HK1153395A - Fertilized egg isolate and use thereof - Google Patents

Description

fertilized egg isolate and use thereof

Cross Reference to Related Applications

This application claims priority to U.S. provisional application No.61/020,541 filed on 11/2008 and U.S. provisional application No.61/033,184 filed on 3/2008, which are incorporated herein by reference.

Technical Field

The present invention relates to fertilized egg isolates, including fertilized egg isolates prepared by any of the methods set forth in this specification, and their use for the treatment of mental health disorders.

Background

Major depression (also known as major depression, clinical depression, unipolar depression, and unipolar disorder) is common in the general population. Recent data in North America has shown a lifetime risk of major depression in adults of 14.5% and an annual incidence of 8.1% (results from the 2004National surface on Drug Use and Health: National definitions; reviews of 9/8/2005; Department of Health and Human Services. Substance Abstract of Mental Services Administration of Applied students).

The mean duration of a depressive episode with modern therapy is about 16 weeks, but some data show a longer duration of about 6-8 months, but this is much shorter than about 18 months prior to antidepressant treatment (Kendler, McLeod, Patten).

Antidepressants have a very positive impact on the treatment of depression and on the reduction of patient suffering. Depressed patients often have impaired function and often have other unhealthy conditions, such as substance abuse, actually due to depression. Depression can cause an increased burden on health authorities and can have devastating effects on community structure and socioeconomic performance.

The cause of depression is not fully understood. Dysregulation of monoamine synthesis and activity has been the leading etiological theory of depression for the past decades, and the support for this notion is reinforced by the effectiveness of drug therapies that enhance monoamine activity, particularly those of 5-hydroxytryptamine-and/or norepinephrine. However, any given antidepressant is effective in only a small group of depressed patients, and often only partially. Current therapies performed in investigational controlled trials using selected samples are effective in only about 60% of patients, and only about half of them have complete disappearance of symptoms. This is important because the presence of residual symptoms is a strong predictor of recurrence. There are other physiological changes associated with depression that may indicate more complex interactions of etiological factors (including the function of second messengers to mediate membrane binding and intracellular processing). This led to the study of the following events: hormone pathways, such as: hypothalamic-pituitary-adrenal (HPA) axis (its activity is elevated in 20-40% of depressed patients living in the community), thyroid axis (thyroid dysfunction has not been previously detected in 5-10% of patients assessed to have depression), growth hormone, prolactin; the inflammatory process and the function of its markers (e.g., interleukin 1, interleukin 6, and tumor necrosis factor).

Most people with major Depression experience some degree of recurrence of symptoms and 20-30% present as a slow process (the level of syndrome defined as the severity of depressive symptoms is not less than 2 years (edition)).

All depressed people require continuous medication to prevent relapse and to rehabilitate them. Most depressed patients need to remain on medication to prevent relapse and further consolidate psychosocial rehabilitation. However, while a major factor in effective antidepressant treatment is maintaining an adequate dose of drug to the patient for a sufficient duration, this is often difficult. Many patients are afraid of taking existing antidepressants due to real or fantastic physical effects. Some patients prefer to use so-called natural pro-health substances and non-pharmacological interventions. Patients who are ready to take antidepressants are often faced with a number of side effects which can cause them to become non-compliant or to reject treatment altogether. For example, selective 5-hydroxytryptamine reuptake inhibitors (SSRIs) often cause many side effects such as gastrointestinal upset, headache, sleep disturbances, significant sexual dysfunction, and the like. Most antidepressants have at least some significant side effects that limit the ability of the clinician to effectively treat many patients.

Depression may be associated with other conditions and/or syndromes, including anxiety such as generalized anxiety disorder, sexual dysfunction, seasonal affective disorder, social anxiety disorder (otherwise known as social phobia), bipolar disorder, and dementia.

It is generally accepted that limited efficacy, generally unacceptable side effects and physiological factors that can induce or influence the course of depression make it necessary to continue to search for new compounds with novel pharmacological effects to address depression, a major public health problem.

Drawings

Those skilled in the art will appreciate that the following figures are for illustration purposes only. The drawings are not intended to limit the scope of the present invention in any way.

FIG. 1 is a photograph of an embryo removed from an egg according to an embodiment of the present invention.

FIG. 2 is a photograph of a freeze-dried embryo according to an embodiment of the present invention.

FIG. 3 is a photograph of a freeze-dried embryo according to an embodiment of the present invention.

FIG. 4 is a photograph of a crushed freeze-dried embryo according to an embodiment of the present invention.

FIG. 5 is a photograph of a slurry of a number of embryos according to an embodiment of the present invention.

Fig. 6 shows an HPLC chromatogram of fertilized egg isolate according to an embodiment of the present invention.

Fig. 7 shows the results of an analysis of fertilized egg isolates according to an embodiment of the present invention.

FIG. 8 is a graph showing the effect of various concentrations of fertilized egg isolate sample #20 supernatant isolate (μ g/mL) on the binding (measured as a percentage of specific binding) of radiolabeled CGP 39653 to the agonist site (ionotropic) of the NMDA glutamate receptor, and the IC of NMDA and sample #20 supernatant isolate₅₀And K_i。

FIG. 9 shows a graph of the effect of various concentrations of fertilized egg isolate sample #20 supernatant isolate (μ g/mL) on binding (measured as percent of specific binding) of radiolabeled kainic acid to the kainic acid site (ionotropic) of the kainic acid glutamate receptor, and the IC of kainic acid and sample #20 supernatant isolate₅₀And K_i。

FIG. 10 shows graphs depicting the effect of various concentrations of superisolates (μ g/mL) from fertilized egg isolate #20 on the binding (measured as a percentage of specific binding) of radiolabeled α -amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid (AMPA) to the AMPA site (ionotropic) of the AMPA receptor, and the IC of (+/-) AMPA HBr and superisolates from sample #20₅₀And Ki.

FIG. 11 shows a graph of the effect of various concentrations of superisolates (μ g/mL) from fertilized egg isolate #20 on the binding (measured as percent of specific binding) of radiolabeled MDL-105, 519 to strychnine insensitive glycine sites (ionotropic) of the NMDA glutamate receptor, and the IC of MBL-105, 519 and sample #20 superisolates₅₀And K_i。

FIG. 12 shows a graph of the effect of various concentrations of superisolates (μ g/mL) from fertilized egg isolate #20 on the binding (measured as a percentage of specific binding) of neurokinin A (NKA) to the NK2 receptor, and the IC of neurokinin A and superisolates from sample #20₅₀And K_i。

Detailed Description

According to the present invention, fertilized egg isolates and their use for the treatment of mental health disorders are described.

Fertilized egg isolate-preparation

According to one aspect of the present invention, a method is provided for preparing a chalked sample of at least one embryo or preparing a chalked sample from a portion or all of the contents of a fertilized egg. At least one fertilized egg is incubated for about 3 to about 15 days, more preferably about 3 to about 5 days, or more preferably about 6 to about 12 days, and still more preferably about 7 to about 9 days from the date of fertilization of the egg. Generally, the fertilized egg is incubated for a period of time that allows angiogenesis to begin and/or maturation of the embryo to the naked eye. The eggs may be from a variety of sources, such as birds, reptiles, or egg-producing mammals. In general, any egg from which an embryo or blood vessel to which an embryo is attached can be removed is suitable. The eggs are preferably avian eggs, and may be obtained from any avian species that has been bred for egg production, such as chickens, geese, ducks, and the like. Chicken eggs are preferred for reasons including their feasibility and ability to be produced on a large scale. Incubation can be carried out in any environment, as long as the eggs are maintained for an extended period of time at a temperature at which the embryos mature. Suitable incubation temperatures range from about 20 to about 60 deg.C, more preferably from about 25 to about 55 deg.C, and more preferably from about 35 to about 45 deg.C. Once the eggs have been incubated for a period of time, they may optionally be treated to reduce the external microbiota or to sterilize them by any suitable means, such as: washing the egg shell with a solvent such as ethanol (e.g., about 50% to about 95% ethanol solution), and then evaporating or drying the solvent; or rotating the egg under an Ultraviolet (UV) light source for a suitable period of time. Preferably, all solvent is evaporated before further processing of the eggs. The eggs are then broken to gain access to the contents. The eggs may be broken under aseptic conditions manually or using suitable mechanical equipment. This process and/or all or most of the processes described above and below may be carried out in a cooled atmosphere, such as an atmosphere at about 5 ℃.

According to one aspect of the invention, the contents of the eggs are collected in a container, such as a stainless steel container, which is preferably sterilized and/or frozen. The contents taken from the container or from the egg may optionally be subjected to a filtration treatment, for example by placing on a sieve. The mesh size is about 0.5 to about 4mm, more preferably about 1 mm. The screen is preferably sterile.

Optionally, the contents of the egg and/or some or all of the ruptured egg shell may be placed directly on the screen. Allowing the contents of the egg and/or some or all of the broken egg shells to filter on the screen for a period of time until substantially no more droplets pass through the screen. The broken egg shells may be removed from the contents of the egg before, during or after the filtration process. After filtration, the solid or solid and semi-solid retentate may comprise the embryo, vascular connective tissue, most or all of the egg white, most or all of the zona ovalis, and clear sac (clear sac). The semi-solid retentate may comprise solid material and viscous material, for example, a gelatinous material such as egg white. The retentate or semi-solid retentate optionally can be washed at least once with a suitable solvent, such as a buffer solution, sterile deionized water, or any suitable saline solution. For example, sterile Phosphate Buffered Saline (PBS) may be used.

The retentate may be collected from one egg and then freeze-dried according to the methods described herein, or the retentate may be collected from one or more eggs together and then freeze-dried according to the methods described herein.

According to another aspect of the invention, the white protein fraction and/or embryo may be substantially separated from other egg contents. The white albumen portion may be substantially separated from the other contents by any suitable method (e.g., pouring out the white albumen portion) or by suction. The embryo may be substantially separated from the white protein fraction, either manually or by other suitable methods as determined by the skilled artisan. FIG. 1 shows an example of an embryo removed from an egg and then washed with buffer. One skilled in the art will recognize that embryos can be substantially separated from the white protein fraction and other contents at the same time. For example, the embryo may be manually removed from the white albumen portion and other contents using forceps or other suitable tool. In some cases, the embryo may be manually stripped from the yolk sac, which forms part of the other contents.

After the embryo is substantially separated from the white protein portion and other contents of the egg, the embryo is optionally washed at least once with a suitable solvent (e.g., a buffer solution, sterile deionized water, or any suitable saline solution). For example, sterile Phosphate Buffered Saline (PBS) may be used.

It will be appreciated that if the contents of the egg are subjected to a filtration process, reference to the contents of the egg will actually refer to the retentate for the following method. It is also understood that whole fertilized eggs may be broken, the egg shells removed, and the whole de-shelled eggs frozen and lyophilized according to any of the methods described above and below to produce fertilized egg isolates of the present invention. Alternatively, a plurality of intact zygotes may be broken, the shells removed, all of the shelled zygotes combined and mixed into a slurry, then frozen and lyophilized according to any of the methods described above and below.

The contents of the egg or embryo are placed in at least one container that can be frozen. The container may be, for example, a test tube, a petri dish, a beaker, a stainless steel tray or a plastic container. Preferably, the contents or embryos are frozen soon after removal from the shell, e.g., within about 2 hours, more preferably within about 1 hour, even more preferably within about 0.5 hour or as quickly as possible. Depending on the length of time the contents or embryos are to be frozen, the freezing temperature may range from about-50 ℃ to about 10 ℃, more preferably from about-40 ℃ to about 5 ℃, and even more preferably from about-35 ℃ to about-25 ℃. Preferably, the contents or embryos are frozen for at least about 6 hours, more preferably at least about 12 hours, even more preferably at least about 24 hours. The frozen contents or embryos may be freeze-dried or lyophilized after a period of time. The contents or embryos may be completely frozen prior to the freeze-drying/lyophilization step. FIGS. 2 and 3 show examples of embryos after freeze-drying.

Optionally, the frozen or unfrozen contents or embryos can be combined in a suitable container (e.g., a beaker or plastic container) and mixed or blended with a suitable solvent as desired to form a slurry. The solvent may have suitable moisture to wet the mixed contents or embryos and can be frozen in a standard laboratory freezer. Suitable solvents include water, aqueous buffers, and the like. To form a slurry, the contents and/or embryos are preferably mixed. The contents or embryos may be mixed or homogenized, for example, with a hand-held mixer or other suitable tool. The slurry was then frozen and freeze dried as described above. Preferably, lyophilization is carried out at an ultimate temperature in the range of from about-80 ℃ to about-10 ℃, more preferably from about-65 ℃ to about-15 ℃, even more preferably from about-40 ℃ to about-20 ℃, at a pressure of about 500 millitorr or other suitable pressure as may be determined by the skilled artisan. Preferably, the freeze-drying process is maintained at the limiting temperature for a period of time of from about 1 to about 6 hours, more preferably from about 2 to about 5 hours, and even more preferably from about 3 to about 4 hours. The time for the entire lyophilization process is generally from about 15 to about 45 hours, more typically from about 25 to about 35 hours, and even more typically from about 28 to about 32 hours.

According to another aspect of the invention, the freeze-dried contents, freeze-dried embryos or freeze-dried slurry may be dispersed and/or comminuted as desired to form a substantially homogeneous powder. The individual or small groups of freeze-dried contents may be combined together before or after the comminuting step to form a substantially homogeneous powder. The grinding can be done mechanically, for example using a suitable machine, such as a coffee bean grinder or hammer mill, or manually using a suitable tool, such as a glass rod. FIG. 4 shows an example of a crushed freeze-dried embryo. The powder is optionally sterilized prior to storage and/or use. Suitable sterilization procedures should not adversely affect certain freeze-dried components.

According to any of the methods described herein, a preservative to control microbial growth may be mixed into the powder or concentrate prior to storage. Preservatives may also be added at another stage of manufacture, including prior to the freeze-drying or concentration steps, rather than or in addition to the powder or concentrate. Suitable preservatives include common food preservatives such as 0.5% w/w sodium benzoate and 0.2% w/w potassium sorbate. Other suitable preservatives may be selected by the skilled person.

The powder produced by the methods disclosed herein can be stored in a suitable, substantially airtight container. Suitable containers include plastic bags, pails, plastic containers, bottles, combinations thereof, and the like. For example, the powder may be packaged under controlled aseptic conditions into sterile polyethylene/polypropylene bottles with tamper-proof safety seals. The powder may be stored in a substantially dry inert gas such as nitrogen. The powder is preferably stored at room temperature or lower, for example from about 10 ℃ to about 25 ℃, more preferably from about 15 ℃ to about-20 ℃. For long term storage, the powder is preferably stored at a temperature of about-10 ℃ or less, or more preferably, about-20 ℃ or less. The powder may be stored in a substantially dry atmosphere for a period of time. The powder may also be vacuum packed.

According to another embodiment of the invention, the slurry is prepared as described above or can be prepared by: the contents or embryo of at least one fertilized egg is separated from the egg shell and the separated contents or embryo are combined in a suitable container. The separated contents or embryos may be cooled during this step. For example, the container may be placed on ice to facilitate cooling. The contents or embryos can be mixed by the methods described above to produce a slurry. FIG. 5 shows an example of a container of embryo slurry. The slurry may be freeze-dried as described above or used partially or wholly for the extraction step as described below.

The aqueous phase extraction process is carried out by mixing the slurry with an aqueous solution for a period of time. The aqueous solution may comprise water, an aqueous buffer solution or any other aqueous solvent. If the aqueous solution comprises water, the water is preferably distilled water, more preferably also deionized prior to use. For example, the water may be treated using reverse osmosis (r.o.). The slurry and aqueous solution may be mixed, for example, by stirring for a period of time ranging from about 5 to about 60 minutes, more preferably ranging from about 10 to about 45 minutes, and even more preferably ranging from about 15 to about 40 minutes. Desirably, the aqueous solution is in sufficient contact with the slurry of contents such that any substantially hydrophilic molecules in the solution are dissolved in the aqueous solution. The volume of the aqueous solution used may be substantially equal to the volume of the slurry, but may also be 1.5, 2, or even 3 times the volume of the slurry. Optionally, the mixture may be heated slightly during the mixing step. After mixing, any solid portion of the mixture may be substantially removed by any suitable means, such as centrifugation or filtration, thereby rendering the aqueous solution substantially clear. The clarified aqueous phase portion may then be frozen and freeze-dried according to the methods described herein to produce a powder, which is optionally sterilized.

According to another aspect of the invention, the slurry produced by any of the above methods may be mixed with a substantially hydrophobic solvent. The substantially hydrophobic solvent is preferably chilled. Suitable hydrophobic solvents include, for example, ether, chloroform, hexane, petroleum ether, or acetonitrile. For example, ethers, especially diethyl ether, may be used. The slurry is mixed with the hydrophobic solvent for a period of time, as described above. One skilled in the relevant art will recognize that any step of the process using a substantially hydrophobic solvent should be performed in a fume hood or similar device, and that the solvent should be kept away from an open flame or heat source. After a period of mixing, the solid portion of the mixture can be substantially removed from the solvent portion by any suitable method, such as centrifugation or filtration. The solvent fraction will comprise predominantly a hydrophobic solvent fraction and possibly also an aqueous fraction. The solvent portion can be transferred to a separatory funnel or substantially equivalent apparatus to separate the aqueous portion from the hydrophobic solvent portion. If the top layer is part of the hydrophobic solvent, it can be aspirated from the top, or removed from the separatory funnel after the bottom aqueous layer is removed. Alternatively, the bottom aqueous portion can be frozen so that the top ether layer can be poured off. The aqueous portion may be extracted multiple times, e.g., about 3 times, with the hydrophobic solvent. The volume of the hydrophobic solvent and the aqueous portion may be substantially equal, or may be 1.5 times, 2 times, or even 3 times the volume of the aqueous solvent. Other ratios may also be suitable.

After the extraction process, all hydrophobic isolates can be combined and concentrated by suitable methods. The concentrated isolate may be stored at a temperature below room temperature (e.g., about 5 ℃) in a suitable substantially air-tight container, such as a sealed vial.

According to another aspect of the invention, the slurry produced by any of the methods described above may be clarified prior to the extraction step. Preferred clarification steps include filtration methods using filters such as a sieve or filter paper or filter mat. Other clarification steps may include centrifugation. Filter aids such as Superflow DE may be added to the filtrate from the filtration step prior to further clarification^TM. Part of the resulting filtrate may be frozen in a suitable container for freeze-drying. In addition, part of the resulting filtrate may be mixed with the above-mentioned hydrophobic solvent to form an aqueous layer and a hydrophobic layer. The layers can be separated, concentrated, and stored as described herein.

Fertilized egg isolates prepared by the various methods described herein can be prepared by repeated and/or combined treatments to increase potency. For example, aqueous and/or hydrophobic solvent extractions can be repeated on the same sample to concentrate the active compound.

Fertilized egg isolate-use

Fertilized egg isolates prepared by the methods described herein or by analogous methods as would be apparent to one of skill in the art based on the teachings of the present invention may be used to treat patients suffering from mental health disorders including depressive affective disorders such as major depression, dysthymic disorder (depressive phase), bipolar disorder, depression caused by general medical conditions such as depression associated with dementia or schizoaffective disorder, substance-induced depression and seasonal affective disorder, anxiety disorders such as generalized anxiety disorder, social phobia and panic disorder, and sexual dysfunction. As will be appreciated by those skilled in the relevant art, treating such conditions as depression or anxiety may be a useful method of treating other disorders and/or symptoms associated with them (e.g., those listed above). In one embodiment, the patient is a human.

As detailed herein, it has been determined that fertilized egg isolates of the present invention can antagonize the binding interaction of certain ligands with their receptors. In particular, fertilized egg isolates of the present invention have been found to have the ability to displace glutamate from 4 major glutamate receptors. In addition, the fertilized egg isolate has the ability to displace NKA from neurokinin 2(NK2 receptor), a receptor for the neurotransmitter neurokinin a (NKA). To the best of the inventors' knowledge, this is the first time that these two groups of receptors are found to be antagonized by a single agent.

Glutamate (glutamate) is one of the most excitable substances in the human brain. Many diseases and conditions are known to be mediated by or associated with activation of one or more glutamate receptors. Such diseases and conditions include depression (see, e.g., Paul, Toro, Mathew 2005, Krystal, Sanacora 2003, Svenningsson, McNaIIy), major depression (see, e.g., Maeng, Chourbaji, Mathew 2008), anxiety (see, e.g., Rorick-Kehn), alzheimer's Disease (see, e.g., Walton, Koch, Hynd), epilepsy (see, e.g., Kew, Vincent), schizophrenia (see, e.g., McCullumsmith, Lewis, MacDonald, Javitt), brain cell loss following stroke/ischemia (see, e.g., Ginsberg, Kew), amyotrophic lateral sclerosis (Lou Gehrig's Disease) (see, e.g., mathehew 2008, Miller), manic bean poisoning (see, e.g., Spencer, ravindranaheh), autism (see, e.g., boyehr's Disease), bipolar disorder (see, e.g., mania, see, e, e.g., mania, see, bipolar disorder (see, e, e.g., cheyne), krystal). Furthermore, antagonists of glutamate receptors such as ketamine and riluzole (riluzole) have been shown to be useful in the treatment of depression (Mathew 2008), major depression (Maeng, Mathew 2008, Zarate 2004, Sanacora 2007), amyotrophic lateral sclerosis (Mathew 2008, Miller), and bipolar depression (Mathew 2008, Zarate 2005). By inhibiting the activation of one or more glutamate receptors by inhibiting glutamate binding to its receptor, conditions or diseases mediated by or associated with one or more glutamate receptors may be reduced or eliminated. As described herein, because it has been determined that fertilized egg isolates of the invention bind to one or more glutamate receptors, fertilized egg isolates of the invention are useful for treating conditions or diseases associated with or mediated by one or more glutamate receptors.

Accordingly, another aspect of the invention features a method of using fertilized egg isolate to treat a disease or disorder associated with or mediated by glutamate receptors, the method comprising the steps of: administering a therapeutically effective amount of the fertilized egg isolate to a patient in need thereof. Such glutamate receptors include ionotropic glutamate receptors such as the alpha-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid (AMPA) receptor, the kainatoecteptor and the N-methyl-D-aspartate (NMDA) receptor. Diseases or conditions associated with or mediated by glutamate receptors may include depression, major depression, anxiety, alzheimer's disease, epilepsy, schizophrenia, impairment of brain cell function following stroke/ischemia, amyotrophic lateral sclerosis (gregaria), lathyrism, autism, mental retardation, cognitive disorders, bipolar depression or mania.

A number of diseases and disorders are also known to be mediated by the NK2 receptor or associated with NK2 receptor activation. Such diseases or conditions include depression (see, e.g., Dableh, Ahlstedt, Michale, Louis, Steinberg, Salom é, Holmes, Steinberg, Husum), anxiety (see, e.g., Ahlstedt, Michale, Louis, Greibel, Steinberg, Stratton, Teixeira, Walsh, Salome, Holmes), irritable and inflammatory bowel syndromes (see, e.g., Ahlstedt, Lecci, Evangelista, Toulose), inflammatory airway diseases (see, e.g., Bai), and urinary incontinence (see, e.g., Ahlstedt, Rizzo). Furthermore, antagonists of the NK2 receptor, such as saredutant (SR 48964), have been shown to be useful in promoting antidepressant-like effects (Salom, dabley, Steinberg, Michale, Louis) and anxiolytic effects (Teixeira, Salom, Griebel, Michale, Louis) in animal models, and studies in humans have been performed. By inhibiting the activation of the NK2 receptor (e.g., inhibiting NKA binding to its receptor) by inhibiting one or more endogenous ligands of NK2, a disorder or disease mediated by the NK2 receptor or associated with the NK2 receptor can be reduced or eliminated. As described herein, since it has been determined that the fertilized egg isolate of the present invention can bind to the NK2 receptor, the fertilized egg isolate of the present invention can be used to treat a disease or disorder associated with the NK2 receptor or mediated by the NK2 receptor.

Accordingly, another aspect of the invention features a method of using fertilized egg isolate to treat a disease or disorder associated with the NK2 receptor or mediated by the NK2 receptor, the method comprising the steps of: administering a therapeutically effective amount of the fertilized egg isolate to a patient in need thereof. Diseases or disorders associated with the NK2 receptor or mediated by the NK2 receptor may include depression, anxiety, irritable bowel syndrome or urinary incontinence.

Fertilized egg isolates for use in the treatment of a disease or disorder associated with or mediated by one or more glutamate receptors and the NK2 receptor may include freeze-dried embryos, ovalbumin, and clear sacs from fertilized eggs; or it may be produced in such a manner that it does not contain a large amount of yolk from the fertilized egg. Fertilized egg isolates can be prepared according to the methods described herein or by analogous methods known to those skilled in the art. In a method for treating a disease or condition associated with or mediated by a glutamate receptor or NK2 receptor, the patient may or may not be psychologically treated concurrently with the treatment.

Another aspect of the invention is a method of treating mental health disorders comprising administering a fertilized egg isolate to a patient in need thereof, and the use of the fertilized egg isolate for such purposes.

The fertilized egg isolate described herein may be formulated and administered in a variety of dosage forms, such as those suitable for administration by the following routes: oral routes (including buccal and sublingual administration), rectal routes, intranasal routes, topical routes (including buccal, sublingual and transdermal administration), vaginal routes, rectal routes or parenteral routes (including subcutaneous, intramuscular, intravenous or intradermal routes). Particularly preferred are dosage forms suitable for administration by the oral route. Other preferred dosage forms include those suitable for administration by the vaginal or rectal route, such as suppositories.

Formulations of fertilized egg isolate suitable for oral administration may be presented as discrete units, such as capsules, tablets, microgranules, powders, granules, solutions or suspensions in aqueous or non-aqueous liquids, edible foams or whipped foods (whips), or oil-in-water or water-in-oil emulsions. Fertilized egg isolate may be combined with a suitable, orally non-toxic pharmaceutically acceptable inert carrier. For capsules, suitable capsule bodies (e.g., gelatin sheaths) may be used to encapsulate the fertilized egg isolate, either alone or in combination with one or more suitable, non-toxic, pharmaceutically acceptable inert carriers. For example, for tablets and capsules, suitable carriers can include, but are not limited to, edible carbohydrates such as starch or mannitol, flavoring agents, preservatives, dispersing agents, binding agents, and coloring agents, and can include fumed silica. The formulation may also be prepared to prolong or maintain the release of fertilized egg isolate according to methods known to those skilled in the art of pharmaceutical formulation.

The effective dose for treatment will depend on the patient. An effective dose may be about 200 to about 6000 mg/day, about 500 to about 4000 mg/day, about 750 to about 3500 mg/day, about 800 to about 3000 mg/day, about 1000 to about 2500 mg/day. For example, an effective dose may be about 2000 mg/day.

The term "treating" refers to ameliorating a disease or condition in a patient who is taking a fertilized egg isolate of the invention. The term includes ameliorating the disease or disorder, and such amelioration can be determined using standard assays known in the art. The term also includes preventing the occurrence or recurrence of the disease or disorder, e.g., in prophylactic or maintenance therapy.

Another aspect of the invention features a method for inhibiting the activity of a glutamate receptor comprising contacting the glutamate receptor with an effective amount of fertilized egg isolate. The method may be performed using ionotropic glutamate receptors, such as AMPA receptors, kainate receptors or NMDA receptors.

In addition, the invention features a method for inhibiting NK2 receptor activity, comprising contacting the NK2 receptor with an effective amount of fertilized egg isolate.

The activity of glutamate receptors or NK2 receptors can be inhibited, for example, by: inhibiting the binding of said receptor to one or more of its endogenous ligands (e.g. glutamate for the glutamate receptor; NKA for the NK2 receptor) or commercially available exogenous ligands (e.g. AMPA, NMDA, kainic acid, CGP 39653 or MDL-105, 509 for the glutamate receptor; and saredutant for the NK2 receptor). Methods for inhibiting such binding interactions and for detecting such binding inhibition are known to those skilled in the art and are also described herein. The activity of the glutamate receptor or NK2 receptor can be 100% inhibited or less than 100% (e.g., 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, or 10%) inhibited. Methods for inhibiting glutamate receptor or NK2 receptor activity can be performed in vitro (e.g., in a cell, cell lysate, or a sample containing a portion of a cell (e.g., only the relevant receptor)) or in vivo (e.g., in a patient).

According to yet another embodiment of the invention, the fertilized egg isolate may be formulated with other compounds useful for the treatment of mental health disorders. For example, the fertilized egg isolate may be formulated with a compound that inhibits the disintegration of 5-hydroxytryptamine (e.g., a monoamine oxidase inhibitor).

Other objects and advantages of the present invention will become readily apparent to those skilled in this art from the following detailed description, wherein it is shown and described only the preferred embodiments, simply by way of illustration. As will be realized, the invention is capable of other and different embodiments and its several details are capable of modifications in various obvious respects, all without departing from the invention. Accordingly, the following description and examples are to be regarded as illustrative in nature and not as restrictive.

Examples

Example 1

Fertilized chicken eggs of 8-9 days old were selected. The whole egg was sterilized with 70% ethanol and placed in a fume hood to evaporate the 70% ethanol. The eggs were broken and the contents were dropped through a sterile 1.0mm sieve. The egg shell and filtrate were discarded. The retentate, including solids and liquids, was collected and frozen on ice. The retentate was homogenized at 5 ℃. The homogenate (slurry) was poured into sterile stainless steel trays, frozen and freeze-dried. The dried product was ground to a powder using a grinder. Preservatives 0.5% w/w sodium benzoate and 0.2% w/w potassium sorbate are added to the powder and the mixture is mixed. The final powder was stored at 2-8 ℃ (short term) or-20 ℃ (long term).

Example 2

8 day old chick embryos were collected from fertilized chick eggs according to the following procedure:

(i) collecting 24 brown eggs (brown egg);

(ii) when broken, only 22 of the 24 eggs were found to be fertilized and contained developing embryos (91.6% fertilization rate);

(iii) embryos were removed and briefly washed in sterile PBS (twice) and then immediately frozen in 50mL (mL) tubes at-20 ℃ each;

(iv) then freeze-drying 10 test tubes in a freeze-dryer (shelf temperature-40 ℃, condenser temperature-52 ℃);

(v) at the end of this procedure (cycle), each embryo (still in the tube) was crushed with a sterile glass rod and the resulting powder (1.7g (g)) was packaged;

(vi) the 12 remaining embryos were collected in a small beaker and mixed with a small amount of water with a hand-held mixer;

(vii) the resulting slurry was rapidly frozen and then freeze-dried as described above.

Results

After drying, the powder (1.7g) was uniformly dispersed using a glass rod, batch A. Each embryo produced 0.17g of powder.

The dried slurry (1.6g) was uniformly dispersed using a glass rod, run No. B. Each embryo produced 0.14g of powder (humidity unknown).

Example 3

60 fertilized eggs (8-day-old embryos) were collected. The preparation of the water and ether isolates is described below.

All eggs were sprayed with 70% (v/v) ethanol to sterilize the outer surface, and then the eggs were placed in a laminar flow hood to evaporate the ethanol. The egg is broken and the embryo and associated clear sac and its clear water-like contents are carefully separated from the rest of the yolk and vascular material. All material, i.e. embryos and clear sacs, were then collected in a beaker pre-cooled on ice. After collection, the materials were mixed by means of a hand mixer to form a slurry.

The resulting slurry (about 200mL) was divided into two equal portions; one part was used for extraction with water and the other part was used for extraction with ether.

Water separation

To 100mL of the slurry was added 100mL of reverse osmosis water and the mass was stirred at room temperature for 30 minutes. The material was clarified by centrifugation and the aqueous upper layer (200mL 1.1% solids) was collected. The aqueous upper layer was then frozen and subsequently freeze-dried. The dry powder weighed 1.0 g.

Ether isolate

100mL of pre-cooled ether was added to 100mL of the slurry. The mixture was shaken at room temperature and then centrifuged. At the end of the centrifugation process (15 min, 5 ℃), the upper water/ether layer was transferred to a separatory funnel and the upper ether layer was collected by siphoning or by freezing the upper ether layer after a period of freezing at-20 ℃ which would freeze the lower cloudy water layer but not the upper yellow clear ether layer.

The lower aqueous layer was extracted again with an equal volume of diethyl ether as described above and the process was repeated three times. The ether isolate was then purified at Rotavap^TMMedium concentrated and dried to a small volume (about 1 mL). The sample was placed at 5 ℃.

Example 4

420 fertilized eggs (8-9 day old embryos) were sprayed with 70% (v/v) ethanol to sterilize the outer surface as much as possible, and then the eggs were placed in a fume hood to evaporate the ethanol.

The eggs were broken (62 of 420 found unfertilized, fertilization rate 85.2%), the embryos and associated clear sacs and their clear water-like contents were carefully separated from the rest of the yolk and vascular material, and all material, i.e. embryos and clear sacs, were collected in a beaker pre-cooled on ice. The collected material (ca. 2L) was placed at 5 ℃.

Water separation

After collection, mix by handThe combiner mixes the substances into a slurry. The resulting slurry (about 2L) was passed through a metal screen (20 mesh). The filtrate was further clarified with a paper filter pad and DE 6000. The filter cake remaining on the filter pad was discarded. Superflow DE was added to the filtrate^TMAnd then further clarified by passing the material through a filter pad. All filtration was done on a laboratory scale using a Buchner funnel (Buchner tunnel). The majority of the resulting filtrate (about 1200 and 1300mL) was separated and immediately frozen in trays for freeze drying (1.9% solids). The remaining 500mL was mixed with an equal volume of pre-cooled ether in a 1L vial and placed at 5 ℃ until two layers separated.

Ether isolate

500mL of precooled ether was added to 500mL of the slurry, and the mixture was shaken at room temperature and then centrifuged. At the end of the centrifugation process (15 min, 5 ℃), the upper water/ether layer was transferred to a separatory funnel and the upper ether layer was collected by siphoning or by freezing the upper ether layer after a period of freezing at-20 ℃ which would freeze the lower cloudy water layer but not the upper yellow clear ether layer.

The lower aqueous layer was extracted again with 300mL of diethyl ether as described above and the isolate was collected in a precooled round bottom flask. The ether isolate was then purified at Rotavap^TMMedium concentrated and dried to a small volume. The samples were stored at 5 ℃.

Example 5

Fertilized egg isolate A

To produce fertilized egg isolate A, whole fertilized chicken eggs from 8 to 9 days old were sterilized with 70% ethanol and then placed in a fume hood to evaporate the solvent. The eggs were broken and the contents were dropped on, or through, a sterile 1.0mm screen. The egg shell and filtrate were discarded. The retentate, which contains the embryos, clear sacs and all or most of the ovalbumin and consists of solid and semi-solid and/or liquid fractions, is cooled on ice and then homogenized at 5 ℃. The homogenate (slurry) was poured into a sterile stainless steel tray and freeze-dried. The dried product was ground to a powder using a grinder to obtain isolate a. To isolate A, the preservatives sodium benzoate (0.5% w/w) and potassium sorbate (0.2% w/w) were added and the mixture was mixed. The final powder was stored at 2-8 ℃ (short term) or-20 ℃ (long term).

HPLC analysis

The final powder containing fertilized egg isolate a was analyzed by high performance (high pressure) liquid chromatography (HPLC). The results were quantified using a multi-wave absorption detector. The absorbance was read at 215 nm. Pharmacia Superdex 20010/300GL size exclusion column (10mm i.d.. times.300 mm) was used for fractionation. The separation range of the column is 10kDa to 600 kDa. The column was equilibrated with 20mM phosphate +0.3M NaCl pH 7.5. The sample was analyzed at a flow rate of 0.5 mL/min. Representative chromatograms are shown in fig. 6.

Verification of an analysis

The final powder containing isolate a was also analyzed through standard analytical procedures to measure purity and levels of protein, fat, ash, moisture and various contaminants. The results for representative samples are shown in fig. 7.

Preparation A capsule

To prepare a capsule of formulation A, 4000.0g (+/-2%) of the final powder containing isolate A, sodium benzoate (0.5% w/w) and potassium sorbate (0.2% w/w) were mixed with 40g (+/-2%) fumed silica using geometric dilution. The mixture was sieved and mixing and sieving were repeated to form formulation a. The formulation a mixture was encapsulated using Mini-Cap 300#0 white capsules to a target fill weight of 505mg to produce formulation a capsules.

Example 6

For the treatment of major depression of formulation ASymptoms of (1)(MDD) and investigation of its associated disorders/symptoms Is especially suitable for the treatment of diabetes

The efficacy and safety of fixed dose formulation a for the treatment of psychiatric disorders (e.g., MDD and related disorders and symptoms) was investigated. The study included an assessment of the effect of formulation a on reducing anxiety symptoms, improving quality of life, and improving symptoms of sexual dysfunction.

Description of evaluation techniques

Hamilton Depression rating Scale-17 item, "HAM-D" or "HAM-D17"

This is the most prominent rating scale used in north america for assessing patient depression. The total score can be interpreted as follows: very severe, > 23; severe, 19-22; medium, 14-18; slight, 8-13; and no depression, 0-7.

Hamilton anxiety rating Scale-14 item- "HAM-A"

The rating scale may assess the level of anxiety in the patient. The score levels are explained as follows: < 17, slight; 18-24, mild to moderate; and 25-30, moderate to severe.

Depression rating Scale, "MADRS"

This is the most prominent rating scale used in north america for assessing patient depression. According to the study, the following mean scores were associated with the overall severity measure: quite severe, 44; severe, 31; medium, 25; mild, 15; and rehabilitation, 7.

Beck Depression questionnaire- "BDI"

This is a commonly used measure of depression symptoms, and is commonly used as a self-assessment tool. The total score is a simple sum of the 21-term scores. Generally, scores < 9 indicate no or minimal depression, 10-18 indicate mild to moderate depression, 19-29 indicate moderate to severe depression, and > 30 indicates severe depression. However, scores of 0-4 may indicate a possible absence of depression, and scores of 40-63 may indicate a possible exaggeration of depression or personality disorders of the behavioural or borderline type.

Arizona sexual experience Scale, ASEX "

This is a 5-point rating scale that quantifies libido and assesses sexual arousal level, vaginal lubrication/penile erection, ability to achieve orgasm, and orgasm satisfaction.

The possible total score value ranges from 5 to 30, with higher scores indicating greater sexual dysfunction.

General health questionnaire score, GHQ "

The quality of the life dimension may be evaluated in a profile 36 (SF-36). The questionnaire can assess such problems as ability to concentrate attention, feelings of anxiety, poor confidence, poor self-value, worry, and depression. The scores were as follows:

the Likert scale (Likert scale), from left to right, 0, 1, 2, 3.

12 entries, each entry 0-3.

The scores ranged from 0 to 36.

Scores varied with study population. The score is typically about 11-12.

Score > 15, signs of distress.

Scores > 20 indicate severe problems and psychological stress.

Diagnostic and Statistical Manual of mental disorders Mental Disorders) -IV th edition-revised text- "DSM-IV TR"

This is the standard diagnostic manual for mental health professionals in north america, which comprehensively classifies mental disorders and provides widely accepted criteria for diagnosing mental disorders based on the best empirical evidence available.

The main effect of the measurement is analysis of variance of the repeated measurements and the score of HAM-D is used as the output variable. The second performance metrics include CGI-S and CGI-I, MADRS, SF36, BDI, HAMA, and ASEX.

Description of the study

An open study was conducted at the west neilsberg hospital (MSH) located in toronto, ontario, canada. Patients were recruited by media advertisements, referrals from MSH outpatient plans, and other clinical centers.

This protocol describes an open pilot study to investigate the potential antidepressant activity of formulation a. The objective of this preliminary study was to demonstrate that formulation a has the potential to improve MDD significantly above the level of known placebo effect recognized in other trials; and to demonstrate that formulation a is a useful therapy in this patient population. The second objective of this pilot study was to evaluate the effect of formulation a in reducing anxiety symptoms and improving quality of life.

Each patient was screened for MDD using DSM-IV TR standard and HAM-D. Once they participated in the study, they were scheduled into an open formulation a study over an 8-week period. The patients were further rated by global measurements, CGI severity (GCI-S) scale and CGI improvement (CGI-I) scale. The Udvalg for Kliniske is used for side effects(UKU) side effects rating Scale (Lingjaerde) was evaluated systematically. A second measure of depression symptoms is MontgomeryThe depression rating scale (MADRS) and Beck depression questionnaire (BDI) as a self-assessment tool. The quality of the life dimension may be evaluated in a profile 36 (SF-36). Anxiety Using 14 itemsHAM-A evaluation.

In a fixed dose open trial, patients are treated for depression based on a standard depression treatment regimen. Researchers determined the severity of depression on the rating scale at baseline times and at 2, 4, 6 and 8 weeks (W) of review. During the weeks of intervention, patients were observed with a short clinical assessment (V) to assess depression and drug tolerance.

Dosage of formulation A

The dose of formulation a was about 2000 mg/day (2 formulation a capsules, each about 500mg, taken twice daily).

Patient's inclusion criteria

To enroll in the study, the patient must meet a number of enrollment criteria, including criteria (i) - (vi) as described below.

(i) Clinical diagnosis meeting criteria for DSM-IV TR major depression, single episode or relapse.

(ii) The Hamilton Depression rating Scale (HAM-D17) with a total score of 18 or more for time base.

(iii)18-65 years old men/women who need new medication for diagnosed major depression or who need to change their existing medication for diagnosed major depression. Treatment decisions are made solely at the discretion of the clinician's discretion as appropriate to the patient's standard of care. However, no enhancement strategy was allowed during 8 weeks of the experiment.

(iv) English reading and writing ability.

(v) Obtaining an informed consent form of the written signature,

(vi) pregnancy tests were negative at screening.

Exclusion criteria

Patients were excluded from the study if they met a number of exclusion criteria, including the following criteria (i) - (xiii).

(i) Any other DSM IV TR diagnosis, including clinical diagnosis of depression other than DSM-IV TR MDD (excluding single episode/relapse, e.g., chronic depression and/or treatment-refractory depression).

(ii) Judged to be at significant suicide risk (HAMD suicide score > 1), or a history of impulsions (current potential) indicating a high desire for self-disability.

(iii) Any antidepressant other than formulation a.

(iv) Subjects who take and are unable or unwilling to stop a natural health product for depression.

(v) Pregnant, breast-fed, planned pregnancy in the next 12 months, or insufficiently protected women from contraception.

(vi) Clinically significant organ system diseases, such as cardiovascular diseases, liver diseases, kidney diseases, endocrinopathy, gastrointestinal diseases, metabolic diseases or other system diseases.

(vii) Electroconvulsive therapy (ECT) experience at the observation period.

(viii) Suffering from a major neurological disorder (i.e. parkinson's disease, huntington's disease), cerebrovascular disease (i.e. stroke), a metabolic disorder (i.e. vitamin B12 deficiency), an autoimmune disorder (i.e. systemic lupus erythematosus), a viral or other infection (i.e. hepatitis, mononucleosis, human immunodeficiency) or cancer.

(ix) Clinical or subclinical hypothyroidism/hyperthyroidism (e.g., elevated TSH).

(x) Allergic reactions to poultry or eggs.

(xi) Subjects who have been undergoing psychotherapy or who have been initiating psychotherapy in a trial.

(xii) Subjects with clinically significant abnormal laboratory results in blood and urine analysis were screened.

(xiii) Subjects that became significantly severe during the washout period (washout period).

Design of research

This was a single point, open randomized study in 25 patients designed to verify the efficacy and safety of formulation a monotherapy.

The trial consisted of an 8-week evaluation period, followed, if necessary, by a 2-week antidepressant clearance period.

Screening

Once the physician and/or study coordinator adequately advises the subject of the study, the nature of the treatment, and their other options, and the subject signs the informed consent text, the physician makes a clinical DSM IV TR diagnosis and administers HAM-D17. Qualified subjects were then reviewed for medical, psychiatric history, and concurrent therapy, followed by physical examination. In addition, study coordinator performed time-based laboratory tests including urine (Routine & microcopic), CBC classification (CBC differential), and platelets, electrolytes, bilirubin, BUN, creatinine, TSH, liver function tests, serum creatinine, and ECG. Pregnancy screening of female patients was obtained by hCG blood testing. Pregnant patients were excluded as well as those with clinically significant abnormalities in the laboratory tests.

Week 0

The patients were re-examined for baseline time (week 0) and scheduled by the physician to the formulation a monotherapy group. Patients who were depressed and were using antidepressants but had no effect were changed to formulation a.

The next few weeks

After initial evaluation and initiation of treatment with formulation a (V1 and V2), periodic weekly checks were continued for 8 weeks (W2-W8, V3-V6). Patients with other antidepressant drugs and selected for participation in the study entered a 1-2 week washout period and then started an 8 week active drug trial. The washout period is at the physician's clinical discretion. During this period, clearance of the patient was monitored by a psychiatric doctor once a week and further monitored by the study coordinator by telephone at mid-week intervals. It is recognized that depression worsens during the clearance period. However, if the previous drug was ineffective or partially ineffective, then a delay of 1-2 weeks would not significantly cause the risk of a depressive exacerbation (decline) to be significantly greater than with routine care in this regimen, provided that the subject is carefully monitored during this time and appropriate intervention is performed as needed. If formulation A is not an effective antidepressant for a particular patient, then that patient may be at risk for excessive prolongation of depression. However, depression is a chronic disorder that usually exists for months before being diagnosed or treated, and therefore an additional 8 weeks should be essentially indistinguishable from standard of care with careful monitoring and administration of formulation a, a potentially effective agent. Moreover, the standard of care already detailed is effective in only about 60% of patients, thus often requiring re-assessment and drug changes that are equally possible.

At V2 (possibly in combination with V1 (W0)) to V6(W8), the following steps are carried out by the mentoring Psychologist (PI) and/or research coordinator:

-body weight

-height of a person

-vital signs

Hamilton depression rating scale (item 17) (HAM-D17) (Hamilton 1967).

Clinical global impression (CGI-S, CGI-I) (Guy)

Depression Rating Scale (MADRS) (Montgomery)

-Beck depression questionnaire (BDI) (10)

Quality of life (SF-36) (Ware)

Hamilton anxiety rating scale (HAMA) (Hamilton1959)

-Udvalg for Kliniske Undersogelser (UKU) (Lingjaerde) (adverse event reporting) (except in V2)

Drug compliance (except at V2)

Study examinations estimated to take about 1 hour, except that the baseline time examination may take 2 hours.

If subjects become more depressed in the study, they are evaluated by the primary investigator to determine the best clinical approach. If deemed necessary, formulation A is discontinued in favor of another antidepressant treatment. This is a clinical decision made based solely on best practice for treatment of depression and the best clinical benefit of the patient.

Physicians and study coordinators are allowed to make general helpful contacts with the patient and the contacts are usually limited to answering questions about the patient's disease course and treatment. Formal psychological treatment is not allowed.

Statistical method

The primary effect was tested using repeated measures of analysis of variance with the score of HAM-D17 as the output variable. Significant temporal effects may support this assumption. The total expected sample size for 25 patients was large enough to detect HAM-D17 changes with a standard deviation of 0.65 as follows (single sample two-tailed P < 0.05). The reported standard deviation of HAM-D-17 is in the range of 4.5 to 6.5. Thus, the study design has 80% of the possible (power) to detect an average change on the 52-point table as small as 4.3 points. Each participant had a HAM-D17 score greater than 17 according to the addition criteria. The Franck mitigation criterion is a HAM-D17 of9 or less. The study used a more conservative and well-recognized level of 7 or less. An effect size of 4.3 is sensitive enough to detect clinical improvement from scores above 17 to scores below 10. Positive results are statistically based on the predicted placebo response rate in the depression treatment trial ranging from 30% to 50%. In this study, a placebo response rate of 40% was assumed. Some analysis was performed on responders and responders (remitter) as needed.

Results

A total of 23 patients participated in the study. 3 of these subjects (#104, #105, and #118) were never treated, and their results were therefore considered to be inseparable. Of the 20 subjects who received at least one dose of formulation a, 16 of them completed the 8-week study. The remaining 4 subjects did not complete the study for 8 weeks, but since they all received at least one dose of formulation a, their results were considered analyzable. Reasons for the incompletion of the 4 subjects for the entire study included non-compliance and/or non-compliance with the medication, non-patience with the results, and subject's departure from the country.

The results for 20 subjects receiving at least one dose of formulation a are provided in the table below.

Table of total scores of HAM-D

Screening	Week 0	Week 2	Week 4	Examination of No. 7	Week 6	Week 8
							Subject #101	20	14		11	7	3
Subject #102	19	15	5		2	0
							Subject #103	22	7	3		5	0
Subject #106	21	4	8		10	12
							Subject #107	22	17	20
Subject #108	20	14	19
							Subject #109	25	16	17		20	24
Subject #110	21	10	17		8	4
							Subject #111	24	20	19	19	23	23
Subject #112	29	8	5		2	0

Screening	Week 0	Week 2	Week 4	Examination of No. 7	Week 6	Week 8
							Subject #113	33	13	9		11	8
Subject #114	29	13	19		22	30
							Subject #115	32	8	13		5	6
Subject #116	19	17	24		17	24
							Subject #117	23	11	9		8
Subject #119	23	23	20		13	10
							Subject #120	23	5
Subject #121	23	11	8		6	3
							Subject #122	32	22	16		23	16
Subject #123	24	19	12		10	11

Total rating Table of GHQ

Screening	Week 0	Week 2	Week 4	Examination of No. 7	Week 6	Week 8
							Subject #101	15	27		14	11	3
Subject #102	22	11	3		0	0
							Subject #103	18	9	13		2	2
Subject #106	22	10	10		6	10
							Subject #107	27	12	9
Subject #108	27	19	17
							Subject #109	25	16	16		17	20
Subject #110	28	13	16		9	8
							Subject #111	26	15	20	21	21	20
Subject #112	30	15	9		4	0
							Subject #113	34	13	12		5	2
Subject #114	33	25	22		22	28
							Subject #115	31	2	7		5	8
Subject #116	32	26	25		25	24
							Subject #117	24	14	7		8
Subject #119	31	19	27		20	8
							Subject #120	23	8
Subject #121	35	7	1		1	1
							Subject #122	31	23	10		23	15
Subject #123	26	17	11		4	2

Total rating table for MADRS

Screening	Week 0	Week 2	Week 4	Examination of No. 7	Week 6	Week 8
							Subject #101	34	24		28	10	6
Subject #102	30	18	10		0	2
							Subject #103	28	10	4		2	2
Subject #106	30	14	14		18	24

Subject #107	38	28	28
							Subject #108	20	18	22
Subject #109	28	23	20		28	26
							Subject #110	28	16	36		14	10
Subject #111	46	40	40	40	32	36

Screening	Week 0	Week 2	Week 4	Examination of No. 7	Week 6	Week 8
							Subject #112	38	16	10		10	2
Subject #113	46	16	18		14	6
							Subject #114	42	26	38		38	44
Subject #115	32	12	12		10	10
							Subject #116	36	42	44		34	46
Subject #117	36	22	10		6
							Subject #119	38	34	34		18	10
Subject #120	32	6
							Subject #121	38	14	6		6	4
Subject #122	44	38	22		32	24
							Subject #123	30	28	22		14	16

Total score Table for BDI-21

Screening	Week 0	Week 2	Week 4	Examination of No. 7	Week 6	Week 8
							Subject #101	27	25		27	17	10
Subject #102	25	13	6		0	1
							Subject #103	26	14	10		8	8
Subject #106	30	12	7		12	40
							Subject #107	33	28	26
Subject #108	32	14	20
							Subject #109	29	23	24		20	25

Subject #110	29	22	24		13	8
							Subject #111	32	28	27	33	27	27
Subject #112	37	21	10		9	1
							Subject #113	53	23	22		18	3
Subject #114	54	40	52		52	59
							Subject #115	39	13	16		3	4
Subject #116	38	37	37		42	40
							Subject #117	24	20	11		7
Subject #119	35	36	40		24	18
							Subject #120	26	1
Subject #121	43	10	4		3	3
							Subject #122	55	38	25		46	27
Subject #123	33	34	22		5	10

Table of total scores of HAM-a

Screening	Week 0	Week 2	Week 4	Examination of No. 7	Week 6	Week 8
							Subject #101	15	9		8	7	3
Subject #102	13	8	2		0	1
							Subject #103	8	7	2		0	2
Subject #106	21	5	5		5	10
							Subject #107	12	7	13
Subject #108	15	8	11
							Subject #109	17	12	13		18	14
Subject #110	14	8	9		2	5
							Subject #111	33	29	24	32	23	24
Subject #112	22	10	4		1	2
							Subject #113	35	22	14		4	3

Subject #114	24	10	18		21	21
							Subject #115	25	10	12		4	5
Subject #116	12	13	15		12	16
							Subject #117	21	10	8		7
Subject #119	13	19	15		13	6
							Subject #120	14	4
Subject #121	24	9	3		4	2
							Subject #122	43	31	21		32	22
Subject #123	23	27	14		8	10

Total score table for ASEX

Screening	Week 0	Week 2	Week 4	Examination of No. 7	Week 6	Week 8
							Subject #101	12	14		12	13	13
Subject #102	21	23	19		19	15
							Subject #103	12	12	17		17	13
Subject #106	10	9	7		7	10
							Subject #107	15	14	16
Subject #108	15	11	12
							Subject #109	17	15	17		19	19
Subject #110	26	25	28		28	27
							Subject #111	28	28	28	28	28	28
Subject #112	11	13	11		11	9
							Subject #113	28	28	28		28	9
Subject #114	28	28	28		30	30

Subject #115	19	17	17		11	15
							Subject #116	11	10	11		12	12
Subject #117	14	12	18		16
							Subject #119	30	30	28		30	30
Subject #120	19	15
							Subject #121	18	26	12		22	10
Subject #122	26	14	15		30	12
							Subject #123	22	20	20		16	20

Response rate and intensity of response

The following definitions were used to assess the response of each subject to treatment with formulation a. By "responder" or "sustained responder" (or "sustained responder") is meant a subject who has at least a 50% improvement in the Hamilton depression rating scale (HAM-D score) compared to the baseline score at any time in the study. By "clinical responder" is meant a subject who meets the criteria for "responder" and who is considered by the primary investigator to have a positive clinical result. An "end of study responder" is a subject who meets responder criteria at the end of the study (or at the last observation). An "improvement" is a decrease in the HAM-D score to less than 8.

The above study showed that 15 of them (75%) were persistent responders and 14 of them (70%) were clinical responders to 20 subjects receiving at least one dose of formulation a. In addition, the number of persistent responders was 13/16 (81.3%) and the number of clinical responders was 12/16 (75%) among 16 subjects who completed the 8-week study. In addition, the overall reduction in HAM-D score (including non-responders) was significant (56.08%) in 16 subjects who completed the study. The reduction in HAM-D score was higher (68.1%) in the sustained responders who completed the 8 week study, a number much higher than the minimum 50% reduction required for sustained response.

It should be noted that the response of two subjects is influenced by the surrounding environment. Subject #114, who was not included in the clinical responder, was responsive to formulation a by week 2 when her HAM-D score decreased by more than 50%; but with the intervention of external factors. She began to encounter medical problems (unrelated to formulation a) and was difficult to work at which time she applied for incapacitance. These environmental factors completely overwhelm her favorable emotional response to formulation a.

Subject #106 should not be considered a responder at week 8 based on the strict criterion of 50% reduction in HAM-D score, since her score was 12 at this time, just not enough to be a 50% reduction from the score 21 when she participated in. However, subject #106 did respond throughout the 8 week trial: the score was 4 at week 2, 8 at week 4, and 10 at week 6. In fact, subject #106 was considered a clinical responder by PI during the Study and participated in the time delay Study (see example 7) where scores of 1, 11, 7 and 9 were recorded. After the start of the time-lapse study, subject #106 was faced with considerable household confusion, which undermined her positive response to formulation a. After the confusion subsided, she continued to maintain a response to formulation a. No drug can completely compensate for the traumatic effects of the surrounding environment. Formulation a has been good at improving the emotional trauma of these environments to subject # 106.

Good rate of conversion

Not all persistent responders, and not everyone who developed improvement, remained improved to the end of the 8 week study. Improvement occurred in 9 of the 15 persistent responders (60%) at some point during the 8 week study. 7 of these 9 patients who improved (77.8%; or 46.7% of all study participants) were in remission by the end of the 8 week study.

The following table depicts all study participants who experienced improvement and all study participants who remained improved. The number on line indicates that the subject had or had been in remission, while the number X indicates that the subject did not have or did not remain in remission until week 8 of the study.

In addition, all but 1 persistent responder presented another major outcome, anxiety reduction. These results indicate that formulation a is effective in treating major depression and anxiety. Furthermore, there were no serious side effects caused by this drug. In the subjects involved in the study, there was no weight gain, and there was also a loss of asexual function.

Example 7

The positive efficacy and safety results of the study described in example 6 require a time-lapse study. 10 subjects from the study described in example 6 participated in the time-lapse study. The time-lapse study was open only to those subjects in the study described in example 6 who were clinical responders at the end of the 8-week study. Formulation a was administered as described in example 6, and subjects from the time-lapse study were analyzed monthly for 10 months. The following table shows the HAM-D scores of subjects in the time-lapse study.

w/d-Exit delay study

4 of the 10 subjects dropped out of the time-lapse study due to the appearance of exclusion criteria for continued study. The results of this time-lapse study indicated that all subjects in the study were necessarily responders to formulation a. 6 of 10 clinical responders (60%) were in remission at the start of the delayed study. 8 of 10 subjects (80%) were in remission on the last day of assessment. 2 of the subjects were clinical responders in the initial 8-week study, but no improvement occurred until the end of the time-lapse study. Only 1 subject (#113) enrolled in the time-lapse study, who was a clinical responder, had relapsed of old disease after enrolled in the time-lapse study. Example 6 positive efficacy and safety results in the time-delayed study further time-delayed studies were required.

Example 8

As described in the above examples, formulation a has been shown to have a therapeutic effect. Studies have been conducted to investigate the mechanism of action of formulation a. In particular, studies were conducted to determine the inhibition of the binding interaction of the radioligand with its receptor or the action of radiolabeled enzymes on their associated target proteins by agent a. The level of inhibition of agent a (measured as percent inhibition of specific binding to each receptor by agent a) was determined. Inhibition of binding interaction and enzyme activity was tested at two different concentrations of formulation A (1.0. mu.g/mL and 10.0. mu.g/mL), which were repeated twice for each sample. These concentrations of formulation a can be prepared by: the capsule contents of formulation A were dissolved in dimethyl sulfoxide and the solution was subsequently diluted to 1.0. mu.g/mL or 10.0. mu.g/mL of formulation A. These diluted solutions are referred to as isolate a. Radioligand binding assays were then performed using the receptors and enzymes listed in the table below. The mean percent inhibition of specific binding was determined for each isolate a concentration. The results of this study are shown in the table below.

Generally, an inhibition of binding or inhibition of enzyme activity of 20% or less indicates that a given test compound is inactive for that particular receptor binding site or target protein. Greater than 20% inhibition of binding or enzyme activity indicates that the test compound is active on that particular receptor binding site or enzyme site. The binding inhibition studies described above indicated that isolate a replaced glutamate from the 4 major ionotropic receptors for glutamate. Binding of radiolabeled AMPA to the AMPA receptor was inhibited by 29.05% in the presence of isolate A (10. mu.g/mL). Binding of radiolabeled kainic acid to the kainic acid receptor was inhibited by 22.38% in the presence of isolate A (10. mu.g/mL). Binding of radiolabeled CGP 39653 to the agonist site of the NMDA receptor was inhibited by 34.59% in the presence of isolate A (10. mu.g/mL). Binding of radiolabeled MDL-105, 519 to the strychnine-insensitive glycine site of the NMDA receptor was inhibited by 27.45% in the presence of isolate A (10. mu.g/mL). In addition, binding of neurokinin A to the NK2 receptor was inhibited by 32.15% in the presence of isolate A (10. mu.g/mL). Additional details regarding binding inhibition assays for each of the 5 above receptors are provided in the table below.

The ionotropic glutamate receptor, NK2 receptor and neurokinin 1(NK1) receptor were used in additional receptor binding assays. A concentration-controlled experiment was performed to assess the ability of various isolates of the contents of formulation a capsules to antagonize ligand binding of various glutamate receptors and NK2 receptors. AMPA receptors, kainic acid receptors, agonists of NMDA receptors and glycine (strychnine insensitive) binding sites and NK2 receptors were studied in this assay. The contents of formulation a capsules were dissolved using a variety of solvents and extracted using 4 different methods, as detailed below. These extraction steps produce a variety of fertilized egg isolates. These isolates were called: sample #19 upper isolate, sample #19 lower isolate, sample #20 upper isolate, sample #20 lower isolate, fraction X isolate, and sample #2 isolate. Each of these isolates was then tested in a radioligand binding assay.

Sample #19 was prepared by weighing 103mg of the contents of formulation a capsules. Water (10.3mL) was added and the solution was vortexed for 1 min. Then 30mL of ethyl acetate were added to the solution and the solution was vortexed again for 1 minute. The samples were then centrifuged using a Beckman bench top centrifuge. As a result, 3 fractions were formed. The upper (organic) and lower (aqueous) fractions were collected separately and the middle fraction was discarded. Both the upper and lower fractions were completely dried. The lower fraction (aqueous phase) was redissolved in 2.06mL of water. The sample was not clarified and centrifuged at 10,000rpm for 10 minutes using a microcentrifuge. The supernatant was removed and labeled as sample 085426-4 (sample #19 lower isolate) and used in receptor binding studies. The upper fraction (organic phase) was redissolved in 1.245mL of 20% aqueous acetonitrile. The sample was not clarified and centrifuged at 10,000rpm for 10 minutes using a microcentrifuge. The supernatant was removed and labeled as sample 085426-3 (sample #19 supernatant) and used in receptor binding studies. A control of sample #19 was also prepared. This control consisted of a 20% aqueous solution of acetonitrile and was labeled as sample 085426-5 in the receptor binding study.

Sample #20 was prepared by weighing 249.7mg of the contents of formulation a capsules. 10mL of methanol: dichloromethane (1: 1) was added and the solution was vortexed. 10mL of dichloromethane was then added to the solution, and the solution was vortexed again. The samples were then centrifuged using a Beckman bench top centrifuge at 3500rpm for 15 minutes. As a result, 3 fractions were formed. The upper and lower organic fractions were collected separately. The intermediate fraction was discarded. Both the upper and lower fractions were completely dried and redissolved in 2.49mL of aqueous methanol (100%). The upper methanol fraction was semi-clear and the lower methylene chloride fraction was insoluble. Both samples were centrifuged at 10,000rpm for 10 minutes using a microcentrifuge. The supernatant of each sample was removed. The supernatant of the upper methanol fraction was labeled sample 085426-6 (sample #20 upper isolate) and used in receptor binding studies. The supernatant of the lower methylene chloride fraction was labeled sample 085426-7 (sample #20 lower isolate) and used in the receptor binding study. A control of sample #29 was also prepared. This control consisted of 10% aqueous methanol and was labeled as sample 085426-9 in the receptor binding study.

Sample fraction X was prepared as follows. The contents of 121mg of formulation A capsules were weighed out. Then 10mL of water was added. 10mL of dichloromethane was then added to the solution and the sample vortexed. The aqueous and organic fractions were each removed separately. The solvent separation was repeated by: to the aqueous fraction was added 10mL of dichloromethane and the solution was vortexed. The aqueous and organic fractions were again removed separately. The organic fractions from the two separations were combined and the aqueous fractions from the two separations were combined. The aqueous and organic fractions were completely dried and weighed. The aqueous fraction weighed 116.4mg and the organic fraction weighed 1.3 mg. The organic fraction was redissolved in 1.3mL of 10% aqueous methanol (at a concentration of 0.1mg/mL), labeled sample 085426-8 (fraction X isolate) and used in binding studies. A control of sample fraction X was also prepared. This control consisted of 10% aqueous methanol and was labeled as sample 085426-9 in the receptor binding study (note that this was the same control used for sample # 20).

Sample #2 was prepared as follows. One part of the contents of formulation a capsule (1.8mg) was weighed out. Then, 40% aqueous PEG solution (3.6mL) added 0.25% Tween 80 (0.5 mg/mL) was added and the sample was vortexed. This preparation was labeled as sample 085426-1 (sample #2 isolate) and tested in a receptor binding study. A control of sample #2 was also prepared. The control consisted of 40% aqueous PEG solution with 0.25% Tween 80 added and was labeled as sample 085426-2 in the receptor binding study.

The results of the receptor binding studies (two samples obtained from the maximum concentration of each isolate) are shown in the table below.

Bold indicates over 50% inhibition at the tested concentration.

Binding of radiolabeled AMPA to AMPA receptor was inhibited by 97% in the presence of the upper isolate of sample # 20. Binding of radiolabeled kainic acid to the kainic acid receptor was inhibited by 101% in the presence of the upper isolate of sample # 20. Binding of radiolabeled CGP 39653 to the agonist site of the NMDA receptor was inhibited by 110% in the presence of sample #20 supernatant isolate. Binding of radiolabeled MDL-105, 519 to the strychnine-insensitive glycine site of the NMDA receptor was inhibited by 92% in the presence of sample #20 upper isolate. In addition, fraction X isolate inhibited neurokinin a binding to its NK2 receptor by 55%. To the best of the inventors' knowledge, this is the first demonstration that the glutamate receptor and the NK2 receptor are antagonized by a single agent.

To further confirm that the binding and activation of the above glutamate receptor and the binding and activation of NK2 receptor were antagonized by the sample #20 supernatant isolate prepared as described above, dose-effect studies were performed. Inhibition of binding to AMPA receptor, kainate receptor, agonist site of NMDA receptor, glycine site of NMDA receptor (strychnine insensitive) and NK2 receptor was assessed in the presence of #20 supernatant isolate (based on the amount of formulation a contained in sample #20 prior to extraction) at the following concentrations: 0.1, 0.3, 1.0, 3.0, 10, 30, 100 and 300. mu.g/mL.

FIG. 8 shows knots determined with agonist sites of NMDA receptor (ionotropic)And (5) fruit. The ability of CGP 39653 to bind to the agonist site of the NMDA receptor was inhibited in a concentration-dependent manner by the upper isolate of sample #20, with higher concentrations of the upper isolate of sample #20 providing greater binding inhibition and lower concentrations providing less binding inhibition. IC of NMDA₅₀Was determined to be 1.11X 10^-5μg/mL，K_iWas determined to be 9.02X 10^-6And M. IC of supernatant isolate of sample #20₅₀Determined as 5.78. mu.g/mL, K_iIt was determined to be 4.69M.

FIG. 9 shows the results of an assay with the kainic acid receptor. The supernatant isolate of sample #20 inhibited the ability of kainic acid to bind to the kainic acid receptor in a concentration dependent manner, with higher concentrations of the supernatant isolate of sample #20 providing greater binding inhibition and lower concentrations providing less binding inhibition. IC of kainic acid₅₀Was determined to be 1.77X 10^-8μg/mL，K_iDetermined as 1.08X 10^-8And M. IC of supernatant isolate of sample #20₅₀Determined as 12.0. mu.g/mL, K_iThe determination was 7.32M.

FIG. 10 shows the results of the assay with the AMPA receptor. The supernatant isolate of sample #20 inhibited the ability of AMPA to bind to AMPA receptors in a concentration-dependent manner, with higher concentrations of the supernatant isolate of sample #20 providing greater inhibition of binding and lower concentrations providing less inhibition of binding. IC of (+/-) AMPA HBr₅₀Was determined to be 3.54X 10^-8μg/mL，K_iWas determined to be 3.00X 10^-8And M. IC of supernatant isolate of sample #20₅₀Determined as 11.7. mu.g/mL, K_iThe determination was 9.91M.

FIG. 11 shows the results of an assay with the (strychnine insensitive) glycine site of the NMDA receptor. The ability of the sample #20 supernatant isolate to bind the (strychnine insensitive) glycine site of the NMDA receptor was inhibited in a concentration dependent manner by the MDL-105, 519, with higher concentrations of sample #20 supernatant isolate providing greater inhibition of binding and lower concentrations providing less inhibition of binding. IC of MDL-105, 519₅₀Determined as 2.8X 10^-8μg/mL，K_iDetermined as 2.36X 10^-8And M. Sample #20IC of layer separation₅₀Determined as 21.9. mu.g/mL, K_iWas determined to be 18.4M.

FIG. 12 shows the results of the assay with NK2 receptor. Sample #20 supernatant inhibited the ability of neurokinin a to bind to the NK2 receptor in a concentration-dependent manner, with higher concentrations of sample #20 supernatant providing greater binding inhibition and lower concentrations providing less binding inhibition. IC of neurokinin A₅₀Was determined to be 6.84X 10^-10μg/mL，K_iDetermined as 5.76X 10^-10And M. IC of supernatant isolate of sample #20₅₀Was determined to be 4.15X 10²μg/mL，K_iWas determined to be 3.49X 10²M。

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Claims (79)

1. A method of producing fertilized egg isolate, said isolate comprising an embryo and a majority of ovalbumin and clear sac, said method comprising the steps of:

(i) optionally sterilizing the egg shell;

(ii) substantially separating the embryo, ovalbumin, and clear sac from the egg shell and other egg contents; and

(iii) freeze-drying the separated embryos, ovalbumin and clear sacs.

2. The method of claim 1, wherein the embryo, ovalbumin, and clear sac are separated from a substantial amount of the yolk.

3. The method according to claim 1, wherein the embryo, ovalbumin, and clear sac are separated by subjecting the contents of the fertilized egg to a filtration step.

4. The method of claim 3, wherein said filtering step uses a screen.

5. The method of claim 4, wherein the screen is a 1mm screen.

6. A fertilized egg isolate prepared according to the method of claim 1.

7. A composition of fertilized egg isolate, said composition comprising fertilized egg isolate produced according to claim 3 and a pharmaceutically acceptable carrier.

8. The composition of claim 7, wherein the support is fumed silica.

9. The composition of claim 7, further comprising one or more preservatives.

10. The composition of claim 9, wherein the one or more preservatives are selected from sodium benzoate and potassium sorbate.

11. A dosage form comprising the fertilized egg isolate of claim 6.

12. The dosage form of claim 11, wherein the dosage form is a tablet or capsule.

13. A capsule comprising the fertilized egg isolate of claim 6.

14. A method of treating depression using fertilized egg isolate, said method comprising the steps of: administering a therapeutically effective amount of the fertilized egg isolate to a patient in need thereof.

15. The method according to claim 14, wherein the fertilized egg isolate comprises freeze-dried embryo, ovalbumin, and clear sac from fertilized eggs.

16. The method according to claim 15, wherein the fertilized egg isolate does not contain a substantial amount of yolk from the fertilized egg.

17. The method of claim 14, wherein the patient is not treated by psychotherapy concurrently with the treatment.

18. The method of claim 14, wherein the patient is treated by psychotherapy concurrently with the treatment.

19. A method of treating depression using the fertilized egg isolate produced according to claim 6, the method comprising the steps of: administering a therapeutically effective amount of the fertilized egg isolate to a patient in need thereof.

20. A method of treating major depressive disorder using fertilized egg isolate, the method comprising the steps of: administering a therapeutically effective amount of the fertilized egg isolate to a patient in need thereof.

21. The method according to claim 20, wherein the fertilized egg isolate comprises freeze-dried embryo, ovalbumin, and clear sac from the fertilized egg.

22. The method according to claim 21, wherein the fertilized egg isolate does not contain a substantial amount of yolk from the fertilized egg.

23. The method according to claim 20, wherein the patient is not treated by psychotherapy concurrently with the treatment.

24. The method according to claim 20, wherein the patient is treated by psychotherapy concurrently with the treatment.

25. A method of treating major depressive disorder using the fertilized egg isolate produced according to claim 6, the method comprising the steps of: administering a therapeutically effective amount of the fertilized egg isolate to a patient in need thereof.

26. A method of treating anxiety using fertilized egg isolate, said method comprising the steps of: administering a therapeutically effective amount of the fertilized egg isolate to a patient in need thereof.

27. The method according to claim 26, wherein the fertilized egg isolate comprises freeze-dried embryo, ovalbumin, and clear sac from the fertilized egg.

28. The method according to claim 27, wherein the fertilized egg isolate does not contain a substantial amount of yolk from the fertilized egg.

29. The method of claim 26, wherein the patient is not treated by psychotherapy concurrently with the treatment.

30. The method of claim 26, wherein the patient is treated by psychotherapy concurrently with the treatment.

31. A method of treating anxiety using fertilized egg isolate produced according to claim 6, the method comprising the steps of: administering a therapeutically effective amount of the fertilized egg isolate to a patient in need thereof.

32. A method of treating sexual dysfunction using fertilized egg isolate, said method comprising the steps of: administering a therapeutically effective amount of the fertilized egg isolate to a patient in need thereof.

33. The method according to claim 32, wherein the fertilized egg isolate comprises freeze-dried embryo, ovalbumin, and clear sac from the fertilized egg.

34. The method according to claim 33, wherein the fertilized egg isolate does not contain a substantial amount of yolk from the fertilized egg.

35. The method of claim 32, wherein the patient is not treated by psychotherapy concurrently with the treatment.

36. The method of claim 32, wherein the patient is treated by psychotherapy concurrently with the treatment.

37. A method of treating sexual dysfunction using fertilized egg isolate produced according to claim 6, the method comprising the steps of: administering a therapeutically effective amount of the fertilized egg isolate to a patient in need thereof.

38. A method of treating a disorder selected from the group consisting of a depressive affective disorder and an anxiety disorder using fertilized egg isolate, the method comprising the steps of: administering a therapeutically effective amount of the fertilized egg isolate to a patient in need thereof.

39. The method according to claim 38, wherein the fertilized egg isolate comprises freeze-dried embryo, ovalbumin, and clear sac from the fertilized egg.

40. The method according to claim 39, wherein the fertilized egg isolate does not contain a substantial amount of yolk from the fertilized egg.

41. The method of claim 38, wherein the patient is not treated by psychotherapy concurrently with the treatment.

42. The method of claim 38, wherein the patient is treated by psychotherapy concurrently with the treatment.

43. A method of treating a disorder selected from the group consisting of depressive affective disorder and anxiety disorder using fertilized egg isolate produced according to claim 6, the method comprising the steps of: administering a therapeutically effective amount of the fertilized egg isolate to a patient in need thereof.

44. A method of treating depression using an aqueous fertilized egg isolate, the method comprising the steps of: administering to a patient in need thereof a therapeutically effective amount of the aqueous fertilized egg isolate produced by a process comprising the steps of: (i) substantially separating the contents of the at least one fertilized egg from the egg shell; (ii) optionally filtering the contents to produce a retentate; (iii) combining the contents or retentate in a container; (iv) optionally adding a solvent, mixing the contents or retentate to produce a slurry; (v) mixing the slurry with an aqueous solution for a period of time; (vi) clarifying the mixture to produce an aqueous solution substantially free of solid matter; (vii) separating the aqueous solution from the solid material; (viii) freezing the aqueous solution; and (ix) freeze drying the aqueous solution to produce the fertilized egg isolate.

45. A method of treating anxiety using an aqueous fertilized egg isolate, the method comprising the steps of: administering to a patient in need thereof a therapeutically effective amount of the aqueous fertilized egg isolate produced by a process comprising the steps of: (i) substantially separating the contents of the at least one fertilized egg from the egg shell; (ii) optionally filtering the contents to produce a retentate; (iii) combining the contents or retentate in a container; (iv) optionally adding a solvent, mixing the contents or retentate to produce a slurry; (v) mixing the slurry with an aqueous solution for a period of time; (vi) clarifying the mixture to produce an aqueous solution substantially free of solid matter; (vii) separating the aqueous solution from the solid material; (viii) freezing the aqueous solution; and (ix) freeze drying the aqueous solution to produce the fertilized egg isolate.

46. A method of treating sexual dysfunction using aqueous fertilized egg isolate, said method comprising the steps of: administering to a patient in need thereof a therapeutically effective amount of the aqueous fertilized egg isolate produced by a process comprising the steps of: (i) substantially separating the contents of the at least one fertilized egg from the egg shell; (ii) optionally filtering the contents to produce a retentate; (iii) combining the contents or retentate in a container; (iv) optionally adding a solvent, mixing the contents or retentate to produce a slurry; (v) mixing the slurry with an aqueous solution for a period of time; (vi) clarifying the mixture to produce an aqueous solution substantially free of solid matter; (vii) separating the aqueous solution from the solid material; (viii) freezing the aqueous solution; and (ix) freeze-drying the aqueous solution to produce the fertilized egg aqueous isolate.

47. A method of treating a disorder selected from the group consisting of depressive affective disorder and anxiety disorder using an aqueous fertilized egg isolate, and comprising the steps of: administering to a patient in need thereof a therapeutically effective amount of the aqueous fertilized egg isolate produced by a process comprising the steps of: (i) substantially separating the contents of the at least one fertilized egg from the egg shell; (ii) optionally filtering the contents to produce a retentate; (iii) combining the contents or retentate in a container; (iv) optionally adding a solvent, mixing the contents or retentate to produce a slurry; (v) mixing the slurry with an aqueous solution for a period of time; (vi) clarifying the mixture to produce an aqueous solution substantially free of solid matter; (vii) separating the aqueous solution from the solid material; (viii) freezing the aqueous solution; and (ix) freeze-drying the aqueous solution to produce the fertilized egg aqueous isolate.

48. A method of treating depression using fertilized egg solvent isolate, said method comprising the steps of: administering a therapeutically effective amount of the fertilized egg solvent isolate to a patient in need thereof, the fertilized egg isolate being produced by a process comprising the steps of: (i) substantially separating the contents of the at least one fertilized egg from the egg shell; (ii) optionally filtering the contents to produce a retentate; (iii) combining the contents or retentate in a container; (iv) optionally adding a solvent, mixing the contents or retentate to produce a slurry; (v) mixing the slurry with a hydrophobic solvent for a period of time; (vi) clarifying the mixture to produce a substantially solid matter free solution; (vii) separating the solution from the solid material; (viii) separating the hydrophobic solvent portion from any aqueous solvent portion; and (ix) concentrating the hydrophobic solvent fraction to produce a fertilized egg solvent isolate.

49. A method of treating anxiety using fertilized egg solvent isolate, said method comprising the steps of: administering a therapeutically effective amount of the fertilized egg solvent isolate to a patient in need thereof, the fertilized egg isolate being produced by a process comprising the steps of: (i) substantially separating the contents of the at least one fertilized egg from the egg shell; (ii) optionally filtering the contents to produce a retentate; (iii) combining the contents or retentate in a container; (iv) optionally adding a solvent, mixing the contents or retentate to produce a slurry; (v) mixing the slurry with a hydrophobic solvent for a period of time; (vi) clarifying the mixture to produce a substantially solid matter free solution; (vii) separating the solution from the solid material; (viii) separating the hydrophobic solvent portion from any aqueous solvent portion; and (ix) concentrating the hydrophobic solvent fraction to produce a fertilized egg solvent isolate.

50. A method of treating sexual dysfunction using fertilized egg solvent isolate, said method comprising the steps of: administering a therapeutically effective amount of the fertilized egg solvent isolate to a patient in need thereof, the fertilized egg isolate being produced by a process comprising the steps of: (i) substantially separating the contents of the at least one fertilized egg from the egg shell; (ii) optionally filtering the contents to produce a retentate; (iii) combining the contents or retentate in a container; (iv) optionally adding a solvent, mixing the contents or retentate to produce a slurry; (v) mixing the slurry with a hydrophobic solvent for a period of time; (vi) clarifying the mixture to produce a substantially solid matter free solution; (vii) separating the solution from the solid material; (viii) separating the hydrophobic solvent portion from any aqueous solvent portion; and (ix) concentrating the hydrophobic solvent fraction to produce a fertilized egg solvent isolate.

51. A method of treating a disorder selected from the group consisting of depressive affective disorder and anxiety disorder using fertilized egg solvent isolate, and the method comprising the steps of: administering a therapeutically effective amount of the fertilized egg solvent isolate to a patient in need thereof, the fertilized egg isolate being produced by a process comprising the steps of: (i) substantially separating the contents of the at least one fertilized egg from the egg shell; (ii) optionally filtering the contents to produce a retentate; (iii) combining the contents or retentate in a container; (iv) optionally adding a solvent, mixing the contents or retentate to produce a slurry; (v) mixing the slurry with a hydrophobic solvent for a period of time; (vi) clarifying the mixture to produce a substantially solid matter free solution; (vii) separating the solution from the solid material; (viii) separating the hydrophobic solvent portion from any aqueous solvent portion; and (ix) concentrating the hydrophobic solvent fraction to produce a fertilized egg solvent isolate.

52. A method of treating depression using fertilized egg isolate, said method comprising the steps of: administering a therapeutically effective amount of the fertilized egg isolate to a patient in need thereof, the fertilized egg isolate being produced by a method comprising the steps of: (i) substantially isolating at least one embryo from the at least one fertilized egg; (ii) optionally washing the at least one embryo; (iii) freezing the at least one embryo; (iv) freeze-drying the at least one frozen embryo; and (v) comminuting the at least one freeze-dried embryo to form a fertilized egg isolate.

53. A method of treating anxiety using fertilized egg isolate, said method comprising the steps of: administering a therapeutically effective amount of the fertilized egg isolate to a patient in need thereof, the fertilized egg isolate being produced by a method comprising the steps of: (i) substantially isolating at least one embryo from the at least one fertilized egg; (ii) optionally washing the at least one embryo; (iii) freezing the at least one embryo; (iv) freeze-drying the at least one frozen embryo; and (v) comminuting the at least one freeze-dried embryo to form a fertilized egg isolate.

54. A method of treating sexual dysfunction using fertilized egg isolate, said method comprising the steps of: administering a therapeutically effective amount of the fertilized egg isolate to a patient in need thereof, the fertilized egg isolate being produced by a method comprising the steps of: (i) substantially isolating at least one embryo from the at least one fertilized egg; (ii) optionally washing the at least one embryo; (iii) freezing the at least one embryo; (iv) freeze-drying the at least one frozen embryo; and (v) comminuting the at least one freeze-dried embryo to form a fertilized egg isolate.

55. A method of treating a disorder selected from the group consisting of a depressive affective disorder and an anxiety disorder using fertilized egg isolate, the method comprising the steps of: administering a therapeutically effective amount of the fertilized egg isolate to a patient in need thereof, the fertilized egg isolate being produced by a method comprising the steps of: (i) substantially isolating at least one embryo from the at least one fertilized egg; (ii) optionally washing the at least one embryo; (iii) freezing the at least one embryo; (iv) freeze-drying the at least one frozen embryo; and (v) comminuting the at least one freeze-dried embryo to form a fertilized egg isolate.

56. A method of using fertilized egg isolate to treat a disease or disorder associated with a glutamate receptor, said method comprising the steps of: administering a therapeutically effective amount of the fertilized egg isolate to a patient in need thereof.

57. The method of claim 56, wherein said glutamate receptor is an AMPA receptor, a kainate receptor, or an NMDA receptor.

58. The method of claim 56, wherein the disease or disorder associated with the glutamate receptor is depression, major depression, anxiety, Alzheimer's disease, epilepsy, schizophrenia, impairment of brain cell function following stroke/ischemia, amyotrophic lateral sclerosis (Lou Gehrig's disease), lathyrism, autism, mental retardation, cognitive disorders, bipolar depression, or mania.

59. The method according to claim 56, wherein the fertilized egg isolate comprises freeze-dried embryo, ovalbumin, and clear sac from the fertilized egg.

60. The method according to claim 59, wherein the fertilized egg isolate does not contain a substantial amount of yolk from the fertilized egg.

61. The method of claim 56, wherein the patient is not treated by psychotherapy concurrently with the treatment.

62. The method of claim 56, wherein the patient is treated by psychotherapy concurrently with the treatment.

63. A method of treating a disease or disorder associated with a glutamate receptor using the fertilized egg isolate produced according to claim 6, the method comprising the steps of: administering a therapeutically effective amount of the fertilized egg isolate to a patient in need thereof.

64. A method of treating a disease or disorder associated with the neurokinin 2(NK2) receptor using fertilized egg isolate, said method comprising the steps of: administering a therapeutically effective amount of the fertilized egg isolate to a patient in need thereof.

65. The method of claim 64, wherein said disease or disorder associated with said NK2 receptor is depression, anxiety, irritable bowel syndrome, inflammatory airway disease, or urinary incontinence.

66. The method according to claim 64, wherein the fertilized egg isolate comprises freeze-dried embryo, ovalbumin, and clear sac from the fertilized egg.

67. The method according to claim 66, wherein the fertilized egg isolate does not contain a substantial amount of yolk from the fertilized egg.

68. The method of claim 64, wherein the patient is not treated by psychotherapy concurrently with the treatment.

69. The method of claim 64, wherein the patient is treated by psychotherapy concurrently with the treatment.

70. A method of treating a disease or disorder associated with the NK2 receptor using the fertilized egg isolate produced according to claim 6, the method comprising the steps of: administering a therapeutically effective amount of the fertilized egg isolate to a patient in need thereof.

71. A method of inhibiting glutamate receptor activity comprising contacting said glutamate receptor with an effective amount of fertilized egg isolate.

72. The method of claim 71, wherein said glutamate receptor is an AMPA receptor, a kainate receptor, or an NMDA receptor.

73. The method of claim 71, wherein the method is an in vitro method.

74. The method of claim 71, wherein the method is an in vivo method.

75. The method according to claim 71, wherein the fertilized egg isolate is produced by the method of claim 6.

76. A method of inhibiting NK2 receptor activity, comprising contacting the NK2 receptor with an effective amount of fertilized egg isolate.

77. The method of claim 76, wherein the method is an in vitro method.

78. The method of claim 76, wherein the method is an in vivo method.

79. The method according to claim 76, wherein the fertilized egg isolate is produced by the method of claim 6.