AU2012200443B2

AU2012200443B2 - Deactivants for dust mite allergens

Info

Publication number: AU2012200443B2
Application number: AU2012200443A
Authority: AU
Inventors: Gay Cornelius; Malcolm Tom Mckechnie; Janette Suh; Ian Andrew Thompson
Original assignee: Reckitt Benckiser UK Ltd
Current assignee: Reckitt Benckiser UK Ltd
Priority date: 1997-09-25
Filing date: 2012-01-25
Publication date: 2013-10-03
Anticipated expiration: 2018-09-22
Also published as: AU2012200443A1

Abstract

A method for deactivating a Der-f and/or a Der-p allergen comprising contacting the allergen with a deactivating effective amount of a deactivant, the deactivant being hinokitiol

Description

- 1 AUSTRALIA PATENTS ACT 1990 COMPLETE SPECIFICATION FOR A STANDARD PATENT ORIGINAL Name of Applicant: Reckitt Benckiser (UK) Limited Actual Inventors: Janette Suh and Malcolm Tom McKechnie and Gay Cornelius and Ian Andrew Thompson Address for Service is: SHELSTON IP 60 Margaret Street Telephone No: (02) 9777 1111 SYDNEY NSW 2000 Facsimile No. (02) 9241 4666 CCN: 3710000352 Attorney Code: SW Invention Title: Deactivants for dust mite allergens Details of Original Application No. 2010200851 dated 05 Mar 2010 The following statement is a full description of this invention, including the best method of performing it known to me/us: File: 67249AUP02 An alternative method of creating an allergen-free environment has been to denature the allergen, for example as described in US Patent No. 4,806,526. The only effective method however of which we are aware is to apply tannic acid to the allergen. However, tannic acid can cause staining, and this is a particularly acute problem for light coloured carpets (e.g. white and light beige carpets) and other textile surfaces as tannic acid leaves a deep brown stain. Any discussion of the prior art throughout the specification should in no way be considered as an admission that such prior art is widely known or forms part of common general knowledge in the field. It is an object of the present invention to overcome or ameliorate at least one of the disadvantages of the prior art, or to provide a useful alternative. Therefore, we have been looking for allergenic denaturants which will not stain susceptible surfaces such as carpets and still deactivate the allergen. We have discovered a number of allergen deactivants which are effective against both the Der-f and the Der-p species. Quite surprisingly, we have discovered that some of these deactivants are specific to the type of dust mite allergen being treated. For example an effective Der-f allergen deactivants will not automatically work effectively as a Der-p allergen deactivant. According to a first aspect, the invention provides a method for deactivating a Der-f and/or a Der-p allergen comprising contacting the allergen with a deactivating effective amount of a deactivant, the deactivant being diazolidinyl urea. According to a second aspect, the invention provides a method for deactivating a Der-f allergen comprising contacting the allergen with a deactivating effective amount of a deactivant, the deactivant being diazolidinyl urea. According to a third aspect, the invention provides a method for deactivating a Der-p allergen comprising contacting the allergen with a deactivating effective amount of a deactivant, the deactivant being diazolidinyl urea. According to a fourth aspect, the invention provides a method for deactivating allergens deriving from Der-f and/or Der-p dust mites, said allergens being associated with faecal particles excreted by said mites on the surfaces of fabric materials selected from rugs, carpet and upholstered furniture, which method comprises applying to said fabric materials a deactivant, the deactivant being diazolidinyl urea, at an application rate of from 16 grams to 170 grams of deactivant per 10 square meters. According to a fifth aspect, the invention provides an aerosol composition when used for deactivating a Der-f and/or a Der-p allergen comprising: a) a deactivant, the deactivant being diazolidinyl urea, b) a propellant; and c) optionally, a solvent. Unless the context clearly requires otherwise, throughout the description and the claims, the words "comprise", "comprising", and the like are to be construed in an inclusive sense as opposed to an exclusive or exhaustive sense; that is to say, in the sense of "including, but not limited to". According to the invention there is provided a method for deactivating allergens derived from the Der-f and/or Der-p dust mite species, which comprises contacting the allergen with a deactivating effective amount of one or more of deactivants (herein after defined as the deactivant). The deactivants effective against one or both of Der-f allergens and Der-p allergens are: i) cedarwood oil, ii) hexadecyltrimethylammonium chloride, iii) aluminium chlorohydrate, iv) I -propoxy-propanol-2, v) polyquatemium-10 vi) silica gel, vii) propylene glycol alginate, viii) ammonium sulphate, ix) hinokitiol, 5 X) L-ascorbic acid, xi) "immobilised tannic acid", (hereinafter defined) xii) chlorohexidine, xiii) maleic anhydride, 10 xiv) hinoki oil, xv) a composite of AgCl and TiO xvi) diazolidinyl urea, xvii) 6-isopropyl-m-cresol, xviii) a compound of formula I 0 OoctyI NaO S O octyI 0 xix) the compound of formula II

OCH

3 o CH 2

OCH

3 3 xx) a polymeric dialdehyde containing two or more of -a recurring unit of the formula III

CH

2 OH CHO CHO n 5 where n 2 to 200, xxi) urea, xxii) cyclodextrin, xxiii) hydrogenated hop oil, xxiv) polyvinylpyrrolidone, 10 xxv) N-methylpyrrolidone, xxvi) the sodium salt of anthraquinone, Xxvii) potassium thioglycolate, and xxviii) glutaraldehyde Deactivants (i) through (xx) are effective against both 15 Der-f and Der-p allergens. Deactivants (xxi) through (xxvi) are effective against Der-f allergens only. Deactivants (xxvii) and (xxviii) are effective against Der-p allergens only. A compound of formula I is commercially available as 20 Aerosol OT. The compound of formula II is commercially available as parsley camphor. Hinoki oil is a mixture of thujan-3-one, 2-pinene, 3,5,7,3',4'-pentahydroflavanone and 1,3,3-trimethyl-2 25 norcamphanone. 4 Hydrogenated Hop Oil is the potassium salt of tetrahydroiso humulinic acid (also known as reduced isomerised hop extract). Propylene glycol alginate is 5 OH (HR)OOC OH (HR)OOC OH HO ~ OO OH COO(RH) n Chlorohexadine is 1,1' -hexamethylenebis (5- (4 chlorophenyl)]-biguanide. Hinokitol is 0-thujaplicin, a compound of the formula 10 0 / OH

CH

3

CH

3 Germall II is diazolidinylurea. Thymol is 6 -isopropyl-m-cresol. Cedarwood oil contains a- and P- cedrene (ca 80%), cedrol (3-14%) and cedrenol. Other sesquiterpenes and 15 some monoterpenes are also present. 5 Polyquaternium-10 is. a polymeric quaternary ammonium salt of hydroxyethyl cellulose reacted with a trimethyl ammonium substituted epoxide commercially available as Polymer JR-125. 5 Silica gel is also known as colloidal silica or silicic acid and is commercially available as Kent. "Immobilised tannic acid" is tannic acid on polyvinyl pyrrolidone beads. Immobilised Tannic Acid was prepared as follows: 100 mg of tannic acid was dissolved 10 in water; So mg of Polyclar 10 (ISP, Guildford Surrey) polyvinyl pyrrolidone beads were added and stirred for one hour; the beads were filtered off the solution and washed with a few mis of iced water until no colour was seen in the washings; they were then dried in the oven at 15 50 0 C. The composite of silver chloride and TiO 2 is made up of 20% wt/wt AgCl on 80% TiO 2 3-5 pm porous beads. In compositions containing the deactivant, the deactivant is present in an amount of from 0.01% to 7%, 20 preferably from 0.01% to 3%. In methods for treating rugs and carpets to deactivate allergents, the amount of deactivant present is from about 16gm to about 170gm per 10 square meters, preferably about 32gm per 10 square meters. 25 Preferably the deactivant is selected from xiv) hinoki oil, xv) a composite of AgCl and

T.O

2 , xvi) diazolidinyl urea xvii) 6-isopropyl-m-cresol, 30 xii) chlorohexidine, Xiii) maleic anhydride, 6 xxvi) the sodium salt of anthraquinone and xviii) a compound of formula I or II, defined above, and xix) a compound of formula II, defined above. 5 Further according to the invention there is provided an aerosol composition containing i) cedarwood oil, ii) hexadecyltrimethylammonium chloride, iii) aluminium chlorohydrate, 10 iv) 1-propoxy-proparol-2, v) polgquaternium--.o vi) silica gel, vii) propylene glycol alginate, viii) ammonium sulphate, is ix) hinokitiol, X) L-ascorbic acid, xi) "immobilised tannic acid", (hereinafter defined) xii) chlorohexidine, 20 xiii) maleic anhydride, xiv) hinoki oil, xv) a composite of AgCl and :iO. xvi) diazolidinyl urea, xvii) 6 -isopropyl-m-crescl, 25 xviii) a compound of fcrmIa I 0 .1octi Nal %S O octyi 0 7 xix) the compound of formula II

OCH

3 O

CH

2

OCH

3 xx) a polymeric dialdehyde containing two or more of a recurring unit of the 5 formula III

CH

2 OH 0 O O CHO CHO n where n 2 to 200, xxi) urea, xxii) cyclodextrin, 10 xxiii) hydrogenated hop oil, xxiv) polyvinylpyrrolidone, xxv) N-methylpyrrolidone, xxvi) the sodium salt of anthraquinone, xxvii) potassium thioglycolate, and 15 xxviii) glutaraldehyde b) a propellant, and c) optionally, a solvent. 8 Preferably the amount of deactivant present in such a composition is from 0.01% to 7%, more preferably 0.01% to 3%, Preferably the amount of propellant present in such 5 a composition is from 4% to 50%, more preferably from 4% to 30%, Preferably the amount of solvent present in such a composition is 0% to 99.95%, more preferably 0% to 90%, and most preferably from 20% to 9C%. 10 Preferably the deactivant in such aerosol composition is selected from hinoki oil, a composite of AgCl with TiO., diazolidinyl urea, 15 6-isopropyl-m-cresol, chlorohexidine, maleic anhydride, the sodium salt of anthracuin~ne, and a compound of formula I or I: defined above. 20 Preferably the propellant is selected from those commercially available, for example C .. 4 alkanes, chlorofluorocarbons and compressed gases such as nitrogen, air and carbon dioxide. Preferably the solvent is selected from C 1

-

6 alcohols 25 (e.g. ethanol) or water. In addition, the compositions of this invention may also contain one or more of the fcllcwing: a fragrance, preferably in an amount of 0% to 5%, more preferably 0% to 2%; 9 an antimicrobial compound e.g. alkyldimethylbenzyl ammonium saccharinate, preferably in an amount of 0.01% to 1%; a surfactant, e.g. Dow Corning 193 Surfactant, 5 preferably in an amount of 0.01% to 1%; a corrosion inhibitor, e.g. sodium nitrite, sodium benzoate, triethanolamine and ammonium hydroxide, preferably in an amount of 0.01% to 10%; and 10 a miticide, such as benzyl benzoate, pyrethroid pemethrin, d-allethrin and optionally a synergist such as piperonyl butoxide, preferably in an amount of 0.1% to 10%. It has been found that deactivants of the invention 15 have as effective allergen deactivating properties as tannic acid but without the drawback of staining. The invention will now be illustrated by the following Examples. Examples 20 The test procedure in Examples 1 to 55 is as follows and is known as the ELISA protocol. The ELISA protocol for Der-f and Der-p has been developed as follows as a measure of denaturing property for denaturants. 25 ELISA Protocol 1 1. Dust is collected from Hoover' vacuum cleaner bags and passed through a series of sieves down to 63 microns. 10 2. Clean petri dishes are labelled with the chemical to be tested (on the base). Three replicates are used for each treatment. 3. Filter paper is used to line each dish and 0.2g of 5 Aus s added to each dish onto the filter paper. The lid (or base, as dishes are inverted) is replaced and the dish is shaken to disperse the dust evenly over the filter paper. 4. 2% aqueous solutions of deactivant were used except 10 for the silver chloride composite where 0.05% was used instead. Immobilised tannic acid was used as a 1% dispersion. The hydrogenerated hop end was used at the 2% level (in the form of a 10% solution). Water insoluble deactivants were emulsified with a sorbitone 15 oleate surfactant (i.e. Tween) . Hinokitol was used at 0.5% not 2%. 5. The dust is sprayed with the corresponding treatment, 2 sprays are required for sufficient coverage(l spray = 1.5 ml). 20 6. Leave uncovered at room temperature, in well aerated room, until filter paper is dry. This can cake up to 4 hours. 7. Empty dust in epindorfs labelled according to treatment. 25 8. Add 1 ml of 5% Bovine Serum Albumen Phosphate Butter Saline - Tween BSA-PBS-T to each epindorf (5 times the weight of dust) (20ml of BSA-PBS-T =1 g of BSA in 20ml of PBS-T). 9. Leave overnight in a refrigerator. 30 10. Centrifuge for 5 minutes at 13,000 rpm. 11 11. Decant the supernatant into a new epindorf labelled according to treatment. 12. Centrifuge again for 5 minutes at 13,000 rpm. 13. Make up dilutions of 1:10 and 1:100 by adding 100 Ai 5 of neat solution to 900 pl of 1% BSA-PBS-T (1:10) . This is repeated using 100 y1 of 1:10 dilution and add to 900 gl of 1% BSA-PBS-T for 1:100 dilution. ELISA Protocol 2 for Der-f and Der-p: Indoor Biotechnologies 10 1. Prepare samples and dilutions as in protocol 2. Prepare 500 ml of 50 mM carbonate/bicarbonate buffer by dissolving 0.

7 959 Na 2 CO and 1.4659 NaHCO, in 500 ml of distilled water. Check the pH is at 9.6. (This solution is kept in the refrigerator in a conical flask) 15 3. Monoclonal antibody (kept in the freezer) has to be added to the buffer using the following method, (l mg per well; 11 ml is needed) applied to the ELISA plate - llml of carbonate/bicarbonate buffer is added to the dispensing tray. 20

-

11p1 of Der-fl or Der-pl monoclonal antibody (Stored in freezer, epindorf in use is in the refrigerator) is added to the buffer. To ensure that all the antibody is removed from the tip, wash out the pipette tip by sucking up and down I the buffer solution, 25 gently stirring to mix thoroughly. 4. Pipette 100 l of the antibody solution into each well of the microtiter plate, cover with a plate sealer and leave overnight at 4 0 C. 12 5. Empty the plate by quickly inverting it over the sink, then dry by banging on a stack of paper towels. 6. Add 200 l of wash buffer to each well: PBS/0/05% tween (PBS-T). 5 7. Repeat stages 5 and 6 once more (making a total of 2 washes). 8. Make sure all the wells are dry, then add 100 pl of 1% BSA-PBS-T. Replace the plate sealer and incubate for 1 hour at room temperature*. 10 9. Repeat steps 5 to 7 (2 washes) 10. *During the hour incubation period, prepare the allergen standards at dilutions between 125 and 1 pg/ml Der f 1 or Der pl: - Add 25 pl of allergen standard (kept in the 15 refrigerator in polystyrene box) to 475 pl of 1% PBS-BSA-T and mix thoroughly - labelled '125'. - 250 pl of 1% PBS-BSA-T is added to 7 further epindorfs which are labelled 62.5, 31.25, 15.63, 7.61, 3.9, 1.95 and 0.98. 20 - 250 p1 is taken from the 1st epindorf (labelled 125) and transferred to the next (labelled 62.5). This is mixed thoroughly. - Using a new pipette tip, 250 p.1 is removed from epindorf labelled 62.5 and transferred to 31.25, 25 this procedure is continued down to the 0.98 concentration (125, 62.5, 31.25, 15.63, 7.61, 3.9, 1.95, 0.98)

-

In total 475 + (250 x 7) = 2.3ml : 0.023g of BSA added to 2.3 ml of PBS-T. 13 11. Add 1004l aliquots of the allergen sample to the plate along with the standard allergen samples for the reference curve in duplicate. The standards usually go in the first two columns on the left hand side, with the S least concentrated on top. Incubate for 1 hour. 12. Follow stages 5 to 6, completing a total of 5 washes. 13. Pour 11 ml of 1% BSA-PBS-T(0.llg of BSA to llml of PBS-T) to the dispensing tray. Add 11 i of the 10 biotinylated monoclonal antibody (refrigerator) and mix thoroughly. 14. Pipette 100 1 into each well and incubate for 1 hour at room temperature. 15. Empty plate and wash as described in stage 12. (5 15 washes). 16. Add 11 #1 of Streptavidin (freezer) to 11 ml of 1%BSA-PBS-T. Pipette 100 pl into each well and incubate for 30 minutes. Reserve any remaining solution in a vial. 20 17. Empty plate and wash as described in stage 12 (5 washes). 18. Make a solution of OPD, by putting the two tablets (in silver and gold foil) into 20 ml of distilled water (in a glass vial) . Shake quite vigorously in the dark 25 until the tablets have dissolved (Wrap the vial up either in tin foil or paper towel). 19. Add a small amount to the remaining solution from stage 16. Wait for a colour change (positive reaction). Add 200 .1 to each well and incubate for a minimum of 30 30 minutes in the dark. 14 20. Read the plate at 4 50nm/405nm if filter not available. ExamDes 1 to 26 The deactivants. as set out in the following table, 5 were used against Der-f allergens according to the above procedure and the results are as given below. Tannic acid was used as a comparator. What was measured after treatment with deactivant and tannic acid was the amount of allergen remaining active after treatment. The ratio 10 of amount of remaining active allergen after treatment with deactivant and tannic acid is also given. 15 4.1 X x x x 5 , M C0000 ' cu CD a aC 5 ot C D0k ej %^ O 00 f-00 <= t 1- -0I , M U EV too - - - - - - - EE 00 0 2 -u 'o~ -u c- Cq m X 'T % -w0 16 < ) X > o m ccw cl v 94 00 0 cl ~ ~ 4w>NF 0 o 0 0 E E 2jE 4 C 0 17 The deactivants, as set out in the following table, were used against Der-p allergens according to the above procedure and the results are as given below. What was 5 measured were the amount of allergens remaining after treatment with deactivant and the amount of allergens remaining after vacuuming with no deactivant treatment. 18 M > > : >1 "I> x .- .: 0 ONC SC Ne Va.

4) 4 = U= cc 0 @h C o 0 0 0 o O eo ': 0 .2 4.)19 W~4. .tU.ve v m m C m V E C3 oN m~ " - 0% 00I m CDU q~ "0e lt- o E cu 0 Further samples were tested as above and compared against tannic acid. The ratio of actives remaining after deactivant treatment and actives remaining after tannic acid treatment are given below: Example Deactivant atio of actives Number remaining after deactivant treatment over those remaining after tannic acid treatment 48 Germall II 1.5 vi 49 N-methyl pyrrolidone 4.0 xv 50 Hinoki Oil 4.0 iv 51 Silver chloride/Tio, 3.5 v 52 Thymol 4.0 vii 53 Chlorohexidine 3.0 T1 54 Maleic 1.0 iii anhydride 55 Glutaraldehyde 1.5 xviTT Examples 56-59 The following formulations can be made up as carrier compositions for use in an aerosol for deactivating Der-f and Der-p allergens. 21 Anhydrous Ethanol (SD Solvent Alcohol 40) 79.646 A y dimet yl benzyl Cationic Surfactant ammonium saccharinate 0.106 Corrosion Inhibitor (I) 0.192 Corrosion Inhibitor (11) 0.192 Corrosion Inhibitor (III) 0.096 Deionized Water Water/Solvent 15.768 CarbonWL Dioxide Propellant 1.6 4.000 100.000 22 Raw Inredient tion Description by Weight Anhydrous Ethanol (SD Solvent * 57.000 Alcohol 40) Fragrance#17 Fragrance 0.0500 Dow Corning 193 Surfactant 0.025 Surfactant Corrosion Inhibitor (I) 0.100 Corrosion Inhibitor (II) 0100 Deionized Water Water/solvent * 14.725 NP-40/Butane 40 Hydrocarbon 28.000 propellant ITOTAL 100.000 * May replace with 95% Ethanol (SD Alcohol 40) at 61.755% by weight and 9.970% by weight Deionized water ) 23 Raw Ingredient ic Description by Weight Any drous Et anol SD Solvent Alcohol 40) 79.646 Benzyl Benzoate - an Active/ester acaricide 4.600 AikI d imetyl b enzyl Cationic Surfactant ammonium saccharinate 0.106 Corros ion Inhibitor (I)- 0 .192 Corrosion Inhibit-or-(11)7 0.192 Crrosion Inhibitor (I-II) -- O6 Deionized Waer Water/solvent Carbon Dioxide Propellant 1116 14.000 24 Description by weight An ydrous Ethano (SD Solvent *57 .Qn Alcohol 40) Benzyl Benzoate Active/ester 4.600 Fragrance#17 - Fragrance 0.0500 Dow Corning 193 Surfactant 0.025 Surfactant Corrosion Inhibitor I) 0.100 Corrosion Inhi itor II 0.100 Deionize Water Water/solvent *10.125 NP-40/Butane 40 Hydrocarbon 28.000 propellant TOTAL_1_ 0.000 * = May replace 95% Ethanol (SD Alcohol 40) at 61.735% by weight and 5.370% by weight Deionized water. 25

Claims

1. A method for deactivating a Der-f and/or a Der-p allergen comprising contacting the allergen with a deactivating effective amount of a deactivant, the deactivant being diazolidinyl urea. 5

2. A method for deactivating a Der-f allergen comprising contacting the allergen with a deactivating effective amount of a deactivant, the deactivant being diazolidinyl urea.

3. A method for deactivating a Der-p allergen comprising contacting the allergen with a deactivating effective amount of a deactivant, the deactivant being diazolidinyl 10 urea.

4. A method for deactivating allergens deriving from Der-f and/or Der-p dust mites, said allergens being associated with faecal particles excreted by said mites on the surfaces of fabric materials selected from rugs, carpet and upholstered furniture, which method comprises applying to said fabric materials a deactivant, the deactivant being 15 diazolidinyl urea, at an application rate of from 16 grams to 170 grams of deactivant per 10 square meters.

5. A method according to claim 4 in which the allergens derive from Der-f dust mites.

6. A method according to claim 4 in which the allergens derive from Der-p dust 20 mites.

7. An aerosol composition when used for deactivating a Der-f and/or a Der-p allergen comprising: a) a deactivant, the deactivant being diazolidinyl urea, b) a propellant; and 25 c) optionally, a solvent.

8. A composition according to claim 7 in which the amount of deactivant present is from 0.01% to 7%, the amount of propellant present is from 0.05% to 3%, and the amount of solvent present is from 0% to 99.95%, all percentages being by weight.

9. A composition according to claim 7 or claim 8 in which the propellant is 5 selected from C, 4 alkane and carbon dioxide.

10. A composition according to any one of claims 7 to 9 in which the solvent is selected from C 1 - alcohols or water.

11. A composition according to claim 10 in which the solvent is ethanol.

12. A composition according to any one of claims 7 to 1I in which the composition 10 may also contain one or more of the following: a fragrance, a surfactant, an antimicrobial agent, a corrosion inhibitor, and/or 15 a miticide.

13. A method for deactivating a Der-f and/or a Der-p allergen according to any one of Claims 1 to 3 substantially as herein described with reference to any one of the embodiments of the invention illustrated in the accompanying drawings and/or examples. 20

14. A method for deactivating allergens deriving from Der-f and/or Der-p dust mites according to any one of Claims 3 to 6 substantially as herein described with reference to any one of the embodiments of the invention illustrated in the accompanying drawings and/or examples.

15. An aerosol composition when used for deactivating a Der-f and/or a Der-p 25 allergen according to any one of Claims 7 to 12 substantially as herein described with reference to any one of the embodiments of the invention illustrated in the accompanying drawings and/or examples. ')7