WO2019116351A2 - Compositions and methods for treating and preventing a staphylococcal infection - Google Patents

Abstract

The present invention relates generally to treatment regimes, compositions, and methods of use to treat and/or prevent a Staphylococcal infection. The compositions can contain compounds found in green tea.

Description

COMPOSITIONS AND METHODS FOR TREATING AND PREVENTING A

STAPHYLOCOCCAL INFECTION

CROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of priority to U.S. Provisional Patent

Application Serial No. 62/598759, filed December 14, 2017, the entire contents of which are hereby incorporated by reference in their entirety.

BACKGROUND OF THE INVENTION

A. Field of the Invention

[0002] The present invention relates to formulations containing a mixture of compounds capable of treating and preventing a Staphylococcal infection.

B. Description of Related Art

[0003] Staphylococcus aureus is one of the most common and detrimental human pathogens. It is estimated that approximately one third of the human population is colonized with S. aureus. S. aureus can cause food poisoning and infect many organs including the skin, joints, bones, lungs, and heart. S. aureus can cause skin infections such as boils, cellulitis, impetigo, and Staphylococcal scalded skin syndrome and can also cause toxic shock syndrome, bacteremia, sepsis, and septic arthritis. S. aureus infections can be life threatening in even otherwise healthy people. (Mayo Foundation for Medical Education and Research (MFMER). 2017)

[0004] Modern pharmacological antibiotics utilized to treat S. aureus have been highly effective for treatment of infections until recently, where overuse and misuse has allowed mechanisms for S. aureus resistance to develop. (Stewart and William Costerton 2001) (Cosgrove et al. 2003.) Resistance to b-lactam antibiotics (e.g. methicillin, oxacillin) is one example. (Tiemersma et al. 2004) (Lee 2003) (Panlilio et al. 1992). b-lactams bind and inhibit the S. aureus penicillin binding proteins (PBPs), DD-transpeptidases, of non-resistant S. aureus. (Fuda et al. 2004) (Brown and Reynolds 1980). Inhibiting DD-transpeptidases blocks completion and repair of the bacterial cell wall and inhibits cellular division. Resistance to b-lactams is caused by a point mutation in PBP (PBP2a) that blocks b-lactam binding b-lactam resistant S. aureus, called methicillin-resistant S. aureus (MRSA), is a rapidly growing and particularly virulent bacterium that is life threatening and an escalating medical concern around the world. (King et al. 2006) (Kallen et al. 2010) (Enright et al. 2002). MRSA infections have become common in hospitals and are increasing in frequency in the community. It is estimated that nearly a million MRSA infections occur in the United States alone each year, resulting in 20,000 to 40,000 deaths. Current therapies are suboptimal in terms of efficacy, safety, side effects, and dosing. (Holland, Arnold, and Fowler 2014) (Kallen et al. 2010) (Tiemersma et al. 2004) (Panlilio et al. 1992) (Rodvold and McConeghy 2014) (Holzgrabe and Abele-Horn 2012) (Enright et al. 2002).

[0005] In an effort to battle these increasingly drug resistant bacteria, a push for development of alternative treatments has been of high importance.

SUMMARY OF THE INVENTION

[0006] The present invention provides a solution to the current problems facing treatment and prevention of Staphylococcal infection. The inventors have surprisingly found that a combination of several compounds found in Camellia sinensis (green tea) can treat and prevent a Staphylococcal infection, including MRSA, without development of resistance. Further, a synergistic antibacterial effect was shown when the combination of compounds disclosed herein is further combined with a b-lactam, such as oxacillin.

[0007] In some aspects, disclosed is a composition containing any one of, any combination of, or all seven biomarkers found in green tea. In some instances the composition includes any one of, any combination of, or all of biomarker 1 having an accurate mass of 188.1020 amu, biomarker 2 having an accurate mass of 126.0306 amu, biomarker 3 having an accurate mass of 152.0233 amu, biomarker 4 having an accurate mass of 180.0641 amu, biomarker 5 having an accurate mass of 290.0790 amu, biomarker 6 having an accurate mass of 306.0739 amu, and biomarker 7 having an accurate mass of 360.1328 amu, wherein each biomarker is found in green tea.

[0008] The amounts of the ingredients within the composition can vary (e.g., amounts can be as low as 0.000001% to as high as 80% w/w or any range therein). In some instances, biomarker 1 has a relative abundance of at least 3.66%, biomarker 2 has a relative abundance of at least 13.52%, biomarker 3 has a relative abundance of at least 2.72%, biomarker 4 has a relative abundance of at least 213.52%, biomarker 5 has a relative abundance of at least 5.84%, biomarker 6 has a relative abundance of at least 2.00%, and biomarker 7 has a relative abundance of at least 18.16%, wherein the relative abundance is relative to 0.01 mg/ml curcumin spiked in 1 mg/ml of the composition. In some instances, biomarker 1 has a relative abundance of at most 5.50%, biomarker 2 has a relative abundance of at most 20.28%, biomarker 3 has a relative abundance of at most 4.08%, biomarker 4 has a relative abundance of at most 320.28%, biomarker 5 has a relative abundance of at most 8.76%, biomarker 6 has a relative abundance of at most 3.00%, and biomarker 7 has a relative abundance of at most 27.24%, wherein the relative abundance is relative to 0.01 mg/ml curcumin spiked in 1 mg/ml of the composition.

[0009] In some instances, the composition includes at least 2, 3, 4, 5, 6, or 7 of biomarkers 1 to 7. In some instances, the mass of each biomarker is the mass as determined by a Direct Analysis in Real Time-TOF (DART-TOF) mass spectrometer.

[0010] In some aspects, at least one of biomarkers 1 through 7 are synthetically obtained. In some aspects, at least one of biomarkers 1 through 7 are obtained from an organism. In some instances, at least one of biomarkers 1 through 7 are obtained from green tea. In another instance, the composition has at least 90%, at least 95%, or at least 98% batch-to-batch chemical consistency of relative abundance for the biomarkers.

[0011] In some instances, the composition further includes an antibiotic. In some instances, the antibiotic is a drug. In some instances, the antibiotic is a b-lactam. In some instances, the composition contains an effective dose of the composition. In some instances, the composition contains an effective dose of the composition and 200 mg or less of a b- lactam. In some instances, the composition contains a synergistic amount of at least one of biomarkers 1 through 7 and an antibiotic. In some instances, the composition contains a concentration of the antibiotic that is less than an effective amount of the antibiotic if administered without a composition containing biomarkers 1 through 7. In some instances, the composition contains 1000 mg or less, 500 mg or less, 300 mg or less, 200 mg or less, 100 mg or less, 50 mg or less, 25 mg or less, 10 mg or less, 9 mg or less, 8 mg or less, 7 mg or less, 6 mg or less, 5 mg or less, 4 mg or less, 3 mg or less, 2 mg or less, or 1 mg or less, etc. of the antibiotic.

[0012] In some instances, the composition comprising one or more of biomarkers 1 to

7 is formulated for oral administration. In some instances, the composition is one or more of a lozenge, a powder, a tablet, a capsule, a delayed release capsule, a quick release capsule, a gel-cap, a gelatin, a liquid solution, and/or a dissolvable film. In some aspects, any of the compositions herein are formulated for topical application. In some instances, the composition is one or more of an emulsion, solution, foam, spreadable solid, soap, serum, etc. In some instances, any of the compositions herein are formulated for intravenous administration and/or intranasal delivery.

[0013] In some instances, the composition is capable of treating and/or preventing a

Staphylococcal infection. In some instances, the composition is capable of bactericidal activity and bacteriostatic activity against Staphylococcus. In some instances, the composition is capable of providing anti-bacterial activity without causing resistance to the composition. In some instances, the composition is capable of providing a synergistic effect with a b-lactam.

[0014] In some instances, the compositions contain a topical, oral, nutraceutical, and/or pharmaceutically acceptable carrier. In some instances, the topical, oral, nutraceutical, and/or pharmaceutically acceptable carrier can be a naturally or a non-naturally occurring compound or structure. In some instances, the composition further contains a pH adjuster, structuring agents, inorganic salt, preservative, stabilizer, bulking agent, and/or excipient. In some instances, the preservative, stabilizer, bulking agent, and/or excipient can be a naturally or a non-naturally occurring compound or composition. In some instances, the composition is a topical composition. In some instances, the composition is suitable for oral administration. In some instances, the composition is a nutraceutical composition. In some instances, the composition is a pharmaceutical composition. In some instances, the composition is coated with a polymer. In some instances, the polymer is a naturally or a non-naturally occurring polymer. In some aspects of the invention, the formulated composition can be comprised in a solid nanoparticle, a lipid-containing nanoparticle, a lipid-based carrier, a sealed conduit, a sealed bag, or any combination thereof. In some instances, the composition can be formulated for administration by injection.

[0015] In some aspects, the composition may further comprise one or more ingredients described herein. For example, the composition may comprise one or more additional ingredients selected from one or more pH adjusters, structuring agents, inorganic salts, preserving agents, fillers, disintegrating agents, wetting agents, emulsifiers, suspending agents, sweeteners, flavorings, fragrance, antibacterial agents, antifungal agents, lubricating agents, vitamins, polymers, siloxane containing compounds, essential oils, and dispensing agents. Each carrier is acceptable in the sense of being compatible with the other ingredients of the formulation and not injurious to the subject. In some aspects, the carrier can include at least one hydrophilic polymeric compound selected from the group consisting of a gum, a cellulose ether, an acrylic resin, a carbohydrate carrier, talc, lactose, mannitol, glucose, water, gelatin, a protein-derived compound, polyvinyl pyrrolidone, magnesium stearate, and any combination thereof. The amounts of such ingredients can range from 0.0001% to 99.9% by weight or volume of the composition, or any integer or range in between as disclosed in other sections of this specification, which are incorporated into this paragraph by reference.

[0016] In some instances, the composition can be stored for one month, 6 months, 12 months, 18 months, 24 months, three years, or more at room temperature.

[0017] In some instances, the composition contains a dose of the biomarkers contained therein. In some instances, a dose is 1 pg to 10 grams of any one or more of the biomarkers. In some instances, the composition contains a sub-dose of one or more of the biomarkers. In some instances, the composition contains 0.1 mg to 3 g, 1 mg to 1 g, 1 mg to 500 mg, 1 to 300 mg, 1 to 100 mg, 1 to 10 mg, 1 mg, 2 mg, 3 mg, or 4 mg of any one or more of the biomarkers, or any range therein.

[0018] In some aspects, the composition contains a second agent capable of treating or preventing a Staphylococcal infection. In some instances, the composition does not comprise a second agent capable of treating or preventing a Staphylococcal infection.

[0019] Also disclosed is a method of treating or preventing a Staphylococcal infection in a subject. The method comprises administering any one or more of the compositions of the present invention to the subject. In some instances, the subject has been diagnosed with a Staphylococcal infection. In some instances, the subject has been diagnosed with a MRSA infection. In some instances, the method is for treating and/or preventing Staphylococcal infection. In some instances, the method is for treating and/or preventing a MRSA infection. In some instances, the method is to decrease the amount of a secondary antibacterial used in the method. In some instances, the second antibacterial is a b-lactam. In some instances, the second antibacterial is oxacillin.

[0020] In some instances, the method includes administering a second antibiotic. The antibiotic can be a b-lactam. The antibiotic can be oxacillin. In some instances, the method includes administering an effective dose of the composition. In some instances, the method includes administering an effective dose of the composition and 200 mg or less of a b-lactam in a 24 hour period. In some instances, the method includes administering a synergistic amount of at least one of biomarkers 1 through 7 and an antibiotic. In some instances, the method includes administering a concentration of the antibiotic that is less than an effective amount of the antibiotic if administered without a composition containing biomarkers 1 through 7. In some instances, the method includes administering a concentration of the composition containing biomarkers 1 through 7 that is less than an effective amount of the composition containing biomarkers 1 through 7 if administered without a composition containing a second antibiotic. In some instances, the method includes administering a concentration of the antibiotic and a concentration of the composition containing biomarkers 1 through 7 that is less than an effective amount of the antibiotic and the composition containing biomarkers 1 through 7 if administered alone. In some instances, the method includes administering 1000 mg or less, 500 mg or less, 300 mg or less, 200 mg or less, 100 mg or less, 50 mg or less, 25 mg or less, 10 mg or less, 9 mg or less, 8 mg or less, 7 mg or less, 6 mg or less, 5 mg or less, 4 mg or less, 3 mg or less, 2 mg or less, or 1 mg or less, etc. of the antibiotic in a 24 hour period.

[0021] In some instances, the subject is administered a total sum of between 1 and

5,000 mg, between 10 and 1,500 mg, between 50 and 1,000 mg, or between 100 and 500 mg of any one or more of the biomarker(s) during a 24 hour period. In some aspects of any of the methods described herein, the subject is administered any one of the compositions disclosed herein at least once a day. In some instances, the subject is administered any one of the compositions disclosed more than once a day. In some instances, the subject is administered any one of the compositions disclosed herein twice a day. In some instances, the composition is administered at least once a day for at least three days. In some aspects, the subject is administered any one of the compositions disclosed herein daily for at least five days. In some aspects, the subject is administered any one of the compositions disclosed herein daily for at least ten days. In some aspects, the subject is administered any one of the compositions disclosed herein daily for at least fourteen days. In some aspects, the subject is administered any one of the compositions disclosed herein daily for at least two weeks, at least a month, at least two months, at least half a year, at least a year, or at least several years. In some aspects, the subject is administered any one of the compositions disclosed herein every other day for at least six days. In some aspects, the subject is administered any one of the compositions disclosed herein every other day for at least ten days. In some aspects, the subject is administered any one of the compositions disclosed herein every other day for at least fourteen days. In some aspects, the subject is administered any one of the compositions disclosed herein every other day for at least two weeks, at least a month, at least two months, at least half a year, at least a year, or at least several years.

[0022] In some aspects, there is disclosed a method of producing any one of the compositions disclosed herein by producing a composition having an at least 90%, at least 95%, or at least 98% batch-to-batch chemical consistency of relative abundance for the biomarkers.

[0023] Kits that include the compositions of the present invention are also contemplated. In some embodiments, the composition is comprised in a container. The container can be a bottle, dispenser, package, or a straw. The container can dispense a predetermined amount of the composition. In certain aspects, the compositions are dispensed as a pill, a tablet, a capsule, a transdermal patch, an edible chew, a cream, a lotion, a gel, spray, mist, dollop, a powder, or a liquid. The container can include indicia on its surface. The indicia can be a word, an abbreviation, a picture, or a symbol.

[0024] It is contemplated that any embodiment discussed in this specification can be implemented with respect to any method or composition of the invention, and vice versa. Furthermore, compositions of the invention can be used to achieve methods of the invention.

[0025] Also contemplated is a product that includes the composition of the present invention. In some aspects, the product can be a nutraceutical product. The nutraceutical product can be those described in other sections of this specification or those known to a person of skill in the art. In some aspects, the product can be a pharmaceutical product. The pharmaceutical and/or nutraceutical product can be those described in other sections of this specification or those known to a person of skill in the art. Non-limiting examples of products include a pill, a tablet, an edible chew, a capsule, a cream, a lotion, a gel, a spray, a mist, a dissolving film, a transdermal patch, or a liquid, etc.

[0026] Also disclosed are the following Embodiments 1 to 36 of the present invention. Embodiment 1 is a composition comprising the following biomarkers: biomarker 1 having an accurate mass of 188.1020 amu and having a relative abundance of at least 3.66%; biomarker 2 having an accurate mass of 126.0306 amu and having a relative abundance of at least 13.52%; biomarker 3 having an accurate mass of 152.0233 amu and having a relative abundance of at least 2.72%; biomarker 4 having an accurate mass of 180.0641 amu and having a relative abundance of at least 213.52%; biomarker 5 having an accurate mass of 290.0790 amu and having a relative abundance of at least 5.84%; biomarker 6 having an accurate mass of 306.0739 amu and having a relative abundance of at least 2.00%; and biomarker 7 having an accurate mass of 360.1328 amu and having a relative abundance of at least 18.16%, wherein the biomarkers are found in green tea; and wherein the relative abundance is relative to 0.01 mg/ml curcumin spiked in 1 mg/ml of the composition. Embodiment 2 is the composition of embodiment 1, wherein the biomarkers contained therein have a relative abundance of at most: biomarker 1 of 5.50%; biomarker 2 of 20.28%; biomarker 3 of 4.08%; biomarker 4 of 320.28%; biomarker 5 of 8.76%; biomarker 6 of 3.00%; and biomarker 7 of 27.24%, wherein the relative abundance is relative to 0.01 mg/ml curcumin spiked in 1 mg/ml of the composition. Embodiment 3 is the composition of any of embodiments 1 to 2, wherein the mass of each biomarker is the mass as determined by a Direct Analysis in Real Time-TOF (DART-TOF) mass spectrometer. Embodiment 4 is the composition of any one of embodiments 1 to 3, wherein at least one of the biomarker(s) are synthetically obtained. Embodiment 5 is the composition of any one of embodiments 1 to 4, wherein at least one of the biomarker(s) are isolated from a plant. Embodiment 6 is the composition of embodiment 5, wherein at least one of the biomarkers(s) are isolated from green tea. Embodiment 7 is the composition of any one of embodiments 1 to 6, wherein the composition has an at least 90%, preferably at least 95%, or at least 98% batch-to-batch chemical consistency of relative abundance for the biomarkers. Embodiment 8 is the composition of any one of embodiments 1 to 7, wherein the composition further comprises a preservative. Embodiment 9 is the composition of any one of embodiments 1 to 8, wherein the composition further comprises an antibiotic. Embodiment 10 is the composition of embodiment 9, wherein the antibiotic is a b-lactam. Embodiment 11 is the composition of embodiment 10, wherein the b-lactam is oxacillin. Embodiment 12 is the composition of any one of embodiments 1 to 11, wherein the composition is formulated for oral administration. Embodiment 13 is the composition of embodiment 12, wherein the composition is in a lozenge, a powder, a tablet, a gel-cap, a gelatin, a liquid solution, a syrup, an oil, and/or a dissolvable film. Embodiment 14 is the composition of any one of embodiments 1 to 11, wherein the composition is formulated for topical administration, administration through injection, and/or intranasal administration. Embodiment 15 is the composition of embodiment 14, wherein the composition is in a liquid solution, emulsion, powder, or patch. Embodiment 16 is the composition of any of embodiments 1 to 15, wherein the composition is formulated to treat and/or prevent a staphylococcal infection. Embodiment 17 is the composition of any of embodiments 1 to 16, wherein the composition is formulated to treat and/or prevent a methicillin resistant Staphylococcus aureus (MRS A). Embodiment 18 is the composition of any of embodiments 1 to 17, wherein the composition comprises at most 200 mg of a b-lactam. Embodiment 19 is a kit comprising: a first composition comprising a green tea extract formulated to be administered to a subject; and a second composition comprising at least one antibiotic formulated to be administered to a subject. Embodiment 20 is the kit of embodiment 19, wherein the green tea extract is an aqueous extract. Embodiment 21 is the kit of embodiment 19, wherein the first composition comprises any one of the compositions of embodiments 1 to 18. Embodiment 22 is the kit of any of embodiments 19 to 21, wherein the at least one antibiotic is a b-lactam. Embodiment 23 is the kit of any of embodiments 19 to 22, wherein the kit further comprises an additional antibiotic. Embodiment 24 is a method of treating a subject at risk for or having a staphylococcal infection, the method comprising administering a composition containing a green tea extract to the subject, wherein at least one symptom of the a staphylococcal infection is ameliorated in the subject and/or the onset of the staphylococcal infection is delayed in comparison to the expected onset of the staphylococcal infection if the patient had not been treated. Embodiment 25 is the method of embodiment 24, wherein the green tea extract is an aqueous extract. Embodiment 26 is the method of embodiment 24, wherein the composition containing a green tea extract comprises the composition of any one of embodiments 1 to 18. Embodiment 27 is the method of any of embodiments 24 to 26, wherein the subject is treated for a methicillin resistant Staphylococcus aureus (MRSA) infection and/or wherein a MRSA infection is prevented. Embodiment 28 is the method of any of embodiments 24 to 27, wherein the subject is diagnosed as having a methicillin resistant Staphylococcus aureus (MRSA) infection. Embodiment 29 is the method of any one of embodiments 24 to 28, wherein the subject is administered a total amount of between 1 and 10,000 mg, between 10 and 5,000 mg, between 50 and 2,500 mg, or between 100 and 1,000 mg of the green tea extract during a 24 hour period. Embodiment 30 is the method of any one of embodiments 24 to 29, wherein the composition containing the green tea extract is administered orally. Embodiment 31 is the method of any one of embodiments 24 to 29, wherein the composition containing the green tea extract is administered topically. Embodiment 32 is the method of any of embodiments 24 to 31, wherein the method further comprises administering at least one antibiotic to the subject. Embodiment 33 is the method of embodiment 32, wherein the at least one antibiotic is a b-lactam. Embodiment 34 is the method of any one of embodiments 32 to 33, wherein a second composition comprises the at least one antibiotic. Embodiment 35 is the method of any of embodiments 33 to 34, wherein the subject is administered 200 mg or less of the b- lactam in a 24 hour period. Embodiment 36 is a method of producing a composition of any of embodiments 1 to 18, wherein the method of producing produces a composition having an at least 90%, preferably at least 95% or at least 98% batch-to-batch chemical consistency of relative abundance for the biomarkers.

[0027] “Therapeutic agent” encompasses the compounds specifically claimed herein.

It also encompasses such compounds together with nutraceutical and/or pharmaceutically acceptable salts thereof. Useful salts are known to those skilled in the art and include salts with inorganic acids, organic acids, inorganic bases, or organic bases. Therapeutic agents useful in the present invention are those compounds that affect a desired, beneficial, and often pharmacological, effect upon administration to a human or an animal, whether alone or in combination with other nutraceutical and/or pharmaceutical excipients or inert ingredients.

[0028] The term“biomarker” refers to the compound defined as the biomarker, analogues thereof, derivatives thereof, or salt forms of any analogue or derivative thereof.

[0029] The term “accurate mass” refers to a measured mass of a molecule experimentally determined for an ion of known charge. The units for accurate mass include atomic mass units (amu) and milli unified atomic mass units (mmu). The term“molecular weight” refers to the average weight of the molecule with all of the different isotopic compositions present in a compound but weighted for their natural abundance.

[0030] The term“relative abundance” refers to the abundance of a compound of interest relative to the abundance of a reference compound. In particular aspects, relative abundance is the raw intensity of a mass spectrometry peak for the compound of interest over the raw intensity of a mass spectrometry peak for a reference compound. In some non limiting instances, the mass spectrometry peaks can be obtained by the use of DART-TOF mass spectrometry. In another particular aspect, the reference compound is a compound that is spiked, or doped, into a sample containing the compound of interest. In yet another particular aspect, the reference compound is a compound that does not exist in the sample previous to its addition to the sample for determining relative abundance. In another particular aspect, the reference compound can be curcumin. [0031] Accurate mass and relative abundances described herein are based on experiments using particular instruments and particular settings and can change from instrument to instrument. There is variability in each measurement. Thus, the accurate mass and relative abundances are defined as being close to as understood by one of ordinary skill in the art. In some non-limiting embodiments the terms are defined to be within 20%, 10%, within 5%, within 1%, or within 0.5%. In some non-limiting embodiments, the accurate mass has an error of within +/- 20 mmu, within 10 mmu, within 5 mmu, or within 1 mmu. In some non-limiting embodiments, the relative abundance has an error of +/- 20%, within 10%, within 5%, within 1%, or within 0.5%.

[0032] The term“substantially” and its variations are defined as being largely but not necessarily wholly what is specified as understood by one of ordinary skill in the art, and in some non-limiting embodiments refers to ranges within 10%, within 5%, within 1%, or within 0.5%.

[0033] “Patient,”“subject,” or“individual” refers to a mammal (e.g., human, primate, dog, cat, bovine, ovine, porcine, equine, mouse, rat, hamster, rabbit, or guinea pig). In particular aspects, the patient, subject, or individual is a human.

[0034] “Inhibiting” or “reducing” or any variation of these terms includes any measurable decrease or complete inhibition to achieve a desired result.

[0035] “Effective” or“effect” or any variation of these terms means adequate to accomplish a desired, expected, or intended result.

[0036] “Treating” or“treat” or any variation of these terms includes any measurable improvement in a disease, condition, or symptom that is being treated or is associated with the disease, condition, or symptom being treated.

[0037] “Preventing” or“prevent” or any variation of these terms means to slow, stop, or reverse progression toward a result. The prevention may be any slowing of the progression toward the result.

[0038] “Analogue” and“analog,” when referring to a compound, refers to a modified compound wherein one or more atoms have been substituted by other atoms, or wherein one or more atoms have been deleted from the compound, or wherein one or more atoms have been added to the compound, or any combination of such modifications. Such addition, deletion or substitution of atoms can take place at any point, or multiple points, along the primary structure comprising the compound.

[0039] “Derivative,” in relation to a parent compound, refers to a chemically modified parent compound or an analogue thereof, wherein at least one substituent is not present in the parent compound or an analogue thereof. A non-limiting example is a parent compound which has been covalently modified. Typical modifications are amides, carbohydrates, alkyl groups, acyl groups, esters, pegylations and the like.

[0040] A“therapeutically equivalent” compound is one that has essentially the same effect in the treatment of a disease or condition as one or more other compounds. A compound that is therapeutically equivalent may or may not be chemically equivalent, bioequivalent, or generically equivalent.

[0041] “Parenteral injection” refers to the administration of small molecule drugs via injection under or through one or more layers of skin or mucus membranes of an animal, such as a human.

[0042] “Bioavailability” refers to the extent to which the therapeutic agent is absorbed from the formulation.

[0043] “Systemic,” with respect to delivery or administration of a therapeutic agent to a subject, indicates that the therapeutic agent is detectable at a biologically significant level in the blood plasma of the subject.

[0044] “Controlled release” refers to the release of the therapeutic agent at such a rate that blood (e.g., plasma) concentrations are maintained within the therapeutic range, but below toxic concentrations over a period of time of about one hour or longer, 6 hours or longer, 12 hours or longer, etc.

[0045] “Pharmaceutically acceptable carrier” refers to a pharmaceutically acceptable solvent, suspending agent or vehicle for delivering a drug compound of the present invention to a mammal such as an animal or human. [0046] “Nutraceutical acceptable carrier” refers to a nutraceutical acceptable solvent, suspending agent or vehicle for delivering a compound of the present invention to a mammal such as an animal or human.

[0047] “Pharmaceutically acceptable” ingredient, excipient or component is one that is suitable for use with humans and/or animals without undue adverse side effects (such as toxicity, irritation and allergic response) commensurate with a reasonable benefit/risk ratio.

[0048] “Nutraceutical acceptable” ingredient, excipient or component is one that is suitable for use with humans and/or animals without undue adverse side effects (such as toxicity, irritation and allergic response) commensurate with a reasonable benefit/risk ratio.

[0049] The term“about” or“approximately” or“substantially unchanged” are defined as being close to as understood by one of ordinary skill in the art, and in some non-limiting embodiments the terms are defined to be within 10%, within 5%, within 1%, or within 0.5%. Further,“substantially non-aqueous” refers to less than 5%, 4%, 3%, 2%, 1%, or less by weight or volume of water.

[0050] The use of the word“a” or“an” when used in conjunction with the term

“comprising” in the claims and/or the specification may mean“one,” but it is also consistent with the meaning of“one or more,”“at least one,” and“one or more than one.”

[0051] As used in this specification and claim(s), the words“comprising” (and any form of comprising, such as“comprise” and“comprises”),“having” (and any form of having, such as“have” and“has”),“including” (and any form of including, such as“includes” and “include”) or“containing” (and any form of containing, such as“contains” and“contain”) are inclusive or open-ended and do not exclude additional, unrecited elements or method steps.

[0052] The compositions and methods for their use can “comprise,” “consist essentially of,” or“consist of’ any of the ingredients or steps disclosed throughout the specification. With respect to the transitional phase“consisting essentially of,” in some non limiting aspects, a basic and novel characteristic of the compositions and methods disclosed in this specification includes the compositions’ abilities to treat and/or prevent a Staphylococcal infection. [0053] Other objects, features and advantages of the present invention will become apparent from the following detailed description. It should be understood, however, that the detailed description and the examples, while indicating specific embodiments of the invention, are given by way of illustration only. Additionally, it is contemplated that changes and modifications within the spirit and scope of the invention will become apparent to those skilled in the art from this detailed description.

BRIEF DESCRIPTION OF THE DRAWINGS

[0054] The following drawings form part of the present specification and are included to further demonstrate certain aspects of the present invention. The invention may be better understood by reference to one or more of these drawings in combination with the detailed description of specific embodiments presented herein.

[0055] FIG. 1 HPLC analysis of 6 different batches of the composition of Example 1 demonstrate API dose consistent quality standards.

[0056] FIG. 2 MRSA growth curves for MRSA untreated (diamonds), treated with clinical doses of oxacillin (squares) and treated with a composition of Example 1 (triangles).

[0057] FIG. 3a and 3b (a) Inhibition of MRSA (top) and S. aureus (bottom) growth on agar treated with 1.0 mg/mL composition of Example 1. No growth was observed for MRSA (top) or S. aureus (bottom) after l8-24h. (b) Chloramphenicol (top) and methicillin (bottom) controls for MRSA and S. aureus respectively.

[0058] FIG. 4a and 4b (a) Oxacillin and (b) composition of Example 1 MIC determination.

[0059] FIG. 5 The composition of Example 1 and oxacillin synergy.

[0060] FIG. 6 MRSA generational studies using the composition of Example 1.

[0061] FIG. 7 MRSA generational studies using synergy concentrations.

[0062] FIG. 8 Human serum suppresses anti-MRSA activity equivalently across treatment regimens. DETAILED DESCRIPTION

[0063] The present invention provides a solution to the current problems facing treatment and prevention of Staphylococcal infection. The inventors have surprisingly found that a combination of several compounds found in Camellia sinensis (green tea) can treat and prevent Staphylococcal infection. Specifically, it is shown herein that a composition containing one or more of biomarkers 1 through 7 is capable of treating and preventing a Staphylococcal infection, including MRSA, without development of resistance. Further, the composition works synergistically when combined with a b-lactam, such as oxacillin.

[0064] Further, it is expected that the composition will have synergistic effect with other antibiotics to treat and/or prevent a Staphylococcal infection. It is also believed that the compounds and compositions disclosed herein are capable of treating and preventing the symptoms associated with a Staphylococcal infection.

A. Active Compounds

[0065] Camellia sinensis (green tea) is one of the most popular beverages worldwide and is known for its beneficial effects on health and disease, with numerous studies that support its ability to act as a powerful anti-oxidant. (Wierzejska 2014) (Yiannakopoulou 2013) (Hayat et al. 2015) (Petrosino and Serafini 2014) (Shirakami, Shimizu, and Moriwaki 2012) (Cao, Shi, and Chen 2013) (Vance et al. 2013). Green tea extracts contain compounds called catechins and polyphenols that are known to be effective scavengers of reactive oxygen species (ROS). (Kim, Quon, and Kim 2014) (Tenore et al. 2015) (Afzal, Safer, and Menon 2015) (Lecumberri et al. 2013) (Yu et al. 2014) (Bhardwaj and Khanna 2013) (Roh et al. 2017) (Braicu et al. 2013). The accumulation of ROS in the body causes a phenomenon called oxidative stress that is implicated in the development of chronic and degenerative illness and in the development of osteoarthritis. (Darvesh and Bishayee 2013)

[0066] The compositions disclosed herein are effective in treating and preventing a

Staphylococcal infection. Specifically, it has now been shown that a composition containing compounds found in green tea can be used to treat and prevent a Staphylococcal infection without inducing resistance. (Radji et al. 2013) (Steinmann et al. 2013) (Novy et al. 2013) (Stevens et al. 2015) (Lau and Plotkin 2013). Further, the composition acts synergistically when combined with a b-lactam, such as oxacillin. [0067] In some aspects, the composition of the present invention can include one or more of the biomarkers found in green tea defined by accurate mass of 188.1020 amu, 126.0306 amu, 152.0233 amu, 180.0641 amu, 290.0790 amu, 306.0739 amu, and 360.1328 amu, and combinations thereof.

[0068] In some embodiments, the biomarker or combination of biomarkers has a 90% batch-to-batch chemical consistency of relative abundance for the biomarkers. In some embodiments, the compound or combination of compounds has a 95% and/or 98% batch-to- batch chemical consistency of relative abundance for the biomarkers.

[0069] In some aspects of the invention, the compounds and derivatives and analogues can be made through known synthetic methods. In some aspects of the invention, the compounds can be synthetically obtained by producing the compound according to methods known to one of skill in the art in chemical synthesis. In some instances, the compound is synthesized through organic chemistry methods.

[0070] In some aspects of the invention, the compounds can be isolated from extracts of an organism such as fruits, plants, animals, fungi, bacteria, and/or archaea. Non-limiting examples of suitable plants include green tea. The compound(s) can be extracted from the organism using known extraction methods, such as contacting the extract with CO2, contacting the extract with H2O, contacting the extract with EtOH, or any combination of EtOfEtbO, and/or with any method utilizing a suitable separation media for purification of the extract. Non-limiting examples of separation medias that can be used for purification include ADS (Nankai University, China), Sephadex (GE Healthcare Life Sciences), and AMBERLITE® (The Dow Chemical Company), etc.

[0071] The extract can include any one of or combination of compounds defined by accurate mass of 188.1020 amu, 126.0306 amu, 152.0233 amu, 180.0641 amu, 290.0790 amu, 306.0739 amu, and/or 360.1328 amu, found in green tea, and any combination thereof.

[0072] In some aspects of the invention, one or more of the compounds of the composition and derivatives and analogues thereof can be made through known synthetic methods known by one of skill in the art and one or more of the compounds of the composition and derivatives and analogues thereof may be isolated from other sources, such as, but not limited to, extracts of fruits and plants. B. Compounds Defined by DART TOF/MS

[0073] The composition of the present invention can include one or more of the compounds defined by accurate mass of about 188.1020 amu, 126.0306 amu, 152.0233 amu, 180.0641 amu, 290.0790 amu, 306.0739 amu, and/or 360.1328 amu, found in green tea and any combination thereof.

[0074] The accurate mass and relative abundances described herein are based on experiments using particular instruments and particular settings and can change from instrument to instrument. There is variability in each measurement. Thus, the accurate mass and relative abundances are defined as being close to as understood by one of ordinary skill in the art. In some non-limiting embodiments the terms are defined to be within 20%, with 10%, within 5%, within 1%, or within 0.5%. In some non-limiting embodiments, the accurate mass has an error of within +/- 20 mmu, within 10 mmu, within 5 mmu, or within 1 mmu. In some non-limiting embodiments, the relative abundance has an error of +/- 20%, within 10%, within 5%, and within 1%, and within 0.5%.

[0075] In some non-limiting examples, the compounds of the present invention can be identified using Direct Analysis in Real Time (DART) Time of Flight/Mass Spectrometry (TOF/MS). Specifically, a JEOL DART™ AccuTOF-mass spectrometer from Jeol USA of Peabody, MA (JMS-T100LC) can be used. Accurate mass can be determined by subtracting the mass of a proton (1.007825 amu) from the measured mass of the ions produced from the sample. The mass of compounds may be determined in a sample by directly introducing the sample to the ion stream by means of a Dip-IT sampler and a Dip-IT sampler holder (IONSENSE™). While no sample preparation is required for a simple analysis with the DART, a chemical doped/spiked solution can be used for quantitation relative to a known quantity. As a non-limiting example, curcumin is not present in green tea extract and can therefore be used to create a quantitative chemical profile of the bioactive molecules. The settings for the DART ion source can be the following:

Gas: He

Flow: 2.52 LPM @ 50 PSI

Temperature: 250 C

Needle Voltage: 3000V

Grid Electrode Voltage: 250V Discharge Electrode Voltage: 400V

The settings for the JEOL AccuTOF MS can be the following:

Peaks Voltage: 1000V

Orifice 1 Temperature: 120 C

Detector Voltage: 2600V

Reflectron Voltage: 990.0V

[0076] Samples can be analyzed in six replicates by DART-TOF MS. These six replicates can be analyzed to create a single, averaged, filtered, and statistically significant DART fingerprint of the sample. This processed fingerprint can then be used to determine the presence of the bioactive markers by comparison of masses. Due to the initial discovery and identification of these bioactive markers, a simple mass comparison is sufficient to determine their presence in any extract or mixture of chemicals.

[0077] All MS have a mass tolerance - a range of acceptable reported masses surrounding the predicted [M+H] or [M-H] value. For the AccuTOF, that mass tolerance is less than 20 millimass units (mmu) (predicted mass +/-10 mmu). Given the same sample and ion source, other TOF-MS may have a higher or lower mass tolerance.

[0078] In some non-limiting examples, the compounds of the present invention can be determined by DART TOF/MS by using a JEOF DART™ AccuTOF-mass spectrometer from Jeol USA of Peabody, MA (JMS-T100FC) executed in the positive ion mode ([M+H]⁺) using the following settings for the DART ion source:

Gas: He

Flow: 3.98 F/min

Needle voltage: 3500 V

Temperature: 300 °C

Electrode 1 Voltage: 150 V

Electrode 2 Voltage: 250 V,

The settings for the JEOF AccuTOF MS can be the following:

Peaks Voltage: 1000V

Orifice 1 Voltage: 20 V Ring Lens Voltage: 5 V

Orifice 2 Voltage: 5 V

Detector Voltage: 2550V

[0079] Calibrations can be performed internally with each sample using a 10%

(weight/volume) solution of PEG 600 from Ultra Chemical of North Kingston, RI that provided mass markers throughout the required mass range of 100-1000 amu. Calibration tolerances can be held to 5 mmu. Samples can be introduced into the DART He plasma using the closed end of a borosilicate glass melting point capillary tube until a signal is achieved in the total-ion chromatogram (TIC). The next sample can then be introduced when the TIC returns baseline levels.

C. Additional Antibiotics

[0080] In some aspects, a second antibiotic and/or therapy can be combined with the compounds of the compositions disclosed herein. In some instances, the agent is a drug. In some embodiments, the compositions disclosed herein further includes at least one additional antibiotic.

[0081] In some instances, the antibiotic is a b-lactam. In some instances the antibacterial is oxacillin. In some instances, the antibiotic is a cephalosporins, nafcillin, a sulfa drug, vancomycin, penicillin, a penicillin derivative, flucloxacillin, dicloxacillin, a cephalosporin, cefazolin, cephalothin, cephalexin, a monobactam, a carbapenem, methicillin, clindamycin, lincomycin, erythromycin, teicoplanin, rifampicin, fusidic acid, a lincosamide, clindamycin, lincomycin, cotrimoxazole, linezolid, daptomycin, ceffaroline fosamiland, televancin, dalbavancin, tedizolid, quinupristin/dalfopristin, etc., or a combination thereof.

D. Amounts of Ingredients

[0082] It is contemplated that the compositions of the present invention can include any amount of the ingredients discussed in this specification. The compositions can also include any number of combinations of additional ingredients described throughout this specification ( e.g ., stabilizers, fillers, pharmaceutically and/or nutraceutical acceptable salts, and/or additional pharmaceutical and/or nutraceutical ingredients). The concentrations of the any ingredient within the compositions can vary. In non-limiting embodiments, for example, the compositions can comprise, consisting essentially of, or consist of, in their final form, for example, at least about 0.0001%, 0.0002%, 0.0003%, 0.0004%, 0.0005%, 0.0006%, 0.0007%, 0.0008%, 0.0009%, 0.0010%, 0.0011%, 0.0012%, 0.0013%, 0.0014%, 0.0015%, 0.0016%, 0.0017%, 0.0018%, 0.0019%, 0.0020%, 0.0021%, 0.0022%, 0.0023%, 0.0024%, 0.0025%, 0.0026%, 0.0027%, 0.0028%, 0.0029%, 0.0030%, 0.0031%, 0.0032%, 0.0033%, 0.0034%, 0.0035%, 0.0036%, 0.0037%, 0.0038%, 0.0039%, 0.0040%, 0.0041%, 0.0042%, 0.0043%, 0.0044%, 0.0045%, 0.0046%, 0.0047%, 0.0048%, 0.0049%, 0.0050%, 0.0051%, 0.0052%, 0.0053%, 0.0054%, 0.0055%, 0.0056%, 0.0057%, 0.0058%, 0.0059%, 0.0060%, 0.0061%, 0.0062%, 0.0063%, 0.0064%, 0.0065%, 0.0066%, 0.0067%, 0.0068%, 0.0069%, 0.0070%, 0.0071%, 0.0072%, 0.0073%, 0.0074%, 0.0075%, 0.0076%, 0.0077%, 0.0078%, 0.0079%, 0.0080%, 0.0081%, 0.0082%, 0.0083%, 0.0084%, 0.0085%, 0.0086%, 0.0087%, 0.0088%, 0.0089%, 0.0090%, 0.0091%, 0.0092%, 0.0093%, 0.0094%, 0.0095%, 0.0096%, 0.0097%, 0.0098%, 0.0099%, 0.0100%, 0.0200%, 0.0250%, 0.0275%, 0.0300%, 0.0325%, 0.0350%, 0.0375%, 0.0400%, 0.0425%, 0.0450%, 0.0475%, 0.0500%, 0.0525%, 0.0550%, 0.0575%, 0.0600%, 0.0625%, 0.0650%, 0.0675%, 0.0700%, 0.0725%, 0.0750%, 0.0775%, 0.0800%, 0.0825%, 0.0850%, 0.0875%, 0.0900%, 0.0925%, 0.0950%, 0.0975%, 0.1000%, 0.1250%, 0.1500%, 0.1750%, 0.2000%, 0.2250%, 0.2500%, 0.2750%, 0.3000%, 0.3250%, 0.3500%, 0.3750%, 0.4000%, 0.4250%, 0.4500%, 0.4750%, 0.5000%, 0.5250%, 0.0550%, 0.5750%, 0.6000%, 0.6250%, 0.6500%, 0.6750%, 0.7000%, 0.7250%, 0.7500%, 0.7750%, 0.8000%, 0.8250%, 0.8500%, 0.8750%, 0.9000%, 0.9250%, 0.9500%, 0.9750%, 1.0%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2.0%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%,

2.6%, 2.7%, 2.8%, 2.9%, 3.0%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3.7%, 3.8%, 3.9%,

4.0%, 4.1%, 4.2%, 4.3%, 4.4%, 4.5%, 4.6%, 4.7%, 4.8%, 4.9%, 5.0%, 5.1%, 5.2%, 5.3%,

5.4%, 5.5%, 5.6%, 5.7%, 5.8%, 5.9%, 6.0%, 6.1%, 6.2%, 6.3%, 6.4%, 6.5%, 6.6%, 6.7%,

6.8%, 6.9%, 7.0%, 7.1%, 7.2%, 7.3%, 7.4%, 7.5%, 7.6%, 7.7%, 7.8%, 7.9%, 8.0%, 8.1%,

8.2%, 8.3%, 8.4%, 8.5%, 8.6%, 8.7%, 8.8%, 8.9%, 9.0%, 9.1%, 9.2%, 9.3%, 9.4%, 9.5%,

9.6%, 9.7%, 9.8%, 9.9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 35%, 40%, 45%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% or any range derivable therein, of at least one of the ingredients that are mentioned throughout the specification and claims. In non-limiting aspects, the percentage can be calculated by weight or volume of the total composition or relative abundance. A person of ordinary skill in the art would understand that the concentrations can vary depending on the addition, substitution, and/or subtraction of ingredients in a given composition. E. Additional Components

[0083] The compound of the present invention can be formulated into any suitable composition form for administration to a human or non-human animal patient.

[0084] The composition may consist of the claimed compound or compounds alone or may include the compound or compounds and any suitable additional component, such as one or more pharmaceutically and/or nutraceutical acceptable carriers, diluents, adjuvants, excipients, or vehicles, such as preserving agents, fillers, disintegrating agents, wetting agents, emulsifying agents, suspending agents, sweetening agents, flavoring agents, perfuming agents, antibacterial agents, antifungal agents, lubricating agents and dispensing agents, depending on the nature of the mode of administration and dosage forms. Each carrier must be acceptable in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient.

1. Excipients

[0085] Excipients employed in the compositions of the present invention can be solids, semi-solids, liquids or combinations thereof. In some instances, the excipients are solids. Compositions of the invention containing excipients can be prepared by any known technique that comprises, for example, admixing an excipient with the claimed compounds. A pharmaceutical composition of the invention contains a desired amount of the claimed compounds per dose unit and, if intended for oral administration, can be in the form, for example, of a tablet, a caplet, a pill, a hard or soft capsule, a lozenge, a cachet, a dispensable powder, granules, a suspension, an elixir, a dispersion, or any other form reasonably adapted for such administration. If intended for topical application, it can be in the form of an emulsion, solution, powder, patch, spreadable solid, etc. If intended for parenteral administration, it can be in the form, for example, of a suspension or transdermal patch. If intended for rectal administration, it can be in the form, for example, of a suppository. Presently preferred are oral dosage forms that are discrete dose units each containing a predetermined amount of the claimed compounds such as tablets or capsules and topical dosage forms.

2. Carriers / Diluents [0086] Suitable carriers or diluents illustratively include, but are not limited to, either individually or in combination, lactose, including anhydrous lactose and lactose monohydrate; starches, including directly compressible starch and hydrolyzed starches (e.g., CELUTAB™ and EMDEX™), mannitol, sorbitol, xylitol, dextrose (e.g., CERELOSE™ 2000) and dextrose monohydrate, dibasic calcium phosphate dihydrate, sucrose-based diluents, confectioner's sugar, monobasic calcium sulfate monohydrate, calcium sulfate dihydrate, granular calcium lactate trihydrate, dextrates, inositol, hydrolyzed cereal solids, amylose, celluloses including microcrystalline cellulose, food grade sources of alpha- and amorphous cellulose (e.g., RexcelJ), powdered cellulose, hydroxypropylcellulose (HPC) and hydroxypropylmethylcellulose (HPMC), calcium carbonate, glycine, clay, bentonite, block co-polymers, polyvinylpyrrolidone, and the like. Such carriers or diluents, if present, constitute in total about 5% to about 99.999%, about 10% to about 85%, and 20% to about 80%, of the total weight of the composition. The carrier, carriers, diluent, or diluents selected preferably exhibit suitable flow properties and, where tablets are desired, compressibility.

3. Disintegrant

[0087] Compositions of the invention optionally can include one or more pharmaceutically and/or nutraceutical acceptable disintegrants as excipients. Suitable disintegrants include, but are not limited to, either individually or in combination, starches, including sodium starch glycolate and pregelatinized com starches, clays, celluloses such as purified cellulose, microcrystalline cellulose, methylcellulose, carboxymethylcellulose and sodium carboxymethylcellulose, croscarmellose sodium, alginates, crospovidone, and gums such as agar, guar, locust bean, karaya, pectin and tragacanth gums. Disintegrants may be added at any suitable step during the preparation of the composition, particularly prior to granulation or during a lubrication step prior to compression. In some instances, such disintegrants, if present, constitute in total about 0.2% to about 30%, about 0.2% to about 10%, or about 0.2% to about 5%, of the total weight of the composition.

4. Binders

[0088] The compositions of the present invention can include binding agents or adhesives particularly for tablet formulations. Such binding agents and adhesives preferably impart sufficient cohesion to the powder being tableted to allow for normal processing operations such as sizing, lubrication, compression and packaging, but still allow the tablet to disintegrate and the composition to be absorbed upon ingestion. Such binding agents may also prevent or inhibit crystallization or recrystallization of a co-crystal of the present invention once the salt has been dissolved in a solution. Suitable binding agents and adhesives include, but are not limited to, either individually or in combination, acacia; tragacanth, sucrose, gelatin, glucose, starches such as, but not limited to, pregelatinized starches, celluloses such as, but not limited to, methylcellulose and carmellose sodium, alginic acid and salts of alginic acid; magnesium aluminum silicate, PEG, guar gum, polysaccharide acids, bentonites, povidone, polymethacrylates, HPMC, hydroxypropylcellulose, and ethylcellulose. In some instances, such binding agents and/or adhesives, if present, constitute in total about 0.5% to about 25%, about 0.75% to about 15%, or about 1% to about 10%, of the total weight of the pharmaceutical composition. Many of the binding agents are polymers comprising amide, ester, ether, alcohol or ketone groups and, as such, can be included in pharmaceutical compositions of the present invention. Polyvinylpyrrolidones is an non-limiting example of a binder used for slow release tablets. Polymeric binding agents can have varying molecular weight, degrees of crosslinking, and grades of polymer. Polymeric binding agents can also be copolymers, such as block co polymers that contain mixtures of ethylene oxide and propylene oxide units. Variation in these units' ratios in a given polymer affects properties and performance.

5. Wetting Agents

[0089] Wetting agents can be used in the compositions of the present invention.

Wetting agent can be selected to maintain the crystal in close association with water, a condition that may improve bioavailability of the composition. Such wetting agents can also be useful in solubilizing or increasing the solubility of crystals. Surfactants can be used as wetting agents. Non- limiting examples of surfactants that can be used as wetting agents in compositions of the invention include quaternary ammonium compounds, for example benzalkonium chloride, benzethonium chloride and cetylpyridinium chloride, dioctyl sodium sulfosuccinate, polyoxyethylene alkylphenyl ethers, poloxamers (polyoxyethylene and polyoxypropylene block copolymers), polyoxyethylene fatty acid glycerides and oils, for example polyoxyethylene (8) caprylic/capric mono- and diglycerides, polyoxyethylene (35) castor oil and polyoxyethylene (40) hydrogenated castor oil, polyoxyethylene alkyl ethers, for example polyoxyethylene (20) cetostearyl ether, polyoxyethylene fatty acid esters, for example polyoxyethylene (40) stearate, polyoxyethylene sorbitan esters, for example polysorbate 20 and polysorbate 80, propylene glycol fatty acid esters, for example propylene glycol laurate, sodium lauryl sulfate, fatty acids and salts thereof, for example oleic acid, sodium oleate and triethanolamine oleate, glyceryl fatty acid esters, for example glyceryl monostearate, sorbitan esters, for example sorbitan monolaurate, sorbitan monooleate, sorbitan monopalmitate and sorbitan monostearate, tyloxapol, and mixtures thereof. In some instances, such wetting agents, if present, constitute in total about 0.25% to about 15%, about 0.4% to about 10%, or about 0.5% to about 5%, of the total weight of the pharmaceutical and/or nutraceutical composition.

6. Lubricants

[0090] Lubricants can be included in the compositions of the present invention.

Suitable lubricants include, but are not limited to, either individually or in combination, glyceryl behenate, stearic acid and salts thereof, including magnesium, calcium and sodium stearates; hydrogenated vegetable oils, colloidal silica, talc, waxes, boric acid, sodium benzoate, sodium acetate, sodium fumarate, sodium chloride, DL-leucine, PEG (e.g., CARBOWAX™ 4000 and CARBOWAX™ 6000 of the Dow Chemical Company), sodium oleate, sodium lauryl sulfate, and magnesium lauryl sulfate. In some instances, such lubricants, if present, constitute in total about 0.1% to about 10%, about 0.2% to about 8%, or about 0.25% to about 5%, of the total weight of the composition.

7. Other Agents

[0091] Surfactant, emulsifier, or effervescent agents can be used in the compositions.

Emulsifying agents can be used to help solubilize the ingredients within a soft gelatin capsule. Non-limiting examples of the surfactant, emulsifier, or effervescent agent include D-sorbitol, ethanol, carrageenan, carboxyvinyl polymer, carmellose sodium, guar gum, glycerol, glycerol fatty acid ester, cholesterol, white beeswax, dioctyl sodium sulfosuccinate, sucrose fatty acid ester, stearyl alcohol, stearic acid, polyoxyl 40 stearate, sorbitan sesquioleate, cetanol, gelatin, sorbitan fatty acid ester, talc, sorbitan trioleate, paraffin, potato starch, hydroxypropyl cellulose, propylene glycol, propylene glycol fatty acid ester, pectin, polyoxyethylene (105) polyoxypropylene (5) glycol, polyoxyethylene (160) polyoxypropylene (30) glycol, polyoxyethylene hydrogenated castor oil, polyoxyethylene hydrogenated castor oil 40, polyoxyethylene hydrogenated castor oil 60, polyoxyl 35 castor oil, polysorbate 20, polysorbate 60, polysorbate 80, macrogol 400, octyldodecyl myristate, methyl cellulose, sorbitan monooleate, glycerol monostearate, sorbitan monopalmitate, sorbitan monolaurate, lauryl dimethylamine oxide solution, sodium lauryl sulfate, lauromacrogol, dry sodium carbonate, tartaric acid, sodium hydroxide, purified soybean lecithin, soybean lecithin, potassium carbonate, sodium hydrogen carbonate, medium-chain triglyceride, citric anhydride, cotton seed oil-soybean oil mixture, and liquid paraffin.

F. Vehicles

[0092] Various delivery systems are known in the art and can be used to administer a therapeutic agent or composition of the invention, e.g., encapsulation in liposomes, microparticles, microcapsules, receptor-mediated endocytosis and the like. Methods of administration include, but are not limited to, parenteral, intra-arterial, intramuscular, intravenous, intranasal, and oral routes. The compositions can be provided in the form of tablets, lozenges, granules, capsules, pills, ampoule, suppositories or aerosol form. The compositions can also be provided in the form of suspensions, solutions, and emulsions of the active ingredient in aqueous or non-aqueous diluents, syrups, granulates or powders.

G. Formulation and Administration

[0093] The composition may, for example, be a pharmaceutical composition

(medicament), and over the counter composition (OTC), a nutraceutical, etc. Compositions according to the present invention include formulations suitable for oral or parenteral routes. Non-limiting examples of specific routes include intradermal, subcutaneous, intramuscular, intravenous, local injection, rectal, intranasal inhalation, insufflation, topical (including transdermal, buccal and sublingual), vaginal, parenteral (including subcutaneous, intramuscular, intravenous and intradermal) and pulmonary administration. The formulations can conveniently be presented in unit dosage form and can be prepared by any methods well known in the art. Such methods include the step of bringing into association the active ingredient (or ingredients) with the carrier, which constitutes one or more accessory ingredients. In general, the formulations are prepared by uniformly and intimately bringing into association the active ingredient with a suitable carrier, such as liquid carriers or finely divided solid carriers or both, and then if necessary shaping the product. Formulations of the subject invention suitable for oral administration can be presented as discrete units such as capsules, cachets or tablets, each containing a predetermined amount of the active ingredient, or as an oil-in-water liquid emulsion, water-in-oil liquid emulsion, or as a supplement within an aqueous solution, for example, a tea. The active ingredient can also be presented as bolus, electuary, or paste. Useful injectable preparations include sterile suspensions, solutions or emulsions of the compound compositions in aqueous or oily vehicles. The compositions can also contain formulating agents, such as suspending, stabilizing and/or dispersing agent. The formulations for injection can be presented in unit dosage form, e.g., in ampoules or in multidose containers, and can contain added preservatives. Alternatively, the injectable formulation can be provided in powder form for reconstitution with a suitable vehicle, including but not limited to sterile pyrogen free water, buffer, dextrose solution, etc., before use. To this end, the compound compositions can be dried by any art-known technique, such as lyophilization, and reconstituted prior to use.

[0094] Formulations suitable for topical administration in the mouth include lozenges comprising the active ingredient in a flavored basis, usually sucrose and acacia or tragacanth, pastilles that include the active ingredient in an inert basis such as gelatin and glycerin, or sucrose and acacia, mouthwashes that include the active ingredient in a suitable liquid carrier, and chocolate comprising the active ingredients.

[0095] Formulations suitable for topical administration according to the subject invention can be formulated as an ointment, cream, suspension, lotion, powder, solution, paste, gel, spray, aerosol or oil. Alternatively, a formulation can comprise a patch or a dressing such as a bandage or adhesive plaster impregnated with active ingredients, and optionally one or more excipients or diluents. In some embodiments, topical formulations preferably comprise compounds that facilitate absorption of the active ingredients through the skin and into the bloodstream.

[0096] Formulations suitable for intranasal administration, wherein the carrier is a solid, include a coarse powder having a particle size, for example, in the range of about 20 to about 500 microns, which is administered in the manner in which snuff is taken, i.e., by rapid inhalation through the nasal passage from a container of the powder held close up to the nose. Suitable formulations wherein the carrier is a liquid for intranasal administration, such as by the non-limiting examples of a nebulizer, include aqueous or oily solutions of the agent. Formulations preferably can include compounds that facilitate absorption of the active ingredients through the skin and into the bloodstream. [0097] Formulations suitable for parenteral administration include aqueous and non- aqueous isotonic sterile injection solutions which can contain antioxidants, buffers, bacteriostats and solutes which render the formulation isotonic with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions which can include suspending agents and thickening agents, and liposomes or other microparticulate systems which are designed to target the compound to blood components or one or more organs. The formulations can be presented in unit-dose or multi-dose or multi-dose sealed containers, such as for example, ampoules and vials, and can be stored in a freeze-dried (lyophilized) condition requiring only the addition of the sterile liquid carrier, for example, water for injections, immediately prior to use. Extemporaneous injection solutions and suspensions can be prepared from sterile powders, granules and tablets of the kind previously described.

[0098] Liquid preparations for oral administration can take the form of, for example, elixirs, solutions, syrups or suspensions, or they can be presented as a dry product for constitution with water or other suitable vehicle before use. Such liquid preparations can be prepared by conventional means with pharmaceutically and/or nutraceutical acceptable additives such as suspending agents (e.g., sorbitol syrup, cellulose derivatives or hydrogenated edible fats); emulsifying agents (e.g., lecithin or acacia); non aqueous vehicles (e.g., almond oil, oily esters, ethyl alcohol, or fractionated vegetable oils); and preservatives (e.g., methyl or propyl p hydroxybenzoates or sorbic acid). The preparations can also contain buffer salts, preservatives, flavoring, coloring and sweetening agents as appropriate.

[0099] For buccal administration, the compositions can take the form of the non limiting examples of tablets or lozenges formulated in a conventional manner.

[00100] For rectal and vaginal routes of administration, the compound compositions can be formulated as solutions (for retention enemas) suppositories or ointments containing conventional suppository bases such as cocoa butter or other glycerides.

[00101] For nasal administration or administration by inhalation or insufflation, the compound compositions can be conveniently delivered in the form of an aerosol spray from pressurized packs or a nebulizer with the use of a suitable propellant, e.g., dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, fluorocarbons, carbon dioxide or other suitable gas. In the case of a pressurized aerosol, the dosage unit can be determined by providing a valve to deliver a metered amount. Capsules and cartridges for use in an inhaler or insufflator (for example capsules and cartridges comprised of gelatin) can be formulated containing a powder mix of the compound and a suitable powder base such as lactose or starch.

[00102] For prolonged delivery, the compound compositions can be formulated as a depot preparation for administration by implantation or intramuscular injection. The compound compositions can be formulated with suitable polymeric or hydrophobic materials (e.g., as an emulsion in an acceptable oil) or ion exchange resins, or as sparingly soluble derivatives, e.g., as a sparingly soluble salt. Alternatively, transdermal delivery systems manufactured as an adhesive disc or patch, which slowly releases the compound compositions for percutaneous absorption, can be used. To this end, permeation enhancers can be used to facilitate transdermal penetration of the compound compositions. Suitable transdermal patches are described in for example, U.S. Pat. No. 5,407,713; U.S. Pat. No. 5,352,456; U.S. Pat. No. 5,332,213; U.S. Pat. No. 5,336,168; U.S. Pat. No. 5,290,561; U.S. Pat. No. 5,254,346; U.S. Pat. No. 5,164,189; U.S. Pat. No. 5,163,899; U.S. Pat. No. 5,088,977; U.S. Pat. No. 5,087,240; U.S. Pat. No. 5,008,110; and U.S. Pat. No. 4,921,475.

[00103] Alternatively, other delivery systems can be employed. Liposomes and emulsions are well-known examples of delivery vehicles that can be used to deliver the compound compositions. Certain organic solvents such as dimethylsulfoxide (DMSO) can also be employed, although usually at the cost of greater toxicity.

[00104] It should be understood that in addition to the ingredients particularly mentioned above, the formulations useful in the present invention can include other agents conventional in the art regarding the type of formulation in question. For example, formulations suitable for oral administration can include such further agents as sweeteners, thickeners, and flavoring agents. It also is intended that the agents, compositions, and methods of this invention be combined with other suitable compositions and therapies.

[00105] In one embodiment, the pharmaceutical and/or nutraceutical compositions of the invention can be administered locally to the area in need of treatment; such local administration can be achieved, for example, by local infusion, by injection, or by means of a catheter. In another embodiment, a compound or composition of the invention is administered in a manner so as to achieve peak concentrations of the active compound at sites of the disease. Peak concentrations at disease sites can be achieved, for example, by intravenously injecting of the agent, optionally in saline, or orally administering, for example, a tablet, capsule or syrup containing the active ingredient.

1. Other Pharmaceutical and/or Nutraceutical Agents

[00106] Pharmaceutical, over the counter (OTC), and/or nutraceutical formulations of the invention can be administered simultaneously or sequentially with other drugs or biologically active agents. Examples include, but are not limited to antioxidants, free radical scavenging agents, analgesics, anesthetics, anorectals, antihistamines, anti-inflammatory agents including non-steroidal anti-inflammatory drugs, anti-cancer actives, antibiotics, antifungals, antivirals, antimicrobials, antineoplastics, biologically active proteins and peptides, enzymes, hemostatics, steroids including hormones and corticosteroids, etc.

2. Therapeutic Methods And Dosage

[00107] Preferred unit dosage formulations are those containing a daily dose or unit, daily subdose, or an appropriate fraction thereof, of an agent. Therapeutic amounts can be empirically determined and will vary with the pathology being treated, the subject being treated, and the efficacy and toxicity of the agent. Similarly, suitable dosage formulations and methods of administering the agents can be readily determined by those of ordinary skill in the art.

[00108] In some embodiments, a therapeutic method of the present invention can include treating a disease, condition, or disorder by administering to a subject having such disease or condition a stable formulation as described herein in an amount effective to treat the disease, condition, or disorder. In some embodiments, the subject is administered a stable formulation comprising the compounds claimed herein. The disease, condition, or disorder can be a Staphylococcal infection. For prophylactic administration, the composition can be administered to a patient at risk of developing one of the previously described conditions.

[00109] The amount of composition administered will depend upon a variety of factors, including, for example, the particular indication being treated, the mode of administration, whether the desired benefit is prophylactic or therapeutic, the severity of the indication being treated and the age and weight of the patient, etc. Determination of an effective dosage is well within the capabilities of those skilled in the art. In some aspects of the invention, total dosage amounts of a compound composition will typically be in the range of from about 0.0001 or 0.001 or 0.01 mg/kg of patient/day to about 100 mg/kg patient/day, but may be higher or lower, depending upon, among other factors, the activity of the components, its bioavailability, the mode of administration and various factors discussed above. Dosage amount and interval can be adjusted individually to provide plasma levels of the compound(s) which are sufficient to maintain therapeutic or prophylactic effect. For example, the compounds can be administered once per week, several times per week (e.g., every other day), once per day or multiple times per day, depending upon, among other things, the mode of administration, the specific indication being treated and the judgment of the prescribing physician. Skilled artisans will be able to optimize effective local dosages without undue experimentation.

3. Secondary and Combination Treatments

[00110] Certain embodiments provide for the administration or application of one or more secondary or additional forms of therapies or preventative interventions. The type of therapy is dependent upon the type of disease that is being treated or prevented. The secondary form of therapy may be administration of one or more secondary pharmacological agents that can be applied in the treatment or prevention of a Staphylococcal infection.

[00111] The secondary or additional therapy may be administered prior to, concurrently, or following administration of the compositions disclosed herein. The interval between administration of the compositions and the secondary or additional therapy may be any interval as determined by those of ordinary skill in the art. For example, the compositions and the secondary or additional therapy may be administered simultaneously, or the interval between treatments may be minutes to weeks. In embodiments where the agents are separately administered, one would generally ensure that a significant period of time did not expire between the time of each administration, such that each therapeutic agent would still be able to exert an advantageously combined effect on the subject. For example, the interval between therapeutic agents may be about 12 h to about 24 h of each other or within about 6 hours to about 12 h of each other. In some situations, it may be desirable to extend the time period for treatment significantly, however, where several days (2, 3, 4, 5, 6 or 7) to several weeks (1, 2, 3, 4, 5, 6, 7 or 8) lapse between the respective administrations. In some embodiments, the timing of administration of a secondary therapeutic agent is determined based on the response of the subject to the composition. [00112] Examples of secondary treatments useful with methods disclosed herein are: drugs and/or treatments capable of treating and/or preventing a Staphylococcal infection. Further, secondary treatments can include treatments for the symptoms of a Staphylococcal infection or treatment of a Staphylococcal infection or the complications caused thereby. Secondary treatments can also include dietary supplements such as vitamins C, E and D, calcium, zinc, selenium, curcumin, folate, bioflavonoids, resveratrol, and plant extracts. \

H. Kits

[00113] In another aspect of the present invention, kits for treating or preventing a disease, condition or disorder as described herein. For instance, compositions of the present invention can be included in a kit. A kit can include a container. Containers can include a bottle, a metal tube, a laminate tube, a plastic tube, a dispenser, a straw, a pressurized container, a barrier container, a package, a compartment, or other types of containers such as injection or blow-molded plastic containers into which the dispersions or compositions or desired bottles, dispensers, or packages are retained. The kit and/or container can include indicia on its surface. The indicia, for example, can be a word, a phrase, an abbreviation, a picture, or a symbol.

[00114] The containers can dispense a predetermined amount of the composition. In other embodiments, the container can be squeezed ( e.g ., metal, laminate, or plastic tube) to dispense a desired amount of the composition. The composition can be dispensed as a spray, an aerosol, a liquid, a fluid, a semi-solid, or a solid. In a preferred embodiment, the composition is dispensed as a tablet or lozenge. The containers can have spray, pump, or squeeze mechanisms. A kit can also include instructions for employing the kit components as well the use of any other compositions included in the container. Instructions can include an explanation of how to apply, use, and maintain the compositions. The compositions can, if desired, be presented in a pack or dispenser device, which can contain one or more unit dosage forms containing the compound compositions. The pack can, for example, comprise metal or plastic foil, such as a blister pack. The pack or dispenser device can be accompanied by instructions for administration.

EXAMPLES

[00115] The present invention will be described in greater detail by way of specific examples. The following examples are offered for illustrative purposes, and are not intended to limit the invention in any manner. Those of skill in the art will readily recognize a variety of noncritical parameters which can be changed or modified to yield essentially the same results.

EXAMPLE 1

(EXTRACTION OF COMPOUNDS FROM GREEN TEA)

[00116] The inventors have surprisingly found that one or more of several compounds found in green tea can treat and/or prevent a Staphylococcal infection. The inventors have also found that specific relative concentrations of the compounds enhance the ability of the combined compounds to prevent and treat a Staphylococcal infection. In addition, the inventors expect that using compounds of the present invention with additional antibiotics may enhance the ability of the combined compounds to prevent and treat a Staphylococcal infection or reduce the amount of the antibiotic needed. Specifically, it has been shown that the compositions herein when combined with a b-lactam, such as oxacillin, act synergistically against a Staphylococcal infection, such as a MRSA infection.

[00117] Compounds isolated from green tea: Dried Camellia sinensis plants, preferably leaves, were mixed with purified water and let sit for two hours. The mixture was extracted with short chain alcohol, such as food grade ethanol, multiple times. After each extraction, the alcohol phase was discarded and the aqueous phase was retained. The aqueous phase was concentrated, for example using a filtration device with a low protein binding cellulose membrane, or alternatively, polypropylene, Teflon, or nylon membrane can be used. The purpose of filtration was to remove water from the material. The retentate from the filtration was concentrated to dryness at a temperature below 50 °C. The dried material was further chromatographed on a column, preferably 50pm irregular silica. The fraction material was then concentrated to dryness using a suitable drying method, such as rotary evaporator, at a temperature below 50°C, other suitable drying methodologies include, but are not limited to, vacuum drying and spray drying.

EXAMPLE 2

(CHARACTERIZATION OF COMPOUNDS BY ACCURATE MASS AND RELATIVE ABUNDANCE)

[00118] In some aspects, the compounds of the present invention include biomarker compounds defined by compounds found in green tea with an accurate mass of 188.1020 amu, 126.0306 amu, 152.0233 amu, 180.0641 amu, 290.0790 amu, 306.0739 amu, and/or 360.1328 amu. These compounds may be produced synthetically or isolated from an organism such as, but not limited to, green tea. The compounds may be characterized by methods known by one of skill in the art.

[00119] Accurate mass and relative abundances described herein are based on experiments using particular instruments and particular settings and can change from instrument to instrument. There is variability in each measurement. Thus, the accurate mass and relative abundances are defined as being close to as understood by one of ordinary skill in the art. In some non-limiting embodiments the terms are defined to be within 20%, within 10%, within 5%, within 1%, or within 0.5%. In some non-limiting embodiments, the accurate mass has an error of within +/- 20 mmu, within 10 mmu, within 5 mmu, and within 1 mmu. In some non-limiting embodiments, the relative abundance has an error of +/- 20%, within 10%, within 5%, and within 1%, or within 0.5%.

[00120] Methods for Accurate mass: The compounds were characterized and relative abundance was determined using Direct Analysis in Real Time (DART) ion source combined with Time of Flight/Mass Spectrometry (TOF-MS). Specifically, the DART TOF-MS was a JEOL DART™ AccuTOF-mass spectrometer from Jeol USA of Peabody, MA (JMS- T100LC). The mass of the compounds were determined in green tea extract sample by directly introducing the sample to the ion stream by using the closed end of a borosilicate glass melting point capillary tube until a signal was achieved in the total-ion chromatogram (TIC). The next sample was introduced when the TIC returned to baseline levels..

[00121] The settings for the DART ion source were the following:

Gas: He

Flow: 3.98 L/M @ 50 PSI

Temperature: 300 C

Needle Voltage: 3500V

Grid Electrode Voltage: 150V

Discharge Electrode Voltage: 250V

[00122] The settings for the JEOL AccuTOF MS were the following:

Peak Voltage: 1000V

Orifice 1 Voltage: 20V

Ring Lens Voltage: 5V

Orifice 2 Voltage: 5V.

Detector Voltage: 2550V [00123] Calibrations were performed internally with each sample using a 10% (w/v) solution of PEG 600 (Ultra Chemical, North Kingston, RI) that provided mass markers throughout the required mass range 100-1000 m/z. Calibration tolerances were held to 10 mmu. The spectrometric measurements were executed in positive ion mode (M+H]⁺.

[00124] Extract samples were analyzed in six replicates by DART-TOF MS. These six replicates were analyzed to create a single, averaged, filtered, and statistically significant DART fingerprint of the extract. This processed fingerprint was then used to determine the presence of the bioactive markers by comparison of masses. Due to the initial discovery and identification of these bioactive markers, a simple mass comparison was sufficient to determine their presence in any extract or mixture of chemicals. For the AccuTOF, that mass tolerance is less than 20 millimass units (mmu) (predicted mass +/-10 mmu). Given the same extract and ion source, other TOF mass spectrometers may have a higher or lower mass tolerance.

[00125] Methods for Relative Abundance: While no sample preparation is required for a simple analysis with the DART, a curcumin doped/spiked solution was used for determining relative abundance of test compositions through quantitation relative to a known quantity. Standards that are well known and that exist naturally in green tea would vary given any number of influences - growing conditions, harvest time, plant health, etc. For purposes of quantifying the biomarkers, the natural variations of naturally occurring standards make it unacceptable to use as a basis for an absolute quantification of the biomarkers. In order to remove that inconsistency, a compound that is not native to green tea (in this case, curcumin) was used as the basis for a quantitative chemical profile of the bioactive molecules.

[00126] For determining relative abundance of samples with unknown concentrations of the biomarkers disclosed herein, samples of the disclosed compositions were doped/spiked with curcumin. Samples were then analyzed by the DART-TOF method used above.

[00127] Table 1 discloses the relative abundance of the biomarkers disclosed herein found in some non-limiting embodiments of compositions comprising all seven biomarkers found in green tea. Table 1. Relative Abundance of the Biomarkers in Preferred Active Compositions Determined Using 1 mg/ml of the Green tea Composition Spiked with 0.01 mg/ml Curcumin.

EXAMPLE 3

(FORMULATION FOR EXAMPLE 4)

[00128] A dose-reliable, green tea extract comprising biomarkers 1 through 7 was produced in general according to the methods described in Example 1. In powder form, the extract is orally bioavailable and can be stored at room temperature (68-77°F) (data not shown). Current data indicates a shelf life of at least two years (data not shown).

[00129] Dose reliability was tested using an HPLC analytical method to quantify components in the composition of Example 1. The solvent gradient and time program used is in Table 2. The method was established and chosen based on the optimized separation of the botanical reference standards. HPLC analysis of 6 different batches of the composition of Example 1 demonstrate API dose consistent quality standards (FIG. 1).

Table 2. Time program of HPLC solvent system (A=water, B=ACN).

EXAMPLE 4

(STUDIES FOR TREATMENT AND PREVENTION OF A STAPHYLOCOCCAL INFECTION)

[00130] Inhibition and prevention of a Staphylococcal infection by the compounds of the present invention was indicated by multiple studies disclosed herein. It was found that the compounds of the present invention can treat and/or prevent a Staphylococcal infection. Treatment with the compounds of the present invention show activity against MRSA, show no development of resistance, and act synergistically when combined with a b-lactam antibiotic. Specifically, the composition of Example 1 has bactericidal activity and bacteriostatic activity against Staphylococcus and MRSA, provides anti-bacterial activity without causing resistance to the composition, and provides a synergistic effect with oxacillin.

[00131] Optical Density Screening:

[00132] The composition of Example 1 demonstrated significantly more effectiveness against MRSA than clinical doses of oxacillin in an optical density (OD) screening assay (FIG. 2).

[00133] Growth Inhibition:

[00134] Growth inhibition of MRSA and S. aureus was confirmed by incorporating the composition of Example 1 directly into agar growth medium. This method was selected due to the poor diffusability of the composition of Example 1 in the traditional disk diffusion method relative to the growth inhibition seen in the liquid medium. Agar was treated with 1.0, 0.5, 0.25 mg/mL of the composition of Example 1, and MRSA or S. aureus was added at an OD600 of 0.5 to each treatment well. The wells were incubated at 37°C for l8-24h. As controls, MRSA was tested against agar containing 10 pg/ml chloramphenicol and S. aureus was tested against agar containing 25 pg/ml methicillin. The tested concentrations in the treated agar assays describe the concentration over the entire agar plug. The effective concentration at the surface that comes in direct contact with the bacteria is much lower than these concentrations.

[00135] No growth was observed at any of the composition of Example 1 treatment concentrations against either S. aureus or MRSA (FIG. 3a). The composition of Example 1 was as effective an inhibitor as the antibiotic controls chloramphenicol and methicillin in side-by-side experiments (FIG. 3b).

[00136] Minimum Inhibitory Concentration Determination and Synergy:

[00137] Though the composition of Example 1 is highly effective at inhibiting MRSA growth alone, testing was done to determine if synergy could be demonstrated by combining the composition of Example 1 and low dose b-lactam antibiotics, thereby potentially reviving obsolete b-lactam treatments and decreasing the use of more toxic and/or higher dose antibiotics currently used to treat MRSA.

[00138] The minimum effective concentration against MRSA of oxacillin and the composition of Example 1 was determined by OD600 growth curves to be 25pg/mL for oxacillin (12.5x the clinical dose for S. aureus ) and 250pg/mL for the composition of Example 1 (FIG. 4a and FIG. 4b).

[00139] The lowest concentration at which a combination of oxacillin and the composition of Example 1 showed synergistic inhibitory action was determined by the checkerboard OD600 growth curve method. The effective oxacillin dose could be reduced to clinical levels (2pg/mL) with the addition of 80pg/mL or greater of the composition of Example 1 (FIG. 5). Using the fractional inhibitory concentration (FIC) calculation, FIC = (CFrC/MICFrC) + (COx/MICOx), where synergy is determined to be an FIC below 0.5, the composition of Example 1 and oxacillin at these concentrations showed an FIC of 0.4, indicating a synergistic effect. Accordingly, this data confirms a combination therapy of the composition of Example 1 and a clinically relevant dose of oxacillin is a powerful and effective synergistic inhibitor of MRSA in vitro and ex vivo.

[00140] Generational/Resistance Studies:

[00141] Misuse or overuse of antibiotics can lead to development of antibiotic resistance, such as demonstrated with b-lactam antibiotics against S. aureus. Testing to determine if MRSA may develop resistance to the composition of Example 1 treatment, generational studies were performed over a 14 day period using sub-MIC levels of the composition of Example 1 at 90pg/mL. At the end of each 12 hour growth cycle, the composition of Example 1 treated MRSA was plated and colonies counted to determine if growth levels had exceeded that of the untreated MRSA control. Over the entire 14 day period, or approximately 600 generations, no resistance was observed (FIG. 6), as opposed to S. aureus, which can develop resistance to b-lactam antibiotics in as little as 5 days or approximately 160 generations.

[00142] Testing to determine if MRSA develops resistance to the combined composition of Example 1 + oxacillin treatment was performed, due to the MRSA capacity for resistance to oxacillin. MRSA showed increased susceptibility to the composition of Example 1 + oxacillin treatment during the first 3 days of treatment then returned to baseline growth levels between days 4 and 8 (FIG. 7). Growth was inhibited again around day 10 of treatment before returning to baseline growth around day 12. Interestingly, MRSA gained no resistance to the composition of Example 1 treatment alone (FIG. 6) suggesting that there are competing growth pressures between the composition of Example 1 sensitizing MRSA to oxacillin and MRSA becoming resistant to oxacillin again without gaining resistance to the composition of Example 1 treatment.

[00143] In vitro Human Serum Studies:

[00144] It has been shown that when a drug or antibiotic is contained in the blood circulatory system, properties of human serum suppress the action and efficacy of the drug, thus effecting patient dosing. To determine if there are suppressive effects of human serum in the composition of Example 1 against MRSA, human serum was added to physiological levels of 52-55% in growth medium and bacterial colonies were counted at the end of a 12 hour growth cycle. MIC levels of oxacillin in human serum were determined to be l50pg/mL, or 6x what was determined in growth medium alone, giving the baseline suppressive effects of human serum on antibiotic treatment (FIG. 8). A similar suppressive effect was seen for the composition of Example 1, suggesting that that the in vitro anti- MRSA combination product treatments (oxacillin and the composition of Example 1) will correlate equivalently in vivo.

[00145] Conclusions:

[00146] Extensive experimentation had demonstrated that the composition of Example 1 inhibits MRSA growth in an effective manner, with no known resistance development over an extended growth period. Further, enhanced MRSA inhibition synergy is created when a clinical dose of the composition of Example 1 is combined with a clinical dose of oxacillin. EXAMPLE 5

(SYNERGY)

[00147] Because of the predicted method of action of the compositions disclosed herein containing compounds found in green tea, it is believed that the compositions will act synergistically with other agents that treat and/or prevent a Staphylococcal infection, especially those that act through a separate mechanism. To determine synergism with other compounds/compositions, the compositions disclosed herein can be tested in combination with one or more agents and/or treatments that prevent and/or treat a Staphylococcal infection. The agents and/or treatments that prevent and/or treat a Staphylococcal infection can include one or more drugs and/or therapies. Combination studies can show competitive, additive, or synergistic interactions for preventing a Staphylococcal infection, treating a Staphylococcal infection, cell viability, cellular toxicity, side effects, etc. of the combination in cell culture, animal studies, human studies, etc. Non-limiting examples of studies can include those described above and herein as well as those known to one of skill in the art. In some non-limiting examples, the combination of the compositions disclosed herein and one or more drugs and/or therapies can show synergistic activity in preventing and/or treating a Staphylococcal infection.

[00148] A combination assay can be performed to determine the competitive, additive, or synergistic interactions of a combination. In some instances, the assay can utilize an interaction matrix commonly used to look at drug interactions and synergy. In some instances, the interaction matrix is used in a prevention and/or treatment study of a Staphylococcal infection induction in an animal model. Briefly, the experiment can have multiple sets of 25 test animals, in each set of 24: 4 treated with a first test compound/composition (such as the compositions disclosed herein containing compounds found in green tea) alone, 4 with a second test compound/composition (such a b-lactam, 1 with no chemistries, and the remaining 16 can be combinations of the first and second test compounds/compositions. 1:4 dilutions of the first test compound/composition from a starting concentration (such as 1 mg/ml for the compositions disclosed herein containing compounds found in green tea) and 1:4 dilutions of the second test compound/composition from a starting concentration (such as 0.1 mg/ml for the b-lactam) can be tested. The Staphylococcal infection can developed in the constant presence of the inhibitory compounds. In this way, the experiment simulates a patient developing a Staphylococcal infection while on prophylactic treatment and tests prevention of developing a Staphylococcal infection by the first test compound/composition alone, the second test compound/composition/therapy alone, and the combination of the two at a range of concentrations. The data can be analyzed with the methodology of Berenbaum to determine competitive, additive, or synergistic interactions (Berenbaum 1977).

[00149] All of the compositions and/or methods disclosed and claimed herein can be made and executed without undue experimentation in light of the present disclosure. While the compositions and methods of this invention have been described in terms of preferred embodiments, it will be apparent to those of skill in the art that variations may be applied to the compositions and/or methods and in the steps or in the sequence of steps of the method described herein without departing from the concept, spirit and scope of the invention. More specifically, it will be apparent that certain agents which are both chemically and physiologically related may be substituted for the agents described herein while the same or similar results would be achieved. All such similar substitutes and modifications apparent to those skilled in the art are deemed to be within the spirit, scope and concept of the invention as defined by the appended claims.

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Claims

1. A composition comprising the following biomarkers:

biomarker 1 having an accurate mass of 188.1020 amu and having a relative abundance of at least 3.66%;

biomarker 2 having an accurate mass of 126.0306 amu and having a relative abundance of at least 13.52%;

biomarker 3 having an accurate mass of 152.0233 amu and having a relative abundance of at least 2.72%;

biomarker 4 having an accurate mass of 180.0641 amu and having a relative abundance of at least 213.52%;

biomarker 5 having an accurate mass of 290.0790 amu and having a relative abundance of at least 5.84%;

biomarker 6 having an accurate mass of 306.0739 amu and having a relative abundance of at least 2.00%; and

biomarker 7 having an accurate mass of 360.1328 amu and having a relative abundance of at least 18.16%,

wherein the biomarkers are found in green tea; and

wherein the relative abundance is relative to 0.01 mg/ml curcumin spiked in 1 mg/ml of the composition.

2. The composition of claim 1, wherein the biomarkers contained therein have a relative abundance of at most:

biomarker 1 of 5.50%;

biomarker 2 of 20.28%;

biomarker 3 of 4.08%;

biomarker 4 of 320.28%;

biomarker 5 of 8.76%;

biomarker 6 of 3.00%; and biomarker 7 of 27.24%,

wherein the relative abundance is relative to 0.01 mg/ml curcumin spiked in 1 mg/ml of the composition.

3. The composition of any of claims 1 to 2, wherein the mass of each biomarker is the mass as determined by a Direct Analysis in Real Time-TOF (DART-TOF) mass spectrometer.

4. The composition of any one of claims 1 to 3, wherein at least one of the biomarker(s) are synthetically obtained.

5. The composition of any one of claims 1 to 4, wherein at least one of the biomarker(s) are isolated from a plant.

6. The composition of claim 5, wherein at least one of the biomarkers(s) are isolated from green tea.

7. The composition of any one of claims 1 to 6, wherein the composition has an at least 90%, preferably at least 95%, or at least 98% batch-to-batch chemical consistency of relative abundance for the biomarkers.

8. The composition of any one of claims 1 to 7, wherein the composition further comprises a preservative.

9. The composition of any one of claims 1 to 8, wherein the composition further comprises an antibiotic.

10. The composition of claim 9, wherein the antibiotic is a b-lactam.

11. The composition of claim 10, wherein the b-lactam is oxacillin.

12. The composition of any one of claims 1 to 11, wherein the composition is formulated for oral administration.

13. The composition of claim 12, wherein the composition is in a lozenge, a powder, a tablet, a gel-cap, a gelatin, a liquid solution, a syrup, an oil, and/or a dissolvable film.

14. The composition of any one of claims 1 to 11, wherein the composition is formulated for topical administration, administration through injection, and/or intranasal administration.

15. The composition of claim 14, wherein the composition is in a liquid solution, emulsion, powder, or patch.

16. The composition of any of claims 1 to 15, wherein the composition is formulated to treat and/or prevent a staphylococcal infection.

17. The composition of any of claims 1 to 16, wherein the composition is formulated to treat and/or prevent a methicillin resistant Staphylococcus aureus (MRSA).

18. The composition of any of claims 1 to 17, wherein the composition comprises at most 200 mg of a b-lactam.

19. A kit comprising:

a first composition comprising a green tea extract formulated to be administered to a subject; and

a second composition comprising at least one antibiotic formulated to be administered to a subject.

20. The kit of claim 19, wherein the green tea extract is an aqueous extract.

21. The kit of claim 19, wherein the first composition comprises any one of the compositions of claims 1 to 18.

22. The kit of any of claims 19 to 21, wherein the at least one antibiotic is a b-lactam.

23. The kit of any of claims 19 to 22, wherein the kit further comprises an additional antibiotic.

24. A method of treating a subject at risk for or having a staphylococcal infection, the method comprising administering a composition containing a green tea extract to the subject, wherein at least one symptom of the a staphylococcal infection is ameliorated in the subject and/or the onset of the staphylococcal infection is delayed in comparison to the expected onset of the staphylococcal infection if the patient had not been treated.

25. The method of claim 24, wherein the green tea extract is an aqueous extract.

26. The method of claim 24, wherein the composition containing a green tea extract comprises the composition of any one of claims 1 to 18.

27. The method of any of claims 24 to 26, wherein the subject is treated for a methicillin resistant Staphylococcus aureus (MRSA) infection and/or wherein a MRSA infection is prevented.

28. The method of any of claims 24 to 27, wherein the subject is diagnosed as having a methicillin resistant Staphylococcus aureus (MRSA) infection.

29. The method of any one of claims 24 to 28, wherein the subject is administered a total amount of between 1 and 10,000 mg, between 10 and 5,000 mg, between 50 and 2,500 mg, or between 100 and 1,000 mg of the green tea extract during a 24 hour period.

30. The method of any one of claims 24 to 29, wherein the composition containing the green tea extract is administered orally.

31. The method of any one of claims 24 to 29, wherein the composition containing the green tea extract is administered topically.

32. The method of any of claims 24 to 31, wherein the method further comprises administering at least one antibiotic to the subject.

33. The method of claim 32, wherein the at least one antibiotic is a b-lactam.

34. The method of any one of claims 32 to 33, wherein a second composition comprises the at least one antibiotic.

35. The method of any of claims 33 to 34, wherein the subject is administered 200 mg or less of the b-lactam in a 24 hour period.

36. A method of producing a composition of any of claims 1 to 18, wherein the method of producing produces a composition having an at least 90%, preferably at least 95% or at least 98% batch-to-batch chemical consistency of relative abundance for the biomarkers.