WO1998048811A1 - Method of fixing fragrances in fragrance composition and other compositions - Google Patents

Abstract

The present invention provides a fixative for fragrances and other isolated odorants in fragrance compositions and other personal care formulations. These fixatives prolong the presence of the fragrance on the skin. There is a proportional effect of duration related to concentration of fixatives for a given amount of fragrance components.

Description

METHOD OF FIXING FRAGRANCES IN FRAGRANCE COMPOSITION

AND OTHER COMPOSITIONS

FIELD OF THE INVENTION This invention is generally related to the fields of personal care products, cosmetics and fragrances and to compositions of matter used in consumer products. More specifically, the invention pertains to a method of fixing fragrance in fragrance compositions and personal care products containing such fragrance compositions.

BACKGROUND ART

The present invention relates to cosmetics, particularly fragrances, and to compositions of matter which contain human pheromones and which are useful in the manufacture of consumer products . Pheromones are chemicals produced by an animal or individual which elicits a specific physiological or behavioral response in another member of the same species . The human pheromones referred to in this invention include certain 16-Androstene and/or Estrene steroids, some of which occur naturally in humans.

The steroid class of Androstenes are typified by testosterone, and are characterized by a 4 -ring steroid structure with methylations at the 13 -position and at the 10- position. 16-Androstenes are further characterized by a double bond at the 16- position. Some members of this group have been reported to act as pheromones in some mammalian species-for instance, 5 alpha -Androst-16-en-3 alpha -ol and 5 alpha -Androst-16-en-3-one in pigs (Melrose, D. R. , et al . , Br . vet. J. (1971) 127:497-502). These 16-Androstenes, produced by the boar, induce mating behavior in estrus sows (Claus, et al . , Experimentia (1979) 35:1674-1675) . Some studies have noted that, in some species, various characteristics of certain 16 -Androstenes (including 5 alpha -Androst-16-en-3 alpha -ol and 5 alpha-Androst-16-en-3-one) , such as blood concentration, metabolism, and localization, are sexually dimorphic (Brooksbank, et al . , J. Endocr. (1972) 52: 239-251; Claus, et al . , J. Endocr. (1976) 68:483-484; Kwan, et al., Med. Sci. Res. (1987) 15:1443-1444). For instance, 5 alpha -Androst-16-en-3 alpha -ol and 5 alpha -Androst-16-en-3-one, as well as 4 , 16-Androstadien-3 -one, have been found at different concentrations in the peripheral blood, saliva and axillary secretions of men and of women (Kwan, T. K., et al . , Med. Sci. Res. (1987) 15:1443-1444) . The possible function of some 16-Androstenes as human pheromones, to the extent of effecting choice and judgment, has been suggested (Id.; see also Gower, et al . , "The Significance of Odorous Steroids in Axillary Odour", in, Perfumery, pgs 68-72, Van Toller and Dodd, Eds., Chapman and Hall, 1988); Kirk-Smith, D. A., et al . , Res. Comm. Psychol . Psychiat . Behav . (1978) 3:379). Androstenol (5 alpha -Androst-16-en-3 alpha -ol) has been claimed to exhibit a pheromone-like activity in a commercial men's cologne and women's perfume (Andron TM for Men and Andron TM for Women by Jovan) . Japanese Kokai No. 2295916, refers to perfume compositions containing Androstenol and/or its analogue. 5,16- Androstadien-3 beta -ol (and perhaps the 3 alpha -ol) has also been identified in human axillary secretion (Gower, et al., Supra at 57-60.

Estrene steroids are typified by 17 beta -Estradiol (1, 3 , 5 (10) -Estratrien-3 , 17 beta -diol) , and usually have a phenolic 1,3,5(10) A-ring and a hydroxy or hydroxy derivative, such as an ether or ester, at the 3- position. The human pheromones described in this application have been referred to in applicant's U.S. Ser. No. 07/707,862, filed May 31, 1991, U.S. Ser. No. 07/708,936, filed May 5, 1991, both abandoned, P.C.T. application No. PCT/US92/00219, filed Jan. 7, 1991, and P.C.T. application No. PCT/US92/00220 , filed Jan. 7, 1991. The most likely means of communication of a putative human pheromone is the inhalation of a naturally occurring pheromone present on the skin of another. Several 16-Androstene steroids, including 5 alpha -Androst-16-en-3 alpha -ol and 5 alpha-Androst- 16-en-3-one, 4 , 16-Androstadien-3-one and 5 , 16-Androstadien-3 beta-ol, are naturally occurring in humans and may be present on the skin. It is estimated that the naturally occurring maximum concentration of all 16-Androstene steroids on human skin is from 2 to 7 ng/cm<2>. The subject invention is effective because it delivers a much larger amount of the active pheromone steroids to the skin than is normally present. There is however, little agreement in the literature as to whether or not any putative pheromone actually plays a role in the sexual or reproductive behavior of mammals, particularly of humans. See: Beauchamp, G. K. , et al . , "The Pheromone Concept in Mammalian Chemical Communication: A Critique", in: Mammalian Olfaction, Reproductive Processes, and Behavior, Doty, R. L., Ed., Academic Press, 1976. See also, Gower, et al . , supra at 68-73.

Receptors for pheromones are found in the vomeronasal organ (VNO) , a small structure which opens to the nasal passage in normal individuals (Moran, D. T., et al., J. Steroid Biochem. and Molec . Biol . (1991) 39:545; Stensaas, L. J. , et al . , J. Steroid Biochem. and Molec. Biol. (1991) 39:553; Garcia-Velasco, et al . , J. Steroid Biochem. and Molec. Biol. (1991) 39:561). An odor does not bind to a VNO receptor-only a pheromone. A pheromone-specific change in the electrical potential of VNO receptor epithelium can be measured as described by Monti-Bloch, L., et al . (J. Steroid Biochem. and Molec. Biol. (1991) 39:573). This receptor binding activity is an essential characteristic of an active pheromone.

The compositions of many commercial perfumes and fragrances contain natural mammalian pheromones. Since pheromones are generally species specific, the natural mammalian pheromones found in commercial perfumes do not function as a pheromone, but instead provide a fixative note in the overall composition of the fragrance. Thus these perfumes, personal care products and cosmetics do not bind to pheromone receptors in the VNO and do not stimulate the vomeronasal nerve which communicates with the hypothalamus of the brain. Furthermore, in some cases the use of natural animal pheromones, or synthetics related to animal pheromones, may cause skin irritations or allergic responses in some individuals. Still further, since the source of animal pheromones used in fragrances are the anal glands of the contributing animal some individuals find it objectionable to use these substances .

SUMMARY OF THE INVENTION The invention provides a method for fixing fragrance in non- herapeutic, fragrance compositions containing a perfumery odorant, by adding a human pheromone which serves as a fixative.

The fixatives include steroidal compounds having human pheromone activity such as Androsta-4, 16-dien-3-one, Androsta-4 , 16-dien-3 alpha -ol, Androsta-4 , 16-dien-3 beta -ol, 19-nor-4,16- Androstadien-3-one, 19-nor-10-OH-4 , 16-Androstadien -3one, 19-OH- , 16-Androstadien-3-one 1,3,5(10) ,16- Estratetraen-3-ol methyl ether, 1, 3 , 5 (10) , 16-Estratetraen -3-yl acetate, 1, 3 , 5 (10) , 16-Estratetraen-3-yl propionate, 1, 3 , 5 (10) , 16-Estratetraen-3-ol, and any combinations thereof .

DESCRIPTION OF THE DRAWING FIG. 1 schematically illustrates the synthesis of 10 -hydroxy-4 , 16-estradien-3-one .

FIG. 2 schematically illustrates the synthesis of Androsta- , 16-dien-3-one, Androsta-4 , 16-dien-3 alpha -ol, and Androsta-4 , 16-dien-3 beta -ol. FIG. 3 schematically illustrates the synthesis of 19-nor 4 , 16-Androstadien-3-one .

FIG. 4 schematically illustrates syntheses of 19-OH-Androst- 4 , 16-dien-3-one .

FIG. 5 schematically illustrates an alternate synthesis of 19-OH-Androsta-4 , 16-dien-3 -one .

FIG. 6 schematically illustrates synthesis of 1, 3, 5 (10) , 16-Estratetraen-3-ol.

FIG. 7 schematically illustrates an alternate synthesis of Androsta- , 16-dien-3-one .

DETAILED DESCRIPTION

Before the present compositions are disclosed and described, it is to be understood that this invention is not limited to specific fragrances, specific steroidal compounds, or the like, as such components may, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended as limiting.

It must be noted that, as used in the specification and the appended claims, the singular form "a", "an" and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "a perfumery odorant" includes mixtures of perfumery odorants, reference to "a human pheromone" includes mixtures of human pheromones, and the like.

A. Definitions An "environmental fragrance" is a fragrance or odor which is used to odorize a volume of air rather than an individual or object. The source of the environmental fragrance may be an object, for example an object composed to gradually release a fragrance into the adjacent air.

An "odor" is any scent or smell, whether pleasant or offensive. An odor is consciously perceived by an individual when odorant molecules bind to the olfactory epithelium of the nasal passage. A "perfumery odorant" is an odorant used for the principal purpose of providing a odor. A "scent" is the odor left behind by an animal or individual. People use perfumes to augment their natural scent .

A "perfume" or a "fragrance composition" is a specific pleasantly odorous cosmetic composition for topical application to an individual . Technically, perfumes are mixtures of a variety of substances, and may include natural materials of vegetable or animal origin, wholly or partly artificial compounds, or mixtures thereof. Dissolved in alcohol, these mixtures of various volatile fragrant substances release their scents into the air at normal temperatures. The extrait-the mixture which contains the highest proportion of fragrance concentrate and the least possible alcohol-is called perfume. Mixtures of lower concentration include eau de parfum, after shave, eau de toilette, eau de sport, splash cologne, eau de cologne, cologne, eau fraiche, and the like. In addition to the fragrance solutions which are diluted with alcohol, there are also those which are diluted with oil. Furthermore, compact and cream perfumes are produced by mixing up to 25% fragrance oil with solids such as paraffin or other waxes. Generally all the fragrance compositions described above are referred to as perfumes, and that is how the term is used herein. A "pheromone" is a chemical secretion which elicits a specific physiological or behavioral response in another member of the same species. In addition to physiological responses, pheromones can be identified by their species specific binding to receptors in the vomeronasal organ (VNO) . Thus, human pheromones bind to human receptors. This can be demonstrated by measuring the change in the summated potential of neuroepithelial tissue in the presence of the pheromone. Human pheromones induce a change of at least about - 5 millivolts in human neuroepithelial tissue of the appropriate sex (The binding of pheromones is generally gender specific) . Naturally occurring human pheromones induce gender specific changes in receptor binding potential in vivo in the human VNO. Naturally occurring human pheromones can be extracted and purified from human skin and they can also be synthesized, as described herein. "Human pheromones" are pheromones which are effective as a specifically binding ligand in human VNO tissue, regardless of how the pheromone was obtained. Thus, both a synthesized and purified substance may be considered a human pheromone.

"Gender specific" refers to a difference in the effect of, or response to, a compound or composition between males and females of the same species. "Tissue paper" is a soft, fibrous, absorbent paper such as the type commonly used as a disposable handkerchief or as toilet paper. The "vomeronasal organ" is a cul-de-sac which opens to the nasal passage in humans and contains specialized receptor cells for pheromones. A "fixative" or fixing agent is an agent which prolongs the odor of a fragrance when placed on a substrate, such as skin, paper and the like.

B. Perfumes The art and science of perfumery has been developed over several hundred years and is now well established. A brief summary of perfumery is provided herein. This subject is treated more fully in many publications including Wells, F. V. and M. Billot, Perfumery Technology, Ellis Horwood, Ltd. , publisher, 2nd Ed. 1981.

1. Types of Ingredients

The diversity of the non-animal, natural products used in perfumery is considerable. In addition, advances in organic chemistry in the later nineteenth and the twentieth centuries have provided an equally broad diversity of artificial odorants as well as the ability to synthesize some of the naturally occurring components of natural odorants. Most perfumes combine preparations of naturally occurring materials with synthetic odorants. The natural odorants that are generally employed in perfumery come from both animal and vegetable materials and can be assigned to the following six categories based on how they are treated:

1) Concrete oils-extracted with hydrocarbon solvents, without heat;

2) Absolute oils-alcohol extracted from concrete oils, without heat;

3) Essential oils-distilled from naturally occurring materials; 4) Expressed oils-physically removed directly from the natural material;

5) Isolates-fractionally distilled from essential oils; 6) Tinctures-obtained by prolonged alcohol extraction of naturally occurring materials.

A perfumer will typically have numerous oils, isolates, and tinctures from a variety of natural sources within each category. The perfumer will also have a vast array of artificial odorants and synthetics of naturally occurring compounds. The art of perfumery involves the mixing of these various materials to produce a finished fragrance . While there are many subjective approaches to the formulation of a perfume, most seem to incorporate the notion of top notes, middle notes and base notes. Top notes are very volatile and lack tenacity, or staying power. Middle notes are somewhat lower in volatility and are used as modifiers of the top notes. Base notes are still lower in volatility and are long-lasting in odorous effect. Base notes are also referred to as fixatives of the fragrance. Notes of animal origin, or artificials which mimic animal notes, are usually base notes.

2. Animal Notes

Many commercial perfumes contain notes from animal sources, usually pheromones of the species from which the material is obtained, or synthetics and artificial notes which mimic the characteristics of animal notes. The principal animal -derived notes are the following:

1) musk-derived from the scent gland of the musk deer;

2) civet-obtained as a glandular secretion of the civet cat; 3) castoreum-obtained from the preputial follicle of the beaver; and

4) ambergris-a regurgitated or excreted material obtained from sperm whales. The first three are pheromones for the species of origin, but since pheromones are species specific, they do not induce any pheromone-related behavior in humans. Animal notes are used as a fixative for the perfume fragrance . As a concentrate the odor of animal notes may not be pleasing, but when diluted, they contribute to the fragrance of the final product.

3. Human Pheromones

In the subject invention, human pheromones or analogs thereof, are used instead of, or in addition to animal pheromones, or their derivatives or homologues . As a component in compositions of matter human pheromones have several advantages .

Perfumed products that do not contain human pheromones do not stimulate VNO receptors since an odorant which is not a pheromone for humans stimulates only the olfactory receptors of the nose. Fragrance compositions which are both pleasant smelling and also contain human pheromones will stimulate both olfactory receptors, and pheromone receptors in the VNO of individuals. Such a fragrance composition provides a broader olfactory stimulation than previously possible.

Some ingredients associated with commonly used animal notes (e.g. benzyl benzoate, paracresol, nitro-musks) have been found to cause skin irritation in some individuals. Furthermore, some individuals report an allergic response to some perfumes. Fragrance compositions containing naturally occurring human pheromones would be less likely than commonly used animal-related components to cause irritation or allergic response .

Finally, a perfume which uses human pheromones rather than material derived from the anal or preputial glands of animals would be inherently more appealing to many consumers .

As a concentrate, pheromones may or may not have a detectable odor. Since they bind to receptors which are physically and functionally distinct from olfactory receptors, they may or may not carry their own smell. However, some of the pheromones described herein do in fact have an odor. As a concentrate, the odor of these pheromones may not necessarily be pleasant. Thus, when diluted in a perfume the practical upper concentration limit is determined by the pleasantness of the resulting fragrance. Generally, human pheromones are present in the fragrance composition as a fixative of the subject invention at a concentration of no more than about 200 mu g/ml, more commonly no more than about 100 mu g/ml, preferably no more than about 50 mu g/ml, and more preferably no more than about 25 mu g/ml.

Generally, human pheromones are present in the fragrance composition of the subject invention at a concentration of at least about 50 ng/ml , but it will depend upon the concentration of fragrant substances present. There is a direct effect on prolonged smell related to the pheromone concentration, because the more fixative used, in general, the longer the fragrance persists .

C. Other Products Containing Pheromones

Perfumes are commonly used per se as a personal care product. However, odours can be used in a variety of personal care products, household products and industrial products. The use of human pheromones per se, or perfumes containing human pheromones in these other products falls within the scope of the subject application. 1. Personal Care Products

Fragrances containing human pheromones can be used in the preparation of cosmetics, make-up preparations, toilet and beauty preparations, bath and beauty soaps, bath oils, face and body creams and oils, underarm deodorants and the like. The preparations of these personal care products are known to those skilled in the art. These products frequently contain a fragrance. A human pheromone or a fragrance containing human pheromones is added to these products in the same way that fragrance per se may be added.

2. Environmental Odorants

Pheromones or fragrances containing human pheromones may also be used as environmental odorants as in air fresheners and the like. The fragrance and pheromone can be dispensed into the air by use of an aerosol dispenser, or by preparations of liquid, gel or solid compositions containing fragrance and pheromone which slowly release the pheromone, or fragrance and pheromone, into the air by exposure of the composition to the atmosphere .

For aerosol administration, the active ingredient is preferably supplied in a liquid or finely divided form along with a surfactant and a propellant. Typical percentages of active ingredients are 0.001 to 2% by weight, preferably 0.004 to 0.10%.

Surfactants must, of course, be nontoxic, and preferably soluble in the propellant. Representative of such agents are the esters or partial esters of fatty acids containing from 6 to 22 carbon atoms, such as caproic, octanoic, lauric, palmitic, stearic, linoleic, olestearic and oleic acids with an aliphatic polyhydric alcohol or its cyclic anhydride such as, for example, ethylene glycol, glycerol, erythritol, arabitol, mannitol, sorbitol, and hexitol anhydrides derived from sorbitol (the sorbitan esters sold under the trademark "Spans") and the polyoxyethylene and polyoxypropylene derivatives of these esters. Mixed esters, such as mixed or natural glycerides, may be employed. The preferred surface-active agents are the oleates or sorbitan, e.g., those sold under the trademarks "Arlacel C" (sorbitan sesquioleate) , "Span 80" (sorbitan monoleate) and "Span 85" (sorbitan trioleate) . The surfactant may constitute 0.1-20% by weight of the composition, preferably 0.25-5%. The balance of the composition is ordinarily propellant. Liquefied propellants are typically gases at ambient conditions, and are condensed under pressure. Among suitable liquefied propellants are the lower alkanes containing up to five carbons, such as butane and propane; fluorinated or fluorochlorinated alkanes, such as are sold under the trademark "Freon" . Mixtures of the above may also be employed.

In producing the aerosol, a container equipped with a suitable valve is filled with the appropriate propellant, containing the finely divided active ingredient and surfactant . The ingredients are thus maintained at an elevated pressure until released by action of the valve. An alternative means of releasing fragrance and pheromone into a designated air space is by means of gradual evaporation and release into the atmosphere from a liquid, semi-solid or solid composition containing or a fragrance and pheromone. A fragrance containing a human pheromone may be incorporated into the composition in a variety of ways depending on the nature of the composition.

If the composition is a liquid, gel, cream or ointment, and the pheromone ingredient is soluble in the composition it can simply be dissolved in the composition. If the pheromone ingredient is slightly soluble or insoluble in the composition, a suspension can be prepared by addition and mixing. In some cases such as room odorants, car odorants and the like, the composition containing pheromone is applied in a liquid state and remains liquid during evaporation. In other cases, such as paints and the like, the composition containing pheromone is applied as a liquid and then solidifies, leaving the pheromone to slowly evaporate from the solid.

If the composition is solid, the pheromone ingredient can be added by first melting the solid up to a maximum temperature of 100 degrees C, preferably 75 degrees C, more preferably 50 degrees C, adding the pheromone ingredient and then allowing the mixture to cool and solidify. This approach may be used with wax or resin for example. Alternatively, the pheromone ingredient may first be mixed in a volatile solvent such as ethanol, dimethyl sulfoxide or the like, and then mixed with an absorbent solid composition such as tissue paper, cloth and the like. The solvent then evaporates leaving the pheromone residue in the solid composition, from which the pheromone slowly evaporates into the atmosphere . The uses of fragrance compositions containing human pheromones, as provided herein, are examples of alternative uses which fall within the intended scope of the claims and do not limit the intended scope of use of this invention.

3. Other Products

This mixture includes other products such as fibrous materials which will absorb pheromones with fragrances. For instance, cloth, papers (including tissue papers) , clothing, paper towels, stationery and the like.

D. Human Pheromones

As described herein human pheromones generate a change in receptor potential in the VNO of human subjects .

16-Androstene steroids are aliphatic polycyclic hydrocarbons characterized by a four-ring steroidal structure with a methylation at the 13- position, and a double bond between the 16- and 17- positions. An

Androstene steroid is commonly understood to mean that the compound has at least two methylations, at the 13-position and the 10- position, thereby creating 18- position and 19- position carbons respectively. Unless a compound is explicitly described as "19-nor" it is understood that the compound does have a 19- carbon group. However; it is intended that 19-nor-16-Androstenes are generally regarded as 16 -Androstene steroids for the purpose of the present invention. Estrene steroids are aliphatic polycyclic hydrocarbons with a four-ring steroidal structure, usually an aromatic 1,3,5(10) A-ring, a methylation at the 13 -position and a hydroxyl at the 3 -position. In describing the location of groups and substituents of 16-Androstene and Estrene steroids, the following numbering system will be employed.

1. 16 -Androstenes Useful in Conjunction with the Invention

The invention is directed to fragrance compositions containing a human pheromone which may be included in a group known as Androstene steroids of which testosterone (17-hydroxy-4-androstene-3-one) is an example, and to combinations of Androstene and Estrene steroids. Specifically included are those steroids disclosed in the U.S. patent application Ser. No. 07/903,604 filed Jun. 24, 1992, the entirety of which is incorporated by notice. 16 -Androstenes are further characterized by a double bond at position 16-.

The 16 -Androstenes of this invention have the formula : I

wherein Rl is selected from the group consisting of oxo, alpha -( beta -)hydroxy, alpha -( beta -) acetoxy, alpha -( beta -) propionoxy, alpha -( beta -)methoxy, alpha -( beta -) lower acyloxy, alpha -( beta -) lower alkyloxy, and alpha -( beta -) benzoyloxy; R2 is selected from the group consisting of hydrogen, hydroxy, acyl, acyloxy, alkoxy, methyl, hydroxymethyl , acylmethyl, acyloxymethyl, alkoxymethyl, lower alkyl, hydroxyalkyl, acylalkyl, acyloxyalkyl, and alkoxylalkyl ; and "a" and "b" are alternative sites for an optional double bond. Preferred embodiments include 4,16 Androstadien-3-one (Rl = oxo, a = double bond, R2 = methyl, commercially available from Steraloids, Inc., also referred to as Androstadienone) , and 19-hydroxy-4 , 16-androstadien-3-one (Rl = oxo, a = double bond, R2 = hydroxymethyl), 4 , 16-Androstadien-3 alpha ( beta ) -ol (Rl = hydroxy, a = double bond, R2 = methyl), 19-nor-4, 16-Androstadien-3-one (Rl= oxo, a = double bond, R2 = hydrogen), and 19-nor-10-OH-4 , 16 Androstadien-3-one (Rl = oxo, a = double bond, R2 = hydroxy) , syntheses of which are described herein) .

2. Estrenes Useful in Conjunction with the Invention The invention is additionally directed to fragrance compositions containing a human pheromone which may be included in a group of Estrene Steroids, or to combinations of Estrene and 16 -Androstene steroids.

Specifically included are those steroids disclosed in the

U.S. patent application Ser. No. 07/903,604, filed Jun.

24, 1993, the entirety of which is incorporated by notice. These Estrenes are structurally similar to

Estradiol (also referred to as 1 , 3 , 5 (10) -Estratriene-3 , 17 beta -diol) , but are distinguished from Estradiol by the double bond at the 16 -position.

These Estrenes have the formula: II

wherein R4 is selected from the group consisting of hydrogen, and alkyl; R6 is selected from the group consisting of hydrogen, lower alkyl, lower acyl, benzoyl, cypionyl, acetyl, glucuronide, propionyl, and sulfate; and "c" is an optional double bond.

These Estrenes can be distinguished from each other by variations at the 3 -position, variations at the 17-position and variations at the 16-position, with an optional double bond at the 16 -position. Preferred embodiments include 1, 3 , 5 (10) , 16-Estratetraen-3-ol, available from Research Plus, Inc. and from Steraloids, Inc .

E. Synthesizing Human Pheromones As indicated in Section Dl, above, some of the preferred 16 -Androstene pheromones are not commercially available. Their syntheses are provided herein.

1. Synthetic Methods a. Preparation of 3-Position, 5-Position, and 19-Nor

Derivatives As shown in formula I, above, the compounds used in the methods of the present invention are 16 -Androstene steroids substituted at the 3-, 5-, and 19-positions . Many of the 3- and 5- substituted steroids are known compounds which may be derived from 17-hydroxy-and 17-oxosteroids (commercially available e.g. from Aldrich Chemical Co) by elimination or reduction to the DELTA 16 homologue. The syntheses of most of these compounds are described by Ohloff (supra) .

Alkoxy derivatives are prepared from their corresponding hydroxy steroids by reaction with an alkylating agent such as trimethyloxonium fluoroborate, triethyloxonium fluoroborate or methylfluorosulfonate in an inert chlorocarbon solvent such as methylene chloride . Alternatively, alkylating agents such as alkyl halides, alkyl tosylates, alkyl mesylates and dialkylsulfate may be used with a base such as silver oxide or barium oxide in polar, aprotic solvents as for example, DMF, DMSO and hexamethylphosphoramide .

General procedures, for synthetic reactions of steroids are known to those skilled in art (See for example, Fieser, L. F. and M. Fieser, Steroids, Reinhold, N.Y. 1959) . Where time and temperature of reactions must be determined, these can be determined by a routine methodology. After addition of the required reagents, the mixture is stirred under an inert atmosphere and aliquots are removed at hourly intervals. The aliquots are analyzed by means of thin-layer chromatography to check for the disappearance of starting material, at which point the work-up procedure is initiated. If the starting material is not consumed within twenty-four hours, the mixture is heated to reflux and hourly aliquots are analyzed, as before, until no starting material remains. In this case the mixture is allowed to cool before the work-up procedure is initiated.

Purification of the products is accomplished by means of chromatography and/or crystallization, as known to those skilled in the art.

1. Synthesis of 10-Hydroxy-4 , 16-Estradien-3-one

As depicted in FIG. 1, starting from 5 (10), 16- estradien-3 -one . b. Preparation of 19-OH Derivatives 1. Synthesis of 19-OH-4,16 -Androstadien-3-one This compound has been disclosed as an intermediate in the synthesis of 19-oxo-3-aza-A-homo-5 beta -androstane (Habermehl, et al . , Z. Naturforsch. (1970) 25b:191-195) . A method of synthesizing this compound is provided. Additional methods of synthesis are provided in Example 11.

EXAMPLES The following examples are provided for illustrative purposes and should not be construed as limitations of the invention described in this application.

Abbreviations used in the examples are as follows aq. = aqueous; RT. = room temperature; PE = petroleum ether (b.p. 50o-70o) ; DMF = N, N-dimethylformamide; DMSO = dimethyl sulfoxide; THF = tetrahydrofuran.

EXAMPLE 1 Estra-4, 16-dien-10jβ-ol-3-one, 4 To a solution of estra-5(10), 16-dien-3-one (256.4 mg, 1.000 mmol) in 6 ml of 1, 2-dimethoxyethane (DME) was added MCPBA (189.8 MG, 1.100 mmol) in 6 mL of DME + 2.4 mL of water. After stirring ^XA h, the reaction mixture was poured into 30 g of 5% (w/w) sodium thiosulfate pentahydrate and extracted three times with 30 mL aliquots of ethyl acetate. The combined organic extracts were washed with 30 L of saturated sodium bicarbonate + three 30 mL portions of brine, dried over magnesium sulfate, and filtered through diatomaceous earth. The residue was washed with 10 mL of ethyl acetate and the combined filtrates were concentrated under reduced pressure. To the resulting crystalline film was added 45 mL of 5% (w/v) potassium hydroxide in methanol and the mixture was refluxed with exclusion of moisture for 1 h, after which it was poured into 100 mL of ice water and extracted three times with 70 mL aliquots of ether. The combined ethereal extracts were washed three times with 70 mL portions of brine, dried over magnesium sulfate, and filtered through diatomaceous earth. The residue was washed with 25 mL of ether and the combined filtrates were concentrated under reduced pressure. Preparative TLC (50% ethyl acetate/hexanes on alumina GF, 1000 μ) of the residual resin, followed by recystallization from aqueous ethanol gave light yellow needles (62.4 mg, 0.229 mmol, 23%), m.p. 156-166°C. TLC (50% ethyl acetate/hexanes on silica gel; estrone R_f 0.59) showed a major product (R_f 0.44) with minor contaminants at R_f 0.62 and 0.73.

EXAMPLE 2

Androsta-4 , 16-dien-3-one (4) This synthesis is depicted in FIG. 2. Several methods are known for the conversion of testosterone into Androsta-4 , 16-dien-3-one (Brooksbank et al . , Biochem. J. (1950) 47:36). Alternatively, thermolysis (460o) of the methyl carbonate of testosterone gives Androsta-4 , 16- dien-3-one in 90% yield. 17 beta -Methoxycarbonyloxy- androst-4-en-3-one (IV) was prepared from testosterone (III. Fluka) with methyl chloroformate/pyridine (a) in 76% yield (after recrystallization from MeOH) . M.p. 140o-141o, [a] D = + 95.4o (C = 1.10) - IR. (CDC13) : 1740s, 1665s, 1450s, 1280s, <1> H-NMR. (360 MHz): 0.87 (s, 3 H) ; 1.20 (s, 3 H) ; 3.77 (s, 3 H) ; 4.53 (br. t, J = 8, 1 H) ; 5.75 (s, 1 H) . A solution of the methyl carbonate IV in toluene was pyrolyzed (b) as described for 1. Recrystallization of the crude product from acetone at RT. gave pure ketone 4 in 90% yield. M.p. 127o-129.5o, [a] D = + 118.9o (c = 1.32) ([3]: m.p. 131.5o-133.5o (hexane) , [a] D <16> + 123o +/- 3.5o (c = 1.03)). - IR. (CDC13) : 3050w, 1660s, 1615m. - <1> H-NMR. (360 MHz) : 0.82 (s, 3 H) ; 1.22 (s, 3 H) ; 5.70 (m, 1 H) ; 5.73 (S, 1 H) ; 5.84 (m, 1 H) . EXAMPLE 3 Androsta-4 , 16-dien-3 alpha -ol (5) and -3 beta -ol (6) These syntheses are depicted in FIG. 2. Androsta-4 , 16-dien-3-one (4) was reduced at - 55o with lithium tris (1, 2-dimethylpropyl) hydridoborate in THF (c) as described for the preparation of 2 (FIG. 1) . Chromatography on silica gel with CH2C12/ethyl acetate 9:1 gave pure axial alcohol 15 (48%, yield) and pure equatorial alcohol 6 (48% yield) . Analytical samples were further purified by recrystallization (from PE at - 30o for 5, from cyclohexane at RT. for 6) .

Data of 5. M.p. 77o-79o, [a] D = + 120.6o (c = 1.26) - IR. (CDC13) : 3620m, 3440m br . , 1660m, 1595w. - <1> H-NMR. (360 MHZ): 0.79 (s, 3 H) ; 1.02 (s, 3 H) ; 4.07 (m, w % approximately equal 10, 1 H) ; 5.48 (d x d, J = 5 and 2, 1 H) ; 5.71 (m, 1 H) ; 5.85 (m, 1 H) .

Data of 6. M.p. 116o-119o, [a] D = + 53.9o (c = 1.28) ([47] : m.p. II60-II80, [a] D = + 59.3o (c = 0.4) - IR. (CDC13) : 3610m, 3420m br . , 3050m, 1660m, 1590w. - <1> H-NMR. (360 MHz): 0.78 (s, 3 H) ; 1.08 (s, 3 H) ; 4.15 (m, w approximately equal 20, 1 H) ; 5.30 (m, w approximately equal 5, 1 H) ; 5.71 (m, 1 H) ; 5.85 (m, 1 H) .

EXAMPLE 4 19-nor-Androsta-4, 16-dien-3-one (9)

This synthesis is depicted in FIG. 3. 19-Nortestosterone (XIX) is commercially available, e.g. from Chemical Dynamics Corp. It provides the starting material for 19-Nor-16-androsten derivatives. 19-Nor- testosterone (XIX) (Chemical Dynamics Corp.) was converted into the known acetate (Hartman, J. A. et al . , J. Am. Chem. Soc. (1956) 78:5662) with acetanhydride and pyridine (a) . A solution of this acetate (4.8 g, 15.17 mmol) in toluene (10 ml) was pyrolyzed (b) at 540o (200 Torr, slow N2 -stream) in a glass tube packed with quartz pieces. Chromatography of the crude pyrolysate (3.1 g) on silica gel (150 g) with CH2C12 gave 1.1 g (28%) of the homogenous oily ketone 9; [a] D = + 57.9o (c = 1) ([27] : m.p. 71o-73o) . - IR. (CHC13) : 1660s, 1615m, 1585w, - <1> H-NMR. (90 MHz): 0.84 (s, 3 H) ; 5.82 (m,- 2 H) ; 5.87 (br. s, 1 H) .

EXAMPLE 5 Syntheses of 19-OH-4 , 16-Androstadien-3 -one (19) - 6β , 19-Epoxy-17-iodoandrosta-4 , 16-diene 3-ethylene ketal, 18:

Refer to Figure 4. A mixture of crude 6/3, 19- epoxy-53-chloro-17-iodoandrost-16-en-3 -one (37,1.38 g, 3.09 mmol) (G. Habermehl and A. Haaf, Z. Naturforsch . 1970,. 25b, 191-195), ethylene glycol (0.97 g, 16 mmol), toluene (50 mL) , and p-toluenesulfonic acid monohydrate (20.3 mg, 0.107 mmol) was refluxed 19 h with azeotropic removal of water (Deen-Stark) . After cooling ethyl acetate (100 mL) was added and the reaction mixture was washed with 100 mL of saturated sodium bicarbonate + 100 mL of brine. The organic phase was dried over magnesium sulfate and filtered through diatomaceous earth. The residue was washed with 25 mL of ethyl acetate and the combined filtrates were concentrated under reduced pressure to give a tan, crystalline solid (1.47 g) . This residue was suspended in anh. methanol (40 mL) , potassium acetate (2.44 g, 24.9 mmol) was added, and ca. 26 mL of methanol were distilled off. The remainder was concentrated under reduced pressure, water (50 mL) was added, and the mixture was extracted three times with 25 mL aliquots of methylene chloride. The dried (sodium sulfate) extracts were filtered through diatomaceous earth and the residue was washed with 10 mL of methylene chloride. Concentration of the combined filtrates under reduced pressure gave a yellow solid, which was purified further by flash chromatography (5-7.5-10% ethyl acetate/methylene chloride on silica gel) and recrystallization from methanol to give light yellow needles (914.6 mg, 2.013 mmol, 65%), m.p. 187-189°C. ¹H- NMR: 6.13 3, 1H, dd, 16-H; 5.82 3, 1H, s, 4-H; 4.71 3, 1H, d, 6α-H; 4.22 3 and 3.53 3, 2H, AB, 19-H's; 4.10-3.28 3, 4M, mult., 3-ketal H's; 0.83 3, 3H, s, 18-Me.

Androsta-4 , 16-dien-19-ol-3-one , 19 : Refer to Figure 5. Anh. Amonia (ca. 75 mL) was distilled through a KOH tower into a 250 mL flame-dried 3 -neck flask fitted with an inlet adapter, a magnetic stirring bar, a dry ice/acetone condenser, and a stopper. A solution of 6β , 19-epoxy-17-iodoandrosta-4 , 16-diene 3- ethylene ketal (18_., 880.4 mg, 1.938 mmol) in dry tetrahydrofuran (THF, 45 mL) was added, followed by metallic sodium (0.20 g, 8.7 mg-atom) cut in small pieces. After stirring under argon pressure for 30 min. the reaction was quenched with the addition of abs . ethanol (1.0 mL) . Ammonia was allowed to boil off overnight, 50 mL of water were added, and the mixture was extracted with three 25 mL portions of methylene chloride. The combined organic extracts were washed with 50 mL of brine, dried over magnesium sulfate, and filtered through diatomaceous earth. After washing the residue with 10 mL of methylene chloride the combined filtrates were concentrated under reduced pressure. The intermediate ketal proved remarkably unreactive, but was finally hydrolyzed by refluxing 18 h in 5 L of chloroform and 2.5 mL of 4 N hydrochloric acid. To the cooled hydrolysis mixture ethyl acetate (50 mL) was added and the layers were separated. The organic phase was washed with 25 mL of saturated sodium bicarbonate + 25 mL of brine, dried over magnesium sulfate, and filtered through diatomaceous earth. The residue was washed with 10 mL of ethyl acetate and the combined filtrates were concentrated under reduced pressure. The resulting brown foam was purified by flash chromatography (50% ethyl acetate/hexanes on silica gel) followed by preparative TLC (50% ethyl acetate/hexanes on silica gel GF, lOOOμ thickness) to give a partially crystalline film (66.7 mg, 0.233 mmol, 12%). ^-NMR: 5.92 3, 1H, s, 4-H; 5.87-5.64 3, 2H, mult., 16,17-H's; 4.10 3, and 3.94 3, 2H, AB, 19- H's; 0.79 3, 3H, s, 18-Me.

EXAMPLE 6 Synthesis of 19-OH-5 , 16-Androstadien-3-j6-ol The following method of synthesis is depicted in FIG. 5: 3 , 19-Dihydroxyandrost-5-en-17-one tosylhydrazone (20)

A suspension of androst-5-en-33, 19-diol-17-one { 1 , commercially available from Research Plus, 512.5 mg, 1.684 mmol) and p-toluenesulfonylhydrazide (p-TsNHNH₂, 392.1 mg, 2.105 mmol) in 2-propanol (6.0 mL) was refluxed 24 h. To the cool reaction mixture were added 20 mL of ether and the solvent was removed under reduced pressure. The residue was taken up in 10 mL of ether and the solution was filtered through diatomaceous earth. 10 mL of hexanes were added to the filtrate and the suspension was concentrated under reduced pressure. Residue was taken up in 10 L of hot benzene and the cooled suspension was filtered. The filtrate was concentrated under reduced pressure and then flash chromatographed (40% ethyl acetate/hexanes on silica gel) to give an opaque resin (0.69 g, 1.6 mmol, 87%).

Androsta-5 , 16-dien-3ff, 19-diol, 3: Refer to Figure 6. A solution of androst-5-en-3(β, 19-diol-17- (p- toluenesulfonyl) hydrazone {2 , 0.69 g, 1.6 mmol) in anh. Tetrahydrofuran (THF, 35 mL) was cooled in an ice/acetone bath under argon and n-butyllithium (2.6 M in hexanes, 3.7 mL, 9.3 mmol) was added dropwise, with stirring, over the period of 1 min. The reaction mixture was stirred 4 days, during which time it was allowed to gradually warm to room temperature. The reaction was then poured into 50 mL of ice-saturated ammonium chloride and the layers were separated. The aqueous layer was extracted twice with 25 mL portions of ethyl acetate. The combined organic phases were washed with 25 mL of saturated sodium bicarbonate + 25 L of brine, dried over magnesium sulfate, and filtered through diatomaceous earth. The residue was washed with 10 mL of ethyl acetate and the combined filtrates were concentrated under reduced pressure. The residual yellow resin was flash chromatographed (50-55-60% ethyl acetate/hexanes on silica gel) and crystallized from methyl t-butyl ether/benzene to give fluffy white crystals (92.5 mg, 0.361 mmol, 24%), m.p. 169-171°C.

EXAMPLE 7 Alternate synthesis of Androsta-4 , 16-dien-3-one (25) The following method of synthesis is depicted in FIG. 7: Dehydroepiandrosterone p-Toluenesulfonylhydrazone (23)

Dehydroepiandrosterone (VII) (14.4 g, 50.0 m mole) and p-toluenesulfonylhydrazide (12.75 g, 68.5 m mole) in dry methanol (300 ml) were heated under reflux for 20 hours. The mixture was transferred to a conical flask and allowed to cool. The crystalline product was filtered off under suction and washed with methanol (50 ml) . Further crops of product were obtained by sequentially evaporating the filtrate to 75 ml and 20 ml, and allowing to crystallize each time. Total yield was 21.6 g (95%) .

Androsta-5,16-dien-3;β-ol (24) Dehydroepiandrosterone p-toluenesulfonylhydrazone

(23) (22.8 g, 50.0 m mole) in dry tetrahydrofuran (1.0 liters) was cooled in a dry ice/isopropanol bath. The mixture was stirred while n-butyl lithium (125 ml of 1.6M solution in hexane, 200 m mole) was added. The mixture was allowed to warm to room temperature and was stirred for 24 hours. Water (50 ml) was added with cooling in ice. The mixture was poured into saturated ammonium chloride solution/ice (500 ml) and extracted with ether (x2) . The organic layers were washed with saturated sodium bicarbonate solution (500 ml) and saturated sodium chloride solution (500 ml) , dried (MgS04) and evaporated in vacuo to give the crude product. This was purified by flash chromatography on 190 g silica gel 60, 230-400 mesh, eluting with ethyl acetate/hexane (20:80 arrow right 50:50) to give crystalline material. The product was recrystallized from methanol (45 ml) /3% hydrogen peroxide (8 ml) washing with methanol (30 ml) /water (8 ml) to give pure product (6.75 g, 50%) .

Androsta-4 , 16-dien-3-one (25)

A solution of 10 g of Androsta-5 , 16-dien-3/3-ol

(24) in 475 cc of toluene and 75 cc of cyclohexanone was distilled (ca. 50 cc of distillate was collected) to eliminate moisture, 5 g of Al (0Pr<i> )3 in 50 cc of toluene was added and the solution was refluxed for 1 hour. Water then was added, volatile components were removed by steam distillation and the residue was extracted with chloroform. Evaporation of the dried extract, followed by crystallization of the residue from chloroform-hexane, yielded 7.53 g of Androsta-4, 16-dien-3-one (25). Another 0.97 g (total, 8.5 g, 86%) was obtained by chromatography of the mother liquor on neutral alumina.

EXAMPLE 8 Synthesis of Estra-1, 3 , 5 (10) , 16-tetraen-3-ol (28) The following method of synthesis is depicted in FIG. 6:

Estrone p-Toluenesulfonylhydrazone (27) Estrone (26) (270 g, 1.00 mole) and p-toluenesulfonylhydrazide (232.8 g, 1.25 mole) in dry methanol (2.5 liters) were heated under reflux for 20 hours. The mixture was transferred to a conical flask and allowed to cool. The crystalline product was filtered off under suction and washed with methanol (300 ml) . Further crops of product were obtained by sequentially evaporating the filtrate to 2000 ml, 800 ml and 400 ml, and allowing to crystallize each time. Total yield was 433.5 g (99%) .

1,3, 5 (10) ,16-Estratetraen 3-ol (28):

Estrone p-toluenesulfonylhydrazone (27) (219.0 g, 500 m mole) in dry tetrahydrofuran (8.0 liters) was cooled in a sodium chloride/ice bath. The mixture was mechanically stirred while n-butyl lithium (800 ml of a 2.5M solution in hexane, 2.00 mole) was added via double-ended needle. The mixture was stirred at room temperature for 3 days. Ice (250 g) was added, followed by saturated ammonium chloride solution (500 ml) . The phases were mixed by stirring and then allowed to settle. The aqueous phase was removed via aspiration with teflon tube and extracted with ether (500 ml) . The two organic phases were sequentially washed with the same batch of saturated sodium bicarbonate solution (500 ml) followed by saturated sodium chloride solution (500 ml) . The organic layers were dried (MgS04) and evaporated in vacuo to give crude product. This was subjected to flash filtration on 500 g silica gel 60, 230-400 mesh, eluting with ethyl acetate/hexane (25:75, 2.5 liters). The filtrate was evaporated in vacuo to give crystalline material. The product was recrystallized from methanol (300 ml) /water (75 ml) washing with methanol (80 ml)/ water (20 ml) . Further recrystallization from ethyl acetate/hexane (12.5:87.5) gave pure product (88.9 g, 70%) .

EXAMPLE 9

FIXATIVE EFFECT OF ESTRATETRAENOL Three 10% solutions of cineole in ethanol were prepared, containing, respectively, 0.1%, 1.0% and 10.0% by weight of estra-1 3 , 5 (10) , 16-tetraen-3-ol . Each sample was placed on a perfurmer's smelling strip and allowed to evaporate. The time required to fall below olfactory threshold was measured;

CONC. OF ESTR. TIME TO NON-DETECTION 0% (control) 5 minutes 0.1% 10 1.0% 25 10.0% 360

EXAMPLE 10 FIXATIVE EFFECT OF ANDROSTADIENONE Four solutions were prepared containing the same level of fragrances as the product Realm™. The samples contained respectively, 8 μg/ml, 12 μg/ml, 16 μg/ml and 0 (control) of androsta-4 , 16-dien-3-one . In a double blind study, the samples were placed on human skin and testers were asked to estimate the length of time the fragrances could be detected by smell. The sample containing 16 μg/ml of the androstadienone lasted the longest according to all testers.

It will be apparent to those skilled in the art that the objects of this invention have been achieved by providing the compositions described herein. Various changes may be made in the structure of the pheromones and in the compositions containing pheromones without departing from the concept of the invention. Further, features of some compositions disclosed in this application may be employed with features of other compositions. Therefore, the scope of the invention is to be determined by the terminology of the following claims and the legal equivalents thereof.

What is claimed is:

Claims

1. A method of fixing fragrances in a non-therapeutic fragrance composition comprising an odorant, comprising the step of adding to said composition an effective amount of a human pheromone sufficient to prolong the presence of the fragrance on the skin, said pheromone selected from the group consisting of 16 -Androstene steroids having the formula:

where Rl is selected from the group consisting of oxo, alpha -( beta -) hydroxy, alpha -( beta -) acetoxy, alpha -( beta -) propionoxy, alpha -( beta -) methoxy, alpha -( beta -) lower acyloxy, alpha -( beta -) lower alkyloxy, and alpha -( beta -) benzoyloxy; R2 is selected from the group consisting of hydrogen, hydroxy, acyl, acyloxy, alkoxy, methyl, hydroxymethyl, acylmethyl, acyloxymethyl, alkoxymethyl , lower alkyl, hydroxyalkyl, acylalkyl, acyloxyalkyl , and alkoxylalkyl ; and "a" and "b" are alternative sites for an optional double bond; and an Estrene steroid which has the formula:

wherein R4 is selected from the group consisting of hydrogen and alkyl, R6 is selected from the group consisting of hydrogen, lower alkyl, benzoyl, cypionyl, acetyl, glucuronide, lower acyl and sulfate; and "c" is an optional double bond.

2. The method of claim 1 wherein said pheromone is selected from the group consisting of 4, 16-Androstadien-3-one, 1, 3 , 5 (10) , 16-Estratetraen-3-ol and mixtures thereof .

3. The method of claim 1 wherein the concentration of said pheromone in the fragrance composition is at least about 50 ng/ml .

4. A method according to claim 1 wherein said fragrance composition comprises at least one human pheromone selected from the group consisting of 16 -Androstene steroids having the formula: I

where Rl is selected from the group consisting of oxo, alpha -( beta -)hydroxy, alpha -( beta -) acetoxy, alpha -( beta -) propionoxy, alpha -( beta -)methoxy, alpha -( beta -) lower acyloxy, alpha -( beta -) lower alkyloxy, and alpha -( beta -) benzoyloxy; R2 is selected from the group consisting of hydrogen, acylmethyl, acyloxymethyl, alkoxymethyl, lower alkyl, hydroxyalkyl , acylalkyl, acyloxyalkyl, and alkoxylalkyl; and "a" and "b" are alternative sites for a double bond.

5. A method according to claim 1 wherein said fragrance composition comprises at least one human pheromone selected from the group of Estrene steroids having the formula: II

wherein R4 is selected from the group consisting of hydrogen, and alkyl; R6 is selected from the group consisting of hydrogen, lower alkyl, benzoyl, cypionyl, acetyl, glucuronide, lower acyl and sulfate; and "c" is an optional double bond.