WO1984003624A1

WO1984003624A1 - Method for inhibiting the growth of anaerobes

Info

Publication number: WO1984003624A1
Application number: PCT/US1984/000415
Authority: WO
Inventors: Samuel L Yankell; Catherine C Schifter; Irving M Shapiro
Original assignee: University Patents Inc
Priority date: 1983-03-17
Filing date: 1984-03-16
Publication date: 1984-09-27
Also published as: EP0138969A1

Abstract

A method for inhibiting the growth of anaerobes which comprises contacting said anaerobes with an oxygenated composition comprising one or more polyfluorinated organic compounds.

Description

METHOD FOR INHIBITING THE GROWTH OF ANAEROBES

This application is a continuation-in-part of our copending Application Serial No. 476,438, filed March 17, 1983.

This invention relates to methods for inhibiting the growth of anaerobes, and more particularly to methods for inhibiting the growth of anaerobes with oxygenated polyfluorinated organic compounds.

All organisms may be classified into one of four traditional groups: 1)obligate (strict) aerobes, which require the presence of free oxygen to grow; 2)obligate (strict) anaerobes, whose growth is severely or totally restricted by free oxygen; 3)facultative anaerobes, which can grow with or without free oxygen, and 4)microaerophiles, which require the presence of small amounts of free oxygen to grow but which are inhibited by larger amounts of free oxygen. It has been noted that many obligate aerobes and facultative anaerobes are inhibited when cultured in the presence of pure 0₂ rather than air; the reason for this 0₂ toxicity is not known. For the purposes of this invention, the unmodified term "anaerobe" means any eucaryotic or procaryotic cell whose growth can be inhibited by the presence of free oxygen, including but not limited to all cells traditionally classified as obligate anaerobes and microaerophiles, and those obligate aerobes and facultative anaerobes which are inhibited by pure 0₉. Such anaerobes are found in environments with low oxygen tension, such as in the muds and sediments of lakes, rivers, and oceans; in water-logged soils; in sewage; deep underground in oil pockets and sources of rivers and springs; in canned foods; and in the oral cavity and intestinal tracts of insects and animals. Anaerobes may also be found in lo¬ calized pockets of low oxygen tension within larger aerobic environments, e.g. within deep narrow wounds or within necrotic tissue. Many anaerobes are patho¬ genic or produce extremely toxic metabolites.

Where the presence of anaerobes or their meta¬ bolites is undesirable, either the anaerobes or the anaerobic environment must be destroyed. This is often accomplished by massive aeration — the intro¬ duction into the anaerobic environment of large volumes of air or oxygen accompanied by stirring or mixing to achieve uniform dispersion of the gas. Anaerobes can also be killed by traditional steri- lants such as heat, radiation, or toxic chemicals. However, when the environment is not suited to massive aeration, or too delicate for the application of traditional sterilants, the method of choice has been the application of antibiotics. Unfortunately, such treatment frequently kills other, desirable, organisms in the environment as well. Thus, there is a need for a method of selectively inhibiting the growth of anaerobes within a particular environment.

It has now been discovered that the growth of anaerobes can be selectively and efficiently inhibit¬ ed by contacting said anaerobes with an oxygenated composition comprising one or more polyfluorinated organic compounds. Compositions comprising one or more polyfluorinated organic compounds will be referred to as "polyfluorinated compositions".

By "polyfluorinated organic compound" is meant any organic compound containing two or more fluorine radicals. Organic compounds in which 50% or more of the possible substitution sites on the molecule are occupied by fluorine are preferred. Perfluoro- co pounds are particularly preferred. The term "perfluoro" when used in conjunction with the name of a compound indicates that all hydrogens in that compound have been replaced by fluorine. Poly¬ fluorinated compounds are characterized by low boiling points, low surface tensions, high densities, low refractive indices, low dielectric constants, and low sound velocities. They are insoluble in water and alcohols, and soluble in aliphatic hydrocarbons, ethers, and chlorinated solvents. These compounds have a high oxygen-carrying capacity, low toxicity, chemical and thermal stability, and are eliminated from the body without being extensively metabolized (i.e. , they are relatively metabolically inert) . In addition, these compounds readily release dissolved oxygen to surrounding tissues in vivo. These prop¬ erties are accentuated as the degree of fluorination increases. Polyfluorinated organic compounds and their methods of 'manufacture are known, and, per se, form no part of the present invention. See, e.g. , Hudlicky, M. , Chemistry of Organic Fluorine Compounds (MacMillan Co., New York, 1962); Lovelace, A.M., D.A. Rausch, and W. Postelnek, Aliphatic Fluorine Com¬ pounds (Reinhold Pub. Corp., New York, 1958); Sheppard, W.A. , and CM. Sharts, Organic Fluorine Chemistry (W.A. Benjamin, Inc., New York, 1969); Simons, J.H. (ed.). Fluorine Chemistry, Vols. I, II, V (Academic Press, New York, 1950); Stacey, M. , J.C. Tatlow, and A.G. Sharpe (eds.). Advances in Fluorine Chemistry, Vols. I-V, (Butterworths, London); Tarrant, P. (ed.). Fluorine Chemistry Reviews, Vol. I (Marcel Dekker, New York, 1967) , for examples of compounds suitable for use in this invention, and methods of making same.

Among the polyfluorinated organic compounds which are preferred are polyfluorinated compounds such as:

perfluorodecalin of the formula:

perfluorotrimethylbicyclo[3.3.1]nonane of the formu¬ la:

perfluoro-N,N-dimethylcyclohexylmethylamine of the formula :

perfluoro-2,2,4,4-tetramethylpentane of the formula:

CF₃-C(CF₃)₂-CF₂-C(CF₃)2" ^F ₃

perfluoro-N-methyldecahydroquinoline of the formula:

perfluoro-1-methyloctahydroquinolizine of the formu¬ la:

perfluoro-N-cyclohexylpyrrolidine of the formula:

F₂

perfluoro-1,3,5,7-tetramethyladamantane of the formula:

l,2-di(perfluoro-n-butyl)ethylene of the formula:

CF₃-(CF₂)₃-CH=CH-(CF₂)₃-CF^ , perf luorobutyltetrahydrofurane of the formula:

perfluoro-1-bromo-n-octane of the formula:

CF₃- (CF₂ ) ₆-CF₂Br

perfluorotri(rι-butyl)a ine of the formula:

N(CF₂-CF₂-CF₂-CF₃)₃

perfluorotri(rι-propyl)amine of the formula:

N(CF₂-CF₂-CF₃)₃

perfluoro-1,4-dibromo-rι-butane of the formula:

BrCF₂-(CF₂)₂-CF₂Br ,

perfluoro-1,2-dibr^'omoethane of the formula:

BrCF₂-CF_?Br

perfluoro-1-bromocyclohexane of the formula:

perfluoro-l,2-dibromocyclohexane of the formula:

perfϊuoro-l,2-dibromopropane of the formula:

CF₃-CFBr-CF₂Br

perfluoro-1-bromo-rι-hexane of the formula:

CF₃-(CF₂)₄-CF₂Br

dichlorodifluoromethane of the formula:

CC1₂F₂

chlorotrifluoromethane of the formula:

CC1F-

chlorodifluoromethane of the formula:

CHCIF-

trifluoromethane of the formula:

CHF-

1,1-difluorotetrachloroethane of the formula:

CC1F₂-CC1₃ , 1 ,2-dif luorotetrachloroethane of the formula :

CCl-F-CCl-F

1,1,1-trichlorotrifluoroethane of the formula:

CC1₃-CF₃

1,2-dichlorotetrafluoroethane of the formula:

CC1F₂-CC1F₂

perfluoro-1-chloroethane of the formula:

CC1F₂-CF₃

perfluorocyclobutane of the formula:

dibromodifluoromethane of the formula:

CBr₂F₂

bromotrifluoromethane of the formula:

CBrF-_j perfluorocyclohexane of the formula:

F.

perfluoro(methylcyclohexane) of the formula:

F.

perfluorotoluene of the formula:

In addition, polyfluorinated compositions comprising.

Fluosol-DA (TM) [Alpha Therapeutic Corp., Los Angeles,

CA] or members of the 3M "FC" series are also suit¬ able for use in this invention. Fluosol-DA is an aqueous emulsion comprising 14% (w/v) perfluoro- decalin, 6% (w/v) perfluorotripropylamine, an emulsifier, and small amounts of inorganic salts and other ingredients. The 3M "FC" series is a class of compositions, said compositions each consisting of one or more pure perfluorocompounds (e.g. FC-72, FC-84, FC-77, FC-104, FC-75, FC-40, FC-43, FC-70, FC-71, FC-80, FC-47, all trademarks of, and available from 3M Co., St. Paul, MN) .

The polyfluorinated compounds or compositions may be used individually or in combination with one or more other polyfluorinated compounds or composi¬ tions. The polyfluorinated organic compound(s) may be used in pure form, i.e. in a composition compris¬ ing about 99-100% polyfluorinated organic com¬ pound(s) , or as the active agent(s) in a mixture, suspension, solution, or emulsion in combination with a solvent or carrier. The polyfluorinated composi¬ tions may also contain minor amounts of other ingredients such as sequestering agents, e.g. up to 1% (w/v) EDTA or up to 1% (w/v) citric acid; thickeners, e.g. hydroxyethyl starch (McGaw Labo¬ ratories, Glendale, CA 91201); and/or one or more emulsifying agents, e.g. Pluronic F-68 (TM) [a conden¬ sate of ethylene oxide with a hydrophobic base formed by condensing propylene oxide with propylene glycol, Wyandotte Chemical Corp., Wyandotte, MI 48192]. In order to be effective, these polyfluorinated compositions must be oxygenated, i.e., free 0₂ must be dissolved in the compositions. This can be accomplished by aeration with air or oxygen, pref¬ erably under pressure, just prior to use. Alter¬ natively, the compositions can be packaged in indi¬ vidual vials under oxygen pressure, and the vial can be shaken thoroughly to disperse and dissolve the oxygen in the polyfluorinated composition immediately prior to use. The maximum oxygen tension possible is determined primarily by the temperature and the polyfluorinated organic compound(s) selected. Methods of measuring the concentration of oxygen dissolved in the polyfluorinated composition and of calculating the maximum oxygen tension are known see, e.g. , Osburn, J. O. , Fed. Proc. 29(5) 1704-1707 (1970); Reed, T.M. , III, Fed. Proc. 29(5) 1708-1713(1970); Peterson, R. E. , Fed. Proc. 29(5) 1714-1716(1970); "3M Company: Determination of the Solubilities of Gases in Perfluorocarbon Fluids," Fed. Proc. 29(5): 1817-1818(1970); Mears, W.H., and R. L. Beavers, Fed. Proc. 29(5): 1819(1970), all incorporated herein by reference.

In addition to being employed in pure form, or as a mixture, solution, suspension, or emulsion, the polyfluorinated compositions of the present invention can also be employed in the form of an aerosol. In this embodiment, the composition is confined within a container, under oxygen pressure, with one or more suitable aerosol propellants. ^* Suitable propellants are well known in the aerosol formulation art and include, for example, fluorinated hydrocarbons, e.g. , trichlorofluoromethane (Freon 11 ') and dichloro- fluoromethane (Freon 21 (*TMΪ'). Alternatively, the polyfluorinated organic compounds dichlorodifluoro- methane (e.g. Freon 12 '), chlorotrifluoro ethane (e.g. Freon 13 (^vTM)'), chlorodifluoromethane (e.g.

Freon 22^(TM)), trifluoromethane (e.g. Freon 23^(TM)), trichlorotrifluoroethane (e.g. Freon 113 (TM)'), dichlorotetrafluoroethane (e.g. Freon 114 (TM)'), chloropentafluoroethane (e.g. Freon 115 (TM) ) , per- fluorocyclobutane (e.g. Freon C318 (TM)), dibromodi- fluoromethane (e.g. Freon 12B2 (¹TM)'), and bromotri- fluoromethane (e.g. Freon 13B1 (TM) ) , are all them¬ selves suitable for use as propellants and can be formulated to function as both active compounds and propellant in the perfluorinated composition. The Freon compounds are a trademark of, and are available from, E.I. duPont deNemours Co., Wilmington, DE.

The polyfluorinated composition (i.e. the pure active agent(s) , mixture, solution, suspension, emulsion, or aerosol) is formulated to provide from about 5 to about 99% (w/v) polyfluorinated organic compounds, preferably from about 5 to about 25% (w/v) .

According to the method of the present inven¬ tion, the growth of anaerobes is inhibited by con¬ tacting said anaerobes with an oxygenated polyfluo¬ rinated composition formulated as above. The degree and rate of inhibition are primarily determined by the specific target anaerobe and the concentration of oxygen in the polyfluorinated composition; the higher the . oxygen concentration, the more rapid and effi¬ cient the inhibition. Under appropriate conditions, inhibition can be total, i.e. the anaerobes will die. Methods of determining the appropriate oxygen concen¬ tration to be used to inhibit specific anaerobes are known to those skilled in the art. Where the anaerobes are present in or on a living body, the oxygenated polyfluorinated compositions are administered in medical/dental applications according to procedures known in the art and used heretofore in administering antiseptic solutions. Typically, the affected tissues are perfused for several minutes with an amount of oxygenated polyfluorinated composition sufficient to flush the entire affected area. Preferably, the polyfluorinated organic compound(s) used are non-toxic to said body.

A further understanding of this invention can be had from the following non-limiting Examples. Unless expressly stated to the contrary, all operations were performed at ambient temperature (approximately 20

C) and pressure (approximately 1 atm) .

EXAMPLE 1

The representative polyfluorinated organic compound perfluorotributylamine can be purchased from, e.g. Alpha Therapeutic Corp., Los Angeles, CA M TM

(FC-43 ) or 3M Co., Minneapolis, MN (FC-47ⁱ ) . FC-43 is commercially supplied in a kit consisting of two solutions which must be mixed. The initial compositions of the solutions are as follows:

Solution A

perfluorotributylamine 25 g

Pluronic F-68 ^{(TM) ~} 3.2 g

Pyrogen-free distilled water....to 100 ml.

[Wyandotte Chemical Corporation, Wyandotte, MI] Solution B

NaCl 0 .600 g

MgC 1₂ 0.020 g

CaCl₂ 0 .028 g

Glucose 0.180 g

Hydroxyethyl Starch 3. 0 g

Pyrogen-free distilled water. . . to 14 ml .

FC-43 is formed by the addition of 6 mis . of solution A to 14 mis . of Solution B , to give final concen¬ trations as follows :

FC-43 % (w/v) perfluorotributylamine 20.0

Pluronic F-68 2.56

NaCl 0.60

KC1 0.034

MgCl₂ 0.020

CaCl₂ 0.028

NaHC0₃ 0.21

Glucose 0.180

Hydroxyethyl Starch 3.0

EXAMPLE 2

The term "chronic inflammatory periodontal disease" (CIPD) refers to a number of clinical symptoms which affect the gums and supporting tissues of the teeth and which, if untreated, often lead to eventual tooth loss. The exact etiological factors of many of these clinical symptoms have not yet been identified, however, most of the available evidence indicates that dental microbial plaque is the primary factor associated with both the initiation and progress of CIPD. CIPD is progressive and begins with inflammation . of the gum line (gingivitis) , followed by mild, moderate, and finally severe periodontitis, characterized by the formation of periodontal pockets around the affected tooth, inflammation, loss of bone, and eventual loosening and loss of the tooth.

It is well established that there is a distinct difference between the microbial flora present in healthy gingival crevices and the flora present in crevices evidencing gingivitis or mild to advanced periodontitis. In the healthy mouth the majority of bacteria in the oral cavity and in the gingival crevices are aerobic. As CIPD progresses, the bacterial population in the periodontal crevices or pockets gradually becomes dominated by gram-negative and anaerobic microorganisms.

With the identification of anaerobic microflora as etiologic agents of CIPD, a number of treatments have been utilized to remove or destroy these bacte¬ ria. Mechanical debridement has been the tradition¬ ally accepted method of treating CIPD and has been demonstrated to be effective in temporarily reducing the microbial population and alleviating the clinical symptoms. However, this is a tedious and painful procedure, and individual variations in tooth and pocket morphology and operator skill make the com¬ plete elimination of subgingival microflora virtually impossible. As a result, the remaining bacteria, and particularly the anaerobes, re-establish themselves in the periodontal pockets and precipitate recurring infections. Antibiotics, with or without debridement, have also been used to control or eliminate CIPD. Sys¬ temic treatments allow the antibiotic to reach the subgingival flora from the bloodstream via the gingival crevicular fluid. Broad spectrum antibiot¬ ics are generally used unless the specific microbial population involved has been identified. The draw¬ back to this approach is that although the target microorganisms are concentrated in a relatively small area of the body, high circulating levels are re¬ quired to achieve an effective local concentration of antibiotic. As a result, the normal microflora of the alimentary canal and gastrointestinal tract are also affected. Disturbance of the delicate balance of the normal microbial populations leads to over¬ growth by certain organisms, causing gastrointestinal distress and/or secondary infections of the mouth and throat. Repeated antibiotic treatments can aggrevate this effect.

Another method of treatment which has been explored has been the use of topical antiseptics, e.g. chlorhexidine or peroxide. These agents are generally non-specific and therefore tend to have the same drawbacks as antibiotics. Peroxide has been shown to reduce the anaerobic population,decrease the gingival fluid flow, and decrease post-extraction bacteremia when introduced into deep periodontal pockets. Peroxide rinses are frequently supplemented with salts, sodium bicarbonate, and/or antibiotics. However, peroxide is a strong oxidizing agent and in addition to having an indiscriminate antiseptic effect tends to damage the oral tissues.

The negative effects of the long-term use of antibiotics and antiseptics for treatment of CIPD are

O PΪ m IPO , reviewed in Accepted Dental Therapeutics_r 39th Edition American Dental Association, 1982, particu¬ larly pp. 375-376.

It has now been discovered that perfusion of gingival crevices or periodontal pockets with oxygenated polyfluorinated organic compounds substan¬ tially reduces the population of anerobic microflora therein.

Subgingival human plaque was taken from each of 2 subjects selected from a group of patients at the University of Pennsylvania School of Dental Medicine. Subjects were selected based on the following crite¬ ria:

1. Clinical and radiographic diagnosis of CIPD;

2. No medical history of antibiotic therapy within the past 12 months;

3. No oral prophylaxis within the past 6 months.

The plaque from each patient was then divided into 6 samples and each set of 6 was divided into three groups of two. Each sample was suspended in 0.1 ml of sterile aqueous 0.85 N sodium chloride. Within 15 minutes of collection, each sample was dispersed by aspirating and expelling the suspension three times using a 1 ml hypodermic syringe fitted with a 25-gauge needle. One drop was placed on a slide and coverslipped. Each sample was then examined for the population of bacteria present, according to the method of Listgarten, M.A. , and L. Hellden, J. Clin. Perio. 5:115-132 (1978). One hundred bacteria were counted at random and grouped on a percentage basis according to their morphology as motile rods, spirochetes, and all others. This procedure was repeated twice more and the average percentages were recorded for each group.

The materials tested were unoxygenated FC-43 and oxygenated FC-43 (oxygenated with pure oxygen for 30 seconds immediately prior to use) . After the initial counts were performed, the agents were randomly assigned for testing and approximately 0.1 ml of the test or control material was perfused under the coverslip of each slide with a 1-ml hyperdermic syringe fitted with a 23-gauge needle. After five minutes, each sample was recounted.

The results are presented in Table I. The average counts of motile rods and spirochetes in each sample group prior to treatment was similar. After treatment with either water or unoxygenated FC-43 there were no major differences in percentage counts of either motile rods or spirochetes. By contrast, after treatment with the oxygenated FC-43 the percent of motile rods decreased by 74.7% and spirochetes decreased 46.6% compared to initial percentages of total counts of these two microbial species, demon¬ strating the efficacy of this representative oxygenated polyfluorinated composition in inhibiting the growth of anaerobes. The fact that the unoxygenated FC-43 shows no activity demonstrates that the activity of the oxygenated FC-43 is not due to the presence of the sequestering agent, the salts or the carbohydrates in the FC-43. As an additional control, Solution A and Solution B as defined in Example 1 were each tested separately, with and without oxygenation, in a manner similar to that above. Both of the unoxygenated solutions and the oxygenated Solution B were inac¬ tive,1^, did not produce a major difference in percentage counts of motile rods or spirochetes, while the oxygenated Solution A showed results similar to those of the oxygenated FC-43.

Table I

Effect of FC-43 on Motile Rods and Spirochetes in Plaque Samples from CIPD Patients

EXAMPLE 3

In a manner similar to Example 2, the effect of hydrogen peroxide was assessed on plaque samples taken from two individuals with CIPD. Only single control values from each patient were counted. The test agent (hydrogen peroxide) was tested on dupli¬ cates of each plaque sample.

The results are shown in Table II. Average initial percentage counts of spirochetes and motile rods were similar for both the test and control groups. After treatment with hydrogen peroxide, the motile rods and spirochetes showed decreases of 93.3% and 79.5% respectively.

Comparison of the results of Examples 2 and 3 shows that perfusion with oxygenated polyfluorinated organic compounds, as represented by FC-43, achieves 80% of the decrease in motile rods and 59% of the decrease in spirochetes obtained with use of the popularly employed hydrogen peroxide. This result is achieved, however, without the concomitant oxidation of tissue which is known to result from hydrogen peroxide irrigation.

Table II

Effect of Hydrogen Proxide on Motile Rods and Spirochetes in Plaque Samples from CIPD Patients

₂2 patients x 1 control each 2 patients x 2 test samples each

EXAMPLE 4

Since plaque samples from different individuals have been observed to vary in susceptibility to treatment with antibacterial agents, the effects of hydrogen peroxide and oxygenated FC-43 on identical plaque samples were compared. The assay was con¬ ducted in a manner similar to Example 2. A sample was taken from an individual with CIPD, and divided into two groups of duplicates. Since initial amounts of motile rods were low in this individual's samples, counts of this organism were not included in the assay. The FC-43 was oxygenated with air, under pressure, from an air hose connected to the air/water accessory system of a dental drill, for 30 seconds immediately prior to use.

The results are shown in Table III. Initial counts of spirochetes were similar for the test and control groups. After addition of hydrogen peroxide, a 53% decrease in spirochete count relative to initial values was obtained. Treatment with oxygen¬ ated FC-43 resulted in a relative decrease in spiro¬ chete count of 50%. Again, these results show oxygenated polyfluorinated organic compounds to be as effective as hydrogen peroxide in reducing anaerobic bacterial populations, and these results are achieved without the potential negative side effects of peroxide treatment. Table III

Effects of Oxygenated FC-43 and Hydrogen Peroxide on Spirochetes in Plaque Samples from CIPD Patients

The oxygenated polyfluorinated compositions of the present invention are administered in med¬ ical/dental applications according to procedures known in the art and used heretofore in administering antiseptic agents. Typically, the infected periodontal tissues are perfused for several minutes with about 0.1 to about 1.0 ml. of the oxygenated polyfluorinated composition. This is accomplished by irrigation of the gingival crevice or periodontal pocket by means of a syringe or specially designed irrigation instrument, e.g., an Imax Irrigator/Medicator (TM) (Dental Scientific Systems,

Reston, VA) . The oxygenated polyfluorinated composi¬ tion may be administered either by the patient in the home, preferably in conjunction with recognized recommended oral hygiene practices such as tooth brushing and flossing, or by the dental clinician in the office as part of routine dental therapy.

As is demonstrated by these Examples, anaerobes are effectively inhibited by contact with composi¬ tions comprising one or more oxygenated polyfluorinat¬ ed organic compounds. As a particular example, the anaerobic motile rods and spirochetes which are associated with CIPD are inhibited by contact with such oxygenated perfluorinated compositions. Further applications within the scope of this invention will occur to those skilled in the art.

Claims

WE CLAIM:

1. A method of inhibiting the growth of anaer- c -s_f said method comprising contacting said anaer¬ obes with a composition comprising one or more polyfluorinated organic compounds, in which composi¬ tion oxygen has been dissolved.

2. The method of Claim 1 wherein said composi¬ tion comprises at least one compound in which 50% or more of the possible substitution sites are occupied by fluorine.

3. The method of Claim 1 wherein said composi¬ tion comprises at least one perfluorocompound.

4. The method of Claim 1 wherein said perfluoro¬ compound is selected from the group consisting of perfluorodecalin, perfluorotrimethylbicyclo[3.3.1]- nonane, perfluoro-N,N-dimethylcyclohexylmethylamine, perfluoro-2,2,4,4-tetramethylpentane, perfluoro-N- methyldecahydroquinoline, perfluoro-1-methy1-octa- hydroquinolizine, perfluoro-N-cyclohexylpyrrolidine, perfluoro-1,3,5,7-tetramethyladamantane,

1,2-di(perfluoro-n-butyl)ethylene, perfluoro¬ butyltetrahydrofurane, perfluoro-1-bromo-rι-octane, perfluorotri(n-butyl)amine, perfluorotri(n-propyl)- amine, perfluoro-l,4-dibromo-rι-butane, perfluoro-1,- 2-dibromoethane, perfluoro-1-bromocyclohexane, perfluoro-1,2-dibromocyclohexane, perfluoro-1,2-di- bromopropane, perfluoro-1-bromo-n-hexane, dichlorodi- fluoromethane, chlorotrifluoromethane, chlorodi- fluoromethane, trifluoromethane, 1,1-difluorotetra¬ chloroethane, 1,2-difluorotetrachloroethane, 1,1,1- trichlorotrifluoroethane, 1,2-dichlorotetrafluo- roethane, perfluoro-1-chloroethane, perfluoro- cyclobutane, dibromodifluoromethane, bromotrifluoro¬ methane, perfluorocyclohexane, perfluoro(methylcyclo¬ hexane) ,and perfluorotoluene.

5. The method of Claim 1 wherein said composi¬ tion comprises Fluosol-DA, FC-72, FC-84, FC-77, FC-104, FC-75, FC-40, FC-43, FC-70, FC-71, FC-80 or FC-47.

6. The method of Claim 1 wherein said composition is in the form of a mixture.

7. The method of Claim 1 wherein said composition is in the form of a suspension.

8. The method of Claim 1 wherein said composition is in the form of a solution.

9. The method of Claim 1 wherein said composition is in the form of an emulsion.

10. The method of Claim 1 wherein said composition is in the form of an aerosol.

11. The method of Claim 1 wherein said anaerobes are found in or on a living body, and said composition is non-toxic to said body.

12. A method of treating a pathogenic state in a living body, wherein said state is associated with the presence of anaerobes in the tissue of said body, said method comprising contacting said tissue with a composition according to Claim 1.

13. The method of Claim 11, wherein said patho¬ genic state is chronic inflammatory periodontal disease.