. Mol. Sci. 2021, 22, 6552. For example, chemical

modification may include reacting an acrylic molecule with a side chain guanido group of arginine, a side chain amino group of lysine, a side chain thiol group of cysteine, or side chain a carboxyl group of glutamic acid to form an acrylamide linkage.

[0065] The functionalized peptide includes a functional domain that may target, bind, and/or self-assemble on silver particles, silver nanoparticles, and/or silver surfaces. The silver- binding functional domain in the functionalized peptides may be used to selectively target tissue treated with SDF. SDF treatment of a dental surface results in silver metal and/or silver ion deposition on the dental surface. The deposited silver from SDF on the dental surface may act as a selective and specific target for the functionalized peptides binding and/or self-assembly.

[0066] The silver-binding functional domain in the functionalized peptide is a silver binding peptide (AgBP) of amino acid sequence EQLGVRKELRGV (SEQ ID NO: 10). The AgBP may self-assemble on silver particles, silver nanoparticles, and/or silver surfaces, including silver compounds deposited on dental surfaces as a result of SDF treatment to anchor to the SDF -treated surface. A construct with the AgBP domain may have an equilibrium dissociation constant with silver of about 1 to about 3 orders of magnitude lower than a similar construct without the AgBP domain.

[0067] In addition to silver-binding, the functionalized peptides may be polymerized to provide a peptide-polymer composite exhibiting antimicrobial and remineralization properties at the dental surface. The composite may provide extended residence times of the functionalized peptide at the tooth surface as compared to administration of the peptide without polymerization, while maintaining the peptide’s bioactivity. The composite may also provide a substrate suitable for bonding with composite-restorative materials. Further the composite may provide antimicrobial and remineralization properties.

[0068] The acrylate portion of the functionalized peptide has the structure of Formula I where R¹ is hydrogen, unsubstituted Ci-Ce alkyl, or cyano. The C=C group of the acrylate portion is a reactive group that is susceptible to polymerization. Examples of the acrylate portion include the structures of Formula II to IX

[0069] The functionalized peptides may or may not have an amino acid sequence between the AgBP and the acrylate acting as a spacer, referred to in Formula I as X2. The spacer may influence domain activity depending on the spacer’s length and flexibility. The spacer sequences may help ensure that the functional domain AgBP and the reactive group of acrylate still substantially maintain their isolated functions, and/or may reduce potential interference between the functional domain and reactive group. Such spacer sequences include, but are not limited to, EAAAK (SEQ ID NO: 2), APA (SEQ ID NO: 3), GGG (SEQ ID NO: 4), PAPAP (SEQ ID NO: 5), or GSGGG (SEQ ID NO: 6).

[0070] The functionalized peptides may or may not have an additional amino acid between the AgBP and the acrylate acting as a conjugation point, referred to in Formula I as Xi. The additional amino may provide a side chain with a functional group that may react with an acrylate group to form the acrylate portion of the functionalized peptide. Such additional amino acids include, but are not limited to, K, S, C, and E.

[0071] Examples of functionalized peptides according to the present technology include EAAAKEQLGVRKELRGV (SEQ ID NO: 9), EQLGVRKELRGV (SEQ ID NO: 10), APAKEQLGVRKELRGV (SEQ ID NO: 11), GGGKEQLGVRKELRGV (SEQ ID NO: 12), PAPAPKEQLGVRKELRGV (SEQ ID NO: 13), and GS GGGKEQLGVRKELRGV (SEQ ID NO: 14).

[0072] The functionalized peptides are engineered peptides that target and bind to SDF- treated dental surfaces and polymerize to form peptide-polymer composites. Binding to SDF -treated dental surfaces may slow the silver oxidation process, thereby mitigating the black staining resulting from SDF treatment. The composites may provide extended residence times of the functionalized peptide at the tooth surface, as compared to unfunctionalized peptide, while maintaining similar bioactivity. The composites may be adhesive dental composites used to fill in holes and cover SDF staining, and/or the composites may act as stable interfaces for application of an additional adhesive dental composite, where the dental composite may be used to fill in holes and cover SDF staining. Further the composite may provide antimicrobial and remineralization properties. This aspect of acting as an adhesive dental composite is beneficial since a dental composite does not typically adhere well to SDF-treated dental surfaces. The resulting functionalized peptides demonstrate both polymerization and silver-binding functions.

[0073] Compositions

In an aspect, a composition is provided that includes a functionalized peptide of any embodiment disclosed herein, a pharmaceutically acceptable carrier or one or more excipients, fillers, or agents (collectively referred to hereafter as “pharmaceutically acceptable carrier” unless otherwise indicated and/or specified). In a related aspect, a dental adhesive composition for treating dental caries or hypomineralized dental surfaces, and/or for bonding dental restorations to the tooth is provided that includes a functionalized peptide of any embodiment disclosed herein, a photoinitiator, optionally one or more dental adhesives, and optionally a pharmaceutically acceptable carrier. In a related aspect, a medicament for treating dental caries or hypomineralized dental surfaces is provided that includes a functionalized peptide of any embodiment disclosed herein and optionally a pharmaceutically acceptable carrier. In a related aspect, a medicament for controlling biomineralization on a dental surface is provided that includes a functionalized peptide of any embodiment disclosed herein and optionally a pharmaceutically acceptable carrier. In a related aspect, a pharmaceutical composition is provided that includes an effective amount of a functionalized peptide of any embodiment disclosed herein as well as a pharmaceutically acceptable carrier. For ease of reference, the compositions, adhesives, medicaments, and pharmaceutical compositions of the present technology may collectively be referred to herein as “compositions.” In further related aspects, the present technology provides methods and uses that include a functionalized peptide of any aspect or embodiment disclosed herein and/or a composition of any embodiment disclosed herein as well as uses thereof.

[0074] “Effective amount” refers to the amount of a compound, functionalized peptide, or composition required to produce a desired effect. One example of an effective amount includes amounts or dosages that yield acceptable toxicity and bioavailability levels for therapeutic (pharmaceutical) use including, but not limited to facilitating the formation of a polymer on dental surfaces, masking and/or reducing the staining caused by SDF, improving adhesion between a dental adhesive composite and an SDF-treated dental surface, inhibiting caries progression, protecting against future caries formation, and/or rebuilding damaged dental tissue. In any aspect or embodiment disclosed herein (collectively referred to herein as “any embodiment herein,” “any embodiment disclosed herein,” or the like) of the compositions, pharmaceutical compositions, and methods including a functionalized peptide of the present technology, the effective amount may be an amount effective in treatment, including masking and/or reducing the staining caused by SDF, improving adhesion between a dental adhesive composite and an SDF-treated dental surface, inhibiting caries progression, protecting against future caries formation, and/or rebuilding damaged dental tissue. By way of example, the effective amount of any embodiment herein including a functionalized peptide of the present technology may be from about 0.01 pg to about 200 mg of the functionalized peptide (such as from about 0.1 pg to about 50 mg of the functionalized peptide or about 10 pg to about 20 mg of the peptide). The effective amount may be related to the corresponding area and the molecular mass of the functionalized peptide required to saturate an SDF -treated dental surface. The molecular mass required to deliver the corresponding surface coverage could be obtained by converting the number of functionalized peptides that is calculated from the theoretical “footprint” for each functionalized peptide using the variety of peptide structural analyses tools including UCSF Chimera tool. See E. Cate Wisdom, Yan Zhou, Casey Chen, Candan Tamerler, and Malcolm L. Snead, Mitigation of Peri-implantitis by Rational Design of Bifunctional Peptides with Antimicrobial Properties, ACS Biomaterials Science & Engineering 2020 6 (5), 2682-2695 DOI: 10.1021/acsbiomaterials.9b01213. The “theoretical footprint” of the functionalized peptides could be determined through the length and width distance values measured from the a-carbon of amino acid residues. The number of functionalized peptides could be next converted to a molecular mass required to deliver the corresponding dental surface coverage. The methods and uses according to the present technology may include an effective amount of a functionalized peptide of any embodiment disclosed herein. In any aspect or embodiment disclosed herein, the effective amount may be determined in relation to a subject and/or in relation to dental caries. The term “subject” and “patient” may be used interchangeably.

[0075] The functionalized peptide may be incorporated in a dental adhesive composition. The dental adhesive composition may include about 1 wt.% to about 50 wt.% of the functionalized peptide, e.g., about 1 wt.% to about 40 wt.%, about 5 wt.% to about 30 wt.%, about 5 wt.% to about 15 wt.%, or about 10 wt.%. The dental adhesive may be applied to the exposed tooth surface. A dental composite may be placed on top of the adhesive, forming a bond with the adhesive.

[0076] The dental adhesive composition may optionally include a dental adhesive, which may be an additional acrylate monomer or polymer. Acrylate monomers include, but are not limited to, 2,2-bis[4-(2-hydroxyl-3-methacryloxypropoxy)phenyl] propane; 2-hydroxyethyl methacrylate (HEMA); propane-2,2-diylbis[4,l-phenyleneoxy(2-hydroxypropane-3,l-diyl)] bis(2-methylprop-2-enoate), 2-(methacryloyloxy)ethyl (2-(trimethylammonio)ethyl) phosphate (MPC); urethane dimethacrylate (UDMA); 3 -trimethoxy silyl propyl methacrylate (MPS); (trimethoxysilyl)methyl methacrylate (MMeS); 4,4-diethoxy-9-oxo-3,10-dioxa-8-aza- 4-siladodecan- 12-yl methacrylate; trimethoxy silylpropyl methacrylate; a compound having the structure of Formula X

acrylic acid; methacrylic acid, ethyl acrylate; methyl acrylate; butyl acrylate; 2-ethylhexyl acrylate; hydroxy ethyl acrylate; hydroxypropyl acrylate; N-butyl acrylate; 1,3 -butanediol dimethacrylate; 1,6-hexanediol dimethacrylate, and combinations of any two or more thereof. The dental adhesive may be present in the composition in an amount of about 1 wt.% to about 90 wt.% of the dental adhesive, e.g., about 10 wt.% to about 80 wt.%, about 20 wt.% to about 70 wt.%, about 30 wt.% to about 70 wt.%, about 40 wt.% to about 70 wt.%, about 50 wt.% to about 70 wt.%, or about 55 wt.% to about 65 wt.%.

[0077] The dental adhesive composition may include a photoinitiator to initiate acrylate polymerization. Photoinitiators include, but are not limited to, camphoroquinone, ethyl-4- (dimethylamine) benzoate, diphenyliodonoium hexafluorophyosphate, or a combination of two or more thereof. The photoinitiator may be present in an amount of about 0.1 wt.% to about 10 wt.%, e.g., about 0.5 wt.% to about 5 wt.%, about 1 wt.% to about 3 wt.%, or about 2 wt.%.

[0078] Thus, the instant present technology provides dental adhesives, pharmaceutical compositions and medicaments including a functionalized peptide of any embodiment disclosed herein (or a composition of any embodiment disclosed herein) and a pharmaceutically acceptable carrier. The compositions may be used in the methods and treatments described herein. The dental adhesives and pharmaceutical composition may be packaged in unit dosage form. The unit dosage form is effective in treatment, including masking or reducing the staining caused by SDF, improving adhesion between a dental adhesive composite and an SDF-treated dental surface, inhibiting caries progression, protecting against future caries formation, and/or rebuilding damaged dental tissue. Generally, a unit dosage including a functionalized peptide of the present technology will vary depending on patient considerations. Such considerations include, for example, age, protocol, condition, sex, extent of disease, contraindications, concomitant therapies and the like. Further, a unit dosage including a functionalized peptide of the present technology may vary depending on the size of the carious region and SDF treatment. An exemplary unit dosage based on these considerations may also be adjusted or modified by a physician skilled in the art. Suitable unit dosage forms, include, but are not limited to oral solutions, powders, lozenges, topical varnishes, lipid complexes, liquids, etc.

[0079] The dental adhesives, pharmaceutical compositions and medicaments may be prepared by mixing a functionalized peptide of the present technology with one or more pharmaceutically acceptable carriers, excipients, binders, diluents or the like. Such compositions may be in the form of, for example, powders, syrup, emulsions, suspensions, or solutions. The instant compositions may be formulated for various routes of administration, for example, by intraoral administration or via administration (e.g., application) to a dental surface external to a patient. The following dosage forms are given by way of example and should not be construed as limiting the instant present technology.

[0080] For intraoral administration, powders and suspensions are acceptable as solid dosage forms. These may be prepared, for example, by mixing a functionalized peptide of the present technology with at least one additive such as a starch or other additive. Suitable additives are sucrose, lactose, cellulose sugar, mannitol, maltitol, dextran, starch, agar, alginates, chitins, chitosans, pectins, tragacanth gum, gum arabic, gelatins, collagens, casein, albumin, synthetic or semi -synthetic polymers or glycerides. Optionally, oral dosage forms may contain other ingredients to aid in administration, such as an inactive diluent, or lubricants such as magnesium stearate, or preservatives such as paraben or sorbic acid, or anti-oxidants such as ascorbic acid, tocopherol or cysteine, a disintegrating agent, binders, thickeners, buffers, sweeteners, flavoring agents and/or perfuming agents.

[0081] Liquid dosage forms for oral administration (e.g., intraoral administration) may be in the form of pharmaceutically acceptable emulsions, syrups, suspensions, or solutions, which may contain an inactive diluent, such as water. Pharmaceutical formulations and medicaments may be prepared as liquid suspensions or solutions using a sterile liquid, such as, but not limited to, an oil, water, an alcohol, acetone, N-methyl-2-pyrrolidone, dimethyl sulfoxide, propylene glycol, and combinations of these. Pharmaceutically suitable surfactants, suspending agents, and emulsifying agents may be added for oral administration.

[0082] As noted above, suspensions may include oils. Such oils include, but are not limited to, peanut oil, sesame oil, cottonseed oil, corn oil, and olive oil. Suspension preparation may also contain esters of fatty acids such as ethyl oleate, isopropyl myristate, fatty acid glycerides, and acetylated fatty acid glycerides. Suspension formulations may include alcohols, such as, but not limited to, ethanol, isopropyl alcohol, hexadecyl alcohol, glycerol, and propylene glycol. Ethers, such as but not limited to, poly(ethyleneglycol); petroleum hydrocarbons such as mineral oil and petrolatum; acetone, N-methyl-2-pyrrolidone, dimethyl sulfoxide; propylene glycol; and water may also be used in suspension formulations.

[0083] The adhesives, pharmaceutical compositions, and medicaments in liquid or gel form may have a concentration of a functionalized peptide of the present technology sufficient to provide an effective amount as described above. The concentration of the functionalized peptide of the present technology in the adhesives, pharmaceutical compositions, and medicaments may be about 5 pM to about 500 pM (including about 5 pM, about 10 pM, about 50 pM, about 100 pM, about 200 pM, about 500 pM, or any range including and/or in between any two of these values).

[0084] The dental adhesive, pharmaceutical formulation, and/or medicament may be a powder suitable for reconstitution with an appropriate solution as described above. Examples of these include, but are not limited to, freeze dried, rotary dried or spray dried powders, amorphous powders, granules, precipitates, or particulates. The formulations may optionally contain stabilizers, antimicrobial agents, antioxidants, pH modifiers, surfactants, bioavailability modifiers, and combinations of these. The carriers and stabilizers vary with the requirements of the particular composition, but typically include nonionic surfactants (Tweens, Pluronics, or polyethylene glycol), innocuous proteins like serum albumin, sorbitan esters, oleic acid, lecithin, amino acids such as glycine, buffers, salts, sugars, or sugar alcohols. Powders and sprays may be prepared, for example, with excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and polyamide powder, or mixtures of these substances. Ointments, pastes, creams, and gels may also contain excipients such as animal and vegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc and zinc oxide, or mixtures thereof.

[0085] Besides those representative dosage forms described above, pharmaceutically acceptable excipients and carriers are generally known to those skilled in the art and are thus included in the instant present technology. Such excipients and carriers are described, for example, in “Remingtons Pharmaceutical Sciences” Mack Pub. Co., New Jersey (1991), and “Remington: The Science and Practice of Pharmacy,” 20^th Edition, Editor: Alfonso R Gennaro, Lippincott, Williams & Wilkins, Baltimore (2000), each of which is incorporated herein by reference.

[0086] Methods

[0087] Disclosed herein, in one aspect, is a method of masking or reducing the staining caused by SDF and improving adhesion between a dental adhesive composite and an SDF- treated dental surface by applying a functionalized peptide of the present technology or a composition containing the functionalized peptide to a dental surface, e.g., to produce a film on the dental surface. FIG. 1 provides an illustrative schematic of a method of treating a dental surface with silver diamine fluoride (SDF) and the functionalized peptide of the present technology.

[0088] In another aspect, provided herein are methods of treating dental caries in a subject in need thereof, the methods comprising, consisting essentially of, or consisting of administering an effective amount of a functionalized peptide of the present technology or a composition of the present technology to a dental surface treated with SDF in the subject.

[0089] In any of the methods disclosed herein, SDF may be administered to the dental surface, and, after administering the SDF, an effective amount of a functionalized peptide of the present technology or a composition containing the functionalized peptide may be administered to the dental surface. The dental surface may be a dental enamel and/or dentin. The dental enamel and/or dentin may include a carious region, a hypomineralized region, or both a carious region and a hypomineralized region. Administering SDF to the dental surface may include administering a solution comprising SDF to the dental surface. As an example, the solution may have an SDF concentration of about 38% w/v.

[0090] After administering the functionalized peptide of the present technology or a composition containing the functionalized peptide, the methods may further include exposing the dental surface to light to initiate polymerization of the functionalized peptide and the dental adhesive. The light may have a wavelength of about 200 nm to about 500 nm, e.g., 380 nm to about 500 nm, with a sufficient intensity to initiate polymerization.

[0091] In any of the methods disclosed herein, an effective amount of functionalized peptide or a composition containing the functionalized peptide may be administered to a dental surface for a period of about 10 seconds to about 4 hours, such as for a period of about 10 seconds to about 20 seconds, about 20 seconds to about 1 minute, about 1 minute to about 4 hours, 1 minute to 10 minutes, or 2 hours to 4 hours.

[0092] Treatment with the functionalized peptide may be added to the FDA-approved SDF treatment regime to further increase the arrest of caries progression, mediate remineralization at the SDF-treated dental surface, and mitigate SDF’s adverse effect, z.e., black staining of the treated carious lesion.

[0093] The functionalized peptides may work synergistically with the SDF treatment to help protect and rebuild damaged dental tissues while adding a new mineral layer that may be incorporated in adhesive dental composites to restore function and esthetics to people suffering from dental caries. As discussed previously herein, a functionalized peptide of the present technology may be incorporated in an effective amount into adhesive formulations as part of a peptide-polymer hybrid and/or impregnated into the adhesive. The adhesive is applied to the exposed tooth surface. The dental composite, which is placed on top of the adhesive, forms a bond with the adhesive.

[0094] Few studies have been published to date focusing on masking or reducing the staining of SDF on carious dentin and enamel and have used potassium iodide, composites, or glass ionomer cement. Hamdy et. al. used a calibrated spectrophotometer to monitor changes in tooth color following SDF treatment and found a composite coating was the most successful masking agent at baseline and after an aging protocol of the three treatments. However, the composite treatment that immediately followed SDF application used an etching step and multiple coats of dental composite on the affected surface and the effectiveness of the initial SDF treatment after the composite coverage was not reported. Further, it is known that composite restorations in the oral cavity have a high rate of failure due to cracks and gaps creating spaces for bacterial infiltration leading to further decay.

[0095] A functionalized peptide of the present technology may provide a dental adhesive composite that improves adhesion of the functionalized peptide to the SDF-treated dental surface. The functionalized peptides of the present technology, having AgBP domains for targeting silver, help reinforce interfacial binding between silver on the dental surface and the composite. In this way, the functionalized peptides improve interfacial integrity and binding of polymeric composites to dental surfaces.

[0096] The examples herein are provided to illustrate advantages of the present technology and to further assist a person of ordinary skill in the art with preparing or using the functionalized peptides and compositions of the present technology. The examples herein are also presented in order to more fully illustrate the preferred aspects of the present technology. The examples should in no way be construed as limiting the scope of the present technology, as defined by the appended claims. The examples may include or incorporate any of the variations, aspects, or embodiments of the present technology described above. The variations, aspects, or embodiments described above may also further each include or incorporate the variations of any or all other variations, aspects, or embodiments of the present technology.

EXAMPLES

[0097] Example 1: Methacrylate-Functionalized Silver Binding Peptides

[0098] A methacrylate-functionalized AgBP was synthesized having the structure according to Formula I, where Xi was K and X2 was GSGGG.

[0099] FIGS. 2A-2D are graphs of attenuated total reflectance-Fourier transform infrared (ATR-FTIR) spectra of methacrylate-functionalized silver binding peptide (MA- AgBP) with or without SDF, illustrating the interaction of 4 mg/mL SDF and 4 mg/mL MA- AgBP (stoichiometry is about 13 SDF complexes to about 1 peptide). FIG. 2A is a graph of the spectra in the region 4000 cm’¹ to 550 cm’¹ of SDF, MA-AgBP, and MA-AgBP with SDF (MA-AgBP/SDF).

[0100] FIG. 2B is a graph of the spectra in FIG. 2A highlighting the spectral regions corresponding to silver oxides, noted by resolved spectral features at 769 cm’¹ and 610 cm’¹. Without MA-AgBP, the silver in SDF was quickly exposed and oxidized to Ag2O, which overwhelmed the low wavenumber region (SDF spectrum) leading to raised absorbance baseline indicating substantial darkening. With MA-AgBP, Ag⁺ was chelated, the oxidation process was slowed down, silver oxides were formed in a more controlled manner as noted by resolved spectral features at 769 cm’¹ and 610 cm’¹. The Ag-MA-AgBP chelates are relatively transparent, as noted by the relatively flat baseline in this region.

[0101] FIG. 2C is a graph of the spectra in FIG. 2A highlighting the spectral regions assigned to ammonium fluoride at 2002 cm’¹ and 2261 cm’¹. Diamine-silver ion complexes, [Ag(NH3)2]⁺ were partially replaced by chelated Ag-MA-AgBP. Chelation between Ag and MA-AgBP may prevail over the diammine- Ag ion complexes. Then available ammonia (NH3) molecules may become ionized and form ionic bonds with free fluoride ions from SDF, e g., (NH₄ ⁺F ).

[0102] FIG. 2D is a graph of the spectra in FIG. 2A highlighting the spectral regions associated with the ammonium ion (vl and v3 NH4 modes) at 3088 cm'¹ and 2804 cm'¹.

[0103] Example 2: Application of Biotin-Functionalized AgBP of SDF-Treated Dentin

[0104] Peptide Synthesis. The silver-binding peptides used in this study, AgBP (sequence: EQLGVRKELRGV) and spacer-AgBP (KGSGGG-EQLGVRKELRGV), were synthesized using an Fmoc-based solid-phase peptide synthesis protocol on an AAPPTEC Focus XC synthesizer (KY, USA). Peptides were biotinylated post-synthesis for subsequent fluorescence studies using streptavidin-conjugated Q-dots. The biotinylation protocol followed the AAPPtec standard protocol for the activation and coupling of d-biotin (AAPPTEC, KY, USA) to the peptide N-terminus. After biotinylation, the peptides were washed with dimethylformamide (DMF, 99.8%, Sigma-Aldrich, MO, USA) and 200-proof ethanol and dried under vacuum at room temperature. The peptide was then cleaved from the resin and side-chains deprotected, following the standard AAPPTEC protocol, using a cleavage cocktail containing trifluoroacetic acid (TFA, 99%, Sigma-Aldrich, MO, USA), phenol (89%, Fisher Scientific, NJ, USA), triisopropylsilane (98%, Sigma-Aldrich, MO, USA), and deionized (DI) water at a ratio of 90.0:5.0:2.5:2.5 ( /v percent). Peptide cleavage was run for three hours on a rotator. The peptides were precipitated with cold ether and lyophilized. An analytical Shimadzu HPLC system, LC-2010 HT liquid chromatograph, and SPD-M20A prominence diode array detector were used to confirm peptide purity. A 5 pm C- 18 silica Luna column (250x4.6 mm, Phenomenex Inc., CA, USA) was utilized with a mobile phase consisting of 100% acetonitrile and phase A (99.9% HPLC-grade water, 0.1% TFA). The system was operated at 40 °C, 1 mL/min flow rate, and 254 nm detection on a linear gradient. Lyophilized peptides were stored at -20 °C prior to use in subsequent experiments.

[0105] Dentin Specimen Preparation. This study specifically involved teeth that had already been scheduled for extraction. Following extraction, the teeth were placed in individual vials filled with phosphate-buffered saline solution that had been modified with 20 ppm sodium azide and stored at 4 °C. To ensure that the teeth were representative of the population residing in Kansas City and its surrounding areas, they were randomly selected from a diverse group of male and female patients representing both minority and nonminority populations.

[0106] The occlusal one-third of the crown was sectioned perpendicular to the long axis of the tooth using a water-cooled low-speed diamond saw (Buehler Ltd., Lake Bluff, Illinois). A uniform smear layer was created by abrading the exposed dentin surface with 600-grit silicon carbide under water. The exposed dentin surfaces were demineralized with 35% phosphoric acid for 60 seconds, rinsed with Milli-Q purified water, and sectioned perpendicular and parallel to the surface using the same water-cooled low-speed diamond saw. A diamond saw was used to make parallel cuts spaced 2 mm apart, perpendicular to the surface. A final cut was made approximately 4 mm below the flat surface. The resulting slab was approximately 8-10 mm in length, 2 mm thick, and 4 mm wide. The samples were immersed in a simulated body fluid (SBF) solution prepared following the ISO 23317:2014 standard, which had a nearly equal concentration of ions as blood plasma or body fluid (pH = 7.40) and stored at +4 °C until used. This SBF solution was used to replicate the oral environment in which the dentin or treated caries would be exposed to saliva.

[0107] SDF and AgBP Application. The University of California, San Francisco (UCSF) Caries Arrest Committee and the manufacturer’s detailed guidelines were followed to apply SDF to dental tissue. Briefly, after removing the prepared dentin slabs from the SBF and rinsing them with Milli-Q purified water, they were gently dried with compressed air briefly to expose the dental tissue and prevent dilution of the SDF solution. A single drop of commercially available 38% SDF solution was transferred to a fresh plastic dish, covered with a cap to protect from light, the SDF was gently brushed onto the dental tissue for 1 min before being rinsed with Milli-Q purified water using a micro-brush applicator. The tissue was then placed back into the modified SBF and incubated at 37 °C for 6 hours to mimic the oral environment and allow SDF activity. Darkening of the treated regions due to the precipitation of metallic silver and silver oxides on the dental tissue was observed over time.

[0108] The process of applying the AgBP to evaluate samples followed the SDF process. The peptide stocks, which were synthesized and lyophilized, were stored at -20 °C. Prior to use, the peptide stocks were reconstituted in Milli-Q purified water (Resistivity at 25 °C: 18.2 MQ cm, total organic content: < 5 ppb, filtered through a 0.22 pm filter) at a concentration of 50 pM, to ensure an adequate amount of peptide was available. The peptide solution was then drop-cast onto the dentin slabs and incubated for 2 hours at room temperature in the dark. Subsequently, the excess peptide solution was removed by gently rinsing with Milli-Q purified water, and the slabs were placed in fresh SBF solution.

[0109] Localization of AgBP on Dental Tissue. SDF -treated dentin slabs were randomly divided into three groups. The first group was stored for use as SDF-only treated dentin slabs. The second group was incubated with biotinylated AgBP, and the third group with biotinylated Spacer- AgBP, as indicated in the application protocol. Following peptide treatment, all three groups were rinsed with Milli-Q purified water, gently dried with compressed air, and 10 pL of freshly prepared 25 nM Q-dot ™ 655 Streptavidin Conjugate was drop-casted onto the treated surfaces. Samples were incubated at room temperature for 1 hour, protected from light, washed with Milli-Q purified water 3 times to remove unbound Q- dot s and images were captured by Leica TCS SPE Laser Scanning upright microscope (Model DMR-Q). A 20X Olympus UPlanSApo/0.75 with M25 x 0.75 threaded ThorLabs adapter was used for magnification on air where images were captured with the following parameters: image size of 1.1x1.1 mm, pixel size of 537.37 x 537.37 nm, pinhole size of 75.54 pm, and an optical section of 2.057 pm. Z-stack images were acquired with a 12-bit depth, resolution of 2048 x 2048, pixel dwell time of 360 ns, and a frame rate of 0.185 frames per second. The laser intensity was set at 15% with an excitation wavelength of 405 nm and an emission wavelength evaluated in the range of 643-670 nm centered at 655 nm. The collected Z-stack images were processed using the Fiji software (ImageJ, Version 1.54f) to generate maximum intensity projections and mean fluorescence intensity calculations ³⁷

[0110] Results and Discussion. FIG. 3 is a bar graph comparing mean fluorescence intensities of SDF -treated dentin, SDF and biotinylated AgBP-treated dentin, and SDF and biotinylated KGSGGG- AgBP-treated dentin after further treatment with a streptavidin- conjugated Q-dot solution. (One-way Anova, *** denotes p<0.001 (n=8, ±SEM)). This study indicated the similar binding of the AgBP peptide with or without a spacer amino acid sequence KGSGGG to SDF-treated dentinal surfaces.

[OHl] Instead of directly labeling the peptide, an indirect yet more selective approach was employed, utilizing streptavi din-conjugated Quantum Dots (Q-dot ™ 655) to bind to biotin and detect its presence in various applications. In addition to biotin-conjugated AgBP, the spacer in spacer-AgBP (KGSGGG-EQLGVRKELRGV) had a negligible effect on the binding specificity of the peptide. Dentin slabs were randomly selected and treated with SDF, followed by rinsing with Milli-Q purified water and incubating with 50 pM of either Biotin-AgBP or Biotin-Spacer-AgBP, according to their assignments. Subsequently, a streptavidin-conjugated Q-dot solution was applied, and fluorescent images were captured using a Leica confocal microscope. ImageJ (Fiji) software was used to analyze the images and calculate the mean fluorescence intensities for comparison of binding ability. The confocal microscopy images displayed in FIG. 3 show the difference in fluorescence intensity between SDF and AgBP derivatives when applied to dentin surfaces. When the Q- dots were exposed to the SDF -treated surfaces, only faint fluorescence was detected, indicating non-specific binding. However, upon the addition of either of the AgBP derivatives (biotin- AgBP or biotin-spacer- AgBP), the fluorescence intensity post-wash increased significantly, indicative of specific binding between streptavidin-conjugated Q-dot and biotin-conjugated AgBP and biotin-conjugated spacer-AgBP, respectively. These images not only indicate the localization of the silver-binding peptide, but also indicate the robust effectiveness of silver binding with a spacer sequence. The mean fluorescence intensity for SDF -treated dentin was relatively low (8.6), suggesting minimal non-specific binding. However, upon the application of biotinylated AgBP after SDF treatment, the mean fluorescence intensity increased significantly to 16.0, indicating the strong binding of the AgBP derivative to the SDF-modified dentin surface. This binding was comparable when the biotinylated spacer-AgBP construct was used, with a mean fluorescence intensity of 16.1. These findings provide evidence for the selective targeting capability of the engineered AgBP derivatives for SDF -treated tooth surfaces.

[0112] Without being bound by any theory, the application of silver diamine fluoride to tooth structures may initiate a sequence of reactions, starting with the release of silver ions from the SDF ion complex. Some of these silver ions may react with hydroxyapatite to form silver phosphate and silver oxides, whereas others may be reduced by structural proteins such as collagen to form metallic silver that subsequently forms a complex with the protein. Moreover, some silver ions may form silver halides, particularly silver chloride, by reacting with the readily available chloride ions in saliva. However, silver may be primarily replaced by silver chloride and silver oxide and a small portion may be reduced to form metallic silver. These silver compounds may be responsible for the formation of dark stains and color changes observed in tooth structures after SDF application.

[0113] Example 3: Study of Penetration of AgBP into Dentin [0114] AgBP peptide was prepared as in Example 2. Dental slabs were prepared and treated with SDF or SDF and AgBP as described in Example 2.

[0115] Micro X-ray computed tomography (Micro-XCT). The extent of silver ion penetration within the dental slabs, including those treated with SDF and SDF/AgBP, was examined using a 3D micro-X-ray computed tomography system (Micro-XCT-400, Xradia Inc., Pleasanton, CA, USA). Three samples from each group were scanned, and transmission X-ray images of the samples were captured at tungsten anode settings of 80 kV and 8 W. A total of 1000 images were obtained from a 360° rotation at a resolution of 15 seconds per image. The 3D images were reconstructed using the XM Reconstructor software (version 8.0), and the Fiji software (ImageJ, Version 1.54f) Volume Viewer plugin was used to analyze the constructed volumes. Twenty measurements were recorded from the middle section of the dental slabs.

[0116] Results and Discussion. Examination of dental slabs via visual inspection demonstrated a more pronounced penetration of silver ions. To obtain additional information, micro-X-ray computed tomography (Micro-XCT) scanning was employed on three samples from each group. This non-destructive analysis method employs three- dimensional imaging to evaluate the internal structure of dental slabs and serves as a reliable indicator of the depth of silver ion penetration. A total of 1000 images were captured, and their 3D reconstructions were accomplished using the XM Reconstructor software (Version 8.0). The resulting 3D images were then analyzed using the Volume Viewer plugin of the ImageJ software. The reconstructed 3D volume's grayscale voxels, representing X-ray attenuation, were assigned colors using a false color lookup table (LUT). This technique does not alter the underlying data but enhances visibility of specific features and subtle attenuation changes for easier human perception. Although the LUT color maps effectively illustrate differences in density through X-ray attenuation, they do not provide direct chemical composition information. Data analysis was concluded extracting cross-sections from the middle sections of the slabs and measuring the depth of each sample from 20 different points.

[0117] FIG. 4 is a bar graph of the average depth of penetration of silver ions into dental samples as measured using micro-CT images of dentin treated with SDF alone or with AgBP peptide, (t-test, n=60, ±SD, ***denotes p<0.001). This study indicated that AgBP may effectively regulate the formation of silver compounds, providing a higher concentration of and deeper penetration of silver ions into the dentin structure.

[0118] The cross-sections illustrated the successful penetration of silver ions into the dentin. A bright, dense silver layer was clearly observed, extending from the top of the slab to the dentin. The average depth of silver ion penetration was measured to be 814.8 pm (±138.01 pm) for the SDF-treated dental slabs, while the SDF/AgBP treatment resulted in a depth of 1090.7 pm (±142.3 pm). A paired t-test demonstrated a statistically significant difference (p < .001) between the two groups. AgBP may safeguard the active silver ions derived from the SDF solution against conversion into metallic silver and silver oxides, allowing them to remain active for a longer duration than the SDF-only treated surfaces. A higher concentration of active silver ions and deeper infiltration into the dentin structure was seen in the sample treated with AgBP. .

[0119] While certain embodiments have been illustrated and described, it should be understood that changes and modifications may be made therein in accordance with ordinary skill in the art without departing from the technology in its broader aspects as defined in the following claims.

[0001] The embodiments, illustratively described herein may suitably be practiced in the absence of any element or elements, limitation or limitations, not specifically disclosed herein. Thus, for example, the terms “comprising,” “including,” “containing,” etc. shall be read expansively and without limitation. Additionally, the terms and expressions employed herein have been used as terms of description and not of limitation, and there is no intention in the use of such terms and expressions of excluding any equivalents of the features shown and described or portions thereof, but it is recognized that various modifications are possible within the scope of the claimed technology. Additionally, the phrase “consisting essentially of’ will be understood to include those elements specifically recited and those additional elements that do not materially affect the basic and novel characteristics of the claimed technology. The phrase “consisting of’ excludes any element not specified.

[0002] The present disclosure is not to be limited in terms of the particular embodiments described in this application. Many modifications and variations may be made without departing from its spirit and scope, as will be apparent to those skilled in the art. Functionally equivalent methods and compositions within the scope of the disclosure, in addition to those enumerated herein, will be apparent to those skilled in the art from the foregoing descriptions. Such modifications and variations are intended to fall within the scope of the appended claims. The present disclosure is to be limited only by the terms of the appended claims, along with the full scope of equivalents to which such claims are entitled. It is to be understood that this disclosure is not limited to particular methods, reagents, compounds, or compositions, which may of course vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting.

[0003] In addition, where features or aspects of the disclosure are described in terms of Markush groups, those skilled in the art will recognize that the disclosure is also thereby described in terms of any individual member or subgroup of members of the Markush group.

[0004] As will be understood by one skilled in the art, for any and all purposes, particularly in terms of providing a written description, all ranges disclosed herein also encompass any and all possible subranges and combinations of subranges thereof. Any listed range can be easily recognized as sufficiently describing and enabling the same range being broken down into at least equal halves, thirds, quarters, fifths, tenths, etc. As a non-limiting example, each range discussed herein can be readily broken down into a lower third, middle third and upper third, etc. As will also be understood by one skilled in the art all language such as “up to,” “at least,” “greater than,” “less than,” and the like, include the number recited and refer to ranges which can be subsequently broken down into subranges as discussed above. Finally, as will be understood by one skilled in the art, a range includes each individual member.

[0005] All publications, patent applications, issued patents, and other documents referred to in this specification are herein incorporated by reference as if each individual publication, patent application, issued patent, or other document was specifically and individually indicated to be incorporated by reference in its entirety. Definitions that are contained in text incorporated by reference are excluded to the extent that they contradict definitions in this disclosure.

[0006] The present technology may include, but is not limited to, the features and combinations of features recited in the following lettered paragraphs, it being understood that the following paragraphs should not be interpreted as limiting the scope of the claims as appended hereto or mandating that all such features must necessarily be included in such claims: A. A functionalized peptide of Formula I:

or a pharmaceutically acceptable salt thereof and/or a solvate thereof, wherein R¹ is hydrogen, unsubstituted Ci-Ce alkyl, or cyano; and

Y¹ is a terminal nitrogen, side chain nitrogen, or side chain sulfur of amino acid sequence

X1-X2-EQLGVRKELRGV (SEQ ID NO: 1) where Xi is absent or amino acid K, S, R, or E; and X2 is absent or a spacer of 1, 2, 3, 4, 5 ,6 , 7, 8, 9, or 10 amino acid residues.

B. The functionalized peptide of paragraph A, wherein X2 is selected from EAAAK (SEQ ID

NO: 2), APA (SEQ ID NO: 3), GGG (SEQ ID NO: 4), PAPAP (SEQ ID NO: 5), or GSGGG (SEQ ID NO: 6).

C. The functionalized peptide of paragraph A or B, wherein the amino acid sequence is

EAAAKEQLGVRKELRGV (SEQ ID NO: 9).

D The functionalized peptide of paragraph A or B, wherein the amino acid sequence is EQLGVRKELRGV (SEQ ID NO: 10).

E. The functionalized peptide of paragraph A or B, wherein the amino acid sequence is

APAKEQLGVRKELRGV (SEQ ID NO: 11).

F. The functionalized peptide of paragraph A or B, wherein the amino acid sequence is

GGGKEQLGVRKELRGV (SEQ ID NO: 12).

G. The functionalized peptide of paragraph A or B, wherein the amino acid sequence is

PAPAPKEQLGVRKELRGV (SEQ ID NO: 13).

H. The functionalized peptide of paragraph A or B, wherein the amino acid sequence is

GS GGGKEQLGVRKELRGV (SEQ ID NO: 14). I. A composition comprising the functionalized peptide of any one of paragraphs A-H; a dental adhesive; a photoinitiator; and a pharmaceutically acceptable carrier.

J. The composition of paragraph I, wherein the composition comprises an effective amount of the functionalized peptide for treating a dental surface.

K. The composition of paragraph I or J, comprising about 1 wt.% to about 50 wt.% of the functionalized peptide; about 1 wt.% to about 90 wt.% of the dental adhesive; and about 0.1 wt.% to about 10 wt.% photoinitiator.

L. The composition of any one of paragraphs I to K, wherein the dental adhesive comprises

2,2-bis[4-(2-hydroxyl-3-methacryloxypropoxy)phenyl] propane; 2-hydroxyethyl methacrylate (HEMA); propane-2, 2-diylbis [4,1 -phenyleneoxy (2 -hy droxypropane-3,1 - diyl)] bis(2-methylprop-2-enoate), 2-(methacryloyloxy)ethyl (2- (trimethylammonio)ethyl) phosphate (MPC); urethane dimethacrylate (UDMA); 3- trimethoxysilyl propyl methacrylate (MPS); (trimethoxysilyl)methyl methacrylate (MMeS); 4,4-diethoxy-9-oxo-3,10-dioxa-8-aza-4-siladodecan-12-yl methacrylate; trimethoxy silylpropyl methacrylate; a compound having the structure of Formula X

acrylic acid; methacrylic acid, ethyl acrylate; methyl acrylate; butyl acrylate; 2-ethylhexyl acrylate; hydroxy ethyl acrylate; hydroxypropyl acrylate; N-butyl acrylate; 1,3 -butanediol dimethacrylate; 1,6-hexanediol dimethacrylate, or a combination of any two or more thereof.

M. The composition of any one of paragraphs I to L, wherein the photoinitiator comprises camphoroquinone, ethyl-4-(dimethylamine) benzoate, diphenyliodonoium hexafluorophyosphate, or a combination of two or more thereof.

N. A method of treating a dental surface, the method comprising: administering silver diamine fluoride (SDF) to the dental surface; and after administering the SDF, administering an effective amount of a functionalized peptide of any one of paragraphs A to H to the dental surface.

O. The method of paragraph N, wherein the dental surface is a dental enamel and/or dentin.

P. The method of paragraph O, wherein the dental enamel and/or dentin comprises a carious region, a hypomineralized region, or both a carious region and a hypomineralized region.

Q. The method of paragraph N, wherein administering SDF to the dental surface comprises administering a solution comprising SDF to the dental surface.

R. The method of paragraph Q, wherein the solution has an SDF concentration of about 38% w/v.

S. The method of paragraph N, wherein administering the effective amount of the functionalized peptide comprises contacting the dental surface with the functionalized peptide for about 1 minute to about 4 hours.

T. The method of paragraph N, further comprising, after administering the functionalized peptide, exposing the dental surface to light to initiate polymerization of the functionalized peptide and the dental adhesive.

U. The method of paragraph N, wherein administering the effective amount of the functionalized peptide reduces visible discoloration on the dental surface resulting from administering the SDF.

V. A method of treating a dental surface, the method comprising: administering silver diamine fluoride (SDF) to the dental surface; and after administering the SDF, administering an effective amount of the composition of any one of paragraphs I to M to the dental surface.

W. The method of paragraph V, wherein the dental surface is a dental enamel and/or dentin. X. The method of paragraph W, wherein the dental enamel and/or dentin comprises a carious region, a hypomineralized region, or both a carious region and a hypomineralized region.

Y. The method of paragraph V, wherein administering SDF to the dental surface comprises administering a solution comprising SDF to the dental surface.

Z. The method of paragraph Y, wherein the solution has an SDF concentration of about 38% w/v.

AA. The method of paragraph V, wherein administering the effective amount of the composition comprises contacting the dental surface with the compound for about 1 minute to about 4 hours.

AB. The method of paragraph V, further comprising, after administering the functionalized peptide, exposing the dental surface to light to initiate polymerization of the functionalized peptide and the dental adhesive.

AC. The method of paragraph AB, wherein the light has a wavelength of about 200 nm to about 500 nm.

AD. The method of paragraph AB, wherein administering the effective amount of the composition reduces visible discoloration on the dental surface resulting from administering the SDF.

[0120] Other embodiments are set forth in the following claims, along with the full scope of equivalents to which such claims are entitled.

Claims

WHAT IS CLAIMED IS:

1. A functionalized peptide of Formula I:

or a pharmaceutically acceptable salt thereof and/or a solvate thereof, wherein R¹ is hydrogen, unsubstituted Ci-Ce alkyl, or cyano; and

Y¹ is a terminal nitrogen, side chain nitrogen, or side chain sulfur of amino acid sequence

X1-X2-EQLGVRKELRGV (SEQ ID NO: 1) where Xi is absent or amino acid K, S, R, or E; and X2 is absent or a spacer of 1, 2, 3, 4, 5 ,6 , 7, 8, 9, or 10 amino acid residues.

2. The functionalized peptide of claim 1, wherein X2 is selected from EAAAK (SEQ ID NO:

2), APA (SEQ ID NO: 3), GGG (SEQ ID NO: 4), PAPAP (SEQ ID NO: 5), or GSGGG (SEQ ID NO: 6).

3. The functionalized peptide of claim 1, wherein the amino acid sequence is

EAAAKEQLGVRKELRGV (SEQ ID NO: 9).

4. The functionalized peptide of claim 1, wherein the amino acid sequence is

EQLGVRKELRGV (SEQ ID NO: 10).

5. The functionalized peptide of claim 1, wherein the amino acid sequence is

APAKEQLGVRKELRGV (SEQ ID NO: 11).

6. The functionalized peptide of claim 1, wherein the amino acid sequence is

GGGKEQLGVRKELRGV (SEQ ID NO: 12).

7. The functionalized peptide of claim 1, wherein the amino acid sequence is

PAPAPKEQLGVRKELRGV (SEQ ID NO: 13).

8. The functionalized peptide of claim 1, wherein the amino acid sequence is

GSGGGKEQLGVRKELRGV (SEQ ID NO: 14).

9. A composition comprising the functionalized peptide of any one of claims 1-8; a dental adhesive; a photoinitiator; and a pharmaceutically acceptable carrier.

10. The composition of claim 9, wherein the composition comprises an effective amount of the functionalized peptide for treating a dental surface.

11. The composition of claim 9, comprising about 1 wt.% to about 50 wt.% of the functionalized peptide; about 1 wt.% to about 90 wt.% of the dental adhesive; and about 0.1 wt.% to about 10 wt.% photoinitiator.

12. The composition of any one of claims 9-11, wherein the dental adhesive comprises 2,2- bis[4-(2-hydroxyl-3-methacryloxypropoxy)phenyl] propane; 2-hydroxyethyl methacrylate (HEMA); propane-2, 2-diylbis [4,1 -phenyleneoxy (2 -hy droxypropane-3,1 - diyl)] bis(2-methylprop-2-enoate), 2-(methacryloyloxy)ethyl (2- (trimethylammonio)ethyl) phosphate (MPC); urethane dimethacrylate (UDMA); 3- trimethoxysilyl propyl methacrylate (MPS); (trimethoxysilyl)methyl methacrylate (MMeS); 4,4-diethoxy-9-oxo-3,10-dioxa-8-aza-4-siladodecan-12-yl methacrylate; trimethoxy silylpropyl methacrylate; a compound having the structure of Formula X

acrylic acid; methacrylic acid, ethyl acrylate; methyl acrylate; butyl acrylate; 2-ethylhexyl acrylate; hydroxy ethyl acrylate; hydroxypropyl acrylate; N-butyl acrylate; 1,3 -butanediol dimethacrylate; 1,6-hexanediol dimethacrylate, or a combination of any two or more thereof.

13. The composition of any one of claims 9-12, wherein the photoinitiator comprises camphoroquinone, ethyl-4-(dimethylamine) benzoate, diphenyliodonoium hexafluorophyosphate, or a combination of two or more thereof.

14. A method of treating a dental surface, the method comprising: administering silver diamine fluoride (SDF) to the dental surface; and after administering the SDF, administering an effective amount of a functionalized peptide of any one of claims 1-8 to the dental surface.

15. The method of claim 14, wherein the dental surface is a dental enamel and/or dentin.

16. The method of claim 15, wherein the dental enamel and/or dentin comprises a carious region, a hypomineralized region, or both a carious region and a hypomineralized region.

17. The method of claim 14, wherein administering SDF to the dental surface comprises administering a solution comprising SDF to the dental surface.

18. The method of claim 17, wherein the solution has an SDF concentration of about 38% w/v.

19. The method of claim 14, wherein administering the effective amount of the functionalized peptide comprises contacting the dental surface with the functionalized peptide for about 1 minute to about 4 hours.

20. The method of claim 14, further comprising, after administering the functionalized peptide, exposing the dental surface to light to initiate polymerization of the functionalized peptide and the dental adhesive.

21. The method of claim 14, wherein administering the effective amount of the functionalized peptide reduces visible discoloration on the dental surface resulting from administering the SDF.

22. A method of treating a dental surface, the method comprising: administering silver diamine fluoride (SDF) to the dental surface; and after administering the SDF, administering an effective amount of the composition of any one of claims 9-13 to the dental surface.

23. The method of claim 22, wherein the dental surface is a dental enamel and/or dentin.

24. The method of claim 23, wherein the dental enamel and/or dentin comprises a carious region, a hypomineralized region, or both a carious region and a hypomineralized region.

25. The method of claim 22, wherein administering SDF to the dental surface comprises administering a solution comprising SDF to the dental surface.

26. The method of claim 25, wherein the solution has an SDF concentration of about 38% w/v.

27. The method of claim 22, wherein administering the effective amount of the composition comprises contacting the dental surface with the compound for about 1 minute to about 4 hours.

28. The method of claim 22, further comprising, after administering the functionalized peptide, exposing the dental surface to light to initiate polymerization of the functionalized peptide and the dental adhesive.

29. The method of claim 28, wherein the light has a wavelength of about 200 nm to about

500 nm.

30. The method of claim 22, wherein administering the effective amount of the composition reduces visible discoloration on the dental surface resulting from administering the SDF.