FORMULATION

Abstract

A composition, for example an oil-in-water emulsion, comprising an amphoteric surfactant, alkoxylated cetyl alcohol, a polar dmg and a preservative, wherein the total amount of preservative present in the composition is about 0.2% w/w or less and is not benzyl alcohol and/or triclosan. The drug may be sodium cromoglicate and/or nedocromil sodium. The formulation may be useful in the treatment of skin diseases such as atopic dermatitis.

Claims

1. A composition comprising an amphoteric surfactant, an alkoxylated cetyl alcohol, a polar drug and a preservative, wherein the total amount of preservative present in the composition is about 0.2% w/w or less and is not benzyl alcohol and/or triclosan.

2. A composition according to claim 1, wherein the total amount of preservative is from about 0.001% to about 0.2% w/w of the composition, preferably from about 0.01% to about 0.15% w/w, still more preferably about 0.1% of the composition.

3. A composition according to any one of the preceding claims, wherein the preservative is chlorocresol.

4. A composition according to any one of the preceding claims, wherein the polar drug is an anionic drug.

5. A composition according to any one of the preceding claims, wherein the amphoteric surfactant is a balanced amphoteric surfactant.

6. A composition according to any of the preceding claims, wherein the alkoxylated cetyl alcohol is polypropoxylated cetyl alcohol.

7. A composition according to any of the preceding claims, wherein the amphoteric surfactant comprises disodium cocoamphodiacetate.

8. A composition according to any of the preceding claims, wherein the polar drug comprises sodium cromoglicate and/or nedocromil sodium.

9. A composition according to any one of claims 1 to 7, wherein the polar drug comprises a corticosteroid and/or an antibacterial agent.

10. A composition according to any one of claims 1 to 7, wherein the drug comprises an antirheumatic agent, nicotine or a hormone.

11. A composition according to any one of the preceding claims further comprising a corticosteroid.

12. A composition according to any one of the preceding claims, wherein the composition comprises an aqueous phase and an oil phase.

13. A composition according to claim 12 wherein the composition is an oil-in-water emulsion.

14. A composition according to any one of the preceding claims, wherein the composition is a foam, a cream, an ointment or a lotion.

15. A composition according to any one of the preceding claims, wherein the composition is for topical administration.

16. A composition according any one of claims 1 to 15 packaged in a tube, tub, bottle or pressurised aerosol container.

17. A composition according to any one of claims 1 to 16 for use in medicine.

18. A composition according to any one of claims 1 to 16 for use in the treatment of a skin disease or condition

19. A composition for use according to claim 18, wherein the skin disease or condition is a skin disease or condition involving skin mast cells and/or delayed (cellular) hypersensitivity reactions and/or inflammation.

20. A composition for use according to claim 18 or 19, in which the disease or condition is selected from atopic dermatitis, eczema, contact sensitivity, psoriasis, drug sensitivity reactions, apthous ulcers, Behcet's syndrome, pemphigus, urticaria, urticaria pigmentosa, pyroderma gangrenosum, chronic skin ulcers, skin ulcers associated with Crohn's disease, burns, insect stings/bites, herpetic infections, systemic sclerosis (systemic scleroderma), morphoea (circumscribed or localised scleroderma), dermal nodular fibrosis and sunburn.

21. A composition for use according to any one of claims 18 to 20 for use in combination with a corticosteroid.

22. A composition according to any one of claims 1 to 16 for use in the treatment of a patient in need of a polar drug.

23. A composition for use according to claim 22, wherein the patient is a patient with arthritis and wherein the polar drug is a polar anti-inflammatory or antirheumatic agent.

24. A composition for use according to claim 22, wherein the patient has acne and wherein the polar drug is a polar antibacterial drug.

25. A composition for use according to claim 22, wherein the patient is in need of nicotine and wherein the polar drug is nicotine,

26. A composition for use according to claim 22, wherein the patient is in need of nicotine and wherein the polar drug is sodium cromoglicate andior nedocromil sodium.

27. A composition according to any of claims 1 to 16 or composition for use according to any of claims 17 to 26, wherein the alkoxylated cetyl alcohol is polypropoxylated cetyl alcohol.

Description

BRIEF DESCRIPTION OF THE FIGURES

[0119] FIG. 1: Graph showing a comparison between a 4% sodium cromogycate composition of the invention and a reference composition using Tuffryn Polysulfone Membrane.

[0120] FIG. 2: Graph showing a comparison between a 4% sodium cromogycate composition of the invention and a reference composition using PVDF (hydrophilic) Membrane.

[0121] FIG. 3: Graph showing a comparison between a 4% sodium cromogycate composition of the invention and a reference composition using PVDF (hydrophobic) Membrane.

[0122] FIG. 4: Graph showing a comparison between a 4% sodium cromogycate composition of the invention and a reference composition using EpiDerm Membrane.

EXAMPLE 1

Preparation of a Composition of the Invention in the Form of an Oil-In-Water Emulsion Comprising Sodium Cromoglicate

[0123] The following substances are combined to form an emulsion. The percentages refer to percentages w/w of the final emulsion.

[0124] Group A

TABLE-US-00002 Emulsifying wax 7.2% w/w Liquid paraffin 4.8% w/w White soft paraffin  12% w/w

[0125] Group B

TABLE-US-00003 Sodium cromoglicate 4% w/w Chlorocresol 0.11% w/w Disodium cocoamphodiacetate 0.92% w/w Sodium lauryl sulfate 0.26% w/w Hexylene glycol 0.14% w/w Sodium chloride 0.06% w/w Procetyl AWS# 1% w/w Citric acid monohydrate 0.1% w/w Purified water 69.41% w/w

[0126] The emulsion was generally prepared by heating the oils (for example, the compounds in group A) to about 70° C., then adding them steadily to the water phase (for example, the compounds in Group B; also at or about 70° C.) with good stirring, and then allowing the emulsion to cool. Batches of about 10 litres to about 500 litres or more were prepared. The emulsion was prepared using a high shear homogeniser, as known to those skilled in the art. The mixture was stirred as the ingredients were mixed and stirring continued until the mixture cooled to room temperature.

[0127] An example of a specific method is as described below:

[0128] Approximately 90% of the purified water was placed into a suitable vessel for heating. Sodium cromoglicate was added to the water and heated until dissolved. Chlorocresol was then added to the water and mixed until dissolved. The aqueous mixture was then placed into a bulk manufacturing vessel. In a separate vessel some of the remaining purified water was added to sodium chloride and mixed until the sodium chloride was dissolved. Sodium lauryl sulfate, hexylene glycol and disodium cocoamphodiacetate were added to the aqueous sodium chloride solution and mixed until dissolved (this mixture is now known as Altocrom surfactant blend). White soft paraffin, liquid paraffin, emulsifying wax. The Altocrom surfactant blend and Procetyl AWS were then mixed together and heated. Meanwhile, citric acid was dissolved in the remaining purified water. Both mixtures were then added to the bulk manufacturing vessel and homogenized. The resulting composition was then cooled.

EXAMPLE 2

Preservative Optimisation

[0129] Nine compositions were prepared substantially as defined in Example 1, except for different preservatives and different amounts of preservatives were present in the compositions as shown in Table 1 below.

[0130] The microbiological stability was of each formulation was challenged using the method of the European Pharmacopoeia.

TABLE-US-00004 TABLE 1 Comparison of the microbiological stability of compositions using different preservatives and different preservative amounts. Sample Amount and Type of Preservative % w/w Number Triclosan Benzyl Alcohol Chlorocresol Result 1 0.2 — — Fail 2 — — 0.1 Pass 3 — 0.2 — Fail 4 0.2 0.2 — Fail 5 0.2 — 0.1 Pass 6 — 0.2 0.1 Pass 7 0.2 0.2 0.2 Pass 8  0.18  0.18  0.09 Pass 9 0.2 0.2 0.1 Pass

[0131] The result show that it is possible to reduce the concentration of the preservative present in the composition to 0.1% w/w and eliminate the use of triclosan and benzyl alcohol from the composition, with compositions within the scope of the invention showing a microbiological pass using chlorocresol alone at 0.1% w/w.

[0132] This result is unexpected and advantageous as the use of benzyl alcohol and/or triclosan, have the potential to cause irritation, sensitivity or allergic reaction when applied to the skin. Therefore, their use should be avoided or reduced to a minimum.

EXAMPLE 3

Determination of the Lowest Effective Level of Preservative

[0133] Five Altocrom compositions were prepared as defined in Example 1, except that the amount of chlorocresol preservative in the composition was varied as shown in Table 2 below.

[0134] The microbiological stability was of each formulation was challenged using the method of the European Pharmacopoeia.

TABLE-US-00005 TABLE 2 Evaluation of lowest level of preservative. Amount of Preservative in Composition Test Result 0.11% Pass 0.08% Pass

[0135] EXAMPLE 4

Bioavailability of a Composition of the Invention as Prepared in Example 1

[0136] The bioavailability of the composition of the invention was first investigated using a vertical diffusion cell test system.

[0137] Details of Vertical Diffusion Cell Test System

[0138] The HDT 1000 Vertical Diffusion Cell Test System was used for the studies. This system accommodates 10 diffusion cells which are situated in a heated aluminium block with a magnetic stirrer positioned under each cell.

[0139] Cells

[0140] The cell is a Type “B” cell as described in Chapter <1724>of the current US Pharmacopeia USP24—NF19 and has an aperture of 1 cm.sup.2 and a volume of ˜7 ml without the presence of the magnetic stirrer bar. The cell is made of inert borosilicate glass and fitted with a side sampling arm.

[0141] The cells contents were continuously stirred during the operation by the magnetic stirrer bar to ensure homogenous distribution of temperature and adequate mixing of contents. An “open” cell top was used for the studies.

[0142] Vacuum Deaeration Apparatus for Receptor Medium Preparation

[0143] One of the main problems with Vertical Cell Diffusion Testing is the accumulation of air bubbles on the underside of the membrane, therefore the apparatus was degassed prior to use.

[0144] The receptor media was degassed by heating it to 45° C. under a vacuum of −90 kPa whilst stirring. This system produced media with oxygen levels below 4 ppm.

[0145] Altocrom 4% Lotion Samples

[0146] Materials

[0147] Altocrom 4% Lotion made using the method described in Example 1.

[0148] Membranes

[0149] The synthetic membranes used in this study were as follows:

[0150] HT Tuffryn Polysulfone Membranes

[0151] The Tuffryn Polysulfone membrane is a relatively inert hydrophilic polymeric membrane with a pore size of 0.45 μm. Manufactured by PALL. https://www.sigmaaldrich.com/catalog/product/aldrich/z269271!land=en&region=GB

[0152] PVDF Membranes

[0153] PVDF Membrane Cat No HVLP02500 is a hydrophilic polymeric membrane with a pore size of 0.45 μm. Manufactured by Merck. http://www.merckmillipore.com/GB/en/product/Duropore-Membrane-Filter-0.4mMm NF-HVLP02500 PVDF Membrane Cat No. HVHP02500 is a hydrophobic membrane with a pore size of 0.45 μm. http://www.merckmillipore.com/GB/en/product/Duropore-Membrane-Filter-0.45m,MM NF-HVHP02500

[0154] EpiDerm

[0155] EpiDerm membranes are Reconstructed Human Epidermis (RHE). They are composed of normal, human-derived epidermal keratinocytes (NHEK) cultured on specially prepared tissue culture inserts.

[0156] EpiDerm has a human epidermal tissue structure and cellular morphology. It has a 3D structure consisting of organised and proliferative basal cells, spinous and granular layers and cornified epidermal layers.

[0157] EpiDerm Part Numer EPI-606 was used for this study and was supplied by MatTek Corporation. Supplier's storage and preparation instructions were followed to prepare the EpiDerm prior to the testing. https://www.mattel.com/product-category/tissure-models/epiderm/

[0158] Receptor Medium

[0159] Phosphate buffered saline (PBS) solution containing streptomycin (0.1 mg/ml) and penicillin (100 units/ml) was used as the receptor fluid. This was prepared by adding five tablets of PBS with Penicillin-Streptomycin (10 ml) and making up to 1 l. This was degassed as described above.

[0160] Vertical Diffusion Cell Preparation

[0161] The Vertical Diffusion Cells were prepared by firstly adding a stirrer bar and then filling with degassed receptor medium (Phosphate Buffered Saline (PBS) Solution containing streptomycin (0.1 mg/ml) and penicillin (100 units/ml) as described above). The cell body was filled with the media to give a positive meniscus at the top of the cell to reduce any air being trapped under the membrane.

[0162] The assembled sample holder (open top) with the selected membrane was then placed on top of the cell body with the membrane contacting the receptor media.

[0163] The cell was checked for any air bubbles and then Parafilm wrapped around the joint between the cell top and body.

[0164] The prepared cell was placed in the heating block whilst the other cells were being prepared to maintain the required temperature of 32° C.

[0165] When all the cells had been filled with receptor media, the cell was removed with the holder and placed on a 4 dp balance. The Altocrom 4% Lotion was applied to the membrane with a 10 μl displacement micropipette and the exact mass recorded. The cream was then evenly distributed over the membrane with the end of a small glass rod. The time when the cream was applied was recorded as t=0. A layer of Parafilm was used to cover the open top of the cell.

[0166] For comparing the old and new formulation, 5 cells were dosed with the composition as prepared in Example 1 and 5 cells with a comparative composition.

[0167] Receptor Media Sampling

[0168] Sampling of the receptor media was carried out at the desired time points. Prior to sampling (approx. 10 min), the volume in the cell was checked against the calibration mark and refilled if required. Sampling was carried out using 100 μl displacement micropipette. 100 μl of the receptor fluid was placed in a glass HPLC vial with 300 μl insert. The receptor fluid was replaced immediately after sampling.

[0169] HPLC Analysis to Determine Sodium Cromoglicate (% w/w) in Receptor Medium

[0170] The following method describes the analysis to determine the sodium cromoglicate present in the receptor medium.

[0171] Mobile Phase: 77% Purified Water/23% Acetonitrile. Adjusted to pH 1.95-2.05 with Orthophosphoric Acid.

[0172] PBS Solution: 5 tablets of Phosphate Buffered Saline and 10.0 ml of Penicillin—Streptomycin made up to 1 l with purified water.

[0173] Standard Stock Solution: The Approved Reference Standard of Sodium Cromoglicate was accurately weighed to give 0.40 g±0.04 g (W.sub.1). This was added to a 100 ml volumetric flask and dissolved in purified water and made up to volume.

[0174] Calibration Standard Solution: The appropriate concentration was selected for the analysis being carried out

[0175] As an example, for a 400 ppm calibration standard solution, 10.0 ml of the standard stock solution was pipetted into a 100 ml volumetric flask and diluted to 100 ml with PBS solution. This was equivalent to 400 ppm.

[0176] This can be diluted further with PBS to obtain the appropriate concentration to calibrate at lower concentration levels.

[0177] Conditions

[0178] Column:4 μm SYNERGI Polar-RP 25-mm×4.6 mm (or chemically or physically equivalent)

[0179] Flow rate (ml/min):2.0

[0180] Detector Wavelength (nm): 325

[0181] Injection Volume (μl): 20

[0182] Oven Temperature: 30° C.

[0183] Approx. retention times (min): Sodium cromoglicate 3-5 min

[0184] Approx, Total Run Length: 15 min

[0185] Calculations for Chromeleon Software/Integrator

TABLE-US-00006 TABLE 3 Peak Table Peak Name Amount Response Factor 1 Sodium Cromoglicate (W.sub.1/0.4) × c 1.0

TABLE-US-00007 TABLE 4 Sequence (Sample List) Type Weight Dilution Factor ISTD Amount STD 1.0 1.0 1.0 SAMPLE 1.0 0.7 1.0

[0186] Results

[0187] Tuffryn Polysulfone Membrane

[0188] The procedure described in the Vertical Diffusion Cell Preparation section detailed above was carried out using the Tuffryn Polysulfone membrane and pre-wetting the membrane was carried out.

[0189] Sampling was carried out at 4 and 24 hr and the results are shown in Table 5 and FIG. 1.

TABLE-US-00008 TABLE 5 Percentage of SCG recovery % SCG recovery/cm.sup.2 of membrane Time/hr Comparative composition Composition of Example 1 0 0 0 4 60.20 ± 25.9  68.60 ± 18.21 24 65.60 ± 15.59 71.11 ± 5.20 

[0190] The results showed that both compositions provided significant levels of time dependent penetration of cromoglicate from the compositions, with up to 65% achieved after 5 hours for the comparative composition and up to 71% achieved after 5 hours for the composition of Example 1.

[0191] PVDF (Hydrophilic) Membrane

[0192] The same method was carried out as described in the Vertical Diffusion Cell Preparation section detailed above. Pre-wetting the membrane was found to leave a varying amount of PBS solution on the surface of the membrane which caused the cream to be quickly diluted by an unknown quantity and added to experimental error. The membranes were therefore used without pre-wetting but were observed to hydrate very quickly with the PBS solution in the body of the Vertical Diffusion Cell.

[0193] The SCG recovery is shown in Table 6 and FIG. 2.

TABLE-US-00009 TABLE 6 SCG recovery using PVDF membrane % SCG recovery/cm.sup.2 of membrane Time/hr Comparative composition Composition of Example 1 0 0 0 1 59.68 ± 9.84 56.40 ± 4.44 4 73.64 ± 5.80 66.12 ± 8.07

[0194] The results showed that both compositions provided significant levels of time dependent penetration of the cromoglicate from the compositions, with up to about 60% achieved after 1 hour for both compositions.

[0195] PVDF (Hydrophobic) Membranes

[0196] The same method was carried out as described in the Vertical Diffuesion Cell Preparation section detailed above. The hydrophobic nature of the membranes was found to make pre-wetting very difficult. Therefore the method was conducted without pre-wetting the membrane.

[0197] It was apparent that the percentage of the sodium cromoglicate recovered reached a maximum quickly so the samples were not continued for 24 hr.

[0198] The results are shown in Table 7 and FIG. 3.

TABLE-US-00010 TABLE 7 SCG recovery using PVDF membrane % SCG recovery/cm.sup.2 of membrane Time/hr Comparative composition Composition of Example 1 0 0 0 1 29.72 ± 13.11 37.05 ± 28.18 2 58.21 ± 10.67 63.87 ± 6.37  3 60.51 ± 6.72  65.51 ± 4.14  4 58.02 ± 11.02 56.80 ± 12.07

[0199] The results showed that both compositions provided significant levels of time dependent penetration of the cromoglicate from the compositions, with up to 60% achieved after 3 hours for the comparative composition and up to 65% achieved after 3 hours for the composition of Example 1.

[0200] EpiDerm

[0201] The membrane was stored and prepared as specified by the supplier instructions. Prior to use, the EpiDerm membranes were removed from the inserts they were supplied in using forceps and carefully transferred (with the supporting PTFE membrane underneath) and placed in pre-warmed assay medium at 37° C. The EpiDerm was incubated for 1 hr prior to use.

[0202] Sampling was carried out after 1, 2, 3, 4 and 24 hr and the results are shown in Table 8 and FIG. 4.

TABLE-US-00011 TABLE 8 SCG recovery using EpiDerm membrane % SCG recovery/cm.sup.2 of membrane Time/hr Comparative composition Composition of Example 1 0 0 0 1 0.02 ± 0.04 0.14 ± 0.13 2 0.13 ± 0.22 0.28 ± 0.24 3 0.56 ± 0.60 0.93 ± 0.68 4 0.95 ± 0.91 1.67 ± 1.14 5 1.68 ± 1.29 2.16 ± 1.25 24 6.81 ± 3.30 8.11 ± 3.42

[0203] The results showed that both compositions provided some time dependent penetration of the cromoglicate from the compositions, with up to 6% achieved after 24 hours for the comparative composition and up to 8% achieved after 24 hours for the composition of Example 1.

CONCLUSIONS

[0204] The tests clearly demonstrate that the composition of Example 1 (a composition of the invention), provides good penetration/absorption of sodium cromoglicate on multiple different membranes achieving levels that were typically better than the comparative composition.

[0205] The high level of absorption shown by the compositions of the invention would be expected to be further enhanced when used to treat conditions such as atopic dermatitis because the in vitro tests do not take account of the effect that rubbing the product into the skin, which is known to aid splitting of the composition into water and oil phases, occluding the cromoglicate in the aqueous phase and therefore aiding absorption. Also, in atopic dermatitis the skin barrier is compromised hence further aiding penetration.

EXAMPLE 5

Long Term Stability of a Composition of the Invention

[0206] An example of the composition of the invention was prepared as described in Example 1.

[0207] The composition of the invention comprises an amphoteric surfactant an alkoxylated cetyl alcohol and a polar drug, wherein the total amount of preservative present in the composition is about 0.2% wlw or less of the composition and is prepared as detailed in Example 1. The condition that the compositions were tested under are summarised in Table 9.

[0208] Batches subjected to long term stability testing were as follows.

TABLE-US-00012 TABLE 9 Test condition details of a composition of the invention comprising about 0.2% w/w or less of at least one preservative. Batch Number Orientation Conditions 1 Upright 25° C./60% RH 30° C./65% RH 40° C./75% RH 2 Upright 25° C./60% RH 30° C./65% RH 40° C./75% RH

[0209] The stability specification that the compositions were assessed under is presented in Table 10.

TABLE-US-00013 TABLE 10 Stability specification (House refers to the in-house standard and Ph. Eur refers to the standard set out in the European Pharmacopoeia). Attribute Specification Monograph Method/Comment Appearance A smooth, white lotion free House Visual from particulate contamination. Product is uniform and has not separated Identification: UV/ The ratio of absorbance at House UV/VIS Sodium Visible 239 nm and 327 nm is between Not routinely tested Cromoglicate 0.25-0.30 Chemical/ A yellow/green colour is House Chemical/ colourmetric produced colourmetric. Not routinely tested HPLC The peak in the sample House HPLC chromatogram has the same Not routinely tested retention time as the peak in the standard chromatogram Assay of Sodium 3.8-4.2 House HPLC Cromoglicate (% w/w) Chlorocresol (% w/w) 0.10-0.12 (release) House HPLC 0.08-0.12 (shelf life) pH at 20° C. 3.8-5.8 House pH meter Penetration at 25° C. 100-300 House Penetrometer (tenths mm) *Kromo I (% w/w relative Not greater than 0.20 House HPLC to Sodium Cromoglicate) *Kromo II (% w/w relative Not greater than 0.20 House HPLC to Sodium Cromoglicate) *Oxalate (% w/w relative to Not more than 0.10 House Ion Sodium Cromoglicate) Chromatography Any individual unknown Not more than 0.10 House HPLC degradant (% w/w relative to sodium Cromoglicate) *Total specified Not more than 0.50 House HPLC/Ion degradants (% w/w relative Chromatography to Sodium Cromoglicate) Total degradation products Not more than 1.0 House HPLC/Ion (% w/w relative to Sodium Chromatography Cromoglicate) Total Aerobic Microbial Not more than 10.sup.2 aerobic Ph. Eur. Ph. Eur. Cotint organisms/gram (colony forming units per gram) Total Yeasts & Mould Not more than 10.sup.3 yeasts & Ph. Eur. Ph. Eur. Count mould/gram (colony forming units per gram) Absence of Absent in 1 gm Ph. Eur. Ph. Eur. Enterobacteriaceae Absence of Pseudomonas Absent in 1 gm Ph. Eur. Ph. Eur. aeruginosa Absence of Absent in 1 gm Ph. Eur. Ph. Eur. Staphylococcus aureus Preservative Efficacy Ph. Complies Ph. Eur. Ph. Eur. Eur. Topical preparations- Performed only on Criteria A batch 2359 *no longer tested

[0210] Summary of Stability Data on the Composition of the Invention

[0211] A summary of stability data on the composition of the invention as prepared in Example 1. The results are shown in Table 11.

[0212] Appearance

[0213] Remains in specification throughout 36 months testing.

[0214] Assay of Sodium Cromoglicate (% w/w)

[0215] Remains in specification throughout 36 months testing. No trend apparent at any storage condition.

[0216] Chlorocresol (% w/w)

[0217] This attribute trends downwards significantly at accelerated conditions. The initial shelf life specification of 0.10-0.12% w/w was reset to 0.08-0.12% w/w on the basis of stability test results (preservative efficacy testing having established that this level provided adequate preservation). All batches remained in specification to 36 months at this condition.

[0218] pH at 20° C.

[0219] Remains in specification throughout 36 months testing. No trend apparent at any storage condition.

[0220] Penetration at 25° C.

[0221] Remains in specification throughout 36 months testing.

[0222] Kromo I (% w/w relative to Sodium Cromoglicate)

[0223] Remains in specification throughout 36 months testing. No trend apparent at any storage condition.

[0224] Kromo II (% w/w relative to Sodium Cromoglicate)

[0225] Remains in specification throughout 36 months testing. No trend apparent at any storage condition.

[0226] Oxalate (% w/w relative to Sodium Cromoglicate)

[0227] Remains in specification throughout 36 months testing. No trend apparent at any storage condition.

TABLE-US-00014 TABLE 11 Results of Stability Tests Long term stability testing - upright storage Batches DEV2359, DEV2360 25° C./60% RH 30° C./65% RH 40° C./75% RH Attribute Specification to 36 months to 36 months to 36 months Appearance Smooth, white lotion Complies Complies Complies free from particulate contamination Assay of Sodium Cromoglicate (% w/w) 3.8-4.2 3.8-4.1 3.8-4.1 3.9-4.2 Chlorocresol (% w/w) 0.08-0.12 0.10-0.11 0.10-0.11 0.06-0.11 pH at 20° C. 3.8-5.8 4.7-5.0 4.1-4.9 4.7-5.0 Penetration at 25° C. (tenths min) 100-300 105-215 121-227 115-234 Kromo I (% w/w relative to Sodium Not greater than 0.20 None detected None detected None detected Cromoglicate) Kromo II (% w/w relative to Sodium Not greater than 0.20 None detected None detected None detected Cromoglicate) Oxalate relative to Sodium Not more than 0.10 0.035-0.046 0.035-0.047 0.036-0.049 Cromoglicate) Any individual unknown degradant Not more than 0.10 None detected None detected None detected (% w/w relative to Sodium Cromoglicate) Total specified degradants (% w/w Not more than 0.50 0.033-0.045 0.035-0.047 0.036-0.049 relative to Sodium Cromoglicate) Total degradation products (% w/w Not more than 1.0 0.033-0.045 0.035-0.047 0.036-0.049 relative to Sodium Cromoglicate) Total Viable Count Not more than 10.sup.2 Complies Complies — aerobic microorganisms per gram (colony forming units per gram) Absence of Enterobacteriaceae Absent in 1 gm Complies Complies — Absence of Pseudomonas aeruginosa Absent in 1 gm Complies Complies — Absence of Staphylococcus aureus Absent in 1 gm Complies Complies — Preservative Efficacy Ph. Ear. Complies Pass Pass — Topical preparations- Criteria A

[0228] Any individual unknown degradant (% w/w relative to Sodium Cromoglicate)

[0229] Remains in specification throughout 36 months testing. No trend apparent at any storage condition.

[0230] Total specified degradants (% w/w relative to Sodium Cromoglicate)

[0231] Remains in specification throughout 36 months testing. No trend apparent at any storage condition.

[0232] Total degradation products (% w/ane relative to Sodium Cromoglicate)

[0233] Remains in specification throughout 36 months testing. No trend apparent at any storage condition,

[0234] Total Viable Count

[0235] Remains in specification throughout 36 months (testing at 25° C./60% RH and 30° C./65% RH only).

[0236] Absence of Enterobacteriaceae, Pseudomonas aeruginosa and Staphylococcus aureus

[0237] Remains in specification throughout 36 months (testing at 25° C./60% RH and 30° C./65% R only).

[0238] Preservative Efficacy Ph. Eur. Topical preparations—Criteria A

[0239] Performed only on batch 2359, Remains in specification throughout 36 months (testing at 25° C./60% RH and 30° C./65% RH only).

[0240] Freeze-Thaw Cycling

[0241] Freeze-Thaw cycling has not been performed on this product. It is not considered necessary—it is well established that emulsions of this type are not physically stable and will crack when frozen.

[0242] In Use Stability Tests

[0243] In-use stability testing was performed to replicate use over 56 days at 25° C./60% RH. The testing included two aged batches (24 and 36 months) of the composition of the invention prepared as defined in Example 1. All parameters tested remained in specification at 56 days all both batches tested. The results are presented in Tables 12, 13 and 14 below.

TABLE-US-00015 TABLE 12 Stability of a composition of the invention in use over 56 at 25° C./60% RH in a 150 ml plinth HDPE white round bottle with Model 7 pump dispenser. Attribute Specification Initial 56 Days Appearance Smooth, white lotion free Complies Complies from particulate Assay of Sodium Cromoglicate (% w/w) 3.8-4.2 4.0 3.9 Chlorocresol (% w/w) 0.10-0.12 0.11 0.11 pH at 20° C. 3.8-5.8 4.8 4.7 Kromo I (% w/w relative to Sodium Not greater than 0.20 None None Cromoglicate) detected detected Kromo II (% w/w relative to Sodium Not greater than 0.20 None None Cromoglicate) detected detected Oxalate relative to Sodium Not more than 0.10 0.037 0.040 Cromoglicate) Any individual unknown degradant Not more than 0.10 None None (% w/w relative to Sodium detected detected Cromoglicate) Total specified degradants (% w/w Not more than 0.50 0.037 0.040 relative to Sodium Cromoglicate) Total degradation products (% w/w Not more than 1.0 0.037 0.040 relative to Sodium Cromoglicate) *Total Viable Count Not more than 10.sup.2 Complies Complies aerobic organisms/gram Absence of Enterobacteriaceae Absent in 1 gm Complies Complies Absence of Pseudomonas aeruginosa Absent in 1 gm Complies Complies Absence of Staphylococcus aureus Absent in 1 gm Complies Complies Preservative Efficacy Ph. Eur. Complies Complies Complies Topical preparations- Criteria A *Weekly microbiological testing was also passed at each time point

TABLE-US-00016 TABLE 13 Stability of a composition of the invention aged for 24 months in use over 56 at 25° C./60% RH in a 150 ml plinth HDPE white round bottle with Model 7 pump dispenser. Attribute Specification Initial 56 Days Appearance Smooth, white lotion free Complies Complies from particulate Assay of Sodium Cromoglicate (% w/w) 3.8-4.2 4.1 3.9 Chlorocresol (% w/w) 0.10-0.12 0.10 0.10 pH at 20° C. 3.8-5.8 4.8 4.9 Penetration at 25° C. (tenths min) 100-300 140 157 Kromo I (% w/w relative to Sodium Not greater than 0.20 None None Cromoglicate) detected detected Kromo II (% w/w relative to Sodium Not greater than 0.20 None None Cromoglicate) detected detected Oxalate relative to Sodium Not more than 0.10 0.042 0.043 Cromoglicate) Any individual unknown degradant Not more than 0.10 None None (% w/w relative to Sodium detected detected Cromoglicate) Total specified degradants (% w/w Not more than 0.50 0.042 0043 relative to Sodium Cromoglicate) Total degradation products (% w/w Not more than 1.0 0.042 0.043 relative to Sodium Cromoglicate) Total Viable Count Not more than 10.sup.2 Complies Complies aerobic organisms/gram Absence of Enterobacteriaceae Absent is 1 gm Complies Complies Absence of Pseudomonas aeruginosa Absent is 1 gm Complies Complies Absence of Staphylococcus aureus Absent is 1 gm Complies Complies Preservative Efficacy Ph. Eur. Complies — Complies Topical preparations- Criteria A

TABLE-US-00017 TABLE 14 Stability of a composition of the invention aged for 36 months in use over 56 at 25° C./60% RH in a 150 ml plinth HDPE white round bottle with Model 7 pump dispenser. Attribute Specification Initial 56 Days Appearance Smooth, white lotion free Complies Complies from particulate Assay of Sodium Cromoglicate (% w/w) 3.8-4.2 3.9 4.0 Chlorocresol (% w/w) 0.10-0.12 0.11 0.11 pH at 20° C. 3.8-5.8 5.0 4.8 Penetration at 25° C. (tenths min) 100-300 148 193 Kromo I (% w/w relative to Sodium Not greater than 0.20 None None Cromoglicate) detected detected Kromo II (% w/w relative to Sodium Not greater than 0.20 None None Cromoglicate) detected detected Oxalate relative to Sodium Not more than 0.10 0.042 0.039 Cromoglicate) Any individual unknown degradant Not more than 0.10 None None (% w/w relative to Sodium detected detected Cromoglicate) Total specified degradants (% w/w Not more than 0.50 0.042 0.039 relative to Sodium Cromoglicate) Total degradation products (% w/w Not more than 1.0 0.042 0.039 relative to Sodium Cromoglicate) Total Viable Count Not more than 10.sup.2 Complies Complies aerobic organisms/gram Absence of Enterobacteriaceae Absent to 1 gm Complies Complies Absence of Pseudomonas aeruginosa Absent to 1 gm Complies Complies Absence of Staphylococcus aureus Absent to 1 gm Complies Complies Preservative Efficacy Ph. Ear. Complies Pass Pass Topical preparations- Criteria A *Weekly microbiological testing was also passed at each time point

[0244] Forced Degradation

[0245] Forced degradation studies of the product with add, base and peroxide were performed as a screen for potential degradation products. The samples were evaluated by LCMSMS.

[0246] The samples were mixed with acid, base or peroxide and left overnight. The samples were then diluted with water and filtered.

[0247] The samples were then analysed using LCMSMS. The samples were run on an Agilent 1290 with Agilent 6530 qTOF mass spectrometry.

[0248] The chromatograms were compared and any evidence of new peaks when compared to the relevant controls were recorded and their mass determined.

[0249] Significant degradation was seen only in alkaline solution. The structure of cromoglicate is such assignment of structures is difficult, The molecular formulas of theses degradation peaks have been calculated but they cannot be assigned to an individual structure. As sodium cromoglicate is well known, and as Altocrom in use will not be exposed to strongly alkaline conditions, the inability to assign structures is not considered important.

[0250] Proposed Shelf Life

[0251] The proposed shelf life is 36 months when the product is stored at a temperature not exceeding 25° C.