US 2005O250772A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2005/0250772 A1 Shelby et al. (43) Pub. Date: Nov. 10, 2005

(54) METHODS FOR INDUCING ANTI-ANXIETY (60) Provisional application No. 60/587,167, filed on Jul. AND CALMING EFFECTS IN ANIMALS AND 12, 2004. Provisional application No. 60/196.829, HUMANS filed on Apr. 13, 2000. (76) Inventors: Nancy J. Shelby, Park City, UT (US); Publication Classification Mitchell T. Godfrey, Townsend, MT (US); Mark J. Rosenfeld, Draper, UT (51) Int. Cl." ...... A61K 31/538; A61K 31/423 (US) (52) U.S. Cl...... 514/230.5; 514/375 Correspondence Address: (57) ABSTRACT PATE PERCE & BARD Phenolic compounds with a phenolic molecule to which are 215 SOUTH STATE STREET, SUITE 550 covalently linked an oxygen-containing group, a nitrogen or PARKSIDE TOWER another oxygen containing group, and a C-C alkoxy group, SALT LAKE CITY, UT 84111 (US) or their precursor compounds, obtainable from monocoty ledonous , or by chemical Synthesis, have been found (21) Appl. No.: 11/178,998 to calm and/or reduce anxiety and related behaviors and (22) Filed: Jul. 11, 2005 States in humans and animals. Additional chemical com pounds of the present invention may include benzoxazi Related U.S. Application Data noids-cyclic hydroxyamic acids, lactams, and corresponding glucosides, which may serve as precursors to phenolic (63) Continuation-in-part of application No. 10/845,388, compounds. The phenolic compounds and precursors of filed on May 13, 2004. phenolic compounds of the present invention, at concentra Continuation-in-part of application No. 10/718,232, tions Suitable for human and animal therapeutic use, may be filed on Nov. 20, 2003, which is a continuation-in-part obtained from monocotyledonous plants Such as corn in of application No. 09/834,592, filed on Apr. 13, 2001, their early growth states which are timely harvested for now Pat. No. 6,667,308. optimum yield. Patent Application Publication Nov. 10, 2005 Sheet 1 of 4 US 2005/0250772 A1

Preservation of immune response to DNFB (ear swelling) in stressed Balb/c mice exposed to novel compounds of the present invention

U

t E E

F.G. 1 Patent Application Publication Nov. 10, 2005 Sheet 2 of 4 US 2005/0250772 A1

Immune response (earswelling) to DNFB in stressed Bab/c mice exposed to novel compounds of the present invention

FIG. 2 Patent Application Publication Nov. 10, 2005 Sheet 3 of 4 US 2005/0250772 A1

Immune response (earswelling) to DNFB in stressed calves exposed to novel compounds of the present invention

U O 8 9 0.6 E E O. 4

FIG. 3 Patent Application Publication Nov. 10, 2005 Sheet 4 of 4 US 2005/0250772 A1

Immune response (neck swelling) to DNFB in stressed caves exposed to novel compounds of the present invention

FIG. 4 US 2005/0250772 A1 Nov. 10, 2005

METHODS FOR INDUCING ANT-ANXIETY AND when treated with antidepressants, many do not respond to CALMING EFFECTS IN ANIMALS AND HUMANS the first antidepressant prescribed. Such individuals or ani mals may eventually benefit from a different antidepressant RELATED APPLICATIONS or, if Sufficiently efficacious, a combination of Suitable antidepressants. 0001) This application claims the benefit of U.S. Patent Application Ser. No. 60/587,167, filed Jul. 12, 2004, and 0007 Sexual dysfunction is another pervasive disorder. entitled “METHODS FOR INDUCING ANTI-ANXIETY In the overall population, it is believed that forty-three AND CALMING EFFECTS IN ANIMALS AND percent (43%) of women and thirty-one percent (31%) of HUMANS,” and is a continuation-in-part of U.S. patent men between the ages of 18 and 59 experience Sexual application Ser. No. 10/845,388, filed May 13, 2004, and dysfunction and Some repeatedly. AS will be appreciated, entitled “METHODS FOR AUGMENTING IMMUNE Sexual dysfunction may include, for example, a lack of DEFENSES CONTEMPLATING THE ADMINISTRA interest in SeX, problems with arousal, not enjoying Sex, and TION OF PHENOLIC AND INDOLEAMINE-LIKE COM anxiety associated with Sexual performance. Indeed, feeling POUNDS FOR USE IN ANIMALS AND HUMANS and good in general has been found to have a significant impact U.S. patent application Ser. No. 10/718,232, filed Nov. 20, on Sexual function. In this regard, those people who are 2003, and entitled “NOVEL COMPOUNDS FOR USE IN unhappy or depressed may be more likely to experience WEIGHT LOSS AND APPETITE SUPPRESSION 1N difficulties associated with Sexual dysfunction. Arousal HUMANS,” which claim the benefit of U.S. patent appli problems affect over 20 million American males (i.e., about cation Ser. No. 09/834,592, filed Apr. 13, 2001, and entitled 2 in 10 adult men), with such difficulties often associated “NOVEL COMPOUNDS FOR USE AS ANTIDEPRES with or accompanied by Some Sort of depression or mood SANTS, APHRODISIACS, AND ADJUNCTIVE THERA disorder. Meanwhile, prescription antidepressants have been PIES 1N HUMANS, now issued as U.S. Pat. No. 6,667, found to actually exacerbate the Situation, Since a frequent 308, which claims the benefit of U.S. provisional application Side effect of their use is Sexual dysfunction. In fact, Sexual Ser. No. 60/196.829, filed Apr. 13, 2000, and entitled response may actually diminish in up to Seventy-five percent “ANTIDEPRESSANT, APHRODISIAC, WEIGHT-LOSS (75%) of prescription antidepressant users for using the AIDE, THERAPY FOR QUITTING NICOTINE OR Same. To this end, there is a need for forms of treatment that ADDICTIVE DRUGS AND TREATMENT OF BETTER reduce depression or otherwise better mood with an accom ING REPRODUCTION 1N HUMANS, all of which are panying enhancement of Sexual response, arousal or desire hereby incorporated herein by reference. or, in the alternative, do not have a negative impact on a perSon's or animal's Sexual function. BACKGROUND 0008 Prior work by those skilled in the art in connection with the compounds of the present invention have been 0002) 1. Field of the Invention primarily focused on 6-methoxy-2,3-benzoxazolinone 0003. This invention relates to compositions for calming (6-MBOA). For example, the role of 6-MBOA in strength and reducing anxiety in humans and animals and, more ening the resistance of monocotyledonous plants against a particularly, to novel compositions of phenolic and wide range of insect pests has been much Studied. It has been indoleamine-like compounds obtainable in concentrations further proven that 6-MBOA and its chemical precursors and amounts Suitable for animal and human use derived, may also have allelopathic properties that may inhibit root isolated, and/or extracted from certain botanicals or from and shoot growth in competing Species. Moreover, it has chemical Synthesis, together with methods for using, pro been found that 6-MBOA may have antimicrobial proper ducing, and harvesting the same for use in calming and/or ties, wherein 6-MBOA appears constitutively during early reducing anxiety in humans and animals Suffering from Stages of growth, localized in those tissues most exposed to anxiety disorders and related behaviors or during periods of microbial and insect attack. StreSS, illness, or injury. 0009. It had been long suspected that compounds in 0004 2. The Background Art plants affect the Seasonal reproductive output of wild 0005. An estimated 35-40 million living Americans will rodents. In 1981, 6-MBOA became the first naturally occur Suffer major depressive episodes, and many more will expe ring compound in a that was verified as impacting rience lesser bouts. Of the approximately 17.5 million Seasonal reproductive cycling. AS readily known, 6-MBOA Americans with ongoing depressions, about 9.2 million are may be found in varying concentrations in at a clinically debilitating level. Clinical depression is char ous plants. Since then, a Substantial body of work has acterized by a list of Symptoms that last over a long time accumulated on 6-MBOA as an initiator of seasonal breed Span. AS appreciated by those skilled in the art, depression ing and an effector of population size for many rodents and is a Serious problem that is generally initiated or caused by a few birds. Compounds related to and possibly co-occur outside StreSSors. AS Stresses escalate or persist, a chemical ring with 6-MBOA remain generally unexplored in this imbalance of the body usually results. regard. 0006 Clinical depression can be very debilitating both 0010) 6-MBOA may be passed from adult females to physically and mentally and even lead to death by means of offspring during gestation and lactation, which typically Suicide. Lost productivity and relationship problems may be results in increased growth and larger gonads in the recipient consequences of lesser depressions. Presently, antidepres young. Juveniles rely on the interaction of maternal photo Sant medications are the cornerstones of treating depression, period history and 6-MBOA to time the onset of growth and especially those that are at least moderately Severe. puberty. It has been found that adult animals that are fed a Although depressed individuals and animals tend to improve diet containing 6-MBOA produce more female progeny. US 2005/0250772 A1 Nov. 10, 2005

Similarly, when 6-MBOA is fed to pregnant females, 0015 The inventors of the present invention recognized gonadal development in the male offspring is usually that 6-MBOA and the indoleamine, melatonin, share a enhanced. Structural Similarity. However, melatonin exacerbates Symp 0011. In rodents, it has been found that the inhibitory toms of dysphoria in depressed people. 6-MBOA, as a effects of melatonin on growth and reproduction are blocked melatonin agonist, could prove contrary in this regard and partially by 6-MBOA (Gower et al., “Reproductive actually improve mood. Yet, the inventors are not aware of responses of male Microtus montanus to photo period, any prior art that has explored or Suggested the use of melatonin, and 6-MBOA”, Journal of Pineal Research, 8: 6-MBOA or related compounds as having psychotropic 297-312, 1990). As further shown by those skilled in the art, effects in humans, particularly with respect to depression or 6-MBOA may obstruct melatonin at the melatonin receptors bettering mood. An object of the present invention is to or act independently to check melatonin action (Sweat et al., develop therapies for depression and Sexual dysfunction “Uterotropic 6-methoxybenzoxazolinone is an adrenergic entailing use of compounds belonging to related chemical families, of which 6-MBOA is a member. Pursuant to this agonist and melatonin analog, Molecular and Cellular end, a further object is to develop methods for extracting, Endocrinology, 57:131-138, 1988). deriving, and/or isolating Said compounds from plant and 0012. The high melatonin levels induced by 6-MBOA animal Sources in amounts Suitable for human therapeutic may also cause desensitization of melatonin receptors (Daya Sc. et al., “Effect of 6-methoxy-2-benzoxazolinone on the 0016. In addition to the effects of stress on depression and activities of rat pineal N-acetyltransferase and hydroxyin mood disorders, as well as Sexual performance (i.e., desire, dole-O-methyltransferase and on melatonin production, arousal and performance), as outlined hereinabove, those ''Journal of Pineal Research, 8:57-66, 1990), but not for all skilled in the art may also appreciate the multiple effects of rodents (Anderson et al., “Effects of melatonin and 6-meth StreSS on the immune Systems and anxiety levels in animals oxybenzoxazolinone on photoperiodic control of testis size and humans. For example, animals may be at risk for in adult male golden hamsters,” Journal of Pineal Research, immune dysfunction related to StreSS which may manifest as 5:351-65, 1988). increased infections, morbidity, and mortality associated 0013 Further findings of those skilled in the art suggest with Shipping StreSS, Surgery, weaning, performance or train that 6-MBOA stimulates rather than inhibits melatonin bio ing, crowding, and/or mixing with unfamiliar animals. The Synthesis and does not prevent Stimulation of melatonin costs of Such immune impairment to the livestock and other synthesis by norepinephrine (Yuwiler et al., “Effects of food animal industries may be significant and is estimated at 6-methoxy-2-benzoxazolinone on the pineal melatoningen over S1 billion per year in the United States alone. erating system.J. Pharmacol. Exp. Ther. 233:45-50, 1985). Moreover, 6-MBOA may act at both the alpha-(C) and 0017 Stress in animals and humans may be due to beta-(B) adrenergic receptors (Daya et al., “Effect of 6-meth physical, chemical or electromagnetic challenge, illness, oxy-2-benzoxazolinone on the activities of rat pineal injury; and/or psychologic factors. Typically, these factors N-acetyltransferase and hydroxyindole-O-methyltransferase induce physiological adaption within the body of the Stressed animal or human. Adaption may manifest in a and on melatonin production, Journal of Pineal Research, plurality of responses. StreSS-induced changes in hormone 8:57-66, 1990), and may stimulate adenylcyclase (i.e., ade production may include higher levels of corticosterone, an nylate cyclase) activity in the pineal, hypothalmus and immunosuppressive hormone. StreSS and anxiety induced pituitary glands (Sweat et al., “Uterotropic 6-methoxy neuroendocrine responses may be associated with changes benzoxazolinone is an adrenergic agonist and melatonin in immune System function including alterations in levels of analog, Molecular and Cellular Endocrinology, 57:131 pro-inflammatory cytokines. These StreSS-induced responses 138, 1988). may, accordingly, result in an increased Susceptibility to 0.014 AS appreciated, certain responses to 6-MBOA, like Viral and bacterial infection, and a greater possibility of uterine hypertrophy, can be duplicated with estrogen, but complications associated with cytokine abnormalities and 6-MBOA is not an estrogenic compound (Gower, “Endo inflammatory processes. crine effects of the naturally occurring reproductive Stimu 0018 Humans may demonstrate immune abnormalities lant, 6-methoxy-benzoxazolinone,” Ph.D. Thesis, Univer that may increase the risk for infection and autoimmune sity of Utah, Salt Lake City, Utah, 1990). Also, 6-MBOA reactions following a variety of Stressful Stimuli, including may increase the rate of Synthesis of follicle Stimulating injury, Surgery, infection, psychological StreSS, mental ill hormone (Butterstein et al., “The plant metabolite 6-meth oxybenzoxazolinone interacts with follicle-Stimulating hor ness (e.g., depression), and physical challenges Such as mone to enhance ovarian growth, Biology of Reproduction, intense elite athlete training. The resulting complications 39:465-71, 1988) and pituitary prolactin (Vaughan et al., due to immune impairment in humans may facilitate Sig "Hormonal consequences of Subcutaneous 6-methoxy-2- nificant increased costs of medical care and lost work dayS. benzoxazolinone pellets or injections in prepubertal male 0019. Those skilled in the art, including ethnobotanists, and female rats, Journal of Reproduction and Fertility, pharmacognosists, medicinal chemists, and certain mental 83:859-66, 1988). In addition, hypothalamic luteinizing health care practitioners are constantly in Search of new hormone-releasing hormone contents and pituitary gland compounds derived or isolated from plant materials that weights are greater for at least one rodent Species implanted have anti-anxiety properties. For example, anti-anxiety with capsules containing 6-MBOA (Urbanski et al., “Influ activity has been widely associated with kava kava and ence of photoperiod and 6-methoxybenzoxazolinone on the commercial products containing kava kava and claiming and reproductive axis of inbred LSH/SS Lak male ham anti-anxiety effects are widely known. sters.''Journal of Reproduction and Fertility, 90:157-163, 0020. An object of the present invention is to develop 1990). compositions and methods for calming and reducing anxiety US 2005/0250772 A1 Nov. 10, 2005 in animals and humans entailing use of compounds belong tions to reduce levels of catecholamine compounds (e.g., ing to related chemical families, of which 6-MBOA is a epinephrine and norepinephrine) that may be increased due member. Pursuant to this end, a further object is to develop to anxiety. methods for extracting, deriving, and/or isolating Said com 0030. It is a still further object of the present invention to pounds from plant and animal Sources in amounts Suitable provide novel chemical compositions derived, isolated, and/ for human therapeutic use. or extracted from monocotyledonous plants or by chemical 0021 While past research by those skilled in the art has Synthesis, and methods for using Said compositions to affect attempted to isolate, identify, and characterize new plant gamma-aminobutyric acid (GABA) levels in a fashion asso compounds with anti-anxiety properties, 6-MBOA has here ciated with the reduction or elimination of anxiety. tofore previously not been identified or evaluated for calm ing and anti-anxiety properties. Therefore, novel compounds 0031. It is a still further object of the present invention to derived, isolated, extracted, produced, and/or harvested provide novel chemical compositions derived, isolated, and/ from monocotyledonous plants or by chemical Synthesis and or extracted from monocotyledonous plants or by chemical methods for using the same to reduce anxiety and related Synthesis, and methods for using Said compositions to affect pathology in animals and humans would be a significant the function of gamma-aminobutyric acid (GABA) recep advancement in the art. Such novel compositions and meth tors (e.g., allosteric modification of the receptor to increase ods are disclosed and taught herein. duration and/or frequency of ion channel opening). 0032. It is a still further object of the present invention to BRIEF SUMMARY AND OBJECTS OF THE provide novel chemical compositions derived, isolated, and/ INVENTION or extracted from monocotyledonous plants or by chemical 0022. A primary object of the present invention is to Synthesis, and methods for using Said compositions to provide novel chemical compositions derived, isolated, and/ decrease comorbid Symptoms related to anxiety. or extracted from monocotyledonous plants or by chemical 0033. It is a still further object of the present invention to Synthesis, and methods for using Said compositions to aid in provide novel methods for growing and harvesting mono calming humans and animals experiencing anxiety during cotyledonous plants to obtain phenolic compounds with a periods of StreSS or due to anxiety conditions. phenolic molecule to which are covalently linked an oxy gen-containing group, a nitrogen or Second OXygen contain 0023. It is also an object of the present invention to ing group, and at least one C-C alkoxy group. provide novel chemical compositions derived, isolated, and/ or extracted from monocotyledonous plants or by chemical 0034. Additionally, it is an object of the present invention Synthesis, and methods for using Said compositions to to provide novel methods for preserving and augmenting elevate levels of indolamines, for example, Serotonin and innate immune defenses in humans and animals by admin melatonin in humans and animals. istering phenolic compounds with a phenolic molecule to which are covalently linked an oxygen-containing group, a 0024. It is a further object of the present invention to nitrogen or Second oxygen containing group, and at least one provide novel chemical compositions derived, isolated, and/ or extracted from monocotyledonous plants or by chemical C-C alkoxy group. Synthesis, which function as Serotonin and/or melatonin 0035) It is another object of the present invention to analogs and/or agonists in mammals. provide novel methods for calming and/or reducing anxiety in humans and animals by administering phenolic com 0.025. It is a still further object of the present invention to pounds with a phenolic molecule to which are covalently provide novel methods for growing monocotyledonous linked an oxygen-containing group, a nitrogen or Second plants which results in an increased yield of 6-MBOA or OXygen containing group, and at least one C-C alkoxy other phenolic and indoleamine compounds. grOup. 0026. In addition, it is an object of the present invention 0036. It is also an object of the present invention to to provide novel methods for harvesting monocotyledonous provide novel methods for growing and harvesting mono plants which are efficient for producing 6-MBOA or other cotyledonous plants to obtain precursors of phenolic com phenolic and indoleamine compounds. pounds comprising benzoxazinoids-cyclic hydroxamic acid, 0.027 Moreover, it is an object of the present invention to lactams, and their corresponding glucosides. provide novel chemical compositions derived, isolated, and/ 0037 Additionally, it is an object of the present invention or extracted from monocotyledonous plants or by chemical to provide novel methods for preserving and augmenting Synthesis, and methods for using Said compositions for innate immune defenses in humans and animals by admin Suppressing the anxiety eliciting effects of the hypothalamic istering precursors of phenolic compounds comprising ben pituitary-adrenal (HPA) axis. ZOxazinoids-cyclic hydroxamic acid, lactams, and their cor 0028. It is also an object of the present invention to responding glucosides. provide novel chemical compositions derived, isolated, and/ 0038 Consistent with the foregoing objects, and in accor or extracted from monocotyledonous plants or by chemical dance with the invention as embodied and broadly described Synthesis, and methods for using Said compositions for herein, it has been found that certain phenolic compounds counteracting detrimental behaviors associated with anxiety. and precursors of phenolic compounds, related to each other 0029. Additionally, it is an object of the present invention by Shared Structural Similarities and having Structural Simi to provide novel chemical compositions derived, isolated, larities with Serotonin and/or melatonin, are effective for and/or extracted from monocotyledonous plants or by reducing Symptoms of anxiety in humans and animals. AS chemical Synthesis, and methods for using Said composi appreciated, the novel compounds of the present invention US 2005/0250772 A1 Nov. 10, 2005 may naturally exist as plant Secondary metabolites in the 0043 FIG. 1 is a bar chart illustrating a cellular immune earlier growth of monocotyledonous plants, become con response to dinitrofluoro-benzene (DNFB) resulting from a centrated from their ingestion within certain animal parts, or Study examining ear Swelling in Stressed and non-stressed may be Synthesized by chemical means. The present inven mice exposed to one presently preferred embodiment of tion contemplates the therapeutic use of novel compounds compounds of the present invention and the corresponding derived, isolated, and/or extracted from monocotyledonous plants or by chemical Synthesis for use in the reduction of Stressed and non-stressed control mice; anxiety in humans and animals. 0044 FIG. 2 is a bar chart illustrating an evaluation of 0039. One presently preferred embodiment of a therapeu the effects of one presently preferred embodiment of com tic method of the present invention comprises the ingestion pounds of the present invention on preservation of cell by human or animal of the novel compounds of the present mediated immunity (CMI), manifested in response to dini invention over a certain period of time, or other means for trofluorobenzene (DNFB) resulting from a study examining getting the compounds of the present invention into the ear Swelling in Stressed and non-stressed mice exposed to body. Both males and females benefit from ingesting the one presently preferred embodiment of the compounds of compounds of the invention, while Still contained in dried leaves from monocotyledonous plants with Such compounds the present invention (i.e., at concentrations of 40 mg/kg, 25 or taken as purified and/or Synthesized preparations. It mg/kg, and 5 mg/kg) and the corresponding stressed and appears, based on the findings of the inventors that the novel non-stressed control mice; compounds of the present invention act as indoleamine 004.5 FIG. 3 is a bar chart illustrating one presently analogs and/or agonists and assist in reducing anxiety preferred embodiment of a method of the present invention related to StreSS. showing a he cellular immune response to dinitrofluoroben 0040 AS discussed in greater detail hereinbelow, one Zene (DNFB) resulting from a study examining ear Swelling presently preferred embodiment of the present invention in stressed calves exposed to two (2) different dosages of one comprises the administration of phenolic compounds presently preferred embodiment of the compounds of the belonging to related chemical families of which 6-methoxy present invention and corresponding Stressed and non 2,3-benzoxazolinone (6-MBOA) is a member. These phe Stressed control calves, and nolic compounds Share a structural Similarity with melatonin and indoleamine compounds. Based on Structural Similari 0046 FIG. 4 is a bar chart illustrating one presently ties with melatonin, compounds of the present invention preferred embodiment of a method of the present invention were Studied for their effects on Symptoms related to anxiety. showing the cellular immune response to dinitrofluoroben In one presently preferred embodiment of a method of the Zene (DNFB) resulting from a study examining neck swell present invention, particular emphasis is placed on calming ing in stressed calves exposed to two (2) different dosages of and reducing anxiety and related behaviors in humans and one presently preferred embodiment of the compounds of animals by the administration of a therapeutically effective the present invention and corresponding Stressed and non amount of a composition of 6-MBOA or pharmaceutically Stressed control calves. acceptable Salts thereof. 0041) A source of the compounds of the present invention DETAILED DESCRIPTIONS OF THE for use in reducing anxiety in humans and animals may be PREFERRED EMBODIMENTS found in certain monocotyledonous plants in their early growth stages. To obtain the compounds of the present 0047. It will be readily understood that the components of invention at concentrations Suitable for human therapeutic the present invention, as generally described and illustrated use from these Such monocotyledonous plants, the harvest in the Figures, formulae and tables herein, could be arranged ing of these plants at an early life history Stage and drying and designed in a wide variety of different configurations. using explicit parameters, as well as Specific analytical Those of ordinary skill in the art will, of course, appreciate criteria to ascertain Suitability, are employed. However, it is that various modifications to the details herein may be made also possible to get the compounds of the present invention without departing from the essential characteristics of the at concentrations Suitable for human therapeutic use from invention, as described. Thus, the following more detailed animal parts, including, but not necessarily limited to, the description of the embodiments of the compositions and velvet antler tips of deer and elk (Cervidae), where they methods of the present invention, as represented in FIGS. 1 become concentrated after ingestion by the animal of Sprout ing and otherwise immature grasses. Furthermore, the com through 4, is not intended to limit the scope of the inven pounds of the present invention may also be obtained tion, as claimed, but it is merely representative of the through chemical Synthesis. presently preferred embodiments of the invention. BRIEF DESCRIPTION OF THE DRAWINGS 0048 Referring to Formulae I-IV below, the compounds 0042. The foregoing and other objects and features of the of the invention have in common a phenol molecule to present invention will become more fully apparent from the which are covalently linked an oxygen-containing group, a following description and appended claims, taken in con nitrogen- or another oxygen-containing group, and a C-C, junction with the accompanying drawings. Understanding alkoxy group. Using Standard conventions for depicting that these drawings depict only typical embodiments of the chemical Structures, Formulas I-III disclose the chemical invention and are, therefore, not to be considered limiting of Structures and Specific parameters defining the compounds its scope, the invention will be described with additional of the invention. Formula IV is a unifying formula that Specificity and detail through use of the accompanying depicts all the presently preferred embodiments of the drawings in which: compositions of the present invention. US 2005/0250772 A1 Nov. 10, 2005

0049. Formula I-A chemical composition according to 0063. Wherein, “R” representsp C-C1 - 4 alkoxy,y with the the formula proviso that R is in the 4 or 5 ring position; 0064. Wherein “n” represents one of the integers 0, 1 or 2, 0065. Wherein “B” represents H and “A” represents -OH, -NH, or NHCR, where R' denotes C-C, alkyl; and “BA” represents

0050. Wherein “R” represents C-C alkoxy, with the O H OH O OH proviso that R is in the 4 or 5 ring position; | | || | 0051) Wherein “n” represents one of the integers 0, 1 -C-N- or ---- or 2, H 0052) Wherein “A” represents -OH, -NH, or NHCR', where R' represents C-C alkyl; 0066 or pharmaceutically acceptable salts thereof. 0053 or pharmaceutically acceptable salts thereof. 0067. It has been found that compounds of the invention, when ingested or otherwise introduced into the body of a 0.054 Formula II-A chemical composition according to human or animal, are effective as: (1) an antidepressant in the formula that it betters mood and reduces or relieves Symptoms of depression; (2) in Sex therapies in the Sense that it improves Sexual desire and performance (e.g., desire, arousal, and performance); (3) an effective adjunctive therapy for weight loss; (4) as an adjunctive treatment for Substance abuse and addiction; (5) for preserving and augmenting innate immune defenses in humans and animals; and (6) for reducing Symptoms and comorbid behaviors relating to anxiety. The compounds may be administered in the form of ground parts of plants in which they naturally occur Such as, by way of 0055 WhereinCC “R” representsrep C-C alkOXy,alkoxy, Withwith theth example but not by limitation, the ground leaves of imma proviso that R is in the 5 or 6 ring position; ture corn, or as purified or chemically Synthesized com 0056. Wherein “n” represents one of the integers 0, 1 pounds in a pharmaceutically acceptable carrier. or 2, 0068 The compounds of the present invention may be 0057 or pharmaceutically acceptable salts thereof. administered orally in the form of tablets, capsules, Suspen Sions, Solutions, or other means Suitable for Such ingestion, 0.058 Formula III-A chemical composition according to perhaps as an admixture with other compounds to enhance the formula absorption into the blood Stream or to otherwise assist in achieving the desired effects. Likewise, oral administration is contemplated to include Sublingual (i.e., under the tongue) dosage forms. The compounds of the present invention may also be delivered by intranasal (i.e., through the nasal Structures) or transmucosal (i.e., across mucous membranes) administration. 0069. In addition, the compounds of the present invention may also be administered parenterally, as a Subcutaneous, intramuscular, or intravenous injection, or by way of an 0059 Wherein “R” representsp C-C1 - 4 alkoxy,y with the implant for Sustained release. When administered parenter proviso that R is in the 6 or 7 ring position; ally, the compounds of the present invention are to be dissolved in physiologically acceptable liquid media and/or 0060. Wherein “n” represents one of the integers 0, 1 otherwise compounded in accordance with the known phar or 2, maceutical art. Another mode of administering the com 0061 or pharmaceutically acceptable salts thereof. pounds of the present invention may be a transdermal patch, in which entry of the compounds of the invention into the 0.062 Formula IV-A chemical composition according body is facilitated via acceptable and appropriate carrier to the formula molecules. 0070. Unless otherwise defined, the technical, scientific, and medical terminology used herein has the Same meaning as understood by those informed of the art to which this invention belongs. However, for the purposes of establishing Support for various terms that are used in the present application, the following technical comments, definitions, and review are provided for reference. US 2005/0250772 A1 Nov. 10, 2005

0071. “Stress” may be defined as the failure of an animal 0083) “Macrophage” may be defined as a large phago or human to adapt to a stimulus or factor (e.g., physical, cyte. Wherein a phagocyte, typically a leucocyte or reticu chemical, or emotional (as trauma, histamine, or fear, loendothelial cell, may be any cell that characteristically respectively)) and which causes physiologic tension that engulfs foreign material. Phagocytes may function in the may predispose the animal or human to development of a body to remove and consume debris and foreign bodies (e.g., disease or otherwise impair normal physiological function. degenerating tissue or bacteria). 0.072 “Cytokines' may be defined as a class of immu 0084) “Complement” may refer to any thermolabile (i.e., noregulatory proteins (e.g., as interleukin, tumor necrosis inactivated by excessive heat) Substances normally found in factor, or interferon) that are Secreted by cells. Cytokines are blood Serum and plasma. Complement may work in com primarily Secreted by cells associated with the immune bination with antibodies to cause the destruction of bacteria, System. foreign blood corpuscles, and other antigens. 0.073 “Hormone' may be defined as a chemical product 0085) “Antibody” may refer to any of various body (often an organic chemical) of living cells that is Secreted globulin proteins which may be produced by cells of the into body fluids where it may be transported to other areas Specific immune System in response to infection or admin of the body and produce a specific effect on the activity of istration of Suitable antigens or haptens (i.e., non-antigenic cells. or weakly antigenic Substance) and combine with the anti 0.074 “Immune system” may be defined as the system gens (e.g., bacteria, toxins, or foreign red blood cells) to responsible for protecting the body of an animal or human neutralize them and/or neutralize their toxins. from foreign Substances, cells, and tissues. The immune System may include the thymus, Spleen, lymph nodes, Spe 0086) Many animals and all humans have an immune cial deposits of lymphoid tissue (e.g., as in the gastrointes System for providing cellular and/or physiological defense tinal tract and bone marrow), antibodies, and lymphocytes mechanisms against foreign particles, for example and not (e.g., B cells and T cells). by way of limitation, invading microbes and microorgan isms (e.g., bacteria, Viruses, amoeba, parasites, and the like), 0075 “T-cell” may be defined as any of several lympho foreign proteins and carbohydrates, and other antigenic (i.e., cytes that may undergo differentiation in the thymus. T-cells Stimulation of antibody and/or complement response) Sub may possess highly specific cell-Surface antigen receptors, stances. The term “immunity' is often used interchangeably and may be involved in controlling the initiation or Sup pression of cell-mediated and humoral immunity. T-cells with immune system and its role in the body. may also possess receptors that lyse antigen-bearing cells. 0087. The term “inflammation” may generally refer to the 0076) “Cell mediated immunity (CMI)” may be defined process of the immune System in identifying a foreign as relating to the immune response mediated primarily by particle, antigen, or damaged tissue, mobilizing cells, and molecules to attack the foreign particle, antigen or damaged T-cells and especially cytotoxic T-cells rather than by anti tissue and disabling and/or destroying the foreign particle or bodies secreted by B-cells. antigen or eliminating damaged tissue. Inflammation often 0.077 “Catecholamines' may be defined as any of various may be characterized by capillary dilatation, infiltration with chemical compounds (e.g., epinephrine, norepinephrine, leukocytes (i.e., white blood cells), localized and/or gener dopamine, and the like) that may contain a benzene ring with alized temperature increase, and localized and/or general two adjacent hydroxyl groups and a side chain of ethy ized pain. Capillary dilatation and infiltration with leuko lamine. Catecholamine may function as hormones or neu cytes may be further characterized as edema (i.e., Swelling) rotransmitters or, alternatively, may function as both. around the Site of infection and/or damaged tissue. Inflam 0078 “Corticoid” may be defined as any of various mation may be an important mechanism for animals and organic chemical compounds which may be extracted from humans to disable, destroy, and/or eliminate toxic particles the adrenal cortex. Corticoid may commonly refer to Ste or damaged tissue. roids or corticosteroids, several of which are hormones (e.g., 0088 Those skilled in the art may appreciate that the corticosterone, cortisone, and aldosterone). immune System is a highly complex organization of cells 0079) “Glucocorticoid” may refer to a corticoid that has and organs which are spread throughout the body. Moreover, a primary effect on carbohydrate metabolism (e.g., corti two general divisions of immunity are commonly recog Sone, hydrocortisone, prednisone, prednisolone, triamcino nized by those skilled in the art. Natural (i.e., innate or lone, methylprednisolone, dexamethasone, betamethasone, native) immunity may be largely dependent on the comple and the like). ment System, phagocytes and cytokines to protect the body 0080) “Innate” may be defined as existing in or belonging from infection. Animals and humans may be born with to a perSon, animal, or other living organism from birth. natural immunity which is encoded into genes. Innate may be used Synonymously with native or natural. 0089 Complement may be a group of related proteins 0081) “Immunity” may refer to the ability to resist a that assist natural immunity cells and antibodies in recog particular disease, especially by preventing the development nizing (i.e., a cognitive phase) antigens or foreign particles. of a pathogenic microorganism or by counteracting the Complement proteins may bind to the Surface of an antigen effects of pathogenic microorganism toxins and cellular or foreign particle (i.e., activation phase). Activation by products. complement proteins may increase the membrane porosity 0082 “Natural immunity” may refer to immunity pos of the antigen or foreign particle, which in turn may lead to Sessed by a group (i.e., a race, Strain or Species) and disabling the antigen or foreign particle or, in the alternative, occurring in an individual as part of its genetic and/or may serve as a "tagging mechanism to enhance identifica biological makeup. tion of the antigen or foreign particle by other natural US 2005/0250772 A1 Nov. 10, 2005 immunity cells (e.g., phagocytes) and/or antibodies. Natural glands. The pineal gland may be responsible for the Synthe immunity cells and/or antibodies may then disable and/or sis of melatonin from Serotonin. Melatonin may have mul eliminate the antigen or foreign particle (i.e., effector phase). tiple effects on the body, which may include Stimulating 0090 Phagocytes may belong to a group of cells which and/or enhancing the function of leukocytes and/or lympho engulf and destroy other cells or particles through a proceSS cytes in response to an antigen or foreign particle. Serotonin commonly referred to as phagocytosis. In addition, phago may also have a positive effect on the immune System by cytes (e.g., macrophages, neutrophils, natural killer cells) Stimulating the proliferation of T-cells and may enhance the may Secrete Soluble mediators, commonly referred to as function of certain phagocytes (e.g., macrophages). cytokines, which may be active in disabling and/or destroy 0097. The hypothalamus, pituitary, and adrenal glands ing an antigen or foreign particle. Cytokines may include for form an important hormonal feedback pathway the, hypo example, and not by way of limitation, interferons (e.g., C.- thalamic-pituitary-adrenal (HPA) axis, for regulating the and B-interferons), tumor necrosis factors (TNF), and inter response of the body to StreSS. The hypothalamus may leukins. Secrete hormones which may act upon the pituitary gland 0.091 Specific (i.e., acquired) immunity may retain the causing it to release its hormones. Hormones from the use of cells and molecules of natural immunity and may add pituitary gland may have actions on endocrine or other two additional immune defense features. Specific immunity organs in the body. The adrenal glands may be responsible may provide a host (i.e., animal or human) with the ability for producing a plurality of Steroid hormones, including to “remember a specific antigen or foreign particle. This glucocorticoids, mineralocorticoids, and androgens. The memory property may allow the host to mount a more adrenal glands may also be responsible for producing cat effective defense upon each Subsequent encounter with the echolamine hormones (e.g., epinephrine, norepinephrine, Specific antigen or foreign particle. Vaccination may be dopamine, and the like). based upon the property of Specific immunity to remember 0098. The higher incidence of infectious diseases in a specific antigen or foreign particle. humans and animals Subjected to Stressful situations may be 0092 Specific immunity may also provide a host with the a result of the neuroendocrine and immune changes that ability to “amplify a natural immunity defensive response occur during a challenging period. These changes may be to an antigen or foreign particle. Moreover, the amplification the result of adaptation to the Stressful situation. A widely proceSS may also involve more accurate targeting of natural investigated physiological and anatomical component of this immunity to the site of entry for an antigen or foreign process of adaptation to StreSS is the HPA axis. Activation of particle. the HPA axis usually results in the elevation of glucocorti coid levels associated with StreSS, illness, and injury. Glu 0093. In addition, specific immunity may utilize a cog cocorticoids may have Suppressive effects on immune func nitive phase where specific antigens or foreign particles may tion and may play a role in StreSS-associated immune be identified; an activation phase where specific antigens or Suppression. Immunosuppressive effects of glucocorticoids foreign particles may be tagged; and an effector phase may therefore include reduction in the production of and/or wherein Specific antigens or foreign particles may be acted destruction of immune System cells and Suppression of upon by an antibody, lymphocyte, or lymphocyte-derived inflammation by the immune System. In addition, adrenal cytokine to effectively disable and/or destroy the antigen or catecholamines (e.g., epinephrine and norepinephrine) may foreign particle. be elevated in response to StreSS, and may have a prominent 0094 Natural and specific immunity may be different in role in StreSS-related emotional States, including anxiety. that complement and phagocytes are unable to distinguish 0099 Albeit with structural similarities to melatonin, different antigens and may not be enhanced in numbers 6-MBOA and related compounds may function as melatonin following exposure of the host to the antigen or foreign agonists and at the C- and B-adrenergic cell receptors in their particle. Whereas antibodies and lymphocytes may be able own right. Whereas melatonin exacerbates Symptoms of to identify specific antigens or foreign particles and may dysphoria in depressed people, 6-MBOA and related com undergo an amplification in number following exposure of pounds of the present invention, as a melatonin agonist, the host to the antigen or foreign particle. Works in contrary fashion and actually Stimulates a better 0.095 Two classifications of specific immune responses mood. may be recognized depending on which components of the 0100 AS melatonin agonists, novel compounds of the immune System are being utilized. Humoral immunity may present invention may stimulate the production of melatonin refer to the action of antibodies and other molecules which or may enhance the actions of melatonin on the immune eliminate and/or neutralize antigens. Cell-mediated immu system. For example, and not by way of limitation, 6-MBOA nity (CMI), Sometimes referred to as cellular immunity, may and related compounds of the present invention may stimu refer to the action of lymphocytes (e.g., B-lymphocytes late the production of lymphocytes and the production of (“B-cells”) and T-lymphocytes (“T-cells”)), in eliminating cytokines by immune System cells. and/or neutralizing antigens or foreign particles. 0101 There has previously been no suggestion to impli 0.096 Those skilled in the art will appreciate that endo cate the administration of 6-MBOA and related novel com crine and hormonal regulation of the immune System may pounds of the present invention in preserving and/or aug involve a complex process. A plurality of hormones and menting immune System function and/or reducing anxiety hormonal pathways may be involved in immune System and related comorbidities, except for depression. To this end, regulation. Important endocrine glands in immune regula the inventors of the present invention have found that tion may include for example, and not by way of limitation, calming and reducing Symptoms of anxiety may be accom the pineal gland, hypothalamus, pituitary gland, and adrenal plished through use of the novel compounds of the present US 2005/0250772 A1 Nov. 10, 2005

invention to Stimulate Serotonin- and melatonin-induced contemplated, however, that the Spirit and Scope of the effects on the immune System and/or reduction of StreSS present invention includes all anxiety conditions and comor induced neuroendocrine products. bid conditions. 0102) In addition, serotonin and melatonin both may have immunoregulatory properties. Serotonin may enhance cell 0115) Anxiety disorders may be the most common type of human psychiatric disorder in the United States. Anxiety proliferation and macrophage antigen presentation. Melato disorders may include, for example and not by way of nin may have enhancing activities in a number of StreSS limitation, panic disorder, obsessive-compulsive disorder, induced immune dysfunction models, including, but not post-traumatic StreSS disorder, Specific phobia, Social pho limited to, restraint StreSS, injury, and cortisol administra bia, and generalized anxiety disorder. Anxiety disorders may tion. Moreover, as appreciated by those skilled in the art, be characterized by uneasiness, fearfulness, and distress, 6-MBOA or related compounds of the present invention may Sometimes experienced without clear evidence of real be structurally and/or functionally similar to melatonin and threats. During situations that are stressful, humans and catecholamines (e.g., epinephrine, alpha- and beta-adrener animals may respond with alterations in behavior and auto gic compounds, and the like), respectively. nomic and hormonal functions and Secretions. These responses may result in exacerbation of anxiety Symptoms 0103 6-MBOA or related compounds of the present during Situations of StreSS, affecting mental and physical invention may have both inhibitory and stimulatory effects health and performance. Increased arousal during periods of in and around the central nervous System. As a melatonin anxiety may result in hypervigilance, increased Startle analog, 6-MBOA may work at melatonin receptors as a response, impaired concentration and learning, insomnia, melatonin antagonist, thus blocking the negative effects of irritability, all of which may lead to decreased performance in a variety of Settings in humans and animals. Millions of melatonin on Sexual maturation and/or function. In addition, people in the United States (i.e., greater than 23 million the beta-adrenergic receptor effect of 6-MBOA may help to diagnosed) may Suffer from these conditions with significant deSensitize melatonin receptors and thus minimize the nega costs to individual performance and health, lost productivity, tive effects of melatonin on Sexual maturation and/or func and costs of treatment. tion. Therefore, work by the inventors demonstrate that presently preferred embodiments of the compounds of the 0116 Performance and health of animals may often be present invention possess Some of the positive effects of challenged and detrimentally affected by anxiety behaviors. Anxiety provoking situations of novel environments, ship melatonin function (e.g., calming and/or anti-anxiety effects ping and transport, illness or injury, Weaning, crowding, and preserving and/or augmenting immune function) with castration, Vaccination, dipping, deworming, tagging, brand out Stimulating the negative effects of melatonin action (e.g., ing, shoeing, and other processing procedures including inhibiting sexual maturation, dysphoria, and the like). training and performance may have detrimental effects on 0104. One presently preferred embodiment of a novel health, growth, and performance. Extensive effort in training individual animals to reduce anxiety is costly and often may method of the present invention for the prevention or not be practical. Similarly, loSS of weight due to anxiety treatment of anxiety disorders and related behaviors and related StreSS conditions may result in heavy financial loSS to States involves administering to an animal or human in need the animal industry. of Such treatment an effective amount of a compounds described in present application. There are Several distinct 0117 Currently available methods for treating anxiety anxiety disorders and related behaviors and States, including disorders may include, for example and not by way of for example and not by way of limitation: limitation, psychologic therapy, including behavioral, cog nitive and emotive, and relaxation techniques, or training in 01.05 Panic Disorder animals. These methods may involve a significant amount of time and expense to achieve reduction in anxiety and 0106 Agoraphobia enhancement of performance. Medications may also be used in humans and animals to reduce anxiety, including, for 0107 Generalized Anxiety Disorder (GAD) example and not by way of limitation, Serotonin reuptake 0108) Specific Phobia inhibitors (i.e., fluoxetine, paroxetine, Sertraline, citalopram, escitalopram, fluvoxamine, Venlafaxine, etc.), benzodiaz 0109) Social Phobia epines (e.g., alprazolam, chlordiazepoxide, cloraZepate, diazepam, halazepam, lorazepam, oxazepam, etc.), beta 0110. Obsessive-Compulsive Disorder (OCD) blockers (i.e., beta-adrenergic receptor antagonists (e.g., propranolol, etc.), monoamine oxidase inhibitors (e.g., phe 0111 Post-Traumatic Stress Disorder (PTSD) nylzine, tranylcypromine, etc.), azapirones (e.g., buSpirone), 0112 Acute stress disorder diphenylmethanes (e.g., diphenhydramine, hydroxyzine, etc.), and tricyclic antidepressants (e.g., amitryptyline, clo 0113 Attention-Deficit Hyperactivity Disorder mipramine, doxepin, imipramine, trimipramine, (ADHD) desipramine, nortriptyline, protriptyline, etc.). These medi cations may have Serious Side effects may not be easily 0114. The diseases described above as well as other accessible for the treatment of animals, and may be inap anxiety disorders are classified in the Diagnostic and Sta propriate for food production Species. Therefore new meth tistical Manual of Mental Disorders, 4th Version, published ods for treating anxiety disorders are needed for both by the American Psychiatric Association (DSM-IV). It is humans and animals. US 2005/0250772 A1 Nov. 10, 2005

0118. The present invention provides novel compositions present invention, as represented in Examples 1-17, is not and methods for the prevention or treatment of anxiety intended to limit the Scope of the invention, as claimed, but disorders which involves administering to an animal or is merely representative of presently preferred embodiments human in need of Such treatment an effective amount of a of the invention. compound described in this application. In addition, behav iors and negative consequences in animals related to anxiety may include, for example and not by way of limitation: (1) EXAMPLE 1. repetitive, compulsive and Stereotypic actions (e.g., licking skin to ulceration, cribbing, Sucking air, pacing, and other The Similar Physiological Effects of the pathologic behaviors); (2) fear and phobias related to han Compounds of the Invention dling (e.g., loading into trailers, being touched, training, performance); (3) weaning stress; (4) Social StreSS/mixing 0.122 Representative compounds of Formulas I, II, and novel animals; (5) transportation; (6) confinement and III are shown below as compounds 1-7. The compounds of crowding with asSociated weight loSS and negative health Formulas I, II and III have like physiological properties, and effects; and (7) in food production animals (i.e., poor meat as Such may be considered as Similar or equivalent for quality due to stress responses). Any of these disorders may therapeutic purposes, and were tested via a rodent model. be treated or prevented by the method and compounds of the present invention. Further, Stressful Situations that may place 0123 1. 2-amino-5-methoxyphenol Member of For individual humans into novel, challenging, or threatening mula I positions may also elicit anxiety Symptoms and related behaviors and the present invention may minimize detri mental effects of Such anxiety producing Situations and enhance recovery, and may also prevent or minimize estab lishment of longer term anxiety disorders. Effective treat CHO, r ment of anxious humans and animals may aid in learning, training, and performance and may minimize or prevent NH2 detrimental physical consequences of StreSS and anxiety. 0119 While past research by those skilled in the art has attempted to isolate, identify, and characterize new plant compounds with immune System, anti-anxiety and mood 0.124 2. 6-methoxy-2-benzoxazolinone Member of Stimulating properties, 6-MBOA and are related compounds Formula II of the present invention have heretofore previously not been identified or evaluated for preserving and/or augmenting innate immune defenses and/or Stimulating mood. There CHO O fore, as readily appreciated by those skilled in the art, novel N compounds derived, isolated, extracted, produced, and/or CEO harvested from monocotyledonous plants and methods for M using the same to preserve and/or augment innate immune defenses in humans and animals would be a significant H advancement in the art. Such novel compositions and meth ods are disclosed and taught herein. 0125 3. 2,4-dihydroxy-7-methoxy-1,4-(2H)-benzox 0120 While past research by those skilled in the art has azin-3-one Member of Formula III attempted to isolate, identify, and characterize new plant compounds with calming and/or anti-anxiety properties, 6-MBOA has notheretofore been identified or evaluated for CHO On OH calming or reducing anxiety. The novel compounds derived, isolated, extracted, produced and/or harvested from mono cotyledonous plants or, in the alternative, by chemical O y1SC Synthesis and methods for using Said compounds to calm rs. and/or reduce anxiety and related comorbid behaviors and OH States in animals and humans as contemplated by the present invention is therefore a significant advancement in the art. 0121 The following examples will illustrate the practice 0.126 4.2-hydroxy-4-methoxyacetanilide Member of of the present invention in further detail. It will be readily Formula I understood by those skilled in the art that the following methods, formulations, and compositions of novel com pounds of the present invention, as generally described and CHO OH illustrated in the Example herein, are to be viewed as exemplary of the principles of the present invention, and not as restrictive to a particular structure or process for imple NH- i-Cit menting those principles. Thus, the following more detailed O description of the presently preferred embodiments of the methods, techniques, formulations and compositions of the US 2005/0250772 A1 Nov. 10, 2005

0127 5. 2-hydroxy-4-ethoxyacetanilide Member of Formula I TABLE 1-continued Effect of injecting compounds of the invention, members of Formulas I, II and III on uterine weight in the montane vole. Microtus montanus. Number Average Formula of Uterine NH-C-CH Compound Injected Numeral Animals Weight (mg) 2-hydroxy-4-ethoxyacetanilide I 8 22.2 3.98 2-amino-5-methoxyphenol I 8 21.8 3.5* 2-hydroxy-5-methoxyacetanilide I 8 21.1 - 4.4*** 0128 6. 5-methoxy-2-benzoxazolinone Member of 2-amino-4-methoxyphenol I 8 22.2 3.2* Formula III *Significantly different from control at P < 0.001 ** Significantly different from control at P < 0.015 *** Significantly different from control at P = 0.004 0.132. On average, the uterine weight in voles receiving these was 22.8 g., fifty percent (50%) greater than for the CHO control group. The greatest average weight increase, eighty two percent (82%) more than the uterine weight for the control Voles, was in those females administered 6-methoxy 2-benzoxazolinone, but even the least effect of a compound 0129. 7. 2-hydroxy-5-methoxyacetanilide Member of belonging to Formulas I, II or III, that for 5-methoxy-2- Formula I benzoxazolinone, entailed a thirty-two percent (32%) increase in uterine weight. The results show that physiologi cal effects or modes of action are held in common by the OH compounds of the invention.

EXAMPLE 2 CHO NH-C-CH I Compounds of the Invention as an Antidepressant and Aphrodisiac in Human Males 0130. Female montane voles, Microtis montanus, 0133. This component of the invention relates to a received intra peritoneal injections of representative com method for lessening depression and otherwise bettering pounds belonging to Formulas I, II and III for three con mood or feelings of well-being, Said method comprising the secutive days and Sacrificed twenty-four (24) hours after the administration to human males of an effective amount of one last injection to examine uterine weight response. To assess or more of the compounds of the invention defined herein. the properties of each representative compound, pure ones This component of the invention also relates to a method for made by chemical means (University of Utah Department of treating Sexual dysfunction or otherwise increasing Sexual Chemistry, Salt Lake City, Utah) were prepared specifically desire and performance, including but not necessarily lim for this test. All compounds were injected at a dose level of ited to lacking interest in SeX, problems with arousal, not 5 mcg/day, dissolved in five percent (5%) propylene glycol enjoying Sex, and anxiety about Sexual performance, Said for a total injection volume of 0.5 ml. Control animals method comprising the administration of an effective received 0.5 ml of five percent (5%) propylene glycol only. amount of one or more of the active compounds of the All voles were 4-5 weeks old and weighed 25-29 g. invention. 0131. As illustrated in Table 1 below, all compounds 0.134. A double-blind crossover study was done on belonging to Formulas I, II and III caused a Statistically human males to test compounds of the invention as a Significant increase in uterine weights. therapeutic agent for treating depression or otherwise elevat ing mood as well as bettering Sexual function. The trial had TABLE 1. three phases, each two weeks in duration, during which Effect of injecting compounds of the invention, participants took compounds of the invention for one phase members of Formulas I, II and III on uterine weight or two weeks. The daily dose was made up from compounds in the montane vole. Microtus montanus. of the invention naturally contained in the ground leaves Number Average from immature corn plants, 30-45 cm tall, standardized with Formula of Uterine Synthesized 6-methoxy-2-benzoxazolinone, to a total of 15 Compound Injected Numeral Animals Weight (mg) mg 6-methoxy-2-benzoxazolinone. A dose of fifteen milli Control (Propylene Glycol) 8 15.2 + 2.4 grams (15 mg) was selected because this was considered a 6-methoxy-2-benzoxazolinone I 11. 27.7 5.6* likely minimum effective daily amount for humans, extrapo 5-methoxy-2-benzoxazolinone III 8 20.0 - 4.7** lated from prior Studies on rodents, rabbits and other ani 2-hydroxy-4-methoxyacetanilide I 8 23.1 2.7* mals. Previous anecdotal trials on humans done by the inventors Suggested that a 15 mg daily dose had a desirable US 2005/0250772 A1 Nov. 10, 2005 effect, but no adverse consequences to health. In general, therapeutically effective amounts of compounds of the TABLE 2-continued invention may be found in a daily dosage range of between HAD and ASEX summaries for administration of about 5 micrograms (mcg) to about 60 mg. the compounds of invention to adult males. 0135 Weekly assessments of depression or mental well With Compounds of Invention: With Placebo: being and Sexual function were done via widely accepted indices: to quantify depression and generalized anxiety Initial After Initial After disorders, the Hospital Anxiety and Depression Scale Participant Value Two-Weeks Value Two-Weeks (HAD); and for Sexual desire, psychological arousal, and 15 23.0, 8.0 18.0, 9.0 16.0, 9.0 16.0, 8.0 overall Sexual outlook, the Arizona Sexual Experience Index Average 13.5, 10.9 9.1, 10.4 11.8, 10.5 12.3, 10.8 (SEX). The HAD value precedes the ASEX one, and these are separated from each other by a comma. 0.136 Phase One lasted 14 days, during which partici Two-Sample Paired Sign Test - This is one of the stronger or more reli able statistical tests when significance is detected. The question is whether pants took daily doses of the invention or a placebo. ASSig INVENTION affects feelings of well-being or sexual function. The Sign nation of the invention or placebo to male participants was Test is used to statistically ask “how often compounds of invention impact done randomly. Immediately prior to the 14 days comprising feelings of well being and/or sexual function'. Results are as follows: HAD (with invention), p < 0.003, Very Significant Phase One, an initial physical examination and blood analy ASEX (with invention, p < 0.727, Not Significant sis were done. At that time, each male filled out HAD and HAD (with placebo), p < 0.688, Not Significant SEX forms to assess mental well being and Sexual function, ASEX (with placebo), p < 1.310, Not Significant was checked for Sitting and Standing blood pressure and pulse, and gave the blood Sample needed for the biochemical 0.139. These data indicate that the compounds of the analyses. invention have significant positive effect on depression or mood. Fourteen (14) of the fifteen (15) participants properly 0.137 Phase Two consisted of a seven-day period imme completed the Study and only data for these individuals were diately after Phase One, during which neither invention nor used for analysis. HAD scores exceeding 20.0 denote clini placebo was taken. During Phase Two, physical examination cal depression, but lower ones can also be associated with dispirited mood. Only two males entered the trial with HAD and laboratory analyses were again done. In Phase Three values exceeding the clinical minimum (21.0 and 23.0). which lasted 14 days, participants again took either inven Still, twelve (12) of fourteen (14) subjects showed bettered tion or a placebo. ASSignation of the invention or placebo mood, improved feelings of well being or lessened depres was done according to the Sort of capsule taken during Phase Sion after taking compounds of the invention. Decreases in One. If a participant took a compound of the invention in HAD Scores over the two-week timespan were as much as Phase One, then placebo was administered during Phase 15 and averaged 5.2. The two clinically depressed Subjects Three, and vice versa. Immediately after finishing Phase showed decreases in HAD values of 5.0 and 15.0. The Three, a physical examination and laboratory analyses were average HAD score went from 13.5 at the onset of the study again done. After completing Phase Three, each participant to 9.1 after two weeks of taking compounds of the invention, was asked prepared questions as well as Solicited for any very significant Statistically. Participants showed no Statis comments and impressions concerning invention use. tically detectable changes while taking placebo. 0140. After taking compounds of the invention for two 0138 Table 2 is a tabular summary of a presently pre weeks, five (5) of fourteen (14) participants had lessened ferred embodiment of results that may be observed in ASEX Values, indicating improved Sexual response or leSS Example 2. ened sexual anxiety, while only two (2) of fourteen (14) males showed the same after two weeks on placebo. Statis TABLE 2 tical Significance was not found for the ASEX changes, but Such could be attributed to a Small Sample size. Sexual HAD and ASEX summaries for administration of benefits of the compounds of the invention were also obvi the compounds of invention to adult males. ated in the trial through the exit interviews given to all participants. While taking these, a majority of males With Compounds of Invention: With Placebo: reported morning erections of the penis greater in Size, Initial After Initial After duration or frequency than usually experienced when not Participant Value Two-Weeks Value Two-Weeks taking the compound. Also, comments by the majority 1. 9.0, 12.0 8.0, 11.0 8.0, 10.0 8.0, 13.0 centered about feeling “like a teenager'(direct quote) in 2 16.0, 13.0 14.0, 12.0 16.0, 13.0 16.0, 15.0 terms of energy, Sexual and otherwise. It should be noted 3 15.0, 10.0 9.0, 11.0 15.0, 14.0 17.0, 14.0 that personal Situations were complicated by unwilling or 4 12.0, 9.0 7.0, 10.0 9.0, 10.0 9.0, 12.0 lacking sex partners. Twelve (12) of fifteen (15) participants 5 14.0, 10.0 10.0, 10.0 12.0, 10.0 14.0, 10.0 also expressed an unsolicited desire to continue using the 6 Participant not reliable - Data incomplete and deleted from study compounds of the invention. They stated a belief that the 7 12.0, 9.0 7.0, 9.0 9.0, 10.0 8.0, 9.0 compounds of the invention could prove useful to them in a 8 12.0, 11.0 8.0, 11.0 12.0, 11.0 12.0, 10.0 Sexual context. 9 12.0, 15.0 12.0, 10.0 7.0, 10.0 7.0, 10.0 EXAMPLE 3 1O 4.0, 15.0 0.0, 13.0 9.0, 12.0 12.0, 12.0 11 21.0, 10.0 6.0, 10.0 9.0, 12.0 12.0, 12.0 Compounds of the Invention as an Antidepressant 12 13.0, 9.0 2.0, 9.0 13.0, 9.0 13.0, 9.0 in Human Females 13 14.0, 12.0 13.0, 10.0 12.0, 10.0 12.0, 10.0 14 12.0, 10.0 13.0, 10.0 18.0, 7.0 16.0, 7.0 0.141. This example further relates to a method for less ening depression and otherwise bettering mood or feelings US 2005/0250772 A1 Nov. 10, 2005

of well-being, said method comprising the administration to EXAMPLE 4 human females of an effective amount of the compounds of the invention, defined above. Compounds of the invention Compounds of the Invention at Concentrations were used to treat eight females with clinical depression, Suitable for Human Therapies from Plants which for three females had been ongoing for at least one Harvested and Processed in Unique Fashion year. Participants took the same dose of compounds of the 0145 Example 4 relates to a method for obtaining com invention as in Example 2, 15-mg each day. A HAD was pounds of the invention at concentrations Suitable for human administered to participants prior to beginning daily doses. therapies from plants grown to an immature Stage of growth. Each participant was interviewed every two weeks to check “Concentrations Suitable for human therapies' means that for adverse side effects and for comments on use of the compounds of the invention in ten (10) grams or less of dried compounds of the invention. A HAD Index was again plant material make up a daily dose (e.g., fifteen (15) mg administered upon completion of the Six-week trial. compounds of the invention as 6-methoxy-2-benzoxazoli none; however, general therapeutically effective daily dos 0142 Referring Table 3, one presently preferred embodi ages of compounds of the invention may be between about ment of the results of the study are tabulated. five (5) mcg to about Sixty (60) mg). Said dosage may include either the novel compound as it naturally occurs or TABLE 3 Synthetically, or a combination of both natural and Synthetic novel compounds of the present invention. HAD summary for clinically-depressed females taking compounds of the invention for six weeks. 0146 Specific harvesting and drying conditions are Specified herein, as are analytical parameters for determin Initial After ing crop quality. By “specific harvesting and drying condi Participant' Value Six-Weeks tions”, it is meant that compounds of the invention are 1. 23.O 18.0 obtained from plants via circumstances differing from the 2 21.0 8.0 3 21.0 21.0 usual manner in which the plants are handled for the 4 24.0 14.O terminal product. 5 22.0 12.O 6 21.0 13.0 0147 AS an example, corn, Zea mayS, is typically grown 7 23.O 7.0 to its adult or matured States for its Seed-laden cob. At the 8 2O.O 22.0 immature growth Stage at which compounds of the invention Average 21.9 14.4 occur, corn plants have a biomass that portends harvest of a "Participants 1-4 initially Weeks 1-2 were given compounds of inven Substantial amount of leaf material containing the com tion under guise of its being a vitamin/mineral mixture. pounds of the invention at concentrations Suitable for human Two-Sample Paired Sign Test - This is one of the stronger or more reli therapies. Hence, dried corn leaves from immature plants able statistical tests when significance is detected. The question is whether compounds of invention affect depression or feelings of well-being. The become appropriate for the human therapies elucidated Sign-Test is used to statistically ask “how often compounds of invention herein, or the dried leaves are a resource for the concentra positively impact depression or feelings of well being. Results are as fol tion, extraction and purification of the compounds of the lows: invention. There are other monocotyledonous plants with HAD, p < 0.0313, Significant the natural production of compounds of the invention at concentrations Suitable for human therapies. 0143. These data indicate that the compounds of the 0.148 are flowering plants whose invention significantly leSSen depression. Initial HADScores embryo has only one cotyledon, or Seed leaf. The mono exceeded 20 for all subjects. All females were clinically cotyledons are a monophyletic group (i.e., derived from a depressed, and their HAD scores averaged 21.9. Six (6) of Single ancestral line) encompassing, but not limited to the eight (8) participants showed responses to compounds of the following botanical families: (true grasses, alter invention in which HAD values decreased 5-16 points, an nately referred to as Gramineae), Acoraceae (Sweet flag), average decrease of 10.5 over the Six-week timespan. The Agavaceae (yuccas), Alismataceae (water plantains), Ane overall average HAD score decreased from 21.9 (clinically marrhenaceae (Zhi mu), Anthericaceae (slender grass lily), depressed) at trial onset to 14.4 (not clinically depressed) Araceae (arum lilies), Asparagaceae (asparagus), Behni after Six weeks of use, with two participants ending with aceae (paleotropical lianas), Bromeliaceae (bromeliads, pineapple), Butomaceae (flowering rushes), Calamoideae HAD values of eight (8) and seven (7). There were only (palms or palm trees), Colchicaceae (milkmaids), Commeli eight females in the trial, but decreases in HAD Scores were naceae (blue gingers), Cymodoceaceae (shoalweed, Surf still statistically significant (p<0.031). The antidepressant grass, Widgeonweed), Cyperaceae (cotton grasses, Spike properties of the compounds of the invention are obviated. rushes, Sedges), Dioscoreaceae (yams), Eriocaulaceae (hat pins, pipewort), Flagellariaceae (false rattan), Hydrochari 0144. Both excess weight and Substance abuse are char taceae (waterweed), Iridaceae (iris), Juncaceae (rushes, acterized by either primary or Secondary depression. Since wood-rush), Juncaginaceae (arrow-weeds, arrow-grasses), Such psychological factors affecting exceSS weight and Sub Lanariaceae (Soapwort, latherwort, Scourwort), Lemnaceae stance abuse must be treated along with the physiological (duckweeds), Liliaceae (lilies, onions, blubells), Lowiaceae ones for therapies to be effective in the long term, the (orchidantha), Marantaceae (arrowroot, maranta, prayer compounds of the invention comprise adjunctive treatments plant), Mayacaceae (bog-moss, mayaca), Musaceae for achieving weight loSS or reducing the risk of relapse in (banana), Nolinaceae (beargrass), Orchidaceae (orchids), perSons with Substance abuse or addiction problems. Philydraceae (frogsmouth), Phormaliaceae (flax lily), Posi US 2005/0250772 A1 Nov. 10, 2005 doniaceae (posidonias), Potamogetonaceae (pondweeds), dried (96% dry weight) corn leaves with suitable levels of Rapateaceae (rapatea), Restionaceae (olifantsriet, rekoala compounds of the invention. Analyses via mass Spectros grass, Sprucecones), Ruppiaceae (large-fruit tassel, Widgeon copy showed that Substantive amounts of the compounds of grass), Sparganiaceae (bur-reeds), Themidaceae (Ookow, the invention were in dried corn leaves after five (5) weeks Bridges triteleia), Thurniaceae (prionium, thurnia), of growing time. Five (5) random Samples of dried corn Typhaceae (bulrushes), Xyridaceae (yellow-eyed grass), leaves were obtained for analysis. For each dried corn leaf Zingiberaceae (ginger), and Zosteraceae (eel grass). It is Sample, a one-gram portion was homogenized in 10-ml contemplated that compounds of the invention may be distilled water, incubated at 25 C. for one (1) hour, boiled derived isolated, harvested and/or extracted from plants for thirty (30) min, and then centrifuged for ten (10) min at selected from any of the above identified or other mono 3600 rpm. The resulting supernatant was extracted three (3) cotyledonous plant families or plant orders. times with ten (10) ml reagent-grade dichloromethane per 014.9 Preferred embodiments of the compounds of the extraction. The three (3) extracts were combined and invention may be Selected from families of Cyperaceae and allowed to air dry, after which the dried residue was stored Poaceae (alternately referred to as Graminae). Species in a tightly stoppered glass tube. therein include the cereal grasses (e.g., Sorghum, corn or 0152 The dried residue was analyzed for 6-MBOA by maize, wheat, rye, rice, barley and oats); Species related to gas-chromatographic mass spectroScopy. A Dupont Model the cereal grasses like gamagrass (Tripsacum species), wild DP102 device with an integrator and a SP2250 GC column rice (Zizania aquatica, Z. texana) and teosinte (Zea diplo isothermal at 200° C. (Dupont, Wilmington, Del.) was used. perennis, Z. luxurians, wild Z. mayS, Z. nicaraguensis, Z. A standard curve for 6-MBOA was obtained using 0.06-, perennis); Sugarcane (Saccharum species); (includ 0.60- and 1.20-mu.g. injections of pure, synthetic 6-MBOA ing but not limited to Species of Bambusa, BaShania, Cepha (Sigma, Saint Louis, Mo.) in methanol Solution and was loStachyum, Dendrocalamus, Dinochloa, Fargesia, Gigan reproducible at a five percent (5%) level. tochloa, Kinabaluchloa, Melocalamus, NaStuS, Phyllostchys, SchizoStacyum, Sinarundinaria, Thyrsos 0153 6-MBOA occurred in dried corn leaves at levels tachys, Vietnamosasa, and Yushania); and pasture grasses Suitable for human consumption. Samples averaged ten (10) (including but not limited to annual ryegrass, bent grass, mg/g 6-MBOA, with individual Samples assaying as fol Bermudagrass, bluegrass, kleingrass, orchard grass, rib lows: eight (8), nine (9), ten (10), ten (10), and twelve (12) grass, Sudangrass, timothy grass and fescue), along with mg/g 6-MBOA, respectively. At Such concentrations, leSS wild grasses (including but not limited to alkaligrass, than two (2) grams dried corn leaves are needed to make up bluestem grass, buffalograss, dallisgrass, hairgrass, Indian a daily human dose. This makes corn leaves, as uniquely grass, lovegrass, meadow grass, napiergrass, pampas grass, grown, harvested and dried herein, a Suitable Source of pangolagrass, panicgrass, rabbitsfoot grass, Slender wild compounds of the invention. For comparison purposes, oats, Switchgrass, toothbrush grass, wheatgrass); millet leaves from plants grown more than eight (8) weeks were (including but not limited to plants in the genera Panicum, analyzed for 6-MBOA. Virtually none was present. Setaria, Echinochloa, Pennesetum and Eleusine); Job’s tears 0154 Previous work by the inventors indicated that (Coix lachryma-jobi, Coix aquatica); and the barley-like higher levels of the compounds of the invention are asso grasses (including but not necessarily limited to species of ciated with multiple biochemical parameters that indicate Hordeum, Setaria, Astrebla, Isolepis, Luzula and Hyparrhe crop quality or adequacy with respect to the compounds of nia). the invention. In those plants containing compounds of the 0150 Growing corn to obtain the compounds of the invention at concentrations Suitable for human use total invention is initially done in a conventional fashion, but phenols are at concentrations greater than 17.0 mg/gm (dry Seeds are planted more densely than is the case for conven weight) but combined amounts of 4-hydroxycinnamic acid tional crops because of the Smaller Size of plants at harvest. and 4-hydroxy-3-methoxycinnamic acid total no more than Harvest time is done while plants are immature. For corn, 1.5 mg/gm (dry weight), as determined through chromatog this immature plant harvesting may happen when plants are raphy. For invention, the above mentioned parameters for no more than about thirty (30) to about forty-five (45) total phenols and combined amounts of 4-hydroxycinnamic centimeters tall, about five weeks after planting. Preferably, acid and 4-hydroxy-3-methoxycinnamic acid are instituted embodiments of the invention may also utilize harvesting here as elements of the invention as it pertains to plants. For immature corn plants that are between about forty-five (45) the corn leaf Samples of Example 2, total phenols averaged centimeters to about 122 centimeters in height, and between 19.1 mg/gm and the cumulative total for cinnamic acids about five weeks to about eight weeks after planting are averaged 0.9 mg/gm. Sought and preferably less than ten weeks. As a general reference, mature corn plants are typically more than 180 EXAMPLE 5 centimeters in height and are grown for about four (4) to about five (5) months after planting. For corn, harvesting is Compounds of the Invention from Parts of Animals preferably done by cutting plants at 3-4 centimeters above 0155 The main food of deer and elk (Cervidae) for most the ground. Severed plants may be gathered and may be of the year is browse, the growing tips of low-growing, dried at temperatures held at about 40° Celsius (C) to about Woody plants. However, casting of hard antlers from the 45 C. Empirical studies showed that this temperature range previous year coincides with a Spring flush in natural pas helps maximize conversion of the precursors of compounds turage and an accompanying shift to a diet of grasses, chiefly of the invention to the active molecules. Sprouting and immature ones. Such grasses are at develop 0151. In a trial plot in Southern Illinois, approximately mental stages in which 6-MBOA and related compounds are 38,000 corn plants yielded 137 kilograms (300 pounds) of most prevalent. US 2005/0250772 A1 Nov. 10, 2005

0156 After casting, new antlers begin their development. those characterized by high fat contents. Obtaining com These growing antlers are nourished by blood vessels from pounds of the invention from body parts other than the a covering of Skin, called Velvet. An antler grows from the antlers of elk and deer and from animals other than elk or tip with tissue laid down as the tip advances. A velvet antler deer are not precluded from invention. tip has a Soft cartilaginous internal Structure and high fat content, contrasting the rest of the antler with its OSSified EXAMPLE 6 cartilage and little fat. Alternative Embodiments of the Novel Compounds O157 Air-dried and freeze-dried velvet antler samples were obtained. These came from commercially farmed 0163 Any number of alternative embodiments of precur Canadian Wapiti and New Zealand red deer, both subspecies Sors of phenolic compounds of the present invention may be of elk, Cervus elaphus. All animals had been maintained on contemplated as falling within the Spirt and Scope of the grassy pasturage. The Samples were from Velvet antlers that present invention. In particular, Formula V, below, is a had been growing fifty-five (55) to sixty-five (65) days, and general chemical formula depicting a generic representation included both tips, defined as the region five (5) cm or less of alternative preferred embodiments of precursors of phe in length starting at the apex, as well as other, more matured nolic compounds of the present invention. parts of the antler. 0164. Formula V-A compound according to the for 0158 For each sample of dried antler, a one-gram portion mula: was homogenized in ten (10)-ml distilled water, incubated at R 25 C. for one (1) hour, boiled for thirty (30) min, and then centrifuged for ten (10) min at 3600 rpm. The resulting Supernatant was extracted three (3) times with ten (10) ml reagent-grade dichloromethane per extraction. The three (3) N O extracts were combined and allowed to air dry, after which R3 the dried residue was Stored in a tightly Stoppered glass tube. 0159. The dried residue was analyzed for 6-MBOA by 01.65 Wherein “R” is selected from the group con gas-chromatographic mass spectroScopy. A Dupont Model sisting of H and OCH; DP102 device with an integrator and a SP2250 GC column isothermal at 200° C. (Dupont, Wilmington, Del.) was used. 0166 Wherein “R” is selected from the group con A standard curve for 6-MBOA was obtained using 0.06-, Sisting of H and Glucose (as a glucoside) 0.60- and 1.20-mcg injections ofpure, synthetic 6-MBOA 0167. Wherein “R” is selected from the group con (Sigma, Saint Louis, Mo.) in methanol Solution and was sisting of H, OH, and OCH; reproducible at a five percent (5%) level. 0168 or pharmaceutically acceptable salts thereof. 0160 Referring to Table 4, the observations of Example 5 may be summarized as follows: 0169. As shown, such embodiments of novel compounds of the present invention may include benzoxazinoids-cyclic TABLE 4 hydroxyamic acids, lactams and corresponding glucosides. As contemplated herein, Substitution at the “R” position 6-MBOA in Dried Velvet Antler from Elk, Cervus elaphus. may be accomplished with a member Selected from the Drying 6-MBOA group consisting of H and OCH. Substitution at the “R” Animal Origin Tip or Other Method (mg/g dry weight) position may be accomplished with a member Selected from the group consisting of H and glucose (as a glucoside). Wapiti Canada Tip Air 2.5 Wapiti Canada Tip Air 2.8 Substitution at the “R” position may be accomplished with Wapiti Canada Other Air O.3 a member Selected from the group consisting of H, OH, and Red Deer New Tip Freeze 1.9 OCH. Zealand Red Deer New Other Freeze 0.5 0170 Referring now compounds 8-17 below, a series of Zealand chemical formulae, according to the generic representation shown in Formula V, illustrate chemical structures for rep resentative compounds of further embodiments of novel 0.161 These data indicate 6-MBOA was present in all tip compounds of the present invention. Samples, with little or none present in those from the more matured parts of the antler. Notably, amounts of 6-MBOA O171 8 2.4-dihvdroXV-1,4-b4-dunydroxy-1,4-benzoxazin-3-one in-3 from Velvet antler tips exceeded those typically found in (DIBOA) grasses less than a week after Sprouting, the Stage of growth with the most 6-MBOA. These results show that ingested 6-MBOA is accumulated or concentrated in velvet antler tips, and as Such represent a means for obtaining the compounds of the invention in concentrations Suitable for H C human use. O 0162 Many animals eat grasses and other monocotyle OH donous plants. Such animals may also be accumulating compounds of the invention in body parts, most likely in US 2005/0250772 A1 Nov. 10, 2005 15

0172 9. 2,4-dihydroxy-1,4-benzoxazin-3-one-gluco 0177 14. 2-hydroxy-7-methoxy-1,4-benzoxazin-3- side (DIBOA-Glc) one (HMBOA)

H H3CO

O O H OH

0173 10. 2,4-dihydroxy-7-methoxy-1,4-benzoxazin 0.178 15. 2-hydroxy-7-methoxy-1,4-benzoxazin-3- 3-one (DIMBOA) one-glucoside (HMBOA-Glc)

HCO HCO DC -Glc

O O H OH

0179 16. 2-hydroxy-4,7-dimethoxy-1,4-benzoxazin 0.174 11. 2,4-dihydroxy-7-methoxy-1,4-benzoxazin 3-one-glucoside (DIMBOA-Glc) 3-one (HDMBOA) HCO C H3CO C O O OCH3 OH

0180 17. 2-hydroxy-4,7-dimethoxy-1,4-benzoxazin 0175 12. 2-hydroxy-1,4-benzoxazin-3-one (HBOA) 3-one-glucoside (HDMBOA-Glc) H3CO DC -Glc H C O O OCH3

H 0181 Specifically, compound 8 illustrates a chemical formula for 2,4-dihydroxy-1,4-benzoxazin-3-one (DIBOA). DIBOA may also have a glucose molecule to form a glucoside (also referred to as a glycoside), DIBOA-Glc, 0176 13. 2-hydroxy-1,4-benzoxazin-3-one-glucoside which is shown compound 9. As illustrated, compound 10 (HBOA-Glc) depicts a chemical formula for 2,4-dihydroxy-7-methoxy-1, 4-benzoxazin-3-one (DIMBOA). DIMBOA may also exist in combination with glucose molecule to form a glycoside H compound (DIMBOA-Glc), which is shown as compound 11.

O 0182 Compound 12 illustrates a chemical formula for 2-hydroxy-1,4-benzoxazin-3-one (HBOA). A glycoside may H also form between HBOA and a glucose molecule (HBOA Glc) and is shown as compound 13. Compound 14 depicts a chemical formula for 2-hydroxy-7-methoxy-1,4-benzox US 2005/0250772 A1 Nov. 10, 2005 azin-3-one (HMBOA). HMBOA may also contain a glucose are found in bamboo are quite extant immediately prior to molecule to form HMBOA-Glc, which is depicted as com and during the breeding Season. During Such times, bamboo pound 15. Additionally, compound 16, illustrates a chemical shoots and younger leaves, those States in which the com formula for 2-hydroxy-4,7-dimethoxy-1,4-benzoxazin-3- pounds of the invention are particularly plentiful, may be one (HDMBOA). Wherein, compound 17 illustrates an preferentially eaten by giant pandas. Meanwhile, the glucoside formed between HDMBOA and glucose (HDM matured or older bamboo leaves So prevalent at other times BOA-Glc). In the foregoing examples, glucose molecules have little or none of compounds contemplated by the may be bonded to a respective aglycone (i.e., non-Sugar) presently preferred embodiments of the present invention. compound (e.g., DIBOA, DIMBOA, HBOA, HDMBOA, 0187 Adult giant pandas may consume between about 6-MBOA) to form a glycoside. twelve (12) to about fifteen (15) kilograms of food per day 0183 AS appreciated, glucose molecules typically are in when feeding on matured or older bamboo leaves and Stems. the form of a pyranose (i.e., cyclic 6-carbon ring), which However, when feeding in nature on bamboo shoots, which may be referred to as a glucopyranose. Glucopyranose have the compounds in accordance with presently preferred compounds usually bond with the aglycone portion as an embodiments of the present invention, giant pandas may hemiacetal. Configurations of glycoside compounds in yet consume up to thirty-eight (38) kilograms per day. This may other presently preferred embodiments of the present inven account for about forty percent (40%) of their average body tion may be found in the (2R)-configuration. It is intended, weight. however, that other forms of glucose and configurations of glycosides are contemplated to be within the Spirt and Scope 0188 Historically, the compounds of the present inven of the novel compounds of the present invention. The tion may almost always be lacking in Zoo diets fed to giant presently preferred embodiments of compounds of the pandas. Unlike the diet of wild pandas, captive animals are present invention, as shown in Example 6, are intended typically fed only older bamboo, with only minuscule therefore to be viewed as exemplary of the principles of the amounts of the active compounds of the present invention. present invention, and not as restrictive to any particular Captive pandas may also be fed rice gruel, carrots, apples, formula, Structure or method for implementing and/or prac ground corn and Sweet potatoes. ticing the present invention. 0189 It has been previously demonstrated that the com pounds in accordance with presently preferred embodiments EXAMPLE 7 of the present invention may be passed from giant panda mothers to offspring during gestation and/or lactation (Nel Novel Compounds of the Invention in Preserving Son, “Maternal diet influences reproductive development in and/or Augmenting Innate Immune Defenses in male prairie Vole offspring.” Physiology of Behavior Giant Pandas 50: 1063-1066). Accordingly, female giant pandas who may 0184 The endangered giant panda (Ailuropoda melano be fed the active compounds of the present invention may be leuca) is a bear-like mammal native to the high bamboo and examined from the perspective of preweaning baby panda pine forests of central China. The total number of giant Survival. pandas in the wild is now about 1,500, with another 140 animals housed in Zoos and research centers. These numbers 0190. In this example, bamboo with a relative abundance may place giant pandas as one of the most endangered of the compounds comprising the invention were fed to Species on Earth. One reason that giant panda populations animals at the Giant Panda Breeding Center, Wolong Nature may be at endangered levels is due to high infant mortality Reserve, Sichuan Province, China. There are more than forty rates. Babies born in captive panda populations may have (40) adult pandas at the Center, which accounts for thirty less than a fifty percent (50%) chance of survival to wean percent (30%) of the total number of captive pandas in the ing. Of forty-two (42) giant pandas produced in captivity world. between 1980 and 1997, only twenty-three (23) have Sur 0191 Levels of the active compounds of the present Vived. It has been documented that giant panda babies invention may be determined for shoots, younger leaves usually die because of infectious disease and/or an immature emerging from the Shoots and matured, older leaves of immune System. Fargesia Spathacea, the most usual bamboo Species fed at 0185. Wild giant pandas may primarily eat bamboo. the Wolong Giant Panda Breeding Center. Three (3) samples Bamboo may be categorized as a monocotyledonous plant. of each developmental State of bamboo were taken and dried Moreover, giant pandas occasionally eat flowers, Vines, at 40-45 C. For each dried bamboo leaf or shoot sample, grasses, pine bark, honey, insects and rodents. Certain approximately a one-gram portion was homogenized in ten bamboo Species may be widespread at the high altitudes that (10)-mL distilled water, incubated at 25 C. for about one (1) pandas may now inhabit. Species of bamboo may include, hour, boiled for about thirty (30) min and then centrifuged for example and not by way of limitation, Fargesia Spatha for about ten (10) minutes at approximately 3600 rpm. The cea, Sinarundinaria chungii, S. nitida, and S. fangiana. All resulting Supernatant was extracted three (3) times with ten may invariably contain compounds in accordance with pres (10) mL reagent-grade dichloromethane per extraction. The ently preferred embodiments of the present invention in the three (3) extracts were then combined and allowed to air dry, shoots and otherwise young leaves. There may be virtually after which the dried residue may be stored in a tightly none of the compounds of the present invention, however, Stoppered glass tube. occurring in the older life history Stages of these bamboo 0.192 The dried residue was analyzed for 6-methoxy-2- plants. benzoxazolinone (6-MBOA) by gas-chromatographic mass 0186. In particular, compounds in accordance with pres spectroscopy. A Dupont Model DP 102 device with an ently preferred embodiments of the present invention that integrator and a SP2250 GC column isothermal at 200 C. US 2005/0250772 A1 Nov. 10, 2005

(Dupont, Wilmington, Del.) was used. A standard curve for ronment for a desired period of time Sufficient to Suppress 6-MBOA was obtained using 0.06-, 0.60- and 1.20-ug innate immune responses, then Subjecting the animals to an injections of pure, synthetic 6-MBOA (Sigma, Saint Louis, agent known to precipitate an immune response (i.e., immu Mo.) in methanol solution and was reproducible at a five nogenic or antigenic agent). Typically, the immune response percent (5%) level. may be measured by the amount of localized Swelling (i.e., inflammation) that may occur near the administration site of 0193 6-MBOA contents (in parts per million) for the the immunogenic agent. The effects of a particular immu nine (9) Fargesia Spathacea Samples were, as follows: (1) nogenic agent on the immune System of an animal may be Shoots=56, 88, 91; (2) Younger leaves=23, 27, 38; (3) examined by comparing the degree of Swelling in control Mature, Old Leaves=3, 3, 5. and experimental groups of the animals. 0194 During the 2003 Breeding Season-approximately March though May-captive pandas may receive a diet 0199 Male albino inbred strain mice, Balb/c (Charles consisting chiefly of bamboo shoots and young leaves in the River, Inc.), at about eight (8) weeks of age were used in the month prior to and also during the March through May present exemplary Study. AS appreciated by those skilled in reproductive period. Average consumption may be on the the art, Balb/c mice are often used in StreSS/injury testing range of about twenty-eight (28) kilograms of bamboo per because they may be more Susceptible to StreSS and injury day over this time span. Rice and vegetable gruel may also compared to other Strains of animals and, importantly, be fed to the pandas. Older bamboo leaves are preferably immune System changes are more easily observed in these precluded, as much as possible, in contrast to past breeding animals. Seasons when these encompassed most of the diet. 0200. It is anticipated that the mice that are stressed, but also exposed to immature corn leaves containing 6-MBOA 0195 Prior to concerted efforts to feed bamboo at devel or related compounds of the present invention (i.e., stressed/ opmental States characterized by notable amounts of the 6-MBOA mice) may have the ability to mount an immune compounds of the present invention, only ten percent (10%) response to a known immunogenic agent. In contrast, those of captive male pandas at the Breeding Center reproduced in mice that may be stressed, but not exposed to immature corn a natural way, less than thirty percent (30%) of females gave leaves containing 6-MBOA or related compounds of the birth and more than fifty percent (50%) of the panda babies present invention (i.e., Stressed/non-6-MBOA mice) may died prior to weaning. Bamboo fed during previous repro have little or no ability to mount an immune response to a duction Seasons consisted chiefly of mature older plants. known immunogenic agent. Likewise, additional control Indeed, older plant material may have been readily and groups may utilize mice that are not stressed, and are not cheaply obtainable in large amounts, So little or no younger exposed to immature corn leaves containing 6-MBOA or material had been offered on a concerted basis let alone in related compounds of the present invention (i.e., non quantity. stressed/non-6-MBOA mice) and mice that are stressed, but 0.196 Based on the findings and research conducted at the are not exposed to immature corn leaves containing Giant Panda Breeding Center, with bamboo containing the 6-MBOA or related compounds of the present invention compounds in accordance with presently preferred embodi (i.e., stressed/non-6-MBOA mice). ments of the present invention making up a large part of the 0201 In the present study, twenty (20) Balb/c mice were diet, the birthrate increased about three-fold (3x), with evenly divided into four (4) groups. These groups comprised almost all captive females achieving pregnancy. Baby Sur the following characteristics: Group 1 included a control Vival past weaning, which had previously entailed a high group of non-stressed/non-6-MBOA mice; Group 2 included mortality due to infectious disease, became almost total a group of stressed/non-6-MBOA mice; Group 3 included a Survival. Indeed, a record number of young were pro group of non-stressed/6-MBOA mice and Group 4 included duced-nineteen (19) offspring-during this example in the a group of stressed/6-MBOA mice. 2003 Breeding Season at the Giant Panda Breeding Center. Of these offspring, two (2) had been stillborn and one (1) 0202 Beginning on day 3 prior (Day -3) to exposure to died quite soon after birth. Necropsy of the three (3) animals a StreSS procedure, immature corn leaves containing revealed nothing as to any problems of an infectious or 6-MBOA or related compounds of the present invention was pathogenic nature. The remaining offspring have continued administered to the mice in Groups 3 and 4 orally via feed. to thrive through January 2004. In Summary, unprecedented The mouse chow was chopped into small bits, followed by Survival of giant pandas may be observed following admin the addition of immature corn leaves containing 6-MBOA or istration of compounds of the present invention, in View of related compounds of the present invention at an appropriate the normally expected greater than fifty percent (50%) infant concentration (e.g., 40 mg/kg/mouse/day). The mouse chow mortality based on the outcome of previous reproductive and immature corn leaves containing 6-MBOA or related SCSOS. compounds of the present invention was re-blended with Some honey and water, and then dried. The mice in Groups EXAMPLE 8 1 and 2 had food prepared the same way, without the addition of immature corn leaves containing 6-MBOA or Novel Compounds of the Invention in Preserving related compounds of the present invention. All of the and/or Augmenting Innate Immune Defenses in Groups were fed in the same manner on day 2 (Day -2) and Mice day 1 (Day -1) prior to exposure to a stress procedure. 0197) Study Design 0203 On day Zero (Day 0), the mice in Groups 2 and 4 were stressed using a cold restraint StreSS procedure. AS 0198 A model for examining stress-induced changes in appreciated by those skilled in the art, the cold restraint animals may involve Subjecting animals to a stressful envi StreSS procedure is a well described model that may result in US 2005/0250772 A1 Nov. 10, 2005 a decreased immune response and greater Susceptibility to immune responsiveness to an immunogenic agent. More infection. In the cold restraint StreSS procedure, the mice specifically, the cellular immune response to DNFB was were placed in ventilated fifty (50) mL centrifuge tubes determined by Sensitizing the mice by topical application of where they were able to move forward and backward, but DNFB. As illustrated in FIG. 1, the ordinate (vertical) axis were unable to turn around. The mice were then be placed represents the difference in thickness (i.e., Swelling), mea in a refrigerator at 4°C. for about one (1) hour, then returned Sured in millimeters (mm), between the right ear pinna and to their cages. The cold restraint StreSS procedure lacked the contralateral ear pinna. The abscissa (horizontal) axis tissue injury, and most closely resembled human psycho represents a bar corresponding to each of the four mice logic StreSS and/or the shipping or other Stresses that various Groups identified in this study. animal Species may experience (e.g., confinement, lack of food, lack of water, psychological distress, environmental 0211. In one presently preferred embodiment of the changes, presence or absence of light, and the like) by present invention, a solution of 0.25% DNFB (Sigma) was analogy. prepared in a 4:1 concentration of acetone-mineral oil mix ture, and 10 microliters (ul ) was applied to the back of the 0204. The immune measure chosen was the delayed right ear pinna on Day +1 and Day +2. On Day +4, the ears hypersensitivity response to dinitrofluorobenzene (DNFB), were measured with a caliper to 10 mm. Swelling of the an immunogenic agent. DNFB may Sometimes be referred ear (higher measurement number) indicates an immune to as 1-fluor-2,4-dinitrobenzene, 2,4-dinitro-1-fluoroben response to the DNFB, whereas the greater the Swelling, the Zene, or Sanger reagent. The mice were exposed to DNFB greater magnitude of response. via administration to ear skin on Days +1 and +2. In normal animals, the challenged areas may Swell in response to the 0212. The results of the study showed a decrease in administration of DNFB, which may reflect a specific sen immune response in control Stressed mice, as compared with Sitization to this antigen. Such SWelling typically represents all other GroupS. Stressed mice exposed to immature corn inflammation and a measure of T-cell mediated immune leaves containing 6-MBOA or related compounds of the response to DNFB. Animals which are stressed or injured or present invention had normal responses to the DNFB, Sug Stressed in Some way have blunted responses, reflecting gesting that immature corn leaves containing 6-MBOA or immunosuppression. This response has also been associated related compounds of the present invention preserved cel with resistance to microbial infections. lular immune response in restraint Stressed mice. 0205. On day 1 (Day +1) and day 2 (Day +2) following EXAMPLE 9 the StreSS procedure, all mice in Groups 1-4 were exposed to DNFB via administration to skin on the right ear pinna (i.e., Novel Compounds of the Invention in Preserving ear skin). On Day +4, the thickness of the ear skin was and/or Augmenting Innate Immune Defenses in measured with caliperS on the exposed ear of each of the Mice-Dosage Range Study mice and the thickness of the ear Skin on a contralateral ear (i.e., opposite ear non-exposed to DNFB). The ear skin 0213 Study Design thickness value of the non-exposed skin was Subtracted from 0214) Another study may be performed to evaluate the the ear skin thickness value of the DNFB exposed skin to effect on cell mediated immune response from immature quantify the level of immune response to DNFB. corn leaves containing 6-MBOA or related compounds of 0206. The study protocol, presented hereinabove, may be the present invention administered to Stressed mice at a Summarized in Table 5, as follows: plurality of concentrations. The Study protocol was similar to the one exercised in Example 8, with mice exposed to immature corn leaves containing 6-MBOA or related com pounds of the present invention at dosage ranges of 40 Day Action mg/kg, 25 mg/kg and 5 mg/kg 6-MBOA (6-MBOA Dose # -3 new mouse chow with/without 6-MBOA begun 1, 2 and 3, respectively). The thickness of DNFB challenged -2 new mouse chow with/without 6-MBOA continued ears was measured on day +4. -1 new mouse chow with/without 6-MBOA continued O Groups 2 and 4 exposed to cold stress procedure 0215 Male albino inbred strain mice, Balb/c (Charles +1 All mice exposed to DNFB on right ear skin River, Inc.), at about eight (8) weeks of age were used in this +2 All mice exposed to DNFB on right ear skin example. Balb/c mice may often be used in StreSS/injury +4 All mice ears measured with calipers animals because they may be more Susceptible to StreSS and injury compared to other Strains of animals. Those skilled in the art will appreciate that immune System changes are more 0207 Statistical Analysis easily observed in these animals. 0208. The ear skin Swelling results were analyzed using 0216. It is anticipated that the mice that are stressed, but analysis of variance (ANOVA) and Fisher's Protected Least also exposed to immature corn leaves containing 6-MBOA Square Difference (PLSD) tests. or related compounds of the present invention (i.e., stressed/ 6-MBOA mice) may have the ability to mount an immune 0209 Results response to a known immunogenic agent. In contrast, those 0210 Referring generally now to FIG. 1, the stressed mice that may be stressed, but not exposed to immature corn mice exhibited a depressed immune response compared to leaves containing 6-MBOA or related compounds of the the unstressed mice. Moreover, the Stressed mice that were present invention (i.e., Stressed/non-6-MBOA mice) may fed immature corn leaves containing 6-MBOA or related have little or no ability to mount an immune response to a compounds of the present invention showed normal levels of known immunogenic agent. Likewise, additional control US 2005/0250772 A1 Nov. 10, 2005 groups may utilize mice that are not stressed, and are not DNFB via administration to skin on the right ear pinna (i.e., exposed to immature corn leaves containing 6-MBOA or ear skin). On Day +4, the thickness of the ear skin was related compounds of the present invention (i.e., non measured with caliperS on the exposed ear of each of the stressed/non-6-MBOA mice) and mice that are stressed, but mice and the thickness of the ear Skin on a contralateral ear are not exposed to immature corn leaves containing (i.e., opposite ear non-exposed to DNFB). The ear skin 6-MBOA or related compounds of the present invention thickness value of the non-exposed skin was Subtracted from (i.e., stressed/non-6-MBOA mice). the ear skin thickness value of the DNFB exposed skin to 0217. Thirty-five (35) Balb/c mice were divided into six quantify the level of immune response to DNFB. (6) groups. These groups comprised the following charac 0222. The study protocol, presented hereinabove, may be teristics: Group 1 included a control group of non-stressed/ Summarized in Table 6, as follows: non-6-MBOA mice; Group 2 included a control group of stressed/non-6-MBOA mice; Group 3 included a group of non-stressed/6-MBOA mice; Group 4 included a group of stressed/6-MBOADose #1 mice; Group 5 included a group Day Action of stressed/6-MBOA Dose #2 mice and Group 6 included a -3 new mouse chow with/without 6-MBOA begun group of stressed/6-MBOA Dose #3 mice. -2 new mouse chow withfwithout 6-MBOA continued -1 new mouse chow withfwithout 6-MBOA continued 0218 Beginning on day 3 prior (Day -3) to exposure to O Groups 2 and 4-6 exposed to cold stress procedure a StreSS procedure, immature corn leaves containing +1 All mice exposed to DNFB on right ear skin 6-MBOA or related compounds of the present invention was +2 All mice exposed to DNFB on right ear skin administered to the mice in Groups 3-6 orally via feed. The +4 All mice ears measured with calipers mouse chow was chopped into small bits, followed by the addition of immature corn leaves containing 6-MBOA or related compounds of the present invention at an appropriate 0223 Statistical Analysis concentration of 6-MBOA (e.g., 40 mg/kg/mouse/day). The 0224. The ear skin Swelling and footpad Swelling results mouse chow and immature corn leaves containing 6-MBOA were analyzed using analysis of variance (ANOVA) and or related compounds of the present invention were re Fisher's Protected Least Square Difference (PLSD) tests. blended with some honey and water, and then dried. The mice in GroupS 1 and 2 had food prepared the same way, 0225 Results without adding immature corn leaves containing 6-MBOA 0226 Referring now to FIG. 2, the stressed mice exhib or related compounds of the present invention. All Groups ited a depressed immune response compared to the were fed in the same manner on day 2 (Day -2) and day 1 unstressed mice. Moreover, Stressed mice receiving imma (Day -1) prior to exposure to a stress procedure. ture corn leaves containing 6-MBOA or related compounds 0219. On day Zero (Day 0) the mice in Groups 2 and 4 of the present invention at concentrations of 40 mg/kg and were stressed using a cold restraint StreSS procedure. AS 25 mg/kg 6-MBOA had significantly greater immune appreciated by those skilled in the art, the cold restraint responses than animals given immature corn leaves contain StreSS procedure is a well described model that typically ing 6-MBOA or related compounds of the present invention results in a decreased immune response and greater Suscep at concentrations of 5 mg/kg 6-MBOA or no 6-MBOA. tibility to infection. In the cold restraint StreSS procedure, the mice were placed in ventilated fifty (50) mL centrifuge tubes EXAMPLE 10 where they were able to move forward and backward, but were unable to turn around. The mice may then be placed in Novel Compounds of the Invention in Preserving a refrigerator at a temperature of about 4 C. for one (1) and/or Augmenting Innate Immune Defenses in hour, then returned to their cages. The cold restraint StreSS Calves procedure lacked tissue injury, and most closely resembled human psychologic StreSS and/or the Shipping or other 0227 Study Design stresses that various animal species may experience (e.g., 0228. A model for examining stress-induced changes in confinement, lack of food, lack of water, psychological animals may involve Subjecting animals to a stressful envi distress, environmental changes, presence or absence of ronment for a desired period of time Sufficient to Suppress light, and the like) by analogy. innate immune responses, then Subjecting the animals to an 0220. The immune measure that was chosen was the agent known to precipitate an immune response. Typically, delayed hyperSensitivity response to dinitrofluorobenzene the immune response may be measured by the amount of (DNFB). The mice were exposed to DNFB via administra localized Swelling (i.e., inflammation) that may occur near tion to ear skin on Days +1 and +2. In normal animals, the the administration site of the agent. The effects of a particu challenged areas may Swell in response to the administration lar agent on the immune System of the animals may be of DNFB, which may reflect a specific sensitization to this examined by comparing the degree of Swelling in control antigen. Such Swelling typically represents inflammation and experimental groups of the animals. and a measure of T-cell mediated immune response to 0229. In the present study, immature corn leaves contain DNFB. Animals which are stressed or injured or stressed in ing 6-MBOA or related compounds of the present invention Some way have blunted responses, reflecting immunoSup were mixed with grain and fed to weaned calves who may pression. This response has also been associated with resis be subjected to a stressful challenge (e.g., halter breaking). tance to microbial infections. Evidence of biologic effects of Supplementation with imma 0221) On day 1 (Day +1) and day 2 (Day +2) following ture corn leaves containing 6-MBOA or related compounds the StreSS procedure, all mice in Groups 1-6 were exposed to of the present invention may be observed, Suggesting that US 2005/0250772 A1 Nov. 10, 2005 metabolism of 6-MBOA or related compounds of the present there was no significant difference in Starting weights invention occurred and may indicate preservation and/or between each of the Groups. There were no differences augmentation of immune response in Stressed calves. between Groups at any time point, whereas Supplementation 0230. In one presently preferred embodiment of the with immature corn leaves containing 6-MBOA or related present invention, the Study facility included a barn with compounds of the present invention did not inhibit weight fencing, a Squeeze chute, Scale and an arena were the calves gain of the calves. may be assembled for procedures. There was also a class 0238) Immune Response ASSay room utilized as a laboratory. 0239). The cell mediated immune response was assessed 0231. Thirty-two (32) calves were evenly divided into in all calves using DNFB sensitization. The calves were four (4) groups comprising eight (8) animals in each group. placed in the chute on Day 0, a Section of their left necks These four (4) groups comprised the following characteris shaved and 100 ul 4% DNFB in acetone and mineral oil tics: Group 1 included a control group of non-stressed/non applied to the Shaved area. On Day +1, this was repeated, 6-MBOA calves; Group 2 included a group of stressed/non using 400 ul DNFB on the same neck area. On Day +3, the 6-MBOA calves; Group 3 included a group of stressed/6- calves right ear was shaved and 60 ul DNFB was applied to MBOA Dose #1 calves and Group 4 included a group of the Shaved area. The Sensitization area of the neck and stressed/6-MBOA Dose #2 calves. normal neck skin was measured using a caliper. Twenty four (24) hours later, the challenged ear and the contra lateral ears 0232 The calves were mostly females (n=29), and also were both measured, and the neck skin was re-measured and included a steer (n=1) and bulls (n=2). The calves were the data recorded. The data are expressed differences weaned for a period of at least two weeks. StreSS was between normal and Sensitized or challenged skin. precipitated in the calves via roping. The calves had evi dence of ring Worm infection, but none of the calves 0240 Immature corn leaves containing 6-MBOA or exhibited an acute respiratory or other infection. The calves related compounds of the present invention was Successfully all appeared to have good appetites and readily consumed mixed with grain and feed to the calves who readily con the grain rations. Sumed the rations. 0233. The calves were housed outdoors in four separate 0241 Results corrals, with water and hay available ad libitum. The calves 0242. As observed in the mouse experiments referenced were fed Sweet mix grain, with and without Supplementation hereinabove, the immune response to a contact antigen Was of immature corn leaves containing 6-MBOA or related greater in calves exposed to immature corn leaves contain compounds of the present invention. Beginning on day 1 ing 6-MBOA or related compounds of the present invention prior (Day -1) to exposure to a stress procedure, the calves as compared to stressed controls. Referring now to FIG. 3, were fed once in the morning (8 am) on Day -1 and on Day immune response, measured both as degree of Swelling at 0 by spreading the grain ration over a shallow feed trough. the Sensitization site and at the challenged ear, was greater Beginning on Day +1 and continuing until the end of the in the calves exposed to immature corn leaves containing Study, the calves were fed grain twice daily at 8 a.m. and 5 6-MBOA or related compounds of the present invention, as p.m. The calves lined up at the trough, and were observed to compared to the Stressed controls. Immune response may be eat approximately equal amounts of the ration. Control measured in millimeters difference using a caliper measur groups (i.e., Groups 1 and 2) and groups Supplemented with ing device. AS illustrated, GroupS3 and 4 were able to mount immature corn leaves containing 6-MBOA or related com an immune response following exposure to DNFB. pounds of the present invention (i.e., Groups 3 and 4) all 0243 Referring to FIG. 4, immune response, measured received approximately ten (10) lbs of Sweet mix grain twice both as degree of Swelling at the Sensitization site and at the daily. All rations were typically consumed within about five challenged area of the neck, was greater in the calves (5) minutes. exposed to immature corn leaves containing 6-MBOA or 0234. The stress procedure for this study involved halter related compounds of the present invention, as compared to breaking. Halter breaking included a 1.5 hour tie up (i.e., the Stressed controls. AS illustrated, Groups 3 and 4 were roping) at a fence in the arena. During this roping procedure, able to mount an immune response following exposure to the calves were prevented from obtaining food or water. The DNFB. roping occurred on Day 0 and was immediately followed 0244. Discussion and Conclusions with sensitizing of the calves necks with DNFB. 0245. In conclusion, it is apparent that Supplementation 0235 Immature corn leaves containing 6-MBOA or with immature corn leaves containing 6-MBOA or related related compounds of the present invention may be weighed compounds of the present invention has positive effects in on a laboratory Scale, and the appropriate amount for each calves Subjected to StreSS challenge. This was a robust group of 8 animals was added to seven (7) lbs of Sweet mix observation, given the variability in the genetics and sizes of grain and mixed thoroughly by hand. The Sweet mix grain the calves, and the relatively poor condition of the Subjects was Somewhat Sticky and the immature corn leaves contain at the Start of the experiment. ing 6-MBOA or related compounds of the present invention was well mixed. After the calves finished the Supplemented EXAMPLE 11 ration, an additional three (3) lbs of grain was given to them. Novel Compounds of the Invention in Preserving and/or Augmenting Innate Immune Defenses in 0236 Weight Measurements Fish 0237 Weights were obtained on the calves at Day -3, 0246 Fish farming, a form of aquaculture, has expanded Day +4 and Day +11. Weights varied between calves, but at about eleven percent (11%) per year over the past decade. US 2005/0250772 A1 Nov. 10, 2005

Fish farming may be the fastest growing Sector of the World health consequences. AS discussed, there may be a general food economy. In 1990, thirteen (13) million metric tons of ban on fish feed antibiotics in Europe which is anticipated to fish were produced. In comparison, thirty-one (31) million take effect on or about January, 2006. Likewise, govern metric tons of fish were produced inl 998. To this end, fish ments of other countries may seriously consider Similar farming has virtually overtaken cattle ranching as a primary legislation. The concerns of antibiotics in fish food may food source worldwide. About eighty-five percent (85%) of make immune System-Stimulating feed additives highly fish farming occurs in developing countries. For example, desirous in the aquaculture industry. AS contemplated China accounted for about twenty-one (21) million tons of herein, presently preferred embodiments of the compounds the World aquacultural output in 1998, with India placing a of the present invention may exist as preparations readily distant Second, at two (2) million tons. and widely recognized as Safe for animal feed manufacture. 0247 Among industrial nations, the United States, 0252 Fish may be a highly primitive form of vertebrates, Canada, Norway and Japan are the leaders in fish produc but they have a typical vertebrate immune System that reacts tion. In North America and Europe, catfish, trout and Salmon and protects from pathogen attack. AS in other vertebrates, account for over eighty percent (80%) of finfish production. the usual humoral and cellular responses characterize the The current American annual output of 450,000 metric tons immune System of fish. Indeed, the Similarity of immune is primarily channel catfish (Ictalurus punctatus), while the Systems among vertebrates may be well evidenced by the 400,000 tons produced in Norway is primarily Atlantic extensive use of fish as experimental models for Studying the Salmon (Salmo Salar). Farmed Atlantic Salmon harvests in vertebrate immune response. AS appreciated by those skilled the United States have grown from 12,000 metric tons in in the art, there may be a certain level equivalence of 1990 to more than 50,000 metric tons in 2003. immune System physiology and function acroSS Vertebrate 0248. Due to the crowded and stressed circumstances orders. Similar biochemical and physiological responses, typical of fish farming, disease may be a frequent and a whether in mammals and birds or in fish, may be logically catastrophic limitation in Successful fish farming. Hence, anticipated through administration of preferred embodi preferred embodiments of novel compounds of the present ments of the compounds of the present invention. invention which may augment and/or enhance immune 0253) The term “generally regarded as safe” or GRAS System function in fish and/or aquatic animals may have a may be a concept used Worldwide to delineate items Significant impact in the quantitative and qualitative yield in assessed as possible ingredients in feeds and may be con the aquaculture industry. For example, a primary practice in sidered to be safe for use in humans and the food chain. The finfish acquaculture, regardless of country, has been the GRAS concept simplifies the Safety assessment proceSS by prophylactic and episodic inclusion of antibiotics in feeds. eliminating GRAS ingredients from extensive and repetitive Over thirty (30) metric tons of antibiotic active ingredient assessment. AS appreciated, Substances classified as GRAS and about 6.5 to 14.3 metric tons of tetracycline were used include the leaves of monocotyledonous plants which, as in American fish feeds during 1999 alone. Pundits of this disclosed herein, contain the compounds comprising the practice have Suggested that major negative public health present invention. and ecological impacts may stem from this extensive use of 0254. Since the novel compositions of phenolic com antibiotic agents in the aquaculture industry. pounds and precursors of phenolic compounds of the present 0249. These repercussions may include, for example and invention are configured to preserve and/or augment innate not by way of limitation, undesirable environmental conse immune defenses, it will be readily appreciated that a quences and human pathogens that may be resistant to method for preserving and/or augmenting innate immune particular antibiotics or classes of antibiotics. Indeed, defenses includes phenolic compounds belonging to related escaped fish in the Broughton Archipelago, Queen Charlotte chemical families of which 6-MBOA is a member as Strait, Canada, have been found with bacteria known to described hereinabove and in the associated Figures. It is cause Several human maladies, whereas the pathogens tested intended, therefore, that the examples provided herein be as being resistant to ten (10) different antibiotics. Viewed as exemplary of the principles of the present inven tion, and not as restrictive to a particular structure or method 0250) About thirty percent (30%) of antibiotic-laden feed for implementing those principles. used in fish farms may actually be uneaten. This antibiotic laden feed instead may move directly into the wild or natural 0255. It will be further appreciated that the novel com food chain where it has been documented to kill natural positions of phenolic compounds and precursors ofphenolic marine algae, other plants and beneficial bacteria, as well as compounds belonging to related chemical families as causing deformities in halibut larvae. Additional evidence defined herein and in the associated Figures of which that antibiotics go beyond the confines of fish farms may be 6-MBOA is a member, may be administered in any manner shown in the seventy-five percent (75%) or more of wild known to those ordinary skill in the art, including but not fishes caught nearby with antibiotics in their flesh. limited to, oral, parenteral, Sublingual, topical, transdermal, intramuscular, or inhalation, and may also contain excipients 0251 The increasing occurrence of antibiotic-resistant chosen in accordance with the dosage form adopted. More bacteria may also contribute to increased antibiotic use, and over, the dosage of the extract compositions given to an therefore greater environmental and human health risks asSociated with aquaculture. Preferably, antibiotic use in individual may vary on the basis of Several considerations aquaculture may be minimized if the immune Systems of without departing from the Spirit and Scope of the present cultured fish can be somehow enhanced to levels that invention and will, accordingly, depend on the targeted improve disease resistance. Moreover, the European Union individual's particular case to be treated. has begun to mandate lower permissible levels of antibiotics 0256 From the above discussion, the present invention in fish feed due to concerns over environmental and human provides novel compositions and methods of preserving US 2005/0250772 A1 Nov. 10, 2005 22 and/or augmenting innate immune defenses in animals and away from touch), but the additional challenge of receiving humans using phenolic compounds and precursors of phe a halter for the first time, and being led/pulled into the arena nolic compounds belonging to related chemical families of and tied up appeared to be a significant StreSSor. It was which 6-MBOA is a member. It is contemplated by the observed that the Supplement treated calves were calmer inventors that presently preferred embodiments of the during the StreSS challenge. present invention for preserving and/or augmenting immune 0261) The first two groups of calves that were handled System defense may be administered to animals (e.g., fish, were those calves that had been fed compounds of the bird, reptile and mammal) and humans. Moreover, mammals present invention (e.g., ground corn leaves), followed by the may include bear (e.g., polar, grizzly, black, brown, panda control group of calves, which had not received corn leaves, and the like), buffalo, canine (e.g., wolf, fox, dog, coyote and just an equivalent amount of grain. There were differences in the like), cow, deer (e.g., deer, moose, elk, antelope, caribou behavior during the chute manipulations between groups and the like), elephant, feline (e.g., cat, lion, tiger, cheetah, (controls were harder to get into the chute, and proceeded to cougar, leopard, jaguar, lynx and the like), giraffe, goat, buck and thrash more), and it was observed that following hippopotamus, horse, kangaroo, koala, manatee, primate being tied up to the fence, the Supplemented calves did not (e.g., monkey, gorilla, chimpanzee, gibbon, baboon, oran thrash around nor pull against the lead after a few minutes. gutan, lemur, tamarin and the like), rhinoceros, rodent (e.g., In contrast, the control group pulled back from the fence, mouse, rat, mole and the like), Seal, sheep, Sow, walrus, threw themselves against the fence and on the ground, and weasel (e.g., otter, beaver, badger, mink and the like), whale, continued this behavior for most of the stress time period. yak and Zebra. The calves whose grain was Supplemented with composi 0257). While specific dose levels were used in the tions of the present invention settled down after a few Examples and based upon absolute dosage ranges (e.g., 5 minutes, andjust stood there, watching the thrashing control mcg to 60 mg) or dosage ranges individualized to body group of calves. Moreover, the Supplemented calves were weight (e.g., 5 mg/kg to 40 mg/kg), the dosages are merely easier to untie and lead out of the arena at the end of the exemplary of Some of the presently preferred embodiments StreSS period, while the control group of calves were difficult of the present invention and are not intended to be restrictive to approach and generally resisted being untied and moved. thereof. Whereas, effective dose levels may vary to a large extent, and preferred dose levels may vary with the condi EXAMPLE 13 tions being treated and the size or Sex of the perSon being treated. To this end, dose levels do not appear to be critical Anxiety Behavior of High Spirited Dressage Horse as long as an effective amount is given. 0262. A high performance dressage horse exhibiting Sig 0258 Unlike prior art techniques or methods, the present nificant Startle and resultant aggressive behaviors was fed 40 invention provides novel chemical compositions derived, mg/kg compounds of the present invention (e.g., ground isolated and/or extracted from monocotyledonous plants or corn leaves) mixed in grain daily and observed for behav by chemical Synthesis, and methods for using Said compo ioral changes. Within 3 days of Supplementation, the horse Sitions to: (1) preserve and augment innate immune defenses was noticeably quieter and easier to handle, while maintain in humans and animals; (2) elevate levels of indoleamine, ing energy and performance ability. The owner has main for example, Serotonin and melatonin in humans and ani tained the horse on 20 mg/kg for a long period and reports mals; (3) function as Serotonin and/or melatonin analogs continued calming effects. and/or agonists in mammals; (4) Suppress the immunoregu latory effects of the hypothalamic-pituitary-adrenal (HPA) EXAMPLE 1.4 axis; (5) counteract the immunosuppressive effects of glu cocorticoid hormones (e.g., cortisol); (6) counteract the Anxiety Associated with Weaning Horses immunosuppressive effects of catecholamine compounds (e.g., epinephrine and norepinephrine); (7) enhance T-cell 0263. A Montana horse rancher participated in a study proliferation and macrophage antigen presentation and (8) wherein the rancher had a historically challenging experi decrease complications associated with immune System ence with weaning horses, with most foals becoming ill just abnormality (e.g., increased risk of infection and autoim after weaning, often with Severe consequences including mune reactions). two deaths in the 2002 weaning season. In the 2003 season, the mares and foals were fed the equivalent of 20-40 mg/kg EXAMPLE 12 compounds of the present invention (e.g., ground corn Anxiety Behavior in Calves Placed in Stressful leaves) mixed in grain before and after weaning for several Situations days. Observers noted there was leSS Vocalization at the time 0259 Compounds of the present invention which were of Separation of the mares and foals, and no illness was obtained from the leaves of a monocotyledonous plant were detected in the foals following weaning. Successfully mixed with grain and fed to calves with a weight range of 160-300 lbs. The animals were fed between EXAMPLE 1.5 20 mg/kg and 70 mg/kg of compounds of the present invention (e.g., ground corn leaves) daily and the calves, Novel Compounds of the Invention in Tests of readily consumed the rations. There was no difference in Anxiety in Mice weight gain between control and Supplement fed calves over 0264. Tests for examining anxiety in mice involve sub a one month period. jecting the animals to a forced confrontation with a novel 0260 All of the calves showed anxiety behaviors in the environment and/or exposure to predator odor. The effects of Squeeze chute (i.e., reluctance to enter the chute and pulling anti-anxiety agents on the anxiety level and emotional US 2005/0250772 A1 Nov. 10, 2005 23 reactivity of the animals may be examined by comparing a paper towel. A predator Scent was introduced into a portion behavioral responses in control and experimental groups of of the box by placing fresh dog feces, from a three year old the animals. domestic dog, in the center of the third quadrant of the box. 0265 Female albino inbred strain mice, Balb/c (Charles 0271 Mice were tested in the early dark phase between River, Inc.), at about twelve to fourteen (12-14) weeks of age about 7:00 p.m. and about 10:00) p.m. A test consisted of were Selected for use in the present exemplary Study. AS gently placing a mouse into the Safe corner of the box. This appreciated by those skilled in the art, Balb/c mice are often began a test phase where certain behavior responses of the used in anxiety testing because they are known to be mouse were recorded over a five (5) minute time period. especially Sensitive to anxious Settings and behavioral During the test phase, the mouse was allowed to move about responses may be easily observed in these animals. the box at will. 0266. It was anticipated that Balb/c mice that were sub 0272. The behavioral response measures used to assess jected to an anxiety-inducing Situation, after being fed a the reaction of the mice to the novel environment and preparation of immature corn leaves containing 6-MBOA or predator odor were the time (in Seconds) for: (1) the mouse related compounds of the present invention (i.e., 6-MBOA to attempt to explore open field, (2) the number of entries of mice), would demonstrate diminished behavioral responses the mouse into peripheral portion of the open field, (3) the asSociated with anxiety. For instance, it was expected that number of entries into the central portion of the open field, they would exhibit less timidity, be bolder in exploring their (4) the number of approaches directly towards predator odor environment, and demonstrate leSS fear of potential preda object, (5) the number of Stretching and/exploratory behav tors. In contrast, it was anticipated that Balb/c mice, not fed iors, (6) the number of exhibits of defensive tail postures, preparations immature corn leaves containing 6-MBOA or and (7) the number of “frozen” posturing sessions. related compounds of the present invention (i.e., non-6- MBOA mice) would demonstrate, to a greater degree than 0273. The study results, presented as meanistandard the 6-MBOA mice, behavioral responses associated with error of the mean, for n=7, except otherwise indicated, are anxiety. presented in Table 7, as follows: 0267 Study Design 0268. In the present study, fourteen (14) female Balb/c Group 1 Significance mice were divided into two groups of seven (7) each. These Behavioral Non- Group 2 (paired t groups comprised the following characteristics: (1) Group 1 Response 6-MBOA 6-MBOA test & included a control group of non-6-MBOA mice and (2) Measure mice mice ANOVA) Group 2 included a group of dosed 6-MBOA mice (e.g., fed Time to initial 40.67 8.93 19.33 + 9.00 p < 0.21; *NS the preparation of immature corn leaves containing attempt to explore n = 6 n = 6 6-MBOA or related compounds of the present invention). open field Number of entries The non-6-MBOA mice were not given or dosed with into peripheral 6-MBOA. All were placed, three to five (3-5) per cage, portions of open under a light/dark cycle of 1%2 hours. Lights came on at 6 field: am and went off at 6 pm. Temperature was maintained Entries into 2.83 - 0.26 3.2 + 0.26 p < 0.51; NS between about 21° C. and about 23° C. (about 70 F. and nearest peripheral about 73 F.). Commercial rodent food pellets and water section were freely available to both Group I and Group II. Entries into 2.O. O.22 2.83 + 0.26 p < 0.05; second nearest Significant 0269. For each of the five days prior to exposure to the peripheral anxiety test procedure, a preparation of immature corn section Entries into O.83 - 0.40 2.0 + 0.49 p < 0.61; NS leaves containing 6-MBOA or related compounds of the third nearest present invention at an appropriate concentration (e.g., 40 peripheral mg/kg/mouse/day) was administered orally to the mice in section Group 2 via feed. For each of the five days prior to exposure Entries into O.33 - 0.28 1.17 + 0.46 p < 0.05; furthest peripheral Significant to the anxiety test procedure, the mice in Group 1 had food section prepared the same way, without the addition of immature Number of entries 1.17 O.34 1.68 + 0.28 p < 0.21; NS corn leaves containing 6-MBOA or related compounds of into central re the present invention. gion of open field Number of ap- O.67 + O.28 1.33 + 0.28 p < 0.01; 0270. After the five days, anxiety testing was performed proaches directly Significant towards predator using an open field testing apparatus wherein the mice, object normally housed in Small defined cage environments, were Number of stretch- 3.17 O.70 8.67 + 0.94 p < 0.01; Suddenly introduced into a larger plastic box having lateral ing/exploratory Significant dimensions of between about 35 cm and about 76.2 cm and behaviors Number of de- 2.17 O.63 1.17 + 0.46 p < 0.21; NS a vertical dimension of about 20 cm. Peripheral sections fensive tail were marked on the box and measured about 18 cm each. postures Preferably, paper toweling, which covered the box floor, was Number of “frozen 95.83 + 22.58 10.83 + 9.16 p < 0.01; replaced before introduction of each test animal to remove posturing sessions Significant scent residues from the previous tests. The box was illumi nated with a 100 W light bulb except in one corner, which *Not significant was designated the “safe corner” and which was shaded by US 2005/0250772 A1 Nov. 10, 2005 24

0274 Statistical Analysis corn leaves containing 6-MBOA or related compounds of the present invention (6-MBOA mice), as compared the 0275. The significance of behavioral response differences non-6-MBOA mice. The differences in the mean values for were analyzed using paired t-tests and analysis of variance the all of the behavioral response measurements were con (ANOVA) with p-0.05. Sistent with initial expectations. In five of the ten response 0276 Results categories, the differences in mean values was Statistically Significant. Mice exposed to immature corn leaves contain 0277 Referring generally now to Table 7, as was ing 6-MBOA or related compounds of the present invention expected, the 6-MBOA mice of Group 2 exhibited less had fewer anxiety related behavioral responses to the novel timidity, were bolder in exploring the open field environ environments and fear inducing objects, Suggesting that ment, and demonstrated leSS fear of the potential predator immature corn leaves containing 6-MBOA or related com scent. The control mice of Group 1 (non-6 MBOA mice) pounds of the present invention act as anti-anxiety agents in demonstrated a much longer mean time, as noted in Table 7, Balb/c mice. to initiate an attempt to explore the open field. Though the relatively large variability renders the results not signifi EXAMPLE 16 cantly different by the measure used here, the difference in calculated means is in the anticipated direction, Suggesting Anxiety ASSociated with Transportation and Loud that the 6-MBOA mice may experience less hesitation in Noises exploring a new environment than the non-6 MBOA mice. 0285) A large, 12 year old mixed breed dog was given 0278 Table 7 also suggests that the 6-MBOA mice of compounds of the present invention (e.g., ground corn Group 2 had a greater tendency to explore the peripheral leaves) in his daily ration of food. Prior to receiving the regions of the new open field environment. Specifically, compounds of the invention the dog showed marked anxiety more mean entries were recorded for the 6-MBOA mice than in multiple situations, (i.e., running away when the vacuum the non-6 MBOA control mice into every peripheral quad was near, trembling and destructive behavior when trans rant. In the case of the Second quadrant and the most distant ported or left in a car). After receiving compounds of the quadrant, quadrant four, the differences in mean values were present invention (30-40 mg/kg) for a few days, the dog's Significant. reaction to Stressors was reduced Significantly, especially the 0279 Table 7 further suggests a slightly larger tendency agitation associated with being transported. In general, the of the 6-MBOA mice of Group 2 to explore the central dog was calmer While maintaining his normal energy level. region of the open field than the non-6-MBOA mice of Group, though the difference in mean values was not sta EXAMPLE 1.7 tistically significant. Both groups seemed to prefer the Calming Effect on Young Male Dog periphery of the open field over the central region. 0286 A young, pure-bred Labrador retriever male obses 0280 A significant difference is seen, as set forth in Table Sively collected items throughout the home of his owners, 7, in the larger mean tendency of the Group 2 mice destroying the item if the owners didn't timely intervene. (6-MBOA mice) to approach the predator scent object. This Within a few days of receiving compounds of the present implies that the 6-MBOA mice were less fearful than the invention (e.g., corn leaves) in the dogs food (30-40 control mice of Group 1. mg/kg), this behavior, along with frequent barking and tail 0281. A significant difference is also seen in the larger chasing, ceased. The observational results showed the young mean number of Stretching postures observed for the dog as being calmer. 6-MBOA mice in comparison with the non-6-MBOA mice. 0287. As will be appreciated, the present invention may AS appreciated by those skilled in the art, Stretch postures be embodied in other specific forms without departing from are usually a preliminary to locomotion. This again indicates its Spirit or essential characteristics. The described embodi less fear of the novel environment in the 6-MBOA mice over ments and examples are to be considered in all respects only the non-6-MBOA mice. as illustrative, and not restrictive. The Scope of the invention 0282 Greater values for the mean number of defensive is, therefore, indicated by the appended claims, rather than postures illustrated by the non-6-MBOA of Group 1 versus by the foregoing description. All changes which come the 6-MBOA mice of Group 2, though not statistically within the meaning and range of equivalency of the claims Significant, may again indicate leSS fear and anxiety in the are to be embraced within their Scope. 6-MBOA mice. What is claimed and desired to be secured by United States 0283 Finally, the mean tendency of the non-6-MBOA Letters Patent is: control mice of Group 1 to initiate a frozen posture, which 1. A proceSS for calming and reducing anxiety in humans is interpreted as a defensive or fearful response to novel and animals by the administration of an effective amount of stimulation, was significantly greater than for the 6-MBOA one or more chemical compositions defined as: mice of Group 2. The fewer frozen postures and greater tendency of the 6-MBOA mice to move about and explore new environments and objects is therefore related to leSS anxiety and fear. 0284 Generally, the results of this exemplary study showed a decrease in behavioral responses associated with anxiety and fear in the mice fed the preparation of immature US 2005/0250772 A1 Nov. 10, 2005 25

wherein “R” represents C-C alkoxy, with the provision 10. A process as defined in claim 9, wherein said har the R is in the 4 or 5 ring position; Vested plant is dried at a temperature in the range of between about 40° C. and about 45° C. wherein “n” represents one of the integers 0, 1 or 2, 11. A proceSS as defined in claim 9, wherein Said dried wherein “B” represents H and “A” represents -OH, harvested plant contains phenols in total amounts greater -NH, or NHCR, where R' denotes C-C, alkyl; and than 17.0 mg/gm (dry weight). “BA” represents 12. A process as defined in claim 9, wherein Said dried harvested plant contains combined amounts of 4-hydroxy cinnamic acid and 4-hydroxy-3-methoxycinnamic acid O H OH O OH totaling no more than 1.5 mg/gm (dry weight). | | || | 13. A process as defined in claim 8, wherein Said har -C-N- or ---- Vested plant is immature corn, Zea mayS. H 14. A proceSS as defined in claim 13, wherein Said immature corn has been grown to a height between about 45 centimeters and about 122 centimeters. or pharmaceutically acceptable Salts thereof: 15. A process as defined in claim 13, wherein said 2. A proceSS as defined in claim 1, further comprising the immature corn has been grown to a height that does not administration of an effective amount of one or more chemi exceed between about 30 centimeters and about 45 centi cal compositions defined as: meterS. 16. A proceSS as defined in claim 13, wherein Said immature corn has been grown for less than ten weeks after planting. 17. A proceSS as defined in claim 1, wherein Said chemical R C R2 composition is administered in a manner Selected from the group consisting of: (1) orally, in the form of tablets, N O capsules, Suspensions, Solutions and other means Suitable for R3 ingestion, including Sublingual dosage forms; (2) intranasal administration; (3) transmucosal administration; (4) parenteral injection, in the form of Subcutaneous, intramus wherein “R” is selected from the group consisting of H cular, intravenous; (5) implant for Sustained release; and (6) and OCH; transdermal patch. 18. A proceSS for calming and reducing anxiety in humans wherein “R” is selected from the group consisting of H and animals by the administration of an effective amount of and Glucose (as a glucoside) one or more chemical compositions defined as: wherein “R” is selected from the group consisting of H, OH, and OCH; or pharmaceutically acceptable Salts thereof. 3. A process as defined in claim 1, wherein Said admin istered chemical composition comprises a daily dosage of between about 5 mcg and about 60 mg. 4. A proceSS as defined in claim 1, wherein the adminis tered chemical composition comprises a daily dosage of 15 mg. wherein “R” represents C-C alkoxy, with stipulation 5. A proceSS as defined in claim 1, wherein Said admin that R is in the 4 or 5 ring position; istered chemical composition comprises a daily dosage wherein “n” represents one of the integers 0, 1 or 2, individualized to body weight of between about 5 mg/kg and about 40 mg/kg. or pharmaceutically acceptable Salts thereof. 6. A proceSS as defined in claim 1, wherein Said admin 19. A process as defined in claim 18, wherein one of said istered chemical composition comprises a daily dosage chemical compositions comprise 6-methoxy-2,3-benzox individualized to body weight of 25 mg/kg. aZolinone defined as: 7. A proceSS as defined in claim 1, wherein Said animal is Selected from the group consisting of fish, bird, reptile, and mammal. CHO 8. A process as defined in claim 1, wherein at least one of Said chemical compositions is obtained from one or more monocotyledonous plants Selected from the group consisting of corn, wheat, barley, rye, oats, rice, Sorghum, millet, bamboo, Job’s Tears, barley-like grasses, and wild grasses, by growing the plant to an immature life history Stage and or pharmaceutically acceptable Salts thereof. harvesting the plant. 20. A process as defined in claim 18, wherein one of said 9. A process as defined in claim 8, wherein said harvested chemical compositions comprise 5-methoxy-2,3-benzox plant is dried. aZolinone defined as: US 2005/0250772 A1 Nov. 10, 2005 26

intramuscular, intravenous; (5) implant for Sustained release; and (6) transdermal patch. O 36. A process for calming and reducing anxiety in humans YCE and animals by the administration of an effective amount of M one or more chemical compositions defined as: CHO H R or pharmaceutically acceptable Salts thereof. 21. A proceSS as defined in claim 20, wherein Said administered chemical composition comprises a daily doS N O age of between about 5 mcg and about 60 mg. R3 22. A process as defined in claim 19, wherein the admin istered chemical composition comprises a daily dosage of 15 mg. wherein “R” is selected from the group consisting of H 23. A proceSS as defined in claim 18, wherein Said and OCH; administered chemical composition comprises a daily doS age individualized to body weight of between about 5 mg/kg wherein “R” is selected from the group consisting of H and about 40 mg/kg. and Glucose (as a glucoside) 24. A proceSS as defined in claim 18, wherein Said wherein “R” is selected from the group consisting of H, administered chemical composition comprises a daily doS OH, and OCH; or age individualized to body weight of 25 mg/kg. 25. A proceSS as defined in claim 18, wherein Said animal pharmaceutically acceptable Salts thereof. is Selected from the group consisting of fish, bird, reptile and 37. A process as defined in claim 36, wherein said mammal. administered chemical composition comprises a daily dos 26. A process as defined in claim 18, wherein at least one age of between about 5 mcg and about 60 mg. of Said chemical compositions is obtained from one or more 38. A process as defined in claim 36, wherein said monocotyledonous plants Selected from the group consisting administered chemical composition comprises a daily dos of corn, wheat, barley, rye, oats, rice, Sorghum, millet, age individualized to body weight of between about 5 mg/kg bamboo, Job’s Tears, barley-like grasses, and wild grasses, and about 40 mg/kg. by growing the plant to an immature life history Stage and 39. A process as defined in claim 36, wherein at least one harvesting the plant. of Said chemical compositions is obtained from one or more 27. A proceSS as defined in claim 26, wherein Said monocotyledonous plants Selected from the group consisting harvested plant is dried. of corn, wheat, barley, rye, oats, rice, Sorghum, millet, 28. A process as defined in claim 27, wherein said bamboo, Job’s Tears, barley-like grasses, and wild grasses, harvested plant is dried at a temperature in the range of by growing the plant to an immature life history Stage and between about 40° C. and about 45° C. harvesting the plant. 29. A process as defined in claim 27, wherein said dried 40. A process as defined in claim 39, wherein said harvested plant contains phenols in total amounts greater harvested plant is dried. than 17.0 mg/gm (dry weight). 41. A process as defined in claim 40, wherein Said 30. A process as defined in claim 27, wherein said dried harvested plant is dried at a temperature in the range of harvested plant contains combined amounts of 4-hydroxy between about 40° C. and about 45° C. cinnamic acid and 4-hydroxy-3-methoxycinnamic acid 42. A proceSS as defined in claim 40, wherein Said dried totaling no more than 1.5 mg/gm (dry weight). harvested plant contains phenols in total amounts greater 31. A proceSS as defined in claim 26, wherein Said than 17.0 mg/gm (dry weight). harvested plant is immature corn, Zea mayS. 43. A process as defined in claim 40, wherein said dried 32. A proceSS as defined in claim 31, wherein Said harvested plant contains combined amounts of 4-hydroxy immature corn has been grown to a height between about 45 cinnamic acid and 4-hydroxy-3-methoxycinnamic acid centimeters and about 122 centimeters. totaling no more than 1.5 mg/gm (dry weight). 33. A process as defined in claim 31, wherein said 44. A process as defined in claim 39, wherein said immature corn has been grown to a height that does not harvested plant is immature corn, Zea mayS. exceed between about 30 centimeters and about 45 centi 45. A process as defined in claim 44, wherein Said meterS. immature corn has been grown to a height between about 45 34. A proceSS as defined in claim 31, wherein Said centimeters and about 122 centimeters. immature corn has been grown for less than ten weeks after 46. A process as defined in claim 44, wherein Said planting. immature corn has been grown to a height that does not 35. A process as defined in claim 18, wherein said exceed between about 30 centimeters and about 45 centi chemical composition is administered in a manner Selected meterS. from the group consisting of: (1) orally, in the form of 47. A process as defined in claim 44, wherein Said tablets, capsules, Suspensions, Solutions and other means immature corn has been grown for less than ten weeks after Suitable for ingestion, including Sublingual dosage forms, planting. (2) intranasal administration; (3) transmucosal administra 48. A process for producing one or more chemical com tion; (4) parenteral injection, in the form of Subcutaneous, positions defined as: US 2005/0250772 A1 Nov. 10, 2005 27

53. A process as defined in claim 52, wherein said

immature corn has been grown to a height between about 45 centimeters and about 122 centimeters. 54. A process as defined in claim 52, wherein said immature corn has been grown to a height that does not exceed between about 30 centimeters and about 45 centi meterS. wherein “R” represents C-C alkoxy, with the provision 55. A process as defined in claim 52, wherein said the R is in the 4 or 5 ring position; immature corn has been grown for less than ten weeks after planting. wherein “n” represents one of the integers 0, 1 or 2, 56. A proceSS as defined in claim 51, further comprising wherein “B” represents H and “A” represents -OH, the Step of drying the monocotyledonous plant. -NH, or NHCR, where R' denotes C-C, alkyl; and 57. A process as defined in claim 56, wherein said drying “BA” represents is at a temperature in the range of between about 40 C. and about 45 C. to produce a dried harvested plant. O H OH O OH 58. A process as defined in claim 57, wherein said dried | | || | harvested plant contains phenols in total amounts greater -C-N- or ---- than 17.0 mg/gm (dry weight). H 59. A process as defined in claim 57, wherein said dried harvested plant contains combined amounts of 4-hydroxy cinnamic acid and 4-hydroxy-3-methoxycinnamic acid or pharmaceutically acceptable Salts thereof, the proceSS totaling no more than 1.5 mg/gm (dry weight). comprising the Steps of: 60. A process for producing one or more chemical com identifying a Source of the chemical compositions, and positions defined as: obtaining at least one chemical compositions from the SOUICC. 49. A proceSS as defined in claim 48, further comprising the production of one or more chemical compositions defined as:

R C R2 wherein “R” represents C-C alkoxy, with stipulation that R is in the 4 or 5 ring position; N O wherein “n” represents one of the integers 0, 1 or 2, R3 or pharmaceutically acceptable Salts thereof, the proceSS comprising the Steps of: wherein "R" is selected from the group consisting of H and OCH; identifying a Source of one or more of the chemical wherein “R” is selected from the group consisting of H compositions, and and Glucose (as a glucoside) obtaining at least one of the chemical compositions wherein “R” is selected from the group consisting of H, from the Source. OH, and OCH; or 61. A process as defined in claim 60, wherein one of Said chemical compositions comprise 6-methoxy-2,3-benzox pharmaceutically acceptable Salts thereof. aZolinone defined as: 50. A process as defined in claim 48, wherein the source having one or more of the chemical compositions comprises a monocotyledonous plant Selected from the group consist CHO O ing of corn, wheat, barley, rye, oats, rice, Sorghum, millet, bamboo, Job’s Tears, barley-like grasses, and wild grasses. 51. A process as defined in claim 50, wherein the step of obtaining at least one chemical composition further com prises the Steps of growing the monocotyledonous plant; and or pharmaceutically acceptable Salts thereof harvesting the plant at an immature State. 62. A process as defined in claim 60, wherein one of Said 52. A proceSS as defined in claim 51, wherein Said chemical compositions comprise 5-methoxy-2,3-benzox monocotyledonous plant is immature corn, Zea mayS. aZolinone defined as: US 2005/0250772 A1 Nov. 10, 2005 28

pharmaceutically acceptable Salts thereof, the process comprising the Steps of: identifying a Source of one or more of the chemical compositions, and CHO obtaining at least one of the chemical compositions from the Source. or pharmaceutically acceptable Salts thereof. 74. A process as defined in claim 73, wherein the source 63. A process as defined in claim 60, wherein the source having one or more of the chemical compositions comprises having one or more of the chemical compositions comprises a monocotyledonous plant Selected from the group consist a monocotyledonous plant Selected from the group consist ing of corn, wheat, barley, rye, oats, rice, Sorghum, millet, ing of corn, wheat, barley, rye, oats, rice, Sorghum, millet, bamboo, Job’s Tears, barley-like grasses, and wild grasses. bamboo, Job’s Tears, barley-like grasses, and wild grasses. 75. A process as defined in claim 74, wherein the step of 64. A process as defined in claim 60, wherein the step of obtaining at least one chemical composition further com obtaining at least one chemical composition further com prises the Steps of prises the Steps of growing the monocotyledonous plant; and growing the monocotyledonous plant; and harvesting the plant at an immature State. harvesting the plant at an immature State. 65. A process as defined in claim 64, wherein said 76. A process as defined in claim 75, wherein said monocotyledonous plant is immature corn, Zea mayS. monocotyledonous plant is immature corn, Zea mayS. 66. A process as defined in claim 65, wherein said immature corn has been grown to a height between about 45 77. A process as defined in claim 76, wherein said centimeters and about 122 centimeters. immature corn has been grown to a height between about 45 centimeters and about 122 centimeters. 67. A process as defined in claim 65, wherein said immature corn has been grown to a height that does not 78. A process as defined in claim 76 wherein said imma exceed between about 30 centimeters and about 45 centi ture corn has been grown to a height that does not exceed meterS. between about 30 centimeters and about 45 centimeters. 68. A process as defined in claim 65, wherein said 79. A process as defined in claim 76, wherein said immature corn has been grown for less than ten weeks after immature corn has been grown for less than ten weeks after planting. planting. 69. A process as defined in claim 64, further comprising 80. A process as defined in claim 75, further comprising the Step of drying the monocotyledonous plant. the Step of drying the monocotyledonous plant. 70. A process as defined in claim 69, wherein said drying 81. A proceSS as defined in claim 80, wherein Said drying is at a temperature in the range of between about 40 C. and is at a temperature in the range of between about 40 C. and about 45 C. to produce a dried harvested plant. about 45 C. to produce a dried harvested plant. 71. A process as defined in claim 70, wherein said dried harvested plant contains phenols in total amounts greater 82. A process as defined in claim 81, wherein said dried than 17.0 mg/gm (dry weight). harvested plant contains phenols in total amounts greater 72. A process as defined in claim 70, wherein said dried than 17.0 mg/gm (dry weight). harvested plant contains combined amounts of 4-hydroxy 83. A process as defined in claim 81, wherein said dried cinnamic acid and 4-hydroxy-3-methoxycinnamic acid harvested plant contains combined amounts of 4-hydroxy totaling no more than 1.5 mg/gm (dry weight). cinnamic acid and 4-hydroxy-3-methoxycinnamic acid 73. A process for producing one or more chemical com totaling no more than 1.5 mg/gm (dry weight). positions defined as: 84. A process for producing one or more chemical com positions defined as: R

N O

3

wherein “R” is selected from the group consisting of H wherein “R” represents C-C alkoxy, with the provision and OCH; the R is in the 4 or 5 ring position; wherein “R” is selected from the group consisting of H wherein “n” represents one of the integers 0, 1 or 2, and Glucose (as a glucoside) wherein “B” represents H and “A” represents -OH, wherein “R” is selected from the group consisting of H, -NH, or NHCR, where R' denotes C-C, alkyl; and OH, and OCH; or “BA” represents US 2005/0250772 A1 Nov. 10, 2005 29

identifying a monocotyledonous plant Source of one or more of the chemical compositions, O H OH O OH | | || | growing the monocotyledonous plant for a period less - C-N- or -C-C-N- than ten weeks after planting and to a height between about 45 centimeters and about 122 centimeters, harvesting the monocotyledonous plant, drying the monocotyledonous plant at a temperature in the range of between about 40 C. and about 45 C. to or pharmaceutically acceptable Salts thereof, the proceSS produce a dried harvested plant. comprising the Steps of: k k k k k