GB2329586A

GB2329586A - House dust mite allergen deactivation

Info

Publication number: GB2329586A
Application number: GB9720275A
Authority: GB
Inventors: Malcolm Tom Mckechnie; Gay Joyce Cornelius; Ian Andrew Thompson; Janette Suh
Original assignee: Reckitt and Colman Products Ltd
Current assignee: Reckitt Benckiser Healthcare UK Ltd
Priority date: 1997-09-25
Filing date: 1997-09-25
Publication date: 1999-03-31
Also published as: GB9720275D0; ZA988700B

Abstract

A Der-f allergen is deactivated by contacting it with a deactivating effective amount of one or more of i) urea, ii) cedarwood oil, iii) cyclodextrin, iv) hexadecyltrimethylammonium chloride v) aluminium chlorohydrate, vi) 1-propoxy-propanol-2, vii) polyquaternium-10 viii) silica gel, ix) hydrogenated hop oil, x) propylene glycol alginate, xi) polyvinyl pyrrolidone, xii) ammonium sulphate, xiii) hinokitiol, xiv) N-methyl pyrrolidone, xv) L-ascorbic acid, xvi) "immobilised tannic acid", xvii) chlorohexidine, xviii) maleic anhydride, xix) the sodium salt of anthraquinone, xx) hinoki oil, xxi) a composite of silver chloride and TiO2 xxii) diazolidinyl urea, xxiii) 6-isopropyl-m-cresol, xxiv - xvi) a compound of formula I or II herein or a polymeric compound containing two or more of a recurring unit of the formula III herein. The deactivant may be combined with a propellant and optionally a solvent to give an aerosol composition.

Description

Improvements in or relating to organic compositions It has been known for a long time that house dust can trigger allergenic reactions in humans, such as asthma and rhinitis. It was reported, as early as 1928, that it was the dust mites in the dust that were the primary source of the allergenic response but it was only in the 60's that researchers appreciated its significance.

It is believed that the faeces of the house dust mite, Dermatophagoides farinae (known as Der-f) and Dermatophagoides pteronyssinus (known as Der-p) trigger the immune responses of the body, thereby giving rise to well known allergenic symptoms.

A review of this is given in Experimental and Applied Acarology, 10 (1991) p. 167-186 in an article entitled "House dust-mite allergen" : A review by L. G.

Arlian.

One way to overcome these allergenic response has been to thoroughly vacuum surfaces, such as carpets, that contain the dust mites and their faeces thoroughly and often, but that is both time consuming (i.e. has to be regularly done if one wants to make an allergenic free environment) and is very dependant on the efficiency of vacuum cleaner and filter bag used e.g. micron filter bag or 2 layer vacuum bags.

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.

Therefore, we have been looking for allergenic denaturants which will not stain susceptible surfaces such as carpets and still deactivate the allergen.

We have surprisingly found that 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.

We have looked into Der-f allergen deactivant and have found only a select number of deactivants destroy the Der-f allergen, whilst at the same time not leaving a stain.

According to the invention there is provided a method for deactivating a Der-f allergen comprising contacting the allergen with a deactivating effective amount of one or more of deactivants (herein after defined as the deactivant) selected from i) urea, ii) cedarwood oil, iii) cyclodextrin, iv) hexadecyltrimethylammonium chloride v) aluminium chlorohydrate, vi) l-propoxy-propanol-2, vii) polyquaternium-l0 viii) silica gel ix) hydrogenated hop oil, x) propylene glycol alginate, xi) polyvinyl pyrrolidone, xii) ammonium sulphate, xiii) hinokitiol, xiv) N-methyl pyrrolidone, xv) L-ascorbic acid, xvi) "immobilised tannic acid", (hereinafter defined) xvii) chlorohexidine, xviii)maleic anhydride, xix) the sodium salt of anthraquinone, xx) hinoki oil, xxi) a composite of AgCl and TiO2 xxii) diazolidinyl urea7 xxiii)6-isopropyl-m-cresol, xxiv) a compound of formula I,

xxv) a compound of formula II

xxvi) a polymeric compound containing two or more of a recurring unit of the formula III

where n = 2 to 200.

In this Specification, the definition of the following compounds or compositions is given below: A compound of formula I is commercially available as Aerosol OT.

A 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-norcamphanone.

Cedarwood oil, contains a- and ss- cedrene (ca 80%), cedrol (3-14%) and cedrenol. Other sesquiterpenes and some monoterpenes are also present.

Polyquaternium-lO is a polymeric quaternary ammonium salt of hydroxy ethyl cellulose reacted with a trimethyl ammonium substituted epoxide commercially available as Polymer JR-125.

Silica gel is also known as colloidal silica or silicic acid and is commercially available as Kent.

Hinokitiol is also known as ss-thujaplicin.

"Immobilised tannic acid" is tannic acid on polyvinyl pyrrolidone beads. "Immobilised Tannic Acid" is prepared as follows: 100 mg of tannic acid dissolved in water, 50 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 mls of iced water until no colour was seen in the washings. They were then dried in the oven at 50"c.

The composite of silver chloride and TiO2 is made up of 20% wt/wt Agol on 80% TiO2 3-5 um porous beads.

The deactivant is present in an amount of from 0.01 - 7%, preferably 0.01 to 3%.

Preferably the amount of deactivant present in sucE a method is from 0.5 oz to 5 oz per 10 yds2 , more preferably 1 oz per 11 yds2 of area treated.

Preferably the deactivant is selected from hinoki oil, a composite of AgCl and TiO2, diazolidinyl urea 6-isopropyl-m-cresol, chlorohexidine, maleic anhydride, the sodium salt of anthraquinone and a compound of formula I or II, defined above.

Further according to the invention there is provided an aerosol composition containing a) a deactivant (hereinafter the Deactivant) selected from i) urea, ii) cedarwood oil, iii) cyclodextrin, iv) hexadecyltrimethylammonium chloride v) aluminium chlorohydrate, vi) l-propoxy-propanol-2, vii) polyquaternium-lO, viii) silica gel ix) hydrogenated hop oil, x) propylene glycol alginate, xi) polyvinyl pyrrolidone, xii) ammonium sulphate, xiii) hinokitiol, xiv) N-methyl pyrrolidone, xv) L-ascorbic acid, xvi) "immobilised tannic acid", (hereinafter defined) xvii) chlorohexidine, xviii) maleic anhydride, xix) the sodium salt of anthraquinone, xx) hinoki oil, xxi) a composite of silver chloride and TiO2 xxii) diazolidinyl urea, xxiii))6-isopropyl-m-cresol, xxiv-xxvi)a compound of formula I, II or III defined above; b) a propellant and c) optionally a solvent.

Preferably the amount of deactivant present in such a composition is from 0.01 - 7%, preferably 0.01 to 3%, Preferably the amount of propellant present in such a composition is 4-50%, more preferably 4 to 30%, Preferably the amount of solvent present in such a composition is 0 to 99.95, more preferably 0 to 90%, most preferably 20 to 90%.

Preferably the deactivant is selected from hinoki oil, a composite of AgCl with TiO2, diazolidinyl urea, 6-isopropyl-m-cresol, chlorohexidine, maleic anhydride, the sodium salt of anthraquinone and a compound of formula I or II defined above.

Preferably the propellant is selected from those commercially available, for example Cl4alkanes and hydrochlorofluorocabrons and compressed gases such as nitrogen air and carbon dioxide.

Preferably the solvent is selected from C16 alcohols (e.g. ethanol) or water.

In addition the composition may also contain one or more of the following a fragrance, (preferably in an amount of 0 to 5%), more preferably 0 to 2%. an antimicrobial compound e.g. alkyl dimethyl benzyl ammonium saccharinate (preferably in an amount of 0.01 to 1%) a surfactant (e.g. Dow Corning 193 Surfactant or (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/or a miticide (such as benzyl benzoate, pyrethroid pemethrin, d-allethrin and optionally a synergist such as pipernoyl butoxide (preferably in an amount of 0.1 to 10%)).

It has been found that deactivants of the invention 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.

The test procedure in Examples 1 to 17 is as follows and is known as the ELISA protocol.

The ELISA protocol for Der-f has been developed as follows as a measure of denaturing property for denaturants.

ELISA Protocol 1 1. Dust is collected from Hoover (a trademark) bags and passed through a series of sieves down to 63 microns.

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 dust is 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 of dust evenly over the filter paper.

4. 2W aqueous solutions of deactivant was used except 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 deactivant were emulsified with surfactant (a sorbitone 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(1 spray = 1.5ml).

6. Leave uncovered at room temperature, in well aerated room, until filter paper is dry. This can take up to 4 hours.

7. Empty dust in epindorfs labelled according to treatment.

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 =lg of BSA in 20ml of PBS-T).

9. Leave overnight in the fridge.

10. Centrifuge for 5 minutes at 13,000 rpm.

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 dilution's of 1:10 and 1:100 by adding 100ul of neat solution to 900ul of 1% BSA-PBS-T (1:10).

This is repeated using 100ul of 1:10 dilution and add to 900ul of 1% BSA-PBS-T for 1:100 dilution.

ELISA Protocol 2 for Der f 1: Indoor Biotechnologies 1. Prepare samples and dilutions as in protocol 1.

2. Prepare 500 ml of 50 mM carbonate/bicarbonate buffer by dissolving 0.795g Na2CO3 and 1.465g NaHCO3 in 500ml of distilled water. Check the pH is at 9.6.

(This solution is kept in the fridge in a conical flask) 3. Monoclonal antibody, this is kept in the freezer.

(lug per well ; llml is needed) has to be added to the buffer using the following method this is applied to the ELISA plate: - llml of carbonate/bicarbonate buffer is added to the dispensing tray.

- lip1 of Der f 1 monoclonal antibody (Stored in freezer, epindorf in use is in the fridge) 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, gently stirring to mix thoroughly.

4. Pipette 100p1 of the antibody solution into each well of the microtiter plate, cover with a plate sealer and leave overnight at 49C.

5. Empty the plate by quickly inverting it over the sink, then dry by banging on a stack of paper towels.

6. Add 200p1 of wash buffer to each well: PBS/0/05% tween (PBS-T).

7. Repeat stages 5 and 6 once more (making a total of 2 washes).

8. Make sure all the wells are dry, then add 100pl of 1% BSA-PBS-T. Replace the plate sealer and incubate for 1 hour at room temperature*.

9. Repeat steps 5 to 7 (2 washes).

10. +During the hour incubation period, prepare the allergen standards at dilutions between 125 and 1 ng/ml Der f 1: - Add 25pl of allergen standard (kept in the fridge in polystyrene box) to 475p1 of 1% PBS-BSA-T and mix thoroughly - labelled '125'.

- 250pl of 1% PBS-BSA-T is added 7 further epindorfs which are labelled 62.5, 31.25, 15.63, 7.61, 3.9, 1.95 and 0.98.

- 250pl 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, 250p1 is removed from epindorf labelled 62.5 and transferred to 31.25, 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+(250x7)=2.3ml : 0.023g of BSA added to 2.3ml of PBS-T.

11. Add 100p1 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 least concentrated on top.

Incubate for 1 hour.

12. Follow stages 5 to 6, completing a total of 5 washes.

13. Pore liral of 1% BSA-PBS-T(O.llg of BSA to llml of PBS-T) to the dispensing tray. Add lip1 of the biotinylated monoclonal antibody (fridge) and mix thoroughly.

14. Pipette lOOp1 into each well and incubate for 1 hour at room temperature.

15. Empty plate and wash as described in stage 12. (5 washes).

16. Add ilul of Streptavidin (freezer) to llml of 1%BSA-PBS-T. Pipette lOOp1 into each well and incubate for 30 minutes. Reserve any remaining solution in a vial.

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 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 200pl to each well and incubate for a minimum of 30 minutes in the dark.

20. Read the plate at 450nm/405nm if filter not available.

Examples 1 to 26 The deactivants, as set out in the following table, were treated 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 deactivation tannic acid was the amount of allergen remaining active after treatment. The ratio of amount of remaining active allergen after treatment with deactivant tannic acid is also given.

Table

Example Deactivant Amount of Allergen Amount of Ratio of remaining Number remaining active after Allergen active allergen deactivant treatment remaining active after after tannic acid Deactivant/Tannic treatment Acid Treatment 1 Urea 3750 1500 2.500 i 2 Polymeric dialdehyde 1325 550 2.409 xxvi 3 Cedarwood oil 1800 750 2.400 ii 4 Cyclodextrin 3850 1700 2.265 iii 5 hexadecyltrimethylammonium chloride 4075 1800 2.264 iv 6 Aluminium chlorohydrate 1675 750 2.233 v 7 1-propoxy-propanol-2 3950 1800 2.194 vi 8 Silica Gel (Kent) 2037.5 933.5 2.183 viii 9 polyquaternium-10 (Polymer JR-125) 4335 2000.00 2.168 vii 10 Hydrogenated Hop Oil 1100 550 2.000 ix 11 Propylene glycol alginate 3175 1700 1.868 x 12 Poly vinyl pyrrolidone 2450 1425 1.719 xi 13 Ammonium sulphate 2750 1700 1.618 xii

Example Deactivant Amount of Allergen Amount of Ratio of remaining Number remaining active after Allergen active allergen deactivant treatment remaining active after after tannic acid Deactivant/Tannic treatment Acid Treatment 14 Hinokitol (0.5%) 3065 2000.00 1.533 xiii 15 N-methyl pyrrolidone 1600 1175 1.362 xiv 16 L-Ascorbic Acid 2000 1500 1.333 xv 17 Immobilised Tannic Acid 1550 1175 1.319 xvi 18 Aerosol OT 1525 1175 1.298 xxiv 19 Chlorohexidine 1412.5 1425 0.991 xvii 20 Parsley Camphor 1225 1387.5 0.883 xv 21 Maleic anhydride 1312.5 1500 0.875 xxviii 22 Anthraquinone sodium salt 1530 2000 0.765 xix 23 Hinoki oil 1025 1387.5 0.739 xx 24 Composite of AgCl and TiO2 1025 1425 0.719 xxi 25 Germall II 950 1387.5 0.685 xxii 26 Thymol 725 1387.5 0.523 xxiii In the table certain compounds are used that are defined as follows: Hydrogenated Hop Oil is the potassium salt of tetrahydroiso humulinic acid (known as reduced isomerised hop extract).

Polymeric dialdehyde is a compound containing 2-200 recurring units of the formula III.

Propylene glycol alginate is

Chlorohexadene is l,1'-hexamethylene bis [5-(4-chlorophenyl)-biguanide Hinokitiol is -thujaplin, a compound of the formula

Aerosol OT is a compound of the formula

Parsley extract is a compound of the formula

Germall II is diazolidinyl urea and Thymol is 6-isopropyl -m- cresol Examples 27 to 30 The following formulations can be made up as a compositions for use as an aerosol for deactivating der-f allergens). EXAMPLE 27

Raw Ingredient Description Item Classification % By Weight Anhydrous Ethanol (SD Solvent Alcohol 40) 79.646 Alkyl dimethyl benzyl Cationic Surfactant ammonium saccharinate 0.106 Corrosion Inhibitor 0.192 Corrosion Inhibitor 0.192 Corrosion Inhibitor 0.096 Deionized Water Water/Solvent 15.768 Carbon Dioxide Propellant 4.000 TOTAL 100.000 EXAMPLE 28

Raw Ingredient Item Classification % Description by Weight Anhydrous Ethanol (SD Solvent * 57.000 Alcohol 40) Fragrance&num;17 Fragrance 0.0500 Dow Corning 193 Surfactant 0.025 Surfactant Corrosion Inhibitor 0.100 Corrosion Inhibitor 0.100 Deionized Water Water/solvent * 14.725 NP-40/Butane 40 Hydrocarbon 28.000 propellant TOTAL 100.000 * = May replace with 95% Ethanol (SD Alcohol 40) at 61.755% by weight and 9.970% by weight Deionized water EXAMPLE 29

Raw Ingredient Item Classification Description by Weight Anhydrous Ethanol (SD Solvent 79.646 Alcohol 40) Benzyl Benzoate - an Active/ester 4.600 acaricide Alkyl dimethyl benzyl Cationic Surfactant 0.106 ammonium saccharinate Corrosion Inhibitor 0.192 Corrosion Inhibitor 0.192 Corrosion Inhibitor 0.096 Deionized Water Water/solvent 11.168 Carbon Dioxide Propellant 4.000 TOTAL 100.000 EXAMPLE 30

Raw Ingredient Item Classification % Description by weight Anhydrous Ethanol (SD Solvent *57.000 Alcohol 40) Benzyl Benzoate Active/ester 4.600 Fragrance&num;17 Fragrance 0.0500 Dow Corning 193 Surfactant 0.025 Surfactant Corrosion Inhibitor 0.100 Corrosion Inhibitor 0.100 Deionized Water Water/solvent *10.125 NP-40/Butane 40 Hydrocarbon 28.000 propellant TOTAL 100.000 * = May replace 95% Ethanol (SD Alcohol 40) at 61.755% by weight and 5.370% by weight Deionized water.

Claims

CLAIMS 1. A method for deactivating a Der-f allergen comprising contacting the allergen with a deactivating effective amount of one or more of deactivants (herein after defined as the Deactivant) selected from i) urea,

ii) cedarwood oil,

iii) cyclodextrin,

iv) hexadecyltrimethylammonium chloride

v) aluminium chlorohydrate,

vi) 1-propoxy-propanol-2,

vii) polyquaternium-l0

viii) silica gel

ix) hydrogenated hop oil,

x) propylene glycol alginate,

xi) polyvinyl pyrrolidone,

xii) ammonium sulphate,

xiii) hinokitiol,

xiv) N-methyl pyrrolidone,

xv) L-ascorbic acid,

xvi) "immobilised tannic acid", (hereinafter defined)

xvii) chlorohexidine,

xviii) maleic anhydride, xix) the sodium salt of anthraquinone,

xx) hinoki oil,

xx) a composite of silver chloride and TiO2

xxi) diazolidinyl urea,

xxii) 6-isopropyl-m-cresol,

xxiv)a compound of formula I,

xxv) a compound of formula II

and

xxvi) a polymeric compound containing two or more of a recurring unit of the formula III

where n = 2 to 200 2. A method according to Claim 1 in which the amount of Deactivant present is from 0.5 oz to 5 oz per 10 yds? 3. A method according to Claim 1 or Claim 2 in which the Deactivant is selected from hinoki oil, a composite of AgCl with TiO2, diazolidinyl urea 6-isopropyl-m-cresol, chlorohexidine, maleic anhydride, the sodium salt of anthraquinone and a compound of formula I or II, defined in Claim 1.

4. An aerosol composition containing a) a deactivant selected from i) urea,

ii) cedarwood oil,

iii) cyclodextrin,

iv) hexadecyltrimethylammonium chloride

v) aluminium chlorohydrate,

vi) 1-propoxy-propanol-2,

vii) polyquaternium-l0

viii) silica gel

ix) hydrogenated hop oil,

x) propylene glycol alginate,

xi) polyvinyl pyrrolidone,

xii) ammonium sulphate,

xiii) hinokitiol,

xiv) N-methyl pyrrolidone,

xv) L-ascorbic acid, xvi) "immobilised tannic acid", (hereinafter defined) xvii) chlorohexidine, xviii)maleic anhydride, xix) the sodium salt of anthraquinone, xx) hinoki oil, xxi) a composite of silver chloride and TiO2 xxii) diazolidinyl urea, xxiii)6-isopropyl-m-cresol, xxiv a compound of formula I,

xxv) a compound of formula II

and

xxiv) a polymeric compound containing two or more of a recurring unit of the formula III

where n = 2 to 200 (hereinafter defined as the Deactivant). b) a propellant and c) optionally a solvent.

6. A composition according to Claim 5 in which the amount of Deactivant present in such a composition is from 0.01 to 7%, the amount of propellant present in such a composition is 0.05 to 3%, and the amount of solvent present in such a composition is O to 99.95%.

7. A composition according to Claim 5 or Claim 6 in which the Deactivant is selected from hinoki oil, a composite of AgCl with TiO2, diazolidinyl urea, 6-isopropyl-m-cresol, chlorohexidine, maleic anhydride, the sodium salt of anthraquinone and a compound of formula I or II defined above.

8. A composition according to any one of Claims 4 to 7 in which the propellant is selected from Cl4alkane and carbon dioxide.

9. A composition according to any one of Claims 4 to 8 in which the solvent is selected from C16 alcohols (e.g. ethanol) or water.

10. A composition according to any one of Claims 4 to 9 in which the composition may also contain one or more of the following a fragrance, a surfactant (e.g. Dow Corning 193 Surfactant an antimicrobial agent (e.g.alkyl dimethyl benzyl ammonium saccharinate), a corrosion inhibitor (e.g. sodium nitrite, sodium benzoate, triethanolamine and ammonium hydroxide), and/or a miticide (such as benzyl benzoate).

11. A method for denaturing a Der-f allergen substantially as herein described with reference to any one of the Examples.

12. A composition substantially as herein described with reference to any one of the Examples.