The present invention relates to novel protein-cyclodextrin derivatives useful in the preparation of hair care and skin care compositions. Compounds according to the present invention are particularly useful in systems for extending fragrance retention and de-odorising applications.

Hair care and skin care compositions often contain fragrance components. By their nature these components are volatile leading to them having short retention times, therefore extra fragrance is often included in a formulation to account for this. Fragrance components are some of the most expensive components found in such formulations and so a method of prolonging fragrance retention would be valuable in prolonging the effect of the composition as well as allowing less of the fragrance material to be included in a formulation.

Hair and skin can also develop unpleasant odours by uptake from external environmental sources such as cigarette smoke and cooking smells and from internal sources such as body odours, thus imparting an unpleasant odour to hair and skin. Reducing or eliminating the sensory detection of these malodours is highly desirable.

The term "body odour" or "body malodour", as used herein, means odours that are generated as a result of the natural functioning of a human or mammalian body. Such odours include, but are not limited to, odours produced by microorganisms of the human or mammalian skin (i.e. bacterial decomposition of skin secretions) or urine, and mixtures thereof. Such odours are mainly organic molecules, which have different structures and functional groups, such as amines, acids, alcohols, aldehydes, ketones, phenolics, and polycyclics including aromatics and, polyaromatics. These molecules may also be made up of sulphurs containing functional groups, such as thiol, mercaptan, sulphide and/or disulphide groups.

Cyclodextrins are ring shaped molecules, built up from glucose units [D-(+)- glucopyranose units]. Cyclodextrins occur in nature and can be produced commercially by enzymatic transformation of starch. The 3 dimensional structure of cyclodextrin molecules leads to the formation of a cavity in the centre of the molecule that can trap a 'guest1 molecule. The size of the cavity depends on the number of glucose units in the ring structure. The most common forms of cyclodextrin have 6, 7 and 8 glucose units and are called alpha-, beta- and gamma- cyclodextrin respectively. Cyclodextrin will form inclusion complexes with certain materials. Inclusion complexes occur when a guest molecule becomes incorporated into the cavity of the cyclodextrin. This ability to form inclusion complexes has led to a number of common applications for cyclodextrins.

One such application for cyclodextrins is application to substrates / surfaces with the intention of removing malodors. There are many publications concerning this application of which only a few examples follow. World Patent Application No WO 2003020231 describes a composition containing cyclodextrin for use as a skin deodorant. US patent specification No US6,284,231 describes a spray formulation for use as an odour absorbing composition on inanimate surfaces. World patent specification No WO9,856,340 describes powdered compositions containing cyclodextrin-encapsulated perfume for use in body odour control compositions. US patent specification No US6,436,442 describes methods for manufacturing compositions comprising cyclodextrin. This patent is concerned with a method of formulating around the problems normally associated with cyclodextrins. The application is for capturing unwanted molecules, malodours, from inanimate and animate surfaces such as skin and hair. Reference is also made to helping the effects of fragrance in the formulation to endure. The cyclodextrins preferred are the more water-soluble alpha- and gamma- cyclodextrins and beta-cyclodextrins modified to enhance water solubility.

WO9,921 ,532 discloses "a substantially dry, disposable, personal cleansing product useful for both cleansing and conditioning the skin/hair and providing improved fragrance delivery". The composition uses cyclodextrin and modified cyclodextrin as one example of a compound that can aid fragrance delivery in this application.

A further application of cyclodextrins is to help improve solubility, stability and availability of active compounds. US patent specification No 6,080,733 describes modified cyclodextrins containing a thioureido linking group. The modified cyclodextrin is used to solubilise anti-tumour and anti-parasitic drugs in aqueous media. US Patent Application No US 20030144222 describes the grafting of cyclodextrin onto a biocompatible amphiphilic polymer. The purpose of the invention is to modify the cyclodextrin properties to improve water solubility of the complexed cyclodextrin, thus encouraging tissue penetration of active agents complexed with the cyclodextrin in both cosmetic and dermopharmaceutical applications.

US Patent Application No US5,324,750 describes the modification of cyclodextrins to improve solubility and stability of inclusion complexes by covalently linking active compounds to cyclodextrin for use in orally delivered compositions. The covalently linked active compounds will then be released after the cyclodextrin molecule has been metabolised to glucose.

US patent specification no. US 4 678 598 specifies the use of a methylated cyclodextrin in a shampoo formulation. The purpose of the cyclodextrin is to mitigate the strong odour of the skin sensation inducing aromatic chemical that is being applied by use of the shampoo.

None of the above prior art considers the substantivity, if any, that cyclodextrins, or compounds containing cyclodextrin, may have for the skin or the hair.

Hydrolysed proteins are common personal care ingredients that are used as conditioning and moisturising agents for skin and hair. Hydrolysed proteins are known to have substantivity to hair from aqueous systems as described in the article The use of radiolabelling techniques to measure substantivity to, and penetration into, hair of protein hydrolysates' by RT Jones and S. P. Chahal, International Journal of Cosmetic Science, 1997, Vo1 19, pages 215-226.

The present invention retains the moisturising and conditioning properties of the hydrolysed protein component and the fragrance controlled release and malodour control properties of the cyclodextrin component. The hydrolysed protein component imparts substantivity to the protein cyclodextrin component, an unexpected advantage over the un-modified cyclodextrin component. Accordingly, the present invention provides protein-cyclodextrin derivatives obtained by the reaction of a reactive cyclodextrin and a protein, and their use in a variety of cosmetic compositions.

Preferred protein-cyclodextrin derivatives have the general formula (I):

[R'-(NH)z]w- (Tb-C)p (I)

wherein R'-(NH) is the residue of a protein R'-NH2; T is the residue of an organofunctional group attached to cyclodextrin; C is cyclodextrin; b corresponds to the number of organofunctional groups on the reactive cyclodextrin and is preferably in the range 1 to 8; p corresponds to the number of organofunctional cyclodextrin reacted with amino groups on protein residues and will be preferably in the range of 1 to (w x z); 2 is the number of primary amine groups on the protein R'-NH2 available for reaction with the organofunctional cyclodextrin; w is the number of protein residues and is preferably in the range of 1 to (p x b).

Preferably R1-(NH) is the residue of a protein, more preferably a hydrolysed protein and even more preferably a vegetable protein hydrolysate such as a hydrolysate derived from potato, wheat, soya or brazil nut protein, and preferably potato protein. The protein residue can be modified and can also be an amino acid or mixture of amino acids. In the case of an amino acid, the amino acid can either be derived from a protein or made synthetically. In the latter case, this may not necessarily be by hydrolysis.

In a preferred embodiment a protein and a reactive cyclodextrin are reacted in the range of 50% to 200% reactive cyclodextrin by weight of active protein. A further preferred embodiment provides a protein-cyclodextrin derivative having the general formula (III):

wherein:

and R1-NH is the residue of a protein, preferably a hydrolysed protein.

In further aspects the invention also provides, separately, for the use of protein- cyclodextrin derivatives as fragrance delivery agents, fragrance retention agents and malodour control agents. The present invention therefore provides a protein-cyclodextrin derivative obtainable by reacting a protein and a reactive cyclodextrin compound in the range from 10% to 500% reactive cyclodextrin by weight of active protein, more preferably 50 to 200% reactive cyclodextrin by weight of active protein. The reactive groups on the protein component are predominantly primary amino groups. The primary amino groups are either located at the end of the polypeptide chains of the protein or as side chain groups on basic amino acids such as lysine. The degree of hydrolysis of the protein component will affect the number of end chain primary amino groups available for reaction. The greater the degree of hydrolysis of the protein component, the higher the number of end chain primary amino groups available for reaction. Therefore the degree of hydrolysis of the protein component will determine the number of reactive sites on the protein component available for derivatisation with the reactive cyclodextrin. Thus the ratio of protein to cyclodextrin in the finished product will vary. Similarly the reactive cyclodextrin compound can have more than one group available for reaction with the protein component of the product. This can also affect the ratio of protein to cyclodextrin in the final product. Due to the possibility of multiple reaction sites on both the protein and cyclodextrin components of the present invention, the present invention could comprise: (i) one protein molecule reacted with one cyclodextrin molecule, (ii) more than one protein molecule reacted with one cyclodextrin molecule, (iii) one protein molecule reacted with more than one cyclodextrin molecule, (iv) more than one protein molecule reacted with more than one cyclodextrin molecule, i.e. a co-polymeric or cross-linked product, (v) a combination of examples (i) to (iv).

The present invention also provides the use of a protein-cyclodextrin derivative composition to prolong the fragrance perceptibility of a formulation or aqueous solution applied to skin or hair.

The terms "fragrance" and "perfume" are intended to have the same meaning in this disclosure. The fragrance or perfume is included at a level that is non-irritating to the ordinary user's skin and/or respiratory tract, yet is discernible by the human sense of smell either before and/or after application to the skin. The perfume compositions should be safe for use on skin. The perfume compositions useful herein are comprised of perfume ingredients. Perfume loading in the cyclodextrin complexes is typically from about 0.1% to about 20% in cyclodextrin/perfume complexes. The cyclodextrin. perfume ingredient inclusion complexes and methods of formulation useful herein, are disclosed in detail in U.S. Patent No. 5,429,628, Trinh et al. issued July 4, 1995, which is incorporated herein by reference in its entirety.

A wide variety of perfume ingredients can be used in the present invention and such perfume ingredients will be selected by the person skilled in the art. A non-limiting list of preferred volatile perfume ingredients can be found in WO 98/56340 (Petersson et al) the entire text of which is incorporated herein by reference and which is intended to be an integral part of this disclosure.

It will be appreciated that the cyclodextrin derivative can be used to complex "actives" other than a perfume or fragrance. Thus these derivatives can be used to deliver to the skin or hair active agents such as antifungal agents, anti-dandruff agents, antibacterials, antibiotics, UK protective agents or the like. It is therefore possible, for the first time, to create a long lasting anti-dandruff shampoo or conditioner as a result of the slow release of active agent directly onto the hair. Furthermore, as the active ingredient is released the emptied cyclodextrin moieties become available to absorb any malodour which might occur in the course of the users normal activities.

It is possible that a combination of compounds could be complexed with cyclodextrin. For example a mixture of perfume ingredients and an antifungal agent could be used.

The present invention provides the use of a protein-cyclodextrin derivative composition to control malodour when applied from a formulation or aqueous solution to skin or hair.

In order to form the protein cyclodextrin derivative of the present invention the protein and reactive cyclodextrin are reacted in the range from 10% to 500% reactive cyclodextrin by weight of active protein, preferably 50% to 200% reactive cyclodextrin by weight of active protein. The resulting derivative comprises 33% to 66% cyclodextrin and modified reactive amino groups of the protein in the range 10 - 100%. We have found that hair treated with protein cyclodextrin derivatives of the present invention retains perceptible fragrance applied from a formulation for a significantly longer period than formulations without a protein cyclodextrin component.

We have found that hair treated with protein cyclodextrin derivatives of the present invention when exposed to malodours, for example environmental malodours, exhibit a reduction in perceptible malodour. One example of this is tobacco smoke were it has been found that hair treated with protein cyclodextrin derivatives of the present invention reduce the perceptible tobacco malodour.

Preferred derivatives for use according to the invention are those wherein the protein cyclodextrin derivative is the reaction product of a protein hydrolysate and a reactive cyclodextrin. More preferably the reactive cyclodextrin contains a chloro- triazine group capable of reacting with one or more amino groups of the protein, such as monochlorotriazinyl-β-cyclodextrin.

The protein may be derived from either animal or vegetable sources or by fermentation. It may be in the form of a chemically modified protein (for example, quatemised, silanised or copolymerised) provided that at least one free amino groups is still present in the protein molecule. Examples of proteins which are currently used in cosmetic formulations and can be used as the protein component of the current invention include collagen (including bovine, porcine marine and avian), elastin (including bovine, porcine and marine), casein (milk and whey), cereal (including wheat, soya, maize [corn], oat), rice, pea, proteins from seeds or nuts (eg., brazil nut, sesame, cotton, apricot etc), algal, keratin (including hoof and horn, wool, human hair, etc.), silk, egg and potato. In this specification the term "protein" is used to include both native and hydrolysed proteins and it thus comprises both proteins properly so-called polypeptides, peptides, peptones and amino acids, since the latter can all be categorised as hydrolysed proteins.

The present invention encompasses any residue derived from or found in a protein. It is possible to use amino acids as the protein residue. These can be derived from a protein, for example by hydrolysis, or can alternatively be synthetically made. It should be understood that the term "protein" or "residue of a protein" also encompasses amino acids, whether made synthetically or whether derived from a protein by hydrolysis or by some other means. For the avoidance of doubt, the term amino acid also encompasses modified amino acids or an amino acid backbone with different or modified side chains.

All amino acids are encompassed by the present invention. The present invention also covers mixtures of amino acids. Preferably, the amino acids can be selected from arginine, lysine or cystine. Arginine and lysine are cationic and therefore have better substantivity. Cystine has a disulphide bridge a and is therefore capable of reacting with the disulphide bridges in hair for better retention.

The average molecular weight of the protein component may be from 75 Daltons (D) to 500 kD, is preferably within the range from 75D to 5OkD and is more preferably in the range of from 75D to 25.5kD, even more preferably 75D to 3000D and particularly preferably 350D to 3000D expressed as weight average molecular weight (Mw) derived from size exclusion chromatography.

Especially preferred is when the protein hydrolysate is a vegetable protein hydrolysate, particularly wherein the vegetable protein hydrolysate is of potato, wheat or brazil nut origin.

It is necessary for the protein component to be capable of solution in water or other suitable solvent or co-solvent (such as alcohol, eg propylene glycol or polyethylene glycol) to enable reaction to occur.

The generic term "cyclodextrin" in the present invention is intended to include unsubstituted and substituted cyclodextrins both known and yet to be discovered. Cyclodextrins suitable for use in the present invention include any of the known cyclodextrins such as unsubstituted cyclodextrins containing from 6 to 12 glucose units. Specific nonlimiting examples of such cyclodextrins include alpha- cyclodextrin, beta-cyclodextrin, gamma-cyclodextrin, delta-cyclodextrin, epsilon- cyclodextrin, zeta-cyclodextrin, nu-cyclodextrin, and mixtures thereof, and/or their derivatives, and/or mixtures thereof.

Suitable cyclodextrin derivatives include those cyclodextrin compounds of different degrees of substitution, specific examples of which include methyl-alpha- cyclodextrin, methyl-beta-cyclodextrin, hydroxyethyl-beta-cyclodextrin, hydroxypropyl-alpha-cyclodextrin, hydroxypropyl-beta-cyclodextrin, cyclodextrin glycerol ethers, maltose-bonded cyclodextrins, cationic cyclodextrins, quarternary ammonium cyclodextrins, anionic cyclodextrins such as carboxymethyl cyclodextrins, cyclodextrin sulfobutylethers, cyclodextrin sulphates, cyclodextrin succinylates, amphoteric cyclodextrins such as carboxymethyl/quatemary ammonium cyclodextrins, mono-3-6-anhydrocyclodextrins, and combinations thereof. Other examples of suitable cyclodextrin derivatives are disclosed in Optimal Performances with Minimal Chemical Modification of Cyclodextrins", F Diedaini-Pilard and B. Perly, The 7th International Cyclodextrin Symposium Abstracts, April 1994, p.49; U.S. Patent No. 3,426,011 , issued to Parmerter et al. on Feb.4, 1969; U.S. Patent Nos. 3,453,257, 3,453,258, 3,453,259 and 3,453,260, all issued to Parmerter et al. on Aug.5, 1969; U.S. Patent No. 3,553,191 , issued to Parmerter et al. on Jan.5, 1971 ; U.S. Patent No. 3,565,887, issued to Parmerter et al. on Feb 23, 1971 ; U.S. Patent No. 4,535,152, issued to Szejtli et al. on Aug.13, 1865; U.S. Patent No. 4,616,008, issued to Hirai et al. on Oct.7, 1986; U.S. Patent No. 4,638,058, issued to Brandt et al. on Jan.20, 1987; U.S. Patent No. 4,746,734, issued to Tzuchiyama et al. on May 24, 1988; and U.S. Patent No. 4,678,598, issued to Ogino et al. on JuI.7, 1987; all of which disclosures are incorporated by reference herein. This list is not intended to be limiting. Rather it illustrates the very large range of cyclodextrins both known and yet to be discovered which can be used in the present invention.

It is also possible to use a mixture of cyclodextrins. Such mixtures absorb body odours more broadly by complexing with a wider range of odoriferous molecules having a wider range of molecular sizes.

The term "uncomplexed cyclodextrin" as used herein means that the cavities within the cyclodextrin in the composition are essentially unfilled.

The organofunctional cyclodextrin component is preferably soluble in a solvent common to the protein for efficient reaction to occur. The conditions for reaction of the organofunctional cyclodextrin with the protein to achieve efficient substitution of primary amine groups on the protein must be carefully controlled, but will be apparent or readily determinable by those skilled in the art of protein modification. As explained above, the organofunctional cyclodextrin component may be any of the known cyclodextrins containing from six to twelve glucose units, especially alpha- cyclodextrin, beta-cyclodextrin or gamma-cyclodextrin cyclodextrin, more especially beta-cyclodextrin. The alpha cyclodextrin consists of six glucose units, beta cyclodextrin consists of seven glucose units, gamma cyclodextrin consists of eight glucose units.

The organofunctional cyclodextrin component used for reaction with the protein component must contain at least one functional group capable of reacting with the chain terminal and/or side chain amino groups of the protein. The cyclodextrin component can contain more than one functional group capable of reacting with the chain terminal and/or side chain amino groups of the protein. Suitable reactive groups include, for example, chloro-triazinyl, epoxide, acyl halide, sulphonyl halide, anhydride, and aldehyde groups. One particular example of a reactive cyclodextrin is Cavasol W7 MCT produced by Wacker Chemie. This product comprises beta cyclodextrin containing chloro-triazinyl reactive groups.

A suitable method for the manufacture of protein / cyclodextrin compositions for use according to the invention is represented by the following steps:

In a suitable vessel, the protein component is heated and the pH adjusted to ensure the conditions favour the reaction of the cyclodextrin functional group with amine groups on the protein over reaction with water in a hydrolysis reaction. A calculated quantity of organofunctional cyclodextrin is then added - the amount of cyclodextrin is calculated to modify 10-100%, preferably 50-90% of the available amino groups. An excess of a primary amine containing compound, such as an amino acid, can then be added to ensure that all of the reactive groups on the organofunctional cyclodextrin have reacted. Following reaction, the pH is adjusted to acidic and worked up in a conventional manner. Conventional additives, such as preservatives, may then be added.

A preferred method for the manufacture of protein / cyclodextrin compositions for use according to the invention is represented by the following steps:

(1) Load the protein component, preferably hydrolysed vegetable protein, into a suitable tank and heat to 20 - 9O0C (preferably 60 - 8O0C). (2) Adjust pH to 6.0 - 10.0 (preferably 7.0-9.0) with sodium hydroxide. (3) Add calculated quantity of reactive cyclodextrin (this is based on Formol titre) and is calculated to modify 10 - 100% (preferably 80 - 90%) of the available amino groups, over a 30 to 120 minute period (preferably 45 to 75 minute period). (4) Maintain temperature at 20-900C (preferably 60-800C) and pH at 6.0 - 10.0 (preferably 7.0 - 9.0) during reactive cyclodextrin addition. (5) Maintain reaction conditions for a further 4 to 20 hours (preferably 7 - 10 hours). (6) Add 0.5% - 4% (preferably 1%-3%) of an amino acid (optional) such as glycine and maintain temperature at 20-900C (preferably 60-800C) and pH at 6.0 - 10.0 (preferably 7.0 - 9.0) for 2 to 10 hours (preferably 4 to 8 hours). (7) Acidify to pH 4.0 - 6.5 with a mineral acid such as hydrochloric acid. (8) Preserve by addition of a preservative. (9) Cool and adjust pH to 4.0 - 6.5. (10) Filter through a depth filter pad to sparkling.

Accordingly an aspect of the present invention comprises a compound of general formula (I):

[R'-(NH)z]w- (Tb-C)p (I)

wherein R'-(NH) is the residue of a protein R'-NH2; T is the residue of an organofunctional group attached to cyclodextrin; C is cyclodextrin; b corresponds to the number of organofunctional groups on the reactive cyclodextrin and will be preferably in the range 1 to 8; p corresponds to the number of organofunctional cyclodextrin reacted with amino groups on protein residues and will be preferably in the range of 1 to (w x z); z is the number of primary amine groups on the protein R'-NH2 available for reaction with the organofunctinal cyclodextrin; w is the number of protein residues and will preferably be in the range of 1 to (p x b).

R' is preferably the residue of a hydrolysed protein, more preferably a hydrolysed vegetable protein, such as hydrolysed wheat, potato or brazil nut protein, especially potato protein. The nature of the group T, which is the residue of the organofunctional group of the reactive cyclodextrin used for reaction with the protein component to form the derivative, depends on the functional group originally present in the organofunctional cyclodextrin. For example, when the functional group originally present in the organofunctional cyclodextrin was an acyl halide, the group T will typically contain a carbonyl group bonded directly to the nitrogen atom of the protein residue. Preferably the group T is derived from reaction of the organofunctional cyclodextrin containing chloro-triazine groups as the functional group. In this case, in the final derivative of compound (I), the group T will contain a moiety of formula (II), with the nitrogen atom of the protein residue, R' bonded directly to one of the cyclic carbon atoms. The group C in formula (II) represents cyclodextrin. The group Y in formula (II) represents the residue of the compound used to remove the chloride group from this site that was present in cyanuric chloride from which this linking group is generally derived. In this case Y can be, but is not limited to OH, NH2, NHP, NPP', where P and P' are substituents on secondary and tertiary amines. Preferably this group, Y, is OH or an OH group deprotonated to O".

The protein-cyclodextrin derivative preferably comprises a compound of formula (III) where R' is the residue of a protein, preferably a hydrolysed protein; the groups R indicate that there may be more than one protein residue attached to each cyclodextrin molecule: (III)

In the protein cyclodextrin derivative for use in the invention, carboxyl groups of the protein component and/or amino groups of the protein which are not bonded to the cyclodextrin component, can be chemically modified by reaction with a non- cyclodextrin, eg by esterification, acylation or quatemisation.

The derivatives of the present invention may be used to prepare simple aqueous compositions for application to the hair or skin, such as an aqueous "leave on" composition or an aqueous "rinse off' composition.

For such compositions, a dilute solution of the derivative in water may be used. The concentration of active ingredient in such a solution may range from 0.01 % w/w to 10% w/w and is preferably 0.05% w/w to 5% w/w, more preferably 0.5% w/w to 2% w/w and most preferably about 1% w/w.

Compounds and compositions according to the present invention can be also used in a wide variety of skin care formulations since hydrolysed proteins have substantivity for skin as well as hair. Skin care compositions include, but are in no way limited to, cleansing preparations, skin toners, moisturisers, scrubs, masks including face masks, washes, wipes, soap bars, creams, astringents, anti-ageing creams, anti-wrinkle creams, solar creams and tanning products, lip balms, make-up removers, exfoliants, perfumes, shower gels, bath oils, lotions and the like.

The present invention will now be illustrated with reference to the following examples.

Example 1 : Preparation of hydrolysed protein cyclodextrin derivative The following steps were carried out: 1. Hydrosolanum (100Og), a potato protein hydrolysate available from Croda Chemicals Europe Ltd, was loaded into a suitable beaker and stirred. The solution was heated to 750C. 2. The pH was adjusted to 7.5 using 25% sodium hydroxide (68.8g) 3. Cavasol W7 MCT (313.5g), a compound of formula (A) below, (available from Wacker-Chemie GmbH, Hanns-Seidel-Platz 4, DE-81737 Munchen, Germany) was added over 60 minutes. 4. During addition, pH was maintained in the range 7.0-7.5 using 25% sodium hydroxide (71.5g) and the temperature kept constant at 750C. 5. The reaction mixture was stirred for a further 8 hours at 750C. 6. 28% Hydrochloric acid (53.6g) was added to lower the pH to 5.5. 7. Phenoxyethanol (14.4g), potassium sorbate (3.6g) and disodium EDTA (1 -8g) were added and the solution stirred overnight at room temperature. 8. The liquor was then filtered through a depth filter pad to sparkling.

(A)

Example 2: Preparation of quaternised hydrolysed protein cyclodextrin derivative The following steps were carried out: 1. Hydrosolanum (50Og) (Croda Chemicals Europe Ltd), was loaded into a suitable beaker and stirred. The solution was heated to 750C. 2. The pH was adjusted to 7.5 using 25% sodium hydroxide (35.1g) 3. Cavasol W7 MCT (78.3g), (available from Wacker-Chemie GmbH, Hanns- Seidel-Platz 4, DE-81737 Munchen, Germany) was added over 60 minutes. 4. During addition, pH was maintained in the range 7.0-7.5 using 25% sodium hydroxide (18.Og) and the temperature kept constant at 750C. 5. The reaction mixture was stirred for a further 6 hours at 750C. 6. The reaction mixture was cooled to 6O0C and held at this temperature. 7. The pH of the reaction mixture was raised to 10.0 to 10.2 using 25% sodium hydroxide (22.5g). 8. 3-chloro-2-hydroxypropyltrimethyl ammonium chloride (Quab 151 , available from Degussa) (31.8g) was added dropwise via a separating funnel over a one hour period with the pH of the reaction mixture maintained at 10.0 to 10.2 using 25% sodium hydroxide. 9. The reaction mixture was stirred at 6O0C, with the pH maintained at 10.0 to 10.2 using 25% sodium hydroxide (2.9g), for 4 hours. 10. 28% Hydrochloric acid (42.6g) was added to lower the pH to 4.5. 11. Phenoxyethanol (6.4g), potassium sorbate (1.6g) and disodium EDTA (0.8g) were added and the solution stirred overnight at room temperature. 12. The liquor was then filtered through a depth filter pad to sparkling.

Example 3: Measurement of fragrance retention of a protein cyclodextrin derivative according to Example 1 applied to hair from an aqueous solution Aqueous test solutions containing the fragrance citral (a mixture of cis- and trans- 3,7-dimethyl-2,6-octadienal), have been applied to hair swatches. An aqueous citral solution containing hydrolysed protein cyclodextrin derivative has been applied to hair from a rinse off system. The fragrance retention performance of the aqueous citral solution containing hydrolysed protein cyclodextrin derivative (as in example 1) has been assessed against an aqueous citral solution containing no fragrance retention aids. Materials used in example 3 Virgin European hair (de Meo) 10% Sodium laureth sulphate (SLES) Aqueous formulation without active Aqueous formulation with hydrolysed protein cyclodextrin derivative (as in example 1) (1.0% active)

Aqueous test formulation % by Wt Water up to 100 Crillet 1 (Croda Chemicals Europe Ltd) 4.0 Citral (Sigma-Aldrich Chemical Company) 0.4 Hydrolysed protein cyclodextrin derivative qs (to provide 1.0% of active)

Study procedure 1. Virgin brown European hair was cut into swatches (300mg) and tied with cotton. The swatches were labelled A and B. 2. Each swatch was soaked in the aqueous test solution for 1 minute. 3. The A swatch was soaked in the aqueous test solution without active for 1 minute. 4. The B swatch was soaked in the aqueous test solution containing hydrolysed protein cyclodextrin derivative (1.0% active) for 1 minute. 5. The hair swatches were removed from the aqueous test solutions and rinsed under a running cold tap for 10 seconds. 6. The hair swatches were blotted dry on tissues. 7. The swatches were allowed to dry at ambient temperature and relative humidity for the desired time period (1 , 2, or 6 hours) 8. The swatches were transferred to separate 5ml test tubes and methanol (HPLC grade, 5ml) added. 9. The test tubes were covered and the hair left for 30 minutes. 10. The hair swatches were removed and the methanol extracts analysed by Gas Chromatography under the following conditions: Gas Chromatography analysis conditions Instrumentation: Agilent 6850 Series GC, with Agilent 7683 series injector Software: Agilent Chemstation software Injector: Injection volume: 5μl Splitless injection Temperature: 22O0C Pressure: 44.2kPa Total Flow: 138ml/min Purge flow to split vent: 130ml/min at 0.5min Gas: Hydrogen Column: Agilent HP1 Methyl Siloxane Constant pressure mode H2 pressure: 44.3 kPa H2 flow: 2.8 ml/min Column average velocity: 50cm/s Oven: Initial temperature: 5O0C Initial hold time: 1 min Ramp 1 : 50-1100C, 20°C/min Ramp 2: 110-1400C, 5°C/min Ramp 3: 140-3000C, 40°C/min Final hold time: 7 minutes Total run time: 21 minutes Flame ionisation detector

Citral shows two peaks on GC analysis (due to cis and trans isomers). The amount of citral present in the methanol extracts was determined from the ratio of the peak area detected for each peak to the peak area obtained for a sample of citral in methanol of known concentration. Fragrance retention values were calculated for each peak at each drying time using the following equation:

Percentage fragrance retention = A0 x 100 Where: A0 = Average peak area after 1 hour of drying At = Average peak area after 1 , 2, 6 hours of drying

The fragrance retention values for the two peaks obtained at each drying time were averaged. The results are shown in table 1 , with standard deviation (SD) values for each data point, and in figure 1.

Table 1 : % fragrance retention results for hair swatches treated with hvdrolvsed protein cvclodextrin derivative and control

The study demonstrates that the hair samples treated with aqueous fragrance solutions containing hydrolysed protein cyclodextrin derivative retained the fragrance, citral, to a greater extent than hair treated with the aqueous fragrance solution alone. Figure 1 : % fragrance retention results for hair swatches treated with hvdrolvsed protein cvclodextrin derivative and control

(as in

Time (hours)

Example 4: Measurement of fragrance retention of a quaternised protein cydodextrin derivative according to Example 2 applied to hair from an aqueous solution Aqueous test solutions containing the fragrance, citral (a mixture of cis- and trans- 3,7-dimethyl-2,6-octadienal), have been applied to hair swatches. An aqueous citral solutions containing quaternised hydrolysed protein cydodextrin derivative has been applied to hair from a rinse off system. The fragrance retention performance of the aqueous citral solution containing quaternised hydrolysed protein cydodextrin derivative (as in example 2) has been assessed against an aqueous citral solution containing no fragrance retention aids.

Materials used in example 4 Virgin European hair (de Meo) 10% Sodium laureth sulphate (SLES) Aqueous formulation without active Aqueous formulation with quaternised hydrolysed protein cydodextrin derivative (as in example 2) (1.0% active) Aqueous test formulation % by Wt Water up to 100 Crillet 1 (Croda Chemicals Europe Ltd) 4.0 Citral (Sigma-Aldrich Chemical Company) 0.4 Quatemised Hydrolysed protein qs (to provide 1.0% of active) cyclodextrin derivative

Study procedure 1. Virgin brown European hair was cut into swatches (300mg) and tied with cotton. The swatches were labelled A and B. 2. Each swatch was soaked in the aqueous test solution for 1 minute. 3. The A swatch was soaked in the aqueous test solution without active for 1 minute. 4. The B swatch was soaked in the aqueous test solution containing quatemised hydrolysed protein cyclodextrin derivative (1.0% active) for 1 minute. 5. The hair swatches were removed from the aqueous test solutions and rinsed under a running cold tap for 10 seconds. 6. The hair swatches were blotted dry on tissues. 7. The swatches were allowed to dry at ambient temperature and relative humidity for the desired time period (1 , 2, or 6 hours) 8. The swatches were transferred to separate 5ml test tubes and methanol (HPLC grade, 5ml) added. 9. The test tubes were covered and the hair left for 30 minutes. 10. The hair swatches were removed and the methanol extracts analysed by Gas Chromatography under the following conditions:

Gas Chromatography analysis conditions As in example 3.

Citral shows two peaks on GC analysis (due to cis and trans isomers being present). The amount of citral present in the methanol extracts was determined from the ratio of the peak area detected for each peak to the peak area obtained for a sample of citral in methanol of known concentration. Fragrance retention values were calculated for each peak at each drying time using the following equation: Percentage fragrance retention = A0 x 100 "AT

Where: A0 = Average peak area after 1 hour of drying At = Average peak area after 1, 2, 6 hours of drying

The fragrance retention values for the two peaks obtained at each drying time were averaged. The results are shown in table 2, with standard deviation (SD) values for each data point, and in figure 2.

Table 2: % fragrance retention results for hair swatches αuaternised hvdrolvsed protein cvclodextrin derivative and control

The study demonstrates that the hair samples treated with aqueous fragrance solutions containing quatemised hydrolysed cyclodextrin derivative retained the fragrance, citral, to a greater extent than hair treated with the aqueous fragrance solution alone. Figure 2: % fragrance retention results for hair swatches treated with quaternised hvdrolvsed protein cvclodextrin derivative and control

cyclodextπn

4 Time (hours)

Example 5: Measurement of fragrance delivery of a quaternised protein cyclodextrin derivative according to Example 2 applied to hair from a shampoo system Shampoo test solutions containing the fragrance, citral (a mixture of cis- and trans- 3,7-dimethyl-2,6-octadienal), have been applied to hair swatches. A shampoo containing citral and quaternised hydrolysed protein cyclodextrin derivative has been applied to hair from a rinse off system. The fragrance delivery performance of the shampoo containing quaternised hydrolysed protein cyclodextrin derivative (as in example 2) has been assessed against a shampoo formulation containing no quaternised hydrolysed protein cyclodextrin derivative.

Materials used in example 5 Virgin European hair (de Meo) Shampoo formulation without quaternised hydrolysed protein cyclodextrin derivative Shampoo formulation with quaternised hydrolysed protein cyclodextrin derivative (as in example 2) (1.0% active) Shampoo formulation % by Wt Water up to 100 Empicol ESB3 (Allbright & Wilson) 35.0 lncronam 30 (Croda Chemicals Europe Ltd) 15.0 Crovol A70 (Croda Chemicals Europe Ltd) 3.0 Lactic acid (Sigma-Aldrich Chemical Company) to pH 6.5 - 7.0 Citral (Sigma-Aldrich Chemical Company) 0.4 Phenova (Crodarom) 0.4 Hydrolysed quaternised protein qs (to provide 1.0% of active) cyclodextrin derivative

Study procedure 1. Virgin brown European hair was cut into swatches (300mg) and tied with cotton. The swatches were labelled A and B. 2. Each swatch was wet in running cold water for 10 seconds. 3. 5ml of shampoo formulation without hydrolysed quaternised protein cyclodextrin was massaged into the A swatch for 1 minute. 4. 5ml of shampoo formulation without hydrolysed quaternised protein cyclodextrin was massaged into the B swatch for 1 minute. 5. The hair swatches were rinsed under a running cold tap for 10 seconds. 6. The hair swatches were blotted dry on tissues. 7. The swatches were allowed to dry at ambient temperature and relative humidity for one hour. 8. The swatches were transferred to separate 5ml test tubes and methanol (HPLC grade, 5ml) added. 9. The test tubes were covered and the hair left for 30 minutes. 10. The hair swatches were removed and the methanol extracts analysed by Gas Chromatography under the following conditions:

Gas Chromatography analysis conditions As in example 3.

Citral shows two peaks on GC analysis (due to cis and trans isomers). The amount of citral present in the methanol extracts was determined from the ratio of the peak area detected for each peak to the peak area obtained for a sample of citral in methanol of known concentration. Fragrance delivery values were calculated for each peak using the following equation:

Percentage fragrance delivery = Weight of citral in the methanol extract x 100 Weight of citral in 5ml shampoo

The fragrance delivery values for the two peaks obtained were averaged. The results are shown in table 3, with standard deviation (SD) values for each data point, and in figure 3.

Table 3: % fragrance delivery results for hair swatches treated with a shampoo containing quaternised hvdrolvsed protein cvclodextrin derivative and control

% fragrance delivery for a shampoo containing 0.93 quaternised hydrolysed protein cyclodextrin (0.29)

% fragrance delivery for a shampoo without quaternised 0.59 hydrolysed protein cyclodextrin (0.19)

Figure 3: % fragrance delivery results for hair swatches treated with guaternised hvdrolvsed protein cvclodextrin derivative and control

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The study demonstrates that the shampoo containing quatemised hydrolysed protein cyclodextrin derivative delivered the fragrance, citral, to hair to a greater extent than shampoo without quatemised hydrolysed protein cyclodextrin derivative.

Example 6: Measurement of fragrance delivery of a quatemised protein cyclodextrin derivative according to Example 2 applied to hair from a conditioner system Conditioner test solutions containing the fragrance citral (a mixture of cis- and trans- 3,7-dimethyl-2,6-octadienal), have been applied to hair swatches. A conditioner containing citral and quatemised hydrolysed protein cyclodextrin derivative has been applied to hair from a rinse off system. The fragrance delivery performance of the conditioner containing quatemised hydrolysed protein cyclodextrin derivative (as in example 2) has been assessed against a conditioner formulation containing no quatemised hydrolysed protein cyclodextrin derivative.

Materials used in example 6 Virgin European hair (de Meo) Conditioner formulation without quatemised hydrolysed protein cyclodextrin derivative Conditioner formulation with quatemised hydrolysed protein cyclodextrin derivative (as in example 2) (1.0% active)

Conditioner formulation % by Wt Water up to 100 Incroquat CTC-30 (Croda Chemicals Europe Ltd) 10.0 Crodacol CS90 (Croda Chemicals Europe Ltd) 5.0 Crillet 1 (Croda Chemicals Europe Ltd) 3.2 Citral (Sigma-Aldrich Chemical Company) 0.4 Phenova (Crodarom) 0.4 Hydrolysed quatemised protein qs (to provide 1.0% of active) cyclodextrin derivative Study procedure 1. Virgin brown European hair was cut into swatches (300mg) and tied with cotton. The swatches were labelled A and B. 2. Each swatch was wet in running cold water for 10 seconds. 3. 5ml of conditioner formulation without hydrolysed quatemised protein cyclodextrin was massaged into the A swatch for1 minute. 4. 5ml of conditioner formulation without hydrolysed quatemised protein cyclodextrin was massaged into the B swatch for1 minute. 5. The hair swatches were rinsed under a running cold tap for 10 seconds. 6. The hair swatches were blotted dry on tissues. 7. The swatches were allowed to dry at ambient temperature and relative humidity for one hour. 8. The swatches were transferred to separate 5ml test tubes and methanol (HPLC grade, 5ml) added. 9. The test tubes were covered and the hair left for 30 minutes. 10. The hair swatches were removed and the methanol extracts analysed by Gas Chromatography under the following conditions:

Gas Chromatography analysis conditions As in example 3.

Citral shows two peaks on GC analysis (due to cis and trans isomers). The amount of citral present in the methanol extracts was determined from the ratio of the peak area detected for each peak to the peak area obtained for a sample of citral in methanol of known concentration. Fragrance delivery values were calculated for each peak using the following equation:

Percentage fragrance delivery = Weight of citral in the methanol extract x 100 Weight of citral in 5ml conditioner

The fragrance delivery values for the two peaks obtained were averaged. The results are shown in table 4, with standard deviation (SD) values for each data point, and in figure 4. Table 4: % fragrance delivery results for hair swatches treated with a conditioner containing quaternised hvdrolvsed protein cvclodextrin derivative and control

% fragrance delivery for a conditioner containing 3.23 quaternised hydrolysed protein cyclodextrin (1.07)

% fragrance delivery for a conditioner without quaternised 2.06 hydrolysed protein cyclodextrin (0.64)

Figure 4: % fragrance delivery results for hair swatches treated with guaternised

hvdrolvsed protein cvclodextrin derivative and control

The study demonstrates that the conditioner containing quaternised hydrolysed protein cyclodextrin derivative delivered the fragrance, citral, to hair to a greater extent than conditioner without quaternised hydrolysed protein cyclodextrin derivative. Example 7: Measurement of the substantivity to hair of a hydrolysed protein cyclodextrin derivative according to Example 1 from shampoo and conditioner systems

Materials used in example 7 Virgin European brown hair (ex. De Meo Brothers) Basic shampoo base -10% water Basic hair conditioner -10% water Hydrolysed protein cyclodextrin derivative (as in example 1) 2% sodium lauryl ether sulphate (SLES) solution Iodine125 (Amersham Biosciences)

Preparation of test solutions A 2.0Og sample of Protein Cyclodextrin derivative was radiolabeled using the Croda Chemicals Europe standard procedure.

Croda Chemicals Europe protein radiolabelling procedure Materials: Solution 1 - Protein test solution. Solution 2 -Solution 1 adjusted to pH 7.5 and diluted 1 in 10 in sodium phosphate buffer. Solution 3 - 0.1% sodium iodide in pH 7.5 sodium phosphate buffer. Solution 4 - Sodium phosphate buffer at pH 7.5 ( 0.25M NaH2PO4^H2O [ 25.35g / 500ml ] + 0.25M Na2HPO4 [ 17.745g / 500ml ] adjusted to pH 7.5 with NaOH ). lodobeads® (Perbio Science UK Ltd) Iodine125 (Amersham Biosciences)

Procedure:

1) Rinse one iodobead with 500μl of buffer solution 4. 2) Open I125 vial and add 100μl of buffer solution 4, mix for 30 seconds using pipette. 3) Add the buffer rinsed iodobead to the vial and close the vial. 4) Leave to stand for 2 minutes. 5) Open vial and add 100μl of diluted protein solution 2, mix for 30 seconds using pipette. 6) Close vial and allow to stand for 1 hour. 7) To a 10ml glass beaker containing magnetic flea add 2ml of protein solution 1. 8) Place beaker on magnetic stirrer. 9) Open vial and remove reactant mixture by pipette and add to beaker. 10) Close vial. 11) Add 100μl of sodium iodide solution 3 to beaker. 12) Cover beaker with lead shield. 13) Turn magnetic stirrer on for 5 minutes then turn off. 14) Set up ion-exchange column with 5 ml of resin IRA-94S in chloride form. 15) Rinse column with 200ml distilled water to remove residual chloride. 16) By pipette add I125 mixture to top of column. 17) Ion-exchange at 1ml/min and collect in 10ml volumetric flask washing mix through with distilled water to the 10ml mark to generate the test solution. 18) Count the test solution, 100μl 3 times using different pipettes. 19) Store test solution in steel cabinet until ready for use.

The radiolabeled hydrolysed protein cyclodextrin derivative was added to formulations as detailed below to produce formulations containing 1 % w/w hydrolysed protein cyclodextrin derivative as supplied.

Shampoo test solution % Basic shampoo base -10% water 90.0 Radiolabeled protein cyclodextrin derivative qs (to provide 1.0% as supplied) Distilled water up to 100

Conditioner test solution % Basic hair conditioner-10% water 90.0 Radiolabeled protein cyclodextrin derivative qs (to provide 1.0% as supplied) Distilled water up to 100 Treatment of samples For each study, triplicate 100mg hair swatches were used. The level of radiolabeled protein present on the hair swatches was measured after 1 , 3 and 5 applications of the test solutions. All swatches were pre-washed in 2% SLES solution, rinsed thoroughly and dried.

The treatment protocol was as follows:

1. Each hair swatch was immersed in water (10ml) for 60 seconds at 250C. 2. Each hair swatch was dried by gently blotting between paper tissues. 3. Each hair swatch was immersed in 10ml of formulation under test for 60 seconds at 250C, ensuring good and even contact. 4. Each hair swatch was removed from the test formulation and rinsed in 3 x 250ml water at 370C; each rinse being for 30 seconds. 5. Each hair swatch was dried by gently blotting between paper tissues. 6. The radioactivity of each hair swatch was measured using a Gamma Counter.

Steps 3-5 were repeated 4 times to give five applications of each test formulation. Further radioactivity measurements were taken after 3 and 5 applications of test formulation.

Results The substantivity results, for the treatment of virgin European brown hair with the shampoo and conditioner formulations are given in tables 5 and 6 respectively and in figure 5 below. The results show that a hydrolysed protein cyclodextrin derivative is substantive to virgin European brown hair from shampoo and conditioner formulations. Table 5: Substantivitv (mg/100g hair) on triplicate virgin hair swatches after shampoo treatments 1 - 5.

Table 6: Substantivity (mg/100g hair) on triplicate virgin hair swatches after conditioner treatments 1 - 5.

Figure 5: Substantivitv of Protein Cvclodextrin from shampoo and conditioner formulations to virgin European brown hair

0 3 No. of treatments Example 8: Preparation of hydrolysed protein cyclodextrin derivative

The following steps were carried out:

1. Hydrobrazilnut AA (125g) (Croda Chemicals Europe Ltd) was loaded into a suitable beaker and stirred. 2. The pH was raised to 10.2 by the addition of 25% sodium hydroxide (19g). 3. Cavasol W7 MCT (50.25g) (available from Wacker-Chemie GmbH, Hanns- Seidel Platz 4, DE-81737 Munchen, Germany) was added over 60 minutes. 4. During addition, the pH was maintained at 10.0 - 10.2 using 25% sodium hydroxide. The reaction mixture was stirred at room temperature for a further 24 hours. The pH was maintained using 25% sodium hydroxide (11.5g). 5. 28% hydrochloric acid (20.5g) was added to lower the pH to 4.0 - 4.5. 6. Water (82.1) was added to dilute the solution. 7. Phenoxyethanol (1.4g), potassium sorbate (0.53g) and disodium EDTA (0.18g) were added and the solution stirred overnight at room temperature. The liquor was filtered through a depth filter.

Example 9: Preparation of silanised hydrolysed protein cyclodextrin derivative

The following steps were carried out:

1. Hydrotriticum WAA (125g) (Croda Chemicals Europe Ltd) was loaded into a suitable beaker and stirred. 2. The pH was raised to 10.2 by the addition of 25% sodium hydroxide (20.4g). 3. Cavasol W7 MCT (58.15g) (available from Wacker-Chemie GmbH, Hanns- Seidel Platz 4, DE-81737 Munchen, Germany) was added over 60 minutes. 4. During addition, the pH was maintained at 10.0 - 10.2 using 25% sodium hydroxide (9.Og). 5. The reaction mixture was stirred at room temperature for a further 22.5 hours. The pH was maintained using 25% sodium hydroxide (7.6g). 6. The liquor was heated to 40°C and held at this temperature. 7. The pH was adjusted to 10.0 - 10.5 using 25% sodium hydroxide (5.5g) 8. gamma-Glycidoxypropyltrimethoxysilane (Silquest A-187 silane available from GE Silicones ) (17.2g) was added dropwise via a separating funnel over a period of 95 minutes, with the pH of the reaction mixture maintained at 10.0 - 10.5 using 25% sodium hydroxide (1.Og). 9. The reaction mixture was stirred for 4 hours at 4O0C. The pH was stable in the range 10.0 - 10.5. 10. 28% hydrochloric acid (29.Og) was added to lower the pH to 4.0 - 4.5. 11. Water (10Og) was added to dilute the solution. 12. Phenoxyethanol (2.8g), potassium sorbate (Hg) and disodium EDTA (0.4g) were added and the solution stirred overnight at room temperature. 13. The liquor was filtered through a depth filter to yield a hazy filtrate.

Example 10: Smoke exposure (odour removal) studies of a protein cyclodextrin derivative according to Example 8 and Example 9 applied to hair from an aqueous solution.

Procedure.

Test material A - Hydrolysed protein cyclodextrin derivative; 5% active solution. Test material B - Silanised hydrolysed protein cyclodextrin derivative; 5% active solution. Test material C - Cavasol (beta cyclodextrin); 5% active solution Control - Water.

A hair swatch of a known weight was dipped into a solution above until thoroughly wetted. Excess liquid was squeezed out and the swatch was left to dry overnight. This was repeated for each of the above solutions The swatches were then exposed to cigarette smoke ( a cigarette was left to burn for 10 seconds in a bell jar containing the swatches and then the resultant smoke was allowed to circulate for a further 60 seconds; this procedure was repeated a further two times). Subjective sensory analysis was then carried out using a number of assessors. The swatch with the least smoke odour was ranked 1 and the swatch with the strongest smoke odour was ranked 4. 0 = no difference detected. Results.

Table 7 - Immediately after exposure.

Table 8 - 1 Hour after exposure.

Immediately after exposure there is little to distinguish between the swatches, although the hydrolysed protein cyclodextrin derivative is ranked 1st.

After 1 hour the silanised hydrolysed protein cyclodextrin derivative has significantly less smoke odour, followed by the hydrolysed protein cyclodextrin derivative. Cavasol (beta cyclodextrin) is ranked 3rd. Example 11 : Smoke exposure (odour removal) studies of a protein cyclodextrin derivative according to Example 9 applied to hair from an aqueous solution.

Procedure

Test material A - Silanised hydrolysed protein cyclodextrin derivative; 5% active solution. Test material B - Cavasol (beta cyclodextrin); 5% active solution

A hair swatch of a known weight was dipped into a solution above until thoroughly wetted. Excess liquid was squeezed out and the swatch was left to dry overnight. This was repeated for each of the above solutions The swatches were then exposed to cigarette smoke ( a cigarette was left to burn for 10 seconds in a bell jar containing the swatches and then the resultant smoke was allowed to circulate for a further 60 seconds; this procedure was repeated a further two times). Subjective sensory analysis was then carried out using a number of assessors. The swatch with the least smoke odour was ranked 1 and the swatch with the strongest smoke odour was ranked 2. 0 = no difference detected.

Results.

Table 9 - Immediately after exposure

Table 10 - 1 Hour after exposure

After immediate exposure there is no detectable difference in smoke odour but after 1 hour the swatch treated with the silanised hydrolysed protein cyclodextrin derivative was judged to have less odour compared to the Cavasol treated swatch.

Example 12: Smoke exposure (odour removal) studies of a protein cyclodextrin derivative according to Example 9 applied to hair from an aqueous solution.

Procedure

Test material A - Silanised hydrolysed protein cyclodextrin derivative; 5% active solution. Test material B - Cavasol (beta cyclodextrin); 5% active solution

A hair swatch of a known weight was dipped into a solution above until thoroughly wetted. Excess liquid was squeezed out and the swatch was left to dry overnight. This was repeated for each of the above solutions The swatches were then exposed to cigarette smoke ( a cigarette was left to burn for 10 seconds in a bell jar containing the swatches and then the resultant smoke was allowed to circulate for a further 60 seconds; this procedure was repeated a further two times). Subjective sensory analysis was then carried out using a number of assessors. The swatch with the least smoke odour was ranked 1 and the swatch with the strongest smoke odour was ranked 2. 0 = no difference detected. Results.

Table 11 - Immediately after exposure

Table 12 - 1 Hour after exposure

Immediately after exposure the silanised hydrolysed protein cyclodextrin derivative treated swatch had the least smoke odour. After 1 hour, there was little discernible difference in smoke odour. In summary, the invention comprises the following embodiments:- 1. The preparation and use of protein-cyclodextrin derivatives for use in cosmetic compositions. 2. The use of a protein cyclodextrin derivative of (1) as a fragrance retention agent in hair and skin cosmetic formulations. 3. The use of a protein cyclodextrin derivative of (1) as a malodour control agent in hair and skin cosmetic formulations. 4. The use of a protein cyclodextrin derivative of (1) as a fragrance delivery agent in hair and skin cosmetic formulations. 5. A protein cyclodextrin derivative of (1) wherein a protein and reactive cyclodextrin are reacted in the range from 50% to 200% reactive cyclodextrin by weight of active protein. The resulting derivative thus comprises 33% to 66% cyclodextrin and modified reactive amino groups of the protein in the range 10 - 100%. 6. A protein cyclodextrin derivative of (1) wherein the protein cyclodextrin derivative is the reaction product of a protein hydrolysate and a reactive cyclodextrin. 7. A protein cyclodextrin derivative of (1) wherein the protein may be derived from either animal or vegetable sources or by fermentation. 8. A protein cyclodextrin derivative of (1) wherein the protein may be in the form of a chemically modified protein (for example, quaternised or silanised) provided that some free amino groups are still present in the protein molecules. 9. A protein cyclodextrin derivative of (1) wherein the organofunctional cyclodextrin component may be any of the known cyclodextrins containing from six to twelve glucose units, especially alpha-cyclodextrin, beta- cyclodextrin or gamma-cyclodextrin cyclodextrin, more especially beta- cyclodextrin. The alpha cyclodextrin consists of six glucose units, the beta cyclodextrin consists of seven glucose units, the gamma cyclodextrin consists of eight glucose units 10. A protein cyclodextrin derivative of (1) wherein the average molecular weight of the protein component may be from 75 Daltons (D) to 500 kD, is preferably within the range from 75D to 5OkD and is more preferably in the range of from 75D to 25.5kD, even more preferably 75D to 3000D and particularly preferably 350D to 3000D expressed as weight average molecular weight (Mw) derived from size exclusion chromatography.