ESTRENE STEROIDS AS NEUROCHEMICAL INITIATORS OF CHANGE IN HUMAN HYPOTHALAMIC FUNCTION AND RELATED PHARMACEUTICAL COMPOSITIONS AND METHODS

Abstract: The invention relates to a method of altering hypothalamic function in an individual. The method comprises nasally administering a human semiochemical, e.g. an Estrene steroid, or a pharmaceutical composition containing an Estrene steroid, such that the ligand semiochemical binds to a specific neuroepithelial receptor. The steroid is preferably administered in the form of a pharmaceutical composition containing one or more pharmaceutically acceptable carriers. Other embodiments of the invention include pharmaceutical compositions containing the steroids.

Description: ESTRENE STEROIDS AS NEUROCHEMICAL INITIATORS OF CHANGE IN HUMAN HYPOTHALAMIC FUNCTION AND RELATED PHARMACEUTICAL COMPOSITIONS AND METHODS BACKGROUND Technical Field This invention relates generally to pharmaceutical compositions and methods for effectuating change in human hypothalamic function, thereby altering certain behavior and physiology mediated by the hypothalamus of individuals. More particularly, the invention relates to the use of certain Estrene steroids as neurochemical effectuators of physiology and behavior. Description of the Related Art The present invention relates to certain compounds, namely Estrene steroids and related compounds as will be described herein, and methods of using these compounds as human semiochemicals in order to alter hypothalamic function, thereby affecting certain consequent behavior and physiology, e.g. the reduction of anxiety. Estrene steroids are typified by 17β-Estradiol (1,3 ,5(10) -Estratriene- 3,17β-diol) , and are characterized by 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 pheromone properties of some Estrene steroids for some mammalian species has been described. Michael, R.P. et al.. Nature (1968) 218:746 refers to Estrogens (particularly Estradiol) as a pheromonal attractant of male rhesus monkeys. Parrot, R.F., Hormones and Behavior (1976) 2:207-215, reports Estradiol benzoate injection induces mating behavior in ovariectomized rats; and the role of the blood level of Estradiol in make sexual response (Phoenix, C.H., Phvsiol. and Behavior (1976) .16:305-310) and female sexual response (Phoenix, CH. , Hormones and Behavior (1977) 8.:356-362) in Rhesus monkeys has been described. On the other hand, there is little agreement in the literature as to whether or not pheromones as such play any role in the reproductive behavior and interpersonal communication of mammals (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). An embodiment of the subject invention concerns the non-systemic, nasal administration of certain Estrene steroids to affect a specific behavioral or physiological response in human subjects, e.g. a reduction of negative affect, mood, and character traits. In particular, nasal administration provides for contacting neuroreceptors of a heretofore poorly understood neuroendocrine structure, commonly known as the vomeronasal organ ("VNO"; also known as "Jacobson's organ"), with one or more steroid(s) or with compositions containing the steroid(s) . This organ is accessed through the nostrils of most higher animals - from snakes to humans, and has been associated, inter alia, with pheromone reception in certain species (see generally Muller-Schwarze & Silverstein, Chemical Signals. Plenum Press, New York (1980)). The axons of the neuroepithelia of the vomeronasal organ, located supra palatial, form the vomeronasal nerve and have direct synaptic connection to the accessory olfactory bulb and indirect input from there to the cortico- medial amygdaloid forebrain and hypothalamic nuclei of the brain. The distal axons of the terminal is nerve neurons may also serve as neurochemical receptors in the VNO. Stensaas, L.J., et al.. J. Steroid Biochem. and Molec. Biol. (1991) 39:553. This nerve has direct synaptic connection with the hypothalamus. Johnson, A. et al. (J. Otolarvngology (1985) 14.:71-79) report evidence for the presence of the vomeronasal organ in most adult humans, but conclude that the organ is probably non-functional. Contravening results which suggest that the VNO is a functional chemosensory receptor are reported by Stensaas, L. , et al.. supra; and by Moran, D.T., et al. , Garcia-Velasco, J. and M. Mondragon; Monti- Bloch, L. and B. Grosser - all in J. Steroid Biochem. and Molec. Biol. (1991) 39. It is apparent that it would be desirable to identify and synthesize human semiochemicals and pheromones and to develop pharmaceutical compositions and methods of use to influence hypothalamic function. This invention relates to the unexpected discovery that, when nasally administered to human subjects, certain neurochemical ligands, particularly certain Estrene steroids and related compounds, or pharmaceutical compositions containing certain Estrenes or related compounds, specifically bind to chemoreceptors of nasal neuroepithelial cells and this binding generates a series of neurolophysiological responses resulting in an alteration of hypothalamic function of an individual. When properly administered, the effect of certain of these compounds on the hypothalamus affects the function of the autono ic nervous system and a variety of behavioral or physiological phenomena which include, but are not limited to the following: anxiety, premenstrual stress, fear, aggression, hunger, blood pressure, and other behavioral and physiological functions normally regulated by the hypothalamus. Otto Appenzeller. The Autonomic Nervous System. An introduction of basic and clinical concepts (1990); Korner, P.I. Central nervous control of autonomic cardiovascular function, and Levy, N.M. and Martin, P.J. Neural control of the heart, both in Handbook of Physiology; Section 2: Cardiovascular System - the heart, Vol I, Washington DC, 1979, American Physiological Society; Fishman, A.P., et al. editors, Handbook of Physiology. Section 3: Respiratory System. Vol. II. Control of breathing. Bethesda MD. 1986. American Physiological Society. In some instances a single Estrene steroid, or related compound, is administered, in some instances combinations of Estrene steroids and/or related compounds are administered and in some instances one or more Estrene steroids are co- administered along with one or more Androstane steroids or a related compound. SUMMARY OF THE INVENTION Accordingly, it is an object of this invention to provide pharmaceutical compositions which contain human semiochemicals or pheromones and are suitable for nasal administration in an individual. It is also an object of this invention to provide methods of using these compositions to alter hypothalamic function of an individual. It is a further object of this invention to provide methods of using these compositions to affect physiological and behavioral functions of individuals which are normally regulated by the hypothalamus. Finally, it is an object of this invention to provide methods of altering hypothalamic function which have the following advantages: 1) administration directly to the chemoreceptors in the nasal passage and the vomeronasal organ, without pills or needles - i.e., non-invasively; 2) a mode of drug action through the nervous system and not through the circulatory system - thus brain function can be affected without consideration of the blood- brain barrier; 3) a direct means of affecting the hypothalamus - there is only one synaptic junction between pheromone receptors and the hypothalamus; and, 4) providing a highly specific drug effect, thereby greatly reducing the potential for undesirable side-effects - this because sensory nerves are addressed to a specific location in the brain. Additional objects, advantages and novel features of the invention will be set forth in part in the description which follows, and in part will become apparent to those skilled in the art upon examination of the following, or may be learned by practice of the invention. Objects of this invention are achieved by providing a pharmaceutical composition suitable for nasal administration in an individual. The composition contains a pharmaceutically acceptable carrier and an Estrene steroid with the formula: wherein R, is selected from the group consisting essentially of one or two hydrogen atoms, methyl, methylene, and one or two halo atoms; R 2 is absent or is selected from the group consisting essentially of hydrogen and methyl; R 3 is selected from the group consisting essentially of oxo, hydroxy, lower alkoxy, lower acyloxy, benzoyl, cypionyl, glucuronide and sulfonyl; R 4 is selected from the group consisting essentially of hydrogen, hydroxy, lower alkoxy, lower acyloxy and halo; R 5 is absent or is selected from the group consisting essentially of hydrogen, hydroxy, lower alkoxy and lower acyloxy; R^ is a hydrogen or a halo; and "a" represents optional aromatic unsaturation of ring A of said steroid, or "b", "c", and "d" are each optional double bonds; and "e", "f" , "g", "h", "i" and "j" are each optional double bonds. In this embodiment, the steroid is preferrably administered in the form of a pharmaceutical composition containing one or more pharmaceutically acceptable carriers. A preferred class of compounds are those in which "a" is present and "g", "h" or "i" are optional double bonds. The class wherein "h" and "i" are both double bonds is also preferred. Another preferred class contains "b", "c" or "j" as a double bond. Yet another class contains "c" and "d" as double bonds. Still another class contains R 2 as methyl and (1) "e" as a double bond, (2) R, is methylene or a single hydrogen, or (3) "f" is a double bond. The term lower alkyl, lower alkoxy, etc., encompasses carbon chains of l to 6 carbon atoms, preferrably 1 to 4 carbon atoms. Halo includes I, Br, F and Cl. Other objects of this invention are achieved by providing a method of altering hypothalamic function and autonomic function in an individual. A ligand for a chemoreceptor displayed on the surface of a nasal neuroepithelial cell is provided wherein the cell is a part of tissue other than olfactory epithelia; and, the ligand is administered within a nasal passage of the individual such that the ligand binds specifically to the chemoreceptor, resulting in an alteration of hypothalamic function of the individual. All embodiments of this application relate to and include the functional equivalents of the steroid structures disclosed in these embodiments and to those modified steroids which demonstrate said functional equivalence, whether or not the modified steroids are explicitly disclosed. Brief Description of the Drawings Figure 1 illustrates the synthesis of 1, 3 ,5 (10) , 16-Estratetraen-3-ol. Figures 2A, 2B, and 2C are graphic representations of the electrophysiological effect on receptor potential of the localized administration of particular steroids to the vomeronasal organ of male subjects (Fig. 2A) and to the olfactory epithelium (Fig. 2C) . Figure 2B is a graphic comparison of the effect of an Estrene on the VNO receptor potential of male and female subjects. Figure 3 is a graphic representation of the electrophysiological effect of the localized administration of particular steroids to the vomeronasal organ of male (3A) and female (3B) subjects. Figure 4 depicts various autonomic responses of male subjects to 1,3,5(10), 16-Estratetraen-3-yl acetate. A = receptor potential of the vomeronasal neuroepithelium; B = change in galvanic skin response (K-ohms) ; C = change in skin temperature (degrees C.) . Figure 5 depicts comparative changes in potential of the VNO after exposure to the methyl ether and the acetate of 1,3,5(10) ,16-Estratetraen-3- ol. Figure 6 depicts sexual dimorphism in local and autonomic responses to the stimulation of the VNO with vomeropherins. Various vomeropherins (200 fmoles) and the diluent control were administered to 30 male and 30 female subjects (ages 20 to 45 ) as described. Bars indicate the mean response of the population. Figs. 6A & B: EVG responses were measured as described in male (A) and female (B) subjects. Figs. 6C & D: Electrodermal activity was measured as described. Changes (measured in κΩ) in response due to delivery of vomeropherins to the VNO of each subject are shown in male (C) and female (D) subjects. Figs. 6E & F: Alpha-cortical activity was measured as described. Changes in response due to delivery of vomeropherins to the VNO of male (E) and female (F) subjects. Figs. 6G & H: Skin temperature (ST) wad measured as described. Changes in response due to delivery of vomeropherins to the VNO of each subject are shown in male (G) and female (H) subjects. A = 1, 3, 5(10) ,16-Estratetraen-3-yl acetate B = Androsta-4,16-dien-3-one C = 1, 3 , 5 (10) , 16-Estratetraen-3-ol D = 3-Methoxy-Estra-l, 3 , 5(10) , 16-tetraene E = Androsta-4 , 16-dien-3α-ol F = Androsta- 4, 16-dien-3B-ol Figure 7 depicts electro-olfactgrams of male and female subjects induced by stimulation of the OE with olfactants and vomeropherins A: 400 fmoles of the olfactants 1-carvone and cineole as well as 200 fmoles of the vomeropherins A, B, C, D and F; and the stereoisomer E were applied separately as one second pulses to the OE of 20 subjects (both male and female) and each EOG response was recorded as described. The olfactants as well as E and B produced significant (p<0.01) local response. B: 400 fmoles of the olfactants 1-carvone and cineole do not induce a significant EVG response when delivered to the VNO of male and female subjects. Figure 8 depicts the electrophysiological effect of the following vomeropherins on the vomeronasal organ of 20 female subjects: G = Androst-4-en-3-one H = Androsta-4 , 16-diene-3 , 6-dione J = 10, 17-Dimethylgona-4 , 13 (17) -dien-3-one K = 1,3,5 (10) , 16-Estratetraen-3-ol-methyl ether L = 1,3,5(10) ,16-Estratetraen-3-yl-propionate EVG = Electro-vomeronasogram GSR = Galvanic Skin Response = Electrodermal Activity (EDA) ST = Skin Temperature Figure 9 depicts the electrophysiological effect of vomeropherins on the vomeronasal organ of 20 male subjects. M = 1,3,5 (10) -Estratrien-3-ol Figure 10 depicts the synthesis of Estra- 1,3,5(10) ,6-tetraen-3-ol and Estra-4,16-dien-3-ol. Figure 11 depicts the synthesis of compounds described in Examples 16 through 19. Detailed Description of the Invention I. Definitions An "affect" is a transient feeling state. Typical negative affects are feelings of nervousness, tenseness, shame, anxiousness, irritability, anger, rage, and the like. "Moods" are longer lasting feeling states such as guilt, sadness, hopelessness, worthlessness, remorsefulness, misery, unhappiness and the like. "Character" traits are more permanent aspects of an individual's personality. Typical negative character traits are sensitivity, regretfulness, blameworthiness, stubbornness, resentfulness, bitterness, timidness, laziness and the like. "Androstane steroids" are aliphatic polycyclic hydrocarbons characterized by a four-ring steroidal structure, with a methylation at the 10- and 13- positions. An Androstene steroid is a subset of Androstanes, commonly understood to mean that the compound has at least one double bond. Commonly, unless a compound is described as a gonane it is understood that the compound has an 18- carbon group. However, it is intended that 18-Nor-Androstanes are herein regarded as Androstane steroids. Furthermore, all derivatives which have the structural characteristics described above are also referred to generically herein as Androstane steroids. "Estrene steroids", as the term is used herein, are aliphatic polycyclic hydrocarbons with a four-ring steroidal structure, at least one double bond in the A-ring, no methylation at the 10-position and an oxo, hydroxyl or hydroxyl derivative such as an alkoxy, ester, benzoate, cypionate, sulfate or glucuronide, at the 3-position. Derivatives which contain these structural characteristics are also referred to generically as Estrene steroids. Estrene steroids are also known as Estrogenic steroids in the parent application, U.S.S.N. 07/638,185, and these two terms are intended to be equivalent. The following structure shows the four-ring steroidal structure common to 16-Androstene and Estrene steroids. In describing the location of groups and substituents, the following numbering system will be employed: "Sexually dimorphic" refers to a difference in the effect of, or response to, a pharmaceutical agent between males and females of the same species. An "effective amount" of a drug is a range of quantity and/or concentration which brings about a desired physiological and/or psychological effect when administered to a subject in need of the drug. In the present case, a needy subject is one in need of hypothalamic modulation or regulation, or a subject in need of alteration of a physiological or behavioral characteristic normally affected by the hypothalamus. The effective amount of a given drug may vary depending upon the route of administration. For example, when the steroid is administered as a solution applied to the facial skin of a subject an effective concentration is from about 1 to about 100 μg/ml, preferably about 10 to about 50 μg/ml and most preferably about 20 to about 30 μg/ml. When the steroid is introduced directly into the VNO an effective amount is about 1 pg to about 1 ng, more preferably about 10 pg to about 50 pg. When the steroid is administered to the nasal passage by ointment, cream, aerosol, or the like, an effective amount is about 100 pg to about 100 micrograms, preferably about 1 ng to about 10 micrograms. It follows that some drugs may be effective when administered by some routes, but not effective when administered by other routes. The "hypothalamus" is the portion of the diencephalon comprising the ventral wall of the third ventricle below the hypothalamic sulcus and including structures forming the ventricle floor, including the optic chiasma, tuber cinereum, infundibulum, and mammillary bodies. The hypothalamus regulates the autonomic nervous system and controls several physiological and behavioral functions such as the so-called fight and flight responses, sexual motivation, water balance, sugar and fat metabolism, hunger, regulation of body temperature, endocrine secretions, and others. The hypothalamus is also the source of vasopressin which regulates blood pressure, and oxytocin which induces parturition and milk release. All hypothalamic functions are potentially modulatable by the semiochemical therapy described herein. A "ligand", as used herein, is a molecule which acts as a chemical signal by specifically binding to a receptor molecule displayed on the surface of a receptor cell, thereby initiating a signal transduction across the cell surface. Binding of ligands to chemosensory receptors can be measured. Chemosensory tissue, such as vomeronasal neuroepithelium or olfactory neuroepithelium, contains a multiplicity of neuroreceptors cells, each displaying at least one cell surface receptor. Many of the receptor molecules have identical ligand specificity. Therefore, when the tissue is exposed to a ligand for which it has specificity (for example a exposure of the VNO to a semiochemical) a summated change in cell surface receptor potential can be measured. As used herein, "lower alkyl" means a branched or unbranched saturated hydrocarbon chain of 1 to 4 carbons, such as, for example, methyl, ethyl. n-propyl, i-butyl and the like. "Alkoxy" as used herein is used in its conventional sense to mean the group -OR where in R is an alkyl as defined herein. A "pheromone" is a substance that provides 5 chemical means of communication between members of the same species through secretion and nasus reception. In mammals pheromones are usually detected by receptors in the vomeronasal organ of the nose. Commonly, pheromones effect development, reproduction 0 and related behaviors. A "semiochemical" is a more general term which includes pheromones and describes a substance from any source which functions as a chemosensory messenger, binds to a specific neuroepithelial receptor, and induces a physiological 5 or behavioral effect. A "vomeropherin" is a semiochemical whose physiologic effect is mediated through the vomeronasal organ. A picogram (pg) is equal to .001 nanograms (ng) . A ng is equal to .001 microgram (μg) . A μg is 0 equal to .001 mg. II.. Modes for Carrying Out the Invention A. Estrenes Useful in the Invention The invention is directed in part to certain Estrene steroids which are structurally related to Estradiol (also referred to as 1,3,5(10)-Etratriene-' 3,176-diol). Steroids within the group are characterized by a aromatic 1,3,5(10) A-ring and a hydroxyl or hydroxyl derivative at the 3-position. Estrenes especially suitable for use in the present invention include those where, independently, R j = oxo, 6- hydroxy, hydrogen; R 2 = α- hydroxy, 6- hydroxy, hydrogen; R 3 = hydrogen, lower alkyl, acetyl, propionyl; and "a" is or is not a double bond (as depicted in formula I) . Preferred estrenes include 1,3,5(10) - Estratriene-3,17β-diol; 1,3,5(10)-Estratriene- 3,16α,176-triol; 1,3,5(10)-Estratriene-3-ol-17-one; 1,3,5(10) ,16- Estratetraen-3-ol; 1,3,5(10), 16- Estratetraen-3-ol methyl ether; and 1,3,5(10) ,16- Estratetraen-3-yl-propionate; 1,3,5(10) ,16- Estratetraen-3-yl acetate. Most of these steroids and their glucuronide, sulfate, cypionate, and benzoate derivatives, are compounds known in the art and are commercially available, e.g., from Sigma Chemical Co., Aldrich Chemical Co. , etc. Alkoxy derivatives and their synthesis are also known in the art and taught in U.S. Patent No. 2,984,677, herein incorporated by reference. 1,3,5(10) ,16-Estratetraen-3-ol is available from Research Plus, Inc. and from Steraloids, Inc. Preparation of the acetate and propionate derivatives of this compound are described herein. Chart 1 includes estrenes to which the invention is directed, but do not limit its scope. The synthesis diagrams that follow depict intermediate and substructure syntheses for the preparation of these estrenes: CHART I: ESTRANES SUBSTRUCTURE SYNTHESES Referring to the preceding table, the following are exemplary syntheses for intermediates in a given row (El through E12) or column (Nl through N4) . Type E El : H .do H (Methyl ether of E2) .d (E1o) E2 : Commercially available substructure , for example , ESTRONE . E3 : H (E1) (E3) James R. Bull and Jan Floor, J. Chem. Soc. Perkin I, 1977 (7), 724. E4 Commerically available substructure, for example, 6-DEHYDROESTRONE. E5 : .CO 1. H * OO 2. PyHBr 3 V. I. Mel'nikova and K. K. Pivnitskii, Zhurnal Organickeskoi Khisnii, 1974, Vol. 10, No. 5, pp. 1014-1019). E6: (Acetate of E2) (Acetate of E6) Hidetoshi Takagi, Ken-ichi Komatsu, and Itsuo Yoshisawa, Steroids, 1991, Vol. 56, p. 173. E7: Michel Mauney and Jean Rigaudy, Bull. Soo. Chien, 1976, No. 11-12, 2021. E8: (Methyl ether of E2) K. J. San, R. H. Blank, R. H. Evans, Jr., L. I. Feldman, and C. E. Holmbund, J. Org. Chem., 1964, 29, 2351. E9: Commercially available substructure, as in EQUILIN. E10: Commercially available substructure, as in EQUILENIN. Ell: (E8) (E11) A. N. Cherkasov, A. M. Ponomarev, and K. K. Pivnitskii, Zhurnal Organiskeskoi Khimii, 1971, Vol. 7, No. 5, pp. 940-947. E12: OH (Methyl ether of EG) (Methyl ether of El 2) Hidetoshi Takagi, Ken-ichi Komatsu, and Itsuo Yoshisawa, Steroids, 1991, Vol. 56, p. 173. Type N Nl : n-BuLi (N1) N2 : 1. Robert H. Shapiro and Carl Djerassi, J. Am. Chem. Soc, 1964, 86., 2825. 2. Pilar Lupόn, Frances C. Canals, Arsenio Iglesias, Joan C. Ferrer, Albert Palomar, and Juan-Julio Bonet, J. Org. Chem. 1988, 53_, 2193-2198. N3: 1. Gunther Drefahl, Kurt Ponold and Hans Schick, Berichte, 1965, £8 / 604. 2. Richard H. Peters, David F. Crows, Mitchell A. Avery, Wesley K. M. Chong, and Masako Tanabe, J. Med. Chem., 1989, 3_2, 1642. N4; COLLIDINE (N4) 1. Franz Sonheimer, 0. Moncera, M. Viquiza & G. Rosenkranz (1955) J. Am. Chem. Soc. 77:4145. 2. William F. Johns, J. Org. Chem., 1961, 2ϋ, 4583. Methylestrenes Harold J. Nicholas, J. Org. Chem., 1958, 22, 1747, Richard H. Peters, David F. Crows, Mitchell A. Avery, Wesley K. M. Chong, and Masako Tanabe, J. Med. Chem., 1989, 3_2, 1642. (RACEMIC) M. B. Green and F. J. Zeelen, Tetrahedron Letters, 1982, Vol. 23, No. 35, pp. 3611-3614. Synthesisable compounds therefore include these, together with those derived from them; i.e., 17-Methyl-Nl, 173-Methyl-N2, or 14α-Methyl-N4, incombination with El, E2, E3, E5, E6, E7, E8, Ell or E12. Haloestrenes George A. Boswell in patent CA. 70:58140g, following. G. Michael Blackburn, Brian F. Taylor, and Andrew F. Worrall, Journal of Labelled Compounds and Radiopharmaceuticals, 1986, Vol. XXIII, No..2, p. 197. Synthesisable compounds therefore include these, together with those derived from them; i.e., 17-Fluoro-Nl in combination with El, E2, E3, E5, E6, E7, Ell or E12. In addition, 17-Iodo-Nl in combination with E2, E6 or E12. B. Androstenes Useful in the Invention The invention is additionally directed to compositions and methods involving the combination of the aforementioned Estrene steroids with certain Androstane steroids, preferably Androstane steroids with the formula: wherein P is selected from the group consisting of oxo, -(β-) hydroxy, α-(β-) acetoxy, α-(β-) propionoxy, -(β-) methoxy, α-(β-) lower acyloxy, α-(β-) lower alkyloxy, and α-(β-) benzoyloxy; P 2 is selected from the group consisting of methyl, hydroxymethyl, acyloxymethyl, alkoxymethyl, lower alkyl, hydroxyalkyl, acyloxyalkyl, and alkoxylalkyl; P 3 is absent or is selected from the group consisting of methyl, hydroxymethyl, acyloxymethyl, alkoxymethyl, lower alkyl, hydroxyalkyl, acyloxyalkyl, and alkoxylalkyl; P is selected from the group consisting of hydrogen, oxo, halo, hydroxy, alkoxy, and acyloxy; P 5 represents one or 2 substituents, wherein P 5 comprises one or two hydrogen atoms, methyl, methylene, or one or two halo atoms; P 6 is hydrogen or halo; and "a HIb-,n", and are alternative sites for optional double bonds. One class of preferred steroids has "b" as a double bond, particularly wherein "c" or "d" is also a double bond. Another preferred class has "a" and 11 c" as double bonds. Yet another preferred class contains P 3 as a methyl group, "h" as an optional double bond, and P 5 as methylene or one or two hydrogen atoms. A class of steroids wherein "a" or "b" is a double bond is also preferred. Preferred steroids include Androsta-4, 16- dien-3-one (P j = oxo, a = double bond, P 2 = methyl, commercially available from Steraloids, Inc.), Androsta-4, 16-dien-3α-ol (P]= α-OH, a = double bond, P 2 = methyl), and Androsta-4, 16-dien-36-ol (P ! = 6- OH, a = double bond, P 2 = methyl) , syntheses of which are described in the commonly owned, co-pending application, entitled "Androstane Steroids as Neurochemical Initiators of Change in Human Hypothalamic Function and Related Pharmaceutical Compositions and Methods", U.S.S.N. presently unassigned (a continuation-in-part of U.S. Patent Application Serial No. 07/903,604, filed 24 June 1992) , and incorporated by reference herein. C. Synthetic Methods 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. Alkoxy derivatives of Estrenes are prepared from their corresponding hydroxy steroids by reaction with an alkylating agent such as trimethyloxoniu 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. Alternatively, a base such as K 2 C0 3 may be used in solvents such as ethanol or acetone. Purification of the products is accomplished by means of chromatography and/or crystallization, as known to those skilled in the art. D. Pharmaceutical Compositions and Methods of Use An embodiment of the subject invention is a method of altering the hypothalamic function of an individual. Another embodiment is altering an autonomic function of an individual. These autonomic functions include, but are not limited to, heart rate, respiratory rate, brain wave patterns (percentage alpha cortical activity), bodytemperature. Other embodiments include, but are not limited to, methods of diminishing negative affect, negative mood or negative character traits of an individual. Another embodiment is a method of treating female premenstrual stress. All of these embodiments are accomplished by means of the non- systemic, nasal administration of certain Estrene steroids, combinations of Estrene steroids and combinations of one or more Estrene steroids and one or more Androstane steroids. This particular mode of administration is distinguished from alternative modes, such as ingestion or injection, in several important ways, these by virtue of the direct contact with the VNO provided by the nasal administration of the steroid ligand. In the methods of this invention, the appropriate ligand is administered directly to the chemoreceptors in the nasal passage and the vomeronasal organ, without pills or needles - i.e., noninvasively. Drug action is mediated through binding of the ligands, described herein, to specific receptors displayed by neuroepithelial cells in the nose, preferably in the VNO. This mode of drug action is through the nervous system and not through the circulatory system - thus brain function can be affected without consideration of the blood-brain barrier. These methods of treatment provide a direct means of affecting the hypothalamus through the nervous system because there is only one synaptic junction between pheromone receptors and the hypothalamus. Because sensory nerves are addressed to a specific location in the brain, this method has a highly specific drug effect> thereby greatly reducing the potential of undesirable side-effects. VNO contact is important because the VNO is associated with chemoreceptive/pheromonal function. The VNO consists of a pair of blind tubular diverticula which are found at the inferior margin of the nasal septum. The VNO contains neuro-epithelia, the axons of which have direct synapses to the amygdala and from there, to the hypothalamus. The existence of the VNO has been well documented in most terrestrial vertebrates including the human, fetus; however, in adult humans it is generally thought to be rudimentary (See Johnson, et al. , supra) . The active compounds described herein, or their sulfated, cypionated, benzoated, propionated, halogenated or glucuronated derivatives, may be administered directly, but are preferably administered as compositions. They are prepared in a liquid dosage form such as, for example, liquids, suspensions or the like, preferably in unit dosage forms suitable for single administration of precise dosages. Liquid dosages may be administered as nose drops or as an aerosol. Alternatively, the active compound can be prepared as a cre e or an ointment composition and applied topically within the nasal cavity. As another alternative, delivery may occur by controlled release of these agents by encapsulation either in bulk or at a microscopic level using synthetic polymers, such as silicone, and natural polymers such as gelatin and cellulose. The release rate can be controlled by proper choice of the polymeric system used to control the diffusion rate (Langer, R.S. and Peppas, N.A. , Biomaterials 2,201, 1981). Natural polymers, such as gelatin and cellulose slowly dissolve in a matter of minutes to hours while silicone remains intact for a period of months. The compositions will include a conventional pharmaceutical carrier or excipient, one or more of the active Estrene compound(s) of Formula I, and the composition may or may not additionally include one or more Androstane steroids. In addition, the compositions may include other medicinal agents, pharmaceutical agents, carriers, adjuvants, etc. 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 5α- Androst-16-en-3α-ol and 5α-Androst-16-en-3-one, 4,16- Androstadien-3-one, 5α-Androstadien-3β-ol, and perhaps 5α-Androstadien-3α-ol, are naturally occurring in humans and may be present on the skin. It is estimated that the naturally occurring maximum concentration of a 16- Androstene steroid on human skin is from 2 to 7 ng/cm 2 . During intimate contact it is estimated that a human would be exposed to no more than 700 ng of a naturally occurring steroid. Since these compounds are relatively non-volatile, it is estimated that, even during intimate contact, a human subject would inhale no more than 0.7 pg of a naturally occurring steroid from the skin of another. From the amount inhaled only about 1% would reach the receptors of the vomeronasal organ. Thus the estimated maximum natural exposure to naturallyproduced pheromones would be 0.007 pg. The amount of active compound administered will of course, be dependent on the subject being treated, the severity of-the affliction, the manner of administration, the frequency of administration, andthe judgment of the prescribing physician. However, a single dosage of at least about 10 picograms, delivered directly into the lumen of the vomeronasal organ, is effective in eliciting a transient autonomic response, when administered to the nasal cavity, the dosage is about 100 picograms to about 100 micrograms, preferably about 1 nanogram to about 10 micrograms, more preferably about 10 nanograms to about 1 microgram. The frequency of administration is desirably in the range of an hourly dose to a monthly dose, preferably from 8 times/day to once every other day, more preferably 1 to 3 times per day. Ointments containing one or more active compounds and optional pharmaceutical adjuvants in a carrier, such as, for example, water, saline, aqueous dextrose, glycerol, ethanol, and the like, can be prepared using a base such as, for example, petroleum jelly, lard, or lanolin. Liquified pharmaceutically administrable compositions can, for example, be prepared by dissolving, dispersing, etc. an active compound as defined above and optional pharmaceutical adjuvants in a carrier, such as, for example, water, saline, aqueous dextrose, glycerol, ethanol, and the like, to thereby form a solution or suspension. If desired, the pharmaceutical composition to be administered may also contain minor amounts of nontoxic auxiliary substances such as wetting or emulsifying agents, pH buffering agents and the like, for example, sodium acetate, sorbitan monolaurate, triethanolamine sodium acetate, triethanolamine oleate, etc. Actual methods of preparing such dosage forms are known, or will be apparent, to those skilled in this art; for example, see Remington's Pharmaceutical Sciences. Mack Publishing Co., Easton, PA, 15th Ed., 1975. The composition or formulation to be administered will, in any event, contain a quantity of one or more of the active compound(s) in an amount effective to alleviate the symptoms of the subject being treated. For aerosol administration, the active ingredient is preferably supplied in 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 monooleate) 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. Yet another means of administration is topical application of a volatile liquid composition to the skin, preferably facial skin, of an individual. The composition will usually contain an alcohol such as ethanol or isopropanol. A pleasant odorant may also be included in the composition. E. Measuring Affect. Mood and Character Traits. Feeling states associated with affects, moods and character traits are generally measured by use of a questionnaire. For example questionnaires comprising a number of adjectives which refer to feeling states may be administered to an individual. The individual evaluates his or her feeling state described by the adjective and rates the intensity of the feeling on a numerical scale. Clustering of related adjectives and statistical analysis of a subject's evaluation of each adjective provides a basis for the measurement of various feeling states. Alternatively, feeling states may be measured by autonomic changes, such as those used in polygraphic evaluations (galvanic skin response, pulse rate and the like) . Cabanac, M. Annual Review of Physiology (1975) 22:415; Hardy, J.D. , "Body Temperature Regulation", Chapter 59, pp. 1417. In: Medical Physiology. Vol. IIEd. : VB Mountcastle (1980) ; Wolfram Bouscein. Electrodermal Activity (Plenum Press 1992) . In addition, non-verbal cues such as facial expression and body posture may be evaluated. F. Uses in the Treatment of Certain Types of Psychiatric Disorders. Compositions suitable for topical and/or 10 intranasal administration, and which contain one or more of the steroidal substances described herein, are useful as therapeutics with efficacy in the treatment of certain types of psychiatric disorders, particularly certain types of neuroses. In a preferred embodiment, and unlike other drug formulations designed to invoke a systemic effect, the composition is formulated to minimize the likelihood of mucosal or transdermal absorption since the mode of action of these substances as therapeutics is by stimulation of neural receptors in the nose, more particularly in the VNO. Since the compositions of the instant invention are effective as the result of stimulation of chemosensory receptors in the VNO and not as a result of systemic circulation, absorption and the attendant incorporation of the active ingredient or ingredients into systemic circulation would not be efficacious, but rather might actually increase the likelihood of side effects. These compositions are useful when administered in a method such that one or more of the Estrenes or 16-Androstenes of the composition are provided to the VNO of the subject. Treatable neuroses may be acute and/or transient or they may be persistent or recurrent. They generally involve abnormal symptoms that may include mood changes (anxiety, panic, depression) or limited abnormalities of thought (obsessions, irrational fears) or of behavior (rituals or compulsions, pseudoneurological or hysterical conversion signs) . In such disorders, these compositions may have some beneficial effects for short periods, particularly by modifying associated anxiety or depression or the like. Other characterological disorders may also be treatable by the intranasal administration of the compositions of this invention. These conditions include characteristic personality styles, e.g., paranoid, withdrawn, psychopathic, hypochondrial and the like; or behavioral patterns, e.g., abuse of alcohol or other substances, socially deviant or perverse behavior and the like that may run counter to societal expectations. Certain physical or physiological conditions, both normal (such as menstruation) and disease or injury related (such as chronic illness) may have a psychological component that manifests as a psychiatric disorder, neurotic disorder, or as an anxious or depressed affect or mood. These conditions are also treatable by methods of administration of the compositions of the present invention such that one or more of the estrenes or androstenes of the composition are provided to the VNO of the subject. i. Anxiety Anxiety is a disagreeable emotional state characterized by feelings of uneasiness, impending danger, apprehension or tension. Compositions suitable for intranasal administration, and which contain one or more of the Estrene steroid substances described herein, are useful as therapeutics with efficacy in the reduction of anxiety. There are numerous manifestations of this disorder ranging from mild perturbations to syndromes which can be incapacitating. Anxiety is not only a cardinal symptom of may psychiatric disorders but also a component of many normal physiological and social conditions. In addition, symptoms of anxiety are commonly associated with depression and especially with dysthymic disorder (neurotic depression) and many personality disorders. Anxiety disorders can be separated and include panic disorders, phobias and generalized anxiety disorders, post-traumatic stress disorder and obsessive-compulsive disorder. ii. Drugs used in the treatment of anxiety Drug treatment is the major treatment method of anxiety, often used in conjunction with behavior therapy. The drugs used most frequently to treat anxiety are the benzodiazepines. Benzodiazepines have hypnotic, sedative, anxiolytic, anticonvulsant and muscle relaxant actions. Consequently, they are used for may indications and in many different conditions than anxiety disorders (e.g. , insomnia, alcohol withdrawal states, muscle spasms, epilepsy, anesthesia and sedation for endoscopic procedures) . These drugs are relatively safe, and have advantages in comparison with the previously used barbiturates. They have a rapid onset of action, and produces a sense of euphoria. However, despite their good safety record, benzodiazepines are associated with a number of properties that limit their use including sedation, ataxia, memory impairment, reduced motor coordination and dangerous addictive effects, especially when used with alcohol. Both physicians and patients are increasingly aware of this potential for addiction; and there is an evident desire, if not trend, to move away from their longter use. Recently, a new class of non-benzodiazepine anxiolytic agents, azaspirodecanediones, which may have more selective anxiolytic properties, has been approved. The first marketed member of this class is buspirone. This compound has been reported to be as effective as benzodiazepines in the treatment of Generalized Anxiety Disorder. Compared to benzodiazepines, buspirone is less sedating, potentiates the effect of ethanol to a lesser degree, and has a low addictive potential. However, in normal clinical use, it has been found to be difficult to substitute buspirone for benzodiazepines in patients already receiving benzodiazepines since buspirone does not create a feeling of euphoria, and patients do not feel as satisfied as when taking benzodiazepines. In addition, buspirone's slow onset of action (1-3 weeks) limits its effectiveness in treating acute anxiety. Systemically administered buspirone anxiolytic has been shown to have a high affinity for the CNS serotonin 5-HT receptor and this is thought to be the mode of anxiolytic action. Another class of new anxiolytic is the systemically administered benzofuran derivatives. These substances demonstrate a high affinity for the CNS dopamine D2 receptor. iii. Mood (Affective) disorders The compositions of the instant invention are also useful in the treatment of some of the symptoms of certain types of mood disorders. Mood disorders such as major depression and mania are characterized by changes in mood as the primary clinical manifestation. Either extreme of mood may be associated with psychosis, characterized by disordered delusional thinking and perceptions, often congruent with the predominant mood. Conversely, psychotic disorders may have associated or secondary changes in mood. Similarly, normal grief, sadness and disappointment and the dysphoria or demoralization often associated with medical illness or with normal cyclic dysfunctions such as premenstrual stress may also be mitigated by the intranasal application of the compositions of the instant invention. iv. The mode of action of the compositions of the instant invention The mode of action of the compositions of the instant invention is quite different from any modes of treatment that have been heretofore reported. The active steroid and steroid-like compounds described in the instant invention stimulate and/or bind to receptors in the VNO that are directly available to topical administration. Direct access to CNS receptors is not required. Stimulation of these receptors then produces a signal transmitted through a neural pathway and inducing a neuropsychic response, likely in the hypothalamic region of the subject's brain. This response manifests as discrete changes in a variety of autonomic functions, including but not limited to pulse rate, respiratory frequency, E.E.G. patterns, evoked potential, skin temperature, galvanic skin response and pupillary diameter. The response also manifests as a measurable reduction of negative affect and negative mood. While the relationship between the modification of hypothalamic function and the treatment of psychiatric disorders is not fully understood, it is clear that the compositions of this invention are effective in the reduction of self-evaluative negativity - negative mood characteristics as well as negative character. These changes are accompanied by autonomic changes, effected by the hypothalamus, that are consistent with an increase in parasympathetic tone (or alternatively a decrease in sympathetic tone) . Since the active compounds of this invention are pheromone-like in their specificity, the compounds exhibit a species specificity in their stimulation and activity add therefore demonstration of efficacy cannot be done in animal models. III. Examples The following examples are intended to illustrate but not to limit the invention. Abbreviations used in the examples are as follows: aq.=aqueous; RT.=room temperature; PE=petroleum ether (b.p. 50-70°); DMF=N, N-dimethylformamide; DMSO- dimethyl sulfoxide; THF=tetrahydrofuran. Example 1 Synthesis of Estra-1.3.5(10) .16-tetraen-3-τ ol (28). The following method of synthesis is depicted in Figure 1: 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 2.5 M 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 (MgS0 4 ) 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 2 - Synthesis of Acyl derivatives of 1.3.5(10) . 16-Estratetraen-3-ol. To 1,3,5(10) ,16-Estratetraen-3-ol (254 mg, 1.00 mMole) in ether (10 ml) is added acetic anhydride (0.25 ml) (or propionic anhydride for the propionate) followed by pyridine (0.25 ml) and the mixture is stirred at room temperature for 16 hours. The mixture is poured into ice/water and extracted with ether (2 X 20 ml) . The organic extracts are washedwith water, saturated copper sulfate solution, water, and saturated sodium chloride solution, dried (MgS0 4 ) and evaporated in vacuo to give the crude material. This is purified by flash chromatography on 17.5 g silica gel 60 (230-400 mesh) eluting with 10%-12% ethyl acetate/hexane to give the pure product (192 mg, 65%) . Exa ple 3 - Synthesis of Estra-4.16-dien-3-one (1) . To estra-1, 3 , 5 (10) , 16-tetraene-3-methyl ether (551.5 mg., 2.055 mmol) in 8.6 ml of anhydrous THF, approximately 30 ml of anhydrous ammonia, and 6.76 g of t-butyl alcohol was added lithium wire (0.24 g, 35 mg- atom) cut in small pieces. The reaction mixture was refluxed 4 1/2 h under argon, after which methanol (2.3 ml) was added and the ammonia was allowed to boil off overnight. The residue was dissolved in 25 mL of methanol and was acidified to approximately pH 1 with 5N HCI. After heating in an oil bath between 55 and 70°C for 15 min. the cooled hydrolysis mixture was partitioned between 25 ml of water and 50 ml of ethyl acetate and the aqueous phase was extracted with 25 ml of ethyl acetate. The combined organic extracts were washed with 25 ml of saturated sodium bicarbonate and 25 mL of brine, dried over magnesium sulfate, and filtered. Removal of solvent under reduced pressure yielded 0.57 g of oily residue which was purified by flash chromatography on silica gel (eluent: 15% ethyl acetate/hexane) followed by recrystallization from pentane to give crystals (206.1 mg, 39%) homogeneous to TLC, m.p. 67-71°C. Example 4 - Synthesis of Estra-2 ,5(10) , 16-triene-3-methyl ether ( 2 ) . To Estra-l, 3 , 5 (10) , 16-tetraene-3-methyl ether (1.22 g, 4.54 mole) in 19 ml of anhydrous THF, 14.99 g of t-butyl alcohol, and approximately 70 ml of anhydrous ammonia was added lithium wire (0.53 g, 76 mg- atom) cut in small pieces. After refluxing under argon for 6 h the reaction was quenched with 5 ml of methanol and ammolonia was allowed to boil off overnight. A suspension of the residue in 100 ml of water was extracted twice with 100 ml portions of ethyl acetate and the combined organic extracts were washed with brine and dried over magnesium sulfate. Following solvent removal under reduced pressure the residue was flash chromatographed on silica gel using 1% ethyl acetate/hexane as eluent and then recrystallized from abs. ethanol to give fluffy white crystals (884.1 mg, 3.269 mmole, 72%), m.p. 72-73°C, homogeneous to TLC. Example 5 - Synthesis of Estra-5(10) .16-dien-3-one (3) . Estra-2,5(10) ,16-triene-3-methyl ether (2) (646.3 mg, 2.390 mmole), dissolved in 50 ml of acetone was hydrolyzed for 6 h at room temperature using oxalic acid dihydrate (0.84 g, 6.7 mmole) . The reaction mixture was quenched with 25 ml of saturated sodium bicarbonate and then extracted twice with 25 ml portions of ethyl acetate. The combined organic extracts were washed twice with 25 ml of brine, dried over magnesium sulfate, filtered, and concentrated under reduced pressure. The residue was recrystallized from hexane to give product (462.5 mg, 1.804 mmole, 75%), m.p. 112-116°C. Example 6 - Synthesis of Estra-5(10) .16-dien-3-ols (4) . Estra-5(10) ,16-dien-3-one (3) (301.1 mg, 1.174 mmole) , in 6 mL of anhydrous ether was reduced for 1 h at room temperature using lithium aluminum hydride (50.0 mg, 1.32 mmole). After quenching with sodium sulfate decahydrate (2.00 g) for 10 min. the suspension was filtered through Celite and the residue washed with four 25 mL portions of ether. The combined filtrates were concentrated under reduced pressure and purified by flash chromatography (silica gel, 5% ethyl acetate/ hexanes eluent) with subsequent preparative TLC of mixed fractions. The more polar product could be recrystallized with considerable loss from aqueous ethanol to give 4.8 mg of solid. The less polar product was recrystallized from aqueous methanol to give white crystals (59.5 mg) , m.p. 98-100°C. Total yield was 64.3 mg (0.249 mmol, 21%). Example 7 - Synthesis of Estra-4.9.16-trien-3-one (5). Estra-5(10) ,16-dien-3-one (3) (0.38 g, 1.5 mmole), in pyridine (5.0 mL, 62 mmol) was cooled in an ice-salt bath to -13°C and pyridinium bromide perbromide (1.58 g, 4.94 mmole) was added in small portions so that T<-4°C. After swirling 1 min. phenol (0.25 g, 2.7 mmole) was added and reaction continued 24 h at room temperature. Ethyl acetate (50 ml) was added and the reaction mixture was washed with 25 ml of IN HCI, two 25 ml portions of saturated copper sulfate, 25 ml of 5% sodium hydroxide, and 25 ml of brine. After drying over magnesium sulfate, filtration, and concentration under reduced pressure the residue was taken up in 10 mL of abs. ethanol, granular zinc (0.33 g, 5.0 mg-atom) was added, and the mixture was refluxed 1/2 h. The supernatant was removed, the residue was washed with 10 mL of abs. ethanol, and the combined supematants were concentrated under reduced pressure. The resulting resin was flash chromatographed on silica gel using 15% ethyl acetate/ hexane as eluent. Appropriate fractions were pooled, concentrated, and then recrystallized from hexane to give solid product (117.5 mg, 0.4619 mmol, 31%) , m.p. 87-92°C. Example 8 - Synthesis of Estra-1.3.5(10) .16-tetraen-6- one-3-acetate (6) . Chromium trioxide (13.40 g, 0.1340 mol) was suspended in 200 mL of methylene chloride and then cooled to -10°C in an ice-salt bath. 3,5- Dimethylpyrazole (12.90 g, 0.1342 mol) was added and the mixturewas stirred 20 min. Estra-l,3,5(10) ,16-tetraen- 3-yl acetate (4.00 g, 13.5 mmol) in a chilled solution of 20 mL of methylene chloride was added and the reaction stirred 2 h, during which time T<-8°C. The mixture was then filtered through 200 g of silica gel and the product was eluted with further methylene chloride. After combining and concentrating appropriate fractions the crude product was flash chromatographed on silica gel using 15% ethyl acetate/hexane as eluent. Pooling of appropriate fractions and concentration under reduced pressure yielded a white solid (0.92 g, 3.0 mmol, 22%), m.p. 87-103°C. Example 9 - Synthesis of Estra-1.3.5(10) .16-tetraen-3- ol-6-one (7) . Estra-1,3,5(10) ,16-tetraen-6-one-3-acetate (203.1 mg, 0.6543 mmol) in 30 of methanol was saponified with 1.5 mL of 5% (w/w) sodium hydroxide for 40 min. The reaction mixture was concentrated under reduced pressure, taken up in 50 mL of water, neutralized with IN HCI, and extracted three times with 25 mL portions of methylene chloride. The combined organic extracts were washed with 50 mL of brine, dried over magnesium sulfate, filtered, and concentrated to give a white solid which was purified by recrystallization from benzene/hexane and preparative TLC to give white crystalline solid (52.8 mg, 0.197 mmol, 30%), m.p. 188- 191°C. Example 10 - Synthesis of Estra-1.3.5(10) .16-tetraen-6tt- ol-3-yl acetate (8) . Estra-1,3,5 (10) , 16-tetraen-6-one-3-yl-acetate (6) (421.4 mg, 1.358 mmol), suspended in 35 mL of 95% ethanol was reduced with sodium borohydride (98.8 mg, 2.61 mmol) for 100 min. at room temperature. After concentrating under reduced pressure the residue was suspended in 25 mL of water, neutralized with IN HC1, and extracted three times with 25 mL portions of methylene chloride. The combined organic extracts were washed with 25 mL of brine, dried over magnesium sulfate, filtered, and concentrated. The resulting white foam was flash chromatographed on silica gel using 25% ethyl acetate/hexane as eluent. Combining fractions and concentration gave a white solid (0.12 g, 0.38 mmol, 28%) , m.p. 209-212°C. Example 11 - Synthesis of Estra-1.3.5(10) .16-tetraene- 3.6-diol (9). To a suspension of lithium aluminum hydride (LAH, 95%, 46.9 mg, 1.17 mmol) in 5 mL of anhydrous THF was added estra-l,3,5(10) ,16-tetraen-6-one-3-yl-acetate (6) (422.9 mg, 1.360 mmol) in 5 mL of anhydrous THF dropwise, with stirring. The reaction was stirred 50 min., after which further LAH (46.5 mg, 1.16 mmol) was added and the reaction stirred 22 h. After refluxing 4 h TLC still showed starting material. The reaction was quenched with 0.5 mL of water + 0.5 mL of 20% (w/w) sulfuric acid and concentrated under reduced pressure. The residue was extracted four times with 10 mL portions of hot ethyl acetate and filtered through Celite. The combined filtrates were concentrated and purified twice by flash chromatography to give solid product (0.05 g, 0.2 mmol, 10%), m.p. 150-157°C. Exa ple 12 - Synthesis of Estra-1.3.5 (10) .7-tetraen-3-ol t lθ) . To a suspension of equilin (100.2 mg, 0.3733 mmol) in 2 mL of diethylene glycol were added hydrazine (59 μl>, 1.9 mmol) and potassium hydroxide (0.04 g, 0.7 mmol) . The mixture was stirred in an oil bath at 200-214°C for 2 h, after which the cooled reaction was diluted with 10 mL of water, neutralized with IN HC1, and extracted three times with 25 mL of ether. The combined organic extracts were washed with 10 mL of brine, dried over magnesium sulfate, filtered, concentrated, and purified by preparative TLC (silica gel, 15% ethyl acetate/hexane eluent) to give a yellow resin. Product was further purified by decolorizing with charcoal and recrystallization from aqueous ethanol to give tan crystals (13.2 mg, 51.9 μK, 14%), m.p. 130- 134°C. Example 13 - Synthesis of 20-Homoestra-l.3.5(10) . 6.8.17-hexaen-3-ol (11). A suspension of triphenylmethylphosphonium bromide (671.0 mg, 1.878 mg) and potassium t-butoxide (212.1 mg, 1.890 mmol) in 2.1 mL of anhydrous DMSO was heated in a 76-86°C bath under argon for 1 h, after which equilenin (100.1 mg, 0.3579 mmol) in 2.1 L of anh. DMSO was added and the green solution was stirred 1 h. After cooling 10 mL of ice-lN HC1 were added and the mixture was extracted with three 10 mL portions of ether. The combined organic extracts were washed with 10 mL of saturated sodium bicarbonate + 10 mL of brine, dried over magnesium sulfate, filtered through Celite, and concentrated under reduced pressure. The residual orange oil was purified by preparative TLC (silica gel, 25% ethyl acetate/hexane) to give product (75.5 mg, 0.286 mmol, 76%) homogeneous to TLC, m.p. 113-121°C. Example 14 - Synthesis of Estra-1.3.5(1Q) .6-tetraen-3-ol (17): Estra-l,3,5(10) ,6-tetraen-3-ol-17-one (91.1mg, 0.339 mmol), hydrazine (54 μh, 1.7 mmol), and potassium hydroxide (0.06 g) in 1.8 L of diethylene glycol were heated in a 200°C bath under argon for 2 h. After cooling to RT 10 L of water were added and the solution was acidified to pH«2 with IN HC1. The resulting suspension was extracted three times with 10 mL of ether and the combined organic extracts were washed with 10 mL of brine, dried over magnesium sulfate, filtered through Celite, and concentrated under reducedpressure. The crude solid was purified by preparative TLC (25% ethyl acetate/hexane on silica gel) to give product homogeneous to TLC (5.9 mg, 23 μmol, 7%). Example 15 - Svntesis of Estra-4.16-dien-3-ol (18): To estra-4,16-dien-3-one, (l) (87.2 mg, 0.340 mmol) in 1.7 mL of anh. ether was added lithium aluminum hydride (15.0 mg, 0.395 mmol) and the suspension was stirred 17 min. Reaction was then agitated 10 min. with 0.50 g of sodium sulfate decahydrate and filtered through Celite. The residue was washed with three 10 mL portions of ether and the combined filtrates were concentrated under reduced pressure. Preparative TLC (5% ethyl acetate/dichloro-methane on silica gel) gave crude product (50.0 mg) as a yellow resin. This could be rechromatographed until sufficiently pure. Example 16 - Estra-4.16-dien-3-one (9). This synthesis is depicted in Figure 11. 19-Nor- testosterone (XIX) is commercially available, e.g., from Chemical Dynamics Corp. It provides the starting material for 19-Nor-16-androstene derivatives D 19-Nor- testosterone (XIX) was converted into the acetate (Hartman, J.A. et al.. J. Am. Chem. Soc. (1956) 78:5662) with acetanhydride and pyridine (a) D A solution of this acetate (4.8 g, 15.17 mmol) in toluene (10 ml) was pyrolyzed (b) at 540° (200 Torr, slow N 2 -stream) in a glass tube packed with quartz pieces. Chromatography of the crude pyrolysate (3.1 g) on silica gel (150 g) with CH 2 Cl 2 gave 1.1 g (28%) of the homogenous oily ketone 9; +57.9° (C 1) ((27]: m.p. 71-73°). - IR. (CHC1 3 ) : 1660s, 1615m, 1585W, - 'H-NMR. (90 MHz): 0.84 (s, 3 H) ; 5.82 (m, 2 H) ; 5.87 (br. s, 1 H) . Example 17 - Esta-16-en-3-one (10) . This synthesis is depicted in Figure 11. 19-nor- testosterone was reduced to 19-nor-5a-Androstan-17 -ol-3 -one (XX) with lithium and ammonia (c) according to the method of Villotti, R. , et al. (J. Am. Chem. Soc. (1960) 8_2 . :5693). Androsta-5a,17-diol-3-one (XX) was converted into the acetate (Hartman, J.A. et al.. J. Am. Chem. Soc. (1956) 8. 5662) with acetanhydride and pyridine (a) . A solution of 17B-acetoxy-5a-Estran-3-one (8.0 g, 25.1 mmol) in octane/acetone 10:1 (22 ml) was pyrolyzed (b) at 550° (200 Torr, slow N 2 -stream) Chromatography of the crude product (5.4 g) on silica gel (600 g) with CH 2 Cl 2 and recrystallization of the homogenous fractions from PE gave 3.13 g (48.3%). of the pure ketone 10. M.p. 51-54°, [a]- +72.8° (C 1.0). -IR. (CHC1 3 ) : 1705s, 1585W, - -NMR. (90 MHz): 0.79 (s, 3 H) ; 5.71 (i, 1 H) ; 5.87 ( , 1 H) . Example 18 - Estra-16-en-3o:-ol (11) . This synthesis is depicted in Figure 11. L- Selectride (d, lithium tri(sec-butyl)hydridoborate, 4 ml of a 1 M solution in THF, 4 mmol) was added dropwise at 0° to a solution of ketone 10 (800 mg, 3.10 mmol) in dry ether (5 ml). After stirring for 1 h at 0°, water was added (10 ml) . The boranes were oxidized by adding 10% aq. NaOH-solution (5 ml) , followed by 30% aq. H 2 0 2 - solution (3 ml) and stirring for 3 h at RT. After workup (ether), the crude product (790 mg, Ca. 9:1 mixture of 11 and 12) was chromatographed on silica gel with CH 2 Cl 2 to give 700 mg (87%) of pure alcohol 11. M.p. 119-120°→123-124° (from PE) , [a] D +40.6° (C = 1.0). - IR. (CHC1 3 ) : 3640m, 3500 br., 1585w. - Η-NMR. (90 MHz) : 0.78 (s, 3 H) ; 4.09 (m, w, Λ « 8, 1 H) ; 5.71 (m, 1 H) , 5.87 ( , 1 H) . Example 19 - Estra-16-en-3B-ol (12) . This synthesis is depicted in Figure 11. A solution of the ketone 10 (800 mg, 3.10 mmol) in dry ether (5 ml) was added dropwise at RT. to a slurry of LiAlH 4 (38 mg, 1 mmol) in ether (3 ml) (e) . After 1 h, the mixture was hydrolyzed with 10% aq. H 2 S0 4 . After workup (ether), the crude product (802 mg, 9:l-mixture of 12 and 11) was chromatographed on silica gel with CH 2 C1 2 . A small fraction of 11 (70 mg) was eluted first, followed by the main fraction of 12 (705 mg, 87%) . M.p. 113-115°, [a]- +36.3° (C = 1.0). - IR. (CHC1 3 ) : 3640m, 3500 br., 1585w. - Η-NMR. (90 MHz) : 0.78 (s, 3 H) ; 3.60 (Jϊl, W. Λ « (m, 20, 1 H) ; 5.71 (m, 1 H) , 5.87 (m, 1 H) . Example 20 - Electrophvsiology of Estrene Stimulation of the Human VNO and Olfactory Epithelium. A non-invasive method has been employed to record local electrical potentials from the human vomeronasal organ (VNO) and from the olfactory epithelium (OE) . Localized gaseous stimulation was applied to both nasal structures at different instances using specially designed catheter/electrodes connected to a multichannel drug delivery system. The local response of the VNO and the OE showed a correlation with the concentration of the ligand stimulus. The study was performed on ten clinically normal (screened) volunteers - 2 males and 8 females, ranging in age from 18 to 85 years. The studies were conducted without general or local anesthetics. The catheter/electrodes were designed to deliver a localized stimulus and simultaneously record the response. In the case of VNO recording, the right nasal fosa of the subject was explored using a nasoscope (nasal specula) and the vomeronasal opening was localized close to the intersection of the anterior edge of the vomer and the nasal floor. The catheter/electrode was gently driven through the VNO- opening and the electrode tip placed in the organ's lumen at 1 to 3 mm from the opening. The nasoscope was then removed. In the case of the OE, recording the procedure was similar except the positioning of the catheter/electrode was gently placed deep in the lateral part of the medial nasal duct, reaching the olfactory mucosa. Localized gaseous stimulation was done through the catheter/electrode. A constant stream of clean, nonodorous, humidified air at room temperature was continuously passed through a channel of the stimulating system. The stimulating ligand substances were diluted in propylene glycol, mixed with the humidified air, and puffed for from 1 to 2 seconds through the catheter/electrode. It is estimated that this administration provides about 25 pg of steroid-ligand to the nasal cavity. The results of this study are presented in Figure 2. The response is measured in millivolt-seconds ( V x s) . 1,3,5(10) ,16-Estratetraen-3-ol elicits a significantly stronger VNO response in males than do the other compounds tested (Fig.2A) . 1,3,5(10)-Estratrien- 3, 16α,176-triol also elicits a strong VNO response. Furthermore, the VNO response to these two Estrenes is sexually dimorphic -approximately four times as strong in males as it is in females (Fig. 2B) . In contrast, the OE response in both males and females is low compared to a strong odorant such as clove (Fig. 2C) . Example 21 - Measurement of the Change in Receptor Potential of the Neuroepithelium of the VNO in Response to Various steroids. The change in receptor potential in response to seven different ligands was measured in 40 female (Fig. 3A) and 40 male (Fig. 3B) subjects. Each subject was administered 60 pg of each of seven substances as indicated in the Figures. The substances were administered, each separately for 1 second, using the procedure described in Example 20. The change in potential of the neuroepithelium of VNO was recorded over time and the integral of the change in potential for each of the forty subjects was averaged. The results are shown in the figure. Comparison of Figures 3A and 3B show that each steroid is sexually dimorphic in its activity and that some ligand substances are stronger in males while others are stronger in females. Example 22 - Measurement of Autonomic Responses to Estrene Stimulation of the VNO. Various autonomic parameters were monitored while 1,3,5(10) , 16-Estratetraen-3-yl-acetate was administered to 40 male subjects using the procedure described in Example 20. Propylene glycol was also administered as a control. The ligand was administered as a 1 second pulse. The change in autonomic function was first noted within 2 seconds and lasted for up to 45 seconds. As shown in Figure 4, when compared to a propylene glycol control, the Estrene, induced a significant change in the integrated receptor potential in the VNO (4A) , galvanic skin response (4B) , and skin temperature (4C) . Example 23 - Comparison of the Change in Receptor Potential Induced by Two Estrene Steroids 60 picograms of each steroid and of a propylene glycol control were administered to a male subject as described in Example 21. As shown in Figure 5, 1, 3 ,5 (10) ,16-Estratetraen-3-ol methyl ether induced a greater change in receptor potential than did 1,3,5(10) ,16- Estratetraen-3-yl acetate. Example 24 - Psychophvsiological Effect of Estrene Stimulation of the VNO. The psychophysiological effect of Estrene stimulation of the VNO is measured by the coordinate administration of pheromone and questionnaire evaluation of the subject before and after administration. The questionnaire includes a panel of adjectives used as part of the standard Derogatis Sexual Inventory evaluation. 40 subjects, all in good health, are randomly assigned - 20 exposed to placebo and 20 exposed to about 20 picograms of 1,3,5(10) ,16-Estratetraen-3-ol, administered as described in Example 3, supra. Subjects are given a 70 item questionnaire evaluating feeling states immediately before and 30 minutes after administration of either placebo or experimental substance. The 70 adjectives of the questionnaire are randomly administered and subsequently clustered for evaluation based on their relevance to each mood, feeling, or character trait. Example 25 - Electrophysiological Studies The following electrophysiological studies were performed in 60 clinically normal human volunteers of both sexes (30 male and 30 female) whose ages ranged from 20 to 45 years. No anesthetics were used, and female subjects were excluded if pregnant. The stimulation and recording system consists of a "multifunctional miniprobe" described elsewhere (Monti-Bloch, L. and Grosser, B.l. (1991) "Effect of putative pheromones on the electrical activity of the human vomeronasal organ and olfactory epithelium," J. Steroid Biochem. Molec. Biol. 29:573-582.). The recording electrode is a 0.3 mm silver ball attached to a small (0.1 mm) silver wire insulated with Teflon© the surface of the electrode is first treated to produce a silver chloride interface, and is then covered with gelatin It is positioned within a small caliber Teflon© catheter (dia = 5 mm) such that the tip of the electrode protrudes approximately 2 mm. The Teflon© catheter is 10 cm in length and constitutes the terminal extension for a multichannel delivery system which delivers a continuous air stream carrying discreet pulses of chemosensory stimuli. The air stream first passes into a small chamber and is bubbled through a solution containing either a vomeropherin or an olfactant in a diluent or the diluent alone. A solenoid is used to rapidly redirect the air stream from the chamber to a route which bypasses the chamber. This creates a discreet pulse of stimulant in the air stream. A second, outer Teflon® tube with a diameter of 2 mm surrounds the catheter-electrode assemblage, and its central end is connected to an aspirator that provides continuous suction of 3ml/s. This concentric arrangement of the outer suction tube allows the emitted chemosensory stimuli to be localized to an area we call a "minifield" (approx. dia = 1 mm) , and it avoids diffusion of substances either to the area outside the intended stimulation site or into the respiratory system. The entire stimulating and recording assemblage may be positioned either on the neurosensory epithelium within the VNO, or on the surface of the olfactory or respiratory epithelium. Electro-vomeronasogram (EVG) : Recordings are carried out in a quiet room with the subject supine; the multi¬ functional miniprobe is initially stabilized within the nasal cavity using a nasal retractor placed in the vestibule. Reference and ground electrodes consist of silver discs (8 mm) , both of which are positioned on the glabella. The entrance to the VNO, or vomeronasal pit, is identified by first dilating the nasal aperture and vestibule. A 6x magnifying binocular loupe with halogen illumination is then used to introduce the tip of the Teflon® catheter and recording electrode assemblage into the VNO opening where it is stabilized at an approximate depth of 1 mm within the vomeronasal passage. Optimal placement of the recording electrode is signaled after testing for an adequate depolarization in response to a test substance. Electrical signals from the recording electrode are fed to a DC amplifier after which they are digitized, computer monitored, and stored. The peak-to- peak amplitude of the signals is measured, and the area under the depolarization wave is integrated, while continuously monitoring the signal both on the computer screen and on a digital oscilloscope. Artifacts produced by respiratory movements are deleted by training the subjects to practice mouth breathing with velopharyngeal closure. Chemosensory Stimulants: Olfactory test substances are cineole, and 1-carvone; vomeropherins are A, B, C, E and F. (Vomeropherins were supplied by Pherin Corporation, Menlo Park, California) , Samples of vomeropherins In concentration of 25-800 fmoles are delivered in the continuous air stream for durations from 300 milliseconds to 1 second. Usually, intervals of 3 to 5 minutes separated each series of short test pulses. All components of the lines carrying the test stimuli are made of Teflon®, glass or stainless steel and are carefully cleaned and sterilized before each use. Electro-olfactgram (EOG) : Olfactory recordings employed the same stimulating and recording multifunctional miniprobe as that used for the VNO. The tip was slowly introduced until the recording electrode touched the olfactory mucosa. Adequate placement was signaled by a depolarization in response to a pulse of the odorant test substance. Cortical evoked activity was induced by VNO stimulation with vomeropherins, and olfactory stimulation with odorants delivered in 300 ms air pulses. It was recorded using standard electroencephalographic (EEG) electrodes placed at positions Cz-Al and Tz-Al of the international 10120 system; the ground electrode was placed on the mastoid process. Electrodermal activity (EDA) was recorded using standard 8 mm silver electrodes in contact with palmar skin of the medial and ring fingers respectively, through a conductive gel interface. Skin temperature (ST) was recorded by a small (1.0 mm) thermistor probe placed in the right ear lobe. Peripheral arterial pulse (PAP) was monitored with a plethysmograph attached to the tip of the index finger. Respiratory frequency (RF) was measured with an adjustable strain gauge placed around the lower thorax. All electrical signals were DC amplified, digitized (MP-100, Biopac Systems) and continuously monitored utilizing a computer. Statistical Analysis: EVGs or EOGS, peak-to-peak changes and frequency changes of other parameters were measured and statistically analyzed. The significance of the results was determined by either using paired t- tests or analysis of variance (ANOVA) . Effect of Vomeropherins on the EVG: Each of the vomeropherins was found to produce a sexually dimorphic receptor potential (Fig. 6A-B) . Recordings of the EVG were performed on 30 men and 30 women (ages 20 to 45) . Vomeropherins were diluted and applied as 1 second pulses to the VNO with b minute intervals between pulses when questioned, the subjects were not able to "smell" or otherwise consciously detect any of the vomeropherins. This finding is in agreement with results previously reported (Monti-Bloch, L. and Grosser, B.l. (1991) "Effect of putative pheromones on the electrical activity of the human vomeronasal organ and olfactory epithelium," J. Steroid Biochem. Molec. Biol. 2£ 573-582.) which indicated that neither olfactory nor vomeropherin test stimuli delivered to the VNO elicit a perceptible sensation at the delivered concentration. Fig. 6A shows the average response of male subjects (ages 20 to 38) to the diluent, and to equimolar quantities (100 fmoles) of five vomeropherins (A, B, C, D , and F) , and to E, a stereoisomer of F. The profile of the response to each of the substances was similar in all subjects regardless of age, and no significant differences were revealed either by t-tests or by analysis of variance. For example, A, C and D produced significant effects (M 15 = 11.4 mV, SD = 3.6 mV; M 76 = 6.4 mV, SD 2.5 mV, and K M = 15.1 mV, SD = 4.9mV; p<0.01), that were consistent in all individual cases. Other vomeropherins depolarized the VNO-receptors to a much lesser extent, but with consistent mean response amplitudes from individual to individual. Vomeropherins active in male subjects produced larger responses than the diluent (p<0.001) . B, F and similar concentrations of olfactants induced significantly reduced responses in the male VNO (Fig. 6A and Fig. 7) . A similar experimental protocol was followed with the 30 female subjects (ages 20 - 45) . Among the vomeropherins, F (100 fmoles) produced the most significant differences within the group (Fig. 6B) . Here, A induced a small effect that was significantly different from F (p<0.01). In both populations of subjects, active vomeropherins induced receptor responses having large standard deviations (Fig. 6) . When the frequency distribution of the effects of A and F was studied in males and females respectively, we found a bimodal distribution. The significance of this observation is being studied at this point. E, a stereoisomer of F, does not stimulate the VNO in female subjects while F does (FIG 6B) . This is a demonstration of the specificity of VNO recognition of vomeropherins. In this regard it is interesting to note that while F is a superior vomeropherin, E generates a stronger olfactory effect than does F (Fig. 6B and Fig. 7) . Effects of Vomeropherins on the EOG: The sum ated receptor potential from the olfactory epithelium (OE) was recorded in 20 subjects: 10 males and 10 females. In contrast to the sensitivity of the VNO to vomeropherins, the OE is less sensitive to these substances. This is true for both males and females (Fig.7A) . The mean receptor potential amplitude ranged from 2.3 mV to 0.78 V. In this study, B was the only vomeropherin having significant effect in the OE (p<0.02). Of the subjects questioned about odorant sensations following each stimulus presentation, 16 reported no olfactory sensation, while three males and one female described B as an unpleasant odor. This finding reveals that at the concentrations used in our study, most vomeropherins are not effective stimulants of the olfactory receptors, but do have a clear effect on vomeronasal receptors. Effects of Olfactants on the EVG and EOG: In contrast to vomeropherins, the olfactants 1-carvone and cineole produce only a minor local response in the VNO (Fig. 7B) . This was true for both men and women. As expected, these olfactants produced a strong response in both men and women (p<0.01) when locally applied to the OE (Fig. 7A) . The diluent depolarized olfactory receptors to a lesser extent than cineole or 1-carvone (p<0.01), and it did not produce an olfactory sensation. Reflex Effects of Vomeropherins: Studies were conducted to determine the central nervous system (CNS) reflex responses to vomeropherin stimulation of the VNO. The sexually dimorphic local responses induced by vomeropherins (Fig. 6A and B) were mirrored in the autonomic response of male & female subjects. In male subjects (Fig. 6C) , A and C decreased skin resistance (electrodermal acuity EDA) (p<0.01, n = 30) . In female subjects. (Fig. 6B) , F and B produced greater decrease in EDA than A or C (p<0.01, n = 30). Vomeropherins A and C induced a significant increase in skin temperature (ST) (Fig. 6G) in 30 male subjects (p<0.01); however D induced significant temperature decrease (p<0.01). In 30 female subjects (Fig. 6H) B and F evoked a significant increase in skin temperature (ST) (p<0.01) compared to A and C. In female subjects vomeropherins produced changes in EDA and ST with a greater standard deviation than in males. Cortical activity was recorded from Cz and Tz in male and female subjects during application to the VNO of air pulses (300 ms to 1 sec) containing 200 fmoles of vomeropherin (Fig. 6G and H) . In males (Fig. 6E) A, C and D significantly increased alpha cortical activity with a latency of 270-380 ms. D and A evoked the strongest effect (p< 0.01) . Synchronization of the EEG was sustained for 1.5 to 2.7 minutes after application of a single pulse of active substance. In females (Fig. 6F) , a single pulse (200 fmoles) of B or F applied to the VNO increased alpha cortical independent of the response of olfactory receptors. We found characteristic specificities in the response of the human VNO and the olfactory epithelium which suggests that they are independent functional systems with separate connections to the CNS (Brookover, C. (1914) The nervus terminalis in adult man. J. Comp. Neurol.2 :131-135.) There is also preliminary evidence that the EVG is not associated with trige inal nociceptor endings since application of a local anesthetic (2% lidocaine) to the respiratory epithelium of the nasal septum neither blocks nor diminishes the EVG (Monti-Bloch, L. and Grosser, B.l. (1991) "Effect of putative pheromones on the electrical activity of the human vomeronasal organ and olfactory epithelium," J. Steroid Biochem. Molec. Biol. 3_9:573-582.) , also, subjects failed to report sensations of pain as a consequence of any of the stimulation procedures. VNO receptors are clearly more sensitive to vomeropherins than to any of the olfactants tested; the opposite is true for olfactory receptors. While the OE may have receptor sites for some vomeropherins, the response specificity of the VNO is clearly different. Sexual differences were noted in the specificities and effects of two groups of vomeropherins. A, C and D; and B and F. This suggests a possible receptor-related sexual dimorphism. The findings suggest the activation of components of the autonomic nervous system In the adult human by vomeropherin stimulation of the VNO. Furthermore, the results suggest that stimulation of the VNO with vomeropherins produces synchronization of the EEG (Fig. 6G and H) Thus, the evidence herein indicates that the vomeronasal system responds to a variety of chemosensory stimuli, and that some are able to induce reflex autonomic activity.

Claims: CLAIMS We claim: 1. A pharmaceutical composition suitable for nasal administration in a human subject, said composition comprising an Estrene steroid and, a pharmaceutically acceptable carrier, wherein said Estrene steroid has the formula: wherein R, is selected from the group consisting essentially of one or two hydrogen atoms, methyl, methylene, and one or two halo atoms; R 2 is absent or is selected from the group consisting essentially of hydrogen and methyl; R 3 is selected from the group consisting essentially of oxo, hydroxy, lower alkoxy, lower acyloxy, benzoyl, cypionyl, glucuronide and sulfonyl; R 4 is selected from the group consisting essentially of hydrogen, hydroxy, lower alkoxy, lower acyloxy and halo; R 5 is absent or is selected from the group consisting essentially of hydrogen, hydroxy, lower alkoxy and lower acyloxy; Rg is a hydrogen or a halo; and "a" represents optional aromatic unsaturation of ring A of said steroid, or "b", "c", and "d" are each optional double bonds; and "e", "f", "g", "h", "i" and "j" are each optional double bonds. 2. A composition according to Claim 1 wherein "a" is present and "g", "h" or "i" are optional double bonds. 3. A composition according to Claim 2 wherein "h" and "i" are both double bonds. 4. A composition according to Claim 1 wherein "b" is a double bond. 5. A composition according to Claim 1 wherein "j" is a double bond. 6. A composition according to Claim 1 wherein "c" is a double bond. 7. A composition according to Claim 1 wherein "c" and "d" are double bonds. 8. A composition according to Claim 1 wherein R 2 is methyl and "e" is a double bond. 9. A composition according to Claim 1 wherein said steroid is selected from the group consisting of Estra-4,16-dien-3-one;Estra-l,3,5(10) ,16-tetraene-3-ol; Estra-4,16-dien-3o-ol; Estra-4,9(10,16-triene-3-one; Estra-1,3,5(10) ,16-tetraen-3-ol-6-one;3-Methoxyl-estra- 2,5(10) ,16-triene; Estra-5(10) ,16-dien-3α-ol; and Estra- 1,3,5(10) ,16-tetraen-3,6α-diol. 10. A composition according to Claim 1 wherein R 2 is methyl. 11. A composition according to Claim 10 wherein said steroid is selected from the group consisting essentially of Estra-1,3,5(10)-trien-3-ol; Estra-l,3,5(10) ,6-tetraen-3-ol; and Estra-l,3,5(10) ,7- tetraen-3-ol. 12. A composition according to Claim 1 wherein Rj is methylene. 13. A composition according to Claim 12 wherein said steroid is 17-Methylene-estra- 1,3,5(10) ,6,8(9)-hexaen-3-ol. 14. A composition according to Claim 1 wherein R, is methylene or a single hydrogen and R 2 is methyl. 15. A composition according to Claim 1 wherein "f" is a double bond and R 2 is methyl. 16. The pharmaceutical composition of Claim 1 wherein the Estrene steroid is selected from the group consisting of 1,3,5(10) ,16-Estratetraen-3-ol, 1,3,5(10) , 16-Estratetraen-3-ol methyl ether, 1, 3 , 5 (10) , 16-Estratetraen-3-yl acetate, and 1,3,5(10) ,16-Estratetraen-3-yl propionate. 17. The pharmaceutical composition according to any of Claims 1 through 16 wherein said at least one Estrene steroid is dissolved in said carrier. 18. The pharmaceutical composition of any of Claims 1 through 16 wherein said composition is in a liquid form. 19. The pharmaceutical composition of any of Claims 1 through 16 wherein said composition further contains a pharmaceutically acceptable ointment base. 20. The pharmaceutical composition of any of Claims 1 through 16 which contains no more than one Estrene steroid. 21. The pharmaceutical composition of any of Claims 1 through 16 which contains more than one Estrene steroid. 22. The pharmaceutical composition of any of Claims 1 through 16 additionally comprising a 16- Androstene steroid. 23. A method of altering a hypothalamic function of an individual, said method comprising: providing a ligand for a chemoreceptor displayed on the surface of nasal neuroepithelial cell of said individual wherein said cell is a part of tissue other than olfactory epithelia; and, administering said ligand within a nasal passage of said individual such that said ligand binds specifically to said receptor and results in an alteration of hypothalamic function of said individual. 24. Amethod of altering an autonomic function of an individual, said method comprising: providing a ligand for a chemoreceptor of a nasal neuroepithelial cell of said individual wherein said- cell is a part of tissue other than olfactory epithelia; and, administering said ligand within a nasal passage of said individual such that said ligand binds specifically to said receptor and results in an alteration of hypothalamic function of said individual. 25. The method of claim 24 wherein said neuroepithelial cell is located within a vomeronasal organ of said individual. 26. The method of claim 25 wherein said ligand is an Estrene steroid. 27. The method of claim 26 wherein said ligand is an Estrene steroid which has the formula: wherein Rj is selected from the group consisting essentially of one or two hydrogen atoms, methyl, methylene, and one or two halo atoms; R 2 is absent or is selected from the group consisting essentially of hydrogen and methyl; R 3 is selected from the group consisting essentially of oxo, hydroxy, lower alkoxy, lower acyloxy, benzoyl, cypionyl, glucuronide and sulfonyl; R 4 is selected from the group consisting essentially of hydrogen, hydroxy, lower alkoxy, lower acyloxy and halo; R 5 is absent or is selected from the group consisting essentially of hydrogen, hydroxy, lower alkoxy and lower acyloxy; 1^ is a hydrogen or a halo; and "a" represents optional aromatic unsaturation of ring A of said steroid, or "b", "c", and "d" are each optional double bonds; and "e", "f", "g", "h", "i" and "j" are each optional double bonds. 28. Amethod according to Claim 27 wherein "a" is present and "g", "h" or "i" are optional double bonds. 29. Amethod according to Claim 28 wherein "h" and "i" are both double bonds. 30. A method according to Claim 27 wherein "b" is a double bond. 31. A method according to Claim 27 wherein "j" is a double bond. 32. A method according to Claim 27 wherein "c" is a double bond. 33. A method according to Claim 27 wherein "c" and "d" are double bonds. 34. A method according to Claim 27 wherein R 2 is methyl and "e" is a double bond. 35. A method according to Claim 27 wherein said steroid is selected from the group consisting of Estra-4,16-dien-3-one;Estra-l,3,5(10) ,16-tetraene-3-ol; Estra-4,16-dien-3α-ol; Estra-4,9(10,16-triene-3-one; Estra-1,3,5(10) ,16-tetraen-3-ol-6-one;3-Methoxyl-estra- 2,5(10) ,16-triene; Estra-5(10) ,16-dien-3α-ol; and Estra- 1,3,5(10) ,16-tetraen-3,6α-diol. 36. A method according to Claim 27 wherein R 2 is methyl. 37. A method according to Claim 36 wherein said steroid is selected from the group consisting essentially of Estra-l,3,5(10)-trien-3-ol; Estra- 1,3,5(10) ,6-tetraen-3-ol; and Estra-1,3,5(10) ,7-tetraen- 3-ol. 38. A method according to Claim 27 wherein R t is methylene. 39. A method according to Claim 38 wherein said steroid is 17-Methylene-estra-l,3,5(10) ,6,8(9)- hexaen-3-ol. 40. A method according to Claim 27 wherein R y is methylene or a single hydrogen and R 2 is methyl. 41. A method according to Claim 27 wherein "f" is a double bond and R 2 is methyl. 42. The method of claim 41 wherein at least one Estrene steroid is selected from the group consisting of 1, 3 , 5 (10) , 16-Estratetraen-3- ol,l,3,S(10) ,16-Estratetraen-3-ol methyl ether, 1,3,5(10) ,16-Estratetraen-3-yl acetate, and 1,3,5(10), 16-Estratetraen-3-yl propionate. 43. The method of claim 42 wherein the Estrene steroid is 1,3,5(10), 16-Estratetraen-3-ol. 44. The method of any of Claims 27 through 43 wherein the amount of said ligand that is administered is at least about 100 picograms. 45. The method of any of Claims 27 through 43 wherein the amount of said ligand that is administered is at least about l nanogra s, but no more than about 10 micrograms. 46. The method of any of Claims 27 through 43 wherein the amount of said ligand that is administered is at least about 10 nanograms, but no more than about 1 microgram. 47. The method of any of Claims 27 through 43 further comprising preparing a pharmaceutical composition of said ligand dissolved in a pharmaceutically acceptable carrier. 48. The method of any of Claims 27 through 43 wherein said pharmaceutical composition is an ointment. 49. The method of any of Claims 27 through 43 wherein said pharmaceutical composition is liquid. 50. The method of any of Claims 27 through 43 wherein the administration is by aerosol. 51. The method of any of Claims 27 through 43 wherein more than one Estrene steroid is administered. 52. The method of any of Claims 27 through 43 further comprising nasally co-administering to said individual a 16-Androstene steroid. 53. The method of any of Claims 27 through 43 wherein said individual is male. 54. The method of claim 27 wherein said function is the alleviation of symptoms of psychoses, depression and anxiety.

Inventors: ADAMS NATHAN WILLIAM, BERLINER DAVID L, JENNINGS-WHITE CLIVE L, ADAMS, NATHAN, WILLIAM, BERLINER, DAVID, L., JENNINGS-WHITE, CLIVE, L.
Applicants: PHERIN CORP, PHERIN CORPORATION
Application year: 1993
Main classification: A61K31
IPC codes: A61K31, A61P25, C07J1