METHODS FOR IMPROVING LUNG CONDITION AND AIRWAY

Abstract

The present invention provides methods for improving the lung condition of a cystic fibrosis patient, or for treating a clinical symptom in a patient's airway (e.g., nose, lung, and sinus), by administering a composition comprising an iodine-containing compound and a steroid to the patient.

Claims

1. A method for treating a clinical symptom of the airway of a patient, comprising administering to a sinal cavity of the patient in need therefore a pharmaceutical composition comprising an iodine-containing compound and a steroid, wherein the steroid comprises fluticasone or budesonide, or a salt or ester thereof.

2. The method of claim 1, wherein the sinus symptom is inflammation, infection, formation of biofilm, rhinosinusitis, congestion, pain, pressure, fatigue, or thickened nasal discharge.

3. The method of claim 1, wherein the airway is sinus, nose, or lung.

4. The method of claim 1, wherein the pharmaceutical composition is formulated as an aqueous solution or a suspension; and the pH of the aqueous solution or the suspension ranges from about 2.0 to about 7.5, from about 2.0 to about 7.0, from about 3.0 to about 6.0, or from about 4.0 to about 5.0.

5. The method of claim 1, wherein the iodine-containing compound comprises povidone-iodine.

6. The method of claim 1, wherein the dose volume of the iodine-containing compound administered to the subject is between about 10 mcg to about 300 mcg per day, about 20 mcg to about 200 mcg per day, about 30 mcg to about 100 mcg per day, or about 50 mcg per spray with two spray per nostril per day.

7. A method for improving the lung condition of a cystic fibrosis patient, comprising washing the sinal cavity of the patient with a pharmaceutical composition comprising an iodine-containing compound and a steroid, wherein the steroid comprises mometasone, fluticasone or budesonide, or a salt or ester thereof.

8. The method of claim 7, wherein the iodine-containing compound comprises povidone-iodine.

9. The method of claim 7, wherein the concentration range of the iodine-containing compound is from 0.01% to 0.2% by weight/weight or weight/volume percentage, between 0.2% and 1.0% by weight/weight or weight/volume percentage, 0.15% by weight/weight or weight/volume percentage, or 15% by weight/weight or weight/volume percentage.

10. A method for stabilizing an iodine-containing compound at a low concentration, comprising mixing the iodine-containing compound with a steroid, wherein the steroid comprises fluticasone or budesonide, or a salt or ester thereof.

11. The method of claim 10, wherein the iodine-containing compound comprises povidone-iodine.

12. The method of claim 11, wherein the concentration of the povidone-iodine ranges from about 0.01% to about 10%, from about 0.1% to about 2.5%, from about 0.15% to about 1.5%, from about 0.2% to about 1.0%, or about 0.2% by weight/weight percentage or weight/volume percentage.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0041] In one aspect, the invention provides surprisingly stable pharmaceutical compositions for treating a sinal symptom which comprises an iodine-containing compound (e.g., povidone-iodine) and a steroid (budesonide, micronized fluticasone proprionate, or micronized mometasone).

[0042] In an embodiment, the invention provides a preserved steroid composition comprising a povidone-iodine composition. In an embodiment, the concentration of povidone-iodine as a preservative in steroid compositions can range from 0.01-10% (weight/weight or weight/volume), and all concentrations in between. In an embodiment, the povidone-iodine concentration is between 0.1% and 2.5%, in another embodiment, between 0.15 and 1.5%, and in yet another embodiment, between 0.2% and 1.0%. In an embodiment, the povidone-iodine concentration is about 0.2%.

[0043] These pharmaceutical compositions (e.g., containing povidone iodine and a steroid) possess antimicrobial property. Thus, in another aspect, an iodine-containing compound (e.g., povidone iodine) provides a steroid preparation with one or more non-antimicrobial preservative properties (e.g., antioxidant). In an embodiment, the invention also provides povidone-iodine compositions comprising one or more components in addition to the povidone-iodine component, as set forth herein. In an aspect, the invention provides a broad spectrum of povidone-iodine steroid compositions. In an embodiment, a preparation comprises an anti-inflammatory compound selected from the group consisting of budesonide, fluticasone, mometasone, all salts thereof, all esters thereof, and any combination thereof.

[0044] It is well known to those skilled in the art that more dilute solutions of PVP-I are more effective anti-microbial agents. It is useful to supply these PVP-I solutions to areas of infection and/or inflammation in the concentration range around 0.01% PVP-I (w/w) and about 1% PVP-I. However, the aqueous stability of PVP-I in dilute concentrations below 0.40% limits their long term storage. The iodine component of these dilute PVP-I preparations can often decay when the starting PVP-I concentration is much less than 0.4% (see, e.g., U.S. Pat. Nos. 4,976,969 and 4,996,048). We have developed a stable, chemically distinct suspension of PVP-I and steroid in small aqueous volumes for dilution into irrigating solutions to be employed in sinonasal irrigation. In this way we have developed stable solutions of PVP-I/steroids that can be used to deliver therapeutic steroid concentrations along with low-concentration PVP-I for therapeutic treatment of acute and chronic sinus infections. The compositions do not suffer from the long term instability of PVP-I low concentration preparations and yet still deliver an effective lower concentration of PVP-I to the intended area.

[0045] One example of the pharmaceutical compositions described above includes 400 mg PVP-I USP grade dry powder dissolved in 4 cc sterile water into which is suspended 0.6 mg of budesonide. This is supplied as a stable 4 cc vial for patients to keep in their homes. Prior to irrigation into the sinuses, approximately 200 cc of water is added to an 8 oz sinus irrigation bottle to make an irrigating solution of approximately 0.2% PVP-I aqueous concentration and still contains 0.6 mg suspended budesonide. In other examples, the steroid can be mometasone, fluticasone or other nasal steroids and the PVP-I can vary between 0.005 g and 1 g dissolved in 1 cc, 2 cc, 3 cc, 4 cc or any other small volume of suspendant suitable to suspend the steroid.

[0046] Additionally, non-limiting examples of suitable excipients for the pharmaceutical compositions of this invention include co-solvents, surfactants, viscosity agents, or bioadhesive agents. Furthermore, the compositions can be combined with an effective amount of a chemical agent to provide a cooling sensation to relieve mild irritation, enhance comfort, provide a refreshing effect, and improved sensation, when the povidone-iodine solution is applied. Such an agent encompasses various chemicals and chemical classes, including, but not limited to, cooling agents such as menthol, menthol derivatives including methone glycerin acetyl and menthyl esters, carboxamides, menthane glycerol ketals, alkyl substituted ureas, sulfonamides, terpene analogs, furanones, and phosphine oxides; or camphor, and borneol.

[0047] In an aspect, the compositions of the invention may optionally comprise a co-solvent. In an embodiment, the solubility of the components of the compositions may be enhanced by a surfactant or other appropriate co-solvent in the composition. Such co-solvents/surfactants include, but are not limited to, polysorbate-20, -60, and -80, polyoxyethylene/polyoxypropylene surfactants (e.g. Pluronic F-68, F-84 and P-103), cyclodextrin, tyloxapol, PEG 35 Caster oil (Cremophor EL), polyoxyl 40 Stearate (Myrj 52), as well as other agents known to those skilled in the art, or any combination thereof. Typically, such co-solvents are employed at a level of from about 0.01% to about 2% by weight.

[0048] In another aspect, the compositions of the invention may optionally comprise an optional viscosity-increasing agent. Viscosity increased above that of simple aqueous solutions may be desirable to increase absorption of the active compound, to decrease variability in dispensing the formulation, to decrease physical separation of components of a suspension or emulsion of the formulation and/or to otherwise improve the irrigating steroid formulation. Such viscosity-enhancing agents include, but are not limited to, polyvinyl alcohol, polyvinylpyrrolidone, methyl cellulose, hydroxypropylmethylcellulose, hydroxyethylcellulose, carboxymethylcellulose, hydroxypropylcellulose, microcrystalline cellulose, carboxymethyl cellulose sodium, dextrose anhydrous, other agents known to those skilled in the art, and/or any combination thereof. Such agents are typically employed at a level of from 0.01% to 2% by weight.

[0049] In another aspect, bioadhesive agents may comprise the compositions, in order to increase the retention time of the drug gradient over a biological substrate. The bioadhesive agents include, but are not limited to: polyvinylpyrrolidone (PVP), xanthan gum, locust bean gum, acacia gum, hydroxypropyl methylcellulose (HPMC), sodium alginate, pectin, gelatin, carbomer, polyvinylalcohol, gellan gum, tragacanth, acacia, and sodium carboxymethyl cellulose, as well as other agents known to those skilled in the art, or any combination thereof.

[0050] The povidone-iodine-comprising steroid composition may be in the form of a solution, a suspension, an emulsion, a dry sterile powder, or a controlled-release/sustain-release vehicle. It may be applied to a subject through metered-dose, manual-pump spray, or a metering, atomizing spray pump, or any inhalation device.

[0051] In any of the compositions of this disclosure for sinonasal inhalation administration, the mixtures are preferably formulated as aqueous solutions or suspensions at a pH in the range of 2.0 to 7.0. It will be understood that a range listed herein is intended to encompass the upper and lower bounds of the range, inclusively. In an embodiment, the pH is in the range of 4 to 5. This pH range may be achieved by the addition of acids/bases to the solution. In another embodiment, the pH is in the range of 3 to 6. In another embodiment, the pH is in the range of 3-6 and can be maintained at selected pH ranges of 1-2 pH unit from 2.0 to 7.5 through the use of appropriate buffers.

[0052] The invention also provides methods of using the pharmaceutical compositions described herein for treating a clinical symptom of the airway of a patient, e.g., by administering the compositions in the patient's sinal cavity. In an embodiment, the dose volume administered to the patient may range from about 10 mcg to about 300 mcg per day, from about 20 mcg to about 200 mcg per day, from about 30 mcg to about 100 mcg per day, or about 50 mcg per spray, two sprays per nostril once daily.

[0053] As used herein, the term “or” is meant to include both “or” and “and.”

[0054] As used herein, the term “pharmaceutical compositions” is interchangeable with pharmaceutical preparation.

[0055] As used herein, the term “a patient” refers to a mammal, including animal or human being.

[0056] As used herein, the term “iodine-containing compound” is meant to include free iodine, complex or adduct containing iodine and capable of releasing iodine under certain conditions.

[0057] As used herein, the term “steroid” refers to anti-inflammatory steroids including corticosteroids. Specific examples include hydrocortisone, hydrocortisone acetate, cortisone acetate, tixocortol pivalate, prednisolone, methylprednisolone, and prednisone; triamcinolone acetonide, triamcinolone alcohol, mometasone, amcinonide, budesonide, desonide, fluocinonide, fluocinolone acetonide, and halcinonide; betamethasone, betamethasone sodium phosphate, dexamethasone, dexamethasone sodium phosphate, and fluocortolone; hydrocortisone-17-butyrate, hydrocortisone-17-valerate, aclometasone dipropionate, betamethasone valerate, betamethasone dipropionate, prednicarbate, clobetasone-17-butyrate, clobetasol-17-propionate, fluocortolone caproate, fluocortolone pivalate, and fluprednidene acetate; flunisolide; fluticasone propionate; triamcinolone acetonide; beclomethasone dipropionate, and budesonide.

[0058] As used herein, the term “tonicity agent” refers to an agent used to adjust the composition of the formulation to the desired isotonic range. Examples of tonicity agents include glycerin, mannitol, sorbitol, sodium chloride, and other electrolytes. See, e.g., U.S. Pat. No. 6,933,289.

[0059] As used herein, the term “surfactant” refers to a compound that lowers the surface tension of a liquid, the interfacial tension between two liquids, or that between a liquid and a solid. Some commonly encountered surfactants include: alkyl sulfates (e.g., ammonium lauryl sulfate, sodium lauryl sulfate); alkyl ether sulfates (e.g., sodium laureth sulfate, also known as sodium lauryl ether sulfate (SLES), sodium myreth sulfate); sulfonates; docusates (e.g., dioctyl sodium sulfosuccinate); sulfonate fluorosurfactants (e.g., perfluorooctanesulfonate, perfluorobutanesulfonate); alkyl benzene sulfonates; alkyl aryl ether phosphate; alkyl ether phosphate; alkyl carboxylates (e.g., fatty acid salts (soaps): sodium stearate); sodium lauroyl sarcosinate; carboxylate fluorosurfactants (e.g., perfluorononanoate, perfluorooctanoate); octenidine dihydrochloride; alkyltrimethylammonium salts (e.g., cetyl trimethylammonium bromide, cetyl trimethylammonium chloride); cetylpyridinium chloride; polyethoxylated tallow amine; benzalkonium chloride; benzethonium chloride; 5-bromo-5-nitro-1,3-dioxane; dimethyldioctadecylammonium chloride; dioctadecyldimethylammonium bromide; (3-[(3-Cholamidopropyl) dimethylammonio]-1-propanesulfonate); sultaines (e.g., cocamidopropyl hydroxysultaine); amino acids; imino acids; betaines (e.g., cocamidopropyl betaine); fatty alcohols (e.g., cetyl alcohol, stearyl alcohol, cetostearyl alcohol; oleyl alcohol); polyoxyethylene glycol alkyl ethers; octaethylene glycol monododecyl ether, pentaethylene glycol monododecyl ether; polyoxypropylene glycol alkyl ethers; glucoside alkyl ethers; decyl glucoside, lauryl glucoside, octyl glucoside; polyoxyethylene glycol octylphenol ethers; Triton X-100; polyoxyethylene glycol alkylphenol ethers; nonoxynol-9; glycerol alkyl esters; sorbitan alkyl esters; cocamide MEA, cocamide DEA; dodecyldimethylamine oxide; block copolymers of polyethylene glycol and polypropylene glycol.

[0060] As used herein, the term “treating” refers to reducing, either partially or completely, the severity of a symptom shown by a patient.

[0061] All references cited are incorporated herein by reference in their entireties.

[0062] The invention is now described with reference to the following Examples. These Examples are provided for the purpose of illustration only and the invention should in no way be construed as being limited to these Examples, but rather should be construed to encompass any and all variations which become evident as a result of the teaching provided herein.

EXAMPLE 1

Preparation of PVP-I (0.2%), Budesonide (0.6 mg) Nasal Irrigation

[0063] 400 mg PVP-I USP grade dry powder was dissolved in 4g sterile water into which is suspended 0.6 mg of budesonide suspension, which may have or have not contained other excipients such as co-solvents/surfactants, bioadhesives, and tonicity agents. Prior to irrigation into the sinuses, approximately 200 mL of water was added to an 8 oz sinus irrigation bottle to QS 240 mL and made an irrigating solution of 0.2% PVP-I aqueous concentration and still contained 0.6 mg suspended budesonide.

EXAMPLE 2

Preparation of PVP-I, Budesonide Suspension, 1.0 mg

[0064] Budesonide suspension with PVP-I concentration ranging from about 0.25% by weight was prepared as set forth herein. By way of a non-limiting example, a composition was prepared using approximately 0.6 g PVP-I product, and combining with micronized budesonide 1.0 mg, microcrystalline cellulose and carboxymethyl cellulose sodium, polysorbate 80, disodium edetate, sodium chloride, and purified water; hydrochloric acid and/or sodium hydroxide was added to adjust the pH to a target of 4-6. To the above mixture was added water QS 240 mL. The resulting mixture was a 0.25% PVP-I, isotonic irrigating mixture that delivers a total daily dose of 1.0 mg budesoinde via sinonasal irrigation when completely administered in one day. The preparation was suitable for sinonasal irrigation or topical administration.

EXAMPLE 3

Preparation of PVP-I, Fluticasone Proprionate Suspension, 1.0 mg

[0065] Budesonide suspensions with PVP-I concentration ranging from about 0.25% by weight were prepared as set forth herein. By way of a non-limiting example, a composition was prepared using approximately 0.6 g PVP-I product, and combining with micronized fluticasone proprionate 1.0 mg, microcrystalline cellulose and carboxymethyl cellulose sodium, polysorbate 80, disodium edetate, sodium chloride, and purified water; hydrochloric acid and/or sodium hydroxide is added to adjust the pH to a target of 4-6. To the above mixture was added water QS 240 mL. The resulting mixture was a 0.25% PVP-I, isotonic irrigating mixture that delivered a total daily dose of 1.0 mg fluticasone proprionate via sinonasal irrigation when completely administered in one day. The preparation was suitable for sinonasal irrigation or topical administration.

EXAMPLE 4

Preparation of PVP-I, Mometasone Suspension, 1.0 mg

[0066] Budesonide suspension with PVP-I concentration ranging from about 0.25% by weight is prepared as set forth herein. By way of a non-limiting example, a composition is prepared using approximately 0.6 g PVP-I product, and combining with micronized mometasone 1.0 mg, microcrystalline cellulose and carboxymethyl cellulose sodium, polysorbate 80, disodium edetate, sodium chloride, and purified water; hydrochloric acid and/or sodium hydroxide is added to adjust the pH to a target of 4-6. To the above mixture is added water QS 240 mL. The resulting mixture is a 0.25% PVP-I, isotonic irrigating mixture that delivers a total daily dose of 1.0 mg mometasone via sinonasal irrigation when completely administered in one day. The preparation is suitable for sinonasal irrigation or topical administration.

EXAMPLE 5

Antimicrobial Activity of PVP-I Preserved Steroid Solutions

[0067] By way of a non-limiting example, PVP-I Preserved Steroid Solutions were prepared according to methods described in Examples 1-4. These solutions were then tested for in-vitro microbiological activity. Microbiological activity can be tested for antimicrobial activity against, for example, bacteria found in the mouth (P. gingivalis), or against other bacteria. In another example, killing time tests were conducted with a series of log phase cultures of gram negative and gram positive organisms including Gentamicin resistant Pseudomonas aerouginosa, methicilin-resistant staph aureus, E. coli, chlamydia trachoma and selected viruses including adenoviruses and rhinoviruses. Controls used may include steroid preparations of commercially available antimicrobial products. Bacterial samples were taken at 30 seconds, 1, 2, 5, 10 and 15 minutes and transferred into culture media containing inactivators for iodine. Similarly, virus killing time tests were sampled at one minute and transferred into inactivating media. The results obtained with the experimental samples were compared with the control samples to assess the level of antimicrobial activity of a composition of the invention.

EXAMPLE 6

Antimicrobial Preservative Effectiveness Test of PVP-I In Nasal Steroid Preparations

[0068] By way of a non-limiting example, PVP-I Preserved Steroid Solutions were prepared according to methods described in Examples 1-4. These solutions were then tested for preservative effectiveness according to standard procedures described in the United States Pharmacopeia, General Chapter 51. The iodophor preservatives as prepared in Examples 1-4 were employed in such a way as to satisfy all requirements for preservative effectiveness as described by USP <51>.

EXAMPLE 7

Preparation of PVP-I (10%), Budesonide (0.3 g) Nasal Irrigation

[0069] 200 g PVP-I USP grade dry powder was dissolved in 4 g sterile water into which was suspended 0.3 g of budesonide suspension, then added 0.32 g polysorbate 80, 18 g NaCl, respectively. Water was added to the above mixture to 2000 g to give rise to an irrigating solution of 10% PVP-I aqueous concentration, with 0.3 g suspended budesonide.

[0070] Stability of the Solutions: The amount of titratable iodine in the solutions was determined by titration method after 6 months of sample storage at different temperatures.

[0071] Titration Method: 5 mL of each sample was transferred into a 125 mL beaker by pipette, and 1 mL of 1% (w/v) starch indicator solution was added. The solution was titrated with 0.02 N sodium thiosulfate solution until the blue color disappeared completely. The volume of sodium thiosulfate solution used was determined.

[0072] Titration Iodine (mg)=0.02N*V (mL, volume used for titration)*12.69(mg/mL)/2

TABLE-US-00001 TABLE 1 Stability Data Summary (Available Iodine at 5° C.) Glass Glass vial vial with with spray spray pump pump cap Glass cap Hori- vial Vertical zontal with Storage Avail- Storage Avail- cap Avail- average able average able average able % w/w iodine % w/w iodine % w/w iodine Day iodine (%) iodine (%) iodine (%) 0 day 0.9719 100.0 0.9719 100.0 0.9719 100.0 15th day 0.9687 99.67 0.9686 99.66 0.9639 99.17 1 month 0.9740 100.22 0.9670 99.50 0.9635 99.14 3 months — — — — 0.9541 98.17 6 months 0.9546 98.22

TABLE-US-00002 TABLE 2 Stability Data Summary (Available Iodine at 25° C.) Glass Glass Vial Vial with with Spray Spray pump pump cap Glass cap Hori- Vial Vertical zontal with Storage Avail- Storage Avail- Cap Avail- average able average able average able % w/w iodine % w/w iodine % w/w iodine Day iodine (%) iodine (%) iodine (%) 0 day 0.9719 100.0 0.9719 100.0 0.9719 100.0 15th day 0.9561 98.37 0.9560 98.36 0.9581 98.58 1 month 0.9782 100.60 0.9751 100.33 0.9768 100.50 2 months — — — — 0.9523 97.98 3 months — — — — 0.9515 97.90 6 months 0.9524 98.01

TABLE-US-00003 TABLE 3 Stability Data Summary (Available Iodine at 30° C.) Glass Glass Vial Vial with with Spray Spray pump pump cap Glass cap Hori- Vial Vertical zontal with Storage Avail- Storage Avail- Cap Avail- average able average able average able % w/w iodine % w/w iodine % w/w iodine Day iodine (%) iodine (%) iodine (%) 0 day 0.9719 100.0 0.9719 100.0 0.9719 100.0 15th day 0.9553 98.29 0.9565 98.42 0.9513 97.88 1 month 0.9592 98.69 0.9600 98.78 0.9577 98.54 3 months — — — — 0.9452 97.05 6 months — —

TABLE-US-00004 TABLE 4 Stability Data Summary (Available Iodine at 40° C.) Glass Glass Vial Vial with with Spray Spray pump pump cap Glass cap Hori- Vial Vertical zontal with Storage Avail- Storage Avail- Cap Avail- average able average able average able % w/w iodine % w/w iodine % w/w iodine Day iodine (%) iodine (%) iodine (%) 0 day 0.9719 100.0 0.9719 100.0 0.9719 100.0 15th day 0.9449 97.22 0.9442 97.15 0.9435 97.06 1 month 0.9544 98.02 0.9567 98.44 0.9531 98.07 2 months — — — — 0.9231 94.98 3 months — — — — 0.9170 94.35

[0073] The data of available iodine content of the formulations after months of storage at different temperatures (5° C., 25° C., 30° C., 40° C.), have suggested that stable combination formulations have been achieved for PVP-Iodine combinations with budesonide. The available iodine content of the formulations at different temperatures showed no significant degradation after 6 months. The data showed excellent stability of the PVP-Iodine.

Stability Data of Budesonide Content in the Sample Using HPLC

[0074] The USP method was performed. The concentration of budesonide data is tabulated as follows.

TABLE-US-00005 TABLE 5 The stability of budesonide at 5° C. (content unit μg/mL): Glass Vial Glass Vial with Spray Budes- with Spray Budes- Glass Budes- pump cap onide pump cap onide Vial onide Vertical content Horizontal content with content Day Storage (%) Storage (%) Cap (%) 0 day 150.0 100 150.0 100 144.0 100 15th day 146.4 97.6 150.7 100.5 146.6 101.8 1 month 146.5 97.7 152.3 101.5 144.8 100.6 3 months — — — — 146.4 101.7 6 months 146.0 101.5

TABLE-US-00006 TABLE 6 The stability of budesonide at 25° C. (content unit μg/mL): Glass Vial Glass Vial with Spray Budes- with Spray Budes- Glass Budes- pump cap onide pump cap onide Vial onide Vertical content Horizontal content with content Day Storage (%) Storage (%) Cap (%) 0 day 150.0 100 150.0 100 144.0 100 15th day 149.1 99.4 148.4 98.9 143.5 99.7 1 month 149.3 99.5 145.7 97.1 145.7 101.2 2 months — — — — 139.0 96.5 3 months — — — — 139.6 97.0 6 months 135.3 94.0

TABLE-US-00007 TABLE 7 The stability of budesonide at 30° C. (content unit μg/mL): Glass Vial Glass Vial with Spray Budes- with Spray Budes- Glass Budes- pump cap onide pump cap onide Vial onide Vertical content Horizontal content with content Day Storage (%) Storage (%) Cap (%) 0 day 150.0 100 150.0 100 144.0 100 15th day 151.6 101.1 149.4 99.6 145.3 100.9 1 month 150.2 100.1 148.2 98.8 142.2 98.8 3 months — — — — 134.1 93.1

TABLE-US-00008 TABLE 8 The stability of budesonide at 40° C. (content units μg/mL): Glass Vial Glass Vial with Spray Budes- with Spray Budes- Glass Budes- pump cap onide pump cap onide Vial onide Vertical content Horizontal content with content Day Storage (%) Storage (%) Cap (%) 0 day 150.0 100 150.0 100 144.0 100 15th day 143.9 95.9 137.8 91.9 139.8 97.0 1 month 139.0 92.7 133.6 89.1 132.3 91.9 2 months — — — — 124.9 86.7 3 months — — — — 115.5 80.2

[0075] The data of concentration of budesonide after months of storage at different temperatures (5° C., 25° C., 30° C., 40° C.), have suggested that stable combination formulations have been achieved for budesonide combinations with PVP-Iodine. The concentration of budesonide at different temperatures showed no significant degradation after 6 months at room temperature. The data showed good stability of the budesonide.

Accelerated Degradation Test of PVP-I in the Sample and in the Control (without Budesonide) at 60° C. Using Titration

[0076]

TABLE-US-00009 TABLE 9 Sample Control average % w/w Available average % w/w Available Day Available iodine iodine (%) Available iodine iodine (%) 0 day.sup.  0.9822 100 1.0064 100 2 days 0.9628 98.02 0.9608 95.46 3 days 0.9385 95.55 0.9284 92.25 8 days 0.9159 93.25 0.9150 90.92

[0077] The data of the accelerated degradation test of available iodine in the PVP-Iodine combination formulation in the sample and in the control sample (without budesonide) at 60° C., have suggested that the sample showed a better stability than the control sample, indicating stable combination formulations have been achieved for budesonide combinations with PVP-Iodine. Therefore, satisfactory stability of available iodine was obtained.

Accelerated Degradation Test of Budesonide Content (HPLC) in the Sample and in the Control (without PVP-Iodine) at 60° C. Using Titration

[0078]

TABLE-US-00010 TABLE 10 Sample Budesonide Control Budesonide Day (μg/mL) content (%) (μg/mL) content (%) 0 day.sup.  140.9 100 143.8 100 2 days 131.2 93.1 133.4 92.8 3 days 118.5 84.1 125.8 87.5 8 days 106.4 75.5 126.3 87.8

[0079] The data of the accelerated degradation test of budesonide in the sample and in the control (without PVP-Iodine)at 60° C., have suggested that the sample showed a comparable stability with the control sample, but during the test, it is surprising that we observed that the sample was much more homogeneous than the control. This illustrated the solubility of the budesonide was significantly improved by addition of PVP-Iodine.

EXAMPLE 8

Studies of Efficacy on Established Bacterial Biofilms

[0080] Preparation of PVP-1/Budesonide Suspension. The test formulation consisted of a 10.0% povidone-iodine/0.6 mg budesonide (PVP-I/B) suspension in 4 mL water (ASL Pharmacy, Camarillo, Calif.). This concentrated 10% PVP-I/B suspension maintained room-temperature stability for at least 3 months and was self-preserved. The 4 mL suspension was diluted prior to testing with an additional 200 mL of water in a procedure identical to the one employed by patients preparing the same dilute suspension for use as sinonasal irrigation. The resulting extremely dilute (0.2%) PVP-I concentration was chosen based on reported antimicrobial efficacy in vitro, the known pharmaceutical chemistry of PVP-I topical solutions and prior safety studies demonstrating no effect on ciliary beat frequency and the absence of cytotoxicity. The concentration of budesonide was chosen to be consistent with published irrigation trials demonstrating safety and efficacy at a similar cumulative daily dose.

[0081] Challenge Organism Preparation: Planktonic bacterial samples of Staphylococcus aureus (ATCC#6538) and Psuedomonas aeruginosa (ATCC# 9027) were obtained from ATCC (Manassas, Va.). A sterile swab of S. aureus or P. aeruginosa was aseptically taken from stock cultures maintained at 2-8° C. and aseptically transferred to sterile Trypytic Soy Agar (TSA) slants. The fresh slants were incubated at 30-35° C. for 18-24 hours. Sterile TS saline 10 mL was pipetted into each slant subsequent to incubation and the growth was mechanically dislodged with a sterile cotton-tipped applicator. The suspension was transferred to a sterile 50 mL polypropylene centrifuge tube, washed by centrifugation at 4000×g for 8-10 minutes, supernatant decanted and pellet resuspended in 10 mL saline TS. The resulting suspension was washed a second time, resuspended in 10 mL saline TS and adjusted to an organism concentration of ≈108 cfu/mL.

[0082] Optimization: Three 50 mL conical tubes were filled with approximately 30 mL TSB inncoulated at a level of ≈105 cfu/mL of each challenge organism Innoculation controls were prepared containing no coupons for growth comparison. Innoculated tubes were placed on an orbital shaker set to 120 rpm and incubated at 30-37±2° C. for 24±4 hours. Following incubation, biofilms were recovered and enumerated as below.

[0083] Sample Preparation and Challenge: Three samples per challenge organism and three controls each were prepared, inoculated and incubated as in the optimization step above. Following incubation the coupons were aseptically removed from their respective 50 mL conical tubes, gently rinsed with sterile TS saline to remove any loosely adhered planktonic cells and placed into new individual sterile 50 mL conical tubes. For each challenge organism, three samples were placed into 10 mL of sterile saline and 3 were challenged with 10 mL of the test PVP-I/B suspension. After 10 minutes of contact time, 10 mL of Dey-Engly Neutralizing Broth (DEB) was added to all tubes to prevent further antimicrobial action.

[0084] Neutralization: For each test suspension, the test product was diluted 1:10 using DEB. A control sample of 10 mL sterile TS saline was prepared for comparison. Both tubes were inoculated with sufficient inoculums to result in 10-100 cfu of challenge organisms per plate. Inoculated tubes were incubated at 10 minutes to allow for neutralization at ambient temperature. Appropriate aliquots from each tube were plated in duplicate with TSA. The recovery in the neutralizer broth was at least 50% of the recovery in the control samples. Following neutralization, samples were recovered and enumerated as below.

[0085] Recovery and Enumeration of Organisms: Following incubation, test samples and controls were removed from their respective tubes, gently rinsed and the organisms removed from the test surfaces and controls through sonication. Serial dilution of the recovered organisms was performed and the recovered organisms were then quantified.

[0086] Results: The log reductions in viable organisms post-exposure for PVP-I/B and saline controls are presented in Table 11 and Table 12. The average log reduction in viable P. aeruginosa from a treated biofilm is 3.9. The average log reduction in viable S. aureus from a treated biofilm is 4.2.

TABLE-US-00011 TABLE 11 Reduction in Recoverable Organisms from P. Aeruginosa Biofilms Saline Exposed Controls PVP-I/ Budesonide LOG Dilution Dilution AVG CFU Corrected LOG CFU Corrected LOG vs Dilution Recovered AVG AVG Dilution Recovered AVG AVG Control 10.sup.−5 29 25 2.7 × 10.sup.6 6.4 10.sup.−2 5 2 350 2.5 3.9 10.sup.−5 18 28 2.3 × 10.sup.6 6.4 10.sup.−2 14 21 1750 3.2 3.2 10.sup.−5 14 12 1.3 × 10.sup.6 6.1 10.sup.−2 9 7 800 2.9 3.2

TABLE-US-00012 TABLE 12 Reduction in Recoverable Organisms from S. Aureus Biofilms Saline Exposed Controls PVP-I/ Budesonide LOG Dilution Dilution AVG CFU Corrected LOG CFU Corrected LOG vs Dilution Recovered AVG AVG Dilution Recovered AVG AVG Control 10.sup.−5 23 27 2.5 × 10.sup.6 6.4 10.sup.−2 1 3 200 2.3 4.1 10.sup.−5 22 24 2.3 × 10.sup.6 6.4 10.sup.−2 2 2 200 2.2 4.1

EXAMPLE 9

Studies of Efficacy on Established Fungal Biofilms

[0087] In a procedure similar to that described in Example 8 (above), the invention was evaluated for efficacy against fungal biofilms established with Candida albicans. The results of the assay for efficacy are summarized in Table 13.

TABLE-US-00013 TABLE 13 Reduction in Recoverable Organisms from C. Albicans Biofilms Saline Exposed Controls PVP-I/ Budesonide CFU Dilution CFU Dilution Re- Corrected LOG Re- Corrected LOG Dilution covered AVG AVG Dilution covered AVG AVG 10.sup.−3 31 29  3.0 × 10.sup.4 4.48 10.sup.−2 0 0 0 NA 10.sup.−3 60 65 6.25 × 10.sup.4 4.79 10.sup.−2 0 0 0 NA 10.sup.−3 27 21  2.4 × 10.sup.4 4.38 10.sup.−2 0 0 0 NA

EXAMPLE 10

Povidone-Iodine Irrigation in Human Subjects

[0088] The study treatment consists of sinonasal irrigation with a 0.2% PVP-I/0.6 mg budesonide (PVP-I/B) suspension in purified water, prepared via dilution of a preparation identical to Example 8 above, administered through a commercially available low-pressure, high-volume bottle (Sinus Rinse, NeilMed, Santa Rosa, Calif.). Treatment was administered BID to each nostril for at least 2 weeks. Pre-treatment bacterial and fungal cultures were obtained and compared to cultures obtained at the conclusion of the study.

[0089] Results. A total of five patients were studied. Patient demographics are detailed in Table 13. None of the patients discontinued use due to intolerance. There were no reported adverse reactions to sinonasal irrigation with PVP-I/B. The mean post-treatment improvement in the subjective symptoms as measured by scaled scores on a validated sinonasal outcomes test was 31. Pre-treatment cultures were positive for 5/5 patients with multi-resistant species including MRSA, Enterococcus, Acenitobacter, Pseudomonas, Propionobacterium, S. viridans, Klebsiella and Serratia. Post-treatment cultures were positive 2/5 patients only for S. aureus, Pseudomonas and Enterococcus (Table 16).

TABLE-US-00014 TABLE 15 Patient Demographics Patient Age Gender Diagnosis Pre-Op Treatment tPVP-I/B 001 22 M CF ESS/PO & IV Abx 4 wks 002 49 F CRS rESS/PO Abx/ 3 wks 003 57 M CRS ESS/PO Abx 3 wks 004 54 F CRS ESS/PO Abx 2 wks 005 62 F CLL/L Chemo/XRT 3 wks CRS: chronic rhinosinusitis. CF: cystic fibrosis. CLL: chronic lymphocytic leukemia. L: lymphoma. ESS: endoscopic sinus surgery. rESS: revision endoscopic sinus surgery. tPVP-I/B duration of treatment with PVP-I/B. PO Abx: oral antibiotics. IV Abx: intravenous antibioitics

TABLE-US-00015 TABLE 16 Clinical and Microbiological Data Patient preCx preSnOT postCx postSnOT 001 MRSA Ps 24 Ps 17 002 KO, Sa, SM 61 Neg 15 003 En, AB 57 En 11 004 MRSA, PA, SV 71 neg 34 005 Sa 53 Sa 36 MRSA: methicillin resistant staph aureaus. Ps: pseudomonas auroginosa. KO: Klebsiella oxytoca. Sa: Staph aureaus. SM: Serratia marcescens. En: Enterococus Cloacae. AB: acenitobacter Baumanni. PA: Propionibacterium Acnes. SV: Strep viridans. Neg: negative.

[0090] We developed our treatment program to address the common features of CRS across the whole spectrum of the disease—mechanical congestion of the sinus airspaces, infection by micro-organism (bacterial, fungal, biofilm, etc) and host inflammatory response leading to mucosal edema. We have addressed this therapeutic challenge by combining irrigation, antiseptics and anti-inflammatories. This strategy can address the underlying infectious pathology from both planktonic and biofilm sources can alleviate the concomitant inflammatory response and, mechanically clear the congested sinuses. This tripartite approach to chronic sinus disease addresses all causative pathways and chemically reduces the host immune response that leads to prolonged disease.

Other Embodiments

[0091] It is to be understood that while the invention has been described in conjunction with the detailed description thereof, the foregoing description is intended to illustrate and not limit the scope of the invention, which is defined by the scope of the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims. All publications referenced herein are incorporated by reference in their entireties.