# US 10,441,667 B2 · Composition and method for detection of demineralisation > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 10,441,667 B2 - Document ID: US10441667B2 - Status: Granted - Kind: B2 - Application number: 12/520,128 - Filed: 2007-12-21 - Published: 2010-02-11 - Granted: 2019-10-15 - Earliest claimed: 2006-12-21 - Assignees: Calcivis Limited - Applicants: Emma Perfect; Chris Longbottom - Inventors: Emma Perfect; Chris Longbottom - Web page: https://patentgrep.com/patent/US10441667B2 ## CPC - A61K 49/006: {Biological staining of tissues in vivo, e.g. methylene blue or toluidine blue O administered in the buccal area to detect epithelial cancer cells, dyes used for delineating tissues during surgery} - A61P 1/02: Stomatological preparations, e.g. drugs for caries, aphtae, periodontitis - A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D) - A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS ## Abstract The invention relates to a composition for the detection of tooth demineralization. More specifically, the invention concerns a composition comprising a complex capable of producing an optical signal characteristic of the presence of free ions, the pharmaceutical uses of such a composition, and methods and a kit for the detection of active demineralization at tooth surfaces using such a composition. ## Claims ### Claim 1 An intra-oral pharmaceutical tooth demineralization detection composition, comprising: a) 1 ng/mL to 10 mg/mL of a pharmaceutically acceptable concentration of proteins and protein complexes which produce a bioluminescent optical signal in the presence of free ions and b) a pharmaceutically acceptable concentration of an ion chelator that gives a significantly high signal to noise ratio, wherein the tooth demineralization detection composition is applied to at least one tooth in the mouth of a living patient. ### Claim 2 (depends on 1) A composition according to claim 1, wherein at least one ion to be detected is selected from the group consisting of calcium ions, magnesium ions, phosphate ions, carbonate ions, potassium ions, strontium ions, fluoride ions, copper ions, chloride ions, zinc ions, lead ions, tin ions, iron ions or enamelins. ### Claim 3 (depends on 1) A composition according to claim 1, further comprising a pharmaceutically acceptable additive. ### Claim 4 (depends on 3) A composition according to claim 3, wherein the additive is a therapeutically active agent. ### Claim 5 (depends on 4) A composition according to claim 4, wherein the active agent is a bactericidal or bacteriostatic agent. ### Claim 6 (depends on 1) A composition according to claim 1, further comprising a pharmaceutically acceptable excipient. ### Claim 7 (depends on 6) A composition according to claim 6, wherein the excipient is a flavouring or a colouring additive. ### Claim 8 (depends on 1) A composition according to claim 1, wherein the composition is in a pharmaceutically acceptable dosage form. ### Claim 9 (depends on 8) A composition according to claim 8, wherein the composition is a liquid, a powder, a mouldable substance or a gel. ### Claim 10 (depends on 1) A composition according to claim 1, wherein the protein or protein complex undergoes a conformation change on binding free ions that leads to an optical signal being generated. ### Claim 11 (depends on 1) A composition according to claim 1, wherein the protein or protein complex comprises aequorin, obelin, clytin, mitrocomin, halistaurin, phialidin, mnemiopsin, symplectin, gr-bolinopsin, casein, calsequestrin, calexcitin, calcium binding cysteine protease, calmodulin, berovin, or mixtures thereof. ### Claim 12 (depends on 1) A composition according to claim 1, wherein the protein or protein complex comprises tandem fusions of a fluorescent protein, calmodulin and a calmodulin-binding peptide. ### Claim 13 (depends on 1) A composition according to claim 1, wherein the complex fluoresces at a characteristic wavelength indicative of the presence of ions. ### Claim 14 (depends on 1) A composition according to claim 1, wherein the composition comprises a first complex which produces an optical signal on binding with free ions, and a second complex which fluoresces at a characteristic wavelength indicative of the presence of free ions. ### Claim 15 (depends on 1) A composition according to claim 1, wherein the protein or protein complex is modified to enhance the signal or alter its emission spectra. ### Claim 16 (depends on 15) A composition according to claim 15, wherein the modification of the protein or protein complex is accomplished through alteration of the DNA sequence of the gene, by acetylation, ethoxycarbonylation, fluorescamine-modification, fluorescein labelling or by creation of chimeric proteins. ### Claim 17 (depends on 1) A composition according to claim 1, wherein the protein or protein complex comprises a recombinant protein. ### Claim 18 (depends on 1) A composition according to claim 1, wherein the composition is optically transparent. ### Claim 19 (depends on 1) A composition according to claim 1, wherein the optical signal produced by the composition is detectable using a spectrophotometer, charge coupled device (CCD), complementary metal-oxide semiconductor CMOS, digital camera, intensified camera, intraoral camera, videoscope, photographic film, fibre-optic device, photometric detector, photomultiplier, micro-electro-mechanical system (MEMS) or visually by eye. ### Claim 20 (depends on 1) A composition according to claim 1, wherein the ion chelator is a calcium chelator. ### Claim 21 (depends on 20) A composition according to claim 20, wherein the calcium chelator is EDTA. ### Claim 22 An intra-oral pharmaceutical tooth demineralization detection composition, comprising proteins and protein complexes which produce a bioluminescent optical signal in the presence of free ions, wherein the bioluminescent optical signal is detected intra-orally, wherein the composition comprises about 1 ng to about 10 mg/mL of protein or protein complex, and wherein the protein or protein complex is modified to enhance the signal or alter its emission spectra through alteration of the DNA sequence of the gene coding for the protein or protein complex wherein the tooth demineralization detection composition is applied to at least one tooth in the mouth of a living patient. ### Claim 23 (depends on 22) A composition according to claim 22, further comprising a pharmaceutically acceptable additive. ### Claim 24 (depends on 23) A composition according to claim 23, wherein the additive is a therapeutically active agent. ### Claim 25 (depends on 24) A composition according to claim 24, wherein the active agent is a bactericidal or bacteriostatic agent. ### Claim 26 (depends on 22) A composition according to claim 22, further comprising a pharmaceutically acceptable excipient. ### Claim 27 (depends on 26) A composition according to claim 26, wherein the excipient is a flavouring or a colouring additive. ### Claim 28 (depends on 22) A composition according to claim 22, wherein the composition is in a pharmaceutically acceptable dosage form. ### Claim 29 (depends on 28) A composition according to claim 28, wherein the composition is a gel. ### Claim 30 (depends on 22) A composition according to claim 22, wherein the protein or protein complex undergoes a conformation change on binding free ions that leads to an optical signal being generated. ### Claim 31 (depends on 22) A composition according to claim 22, wherein the protein or protein complex comprises aequorin, obelin, clytin, mitrocomin, halistaurin, phialidin, mnemiopsin, berovin, or mixtures thereof. ### Claim 32 (depends on 22) A composition according to claim 22, wherein the protein or protein complex comprises a recombinant protein. ### Claim 33 (depends on 22) A composition according to claim 22, wherein the optical signal produced by the composition is detectable using a spectrophotometer, charge coupled device (CCD), complementary metal-oxide semiconductor CMOS, digital camera, intensified camera, intraoral camera, videoscope, photographic film, fibre-optic device, photometric detector, photomultiplier, micro-electro-mechanical system (MEMS) or visually by eye. ### Claim 34 (depends on 22) A composition according to claim 22, wherein the DNA sequence of the gene coding for the protein or protein complex is altered by acetylation, ethoxycarbonylation, fluorescamine-modification, fluorescein labeling, or by creation of chimeric proteins. ### Claim 35 An intra-oral pharmaceutical tooth demineralization detection composition, comprising a complex capable of producing an optical signal characteristic of the presence of free ions, wherein the complex comprises: a) a pharmaceutically acceptable concentration of proteins and protein complexes which produce a bioluminescent optical signal in the presence of free ions wherein the protein or protein complex comprises aequorin, obelin, clytin, mitrocomin, halistaurin, phialidin, mnemiopsin, symplectin, gr-bolinopsin, casein, calsequestrin, calexcitin, calcium binding cysteine protease, calmodulin, berovin, or mixtures thereof; and b) up to 1 mM EDTA, wherein the composition comprises from 1 ng/ml to 10 mg/ml of complex and gives a significantly high signal to noise ratio.