CA2391976A1 - Use of fatty acid/amino acid soaps to enhance antimicrobial effectiveness of topical pharmaceutical compositions - Google Patents

Abstract

Fatty acid/amino acid soaps are used to enhance antimicrobial effectiveness in topically administrable pharmaceutical compositions containing at least one active ingredient, a cationic preservative and an anionic polyelectrolyte, such as a carboxyvinyl polymer, xanthan gum, polystyrene sulfonic acid polymer or cationic exchange resin.

Description

USE OF FATTY ACID/ AMINO ACID SOAPS TO ENHANCE
ANTIMICROBIAL EFFECTIVENESS OF TROPICAL
PHARMACEUTICAL COMPOSITIONS
s This application claims priority to co-pending U.S. Provisional Application, U.S. Serial No. 60/105,855, filed October 27, 1998.
Background of the Invention ,o The present invention relates generally to the preservation of pharmaceutical compositions. In particular, the present invention relates to the use of fatty acid/amino acid soaps to prevent or to reduce binding of the antimicrobial components of topically administrable pharmaceutical ,5 compositions to other components contained therein, thereby improving the antimicrobial efficacy of such compositions.
In recent years, a number of ophthalmic compositions have been introduced that contain a variety of components, such as carboxyvinyl Zo polymers (e.g., Carbopol~), ion exchange resins (e.g., Amberlite~), or other large polyelectrolytes, which provide sustained release of the ophthalmic agent(s), as well as increased patient comfort. Such compositions are described, for example, in US 4,911,920 (Jani et al.). Although these compositions are comfortable and have sustained release characteristics, z5 cationic antimicrobials, such as benzalkonium chloride (BAC), which are often added as preservatives to such compositions, tend to bind to the anionic polyelectrolytes present in the formulations, resulting in loss of antimicrobial effectiveness.
3o Sarcosinate surfactants are composed of acylated sarcosines.
Sarcosine (CH3-NH-CH2-COOH), an amino acid normally found in starfish and sea urchins, is chemically related to glycine (NH2-CH2-COOH), a basic amino acid in mammals. Common fatty acids and their derivatives utilized in the manufacture of sarcosinate surfactants are lauric, oleic, and myristic acids and their esters and halides. Because of their mildness, sarcosinate surfactants have been utilized in shampoos, mouthwashes, skin cleansers, sunscreens, aerosol shaving lathers and other personal care products. To date, the main applications of these types of surfactants have been in the cosmetic industry. For example, European Patent Application No. 0 194 097 (Schmidt et al.), assigned to Procter & Gamble, mentions sodium lauroyl sarcosinate as the mild anionic surfactant utilized in an aerosol skin-cleansing and moisturizer mousse.
,0 U.S. Patent No. 5,520,920 (Castillo, et al.) discloses the use of certain modified sarcosinates and lactylates to enhance antimicrobial effectiveness of ophthalmic compositions, particularly in the case where cationic preservatives otherwise bind to anionic polyelectrolytes. The modified sarcosinates have ,5 the formula:
CH3 ~O
R'~C-N-CH2-C\\ M+
O
n wherein: R~ = C4-C27 saturated or unsaturated hydrocarbon;
M = H or a pharmaceutically acceptable salt; and n=1,2or3.
Representative modified sarcosinates include those sold under the Hamposyl~ trade name, such as lauroyl sarcosine (Hamposyl° L), oleoyl sarcosine (Hamposyl° O), myristoyl sarcosine (Hamposyl~ M), cocoyl sarcosine (Hamposyl° C), stearoyl sarcosine (Hamposyl° S), and pelargodoyl sarcosine (Hamposyl~ P). Representative lactylates include sodium capryl lactylate (Pationic° 122A).

W~ 01/21209 CA 02391976 2002-02-26 PCT/US00/24765 Additional solutions to the problem of cationic preservative - anionic polyelectrolyte binding problem in topically administrable pharmaceutical compositions are desirable.
Fatty acid/amino acid soaps are known and include, for example, those surfactants sold under the Aminosoap° trade name (Ajinomoto Co., Inc., Tokyo, Japan). According to product brochures, Aminosoap~ surfactants are used in hair and body care (hair shampoo, body wash and bath foam), facial ,0 care (facial cleanser, facial washing foam, facial washing creme, and make-up remover), and household and health care.
Summary of the Invention ,5 The present invention provides a method of enhancing the preservative efficacy of topically administrable pharmaceutical compositions containing an anionic polyelectrolyte and a cationic preservative. According to the method of the present invention, a fatty acid/amino acid soap is added to the topically administrable pharmaceutical composition.
z0 Although the Applicants do not wish to be bound to a particular theory, it is believed that the addition of fatty acid/amino acid soaps to the compositions results in the release of the bound cationic preservative from the anionic polyelctrolyte by the formation of a loose and reversible 25 surfactant-preservative complex. The surfactant-preservative complex has antimicrobial effectiveness. Alternatively, the soaps may themselves possess antimicrobial activity.
Regardless of the mechanism, the fatty acid/amino acid soaps of the 30 present invention improve the preservative efficacy of topically administrable pharmaceutical compositions. Accordingly, the present invention also relates to topically administrable pharmaceutical compositions containing one or more pharmaceutically active agents, an anionic polyelectrolyte, a cationic preservative, and a fatty acid/amino acid soap.
Detailed Description of the Invention Unless indicated otherwise, all amounts of composition ingredients expressed in percentage terms are expressed as weight/weight.
Fatty acid/amino acid soaps useful in the preservative systems of the ,o present invention are those wherein the fatty acid component is derived from a C$ - C24 fatty acid and the amino acid component is selected from the group consisting of lysine and arginine. Preferably, the fatty acid component is selected from the group consisting of cocoyl, linoleoyl, lauroyl, myristoyl, stearoyl, oleoyl, and pelargodoyl fatty acid residues. Such fatty acid/amino ,s acid soaps are commercially available or can be made by known methods.
For example, the soap where the fatty acid component is cocoyl and the amino acid component is derived from arginine is commercially available as AMINOSOAP AR-12 from Ajinomoto Co., Inc. (Tokyo, Japan). The soap where the fatty acid component is cocoyl and the amino acid component is zo derived from lysine is available as AMINOSOAP LYC-12.
In general, the amount of fatty acid/amino acid soap present in the compositions of the present invention is from about 0.001 to about 1 %, preferably from about 0.01 to about 0.2%, and most preferably from about 25 0.03 to about 0.12%. For topical ophthalmic preparations, the concentration of the fatty acid/amino acid soap should not be so high that it causes severe discomfort.
The compositions of the present invention contain cationic 3o antimicrobials and anionic polyelectrolytes. Cationic antimicrobials are known in the art and include, but are not limited to, quaternary ammonium compounds, such as benzalkonium chloride and polyquaternium-1. Anionic polyelectrolytes include high molecular weight, anionic mucomimetic polymers (e.g., carboxyvinyl polymers such as Carbopol°), polystyrene sulfonic acid polymers, cationic exchange resins (e.g., Amberlite~ or Dowex°), and the like.
High molecular weight, anionic mucomimetic polymers have a molecular weight between about 50,000 and 6 million. The polymers are characterized as having carboxylic acid functional groups and preferably contain between 2 and 7 carbon atoms per functional group. Suitable high molecular weight, anionic polymers are carboxyvinyl polymers, preferably ,0 those called Carbomers, e.g., Carbopol~ (B.F. Goodrich Co., Cleveland, Ohio). Specifically preferred are Carbopol° 934P, Carbopol°
974P and Carbopol° 940. Other suitable high molecular weight, anionic polymers include: alginates, carrageenans, natural gums (xanthan, karaya and tragacanth) and carboxy methyl cellulose. Such polymers will typically be ,5 employed in an amount between about 0.05 and about 6%, depending on the desired viscosity of the composition. Pourable liquid compositions generally comprise an amount of the polymer between about 0.05 and about 2%.
Cation exchange resins are characterized as either strongly acidic, z0 such as those having sulfonic acid or sulfuric acid functionality, or weakly acidic, such as those having carboxylic acid functionality. Such resins are readily available, for example, from Rohm & Haas (Philadelphia, Pennsylvania) under the name Amberlite~ and from Dow Chemical Co.
(Midland, Michigan) under the name Dowex~. The average particle size of 25 the commercially available forms of the resins is about 40 to 150 microns.
The particle size of the resin is critical for topically administrable ophthalmic compositions. Accordingly, for topically administrable ophthalmic compositions, commercially available resin particles are reduced by known techniques, including ball milling, to a particle size of about 20 ~m or less such that the average particle size is <_ 10 Vim, and are preferably reduced to a particle size of about 10 ~m or less. Ion exchange resins are typically used in an amount from about 0.05 to about 10%.

W~ 01/21209 CA 02391976 2002-02-26 PCT/US00/24765 Anionic mucomimetic polymers and cation exchange resins are discussed in greater detail in U.S. 4,911,920 issued March 27, 1990, the entire contents of which are hereby incorporated by reference herein.
s The polystyrene sulfonic acid polymers (and their salts) useful in the compositions of the present invention comprise the following repeating unit:
W (I) ,o *~H2C- IC~
,s wherein: W = H or CH3; and x = an integer such that the molecular weight of the polystyrene sulfonic acid polymer is from about 10,000 to 1.6 million.
In the preferred polystyrene sulfonic acid polymers of formula I, W = H
and the molecular weight is between about 500,000 to about 1,000,000, preferably about 600,000. If present in the compositions of the present invention, the polystyrene sulfonic acid polymers of formula I comprise less 2s than about 8%, preferably less than about 5%.
The active ingredient or ingredients that can be included in the compositions of the present invention include all ophthalmic, dermatological, otic or nasal agents that can be topically applied. For example, such ophthalmic agents include (but are not limited to): anti-glaucoma agents, such as beta-blockers (e.g., betaxolol and timolol), muscarinics (e.g., pilocarpine), prostaglandins, carbonic anhydrase inhibitors (e.g., acetazolamide, methazolamide and ethoxzolamide), dopaminergic agonists and antagonists, and alpha adrenergic receptor agonists, such as para-amino clonidine (also known as apraclonidine) and brimonidine; anti-infectives, such as ciprofloxacin; non-steroidal and steroidal anti-inflammatories, such as suprofen, ketorolac, dexamethasone, rimexolone and tetrahydrocortisol;
proteins; growth factors, such as EGF; and anti-allergic agents, such as cromolyn sodium, emedastine and olopatadine. Compositions of the present invention may also include combinations of active ingredients. Most preferred are topically administrable ophthalmic compositions.
,o The compositions of the present invention can also include other components, for example, pharmaceutically acceptable buffers; tonicity agents; comfort-enhancing agents; solubilizing aids; pH adjusting agents;
antioxidants; and stabilizing agents. The compositions may also contain additional preservatives (in conjunction with the cationic preservatives ,5 addressed above). As will be appreciated by those skilled in the art, the compositions may be formulated in various dosage forms suitable for topical delivery, including solutions, suspensions, emulsions, and gels.
The following examples are presented to illustrate further various Zo aspects of the present invention, but are not intended to limit the scope of the invention in any respect.

Ingredient Concentration (%) Betaxolol HCI 0.28 Amberlite IRP-69 0.25 Carbopol 974P 0.45 Cocoyl N-Lysine* 0.03 Boric Acid 0.4 Mannitol 4.5 Edetate Disodium 0.01 Benzalkonium Chloride 0.01 NaOH and/or HCI q.s. to pH 6.5 Purified Water q.s. to 100 * Aminosoap LYC-12 Preparation:
0.42g betaxolol hydrochloride, 0.375g amberlite IRP 69 resin, 0.60g boric acid, 6.75g mannitol, 0.015g disodium EDTA were combined in 40mL water and stirred for 30 minutes. To this was added 1.06g 1 % BAC stock solution, 33.8g of 2% carbopol 974P stock slurry, and 0.15g of a 30% solution of ,o cocoyl lysine (Aminosoap LYC-12 from Ajinomoto.) The pH of the formulation was adjusted to 6.5 by the addition of 2.2mL of 5N NaOH and the final batch amount was brought to 150mL with water. The formulation was sterilized for 60 minutes in an autoclave oven at 121 °C.

Ingredient Concentration (%) Betaxolol HCI 0.28 Amberlite IRP-69 0.25 Carbopol 974P 0.45 Cocoyl N-Lysine* 0.03 Boric Acid 0.4 Mannitol 4.5 Edetate Disodium 0.01 Benzalkonium Chloride 0.01 + 10%xs Tromethamine q.s. to pH 6.5 Purified Water q.s. to 100 *Aminosoap LYC-12 Preparation:
0.84g betaxolol hydrochloride and 0.75g amberlite IRP 69 resin were combined in ~30 mL water and stirred for 30 minutes. To this was added (in order indicated) 67.5g of 2% carbopol 974P stock slurry, 13.5g mannitol, 0.03g disodium EDTA, 1.20g boric acid, and 2.97g of 1.11 % benzalkonium ,o chloride stock solution. To this was added 25mL of 10% tromethamine solution, and the mixture was allowed to stir for 2 hours. The formulation was brought to 260g by the addition of water, and the pH was adjusted to 6.0 by the dropwise addition of 10% tromethamine solution. To the formulation was added 0.3g of 30% cocoyl lysine solution (Aminosoap LYC-12 from ,5 Ajinomoto). The formulation was adjusted to pH 6.5 by the dropwise addition of 10% tromethamine solution, and the final batch amount was then adjusted to 300g by the addition of water. The formulation was steam sterilized for 60 minutes at 121°C in an autoclave oven.

The formulation of Example 2 was found to have the following characteristics.
label benzalkonium chloride 110% (prepared to contain 10% excess) unbound benzalkonium chloride 0.76%
label betaxolol 100%
unbound betaxolol 28%
osmolality 312 mOsm/Kg viscosity 138 cps ,o EXAMPLE 3 Re presentative Formulation Ingredient Concentration (%) Betaxolol HCI 0.28 Amberlite IRP-69 0.25 Carbopol 974P 0.45 Cocoyl N-Lysine* 0.06 Boric Acid 0.4 Mannitol 4.5 Edetate Disodium 0.01 Benzalkonium Chloride 0.01 NaOH and/or HCI q.s. to pH 7.2 Purified Water q.s. to 100 * Aminosoap LYC-12 WO 01/21209 CA 02391976 2002-02-26 pCT/LTS00/247G5 Ingredient Concentration (%) Betaxolol HCI 0.28 Amberlite IRP-69 0.25 Carbopol 974P 0.45 Cocoyl N-Arginine* 0.03 Boric Acid 0.4 Mannitol 4.15 Edetate Disodium 0.01 Benzalkonium Chloride 0.01 + 10%xs Tromethamine q.s. to pH 6.5 Purified Water q.s. to 100 * Aminosoap AR-12 Representative Formulation Ingredient Concentration (%) Betaxolol HCI 0.28 Brinzolamide 1 Amberlite IRP-69 0.25 Carbopol 974 P 0.45 Boric Acid 0.4 Mannitol 4.5 Cocoyl N-Lysine* 0.06 Edetate Disodium 0.01 Benzalkonium Chloride 0.01 NaOH and/or HCI q.s. to pH 6.5 Purified Water q.s. to 100 * Aminosoap LYC-12 ,o Ingredient Concentration (%) Betaxolol HCI 0.28 Amberlite IRP-69 0.25 Carbopol 974P 0.45 Boric Acid 0.4 Mannitol 4.5 Edetate Disodium 0.01 Benzalkonium Chloride 0.01 NaOH and/or HCI q.s. to pH 6.5 Purified Water q.s. to 100 Ingredient Concentration (%) Betaxolol HCI 0.28 Benzalkonium Chloride 0.01 Carbopol 974P 0.45 Amberlite IRP-69 0.25 Triethanolamine Cocoyl Glutamate*0.03 Boric Acid 0.4 Mannitol 4.5 Edetate Disodium 0.01 NaOH and/or HCI q.s. to pH 6.5 Purified Water q.s. to 100 *Amisoft CT-12 W~ 01/21209 CA 02391976 2002-02-26 PCT/LTS00/24765 Ingredient Concentration (%) Betaxolol HCI 0.28 Benzalkonium Chloride 0.01 + 10% xs Sodium Lauroyl Glutamate* 0.03 Carbopol 974P 0.45 Amberlite IRP-69 0.25 Boric Acid 0.4 Mannitol 4.15 Edetate Disodium 0.01 Tromethamine q.s. to pH 6.5 Purified Water q.s. to 100 *Amisoft LS-11 Ingredient Concentration (%) Betaxolol HCI 0.28 Benzalkonium Chloride 0.01 + 10% xs Sodium Myristoyl Glutamate* 0.03 Carbopol 974P 0.45 Amberlite IRP-69 0.25 Boric Acid 0.4 Mannitol 4.15 Edetate Disodium 0.01 Tromethamine q.s. to 6.5 Purified Water q.s. to 100 *Amisoft MS-11 WO ~l/212~9 CA 02391976 2002-02-26 PCT/USO~/24765 Ingredient Concentration (%) Betaxolol HCI 0.28 Benzalkonium Chloride 0.01 + 10% xs Sodium Stearoyl Glutamate* 0.03 Carbopol 974P 0.45 Amberlite IRP-69 0.25 Boric Acid 0.4 Mannitol 4.15 Edetate Disodium 0.01 Tromethamine q.s. to pH 6.5 Purified Water q.s. to 100 *Amisoft HS-11 Ingredient Concentration (%) Betaxolol HCI 0.28 Benzalkonium Chloride 0.01 + 10% xs Carbopol 974P 0.45 Triethanolamine Cocoyl Glutamate*0.03 Amberlite IRP-69 0.25 Boric Acid 0.4 Mannitol 4.5 Edetate Disodium 0.01 Tromethamine q.s. to pH 6.5 Purified Water q.s. to 100 *Amisoft CT-12 Ingredient Concentration (%) Benzalkonium Chloride 0.01 Mannitol 5 Purified Water q.s. to 100 Ingredient Concentration (%) Benzalkonium Chloride 0.01 Boric Acid 0.4 Mannitol 4.9 Tromethamine 0.726 Edetate Disodium 0.01 Purified Water q.s. to 100 ,o Antimicrobial preservative effectiveness was determined using an organism challenge test according to the methods described in the United States Pharmacopeia (USP) and European Pharmacopoeia (Ph.Eur.).
Samples were inoculated with known levels of one or more of the following:
gram-positive (Staphylococcus aureus ATCC 6538) and gram-negative ,s (Pseudomonas aeruginosa ATCC 9027 and Escherichia coli ATCC 8739) vegetative bacteria, yeast (Candida albicans ATCC 10231 ) and mold (Aspergillus nigerATCC 16404). The samples were then pulled at specified intervals to determine if the antimicrobial preservative system was capable of killing or inhibiting the propagation of organisms purposely introduced into the 20 formulation. The rate or level of antimicrobial activity determined compliance with the USP and/or Ph.Eur. preservative efficacy standards for ophthalmic preparations.

The compendia) preservative standards for ophthalmic preparations are presented below:
For Bacteria:
Log Reduction of Organism Population Time Pull USP Ph.Eur. Ph.Eur.
A B
(Target) (Min) 6 hours - 2 -24 hours - 3 1 7 days - - 3 14 days 3 - -28 days NI NR NI

For Fungi:
,0 Time Pull USP Ph.Eur. Ph.Eur.
A B
(Target) (Min) 7 days - 2 -14 days N I - 1 28 days NI NI NI

NR - No organisms recovered NI - No increase at this or any following time pulls - - No requirement at this time pull The results of the microorganism challenge tests are shown in Tables 1 and 2 below.

Preservative Efficacy Standard Formulation USP Ph.Eur. B Ph.Eur. A
(Minimum) (Target) Example 1 * Pass Pass Fail Example 2 Pass Pass Fail Example 4* Pass Pass Fail Comp. Ex. 1 Pass ~ Fail Fail *Projected results based on S. aureus, P. aeruginosa and A. niger data Organism (7 day results - log reduction) Formulation S. aureus P. aeruginosaE. coliC. albicansA. nicer Comp. Ex. 2.9 5.0 -- -- 1.3 Comp. Ex. 0.0 5.0 -- -- 2.9 Comp. Ex. 1.5 5.0 -- -- 3.5 Comp. Ex. 2.1 5.0 -- -- 3.6 Comp. Ex. 0.5 5.0 5.0 1.6 2.0 Comp. Ex. 4.8 4.7 4.9 4.7 2.5 Comp. Ex. 4.8 4.7 4.9 4.7 3.7 ,o The invention in its broader aspects is not limited to the specific details shown and described above. Departures may be made from such details within the scope of the accompanying claims without departing from the principles of the invention and without sacrificing its advantages.

Claims (19)

We claim:

1. ~A method of improving or enhancing the antimicrobial efficacy of a topically administrable pharmaceutical composition comprising a cationic antimicrobial, an anionic polyelectrolyte and an active ingredient, wherein the method comprises adding to the composition an antimicrobial-enhancing amount of a fatty acid/amino acid soap having a fatty acid component derived from a C8 - C24 fatty acid and an amino acid component selected from the group consisting of lysine and arginine.

2. ~The method of Claim 1 wherein the fatty acid component is selected from the group consisting of cocoyl; linoleoyl; lauroyl; myristoyl; stearoyl;
oleoyl; and pelargodoyl fatty acid residues.

3. ~The method of Claim 2 wherein the fatty acid component is cocoyl.

4. ~The method of Claim 1 wherein the antimicrobial-enhancing amount of fatty acid/amino acid soap is from about 0.001 to about 1%.

5. ~The method of Claim 4 wherein the antimicrobial-enhancing amount of fatty acid/amino acid soap is from about 0.01 to about 0.2%.

6. ~The method of Claim 5 wherein the antimicrobial-enhancing amount of fatty acid/amino acid soap is from about 0.03 to about 0.12%.

7. ~The method of Claim 1 wherein the anionic polyelectrolyte is selected from the group consisting of: carboxyvinyl polymers; xanthan gum;
polystyrene sulfonic acid polymers; and cationic exchange resins.

8. ~The method of Claim 1 wherein the topically administrable pharmaceutical composition further comprises one or more active ingredients selected from the group consisting of ophthalmic; dermatological; otic; and nasal agents.

9. The method of Claim 8 wherein the topically administrable pharmaceutical composition comprises an ophthalmic agent selected from the group consisting of anti-glaucoma agents; anti-infectives; non-steroidal and steroidal anti-inflammatories; proteins; growth factors; and anti-allergic agents.

10. The method of Claim 8 wherein the topically administrable pharmaceutical composition further comprises one or more ingredients selected from the group consisting of pharmaceutically acceptable buffers;
tonicity agents; comfort-enhancing agents; solubilizing aids; pH adjusting agents; antioxidants; and stabilizing agents.

11. A topically administrable pharmaceutical composition comprising a cationic antimicrobial, an anionic polyelectrolyte, an active ingredient, and an antimicrobial-enhancing amount of a fatty acid/amino acid soap having a fatty acid component derived from a C8 - C24 fatty acid and an amino acid component selected from the group consisting of lysine and arginine.

12. The composition of Claim 11 wherein the fatty acid component is selected from the group consisting of cocoyl; linoleoyl; lauroyl; myristoyl;
stearoyl; oleoyl; and pelargodoyl fatty acid residues.

13. The composition of Claim 12 wherein the fatty acid component is cocoyl.

14. The composition of Claim 11 wherein the antimicrobial-enhancing amount of fatty acid/amino acid soap is from about 0.001 to about 1%.

15. The composition of Claim 14 wherein the antimicrobial-enhancing amount of fatty acid/amino acid soap is from about 0.01 to about 0.2%.

16. The composition of Claim 15 wherein the antimicrobial-enhancing amount of fatty acid/amino acid soap is from about 0.03 to about 0.12%.

17. The composition of Claim 11 wherein the anionic polyelectrolyte is selected from the group consisting of: carboxyvinyl polymers; xanthan gum;
polystyrene sulfonic acid polymers; and cationic exchange resins.

18. The composition of Claim 11 wherein the active ingredient is selected from the group consisting of anti-glaucoma agents; anti-infectives; non-steroidal and steroidal anti-inflammatories; proteins; growth factors; and anti-allergic agents.

19. The composition of Claim 11 wherein the composition further comprises one or more ingredients selected from the group consisting of pharmaceutically acceptable buffers; tonicity agents; comfort-enhancing agents; solubilizing aids; pH adjusting agents; antioxidants; and stabilizing agents.