Solid substrates and methods for their preparation are provided, where enhanced functionalization of solid substrates is achieved, so that higher levels of binding of a wide variety of moieties can be obtained. The surface is nitrated with a nitronium agent, where the nitro groups may be modified in a variety of ways to serve as sites for linking. The resulting solid substrates find use in therapy, diagnosis and processing.

Novel highly functionalized solid substrates are provided in conjunction with methods for their preparation and conjugation of the functionalities to a wide variety of compositions. The subject articles find extensive use in diagnosis, therapy, processing, as well as other applications, where binding of a particular moiety to a solid surface is desired.
Solid surfaces play a significant role in the ability to separate a component from a mixture of components, to localize one or more moieties to a specific area, to investigate characteristics of various molecules, molecular aggregations, cells or tissue, or in other situations where having a moiety in a defined region, optionally in relation to a particular medium, is of interest. Thus, solid substrates find application in diagnostics, for haptens, antigens and nucleic acids, affinity chromatography, apheresis, cytophoresis as well as other applications. There are many characteristics of interest with the solid substrate, such as degree of non-specific binding, clarity, ease of fabrication, availability, etc. The characteristics are of varying importance, depending upon the particular application. For purposes of the subject invention essential characteristics include the ease with which functionalities may be introduced, the convenience of such functionalties for linking other groups, the interaction between such functionalities and the moieties attached to such functionalities, the density of functionalization available for linkage, reproducibility of the functionalization and the methods employed for linkage. There is also an interest in being able to easily view the contents of a container, such as a bottle or flask.
In many situations, it is highly desirable to have a high density of a particular moiety on the surface. By having a high density a greater number of interactions can be achieved in a predetermined surface area. This allows for smaller volumes to be used, shorter contact times, greater intensity of signal, as well as other advantages.
There is, therefore, a continuing interest in being able to develop new methods and compositions which allow for improvements of solid substrates. Such improvements can have affects on the sensitivity and efficiency of diagnostic assays, the efficiency of separation of components in a complex mixture, the ability to pack particular moieties into a small surface area, as well as other improvements.
Relevant Literature
Olah et al., J. Am. Chem. Soc. (1962) 84:3687 reports nitration in tetramethylenesulfone. Functionalization of polystyrene is described in U.S. Pat. Nos. 3,956,219; 3,886,486; 3,974,110; 3,995,094; and 4,226,958. Linking through a Schiff""s base to a solid support is described in U.S. Pat. Nos. 4,419,444 and 4,217,338.
Novel functionalized solid substrates comprising addition polymerized aromatic monomers are provided. The novel substrates are polynitrilo substituted substrates produced by nitration of the aromatic polymer surface by an electrophilic agent, particularly a nitrating agent, in a solvent substantially incapable of dissolving the polymer, containing a small amount of a polar coordinating agent. A high proportion of the aromatic groups are substituted with nitro groups. The nitro groups may then be reduced to nitrilo groups, particularly amino groups or other reduced nitrogen groups which may serve as a site for linking to a wide variety of molecules. The resulting solid substrates find particular use for separations or high density packing of a moiety of interest.
Novel functionalized aromatic substrates are provided, where substantially the entire surface of a solid aromatic substrate is functionalized. Highly functionalized surfaces are achieved employing electrophilic agents capable of aromatic electrophilic substitution, particularly a nitrating agent, and a solvent which is relatively non-polar and in which the polymer is substantially immiscible and not swelled. Included in the reaction medium is a small amount of a polar substance, particularly a hydroxylic compound, which can coordinate with the electrophilic agent.
The-solid substrate will be a homo- or co-polymer, usually non-cross-linked or having a low level of cross-links ( less than 2.0% polyene monomer) wherein at least 50 mol %, more usually at least about 80 mol % will be aromatic addition polymerizable hydrocarbon monomers, particularly having aryl groups of from 1 to 3 rings and having from 0 to 1 alkyl substituents, from 1 to 3 carbon atoms, usually at other than the para-position. For the most part, the substrate will be polystyrene which is produced in accordance with conventional ways and appears in a variety of forms, such as vials, microtiter plates, Petri dishes, plates, containers, fabricated articles, and the like. Instead of containers, the solid substrate may be particles, hollow fibers, strands, chopped strands, membranes, or the like. The polymer may be molded, extruded, machined or the like, usually resulting in a clear relatively smooth surface.
For the most part, electrophilic agents are employed, particularly nitrating agents, where the nitronium ion is coordinated with an anion which allows for significant ionization in the solvent medium, particularly a complex anion, more particularly a polyhalide anion. Illustrative polyhalide anions include tetrafluoroborate, antimony hexafluoride, phosphorous hexafluoride, etc.
The solvent will be one in which the polymer is neither soluble nor swelled, particularly tetramethylenesulfone (TMS). Included in the solvent will be a polar coordinating compound, which is able to coordinate with a nitronium ion. Conveniently, water may be used, but other oxy compounds may also find use, such as methanol. The polar compound should be capable of displacing both the anion and the solvent from coordination with the nitronium ion. The polar component will preferably be present in not greater than about equimolar amount to the nitronium ion concentration, usually present in at least about 0.05, more usually in at least about 0.25, and generally at least about 0.5 times the molar concentration of nitronium ion.
The nitronium ion will generally be at a concentration of at least about 0.05 M, usually at least about 0.1 M, and usually not greater than about 0.8 M. The particular concentration is primarily one of convenience, so long as there is a sufficient amount of reactant to substantially completely functionalize the polymeric surface.
The temperature of the reaction will vary with the nature of the reagent, generally being at least about 4xc2x0 C., and not more than about 60xc2x0 C., usually not more than about 40xc2x0 C., preferably ranging from about 30xc2x0 C. to 40xc2x0 C. The time for the reaction will vary with the temperature, so that longer times will be related to lower temperatures. Generally, the reaction will take at least 1 hr, and not more than about 48 hrs, generally running from about 2 to 24 hrs.
The reaction medium is contacted with the polymeric surface to provide an amount of reactant stoichiometric with the available aromatic groups at the surface, usually at least about 1.5 excess, where greater molar excess may be employed as convenient. In some instances it may be desirable to prepare a number of reaction mixtures using differing amounts of nitronium ion to optimize the level of functionalization of the surface.
After completion of the reaction, the reaction mixture may be discarded and the surface washed to remove any residual materials. Various solvents may be used for cleaning the surface, such as methanol, ethanol, dimethylsulfoxide (DMSO), water, nitromethane or the like. The particular material employed for cleaning is not critical to this invention and is primarily one of convenience, so long as the polymer is insoluble and is not swelled by the solvent.
The resulting polymeric surface is found by electron spectroscopy for chemical analysis (ESCA) to have a ratio of nitrogen to aryl of at least 0.1, usually at least about 0.15, more usually at least about 0.25 to about 100 xc3x85 depth. Preferably the ratio of nitrogen to aryl will be from about 0.25 to 1.
The resulting surface nitrated polyaryl solid substrate has a number of desirable characteristics. Prepared in accordance with the above procedure, the resulting functionalized polymer is clear, so that it can be used for the transmission of light through the polymer without significant interference. Thus, the nitrated polyaryl solid substrate can be used for various applications, particularly where one wishes to transmit light through a sample and have the light traverse the container, so as to determine the amount of light transmitted through the container. Or, one may wish to observe events occurring in the container, such as cell proliferation, plaque formation, cell viability, agglutination, etc. For these purposes, the nitro group will usually be modified to provide a more convenient functionality for linking various compounds.
In these applications, one is usually concerned with the effect of irradiation on a sample, where a sample may absorb light, fluoresce, or alternatively, a chemiluminescent reaction may be involved, where the sample will emit light.
The nitro groups are capable of undergoing reactions through intermediate reduced states to provide a nitrilo group such as nitroso, oxime, azoxy, azo, hydrazino or amino, depending upon the particular conditions which are employed. In some instances where the product has two nitrogens, the nitrated surface may be reacted with a second nitro compound, so that a mixed product will be obtained connected by a nitrogen bridge having from one to two nitrogen atoms, or, alternatively, an amino compound may be employed which can react with an intermediate partially reduced functionality, such as nitroso, where the nitroso group may react with the amino group to form an azo group. Of particular interest are the diaza (xe2x80x94Nxe2x80x94Nxe2x80x94) groups, such as diazo, hydrazone, semicarbazone, hydrazino, etc.
Most of the conventional reagents employed for reducing nitro groups can be employed with the nitrated substrates of the subject invention. These reducing agents include catalytic hydrogen, e.g. with platinum or palladium; stannous chloride; sodium dithionite; metal, e.g. zinc, iron, or tin-acid or -water couples; Al(Hg)-aq. ethanol; aluminum hydrides; borohydrides; ammonium sulfide; sodium arsenite; electrolytic reduction; etc.
Depending upon the nature of the functionality, various reactions may be carried out to link compounds of interest to the solid substrate surface, while still retaining clarity of the solid substrate. The reaction of nitroso groups with amines has already been indicated. Aryl amines are known to undergo a wide variety of reactions. The aryl amines may be acylated under a variety of conditions, using the non-oxo-carbonyl (carboxy carbonyl of functionality) such as acyl halides, anhydrides, active esters, such as paranitrophenyl, N-hydroxy succinimidyl, p-nitro-o-chloro-phenyl, etc., or carboxylic acids activated with activating agents such as carbodiimides.
The amino group may be reacted with a nitroso compound to form an azo linkage. Amino halides may be employed to form hydrazines with the amino group. Aldehydes or ketones may be reacted, particularly under reducing conditions, e.g. sodium borohydride, to form imines or alkylated amines, respectively.
Amines may be diazotized, where the amines may be substituted with a wide variety of functional groups, such as halides, acyl groups, oxy groups, mercapto groups, cyano groups, silyl groups , or the like.
Alternatively, the azo group may be coupled with an activated aromatic compound, such as a phenolic or a N,N-dialkylaniline compound. Additionally, tyrosinyl compounds may be coupled with diazo groups, where the tyrosine may be part of a larger molecule, such as an oligopeptide or protein. Instead of tyrosine, various purines or pyrimidines may be coupled with the azo group, where the purines or pyrimidines may be monomeric or part of RNA or DNA single stranded or double stranded chains.
Furthermore, the amino group may be functionalized to provide various reactive functionalities, such as isocyanate, urethanes, cyanamides, or the like, where the functionality may be reacted with alcohols, amines, or thiols to form urethanes and ureas.
Instead of amino groups, hydrazino groups may be prepared as indicated, or alternatively by the reduction of the diazo group, where the hydrazino group may also serve as a site for linking. Hydrazino groups can be oxidized with a variety of agents to form diazo groups or with nitrogen oxides to form azido, where the azido group under UV-irradiation forms nitrene. The highly reactive nitrenes will insert into a wide variety of compounds, particularly unsaturated compounds, both aliphatic and aromatic. Thus, aziridines may be formed which may serve as a site for bonding of a compound to the substrate.
Because of the significant interest in coupling peptides to the surface, a number of specific linking groups or spacer arms will be preferred for linking the peptide or other moiety to the surface. One group of linking group reagents will involve carboxyl groups resulting in amide formation. The carboxylic acids will be functionalized with a variety of active functionalities to produce covalent bonds with peptides, as they naturally exist or have been modified to provide a coupling site.
The functionalities for linking include aldehyde, carboxyester, haloacetamido, mercapto, and dithioether. The carboxylic acid group may be further substituted with oxy groups, hydroxy or ether, cyano, substituted amino free of hydrogen atoms, quaternary ammonium, etc. The hydrocarbon chain may be aliphatic or aromatic.
Specific linking reagents include p-formyl-benzoic acid, N-succinimidyl tartarate, mono-hydrazide of tartaric acid, p-iodoacetamidobenzoic acid, 3-mercaptopropionic acid, and 5-xcex1-pyridyl-3-dithio-propionic acid.
Reagents other than carboxylic acids may be employed to provide stable linkages, such as poly(dial-dehydes), e.g. polyglutaraldehyde, where the amino group adds by Michael addition, leaving aldehyde and olefin groups for further reaction.
In addition, the amino group may be efficiently diazotized and the resulting diazo group used in a variety of ways. Diazo groups may be substituted with oxy or thio functionalities, to produce functionalities which may be linked to a variety of groups, may be reacted with nucleophilic functionalities such as phenols, anilines, amino, etc, to form a bridge, or may be reduced to a hydrazino or amino group. The bridge may be relatively short or long depending on the purpose for the support.
Illustrative linking groups include bis-p-hydroxybenzamide of diaminopolyethyleneglycol, where one phenolic group is coupled to the diazo group and the other phenolic group may be coupled to nucleic acids or proteins through a p-di(diazo)benzene or p,pxe2x80x2-di(diazo)biphenyl. The diazo group may be directly coupled to a protein through a tyrosyl group. The phenolic group may be extended with ethylene oxide to provide a linking group, which may terminate in an amino functionality. This bridge may find application in situ peptide synthesis. The hydrazino group may be coupled, as may an amino group, to a glycol cleaved glycoprotein, to form an imine or Schiff""s base, which may be reduced to a methylenehydrazine or -amine. The hydrazine may be coupled with glyoxylic acid, leaving the carboxyl functionality for linking.
The mercapto group may provide a thiol for linking or the diazo group may be substituted by a mercapto group to form a thioether, such as reacting the diazo group with dithioerythritol.
Other bridging or linking groups employing different synthetic scenarios may also be employed. The literature has an extensive variety of reagents and procedures for coupling molecules, such as proteins, lipids, saccharides, nucleic acids and small naturally occurring or synthetic molecules to functionalities on a surface.
The bridging or linking groups may be a single bifunctional group, an oligomer or polymer, such as poly(amino acid), polyurethane, polyurea, polyalkylene oxide, polyester, polyamide, nucleic acid, or the like. In some situations it may be desirable to have a cleavable linkage, so that a bound molecule may be freed from the surface. Linkages may be employed which are cleaved by reduction, oxidation, hydrolysis, or the like, conveniently employing enzymatic catalysis. The linking group may therefore include specific recognition sites for such enzymes as dipeptidases, e.g. KEX1, collagenase, papain, neuraminidase, amylase, other endosaccharidases, endonucleases, phosphatase, pyrophosphatase, where the linkage involves an amide, ether, phosphate ester, or the like. Chemically cleavable bonds include disulfides, peroxides, acetals, azo, carboxylate ester, etc. The cleavable linking group will be selected to avoid the presence of the same or similar cleavable groups present in the moiety to be released.
The subject products, by being coupled to a wide variety of substances, can find extensive applications in diagnosis, therapy, processing, synthesis, and the like. The various groups of interest may be linked directly to the functionality bound to the aryl carbon atom or a spacer or linker arm may be used. The spacers find use where steric hindrance may be of concern, where separation of the group from the solid substrate is of interest, or where the spacing may provide an environment of interest for the group, such as polarity, charge, or the like. Usually, linking groups will be from about 1 to 30 atoms or greater, more usually from about 1 to 20 atoms in the chain and may include, carbon, oxygen, nitrogen, sulfur, phosphorus or the like. The particular choice of spacer or linking arm will depend on the functional groups involved in the linking arm, the properties of interest, synthetic convenience, spacer length, and the like.
As already indicated, a wide variety of molecules having different functions may be linked to the surface, effectively in a monomolecular layer, or polymolecular layer, if desired, and used in a wide variety of applications. The subject techniques provide for a desirable orientation of the molecules bound to the surface, in many situations enhancing their availability for binding and/or manipulation.
The compounds which are linked include ligands, such as haptens and antigens, receptors, such as antibodies, enzymes, naturally occurring receptors, such as surface membrane proteins, lectins, blood proteins, e.g. thyroxine binding globulin, complexing agents, such as cryptands, crown ethers, porphyrins, phthalocyanines, etc., nucleic acids, both RNA or DNA or oligonucleotides, dyes, fluorescers, chemiluminescers, enzymes, where the enzymes are used as catalysts rather than receptors, enzyme substrates, inhibitors or co-factors, or other molecules which may be of interest for detection of the same or different molecules, or used for manipulation, isolation, reaction, interaction, or other phenomena, where a result of interest is produced.
The literature has an extensive repertoire of groups which may be used to link almost any functionality through an amino functionality to a solid surface. U.S. Pat. No. 3,817,837 describes a large number of functionalities which may be used for linking proteins or haptens to amino functionalities, which disclosure is incorporated herein by reference. Other references which show linking to affinity columns, surfaces for apheresis, surfaces for diagnostics, surfaces for chromatography, surfaces for reaction and the like may be found in references cited in such catalogs as the BRL catalog, Bio-Rad catalog, Sigma catalog, Pierce catalog, Pharmacia catalog, etc. It is evident that the particular manner in which the compound of interest is bound to the surface may be varied widely. In particular situations one spacer arm may be selected over another because of particular properties.
The subject solid substrate allows for orientation of immunoglobulins, either as Fab fragments or Fabxe2x80x2 fragments, or as intact immunoglobulins. For example, the amino groups of the substrate may be iodoacetylated, followed by reaction with the available mercapto groups of the truncated immunoglobulin. Alternatively, the intact immunoglobulins may be oxidized with a glycol cleaving agent, e.g. periodate, and the resulting dialdehyde reacted with an amino group or hydrazine to form a Schiff base or imine.
Nucleic acids may be linked to the solid substrate using a polyalkylene glycol arm, where the alkylene will be of 2 to 3 carbon atoms, and the chain may be of from 2 to 100, usually 2 to 30 alkyleneoxy units. The linking group can be achieved by using the diazo functionality on the solid substrate with a diacylated xcex1,xcfx89-diaminopolyethylene glycol, e.g. m-dimethylaminobenzoyl derivative of diaminopolyethylene glycol. The dimethylamino activated aroyl group may be used as a site for linkage, where one will become covalently bonded to the diazo group, leaving the other to be used for linking to another compound. In this manner, phenylenediamine, benzidinie, or other aryl diamino compound may be diazotized to form a link between the dimethylaminoaroyl and a nucleic acid. The use of the diazo group for linking nucleic acids to the surface has already been discussed.
The applications of the subject invention will now be considered. The first use to be considered will be diagnostics. Diagnostics involving solid surfaces are for the most part referred to as heterogeneous in that they involve a separation step, though there is an increasing number of diagnostic protocols where a solid surface is involved without the necessity for a separation step. In diagnostics, there may be of interest the binding of members of a specific binding pair, comprising ligand and receptor, which bind to form a complex, where the homologous members of the specific binding pair have a high association constant for each other, usually greater than 108 /mol.
The haptens will generally be from about 150 to 5000 daltons, usually up to about 2000 daltons and may include naturally occurring hormones, naturally occurring drugs, synthetic drugs, pollutants, affector molecules, growth factors, lymphokines, amino acids, oligopeptides, chemical intermediates, nucleotides, oligonucleotides or the like. Diagnostics for such compounds may be in the detection of drugs of abuse, therapeutic dosage monitoring, health status, detection of disease, e.g. endotoxins, and the like. Proteins are of interest in a wide variety of diagnostics, such as detecting cell populations, blood type, pathogens, immune responses to pathogens, immune complexes, saccharides, naturally occurring receptors and the like. Receptors may find use in binding to haptens, proteins, other receptors, or the like, or detection of the presence of pathogens, the level of a particular protein in a physiological fluid, the presence of haptens in a wide variety of samples, such as physiological fluids, air, process streams, water, etc. Nucleic acids may also find use in the detection of proteins specifically binding to nucleic acids, complementary strands, and the like.
Of particular interest is the binding of microorganisms and cells, including viruses, prokaryotic and eukaryotic cells, unicellular and polycellular organism cells, e.g., fungi, animal, mammal, etc., or fragments thereof. Usually, these large aggregations will be non-covalently bound to the surface through specific binding pair member complexes. By having a high density of binding members bound to the surface, a cell or virus may be completed by a large number of binding pair members, providing very strong anchoring of the cell, virus, or fragment. The system may then be subjected to vigorous treatment without concern for dislodging the specifically bound entity, while non-specifically bound materials may be readily removed.
A large number of protocols exist for detecting the various analytes of interest. The protocols may involve use of a signal producing system, which involves a labeled conjugate, which may be directly or indirectly detected. These techniques may employ dyes, enzymes, enzyme substrates or co-factors, enzyme inhibitors, fluorescers, chemiluminescers, particles, or the like.
For the purposes of the present invention, the label should provide a signal related to the presence of analyte in the sample which results in the detection of electromagnetic radiation, particularly light in the ultra-violet, visible or infrared range. By use of the subject solid substrate, the light may be detected through the solid substrate, rather than requiring reflection, which is normally subject to significant error. In addition, techniques involving total internal reflection may be employed as described in U.S. Pat. No. 3,939,350. Other patents of interest, both domestic and foreign, which describe protocols of interest include U.S. Pat. Nos. 3,654,090, 3,850,752, 4,347,312, EPA 2,963, and references cited therein.
Assays can be carried out in accordance with the various protocols. In accordance with the subject invention, the sample is contacted with the subject solid substrate and various operations may be carried out, such as the addition of various reagents, incubations, washings, and the like. The final result of the assays will be the change in the amount of a product which absorbs or produces light, either by light absorption or by light emission in relation to the presence or amount of the analyte of interest. Usually, this is as a result of formation of a specific binding complex between complementary members of a specific binding pair, where one of the members may serve as a bridge to form a sandwich, or there may be a single complex, or complexes may be bound to complex binding proteins, such as S. aureus protein A, rheumatoid factor, immunoglobulins specific for immune complexes, or the like.
By having fluorescent markers, such as fluorescent particles, fluorescent conjugated antibodies, or the like, the sample may be irradiated with light absorbed by the fluorescers and the emitted light measured. For example, where one has microtiter wells, one can measure the fluorescent light emitted through the sides of the well, substantially minimizing the background resulting from the activating light. Where dyes are employed as the label or produced as a result of a reaction, e.g. an enzymatically catalyzed reaction, the light may be transmitted through the sample and the container and measured as an indication of the presence of the analyte, since there is a relatively low level of absorption by the container. Thus, highly sensitive assays can be employed with the subject system.
Similarly, with nucleic acid assays involving hybridization, one can carry out the necessary steps to determine whether complementary sequences are present, and by employing a wide variety of protocols, provide for a colored or fluorescent label or product of the label, which will indicate the presence or absence of the complementary sequence.
For example, one could activate the surface immediately prior to carrying out the assay by diazotizing the amino functionalities, add the nucleic acid sample to the activated surface, so as to be covalently bound, and then employ probes having a sequence complementary to the sequence of interest and functionalized, for example, by having a biotin label. After completion of the hybridization step, one could add enzyme conjugated to avidin, which would bind to any biotin bound to the surface through hybridization. After washing away non-specifically bound avidin, the substrate for the enzyme could be added and the formation of product would be indicative of the presence and amount of the complementary sequence.
The subject containers may also be used for detecting pathogens. Monoclonal antibodies may be linked to the surface to serve as catching antibodies. The sample would then be added and cells having the epitope recognized by the antibody would bind to the antibody on the surface. Non-specifically bound cells are washed away leaving substantially only specifically bound cells. Labeled monoclonal antibodies are then added to the container which are specific for an epitope other than the epitope recognized by the catching antibody. After incubating to allow reaction between the antibodies and cells, non-specifically bound antibodies are washed away and the presence of the label determined, as described previously.
A variation would be to employ an antigen recognized by a cell receptor. The antigen would be bound to the surface to catch the cells and a labeled antigen used to label the cells. The receptor could be surface immunoglobulin (sIg). In this way the presence of the specifically bound cells could be determined, whereby having the antigen of interest complementary to the receptor bound to the surface, cells having the sIg specific for such antigen could be determined. Instead of having antigen, one would have antibodies to the antigen bound to the surface to non-covalently bind the antigen to the surface.
The subject solid substrates can also be used for affinity columns, chromatography, or the like, where a particular substance is bound to the surface and complex formation or other binding event can be detected. With the subject solid substrates, one can visually observe along the length of a column and determine the particular site at which complex formation occurs. This technique can also be used for diagnosis, where different antigens or receptors may be bound at different sites along the column, followed by introducing the sample through the column, followed by adding the signal producing system, e.g. enzyme bound specific binding pair member and developer, and detecting the areas of color formation.
The subject solid substrates may also find use in isolating various products of interest, such as blood plasma proteins, growth factors, clotting factors, anti-clotting factors, or the like, which may then be released from the complex by various salt solutions.
The solid substrates may be used for a variety of other purposes, whenever one wishes to provide a high density of oriented molecules at a surface or visualize events or provide for ready transmission of light, where a substance is non-diffusively bound to a solid surface.

Activated and conjugated polystyrene substrate


The present invention discloses a light switch designed physically similarly to a present day conventional light switch having a face plate and on/off switching capability for connection to standard house current using 110/115 volts AC electricity. The present invention discloses the light switch having memory capability wherein the light usage patterns actually used by the household resident can be stored into memory so that when the household residents are away from home they can activate the memory of the present invention so that the light switches are operated in their absence according to the actual usage pattern which has been exhibited by them while present in the house.

1. Field of the Invention
The present invention relates generally to light switches, and, more particularly, is concerned with a light switch having memory capability for storing light usage patterns therein.
2. Description of the Prior Art
Light switches have been described in the prior art. However, none of the prior art devices disclose the unique features of the present invention. It is felt that a light switch capable of storing usage patterns would be useful.
In U.S. Pat. No. 5,160,853, dated Nov. 3, 1992, Simon, et al., disclosed a programmable timer for turning a light on and off in response to changing sunset, daylight savings time, and other programmable conditions. The programmable timer includes an input device, a microprocessor, and a switch. The input device provides calendar, geographical, and daylight savings information to the microprocessor, which stores the programming information. The microprocessor computes an effective switching time from the stored information. The microprocessor employs program logic that compares the effective switching time to the current time to generate a timing control signal. The timing control signal, in turn, causes the switch, to turn a light on and off. In the preferred embodiment, the microprocessor and the switch are included within a housing which mounts on a wall in place of a normal light switch. A liquid crystal display is disposed on a face of the housing to provide a read out of the programming information and of the time of day. The input device is disposed on the face of the housing and includes a key matrix having a plurality of finger settable key type programming switches. Information is programmed by depressing each key while the programming logic is in a particular program mode.
In U.S. Pat. No. 5,465,031, dated Nov. 7, 1995, Nilssen disclosed a self-contained programmable actuator which can easily be mounted directly onto the outside of the face plate of a standard wall switch having a built-in light dimmer. This actuator can be programmed to adjust the position of the dimmer""s mechanical control input in accordance with a program that automatically repeats on a diurnal, weekly or other cyclical basis. The actuator includes a small battery, a miniature electric motor with a gear/linkage mechanism operable to engage with and to move the dimmers mechanical control input over its complete range, and a quartz clock-based programming means having programming input keys and time display means.
In U.S. Pat. No. 5,629,587, dated May 13, 1997, Gray, et al., disclosed a programmable lighting control system for advertising, decorative, artistic and Christmas lighting applications, which consists of a standalone controller, an optional power booster device, and a personal computer compatible software program. The controller receives power via a standard AC outlet receptacle and includes: a plurality of AC output receptacles for connection to either series or parallel connected Christmas tree type lights or the like; a microcontroller to provide timing and control signals that are applied to solid state switching devices to drive the outlet receptacles; a non volatile memory to store custom user defined lighting sequences; a rotary switch to enable the selection of either pre-programmed sequences or user defined sequences: and a serial communication port. The personal computer compatible software program enables the user to create custom lighting sequences, which can be downloaded to the light controller non-volatile memory via the serial port. The optional power booster device can be used to increase the output power capability of each of the individual controller output circuits.
In U.S. Pat. No. 4,279,012, dated Jul. 14, 1981, Beckedorff, et al., disclosed a programmable apparatus for providing random on/off control of electrical devices such as appliances. Such control may be programmed for an entire week, with different programming for each day, and with programming control over small blocks of time, e.g., thirty minutes. Programming is secured by the use of a combination or lock and, in addition, the AC line cord of the electrical device, which receives or does not receive AC power depending upon the status of the program, is locked into the apparatus. During operation, the program in the apparatus may be interrogated and displayed; otherwise, the time of day is displayed. The apparatus also includes a look-ahead feature, turning on an appliance, such as a television set, earlier than programmed in order to provide sufficient warm-up time.
In U.S. Pat. No. 4,344,000, dated Aug. 10, 1982, Schornack, et al., disclosed one form of a timer which is designed to be mounted over and behind a conventional wall switch cover plate and includes a two position time condition-setting arm, a rotatable and depressible time setting dial knob and an indicator on the front of the cover plate. When the arm is moved to a timer-on position, the indicator flashes to indicate that timer programming is needed. This may be achieved in real time over the first twenty-four hours after a 30 minute play period by depressing the knob acting as an on and off power switch control push button to turn the light circuit on and off in a normal manner. Fast normal programming is achieved by rotating the knob to various time settings and depressing the knob to se ON or OFF markers in timer memory. A full twenty-four hour programming is completed by either completing a full rotation of the knob over the full twenty-four hour time settings, which at least one ON and one OFF depression, or by a similar pair of knob depressions without such a full rotation, OFF markers then being automatically set by the time to complete the timer programming. The indicator comes steady on to indicate that the timer is fully programmed and in an automatic mode of operation. The automatic operation is temporarily overridden by depression of the knob or permanently overridden by a simple distinctive speed or time duration depression of the knob.
In U.S. Pat. No. 4,354,120, dated Oct. 12, 1982, Schornack disclosed an electrical timer which automatically actuates an output electrical load device such as electric light according to a program stored as markers in a memory unit over a 24-hour interval, whereupon the program is recycled. By actuating one or more electric lights within an empty dwelling, an illusion of occupancy is thus conveyed to an observer. In the preferred form of the invention the memory unit is a recirculating shift register. A daily variability of the timer output is provided to prevent precise daily replication of the load device duty profile. One version causes the program output to be delayed by 15 minutes on alternate days by alternately taking the output from the last or the next-to-last storage element of the shift register. The stored duty profile is in 15-minute blocks. A second version uses 30-minute blocks and minimizes possible output distortion during the transition from normal to delayed display by using two timing pulses 16 minutes apart to display the program output on alternate days 8 minutes early or 8 minutes late. The second version allows use of a smaller amount of memory storage.
In U.S. Pat. No. 4,570,215, dated Feb. 11, 1986, Chan disclosed a fully automatic programmable wall switch timer for controlling lighting circuits. The device provides user selected lighting circuit operation at predetermined event times, for predetermined event duration at said times, and at a selected light brightness or intensity level. The device is microprocessor controlled and is intended for installation in a standard wall switch box using existing wall switch plates. Included are a time of day and programming display and a three-way switch option. Separate switches are provided for isolating the unit from the power line, selecting lighting circuit operation, and advancing the display time. Additionally, various user-programming modes are provided by operation of said switches and in conjunction with each other.
While these automatic light switches may be suitable for the purposes for which they were designed, they would not be as suitable for the purposes of the present invention, as hereinafter described.
The present invention discloses a light switch designed physically similarly to a present day conventional light switch having a face plate and on/off switching capability for connection to standard house current using 110/115 volts AC electricity. The present invention discloses the light switch having memory capability wherein the light usage patterns actually used by the household resident can be stored into memory so that when the household residents are away from home they can activate the memory of the present invention so that the light switches are operated in their absence according to the actual usage pattern which has been exhibited by them while present in the house.
An object of the present invention is to store home light usage patterns of residents for later use when the residents are away from home. A further object of the present invention is to provide improved home security so the actual light usage pattern of the residents is used when the residents are away in order to deter thieves from the home. A further object of the present invention is to provide a timer for light usage when the residents are away from the home.
The foregoing and other objects and advantages will appear from the description to follow. In the description reference is made to the accompanying drawings, which form a part hereof, and in which is shown by way of illustration specific embodiments in which the invention may be practiced. These embodiments will be described in sufficient detail to enable those skilled in the art to practice the invention, and it is to be understood that other embodiments may be utilized and that structural changes may be made without departing from the scope of the invention. In the accompanying drawings, like reference characters designate the same or similar parts throughout the several views.


A method and composition for the treatment of musculoskeletal disorders in mammals by the application of a topical composition comprising a permeation enhancing amount of one or more penetration enhancers, and one or more bio-affecting agents to provide anti-inflammatory relief and analgesia to the applied body part.

A method and topical composition for the treatment of musculoskeletal disorders.
The musculoskeletal system consists of bones, muscles and joints. Ten percent of medical visits to physicians are for disorders of the musculoskeletal system. Musculoskeletal disorder include: sprains, strains, tendinitis, tenosynovitis, fibromyalgia, osteoarthritis, rheumatoid arthritis, gout, pseudogout (calcium pyrophosphate deposition disease), polymyalgia rheumatica, bursitis, acute and chronic back pain and osteoporosis, which interfere with the normal performance of activities of daily living. Injuries include sprains, strains and tears of ligaments, tendons, muscles and cartilage damage. Pain is the most common symptom and is frequently caused by injury or inflammation. Besides pain, other symptoms such as stiffness, tenderness, weakness and swelling or deformity of affected parts are manifestations of musculoskeletal disorders. Sports injuries are a significant cause of musculoskeletal disorders resulting in pain, strain, sprains, stiffness and leg cramps.
Occupational injuries however, have become this country""s most costly form of illnesses. The Bureau of Labor Statistics reported in 1992, that one half of the 2.3 million nonfatal occupational injuries and illnesses which resulted in days away from work involved musculoskeletal disorders. The Occupational Safety and Health Administration (OSHA) estimates in 1999, that more than 647,000 Americans suffer from injuries or illnesses due to work-related musculoskeletal disorders (WMSDs). These disorders account for more than 34% of all workdays lost to injuries and illnesses and cost employers $15 to $20 billion per year in direct workers"" compensation costs and another $100 billion on lost productivity, employee turnover, and other indirect expenses. Cumulative trauma disorders (CTDs) frequently involve the upper extremities, such as wrists, shoulders or elbows. Carpal tunnel syndrome of the wrist has become the fastest growing occupational hazard in the United States today.
Arthritis, a musculoskeletal disorder, is the leading cause of disability in the United States. The Centers for Disease Control and Prevention (CDC) stated that arthritis and other rheumatic conditions accounted for about 744,000 hospitalizations and 4 million days of care in 1997. It is estimated that 43 million people are now affected and expected to increase to 60 million people by 2020. Arthritis costs the country about $65 billion a year.
Osteoporosis afflicts 200 million people worldwide and 25 million people in the U.S., of whom 80% are women, yet one man in three will also get osteoporosis. One woman in two over the age of 60 is likely to have an osteoporotic bone fracture. The incidence of hip fracture exceeds that of cancer of the breast, cervix and uterus combined. Sixteen percent of patients suffering hip fractures will die within six months, while 50% will require long-term nursing care. The estimated cost of treatment and care for osteoporosis and related fractures exceeds 10 billion per year in the U.S. alone.
Treatment usually consists of an oral ingestion of an analgesic such as nonsteroidal anti-inflammatory drugs (NSAIDs). Adverse side effects of oral NSAIDs, such as hypersensitivity, gastropathy, renal impairment, liver toxicity and prolonged bleeding merit concern in the very young and elderly. The use of topical analgesic compositions to treat musculoskeletal disorders is an effort to overcome the side effects of oral preparations with the advantage of delivering the analgesic directly to the affected body part.
This invention relates to a method for the treatment of musculoskeletal disorders in mammals by the application of a topical composition comprising a permeation enhancing amount of one or more penetration enhancers, and one or more bio-affecting agents to provide anti-inflammatory relief and analgesia to the applied body part.
The components of the method and composition of the present invention will be discussed separately because no prior art embraces them as a solitary method or composition to reduce inflammation and provide analgesia to the affected body part due to musculoskeletal disorders. The topical composition of the present invention comprises one or more penetration enhancers and one or more bio-affective agents.
A penetration enhancer or permeation enhancer is an agent used to increase the permeability of the skin to a pharmacologically active agent to increase the rate at which the drug diffuses through the skin and enters the tissues and bloodstream. A chemical skin penetration enhancer increases skin permeability by reversibly altering the physiochemical nature of the stratum corneum to reduce its diffusional resistance. In a review of the technical and patent literature up to 1996, more than 275 different chemical compounds were found to be cited as skin penetration enhancers. Most of the compounds are generally recognized as safe (GRAS) ingredients that would often be considered inert by a formulator. Osborne D W, Henke J J, Pharmaceutical Technology, November. 1997, pp 58-86. The compounds cited in the article are incorporated by reference. Examples of penetration enhancers include: alcohols, such as ethanol and isopropanol, polyols, such as n-alkanols, limonene, terpenes, dioxolane, propylene glycol, ethylene glycol, other glycols, and glycerol; sulfoxides, such as dimethylsulfoxide (DMSO), dimethylformanide, methyl dodecyl sulfoxide, dimethylacetamide; esters, such as isopropyl myristate/palmitate, ethyl acetate, butyl acetate, methyl proprionate, capric/caprylic triglycerides, ketones; amides, such as acetamide oleates such as triolein; various surfactants, such as sodium lauryl sulfate; various alkanoic acids such as caprylic acid; lactam compounds, such as azone; alkanols, such as oleyl alcohol; dialkylamino acetates, and admixtures thereof.
A number of patents disclose the use of penetration enhancers to deliver medications transdermally. Grasela et al, U.S. Pat. No. 5,837,289, discloses the use of at least two separate penetration enhancers in a cream to deliver an extensive list of medications, Catz et al, U.S. Pat. No. 5,238,933, discloses a skin permeation enhancer composition comprising a lower aliphatic ester of a lower aliphatic carboxyl acid in combination with a lower alkanol to administer an active agent. Sharma et al, U.S. Pat. No. 5,229,130, discloses a vegetable oil-based skin permeation enhancer to deliver active agents through the skin. Tsuk, U.S. Pat. No. 4,933,184, discloses a transdermal composition that uses methanol either sequentially or simultaneously to deliver drugs. Havemeyer et al, U.S. Pat. No. 4,342,784, discloses a method of topically administering a gel with DMSO and carboxy polymethylene resin with a neutralizing agent to enable the skin salt to break down the gel to release the DMSO. Rajadhyaksha, U.S. Pat. No. 5,482,965, discloses a transdermal composition containing a dioxane. Samour, U.S. Pat. Nos. 5,620,980, 5,807,957, discloses the use of a dioxolane and urethane to treat hair loss. None of the above cited patents teach or suggest the use of the method or composition outlined in the present invention.
The present invention relates to novel compositions for topical application and delivery of bio-affecting agents through the protective outer layer of the skin, into the underlying tissues and into the vascular network of the targeted body part. The term xe2x80x9cbio-affecting agentxe2x80x9d refers to any chemical substance or formulation which beneficially affects the body. The bio-affecting agents of the preferred composition comprises; analgesics, NSAIDs, anti-inflammatory agents, colchicine, S-adenosyl-methionine, methyl-sulfonyl-methane, antioxidants, anti-infectives, zinc compounds, amino sugars, aloe vera extracts, and other active agents to form a solution, suspension, cream, ointment, gel, film, or spray.
The concentration of the bio-affecting agents in the composition can also vary greatly and will be dependent upon may factors, e.g. type, bioavailability, potency, surface area to which it is applied, composition used and the amount of the penetrating agents used. The concentration of bio-affecting agents will vary from about 0.1 % to 25 % of the total composition, and may be suspended or dissolved.
A wide variety of therapeutic agents, known to provide beneficial effects when absorbed into the tissues and vascular network, in combination with a penetration enhancer, facilitates penetration through the skin and absorbed into the vascular network of the targeted body parts. This topical administration offers a significant advantage over oral administration of therapeutic agents by overcoming the difficulty of poor gastrointestinal absorption, by using a lower dosage than required orally, and allows more of the active agents to provide therapeutic relief Bio-affective agents to be used in the composition includes the following:
Pain is usually the presenting symptom of musculoskeletal disorders. The pain response is a protective reflex system warning an individual of tissue injury. Most commercial topical analgesics use a counter-irritant, such as methyl salicylate, menthol, camphor, eucalyptol and derivatives or mixtures thereof, or rubefacients, such as capsicum, oleoresin chloroform and the like, formulated as an ointment or gel.
The use of counter-irritants and rubefacients to achieve analgesia are well known in the art. Arora, U.S. Pat. No. 5,223,257, discloses an analgesic composition of methyl salicylate, olive oil, eucalyptus oil and isopropyl alcohol. Nichols, U.S. Pat. No. 5,223,267, discloses an analgesic composition of cellulosic powder, counter-irritant (salicylates, menthol, comphor, eucalyptol), analgesic (aspirin, triethanolamine salicylate, ibuprofen), steroid (hydrocortisone), mineral oil, emollient and alcohol. Fisher, U.S. Pat. No. 3,880,996, discloses a preparation of salicylate, menthol, polysiloxane and a vasodilator, such as histamine. Beck, U.S. Pat. No. 5,073,366, discloses a composition containing camphor and eucalyptus oil. Elden, U.S. Pat. No. 5,814,659, discloses a topical composition of a lidocaine analgesic, benzyl alcohol, urea, fatty acid, emulsifier, gel, preservative and organic base. Saitoh et al, U.S. Pat. No. 4,775,667, discloses a topical composition of ethylene glycol monosalicilate, methanol and a small amount of corticosteroid. Hosick, U.S. Pat. No. 4,120,976, discloses the use of methylenedioxyamphetamine to treat arthritis. None of the above cited patents teach or suggest the use of the method and composition outlined in the present invention.
Nonsteroidal anti-inflammatory agents (NSAIDs) are also useful in relieving pain and tissue swelling, chiefly by inhibiting the biosynthesis of prostaglandins. In small doses, NSAIDs have an analgesic action, but full doses have both analgesic and anti-inflammatory actions, and are effective in reducing pain and swelling. While pain relief from a headache can be obtained with a single 200-400 mg dose of ibuprofen, a full anti-inflammatory effect for bursitis might require 3,200 mg/d of the same drug. NSAIDs fall in seven major classes: proprionic acid derivatives, indole derivatives, fenamates, pyrrolealkanoic acids, pyrazolone derivatives, oxicams and salicylic acids.
One possible reason that has emerged is the selectivity of different NSAIDs even within the same class for different forms of cyclooxygenase (COX) enzymes, which are involved in the production of prostaglandins. Researchers are seeking NSAID formulations that would inhibit COX-2, which is responsible for inflammation, while sparing COX-1, which produces the more protective prostaglandins. Those NSAIDs inhibiting COX-1 most strongly are also known to have greater ulcerogenic activity. NSAIDs that are relatively COX-2 selective, such as nabumetone and etodolac, may have a slightly lower risk of GI and renal adverse effects. Ringel M, Which NSAID for acute joint pain? Patient Care, Feb. 28, 1998. Chronic use of NSAIDs in older adults is also associated with an increased risk of renal dysfunction. Field T S, J Am Geriatr Soc 1999,47:507-511.
Table 1 shows the recommended maximum daily oral dosage for some of the more popular NSAIDs and the suggested topical dosage for achieving the same anti-inflammatory benefits to the affected body part. The use of topical NSAIDs in humans and animals demonstrated a lower plasma concentration than with systemically administered drugs, but maintained the same anti-inflammatory effect. Vaile J H, Davis P, Topical NSAIDs for musculoskeletal conditions. A review of the literature. Drugs 1998 November;56(5):783-99. Topical diclofenac eye drops proved as effective as prednisone in controlling inflammation and discomfort after strabismus surgery. Apt L, Voo 1, Isenberg S J, A randomized clinical trial of the nonsteroidal eyedrop diclofenac after strabismus surgery. Ophthalmology 1998 August;105(8):1448-52. A study compared topical eltenac with oral diclofenac in osteoarthritis patients and found a similar symptomatic relief, but that the number of GI reactions were three times higher in the diclofenac group. Sandelin J, et al. Local NSAID gel (eltenac) in the treatment of osteoarthritis of the knee. A double blind study comparing eltenac with oral diclofenac and placebo gel. Scand J Rheumatol 1997;26(4):287-92.
The use of NSAIDs as analgesics and anti-inflammatory agents is well known in the art. Paradies, U.S. Pat. No. 5,604,206, discloses complexes of ibuprofen and amino sugars, but does not teach a transdermal preparation. Veronesi, U.S. Pat. No. 4,748,174, discloses a composition to improve GI absorption using meglomine or glucamine salt and an NSAID. Cavanaugh, Jr., U.S. Pat. No, 5,626,838, discloses a method to treat oral cancer with a topical composition of an NSAID. Adger et al, U.S. Pat. No. 5,811,558, discloses a process for preparing a salt of etodolac. Shunshine et al, U.S. Pat. Nos. 4,722,938, 4,780,463, and 4,923,898 all disclose a method and composition of an NSAID and a muscle relaxant. None of the above cited patents teach or suggest the use of the method and composition outlined in the present invention.
Topically applied NSAIDs are well known in the art. Inagi et al, U.S. Pat. No. 4,309,414, discloses an ointment comprising; indomethacin, glycol, alcohol, water, gelating agent and adjuvant. Kigasawa et al, U.S. Pat. No. 4,500,511, discloses a composition comprising clidanac, gelling agent, neutralizer, solubilizer, water and absorption promoter. Kamishita, U.S. Pat. No. 4,543,251, discloses a gel preparation comprising diclofenac, lower alkanol, water, glycol, carboxyvinyl polymer and aliphatic amine. Ogiso, U.S. Pat. No. 4,873,081, discloses a percutaneous absorption preparation of indomethacin, ethylene glycol, DMSO, gelatinizing agent and an absorption adjuvant. Kamiya et al, U.S. Pat. No. 4,948,588, discloses a preparation of aspirin and an absorption accelerator. Cooper et al, U.S. Pat. No. 4,954,487, discloses a topical composition of an NSAID, ethanol and penetration enhancing carrier. Betlach, II, U.S. Pat. No. 5,374,661, discloses a composition of diclofenac, thickening agent, alcohol, glycol, ether alcohol and fatty alcohol ester, water and neutralizing agent of ethoxydiglycol. None of the above cited patents teach or suggest the use of the method and composition outlined in the present invention.
Inflammation is a fundamental pathologic process involving complex reactions that occur in the affected blood vessels and adjacent tissues in response to an injury or abnormal stimulation caused by a physical, chemical, or biologic agent. The acute inflammatory response begins after cellular injury due to microorganisms, physical agents (such as burns, radiation, and trauma), chemicals, necrotic tissue, and immunological reactions. Five classic signs are manifested in acute inflammation; redness, heat, pain and loss of function. These signs are induced by changes which take place in the microvasculature (arterioles, capillaries, and venules) and the interstitial areas (fluid-filled regions between cells and tissues). These include changes in vascular flow and caliber, changes in vascular permeability, and leucocyte exudation. The first change involves vasodilation of the vessels and increased blood flow. The second change involves increased permeability of the blood vessels with a movement of fluid and proteins out of the vessels creating edema of the tissues. The final change occurs as white blood cells infiltrate and accumulate in the surrounding tissue. The increased blood flow and permeability of the microvascular system at the inflamed body part facilitates treatment to the area by using a penetration enhancer to deliver the bio-affective agents.
The spread of the acute inflammatory response following injury to a small area of tissue suggests that chemical substances are released from injured tissues, spreading outwards into uninjured areas. These chemicals, called endogenous chemical mediators, cause vasodilation, emigration of neutrophils, chemotaxis and increased vascular permeability. Histamine is a chemical mediator in acute inflammation and causes vascular dilatation and vascular permeability. It is stored in mast cells, basophil and eosinophil leucocytes, and platelets. Histamine release is stimulated by complement components C3a and C5a and by lysosomal proteins released from neutrophils. Prostaglandins are a group of long-chain fatty acids derived from arachidonic acid. They increase vascular permeability, and platelet aggregation. Drugs such as aspirin and NSAIDs inhibit one of the enzymes involved in prostaglandin synthesis. Other chemical mediators include; leukotrienes, serotonin and lymphokines. Plasma contains four enzymatic cascade systems; complement, the kinins, the coagulation factors and the fibrinolytic system.
Chronic inflammations are characterized by a longstanding dull pain, and indurated swelling, and the presence of granulation tissue. The predominant cells seen in chronic inflammation are the mononuclear leukocytes, such as macrophages, lymphocytes, and plasma cells. A fibroblastic proliferation is seen more often than a fluid exudate. Some bio-affective agents with anti-inflammatory properties are the following:
Gout is a musculoskeletal disorder and common type of arthritis caused by an increased concentration of uric acid in joints, tendons, cartilage, and surrounding soft tissues. It is one of the oldest recorded diseases in man, and afflicts the joint at the base of the big toe but can also cause pain and swelling in other joints such as knees, ankles, hands and wrists. Gout is becoming more frequent since low dose aspirin and diuretics, such as hydrochlorothiazide, are commonly used in the adult population, which inhibit renal excretion of uric acid and lead to the symptoms of gout. The acute inflammation of gout is believed to be activated after neutrophils phagocytize urate crystals with the ensuing release of chemotactic and other substances capable of mediating inflammation. Chronic gout results in an invasion of the articular and periarticular tissues, with structural derangement and secondary osteoarthritis. The inflammatory response to urate crystals requires leukocytes to be present.
Colchicine, an alkaloid, is effective in treating gout and dermatoses such as psoriasis, and Sweet""s syndrome. Sullivan T P, King LEJr, Boyd A S, Colchicine in dermatology J Am Acad Dermatol 1998 December;39(6):993-9. Colchicine inhibits neutrophil chemotaxis thus decreasing the inflammatory process. Ben-Chetrit E, Levy M, Does the lack of the P-glycoprotein efflux pump in neutrophils explain the efficacy of colchicine in familial Mediterranean fever and other inflammatory diseases?Medica/Hypotheses 1998 November;51(5):377-80. Colchicine is a microtubule depolymerizing agent and a competitive antagonist at the GABA(A) receptor. Bueno OF, Leidenheimer N.J., Colchicine inhibits GABA(A) receptors independently of microtubule depolymerization. Neuropharmacology 1998;37(3):383-90. Colchicine has no effect on uric acid levels; rather it stops the inflammatory process by inhibiting neutrophil migration into areas of inflammation, blocks the release of chemotactic factor, reduces mobility and adhesion of polymorphonuclear luekocytes, inhibits the production of leukotriene, and reduces the production of lactic acid. Eighty percent of patients are unable to tolerate an optimal dose of colchicine because of gastrointestinal side effects. Murray M, Pizzorno J, Encyclopedia of Natural Medicine 2nd Ed. 1998 Prima Publishing, Rocklin, Calif. Early degenerative changes in cartilage and synovial tissues of the knee joint are manifested by pain and limitation of motion. In a controlled study, colchicine and diclophenac were irrigated into the knee joints and after two years an examination demonstrated less fissure formation and cartilage thinning and increased chondrocyte proliferation and replication. Doral M N, et al, The Role of Irrigation with Colchicine and Diclophenac Sodium in experimental gonarthrosis. First Biennial Congress of the International Society of Arthroscopy, Knee Surgery and Orthopaedic Sports Medicine, Buenos Aires, Argentina, May 11-16, 1997.
Colchicine causes microtubule depolymerization which leads to a loss of cytokine receptor expression and underlies the anti-inflammatory actions. Rao P, et al, Colchicine down-regulates LPS-induced granulocyte-macrophage colony-stimulating factor production in murine macrophages. J Immunol 1997 October 1;159(7):3531-9. Colchicine inhibits COX-2 derived prostenoids, including prostaglandin E2 (PGE2) and thromboxane A2 (TXA2). Pouliot M, et al, Monosodium urate microcrystals induce cyclooxygenase-2 in human monocytes. Blood 1998 March;91(5):1769-76. Colchicine blocks sodium channel transport and can block axonal nerve transport but spares C-fiber nociceptor function. Kingery W S et al, Colchicine treatment of the sciatic nerve reduces neurogenic extravasation, but does not affect nociceptive thresholds or collateral sprouting in neuropathic or normal rats Pain 1998 January;74(1):11-20.
The inventor is not aware of the use of colchicine as a topical composition to treat musculoskeletal disorders. Colchicine is given orally and the recommended dosage is 1 mg orally followed by 0.5-0.6 mg every two hours until symptoms cease, not to exceed a total dosage of 6-8 mg. The suggested topical dosage is 0.2-0.5 mg.
The use of colchicine as an anti-inflammatory agent is well known in the art. Eakins et al, U.S. Pat. No. 4,190,673, discloses an ophthalmic preparation of 0.1 to 2% colchicine to treat glaucoma. Brossi et al, U.S. Pat. No. 4,533,675, discloses a colchicine compound to treat gout. Sackler et al, U.S. Pat. No. 5,747,060, discloses a formulation for local anesthesia comprising an anesthetic, controlled release material and colchicine to prolong the duration of the local anesthesia. Bombardelli et al, U.S. Pat. Nos. 5,843,910, 5,880,160, discloses colchicine derivative compounds and method to treat tumors due to antiproliforative and cytotoxic effects. NSAIDs have also been cited for treating gout. Mullane, U.S. Pat. No. 4,663,345, discloses a method for lowering uric acid using etodolac. Gertner, U.S. Pat. No. 5,061,724, discloses a method for alleviating symptoms of gouty arthritis with a bandage of skin cream and indomethacin. None of the above cited patents teach or suggest the use of the method and composition outlined in the present invention.
Methyl-sulfonyl-methane (MSM) or dimethyl sulfone is essentially DMSO with an extra oxygen molecule and lacks the lipid-solubility of DMSO, but can be coupled with another penetration enhancer. In the body, MSM gives up its sulfur to form methionine and cysteine for connective tissue. MSM is anti-inflammatory and analgesic and useful for muscle soreness and cramps, prevents cartilage degeneration and improves joint flexibility. The therapeutic dosage range for MSM is 2-10 grams orally per day. The recommended topical dosage range is 1-5 grams.
Numerous patents for MSM were filed by Herschler. U.S. Pat. Nos. 4,296,130, discloses a method for softening skin; 4,477,469 discloses a composition of MSM and carbamide to soften skin; 4,863,748 discloses a method for adding sulfur to the diet with MSM; 4,973,605 discloses a method for treating muscle cramps associated with arthritis with oral MSM; and 5,071,878 discloses a method for using MSM in a diet for sulfur and health reasons. None of the above cited patents teach or suggest the use of the method and composition outlined in the present invention.
S-adenosyl-methionine (SAMe) is the activated form of methionine. SAMe appears to counteract cytokines, protects cartilage, inhibits destructive free radicals, and may reverse the effects of homocysteine on cartilage and/or inhibit enzymes. SAMe increases the number of chondrocytes (cartilage cells) and proteoglycans (protein), and reverses the damaging effects of TNF. SAMe also has a natural antidepressant effect which is beneficial to those patients with musculoskeletal disorders.
The recommended oral dosage of SAMe is 1,200 to 1,600 mg per day. The suggested topical dosage of SAMe is 800 mg.
The use of SAMe is well known in the art. Fiecchi, U.S. Pat. No. 4,057,686, discloses the process for the preparation of salts of SAM. Stramentinoli et al, U.S. Pat. No. 4,454,122, discloses a method to treat inflammation and pain using MTA. Zappia et al, U.S. Pat. No. 4,764,603, discloses a process for preparing stable salts of SAMe. Le Fur et al, U.S. Pat. No. 4,956,173, discloses a method and composition to treat the skin using SAM. Gennari, U.S. Pat, No. 5,128,249, discloses a process for producing SAMe. Kurobe et al, U.S. Pat. No. 5,166,328, discloses a SAMe derivative useful for treating stroke. Braganza et al, U.S. Pat. No. 5,196,402, discloses a method for treating pancreatitis using SAMe. Kawabata et al, U.S. Pat. No. 5,466,678, discloses a method of reducing nephrotoxicity of cisplatin therapy using SAMe. None of the above cited patents teach or suggest the use of the method and composition outlined in the present invention.
Osteoporosis is characterized by progressive loss of bone architecture and mineralization leading too the loss of bone strength and an increased fracture rate. The skeleton is constantly being remodeled by a balance between osteoblasts, that lay down new bone, and osteoclasts, that break down or resorb bone.
Zinc plays a physiological role in the regulation of bone metabolism, by stimulating bone formation and mineralization and an inhibitory effect on bone resorption. Zinc activates aminoacyl-tRNA synthetase in osteoblastic cells, stimulates cellular protein systhesis, and inhibits osteoclast-like cell formation in marrow cells. Bone zinc content is decreased by development, with aging, skeletal unloading, and postmenopausal conditions. Zinc plays a role in the preservation of bone mass. Most zinc compounds, such as zinc sulfate, are useful for the prevention of osteoporosis, but a recent study confirmed that xcex2-Alanyl-L-histidinato zinc (AHZ) has a potent effect on bone formation and calcification. Yamaguchi M, Role of Zinc in Bone Formation and Bone Resporption, J. of Trace F. Elements and Experimental Medicine 1998;11:119-135.
Zinc compounds have anti-inflammatory and anti-infective properties. In a recent published article, Petrus E J et al., Current Therapeutic Research, 1998; 59/9: 595-607, the inventor served as chief investigator for a randomized, double-masked, placebo-controlled clinical study of the effectiveness of zinc acetate lozenges on common cold symptoms in allergy-tested subjects. Those subjects who used the zinc lozenges had both a shorter duration and severity of common cold symptoms. Those subjects who were positive for allergies, were more responsive to zinc by having a shorter duration of nasal symptoms. The study cited many references that reported the following benefits and effects of zinc compounds:
Zinc is an essential trace element in human biology that is known to be necessary for many biologic functions, such as growth, appetite, testicular maturation, skin integrity, mental activity, wound healing, and immune system maintenance. Approximately 300 enzymes are known to require zinc for their activities. Zinc deficiency in humans is widespread and is more prevalent in areas where the population subsists on cereal proteins. Clinical manifestations of zinc deficiency include: growth retardation, hypogonadism in males, neurosensory disorders, cell-mediated immunological dysfunctions, increased maternal morbidity, premature delivery, and adversely affects the proliferation, regulation and maturity of lymphocytes.
Zinc has been shown to be an essential element for the function of the immune system. Regarding the effect of zinc on allergies, it is known that mast cells have been implicated as mediators of Type I allergic reactions. Mast cell derived reactions result from the release of histamine, heparin, prostaglandins, SRS-A, and various vasoactive amines from granules on the surface of mast cells, possibly including kinins. One product of mast cell-induced inflammation is fever. The inhibitory effect of zinc on histamine release from mast cells are attributed to its action on the stabilization of the mast cell membrane. Zinc ions were found to stabilize cell plasma membranes and prevent induced histamine and vasoactive amine release from tissue mast cells. It has been observed that unsequestered zinc ions (4 to 20 millimolar) are released in inflammation from mast cell granules suggesting a common linkage with inflammation. Zinc is a competitive antagonist of the calcium-dependent IgE and f-met peptide mediated histamine release from human basophils and suggested that zinc compounds might be considered for the treatment of autoimmune disorders.
Zinc compounds are acknowledged as anti-inflammatory agents, as astringents and beneficial in wound healing, and have antimicrobial, antifungal and antiviral activity. Zinc is the active agent in formulations to treat diaper rash, decubitus ulcers, and abrasions. Zinc stabilizes the cell membranes and inhibits the formation of free radicals. Zinc also strengthens the integrity of blood vessel walls by reducing the membrane permeability and stopping bleeding. Unlike other metals, zinc is virtually nontoxic.
Canadian researchers reduced the progression of experimental osteoarthritis in dogs by inhibiting inducible nitric oxide synthase (NOS). Pelletier J P, et al. Arthritis and Rheumatism 1998;41:1275-1286. Pelletier reported that osteoarthritis cartilage produced an increased amount of nitric oxide (NO) due to an increased level of inducible nitric oxide synthase in cartilage chondrocytes. Nitric oxide plays an important role in autoimmunity and inflammation. Normal cartilage does not produce NO or express NOS unless stimulated with cytokines. In the joint, NO, produced in response to cytokine stimulation, exerts a number of catabolic effects on chondrocyte functions which would be expected to promote the degradation of articular cartilage. These effects of NO on chondrocytes include: inhibition of collagen and proteoglycan synthesis, activation of metalloproteinases, increased susceptibility to injury by other oxidants, inhibition of actin polymerization, and apoptosis. NSAIDs, such as aspirin, and to a lesser extent, sodium salicylate, and tetracycline inhibit the expression of NOS protein. Clancy R M, Amin A R, Abramson S B. Arthritis and Rheumatism 1998;41:1141-1151. Zinc is a very potent inhibitor of nitric oxide synthase (NOS). Cuajungco M P, Lees G J Neurobiol Disease 1997;4(3-4):137-69.
The dosage range of an oral zinc compound is 30 to 60 mg per day in divided doses. The suggested topical dosage range is 10 to 20 mg. Zinc compounds are selected from a group consisting of, but not limited to: zinc sulfate, zinc chloride, zinc acetate, zinc phenol sulfonate, zinc borate, zinc bromide, zinc nitrate, zinc glycerophosphate, zinc benzoate, zinc carbonate, zinc citrate, zinc hexafluorosilicate, zinc diacetate trihydrate, zinc oxide, zinc peroxide, zinc salicylate, zinc silicate, zinc stannate, zinc tannate, zinc titanate, zinc tetrafluoroborate, zinc gluconate, and zinc glycinate.
The topical use of zinc compounds is well known in the art. Kelly, U.S. Pat. No. 5,624,675, discloses the use of a zinc salt, such zinc acetate or zinc propionate as a genital lubricant to reduce the risk of HIV infection. The above cited patent does not teach or suggest the use of the method and composition outlined in the present invention.
Aloe vera has been well reported to have anti-inflammatory and analgesic properties, but its use in treating musculoskeletal disorders has only recently been described. One study treated patients with a diagnosis of fibromyalgia and/or chronic fatigue syndrome with aloe vera gel extract and found that there was a remarkable reduction in initial symptom severity and continued improvement during the course of the study. Dykman K D, Tone C, Ford C, Dykman R A, The effects of nutritional supplements on the symptoms of fibromyalgia and chronic fatigue syndrome. Interg Physiol Behav Sci 1998 January-March;33:61-71. Both topical and oral treatments with aloe vera were found to increase the synthesis of glycosaminoglycans and enhance would healing. Chithra P, Sajithlal G B, Chandrakasan G, Influence of Aloe vera on the glycosaminoglycans in the matrix of healing dermal wounds in rats. J Ethnopharmacol 1998 January;59(3). 179-86. Aloe vera also increased the biosynthesis of collagen. Chithra P, Sajithlal G B, Chandrakasan G, Influence of Aloe vera on collagen turnover in healing of dermal wounds in rats. Indian J Exp Biol 1998 Septemeber;36(9):896-901. Aloe vera mixed with a nitric oxide inhibitor (L-NAME) improved wound healing and prevented dermal ischemia by reversing the effects of thromboxane synthase. Effect of the combination of Aloe vera, nitroglycerin, and L-NAME on wound healing in the rat excisional model. J. Altern Complement Med 1997 Summer;3(2): 149-53.
The use of Aloe vera is well known in the art. Carpenter et al, U.S. Pat. No. 5,786,342, discloses a method of reducing symptoms associated with chronic respiratory diseases using acetylated mannan from aloe vera. Strickland et al, U.S. Pat. No. 5,824,659, discloses the use of a oligosaccharide from Aloe to inhibit the loss of skin immunocompetency from ultraviolet irradiation.
Glucosamine from exogenous sources (food and supplements) may stop the progression of cartilage degradation and stimulate the production of new cartilage. Glucosamine absorbed by the gastrointestinal tract undergoes significant first-pass metabolism in the liver, with the resulting 26% bioavailibility. It is incorporated into plasma proteins as a result of hepatic metabolism, and concentrates in the articular cartilage. Clinical improvement of symptoms has been seen as early as one week after oral administration of glucosamine sulfate and has persisted for up to four weeks after discontinuation. Barclay T S, Tsourounis C, McCart G M. Glucosamine. Annals of Pharmacotherapy 1998;32:574-79. In a review of the studies on glucosamine sulfate, the reviewers concluded that the chondroprotective agent reduces joint pain and tenderness, provided pain relief and improved mobility in patients with osteoarthritis. Da Camara C C, Dowles G V, Glucosamine sulfate for osteoarthritis.
Ann Pharmacotherapy 1998 May;32(5):580-7.
Several commercial forms of glucosamine are available, including the sulfate, hydrochloride, and N-acetylglucosamine (NAG). Glucosamine hydrochloride has a higher concentration of glucosamine than the sulfate form. NAG is rapidly metabolized to make proteins and provides less glucosamine for cartilage repair. The composition of the invention could include one or a combination of the glucosamine forms. Patients have reported a more rapid response with higher oral dosages of glucosamine. The dosage range for glucosamine can vary from 500 mg to 3000 mg a day, in divided doses, depending on body weight and severity of symptoms. The suggested topical dosage ranges from 500 to 1,000 mg. Adverse effects reported from glucosamine are gastrointestinal, such as heartburn and epigastric pain. Because the half-life of glucosamine in the blood is relatively short, a topical composition of the compound could avoid the adverse effects and provide a more uniform blood level.
Chondroitin sulfates are the major GAG in cartilage, and has a synergistic effect with glucosamine. Dosage range of chondroitin sulfate is 250 mg to 1,000 mg per day in divided doses. A dosage range for a topical composition could be 100 to 300 mg.
The use of glucosamine and chondroitin as an oral composition are well known in the art. Henderson, U.S. Pat. No. 5,364,845, discloses a composition of glucosamine and chondroitin as an oral composition. Morrison, U.S. Pat No. 3,895,107, discloses the use of chondroitin sulfate, derived from shark cartilage as a treatment for inhibiting the development of atherosclerotic lesions. None of the above cited patents teach or suggest the use of the method or composition outlined in the present invention.
The composition may also include various additional agents and ingredients such as: preservatives, antioxidants, stabilizers, surfactants, emollients, anti-infective agents, adjuvants, thickening and gelling agents, anthocyanidins, proanthocyanidins, amino sugars, glycosaminoglycans, nitric oxide synthase inhibitors, zinc salts, manganese, magnesium, boron, and herbal derivatives.

Method and composition to treat musculoskeletal disorders


A software system provides security against unauthorized operations initiated by software code supplied by an untrusted source. The allowed operations that are associated with the software code are determined. A thinned interface is generated which permits the software code to successfully call only the allowed operations. The software code is independent of a security environment of the system. The thinned interface operates in at least one version of the security environment. The software code and the thinned interface are activated within the system.

The invention is directed to systems that incorporate instructions supplied by external, non-trusted sources and, more particularly, to dynamic, transparent multi-level security for such systems.
Many systems, such as telephone network products and data communication products, include externally developed software applications that require calls to various functions within the system. It is desirable, however, to limit the functions that can be called by the application to those that are necessary or that are approved for the developer of the application. Therefore, the systems typically include a security model that requires the application to include code which requests authorization and receives permission or denial to carry out the respective function call. As an example, the Java environment includes security devices such as a security manager, a byte code verifier and a class loader. However, the security devices of a respective environment may be non-backward compatible with earlier versions. In the Java environment, for example, the security devices in version 1.2 are not backward compatible with those in versions 1.1 and 1.0.2, and the security devices in version 1.1 are not backwards compatible with those in version 1.0.2. Thus, an application program written in a respective version of Java is not compatible with other versions.
Furthermore, in some programming environments, such as in the Java environment, the security devices provide multi-level security but are not transparent, namely the user code must explicitly interact with the system, and the security devices are not dynamic, namely that off-line changes to the system may be necessary. Alternatively, the security devices are code transparent but do not provide multi-level security.
It is therefore desirable to restrict access to a system by untrusted external code using a dynamic, transparent, multi-level security device that is not code dependent or dependent upon the version of the environment used.
The present invention provides a security defense that is transparent to the software application. The code of the application is written at a level above the security devices and is version independent.
The invention includes a security access mediator that receives and authenticates the externally supplied software code, determines the functions that are permitted to the respective application, and then dynamically generates an interface that allows the software application to call only a portion of the full set of function calls. The subset of function calls in the interface is determined as a function of one or more of the following: the credentials of the user that supplies the software application and the user""corresponding service access levels; the service access levels assigned to the software application; the device that will run the software application; the time and circumstances under which the application is to be run; the credentials of the server that supplies the software application and its corresponding service access levels; the functions that are to be called by the application; and when the downloading of the application is requested by another application, the credentials of the requesting application and its corresponding service access levels. Further, the security access mediator is operable in one or more versions of the programming environment.
In accordance with the invention, a method and an apparatus provide security against unauthorized operations in a system that includes software code supplied by an untrusted source. The allowed operations that are associated with the software code are determined. A thinned interface which permits the software code to successfully call only the allowed operations is generated. The software code is independent of a security environment of the system. The thinned interface operates in at least one version of a security environment. The software code and the thinned interface are activated within the system.
Other features and advantages of the present invention will become apparent from the following detailed description of the invention with reference to the accompanying drawings.

Method of and apparatus for providing multi-level security access to system