CN116440077A - 一种预防及快速诊断和治疗龋病的口腔喷剂及其制备方法 - Google Patents
一种预防及快速诊断和治疗龋病的口腔喷剂及其制备方法 Download PDFInfo
- Publication number
- CN116440077A CN116440077A CN202310267812.1A CN202310267812A CN116440077A CN 116440077 A CN116440077 A CN 116440077A CN 202310267812 A CN202310267812 A CN 202310267812A CN 116440077 A CN116440077 A CN 116440077A
- Authority
- CN
- China
- Prior art keywords
- meotpy
- caries
- oral spray
- washing
- preventing
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 208000002925 dental caries Diseases 0.000 title claims abstract description 38
- 229940041678 oral spray Drugs 0.000 title claims abstract description 35
- 239000000668 oral spray Substances 0.000 title claims abstract description 35
- 238000002360 preparation method Methods 0.000 title claims abstract description 8
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims abstract description 28
- 238000000034 method Methods 0.000 claims abstract description 22
- TVXBFESIOXBWNM-UHFFFAOYSA-N Xylitol Natural products OCCC(O)C(O)C(O)CCO TVXBFESIOXBWNM-UHFFFAOYSA-N 0.000 claims abstract description 8
- HEBKCHPVOIAQTA-UHFFFAOYSA-N meso ribitol Natural products OCC(O)C(O)C(O)CO HEBKCHPVOIAQTA-UHFFFAOYSA-N 0.000 claims abstract description 8
- 239000003068 molecular probe Substances 0.000 claims abstract description 8
- 239000000811 xylitol Substances 0.000 claims abstract description 8
- 235000010447 xylitol Nutrition 0.000 claims abstract description 8
- HEBKCHPVOIAQTA-SCDXWVJYSA-N xylitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)CO HEBKCHPVOIAQTA-SCDXWVJYSA-N 0.000 claims abstract description 8
- 229960002675 xylitol Drugs 0.000 claims abstract description 8
- 239000007853 buffer solution Substances 0.000 claims abstract description 7
- 230000008569 process Effects 0.000 claims abstract description 5
- 125000004172 4-methoxyphenyl group Chemical group [H]C1=C([H])C(OC([H])([H])[H])=C([H])C([H])=C1* 0.000 claims abstract description 4
- 229920002554 vinyl polymer Polymers 0.000 claims abstract description 4
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 20
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 14
- 238000005406 washing Methods 0.000 claims description 11
- 239000000543 intermediate Substances 0.000 claims description 10
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 9
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 9
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 9
- 238000006243 chemical reaction Methods 0.000 claims description 7
- 238000001035 drying Methods 0.000 claims description 7
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 6
- NQRYJNQNLNOLGT-UHFFFAOYSA-N Piperidine Chemical compound C1CCNCC1 NQRYJNQNLNOLGT-UHFFFAOYSA-N 0.000 claims description 6
- 239000007787 solid Substances 0.000 claims description 6
- 239000000243 solution Substances 0.000 claims description 6
- CTRXZOLNEVJBDX-UHFFFAOYSA-N 4-(4-methoxy-n-(4-methoxyphenyl)anilino)benzaldehyde Chemical compound C1=CC(OC)=CC=C1N(C=1C=CC(C=O)=CC=1)C1=CC=C(OC)C=C1 CTRXZOLNEVJBDX-UHFFFAOYSA-N 0.000 claims description 5
- 239000000872 buffer Substances 0.000 claims description 5
- 239000002904 solvent Substances 0.000 claims description 5
- 238000001914 filtration Methods 0.000 claims description 4
- 239000000376 reactant Substances 0.000 claims description 4
- 238000003756 stirring Methods 0.000 claims description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 4
- DXTDZBSTWOXMNU-UHFFFAOYSA-N 4-iodo-2,6-dimethylpyridine Chemical compound CC1=CC(I)=CC(C)=N1 DXTDZBSTWOXMNU-UHFFFAOYSA-N 0.000 claims description 3
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 3
- 229910052757 nitrogen Inorganic materials 0.000 claims description 3
- 238000010992 reflux Methods 0.000 claims description 3
- 229920006395 saturated elastomer Polymers 0.000 claims description 3
- 239000000741 silica gel Substances 0.000 claims description 3
- 229910002027 silica gel Inorganic materials 0.000 claims description 3
- 238000004809 thin layer chromatography Methods 0.000 claims description 3
- 238000001291 vacuum drying Methods 0.000 claims description 3
- ZFAGSDJDJKIGEV-UHFFFAOYSA-N 1,4-dimethyl-2H-pyridine hydroiodide Chemical compound I.CN1CC=C(C)C=C1 ZFAGSDJDJKIGEV-UHFFFAOYSA-N 0.000 claims description 2
- 239000003054 catalyst Substances 0.000 claims description 2
- 238000001816 cooling Methods 0.000 claims description 2
- 238000000746 purification Methods 0.000 claims description 2
- 238000000967 suction filtration Methods 0.000 claims description 2
- 238000001308 synthesis method Methods 0.000 claims description 2
- 239000008055 phosphate buffer solution Substances 0.000 claims 1
- 241000894006 Bacteria Species 0.000 abstract description 19
- 238000011282 treatment Methods 0.000 abstract description 15
- 238000003745 diagnosis Methods 0.000 abstract description 12
- 210000000214 mouth Anatomy 0.000 abstract description 3
- 241000194019 Streptococcus mutans Species 0.000 description 18
- 230000001580 bacterial effect Effects 0.000 description 12
- 244000052616 bacterial pathogen Species 0.000 description 11
- 241000588724 Escherichia coli Species 0.000 description 10
- XDFNWJDGWJVGGN-UHFFFAOYSA-N 2-(2,7-dichloro-3,6-dihydroxy-9h-xanthen-9-yl)benzoic acid Chemical compound OC(=O)C1=CC=CC=C1C1C2=CC(Cl)=C(O)C=C2OC2=CC(O)=C(Cl)C=C21 XDFNWJDGWJVGGN-UHFFFAOYSA-N 0.000 description 8
- 238000000799 fluorescence microscopy Methods 0.000 description 7
- 229920001817 Agar Polymers 0.000 description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- 239000008272 agar Substances 0.000 description 4
- 230000000844 anti-bacterial effect Effects 0.000 description 4
- 238000005286 illumination Methods 0.000 description 4
- 239000003642 reactive oxygen metabolite Substances 0.000 description 4
- 239000000523 sample Substances 0.000 description 4
- 208000025157 Oral disease Diseases 0.000 description 3
- 238000002189 fluorescence spectrum Methods 0.000 description 3
- 208000030194 mouth disease Diseases 0.000 description 3
- 230000004044 response Effects 0.000 description 3
- 230000001954 sterilising effect Effects 0.000 description 3
- 238000004659 sterilization and disinfection Methods 0.000 description 3
- 229910019142 PO4 Inorganic materials 0.000 description 2
- 239000003242 anti bacterial agent Substances 0.000 description 2
- 229940088710 antibiotic agent Drugs 0.000 description 2
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 238000001218 confocal laser scanning microscopy Methods 0.000 description 2
- 238000003384 imaging method Methods 0.000 description 2
- 208000015181 infectious disease Diseases 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 239000001301 oxygen Substances 0.000 description 2
- 229910052760 oxygen Inorganic materials 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 2
- 239000010452 phosphate Substances 0.000 description 2
- 230000002265 prevention Effects 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 239000011550 stock solution Substances 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 208000002064 Dental Plaque Diseases 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- 206010018367 Glomerulonephritis chronic Diseases 0.000 description 1
- 201000004328 Pulpitis Diseases 0.000 description 1
- 206010037464 Pulpitis dental Diseases 0.000 description 1
- 206010040047 Sepsis Diseases 0.000 description 1
- 241000194017 Streptococcus Species 0.000 description 1
- 208000008312 Tooth Loss Diseases 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 208000022362 bacterial infectious disease Diseases 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 238000007374 clinical diagnostic method Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 210000003464 cuspid Anatomy 0.000 description 1
- 210000003298 dental enamel Anatomy 0.000 description 1
- 238000002405 diagnostic procedure Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000013399 early diagnosis Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 206010014665 endocarditis Diseases 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 238000001506 fluorescence spectroscopy Methods 0.000 description 1
- 235000003599 food sweetener Nutrition 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000002458 infectious effect Effects 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000004949 mass spectrometry Methods 0.000 description 1
- 238000001819 mass spectrum Methods 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 208000004480 periapical periodontitis Diseases 0.000 description 1
- 239000000825 pharmaceutical preparation Substances 0.000 description 1
- 230000004962 physiological condition Effects 0.000 description 1
- 239000002798 polar solvent Substances 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 238000011321 prophylaxis Methods 0.000 description 1
- 238000002390 rotary evaporation Methods 0.000 description 1
- 239000012047 saturated solution Substances 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 239000007921 spray Substances 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 230000001502 supplementing effect Effects 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 239000003765 sweetening agent Substances 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 230000036962 time dependent Effects 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 238000002604 ultrasonography Methods 0.000 description 1
- 238000011179 visual inspection Methods 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0053—Mouth and digestive tract, i.e. intraoral and peroral administration
- A61K9/006—Oral mucosa, e.g. mucoadhesive forms, sublingual droplets; Buccal patches or films; Buccal sprays
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
- A61K41/0057—Photodynamic therapy with a photosensitizer, i.e. agent able to produce reactive oxygen species upon exposure to light or radiation, e.g. UV or visible light; photocleavage of nucleic acids with an agent
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/20—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing sulfur, e.g. dimethyl sulfoxide [DMSO], docusate, sodium lauryl sulfate or aminosulfonic acids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/26—Carbohydrates, e.g. sugar alcohols, amino sugars, nucleic acids, mono-, di- or oligo-saccharides; Derivatives thereof, e.g. polysorbates, sorbitan fatty acid esters or glycyrrhizin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/001—Preparation for luminescence or biological staining
- A61K49/0013—Luminescence
- A61K49/0017—Fluorescence in vivo
- A61K49/0019—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules
- A61K49/0021—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules the fluorescent group being a small organic molecule
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/001—Preparation for luminescence or biological staining
- A61K49/0013—Luminescence
- A61K49/0017—Fluorescence in vivo
- A61K49/005—Fluorescence in vivo characterised by the carrier molecule carrying the fluorescent agent
- A61K49/0052—Small organic molecules
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/10—Dispersions; Emulsions
- A61K9/12—Aerosols; Foams
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/02—Stomatological preparations, e.g. drugs for caries, aphtae, periodontitis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/02—Local antiseptics
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D213/00—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members
- C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
- C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
- C07D213/24—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with substituted hydrocarbon radicals attached to ring carbon atoms
- C07D213/36—Radicals substituted by singly-bound nitrogen atoms
- C07D213/38—Radicals substituted by singly-bound nitrogen atoms having only hydrogen or hydrocarbon radicals attached to the substituent nitrogen atom
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Dispersion Chemistry (AREA)
- Molecular Biology (AREA)
- Biochemistry (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Biomedical Technology (AREA)
- Physiology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Nutrition Science (AREA)
- Communicable Diseases (AREA)
- Oncology (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
本发明公开了一种防治龋病的口腔喷剂的制备方法，将分子探针[4‑(2‑{4‑[双‑(4‑甲氧基‑苯基)‑氨基]‑苯基}‑乙烯基)‑1‑甲基‑吡啶鎓]+PF6 ‑(MeOTpy)和木糖醇溶于含有体积分数为0.1％的二甲基亚砜(DMSO)的PBS缓冲溶液中，即得防治龋病的口腔喷剂；所述分子探针的结构式为：本发明提供的口腔喷剂能够快速有效的实时成像和杀死口腔细菌生物膜，防治龋病，清新口气。同时又避免了杀死口腔中的一些共生菌，降低龋病诊疗费用，简化了诊疗过程，在龋病诊断、治疗中显示出良好的前景。
Description
技术领域
本发明涉及药物制剂技术领域，具体涉及一种预防及快速诊断和治疗龋病的口腔喷剂及其制备方法。
背景技术
公开该背景技术部分的信息仅仅旨在增加对本发明的总体背景的理解，而不必然被视为承认或以任何形式暗示该信息构成已经成为本领域一般技术人员所公知的现有技术。
龋病是一种常见的细菌感染性口腔疾病，口腔细菌会粘附在牙齿表面上形成的细菌生物膜，其主要的致病菌是变形链球菌(S.mutans)。严重的龋坏可能会引发其他口腔疾病，如牙髓炎、根尖周炎和牙齿脱落等。此外，口腔疾病可导致局部感染的传播，如慢性肾炎、心内膜炎和脓毒症，因此龋病的早期诊疗对人类健康至关重要。
随着科学技术的发展，不断有抗生素被开发应用于细菌感染性疾病的治疗，然而，随着抗生素的过度滥用，促使细菌进化从而产生耐药性。再者，目前临床诊断方法局限于牙科医生使用锋利的尖牙探针进行详细的目视检查，缺乏客观性，不可避免地会损伤组织。其他支持诊断方法如影像学检查、染色技术和超声技术，这些方法昂贵、复杂，且耗时长。因此发展一种简便、成本低、快速有效的诊断和治疗龋病的手段是有必要的。
发明内容
为了克服上述问题，本发明提供了一种预防及快速诊断和治疗龋病的口腔喷剂及其制备方法。
为实现上述技术目的，本发明采用如下技术方案：
本发明的第一个方面，提供了一种预防及快速诊断和治疗龋病的口腔喷剂的制备方法，将分子探针[4-(2-{4-[双-(4-甲氧基-苯基)-氨基]-苯基}-乙烯基)-1-甲基-吡啶鎓]+PF6 -(MeOTpy)和木糖醇溶于含有二甲基亚砜(DMSO)的磷酸盐(PBS)缓冲溶液中，即得预防及快速诊断和治疗龋病的口腔喷剂；
所述分子探针的结构式为：
本发明的第二个方面，提供由上述口腔喷剂的制备方法制备的口腔喷剂。
本发明的有益效果在于：
(1)本发明制备的探针分子，合成方法简单，能够快速与龋病致病菌S.mutans结合，在白光照射下结合细菌的分子发射荧光并原位产生活性氧，同时实现致病菌的成像与杀灭。
(2)木糖醇是一种甜味剂，具有清新口气、补充能量、促进唾液分泌的功能，同时木糖醇不能被口腔链球菌细菌发酵利用，可以抑制细菌生长和酸的产生从而预防龋齿，将木糖醇和探针分子联合应用于制备口腔喷剂，实现了龋病诊断、预防和治疗的临床目标。
二甲基亚砜DMSO是一种医用级别极性溶剂，提高探针分子溶解度，于水混溶利于喷剂的稳定性，同时具有一定的消炎和局部止痛的作用
(3)本发明提供的口腔喷剂能够对龋病致病菌进行快速有效的实时成像和杀死口腔龋病致病菌和控制龋病生物膜，防治龋病，清新口气。同时又避免了杀死口腔中的一些共生菌，实现精准治疗。同时，降低龋病诊疗费用，简化了诊疗过程，在龋病诊断、治疗中显示出良好的前景。
附图说明
构成本申请的一部分的说明书附图用来提供对本申请的进一步理解，本申请的示意性实施例及其说明用于解释本申请，并不构成对本申请的不当限定。
图1为MeOTpy质谱图；
图2为MeOTpy对致病菌S.mutans和一种共生菌大肠杆菌(E.coli)在30S、5min、10min和20min快速荧光成像图；
图3为MeOTpy对致病菌S.mutans的荧光响应动力学实验图；
图4为光动力产生活性氧的图像，其中A为MeOTpy在光照条件下随时间的变化的荧光光谱图；B为DCFH在光照条件下随时间的变化的荧光光谱图；C为MeOTpy和DCFH在光照条件下随时间的变化的荧光光谱图；D为MeOTpy、DCFH及MeOTpy+DCFH在光照条件下随时间的变化的荧光强度数据输出图像；
图5为MeOTpy光动力杀死致病菌图像，其中A为MeOTpy对致病菌变形链球菌的杀菌图；B为MeOTpy对致病菌变形链球菌的杀菌率数据输出图；C为MeOTpy对共生菌大肠杆菌的杀菌图；D为MeOTpy对共生菌大肠杆菌的杀菌率数据输出图；
图6为本发明口腔喷剂荧光成像图，包括健康和龋坏的离体人牙，A为本发明口腔喷剂荧光成像健康和龋坏的离体人牙；B为健康组和疾病组牙齿荧光强度数据输出图像；
图7为本发明的口腔喷剂荧光成像和光动力杀死体外龋病生物膜图。
具体实施方式
应该指出，以下详细说明都是例示性的，旨在对本申请提供进一步的说明；除非另有指明，本申请使用的所有技术和科学术语具有与本申请所属技术领域的普通技术人员通常理解的相同含义。
需要注意的是，这里所使用的术语仅是为了描述具体实施方式，而非意图限制根据本申请的示例性实施方式；如在这里所使用的，除非上下文另外明确指出，否则单数形式也意图包括复数形式，此外，还应当理解的是，当在本说明书中使用术语“包含”和/或“包括”时，其指明存在特征、步骤、操作、器件、组件和/或它们的组合。
具体地，本发明是通过如下所述的技术方案实现的：
本发明的第一种典型实施方式，提供了一种预防及快速诊断和治疗龋病的口腔喷剂的制备方法，将分子探针[4-(2-{4-[双-(4-甲氧基-苯基)-氨基]-苯基}-乙烯基)-1-甲基-吡啶鎓]+PF6 -(MeOTpy)和木糖醇溶于含有二甲基亚砜(DMSO)的磷酸盐(PBS)缓冲溶液中，即得预防及快速诊断和治疗龋病的口腔喷剂；
所述分子探针的结构式为：
在一种或多种实施方式中，所述MeOTpy的合成方法，包括如下步骤：
(1)将4-[双(4-甲氧基苯基)氨基]苯甲醛和1,4-碘化二甲基吡啶溶于乙醇中，在氮气保护下80℃回流9～11小时，反应完成后，冷却至室温，，洗涤即得到中间体Z；
(2)将中间体Z溶解于丙酮中，后加入饱和KPF6溶液，室温下搅拌1～2h，反应完成后，除去溶剂丙酮，过滤得到棕色固体，洗涤、干燥、纯化后得到MeOTpy；
优选的，步骤(1)4-[双(4-甲氧基苯基)氨基]苯甲醛和1,4-碘化二甲基吡啶溶于乙醇的摩尔比为：5:6。
优选的，步骤(1)乙醇的量为反应物当量的100～200倍。
优选的，步骤(1)中加入3～5滴哌啶作为催化剂。
优选的，步骤(1)中洗涤的方式为倒入大量乙醚中，后进行抽滤得到中间体Z。
优选的，步骤(2)中丙酮的量为反应物当量的50～100倍。
优选的，步骤(2)中间体Z与KPF6的摩尔比为：1：10～1：100。
优选的，步骤(2)饱和KPF6溶液的质量分数为20.9％。
优选的，步骤(2)中除去溶剂丙酮的方式为真空旋转，压力为0.01～0.1MPa，温度为30～35℃。
优选的，步骤(2)中洗涤的条件为用水洗涤3～5次。
优选的，步骤(2)中干燥的条件为在真空干燥箱中干燥温度为40～60℃，压强为0.01～0.1MPa。
优选的，步骤(2)中纯化的方法为：采用硅胶GF254薄层层析法纯化，洗脱溶剂为体积比为10:1的二氯甲烷/甲醇。
在一种或多种实施方式中，所述PBS缓冲溶液含有体积分数为0.001-0.1％二甲基亚砜，优选为0.1％。
在一种或多种实施方式中，所述MeOTpy和木糖醇的质量比为：43:2000。
在一种或多种实施方式中，所述PBS缓冲溶液的浓度为0.01mol/L，pH为7.4。
在一种或多种实施方式中，所述MeOTpy与PBS缓冲溶液的质量比是1：28000。
本发明的第二种典型实施方式，提供由上述口腔喷剂的制备方法制备的口腔喷剂。
为了使得本领域技术人员能够更加清楚地了解本发明的技术方案，以下将结合具体的实施例详细说明本发明的技术方案。
实施例1
MeOTpy的合成和表征：
具体的合成步骤如下：
将0.333g 4-[双(4-甲氧基苯基)氨基]苯甲醛和0.129g 1,4-碘化二甲基吡啶溶解在20ml乙醇中，然后加入4滴哌啶，在氮气保护下80℃回流搅拌10小时，反应完成后，将混合物冷却至室温，然后倒入大量乙醚中，抽滤得到中间体Z。
将0.2g中间体Z重新溶解在丙酮中，然后向混合物中加入10ml饱和KPF6(饱和溶液的质量分数为：20.9％)溶液，在室温下搅拌1小时，反应完成后，在压力为0.05MPa，温度为35℃时，旋蒸除去丙酮，过滤得到棕色固体，用水洗涤并在真空干燥箱中干燥，干燥的温度为50℃，压强为0.05MPa，利用硅胶GF254薄层层析法纯化，使用体积比为10:1的二氯甲烷/甲醇作为洗脱溶剂，得到棕色产物MeOTpy(67.4％)。如图1所示，通过质谱表征MeOTpy，HRMS(ESI)m/z calcd.for C28H27N2O2，[M-PF6]+calculated 423.21，found 423.20。
实施例2
本实施例探究了实施例1中制备的MeOTpy对致病菌S.mutans的快速荧光成像。
S.mutans在固体琼脂板中37℃培养8h，大肠杆菌(E.coli)在液体培养基LB中以100rpm的振摇速度恢复10h，然后转移到固体琼脂板上，在37℃下生长8h。利用无菌棉签收集两种细菌并分别在0.85％ NaCl溶液中稀释至2×108CFU(OD600＝0.257)。将S.mutans与E.coli混合，在浓度为7μM的MeOTpy中孵育30S、5min、10min和20min后进行时间依赖性共聚焦荧光成像。
如图2所示，MeOTpy加入细菌培养液30S时就可以点亮S.mutans，然而对E.coli几乎没有荧光响应。
利用荧光光谱仪器记录了MeOTpy对S.mutans的荧光响应动力学，图3显示，MeOTpy与S.mutans混合60S后荧光强度达到一个平台，因此，MeOTpy可以快速成像S.mutans。
实施例3
本实施探究了实施例1中制备的MeOTpy光动力杀菌性能。
DCFH用于检测在白光照射(10mW/cm2)下MeOTpy在生理条件下(PBS缓冲液，0.01M，pH为7.4)的活性氧(ROS)生成，具体的，DCFH与ROS反应后荧光会增强。将1μL MeOTpy母液(在DMSO中浓度为7mM)和1μL DCFH母液(1.0mM，10μL)添加到PBS缓冲液(989μL)中，记录DCFH在523nm处的荧光强度随白光照射时间的变化(488nm激发)。如图4所示，MeOTpy在白光照射下具有强的生成ROS的能力。
实施例4
本实施例探究了实施例1中制备的MeOTpy光动力杀死致病菌S.mutans。
使用传统的平板计数方法探究MeOTpy的抗菌性能，用不同浓度(0μM、1μM、3μM、5μM、7μM)的MeOTpy对主要的龋病致病S.mutans和一种共生菌大肠杆菌E.coli(107CFU/mL)在37℃、0.85％NaCl中孵育1min，后在白光下或黑暗中处理细菌15min，将处理过的细菌悬浮液连续稀释1000倍(104CFU/mL)，然后将100μL稀释的细菌铺展到固体琼脂板上，将处理后的细菌在37℃培养12h，计数每组形成的菌落数以评价抗菌效果。
如图5所示，在没有MeOTpy处理的情况下，S.mutans和E.coli在黑暗或光照条件下都能在琼脂平板上正常地生长和繁殖。MeOTpy对所有细菌的暗毒性均不明显，而在MeOTpy和白光照射下，S.mutans能够被有效杀死，这说明MeOTpy在有效光动力杀死致病菌的同时又能够避免杀死其他共生菌。
实施例5
口腔喷剂的制备：将实施例1中制备的0.0057g MeOTpy和20g木糖醇溶于1L含有质量分数为0.1％DMSO的0.01M、pH为7.4的PBS缓冲溶液，获得口腔喷剂。
实施例6
本实施例利用实施例5中制备的口腔喷剂检验离体牙齿中的龋病生物膜：
收集健康和龋坏的离体牙齿，将实施例5种制备的口腔喷剂喷洒在离体牙齿表面，后对健康和龋坏的离体牙齿进行荧光成像。如图6所示，口腔喷剂可以对牙齿进行荧光成像，龋病组的荧光强度高于健康组，并且能够在宏观上指示龋坏潜在位点，因此，口腔喷剂可以评估龋齿程度。
实施例7
利用实施例5中制备的口腔喷剂杀灭龋病细菌生物膜：
收集重度龋齿患儿牙菌斑，接种于健康牙釉质表面，高糖培养形成细菌生物膜。利用实施例5中制备的口腔喷剂分别在黑暗和光照条件下处理细菌生物膜15min。使用SYTO60对细菌生物膜中所有细菌荧光染色，利用SYTOX-Green对细菌生物膜中的死细菌进行染色。如图7所示，共聚焦荧光成像结果表明，口腔喷剂在黑暗条件下处理的细菌生物膜仅观察到SYTO60的橙色荧光，但是，口腔喷剂在光照下处理的细菌生物膜观察到SYTO60的橙色荧光和SYTOX-Green的绿色荧光，表明口腔喷剂可以有效杀灭细菌生物膜。
因此，本发明提供的口腔喷剂能够快速有效的实时成像和杀死口腔细菌生物膜，防治龋病，清新口气。同时又避免了杀死口腔中的一些共生菌，降低龋病诊疗费用，简化了诊疗过程，在龋病诊断、治疗中显示出良好的前景。
以上所述仅为本发明的优选实施例而已，并不用于限制本发明，对于本领域的技术人员来说，本发明可以有各种更改和变化。凡在本发明的精神和原则之内，所作的任何修改、等同替换、改进等，均应包含在本发明的保护范围之内。
Claims (10)
1.一种预防及快速诊断和治疗龋病的口腔喷剂的制备方法，其特征在于，所述方法为：将分子探针[4-(2-{4-[双-(4-甲氧基-苯基)-氨基]-苯基}-乙烯基)-1-甲基-吡啶鎓]+PF6 -和木糖醇溶于含有二甲基亚砜的磷酸盐缓冲溶液中，即得预防及快速诊断和治疗龋病的口腔喷剂；
所述分子探针的结构式为：
2.如权利要求1所述的制备方法，其特征在于，所述MeOTpy的合成方法，包括如下步骤：
(1)将4-[双(4-甲氧基苯基)氨基]苯甲醛和1,4-碘化二甲基吡啶溶于乙醇中，在氮气保护下80℃回流9～11小时，反应完成后，冷却至室温，，洗涤即得到中间体Z；
(2)将中间体Z溶解于丙酮中，后加入饱和KPF6溶液，室温下搅拌1～2h，反应完成后，除去溶剂丙酮，过滤得到棕色固体，洗涤、干燥、纯化后得到MeOTpy；
3.如权利要求2所述的制备方法，其特征在于，所述步骤(1)4-[双(4-甲氧基苯基)氨基]苯甲醛和1,4-碘化二甲基吡啶溶于乙醇的摩尔比为：5:6；
或，步骤(1)乙醇的量为反应物当量的100～200倍；
或，步骤(1)中加入3～5滴哌啶作为催化剂。
4.如权利要求2所述的制备方法，其特征在于，步骤(1)中洗涤的方式为倒入大量乙醚中，后进行抽滤得到中间体Z。
5.如权利要求2所述的制备方法，其特征在于，所述步骤(2)中丙酮的量为反应物当量的50～100倍；
或，步骤(2)中间体Z与KPF6的摩尔比为：1：10～1：100；
或，步骤(2)饱和KPF6溶液的质量分数为20.9％。
6.如权利要求2所述的制备方法，其特征在于，步骤(2)中除去溶剂丙酮的方式为真空旋转，压力为0.01～0.1MPa，温度为30～35℃；
或，步骤(2)中洗涤的条件为用水洗涤3～5次；
或，步骤(2)中干燥的条件为在真空干燥箱中干燥温度为40～60℃，压强为0.01～0.1MPa。
7.如权利要求2所述的制备方法，其特征在于，步骤(2)中纯化的方法为：采用硅胶GF254薄层层析法纯化，洗脱溶剂为体积比为10:1的二氯甲烷/甲醇。
8.如权利要求1所述的制备方法，其特征在于，所述PBS缓冲溶液含有体积分数为0.001-0.1％二甲基亚砜，优选为0.1％；
或，所述MeOTpy和木糖醇的质量比为：43:2000；
或，所述PBS缓冲溶液的浓度为0.01mol/L，pH为7.4。
9.如权利要求1所述的制备方法，其特征在于，所述MeOTpy与PBS缓冲溶液的质量比是1：280000。
10.权利要求1～9项任一项所述的制备方法制备的口腔喷剂。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310267812.1A CN116440077A (zh) | 2023-03-15 | 2023-03-15 | 一种预防及快速诊断和治疗龋病的口腔喷剂及其制备方法 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310267812.1A CN116440077A (zh) | 2023-03-15 | 2023-03-15 | 一种预防及快速诊断和治疗龋病的口腔喷剂及其制备方法 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN116440077A true CN116440077A (zh) | 2023-07-18 |
Family
ID=87129313
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202310267812.1A Pending CN116440077A (zh) | 2023-03-15 | 2023-03-15 | 一种预防及快速诊断和治疗龋病的口腔喷剂及其制备方法 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116440077A (zh) |
Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US8647119B1 (en) * | 2006-04-18 | 2014-02-11 | President And Fellows Of Harvard College | Methods and kits with fluorescent probes for caries detection |
| CN104622871A (zh) * | 2015-01-21 | 2015-05-20 | 四川大学 | 一种恶唑酮类化合物作为变异链球菌生物膜抑制剂的应用 |
| WO2015105294A1 (ko) * | 2014-01-07 | 2015-07-16 | 경북대학교 산학협력단 | 5탄당 및 자일리톨을 유효성분으로 포함하는 구강 세균 부착 억제용 조성물 |
| CN106588911A (zh) * | 2016-12-14 | 2017-04-26 | 山东大学 | 一种抗变形链球菌的新型噻唑类化合物xqh‑3‑6及其应用 |
| CN108379256A (zh) * | 2018-04-23 | 2018-08-10 | 山东大学 | 一种用于预防和辅助治疗牙周炎的口腔喷剂及其制备方法 |
| CN108434174A (zh) * | 2018-04-23 | 2018-08-24 | 山东大学 | 一种用于辅助龋齿治疗及预防龋齿的口腔喷剂及其制备方法 |
| CN109503510A (zh) * | 2019-01-03 | 2019-03-22 | 山东大学 | 一种防龋抗菌的噻唑类化合物及其制备方法 |
| CN110790698A (zh) * | 2019-08-23 | 2020-02-14 | 深圳大学 | 一种深红/近红外多功能聚集诱导发光材料及其制备方法和应用 |
| CN113171470A (zh) * | 2020-01-24 | 2021-07-27 | 香港科技大学 | 聚集诱导发光分子—噬菌体生物偶联物 |
-
2023
- 2023-03-15 CN CN202310267812.1A patent/CN116440077A/zh active Pending
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US8647119B1 (en) * | 2006-04-18 | 2014-02-11 | President And Fellows Of Harvard College | Methods and kits with fluorescent probes for caries detection |
| WO2015105294A1 (ko) * | 2014-01-07 | 2015-07-16 | 경북대학교 산학협력단 | 5탄당 및 자일리톨을 유효성분으로 포함하는 구강 세균 부착 억제용 조성물 |
| CN104622871A (zh) * | 2015-01-21 | 2015-05-20 | 四川大学 | 一种恶唑酮类化合物作为变异链球菌生物膜抑制剂的应用 |
| CN106588911A (zh) * | 2016-12-14 | 2017-04-26 | 山东大学 | 一种抗变形链球菌的新型噻唑类化合物xqh‑3‑6及其应用 |
| CN108379256A (zh) * | 2018-04-23 | 2018-08-10 | 山东大学 | 一种用于预防和辅助治疗牙周炎的口腔喷剂及其制备方法 |
| CN108434174A (zh) * | 2018-04-23 | 2018-08-24 | 山东大学 | 一种用于辅助龋齿治疗及预防龋齿的口腔喷剂及其制备方法 |
| CN109503510A (zh) * | 2019-01-03 | 2019-03-22 | 山东大学 | 一种防龋抗菌的噻唑类化合物及其制备方法 |
| CN110790698A (zh) * | 2019-08-23 | 2020-02-14 | 深圳大学 | 一种深红/近红外多功能聚集诱导发光材料及其制备方法和应用 |
| CN113171470A (zh) * | 2020-01-24 | 2021-07-27 | 香港科技大学 | 聚集诱导发光分子—噬菌体生物偶联物 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Lu et al. | Redox/pH dual-controlled release of chlorhexidine and silver ions from biodegradable mesoporous silica nanoparticles against oral biofilms | |
| Shrestha et al. | Antibiofilm efficacy of photosensitizer-functionalized bioactive nanoparticles on multispecies biofilm | |
| Bukhari et al. | Novel endodontic disinfection approach using catalytic nanoparticles | |
| Sun et al. | Nanoparticles having amphiphilic silane containing Chlorin e6 with strong anti-biofilm activity against periodontitis-related pathogens | |
| CN110790698A (zh) | 一种深红/近红外多功能聚集诱导发光材料及其制备方法和应用 | |
| Albuquerque et al. | Antimicrobial effects of novel triple antibiotic paste–mimic scaffolds on Actinomyces naeslundii biofilm | |
| Ostadhossein et al. | Dual purpose hafnium oxide nanoparticles offer imaging Streptococcus mutans dental biofilm and fight it in vivo via a drug free approach | |
| Tang et al. | Fucoidan-derived carbon dots against Enterococcus faecalis biofilm and infected dentinal tubules for the treatment of persistent endodontic infections | |
| Jurič et al. | Antimicrobial efficacy of photodynamic therapy, Nd: YAG laser and QMiX solution against Enterococcus faecalis biofilm | |
| Daood et al. | Effect of a novel quaternary ammonium silane cavity disinfectant on cariogenic biofilm formation | |
| Balto et al. | The efficacy of Salvadora persica extract in the elimination of the intracanal smear layer: A SEM study | |
| Shrestha et al. | Bioactivity of photoactivated functionalized nanoparticles assessed in lipopolysaccharide-contaminated root canals in vivo | |
| Shui et al. | Inhibitory effects of sodium new houttuyfonate on growth and biofilm formation of Streptococcus mutans | |
| Li et al. | Multifunctional magnesium organic framework-based photothermal and pH dual-responsive mouthguard for caries prevention and tooth self-healing promotion | |
| CN116440077A (zh) | 一种预防及快速诊断和治疗龋病的口腔喷剂及其制备方法 | |
| Ballal et al. | Biological and chemical properties of 2-in-1 calcium-chelating and antibacterial root canal irrigants | |
| CN113645941A (zh) | 口腔护理产品 | |
| Hasegawa et al. | A horizontal sequential cutting method to estimate the effectiveness of dentin disinfection by using confocal laser scanning microscopy | |
| Nagarathinam et al. | Effect of triple antibiotic loaded apatitic nanocarriers on Enterococcus faecalis biofilm–an In vitro study | |
| CN116726194A (zh) | 一种卟啉-抗生素超分子纳米颗粒和制备方法及其应用 | |
| Wang et al. | A study on the use of phase transition lysozyme-loaded minocycline hydrochloride in the local treatment of chronic periodontitis | |
| Wyganowska-Swiatkowska et al. | The evaluation of enamel matrix derivative on subgingival microbial environment in non-surgical periodontal therapy | |
| Shi et al. | Photodynamic antimicrobial activity against S. aureus of metallic Iridium (III) complexes with Helicin pendant | |
| Fu et al. | The applications of carbon dots in oral health: A scoping review | |
| Liang et al. | Ebselen Optimized the Therapeutic Effects of Silver Nanoparticles for Periodontal Treatment |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |