WO2004093876A2 - Procede et composition pour reguler des pathogenes oraux - Google Patents

Procede et composition pour reguler des pathogenes oraux Download PDF

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Publication number
WO2004093876A2
WO2004093876A2 PCT/US2004/008616 US2004008616W WO2004093876A2 WO 2004093876 A2 WO2004093876 A2 WO 2004093876A2 US 2004008616 W US2004008616 W US 2004008616W WO 2004093876 A2 WO2004093876 A2 WO 2004093876A2
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Prior art keywords
berberine
antibiotic
antimicrobial
administered
bromoberberine
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PCT/US2004/008616
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English (en)
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WO2004093876A3 (fr
Inventor
Christine D. Wu
Douglas Kinghorn
Sara Kate Roberts
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The Board Of Trustees Of The University Of Illinois
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Priority to US10/551,639 priority Critical patent/US20070098649A1/en
Publication of WO2004093876A2 publication Critical patent/WO2004093876A2/fr
Publication of WO2004093876A3 publication Critical patent/WO2004093876A3/fr

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/13—Amines
    • A61K31/155—Amidines (), e.g. guanidine (H2N—C(=NH)—NH2), isourea (N=C(OH)—NH2), isothiourea (—N=C(SH)—NH2)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/47—Quinolines; Isoquinolines
    • A61K31/4738—Quinolines; Isoquinolines ortho- or peri-condensed with heterocyclic ring systems
    • A61K31/4741—Quinolines; Isoquinolines ortho- or peri-condensed with heterocyclic ring systems condensed with ring systems having oxygen as a ring hetero atom, e.g. tubocuraran derivatives, noscapine, bicuculline
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/65—Tetracyclines
    • 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
    • A61P31/04—Antibacterial agents

Definitions

  • the present invention relates to the control of oral and other human pathogens, and to the treatment of diseases and conditions that directly or indirectly result from such pathogens, by administering (a) berberine and (b) an antimicrobial or an antibiotic. More particularly, the present invention relates to a method of potentiating the effects of. an antimicrobial or an antibiotic in a mammal by administration of therapeutically effective amounts of both berberine and an antimicrobial or an antibiotic.
  • the composition and method of the present invention permit a reduction in the antimicrobial or . antibiotic dose and in the berberine dose while providing a desired therapeutic effect, thus reducing adverse side effects .
  • Dental plaque frequently is associated with oral diseases, including dental caries and periodontal disease (i.e., gum disease). Plaque control can be achieved by mechanical (e.g., brushing and flossing) or chemical means.
  • antimicrobial agents have been incorporated into oral hygiene products, such as toothpastes and mouth rinses, to control plaque and gingivitis/periodontal disease .
  • CHX antimicrobial chlorhexidine
  • TCX chlorhexidine
  • Tetracyclines are conventional antibiotics, and routinely are used in the treatment, of periodontal disease because of their unique ability to accumulate in the periodontal pocket area.
  • Tetracycline antibiotics also have many adverse side effects, including gastrointestinal discomfort, nausea, vomiting, diarrhea, and permanent tooth discoloration in children up to eight years old.
  • Berberine is a plant alkaloid having a long history of medicinal use in both Ayurvedic and Chinese medicine. Berberine is present in the roots, rhizomes, and stem bark of Hydras tis canaden- sis (goldenseal) , Coptis ch ⁇ nensis (coptis or goldenthread) , Berberis aquifolium (Oregon grape) , Berberis vulgaris (barberry) , and Berberis aristata (tree tumeric) .
  • Berberine is 5, 6-dihydro-9 , 10- dimethoxybenzo [g] -1, 3-benzodioxolo [5, 6-a] quinolizin- ium, and as used 'herein, the term "berberine” includes free berberine and salts thereof.
  • Goldenseal is a major phytomedicine sold 5 in the United States and is one of the top five herbal supplements in the worldwide market . Golden- seal is used in various dietary supplements for the purpose of enhancing the immune response of the body. Goldenseal-containing oral hygiene products
  • Berberine extracts and decoctions demonstrate significant antimicrobial activity against a
  • antimicrobials and antibiotics exhibit several adverse side effects, including tooth discoloration, bad taste, and nausea, but they are routinely used in the treatment
  • a combination treatment utilizing berberine • and a conventional antimicrobial or antibiotic agent allows for decreased concentrations of both agents, which reduces the occurrence or sever-
  • the present invention is directed to the discovery that an antimicrobial or antibiotic and berberine, when administered in combination, perform synergistically. Therefore, the amount of antimicrobial or an i- biotic and berberine used to control the oral pathogens can be reduced, with a concomitant reduction or elimination of adverse side effects.
  • the combined use of an antimicrobial or antibiotic and a berberine also can be used to synergistically con- trol nonoral human • pathogens .
  • berberine potentiates the antimicrobial effects of an antimicrobial or antibiotic in the treatment of oral and other human, pathogens.
  • antibiotic an antimicrobial or antibiotic
  • the reduced amount of antibiotic and berberine re- quired to provide a desired antimicrobial effect reduces the severity of various adverse side effects associated with antibiotic and berberine treatment.
  • one aspect of the present in- vention is to provide a composition comprising (a) an antibiotic, e.g., CHX,- tetracycline, or doxy- cycline, and (b) berberine, or a salt, derivative, or prodrug thereof, for use in controlling oral and other human pathogens .
  • the present invention therefore, provides a composition and method for improved control of oral pathogens, and treatment of diseases and condi- • tions caused by oral pathogens, like gingivitis and dental caries.
  • the present invention is directed to compositions containing berberine and an antibiotic, and to methods of using the composition to prevent and/or treat diseases and conditions of the mouth, teeth, and gums. More particularly, the present invention is directed to compositions • containing berberine and CHX or a tetracycline, and to use of the compositions in methods of treating diseases and conditions of the mouth, teeth, and gums .
  • Still another aspect of the present inven- tion is to provide a method and composition for controlling oral and nonoral human pathogens, including antibiotic-resistant pathogens.
  • Another aspect of the present invention is to provide an article of manufacture for human use comprising (a) a package insert, (b) a container, and either (cl) a packaged- composition comprising an antibiotic and berberine or (c2) a packaged composition comprising an antibiotic and a packaged composition comprising berberine.
  • Appendix A discloses test results relating to administration of a berberine and an antibiotic to control oral pathogens.
  • the tests show that administration of berberine synergistically potentiates the antimicrobial effect of an antibiotic, and allows a reduction in the dose of both the antibiotic and berberine, while providing an antimicrobial effect equivalent to using a higher dose of antibiotic or berberine used alone.
  • Such a synergistic coadministration provides an effective treatment, reduces adverse side effects associated with antibiotic or berberine administration, and addresses the problem of bacterial resistance to antibiotics .
  • the present invention therefore, provides methods of synergistically potentiating the antimicrobial properties of an antibiotic and berberine in the treatment of oral and other human pathogens.
  • the invention also provides pharmaceutical compositions comprising an antibiotic and berberine.
  • the present invention also provides articles of manufacture comprising an antibiotic and berberine, packaged separately or together, and an insert having instructions for using these active agents in the control of oral and other human pathogens, and the treatment of diseases and conditions caused by such pathogens .
  • the methods described herein benefit from the use of an antibiotic and berberine in the treatment ' and management of oral and other human pathogens .
  • the antibiotic and berberine can be administered simultaneously or sequentially to achieve the desired synergistic antimicrobial effect.
  • treatment includes preventing, controlling, lowering, or eliminating the concentration of oral and other human pathogens.
  • treatment includes both medical therapeutic and/or prophylactic administration, as appropriate.
  • container means any receptacle and optional closure therefor suitable for storing, shipping, dispensing, and/or handling an article of manufacture .
  • the term "insert” means information accompanying an article of manufacture that provides a description of how to administer the article, along with the safety and efficacy data required to allow the physician, pharmacist, or user to make an informed decision regarding use of the article .
  • the package insert generally is regarded as the "label” for an article.
  • prodrug means compounds that transform rapidly in vivo to a compound useful in the invention, for example, by hydrolysis. A thorough discussion of prodrugs is provided in Higuchi et al . , Prodrugs as Novel Delivery Systems, Vol . 14 , of the A.C.S.D. Symposium Series, and in Roche (ed.) Bioreversible Carriers in Drug Design, American Pharmaceutical Association and Pergamon Press, 1987.
  • an antibiotic and a berberine are present in a composition, or are administered in combination, in a weight ratio of antibiotic-to- berberine of about 0.01:1 to about 100:1, and preferably about 0.01:1 to about 10:1.
  • This ratio de- pends upon the type and identity of antibiotic and berberine being used.
  • the ratio of antibiotic-to- berberine that is administered is dependent upon the particular antibiotic and berberine used, and the origin and severity of the condition being treated. This ratio can be readily determined by a person skilled in the art to achieve the desired control of an oral or other human pathogen.
  • berberine When berberine is used in combination with an antibiotic, such as CHX or a tetracycline, like doxycycline, a synergistic antimicrobial growth inhibition effect is observed. Such a combination treatment results in, a lower minimum inhibitory concentration (MIC) of either active agent.
  • an antibiotic such as CHX or a tetracycline, like doxycycline
  • Antibiotics most commonly are used in dental practice as prophylactic agents for preventa- tive management of endocarditis.
  • antibiotics are used therapeutically in cases where infections of oral hard and soft tissues, such as teeth and gingival, cannot be controlled by local debridement and can spread to distant organs, and, therefore, require supplemental therapy.
  • Tetracycline antibiotics have a broad spectrum of activity, can be used by many routes of administration, such as oral administration and sustained delivery systems, and are widely used. Tetracyclines inhibit protein synthesis by combining the small (30S) subunit of the riboso e and inhibiting the binding of aminocyl-tRNA molecules to the • ribosomal A site. Antibiotics generally are considered bacteriostatic . Reports of bacterial resistance to tetracyclines, both in vi tro and in vivo, are increasing. However, antibiotics still are useful, and are routinely used in periodontal treatment. Tetracyclines concentrate in periodontal pockets at inhibitory levels 2 to 4 times higher than in the blood, strongly bind to root surfaces, and can be released in active form over extended time periods.
  • Chlorhexidine is a bisbiguanide biocide, and is marketed in the, form of different salts . Chlorhexidine glucpnate is one of the most common of these salts. CHX-containing compositions have been used as topical disinfectants since the middle 1970s. Because chlorhexidine in the salt form is cationic, it readily binds to negatively charged cell walls. CHX targets the cytoplasmic membrane of bacteria resulting in the loss of structural organization and integrity. Subsequently, coagulation and precipitation of cyto- plasmic constituents occurs.
  • CHX can be employed as an adjunct to other preventative measures.
  • chlorhexidine is used for postoperative rinsings and as an adjunct to mechanical debriment .
  • CHX compositions also are used for nonspecific plaque control, as well as against specific bacteria associated with periodontal disease and caries and in subgingival irrigation.
  • CHX usually is administered from rinsing solutions (concentrations, typically about 1-2%) , and slow release devices, such as varnishes, fibers, and slab-like sustained delivery devices.
  • CHX also can be incorporated into chewing gum, toothpastes, and toothbrushes, or in an oral rinse or in a disin- fectant for dentures in patients with candidial infections .
  • CHX has a low level of toxicity both locally and systemically and shows no permanent retention in the body.
  • the antibiotic incorporated into a present composition or utilized in a present .method is not ' limited to CHX. Any other antibiotic useful in the . treatment of oral and other human pathogens, like the tetracyclines, can be used in the present composition and methods.
  • useful antibiotics include, but are not limited to, chlorhexidine, chlorhexidine dihydrochloride, chlorhexidine diacetate salt hydrate, chlorhexidine digluconate, alexidine, alexidine dihydrochloride, tetracycline, tetracycline hydrochloride, doxycycline, doxycycline hydrochloride, and mixtures thereof.
  • the second active agent in a present composition and method is berberine.
  • berberine markedly improves cardiac performance in patients with heart conditions when administered at 0.02 mg/kg/min (59 nmol/kg/min) for 30 minutes, followed by 0.2 mg/kg/min (0.59 mol/kg/min) for an additional 30 minutes.
  • berberine administration to heart failure patients can result in ventricular tachycardia in some subjects.
  • Berberine also has an anesthetic effect when injected s.c, 5 and berberine produced parapsympatholytic and anesthetic effects when applied to the eyes.
  • Berberine sulfate is absorbed through human skin. Following oral administration, berberine is absorbed slowly, requiring four hours to
  • mice In mice, the oral LD 50 dose of berberine is
  • the s.c. LD 50 dose is 18 mg/kg (0.054 mmol/kg), and the i.p. LD dose is greater than 500 mg/kg (>1.49 mmol/kg (>2.31 mmol/- kg) orally.
  • the s.c. LD 50 dose is 100 mg/kg (0.30 mmol/kg) .
  • the present invention is directed to the use of berberine, a natural compound found in goldenseal, in combination with an- antibiotic, e.g., CHX or a tetracycline, in the control of oral and other human pathogens, and diseases and conditions directly or indirectly caused by such pathogens.
  • the invention is not limited to the use of a par- ticular berberine, but extends to free berberine, all pharmaceutically acceptable salts of berberine, derivatives of berberine, and prodrugs of berberine.
  • berberines and berberine derivatives useful in the present invention include, but are not limited to, free berberine, berberine chloride, berberine bisulfate, berberine hemisul- fate, a berberine prodrug, a berberine derivative listed in the following Table 1, and mixtures thereof.
  • the present invention is the first disclosure of a combined treatment using berberine- and an antibiotic, like chlorohexadine or tetracycline, to treat gingivitis and other diseases and conditions of the mouth, teeth, and gums.
  • the present invention also can be used to treat diseases and conditions directly or indirectly caused by nonoral human pathogens.
  • berberine is administered in combination with an antibiotic, it has been found that the dose of both the antibiotic and berberine can be reduced by a factor of up to four to achieve the same bacterial growth inhibition ob- served using either • an antibiotic or berberine alone .
  • the present invention demonstrates that berberine chloride, when administered with CHX or an antibiotic, exerts synergistic growth inhibitory activities. '
  • oral hygiene products and toothpastes containing goldenseal have become widely available in the U.S.
  • Antibiotics are used ' in the treatment of periodontal disease.
  • the present invention allows the use of reduced concentra- tions of CHX and a tetracycline in oral hygiene/- therapeutic products to achieve a desired antimicrobial effect, thus minimizing the occurrence of adverse effects attributed to antibiotics and berberine and addressing the problem of pathogen resis- tance to antibiotics.
  • berberine hydrochloride When berberine hydrochloride was administered in combination with chlorhexidine gluconate (CHX) or a tetracycline antibiotic (e.g., tetra- cycline hydrochloride or doxycycline hydrochloride) , a synergistic microbial growth inhibition effect was observed.
  • CHX chlorhexidine gluconate
  • a tetracycline antibiotic e.g., tetra- cycline hydrochloride or doxycycline hydrochloride
  • Berberine chloride purchased from Sigma (St. Louis, MO), was tested to assess whether a different antimicrobial activity existed between a compound isolated from goldenseal and commercially available berberine chloride. The antimicrobial investigations showed no difference in antimicrobial activity between the two sources of barbering chloride .
  • a combination treatment using berberine hydrochloride and a conventional antimicrobial agent allows for decreased concentrations of both agents in the treatment of mouth, teeth, and gum conditions and diseases, and reduces the occurrence of adverse side effects. This is an important finding because of the increase in resistance to conventional antibacterial treatments.
  • the present invention therefore, provides a new composition and method for enhanced treatment of mouth, teeth, and gum diseases and conditions, and for the treatment of other diseases and conditions caused, directly or indirectly / by pathogens controlled by berberine and/or an antibiotic .
  • pathogens controlled by berberine and/or an antibiotic are two nonoral, human pathogens.
  • Berberrubine structurally similar to berberine, also was evaluated for antimicrobial activity against S. wutans, A . actinomycetecomi tan , and P. gingivalis .
  • the test results indicated that berberrubine was not antimicrobial against A . actinomycetecomi tans at concentrations less than 300 ⁇ g ml "1 .
  • the test results also suggest that berberrubine may be antimicrobial against S . mutans at concentrations greater than 300 ⁇ g ml -1 because, at lower concentrations, growth was hindered, but not inhibited.
  • Berberrubine demonstrated antimicrobial activity against P . gingivalis , at concentrations comparable to berberine (i.e., MIC 17 ⁇ g ml "1 ) .
  • test results clearly demonstrate that the antimicrobial effects of an antibiotic and berberine on oral and other human pathogens are synergistically potentiated when the agents are co- administered, sequentially or simultaneously. This is an important clinical finding because the amount of antibiotic and berberine required to achieve a desired antimicrobial effect can be substantially reduced, which in turn substantially reduces adverse effects attributed to antibiotic and berberine use.
  • Berberine combined with an antibiotic therefore, can be used to potentiate the antimicro- bial action of an antibiotic in the treatment of mouth, teeth, and gum diseases, and other diseases and conditions directly or indirectly caused by human pathogens.
  • the test results show that, when combined with berberine administration, antibiotics produce significant antimicrobial control using a much lower dose of antibiotic and berberine, and, therefore, ' bacterial resistance to the antibiotic and berberine is reduced.
  • the tests and data show that a combination of an antibiotic and berberine can be administered to mammals in methods of controlling oral and other human pathogens, and treating pathogen-mediated diseases and conditions, especially of the mouth, teeth, and gums.
  • the antibiotic and berberine i.e., active ingredients
  • the active agents typically- are present in such a composition in an amount of about 0.1% to about 75% by weight, either alone or in combination.
  • compositions containing the active agents, i.e., antibiotic and berberine, of the present invention are suitable for administra- tion to humans or other mammals.
  • the pharmaceutical compositions are sterile, and contain no toxic, carcinogenic, or mutagenic compounds that cause an adverse 'reaction when administered.
  • Administration of the pharmaceutical composition can be performed before, during, or after the onset of a disease or condition, e.g., gingivitis, endocarditis, or plaque formation.
  • the method of the present invention can be accomplished using the active agents as described above individually as the neat compounds, or as a physiologically acceptable salt, solvate, derivative, or prodrug thereof. .
  • the active agents can be administered neat, or as a pharmaceutical composition containing either or both entities.
  • the active agents can be administered by any suitable route, .for example, by oral, topical, bucca ' l, sublingual, nasal, percutaneous, i.e., fransdermal, or parenteral (including intravenous, intramuscular, subcutaneous, and intracoronary) administration.
  • Parenteral administration can be accomplished using a needle and syringe, or using a high pressure technique, like POWDERJECTTM .
  • the active agents are administered orally, buccally, sublingually, or topically.
  • compositions include those wherein the active ingredients are administered in an effective amount to achieve their intended purpose. More specifically, a "therapeutically effective amount” means an amount effective to prevent development of, to eliminate, or to alleviate a mouth, teeth, gum, or other disease or condition. Determination of a therapeutically effective amount is well within the capability of those skilled in the art, especially in light of the detailed disclosure provided herein.
  • a “therapeutically effective dose” refers to that amount of the active agents that results in achieving the desired effect. Toxicity and therapeutic efficacy of such active agents can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., determining the LD 50 (the dose lethal to 50% of the population) and the ED 50 (the dose therapeutically effective in 50% of the population) . The dose ratio between ' toxic and therapeutic effects is the therapeutic index, which is expressed as the ratio between LD 50 and ED 50 . A high therapeutic index is preferred. The data obtained from such data can be used in formulating a range of dosage for use in humans . The dosage of the active agents preferably lies within a range of circulating concentrations that include the ED 50 with little or no toxicity. The dosage can vary within this range depending upon the dosage form employed, and the route of administration utilized.
  • the exact formulation, route of adminis- tration, and dosage is determined by an individual or physician in view of the individual's condition. Dosage amount and interval can be adjusted individ- . ually to provide levels of the active agents that are sufficient to maintain therapeutic or prophyl- actic effects.
  • the amount of pharmaceutical composition administered is dependent on the subject being treated, on the subject's weight/ the severity of the affliction, the manner of administration, and the judgment of the prescribing physician.
  • dosages of an antibiotic and berberine for administration to a human in the curative or prophylactic treatment of oral and other human pathogens, generally are about 10 to about 200 mg daily for an average adult patient (70 kg) , typically- divided into two to three doses per day or administered as 'a time-release formulation.
  • individual dosage forms contain about 0.1 to about 200 mg antibiotic and about 0.1 to about ' 200 mg berberine, in a suitable pharmaceutically acceptable vehicle or carrier, for administration in single or multiple doses, once or several times per day.
  • Dosages for intravenous, buccal, topical, or sublingual administration typically are about 0.1 to about 10 mg/kg per single dose as required.
  • the physician determines the actual dosing regimen which is most suitable for an individual patient, and the dosage varies with the age, .weight, and response of the particular patient.
  • the above dosages are exemplary of the average case, but there can be individual instances in which higher or lower dosages are merited, and such are within the scope of this invention.
  • the active agents of the present invention can be administered alone, or in admixture with a pharmaceutical carrier selected with regard to the intended route of administration and standard pharmaceutical practice.
  • Pharmaceutical compositions for use in accordance with the present invention thus can be . formulated in a conventional manner using one or more physiologically acceptable carriers comprising excipients and auxiliaries that facilitate processing of the active agents into preparations which can' be used pharmaceutically.
  • compositions can be manufactured in a conventional manner, e.g., by conventional mixing, dissolving, granulating, dragee-making, emulsifying, encapsulating,, entrapping, or lyophilizing processes. Proper formulation is dependent upon the route of administration chosen.
  • the composition typically is in the form of a tablet, capsule, powder, solution, or elixir.
  • the composition can additionally contain a solid carrier, such as gelatin or an adjuvant.
  • the tablet, capsule, and powder contain about 5% to about 95% of an active agent of the present invention, and preferably from about 25% to about 90% compound of the present invention.
  • a liquid carrier such as water, petroleum, or oils of. animal or plant origin
  • the liquid form of the composition can further contain physi- ological saline solution, dextrose or other saccharide solutions, or glycols.
  • the composition When administered in liquid form, the composition contains about 0.5% to about 90% by weight of active agents, and preferably about 1% to about 50% of an active agent.
  • the active agents can be readily combined with pharmaceutically acceptable carriers well known in the art. Such carriers enable the active agents to be formulated as tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions, and the like, for oral ingestion by a patient to be treated.
  • Pharmaceutical preparations for oral use can be obtained by adding the active agents with a solid excipient, optionally grinding the resulting mixture, and processing t e mixture of granules, after adding suitable auxiliaries, if desired, to obtain tablets or dragee cores. Suitable excipients include, for example, fillers and ' cellulose preparations. If desired, disintegrating agents can be added .
  • the active agents can be formulated for parenteral administration by injection, e.g., by bolus injection or continuous infusion.
  • Formulations for injection can be presented in unit dosage form, e.g., in ampules or in multidose containers, with an added preservative.
  • the compositions can take such forms as suspensions, solutions, or emulsions in oily or aqueous vehicles, and can contain formulatory agents such as suspending, stabilizing, and/or dispersing agents .
  • compositions for parenteral administration include aqueous solutions of the active agent in water-soluble form.
  • suspensions of the active agents can be prepared as appropriate oily 'injection suspensions.
  • Suitable lipophilic solvents or vehicles include fatty oils or synthetic fatty acid esters.
  • Aqueous injection suspensions can contain substances which increase the viscosity of the suspension.
  • the suspension also can contain suitable stabilizers or • agents that increase the solubility of the compounds and allow for the preparation of highly concentrated .solutions.
  • a present composition can be in .powder form for constitution with a suitable vehicle, e.g., sterile pyrogen-free water, before use.
  • the active agents also can be formulated as a depot preparation.
  • Such long-acting formulations can be administered by implantation (for example, subcutaneously or intramuscularly) or by intramuscular injection.
  • the active agents can be formulated with suitable polymeric or hydrophobic materials (for example, as an emulsion in an acceptable oil) or ion exchange resins, or as sparingly soluble derivatives, for example, as a sparingly soluble salt.
  • the active agents can be administered orally, buccally, or sublingually in the form of tablets containing excipients, such as starch or lactose, or in capsules or ovules, either alone or in admixture with excipients, or in the form of elixirs of suspensions containing flavoring or coloring agents.
  • excipients such as starch or lactose
  • capsules or ovules either alone or in admixture with excipients, or in the form of elixirs of suspensions containing flavoring or coloring agents.
  • Such liquid preparations can be prepared with pharmaceutically acceptable additives, such as suspending agents.
  • a compound also can be injected parenterally, for example, intravenously, intramuscularly, subcutaneously, or intracoronarily .
  • the compound is best used in the form of a sterile aqueous solution which can contain other substances, for example, salts, or monosaccharides , such as mannitol or glucose, to make the solution isotonic with blood.
  • the active ingredients are administered as a suitably acceptable formulation in accordance with normal • veterinary practice. The veterinarian can readily determine the dosing regimen and route of administration that is most appropriate for a particular animal .
  • an antibiotic is widely used in the treatment of several disease conditions of the mouth and gums, including gingivitis.
  • a major problem in the use of an antibiotic in an oral application is their adverse side effects.
  • the problem of bacterial resistance to an antibiotic also exists. It has been discovered that using berberine together with an antibiotic synergistically potentiates the antimicrobial action of the antibiotic, thereby allowing the administration of a reduced amount of antibiotic and berberine to achieve a desired result while addressing bacterial resistance associated with antibiotic and berberine use.
  • goldenseal is a relatively rare plant and only a small portion of commercially available goldenseal is derived from cultivated crops.
  • the present invention is important with respect to identifying the active components of goldenseal that are formulated into efficacious products, such as oral hygiene products.
  • Berberine derivatives or prodrugs also can be used in accord- ance with 'the present invention to overcome - availability problems associated with naturally occurring berberine and to reduce the costs associated with extracting berberine from plant sources .
  • the synergistic effect of the combined application of berber- ine and antibiotic also applies to other human pathogens and, therefore, the combination can be used in the treatment, of nonoral human conditions and di- ' seases .
  • composition and method can be used in the following, nonlimiting practical applications:
  • over-the-counter oral hygiene products e.g., mouth rinses and dentrifrices
  • oral hygiene products for the control of plaque and gingivitis using a combination of a berberine and an antibiotic
  • the combination of berberine chloride and an antibiotic, like CHX can be used in other oral products, including dental adhesives for orthodontic brackets to reduce plaque accumulation, dental impression material, chewing gum for caries and gingivitis control, press mints, and varnishes;
  • Berberine is a plant alkaloid having a long history of medicinal use in both Ayurvedic and Chinese medicine. Berberine is present in Hydrastis canadensis- (goldenseal) , Coptis chinensis (coptis or goldenthread) , Berberis aguifolium (Oregon grape) , Berberis vulgaris (barberry) , and Berberis aristata
  • Berberine can be found in the roots, rhizomes, and stem 1 bark of these plants. Berberine ' extracts and decoctions have demonstrated significant antimicrobial activity against a variety of organisms, including bacteria, viruses, fungi, protozoans, helminths, and chlamydia (Park et al . , 2001; Cowan, 1999; Grippa et al . , 1999; Park et al . , 1999) . The amount of berberine in OTC products is about 50 to 200 mg. Currently, the predominant clinical uses for berberine include treatment of bacterial diarrhea, intestinal parasite infections, and ocular trachoma infections (Harry et al .
  • Chlorhexidine is a bisbiguanide bio- cide, and compositions containing CHX have been used as topical disinfectants since the middle 1970s.
  • CHX targets the cytoplasmic membrane of bacteria resulting in the loss of structural organization and integrity, and ultimately the coagulation and precipitation of cytoplasmic constituents.
  • CHX has been used in ' postoperative rinsings, subgingival irrigation, and as an adjunct to mechanical debriment .
  • CHX has a low level of toxicity, both locally and systemically, and shows no permanent retention in the body (International
  • CHX usually is delivered in rinsing solutions (typically containing 1-2% CHX) , and slow-release devices such as varnishes and slab-like sustained delivery devices (Southard and Godowski , 1998) .
  • CHX also can be ' incorporated into chewing gum, toothpastes, and toothbrushes, or as a disinfectant for dentures in patients with candidal infections (International Health Care Foundation, 2001) .
  • Tetracyclines are broad-spectrum antibiotics that can be delivered by many routes of administration, including oral administration and sustained delivery systems. Tetracyclines inhibit protein synthesis by combining with the small (30S) subunit of the ribosome and inhibiting the binding of aminoscyl-tRNA molecules to the ribosomal A site (Prescott et al . , 1999). Tetracyclines generally are considered bacteriostatic . The number of re- ports directed to bacterial resistance to tetracyclines, both in vi tro and in vivo, is increasing, however, tetracyclines still are very useful and are routinely used in periodontal treatment. However, if resistant bacteria constitute potential pathogens, the periodontal disease may not be treated, but rather aggravated further (Slots and Rams, 1990) .
  • Tetracyclines concentrate in periodontal pockets at inhibitory levels 2-4 times higher than in the blood. Tetracyclines also strongly bind to root surfaces and can be released in active form over extended time periods. Sublethal concentrations of tetracyclines reduce adherence and coaggre- gation properties of a number of disease-associated bacteria, 'and also inhibit the collagenolytic enzymes of mammalian cells enabling periodontal ligament regeneration (Slots and Rams, 1990) .
  • the present invention shows: a) the antimicrobial activity of goldenseal alkaloids against representative oral pathogens and human pathogens, and b) the synergistic efficacy of berberine administration combined with administration of an antibiotic, like CHX, tetracycline (Tet) , colistin sulfate (Col) , or doxycycline (Dox) , against oral and other human pathogens .
  • an antibiotic like CHX, tetracycline (Tet) , colistin sulfate (Col) , or doxycycline (Dox)
  • the alkaloids extracted from goldenseal were berberine, hydrastine, schemaline, canadine and isocorypalmine .
  • a crude extract also was obtained.
  • a berberine derivative, i.e., berberrubine also was tested, but was not -isolated from goldenseal.
  • Berberine and hydrastine were formulated as a chloride salt and a hydrochloride salt, respectively, which eliminated solubility problems associated with the original compounds .
  • test organisms Five representative oral pathogens were used as the test organisms, i.e., Streptococcus mutans, Streptococcus gordonii , Porphymonas gingi - valis , Fusobacterium nucleatum, and Actinobacillus actinomycete comi tans . Cultures were obtained from . the University of Illinois-Chicago culture collection at the College of Dentistry. S . mutans, S . gordonii , and A . actinomycetemcomi tans were grown in Brain Heart Infusion broth (Difco, Becton Dickinson and Co., Sparks, MD, USA) . P.
  • gingivalis was grown using defined media containing (1 _1 ) :30 g Tryptic Soy Broth (Difco, USA) ; 5 g Yeast Extract (Difco, USA) ,- 0.5 g 1-cysteine hydrochloride (Sigma, USA); 0.3 ⁇ g menadione (Sigma, USA) and 5 ⁇ g hemin (Sigma, USA) .
  • F. nucleatum was grown in Schaedler Broth (Difco, USA) .
  • Three human pathogens also were tested, Escherichia coli , Staphylococcus aureus , and Pseu- domonas aeruginosa .
  • the antimicrobial activity of the alkaloids were evaluated using a microdilution assay in 96- well microtiter plates (Fisher Scientific, USA) .
  • the tested • compounds were berberine (extracted from goldenseal, B-G) , berberine chloride (commercial, Sigma, USA, B-S) , berberine chloride goldenseal (B- CG) , schemaline, hydrastine, hydrastine chloride, canadine, isocorypalmine, berberrubine (against S . mutans, A . actinomycetemcomi tans, and P.
  • gordonii were incubated aerobically (also E. coli , S . aureus, and P. aeruginosa) .
  • F. nucleatum and P. gingivalus were incubated anaerobically (anaerobic chamber, Ionia Scientific Inc., Marietta, OH) in 10% H 2 , 5% C0 2 , and 85% N 2 .
  • A. actinomycetemcomi tans was incubated both aerobically and anaerobically. Growth was monitored spectrophoto- metrically (A 660nm ) at 0, 24, and 48 h using a Power Wave 200 Microplate Scanning Spectrophotometer (BioTek Instruments, USA) .
  • the readings were captured and analyzed using the associated KC4 software, Version 2, Revision 12 (BioTek Instruments, USA) . Because berberine and other compounds, for example, berberrubine, are colored, the MIC of the test compounds was defined as the minimum concentration of compound limiting turbidity equal to the compound color blank.
  • the compound controls included inoculated growth medium without test compounds .
  • the MBC of the compounds against each test species was performed and determined as described in the previous section for MIC determination. After 48 h, 100 ⁇ l of culture was removed from wells containing the compound concentrations at or higher than the MIC. Serial dilutions (10 "1 to 10 "5 ) were performed, for each sample, and 100 ⁇ l of the appropriate dilutions was plated out in triplicate onto appropriate agar. After 48 h, and incubation at 37 °C under appropriate atmospheric conditions, the number of CFU ml "1 were enumerated. The MBC was defined as the lowest concentration of compound that killed 99.9 % of the test bacterial population. Combination determination
  • the alkaloids were tested in pair-wise combinations (fractional inhibitory concentrations, FIC) against all the bacteria to determine whether interactions were antagonistic (FIC>1) , additive (0.5 ⁇ F1C ⁇ 1) , or synergistic (FIC ⁇ 0.5).
  • FIC fractional inhibitory concentrations
  • the FIC was calculated by the following: (MIC A corab i nat i on /MIC aic ⁇ e) + (MIC B combinati on/MIC B a ⁇ one ) . Where A represents compound 1 and B represents compound 2.
  • the antibacterial efficacy of the alkaloids also were tested in combination with tetracycline HC1 (Tet- HC1) (0.2 to 25 ⁇ g ml "1 ) (Sigma Chemicals, St.
  • S . mutans Two representative organisms, S . mutans and F. nucleatum, were used in this study. Viability of S . mutans to berberine (B-CG) 500 and 1000 ⁇ ⁇ ml "1 ) ; CHX (20 ⁇ g ml “1 ) ; B (500 ⁇ g ml “1 ) with CHX (10 ⁇ g ml "1 ) ; and B (500 ⁇ g ml "1 ) with CHX (5 ⁇ g ml "1 ) was tested. Viability of F.
  • B-CG berberine
  • nuclea tum to B 500 and 1000 ⁇ g ml "1 ) ; Dox HCL (5 ⁇ g ml “1 ) ; B (500 ⁇ g ml “1 ) with Dox (2.5 ⁇ g ml rl ) ; and B (250 ⁇ g ml “1 ) with Dox (2.5 ⁇ g ml “1 ) also was tested.
  • the bacteria were prepared as previously described and the optical density was adjusted to give a ' concentration 1 x 10 CFU ml "1 . Aliquots of 4 ml of agent, prepared in sterile water, were placed into sterile test tubes and 1 ml of adjusted cells was added.
  • the control consisted of 4 ml of PBS and 1 ml of cells. Samples were removed at 0, 15, 30, and 60 minutes, serial dilutions were performed, and the appropriate dilutions plated out. The plates were incubated at 37°C under appropriate atmospheric conditions for up to 72 hours. Viable colony counts were determined and compared to nontreated control Percentage kill was calculated using the' formula:
  • Relative cell surface hydrophobicity of the test bacteria was evaluated using a modified method based on Rosenberg et al . (1980) .
  • the cells were collected by centrifugation (2400 x g, 10 minutes) , washed twice with phosphate buffered saline (PBS) (0.05 M, pH 6.8), adjusted to give a concentration of 1 x 10 s CFU ml "1 , and mixed with equal volumes of the test agents (Table 1) .
  • 500 ⁇ l of hexanes Sigma Chemicals
  • the reaction mixture vigorously agitated for 1 minute.
  • the optical density (O.D. A 550 nm) of the aqueous phase was determined.
  • the relative cell surface hydrophobicity was expressed as a percentage of the absorbance of the organic phase
  • the efflux pump inhibitor carbonyl cyanide m-chlorophenylhydrazone was added to the reaction mixture (final concentration 100 ⁇ M) .
  • the samples were immediately diluted with 1 ml ice-cold sodium phosphate buffer and then were centrifuged for 5 minutes at 5600 x g. The pellet was washed once with'l ml ice-cold buffer and resuspended in 1 ml of 0.1 M glycine hydrochloride (pH 3) for at least 15 hours at room temperature.
  • the fluorescence of the supernatant was measured (excitation 355 nm. and emission 517 nm) with a spectrofluori- meter (Model 810/814, Photon Technology International, Monmouth Junction, NJ) .
  • Hydrastine the major component (2-6% w/w) of goldenseal, did not inhibit growth of the test bacteria at concentrations ⁇ 500 ⁇ g ml "1 .
  • Canadaline, hydrastine, canadine, and isocorypalmine did not exhibit antimicrobial properties at concentrations ⁇ 250 ⁇ g ml -1 .
  • a . actinomycetemcomi tans was resistant to the crude extract and all the test compounds at concentrations ⁇ 500 ⁇ g ml "1 .
  • Berberrubine an analog of berberine, did not inhibit the growth of A . actinomycetemcomi tans at concentrations ⁇ 300 ⁇ g ml "1 .
  • berberrubine may be antimicrobial against S. mutans- at concentrations higher than 300 ⁇ g ml "1 because at lower concentrations growth was limited but not inhibited.
  • Berberrubine demonstrated antimicrobial activity against P. gingivalis at concentrations comparable to berberine (MIC 17 ⁇ g ml "1 ) .
  • This assay system allows concomitant evaluation of the minimum concentration that inhibits biofilm growth.
  • The' MIC values obtained for each species also represent the concentration that inhibited biofilm formation. There was no difference in antimicrobial activity between B-G and the other berberine compounds (B-S and B-CG) .
  • MIC values represent the mean of at least three separate . replicates
  • CHX was diluted from left to right at 10 to 0.08 ⁇ g ml "1 (rows 1 to 8), berberine was diluted, from top to bottom at 250 to 0.09 ⁇ g ml "1 (columns 1 to 8) .
  • Column 9 was the cell control; column 10 was berberine alone (250 to 0.09 ⁇ g ml "1 ); and column 11 was CHX alone (10 to 0.08 ⁇ g, ml "1 ) .
  • Row H was the solvent control (8 % DMSO, HI to H8) .
  • Column 12 was the media blank.
  • the dilution assay was designed such that Al contained the highest concentration of both agents (berberine 250 ⁇ g ml "1 and CHX 10 ⁇ g ml "1 ) .
  • the final concentration of berberine in wells A2 to A8 was 125 ⁇ g ml "1 .
  • the FIC value was calculated from well C6. FIC was calculated by: (MIC
  • SD indicates a signif icant difference in the reduction of viability between treatments at a 95% confidence interval .
  • NSD indicates no significant difference in the reduction of viability between treatments at a 95% confidence interval .
  • SD indicates a significant difference in the reduction of viability between treatments at a 95% confidence interval .
  • NSD indicates no significant difference in the reduction of viability between treatments at a 95% conf idence interval .
  • the assay demonstrated that A . actino- bacillus was less hydrophobic compared to the other bacteria. When A . actinobacillus was exposed to the agents in combination and alone, an increase surface hydrophobicity of about 25% was observed.
  • Plants remain a major source of medicinal compounds. It is estimated that 20,000 plant species are used for medicinal purposes (Penos, 1983). Medicinal plants have been used throughout history, and efficacy often is associated with anecdotal folklore. Further estimations suggest that 74% of 119 plant derived drugs were discovered as a result of chemical studies to isolate the active components responsible ' for their traditional use (Farnsworth et al., 1985) . At least 25% of recent medicinal prescriptions contain one active compound from plant species (Haq, 2000) , and the estimated value of these prescriptions containing plant-derived compounds is worth about $10. billion dollars in the U.S. alone (Duke, 1990).
  • the present invention highlights ' the importance of identifying the major antimicrobial contributions of alkaloids from plants, and illustrates that use of extract components in combination with standard antibiotics provides a synergistic therapeutic effect .
  • Biofilm formation takes place when bacteria attach to a surface from a liquid phase, producing exopolymeric substances that aid in the continued attachment to the surface and offer protection from the external environment.
  • Bacteria in the form of biofilms have been reported to be up to 1000 times more resistant to treatment than the bacteria present in liquid phase (Lewis, 2001; Costerton et al . , 1999). This is a serious problem for the efficacious treatment of infections (Costerton et al . , 1999).
  • Harry et al . , (1988) reported that berberine sulfate at sub-MIC levels inhibited the adherence of S . pyogenes to epithelial cells. The results of these tests have shown that berberine and combinations thereof were effective at inhibiting biofilm formation o'f all the test species except A . actinomycetemcomi tans .
  • a . actinomycetemcomi tans, a facultative anaerobe and potent periodontal. pathogen is known for its resistance to antimicrobial treatment, particularly tetracycline antibiotics (Fives-Taylor et al . , 2000) . Reversal of resistance by synergy produced as a result of the combination of conventional antibiotics and nonantibiotics has been achieved (Kristiansen et al . , 1997). Although the results from this investigation have shown that A . actinomycetemcomi tans was not susceptible to this antimicrobial regime, it may be possible to achieve an antimicrobial response if the concentration of agents was increased.
  • P. aeruginosa increasingly is associated with nosocomial infections both in the U.S. and Europe, particularly pneumonia, and can be fatal for immunoco promised persons.
  • Pseudomonas infections can be spread within hospitals by healthcare workers, medical equipment, sinks, disinfectant solutions, and food.
  • the bacterium also causes serious problems in individuals suffering from cystic fibro- sis,. a fatal genetic disorder. .
  • Colistin is a cationic antibiotic belonging to the polymyxin group. Colistin initially was introduced in the early 1960s, but became redundant by the early 1970s due to reports of its severe toxicity. In this present study, this strain of P. aeruginosa was resistant to berberine at the test concentrations, although impairment of growth was observed when combined with CHX and Col . As with the other antibiotics, Tet and Dox, adverse side effects associated with Col can be reduced when combined with berberine. Other strains of P. aeruginosa may demonstrate greater susceptibility and, therefore, the beneficial effects of combining Col with berberine cannot be disregarded.
  • CHX ' is a widely used biocide in antiseptic and disinfectant products', particularly in hospitals. It has broad spectrum efficacy and is much less irritating to tissue than other products (McDonnell et al . , 1999). Chlorhexidine is bactericidal and fungicidal to a wide variety of organisms and it is possible that by combining CHX with berberine, that its activity spectrum may be increased. Bacterial resistance to CHX has been reported, specifically S . aureus and S . epidermidis, and there is increasing concern regarding the link between biocide exposure and antibiotic resistance (Russell et al . , 2000; Russell, 2000; McDonnell et al.,, 1999; Russell et al . , 1996). Combinations of CHX with a natural antimicrobial agent, such as berberine, may result in a more environmentally acceptable product and may serve to increase the longevity of use and efficacious treatment of oral infections and disinfectant procedures .
  • a natural antimicrobial agent such as
  • MDR pump systems that can export structurally unrelated antibiotics (Sulavik et al . , 2001) . MDR systems have also been identified in S . aureus (Markham et al . , 1999), P. aeruginosa (Li et al . , 1995), and A . actinomycetemcomi tans (Fives- Taylor et al., 2000).
  • MDR pump systems are usually ATP dependent.
  • the increased susceptibility of the anaerobic species to berberine and combinations thereof may be related to the reduced amounts of ATP produced under anaerobic conditions (Prescot et al . , 1999) .
  • a goldenseal alkaloid can produce toxic symptoms ranging from mouth irritation, nausea, vomiting, and diarrhea to cardiac depression and ⁇ central nervous system paralysis (Tice, 1997) . It. also has been reported that berberine (100 mg) reduced the efficacy of tetracycline in people with cholera by reducing tetracycline absorption (Khin- Maung et al . , 1985). However, in another report the findings were inconclusive • as to whether berberine significantly interacted tetracycline antibiotics (Raddani et al . , 1987).
  • CHX ' is the "gold standard" of all plaque control agents and is included in oral hygiene products.
  • CHX is a prescription drug in the US.
  • CHX has many side effects, including undesirable . taste, tooth discoloration, and increased calculus (tartar) formation in users, although no permanent retention in the body is observed (International Health Care Foundation, 2001) .
  • Tetracyclines are conventional antibiotics and are routinely used in the treatment of periodontal disease because of their unique , ability to accumulate in the pocket area. However, like many conventional antibiotics, many bacteria, especially important human pathogens, have developed • a resistance to tetracyclines. Tetracyclines also have many side effects including, gastrointestinal discomfort, nausea, vomiting, diarrhea, and permanent tooth discoloration in children up to 8 years old.
  • Berberine is found in many health supplements in amounts exceeding 50 mg.
  • Antibiotics used in periodontal treatment include tetracycline and doxycycline, and are administered typically at 100 mg (doxycycline) to 500 mg (tetracycline) with concentrations in the gingival crevicular fluid ranging from 1 to 8 ⁇ g ml "1 (doxycycline) and 2 to 12 ⁇ g ml "1 (tetracycline) (Slots and Rams, 1990) .
  • the present invention shows that when berberine is administered in combination with an antibiotic, like CHX, tetracycline, or doxycycline, approximately half to one quarter the amount of a berberine and an antibiotic is required to achieve the desired growth inhibition compared to when either agent is used alone. Synergy observed in vi tro also is evident in vivo, and can lead to the development of drug combinations for the management of infections that are difficult to treat with one antibiotic alone (Kristiansen and Amaral, 1997). If a combination treatment using berberine and a conventional antimicrobial agent is employed, decreased concentrations of both agents can be used, thereby reducing the occurrence and/or severity of adverse side effects.
  • an antibiotic like CHX, tetracycline, or doxycycline
  • Dental plaque frequently is associated with oral diseases including dental caries and periodontal (gum) disease. Plaque control is of utmost importance for the prevention of these diseases . Plaque control can be achieved by mechanical (e.g., brushing. or flossing) and chemical means. For the latter, antimicrobial agents have been incorporated into oral hygiene products, such as toothpastes and mouth rinses, to control plaque and gingivitis/- periodontal disease. Therefore, a composition containing reduced levels of CHX, tetracycline, doxycycline, or other antibiotics, that minimizes occurrence or severity of the above-mentioned ad- verse effects while achieving the desired antimicro- ⁇ bial effect would be a significant advance in the art .

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Abstract

L'invention concerne une composition et un procédé pour réguler des pathogènes oraux ou tout autre pathogène humain. La composition et le procédé de l'invention comprennent l'utilisation d'un antimicrobien ou d'un antibiotique et de la berbérine en tant qu'agents actifs pour traiter des mammifères, en particulier des êtres humains.
PCT/US2004/008616 2003-04-03 2004-03-22 Procede et composition pour reguler des pathogenes oraux WO2004093876A2 (fr)

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Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20090047620A1 (en) * 2005-11-17 2009-02-19 Thomas Klettke Anti-microbial dental impression material
CN102079765A (zh) * 2010-12-15 2011-06-01 西南大学 9-o-糖苷-小檗碱盐及其制备方法和应用
US20120165357A1 (en) * 2009-06-30 2012-06-28 Derman Biomedicine Co. Ltd. Compositions Containing Berberine or Analogs Thereof for Treating Rosacea or Red Face Related Skin Disorders
CN105497028A (zh) * 2015-12-15 2016-04-20 上海壹志医药科技有限公司 小檗红碱的药物用途
WO2017143361A1 (fr) * 2016-02-18 2017-08-24 Drabinsky Michael Earl Traitement de maladies causées par les spirochètes
WO2018176079A1 (fr) * 2017-03-28 2018-10-04 Iriccorgpharm Pty Ltd Alcaloïdes berbérine dans la prévention et/ou le traitement d'une maladie intestinale
WO2019196957A1 (fr) * 2018-04-13 2019-10-17 中国医学科学院药物研究所 Dérivé de type berbérine hydrophile et son application dans la préparation d'un médicament
RU2812224C1 (ru) * 2023-03-09 2024-01-25 Федеральное государственное бюджетное образовательное учреждение высшего образования "Ставропольский государственный медицинский университет" Министерства здравоохранения Российской Федерации (ФГБОУ ВО СтГМУ Минздрава России) Противомикробное аппликационное средство для лечения воспалительных заболеваний пародонта с контролируемым высвобождением

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US5741138A (en) * 1996-08-12 1998-04-21 The Procter & Gamble Company Oral compositions
WO1998044926A1 (fr) * 1997-04-04 1998-10-15 Optiva Corp. Compositions antimicrobiennes
US6248309B1 (en) * 1997-04-04 2001-06-19 Optiva Corporation Gums containing antimicrobial agents
WO2002091848A1 (fr) * 2001-05-15 2002-11-21 The Procter & Gamble Company Compositions de confiserie

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US5741138A (en) * 1996-08-12 1998-04-21 The Procter & Gamble Company Oral compositions
WO1998044926A1 (fr) * 1997-04-04 1998-10-15 Optiva Corp. Compositions antimicrobiennes
US6248309B1 (en) * 1997-04-04 2001-06-19 Optiva Corporation Gums containing antimicrobial agents
WO2002091848A1 (fr) * 2001-05-15 2002-11-21 The Procter & Gamble Company Compositions de confiserie

Cited By (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20090047620A1 (en) * 2005-11-17 2009-02-19 Thomas Klettke Anti-microbial dental impression material
US8933147B2 (en) * 2005-11-17 2015-01-13 3M Innovative Properties Company Anti-microbial dental impression material
US20120165357A1 (en) * 2009-06-30 2012-06-28 Derman Biomedicine Co. Ltd. Compositions Containing Berberine or Analogs Thereof for Treating Rosacea or Red Face Related Skin Disorders
US9486402B2 (en) * 2009-06-30 2016-11-08 Derman Biomedicine Co. Ltd Compositions containing berberine or analogs thereof for treating rosacea or red face related skin disorders
CN102079765A (zh) * 2010-12-15 2011-06-01 西南大学 9-o-糖苷-小檗碱盐及其制备方法和应用
CN105497028A (zh) * 2015-12-15 2016-04-20 上海壹志医药科技有限公司 小檗红碱的药物用途
WO2017143361A1 (fr) * 2016-02-18 2017-08-24 Drabinsky Michael Earl Traitement de maladies causées par les spirochètes
WO2018176079A1 (fr) * 2017-03-28 2018-10-04 Iriccorgpharm Pty Ltd Alcaloïdes berbérine dans la prévention et/ou le traitement d'une maladie intestinale
WO2019196957A1 (fr) * 2018-04-13 2019-10-17 中国医学科学院药物研究所 Dérivé de type berbérine hydrophile et son application dans la préparation d'un médicament
US11725006B2 (en) 2018-04-13 2023-08-15 Institute Of Materia Medic, Chinese Academy Of Medical Sciences Hydrophilic berberine-type derivative and application thereof in preparing drug
RU2812224C1 (ru) * 2023-03-09 2024-01-25 Федеральное государственное бюджетное образовательное учреждение высшего образования "Ставропольский государственный медицинский университет" Министерства здравоохранения Российской Федерации (ФГБОУ ВО СтГМУ Минздрава России) Противомикробное аппликационное средство для лечения воспалительных заболеваний пародонта с контролируемым высвобождением

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