HK1177391A - Compounds for treating disorders or diseases associated with neurokinin 2 receptor activity - Google Patents

Description

compounds for the treatment of disorders or diseases associated with neurokinin 2 receptor activity

Cross reference to related applications

This application claims the benefit of U.S. provisional application No.61/240,014, filed on 9,4, 2009, which is incorporated herein by reference.

Technical Field

The present invention relates to methods for the treatment and prevention of neurokinin 2 (NK)₂) Compounds, pharmaceutical compositions and methods of disorders or diseases associated with receptor activity.

Background

Depressive mood disorder

Depressive mood disorders are a group of mood disorders characterized by mood depression. Depressive mood disorders include major depressive disorder, dysthymic disorder, bipolar disorder, depression caused by general medical conditions such as depression associated with dementia or schizoaffective disorder, drug-induced depression, postpartum depression, and seasonal affective disorder.

Major depressive disorders (also known as major depression, clinical depression, unipolar depression, and unipolar disorder) are common in the general population. Recent North American data show 14.5% of adults at life-long risk of major depression and an annual prevalence of 8.1% (results from the 2004National surface on Drug Use and Health: National definitions; reviews as of 9/8/2005; Department of Health and Human Services. Substance Absuse and Mental Health Services Administration of Applied students).

Under modern therapy, the mean duration of a depressive episode 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 the age of antidepressant treatment (Kendler, McLeod, Patten).

Antidepressants have some effect in the treatment of major depressive disorders and in alleviating patient suffering. Not all actions are positive. The functioning of patients with major depressive disorders is often impaired and often accompanied by other unhealthy conditions, such as drug abuse, possibly due to a potential major depressive disorder. Major depressive disorders result in increased use of health services and can have devastating effects on community structure and socioeconomic performance.

The cause of major depressive disorder is not fully understood. Dysregulation of monoamine synthesis and activity has been the leading etiological theory of major depressive disorders for the past decades, and the support for this notion is reinforced by the efficacy of pharmacological therapies that enhance monoamine activity, particularly those of 5-hydroxytryptamine and/or norepinephrine. However, any given antidepressant is effective only in a small group of depressed patients, and often only partially. Current therapies performed in investigatively 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 major depressive disorders that indicate more complex interactions of the causative factors (including the role of second messengers in mediating membrane binding and intracellular processes). This led to the study of the following events: hormone pathways, such as: hypothalamus-pituitary-adrenal (HPA) axis (increased activity in 20-40% of community-inhabited patients with major depressive disorder), thyroid cervical stem (5-10% of patients assessed for major depressive disorder have not previously detected thyroid dysfunction), growth hormone, prolactin, testosterone; and inflammatory processes and their markers (e.g., interleukin 1, interleukin 6, and tumor necrosis factor).

Most people with major depressive disorders 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 rehabilitate and prevent relapse. 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 a sufficient drug dose for a sufficient duration of time for the patient, 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 gastrointestinal discomfort, headache, sleep disturbances, significant impairment of sexual function, and many other side effects. Most antidepressants have at least some significant side effects that limit the ability of the clinician to effectively treat many patients.

Major depressive disorders may be associated with other disorders and/or syndromes, including disorders of the brain or nervous system, anxiety disorders (including generalized anxiety disorder, panic disorder, phobias, obsessive compulsive disorder, post traumatic stress disorder, separation anxiety disorder, social anxiety disorder (also known as social phobia), bipolar disorder, and dementia); sexual dysfunction, drug abuse, eating disorders and hormonal disorders, such as thyroid dysfunction, hypogonadism, menopause, and the like. Treatment of major depressive disorders often results in an improvement of these related conditions and syndromes.

In addition, some therapeutic agents used to treat depression are also effective for treating other conditions. For example, antidepressants have been shown to be effective in treating hot flashes (hot flashes) associated with menopause, pain, and smoking cessation.

Anxiety disorder

Anxiety disorders are a group of disorders that affect behavior, thought, mood and physical health. Anxiety disorders are believed to be caused by a combination of biological factors and the individual's personal environment. People with anxiety disorders experience intense and prolonged panic and distress for unknown reasons. This disorder brings their lives into a continuous restless and fear history and may interfere with their relationships with family, friends and colleagues.

Anxiety disorders are the most common of all mental health problems. It is estimated to affect approximately one-tenth of a person. The proportion of women is greater than men, affecting not only adults but also children. People often suffer from more than one type of anxiety disorder in the category of anxiety disorders, which are often accompanied by depression, eating disorders and/or drug abuse.

The types of anxiety disorders falling within the category of anxiety disorders include panic disorder (panic attacks without symptoms accompanied by sudden phobias and physical symptoms including chest pain, palpitations, shortness of breath, dizziness, abdominal discomfort, unrealistic sensations and panic death) as well as social phobias and specific phobias (the former involving paralytic, irrational self-awareness of social circumstances and the latter involving specific phobias such as unprovoked fear of flight, blood or altitude).

Another type of anxiety disorder is post-traumatic stress disorder, which may be caused by a terrible experience in which severe physical injury occurs or is threatened. Posttraumatic stress disorder can develop from rape, child abuse, survivors of war or natural disasters. Common symptoms include hallucinations reappearance (the person in the hallucinations experiences afraid of experience), nightmares, depression, and feeling angry or irritable.

Obsessive compulsive disorder is another type of anxiety disorder. This is a condition in which people experience persistent unwanted thoughts (obsessions) and/or actions (obsessions) that they cannot control. In general, obsessive ideas are related to staining, suspicion (e.g., worrying about not turning off the household appliance), and confusion of sexual or religious ideas. The forced behavior includes cleaning, inspection, scheduling and counting.

Generalized anxiety disorder is another type of anxiety disorder in which a person repeatedly, exaggeratedly worrys about daily life events and activities. The disorder often persists for months, during which time the patient has more days of extreme concern than if it had not. The individual thinks of the worst case even though the other person says he or she has no reason to anticipate the situation. Physical symptoms may include nausea, thrillery, fatigue, muscle tension, and/or headache.

There are two main medical approaches to the treatment of anxiety disorders: (1) drug therapy and (2) cognitive-behavioral therapy (CBT). Combinations of the two types of treatment may also be effective. Because most anxiety disorders have at least some biological components, antidepressants and anxiolytics are often prescribed.

Inflammatory bowel disease

Inflammatory Bowel Disease (IBD) is a group of inflammatory disorders of the colon and small intestine. The main types of IBD are crohn's disease and ulcerative colitis. IBD can present with any of the following symptoms: abdominal pain, vomiting, diarrhea, blood in the stool (bright red blood in the stool), and weight loss. Diagnosis is usually by colonoscopy and biopsy of the pathological lesions.

While IBD may limit quality of life due to pain, vomiting, diarrhea, and other socially unacceptable symptoms, IBD itself is rarely fatal. Mortality due to complications such as toxic megacolon, intestinal perforation and surgical complications is also low. IBD patients do have an increased risk of colorectal cancer, but these patients are usually monitored regularly for this cancer, and therefore their colorectal cancer is usually detected earlier than in the average.

Treatment of IBD depends on the severity of the particular condition. IBD may require immunosuppression or a form of 5-aminosalicylic acid. In general, steroids are used to control disease flare-up. TNF inhibitors may also be useful in patients with Crohn's disease and patients with ulcerative colitis. Severe cases may require surgery, such as bowel resection, stricture plasty, or temporary or permanent colostomy or ileostomy.

The goal of treatment is to achieve relief, after which patients are often switched to less potent drugs with fewer potential side effects. Sometimes, an acute reproduction of the initial symptoms may occur. Depending on the situation, this may disappear on its own or require medication. The time between such reproductions may be anywhere from weeks to years, with large differences between patients.

Irritable bowel syndrome

Irritable Bowel Syndrome (IBS) is a condition most often characterized by spasticity, abdominal pain, bloating, constipation, and/or diarrhea. IBS can cause a great deal of discomfort and distress, but it does not permanently damage the gut and does not lead to serious diseases such as cancer. Many people can control their symptoms through diet, stress management and prescription medications. For some people, however, IBS may be disabling. They may be unable to work, participate in social activities, or even travel for short distances.

Up to 20% of adults have IBS symptoms, making it one of the most common conditions diagnosed by physicians. Its incidence in women is usually greater than in men, beginning before the age of 35 in about 50% of patients. Sometimes, people find their symptoms to resolve for months and then recover, while others report that symptoms are worsening over time.

There is no specific diagnostic test for IBS, but diagnostic tests can be performed to eliminate other problems. These tests may include stool sample tests, blood tests, and x-rays. Generally, the physician will perform a sigmoidoscopy or colonoscopy. Physicians can diagnose IBS based on the patient's symptoms, including the frequency of abdominal pain or discomfort over the past year, when pain associated with bowel function begins and ends, and how the frequency of bowel movements (bowel frequency) and stool consistency change.

Unfortunately, many people suffer from IBS for a long time before medical treatment is sought. Up to 70% of people with IBS do not receive medical treatment for their symptoms. Pharmacological treatment is an important part of alleviating the symptoms of IBS. Such medications include fiber supplements or laxatives for constipation, drugs to reduce diarrhea, and antispasmodics to control colonic muscle spasm and reduce abdominal pain. In addition, antidepressants may alleviate some IBS symptoms.

Inflammatory airway diseases

Inflammatory airway diseases include asthma and Chronic Obstructive Pulmonary Disease (COPD). Asthma is a chronic inflammation of the lungs in which the airways (bronchi) in the lungs become reversibly narrowed. Asthma affects 7% of the population, with 3 billion people worldwide. During an asthma attack, smooth muscle cells in the bronchi contract and the airways become inflamed and swollen. This can lead to breathing difficulties.

Asthma causes approximately 4,000 deaths annually in the united states. Seizures can be prevented by avoiding triggers and by medication. Drugs such as inhaled β 2 agonists are often used for acute episodes. In more severe cases, the drug is used for long-term prophylaxis, starting with inhaled corticosteroids, and then (if needed) long-acting β 2 agonists. Leukotriene antagonists may also be used in place of corticosteroids. Monoclonal antibodies, such as mepolizumab (mepolizumab) and omalizumab (omalizumab), are sometimes effective.

COPD includes a small number of lung diseases such as chronic bronchitis and emphysema. Many people with COPD have both of these diseases. Symptoms of COPD include shortness of breath, increased mucus in the lungs, and coughing. The main COPD treatments are: smoking cessation, pharmaceutical treatments such as bronchodilators and corticosteroids, and lung rehabilitation.

Urinary incontinence

Urinary incontinence is the inability to control the release of urine from the bladder. Some people develop occasional small urine leaks, while others often wet their clothing. Types of urinary incontinence include stress incontinence, urge incontinence and overflow incontinence. Treatment of urinary incontinence depends on the type of incontinence, the severity of the problem, and the underlying cause. Treatment may include, for example, behavioral techniques, physical therapy and/or drug therapy such as anticholinergics, topical estrogens and imipramine.

The limited efficacy, often unacceptable side effects, and physiological factors that may induce or otherwise affect the disorders and disease processes discussed above, make it necessary to continue to search for new compounds with new pharmacological effects to address these disorders and diseases.

Disclosure of Invention

The present invention relates to a compound having the structure:

wherein:

(i) a and B are independently-OH or-SH,

(ii) v and W are independently oxygen or sulfur, and at least one of V and W is oxygen,

（iii）R₁is- (CH)₂)_pCH₃Or is-H, and

(iv) p is an integer from 0 to 3, and:

(A) x is- (CH)₂)_m-，

(B) Y is-H, and Y is-H,

(C) z is- (CH)₂)_n-，

(D) m and n are integers which are defined as,

(E) m =1 to 5, and m is a linear chain,

(F) n =4 to 14, and n is a linear variable,

(G) for all m and n, 6. ltoreq. m + n. ltoreq.14, and

(H) wherein, optionally, there are up to two carbon-carbon double bonds, each double bond formed between adjacent methylene groups of formula (1), wherein if two of said double bonds are present, each carbon atom thereof is bonded to at least one hydrogen;

or (I) X is

(J) Y is absent, C_AAnd C_BTogether form a double bond, which is,

(K) z is- (CH)₂)_r-，

(L) q and r are integers,

(M) q =0 to 4,

(N) r =1 to 13,

(O) for all q and r, 5. ltoreq. q + r. ltoreq.13, and

(P) wherein, optionally, a second double bond is formed between adjacent methylene groups of formula (1) wherein each carbon atom thereof is bonded to at least one hydrogen;

or (Q) X is- (CH)₂)_t-，

(R) Z is

(S) Y is absent, C_AAnd C_cTogether form a double bond, which is,

（T）R₁is- (CH)₂)_vCH₃Or is a compound of formula (I) or a compound of formula (II),

(U) t and U are integers,

(V) t =1 to 5,

(W) u =0 to 12,

(X) for all t and u, 5. ltoreq. t + u. ltoreq.13,

(Y) wherein, optionally, a second double bond is formed between adjacent methylene groups of formula (1) wherein each carbon atom thereof is bonded to at least one hydrogen.

In one aspect of the invention, A and B are both-OH.

V and W may both be hydrogen.

Preferably, R₁Is- (CH)₂)_pCH₃. The value of p can be 0 to 2, more preferably p is 0 or 1, most preferably p is 0.

n may have a value of 2 to 12 and 7. ltoreq. m + n. ltoreq.13, or n may have a value of 3 to 11 and 8. ltoreq. m + n. ltoreq.12, or n may have a value of 4 to 10 and 9. ltoreq. m + n. ltoreq.11, more preferably n is 5 to 9 and m + n = 10. The value of m may be 2 to 4, but is preferably 3.

The value of r can be from 2 to 12, with 6. ltoreq. q + r. ltoreq.12; more preferably r is 3 to 11 and 7. ltoreq. q + r. ltoreq.11; more preferably r is 4 to 10 and 8. ltoreq. q + r. ltoreq.10; most preferably r is 5 to 9 and q + r is 9.

q may have a value of 1 to 3, preferably q is 2.

u can have a value of 1 to 11 and 6. ltoreq. t + u. ltoreq.12; more preferably u is from 2 to 10 and 7. ltoreq. t + u. ltoreq.11; more preferably u is from 3 to 9 and 8. ltoreq. t + u. ltoreq.10; most preferably u is 4 to 8 and t + u is 9.

the value of t may be 2 to 4, preferably 3.

If one or both of the two carbon-carbon double bonds of the preceding paragraph (H) are present in the compound, each of these double bonds may form between the methylene groups of Z. The methylene group being- (CH)₂)-. Preferably, if such a bond is present, only one of them is present.

If a second double bond as described in paragraph (P) is present, this bond is preferably formed between the methylene groups of Z.

Most preferably, the double bonds of both paragraphs (H) and (P) are absent.

Preferred compounds have formula (I) which is a mixture of all 4 stereoisomers. When referring to stereoisomers herein, the inventors speak of stereoisomers arising from the presence of two chiral centers, such as those stereoisomers found in the compounds of formula (I) and discussed further below.

Formula (I)

Preferred compounds are substantially stereochemically pure compounds of formula (I) of the following structure:

another preferred compound is a substantially stereochemically pure compound of formula (I) of the following structure:

another preferred compound is a substantially stereochemically pure compound of formula (I) of the following structure:

another preferred compound is a substantially stereochemically pure compound of formula (I) of the following structure:

throughout this specification, formula (I) is commonly referred to as 6-methylmyristolein, more commonly referred to as 6-MMAM. When a compound of the invention is referred to as having formula (I) without further recitation, this means that the compound is a mixture of the 4 stereoisomers described above.

The invention includes compounds of formula (I) wherein the chiral carbon atom of the glycerol moiety is a mixture of R and S stereochemical configurations, and the C-6 carbon of the myristic acid moiety is in the R configuration. Also included are compounds of formula (I) wherein the chiral carbon atoms of the glycerol moiety are a mixture of two stereochemical configurations R and S, while the C-6 carbon of the myristic acid moiety has the S configuration. Furthermore, the invention includes compounds of formula (I) wherein the chiral carbon atom of the glycerol moiety has the S configuration and the C-6 carbon of the myristic acid moiety is a mixture of R and S. Also included are compounds of formula (I) wherein the chiral carbon atom of the glycerol moiety has the R configuration and the C-6 carbon of the myristic acid moiety is a mixture of R and S.

The invention includes pharmaceutical compositions comprising any of the above compounds.

The pharmaceutical composition may be adapted for oral, parenteral, topical, rectal, vaginal, oral inhalation or nasal delivery.

The present invention includes any of the foregoing compounds in various forms. Specific dosage forms include solutions, suspensions, syrups, tablets, capsules, microgranules, ointments, creams or lozenges, or capsules, with the preferred dosage form being a tablet.

The invention includes a method for treating neurokinin 2 (NK)₂) A disorder or disease associated with receptor activity. The method comprises the following steps: administering a therapeutically effective amount of any of the foregoing compounds. When reference is made herein to a compound of the invention, whether explicitly stated or not, it is to be understood as including pharmaceutically acceptable salts.

Treated according to the method of the invention and the NK₂The disorder or disease associated with receptor activity may be a depressive mood disorder, an anxiety disorder, irritable bowel syndrome, inflammatory bowel disease, inflammatory airway disease or urinary incontinence. And said NK₂The disorder or disease associated with receptor activity may be in particular a depressive mood disorder, or it may be a major depressive disorder.

The subject or patient may be treated by psychotherapy or not by psychotherapy while being treated by the method of the invention.

Of course, the compounds of the present invention may be included in a pharmaceutical formulation which also includes a pharmaceutically acceptable carrier.

Administration of a compound described herein may be accompanied by a therapeutically effective amount of another therapeutic agent.

Generally, the subject treated with the present invention is a human patient.

Another method of the invention is for treating a disorder or syndrome associated with depressive mood disorders. The method comprises the following steps: administering to a subject in need thereof a therapeutically effective amount of a compound of the invention. The disorder or syndrome may be a brain or nervous system disorder, an anxiety disorder, a sexual dysfunction, drug abuse, an eating disorder, or a hormonal disorder.

In another aspect, the invention relates to a method of treating a disorder or condition treatable by an antidepressant. The method comprises administering to a subject in need thereof a therapeutically effective amount of a compound of the invention. The disorder or condition treatable by an antidepressant may be hot flashes associated with menopause, pain or smoking cessation.

Another method of the invention for regulated NK₂(ii) the activity of a receptor comprising the NK₂The receptor is contacted with an effective amount of a compound of the invention. The method may be an in vivo method or an in vitro method.

The invention includes the use of a compound as described above, or a pharmaceutically acceptable salt thereof, in combination with the various methods of the invention for the treatment of a disorder or disease, etc., as described above.

An innovative use of the compounds of the invention therefore also lies in the preparation of medicaments for the treatment of such disorders or diseases and the like.

Drawings

One skilled in the relevant 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.

Figure 1 shows an HPLC chromatogram of fertilized egg isolate according to an embodiment of the present invention.

FIG.2 shows the results of an analysis of fertilized egg isolates according to an embodiment of the present invention.

FIG.3 shows fertilized egg isolate sample #20 supernatant isolates (μ g/mL) versus neurokinin A (NKA) and human NK at various concentrations₂Graph of the effect of receptor binding (measured as percent specific binding), and IC of NKA and sample #20 supernatant isolate₅₀And K_i。

FIG.4 shows multiple fractions of formulation A and control samples (eluted from HPLC) for human NK₂Bar graph of the binding activity of the receptor. Binding activity was measured as percent inhibition of ligand NKA binding.

Figure 5 shows an HPLC-UV chromatogram of the 171 fraction of formulation a. The UV detector was set at 210 nm. The x-axis is in units of time (minutes) and the y-axis is in units of Absorbance (AU).

Figure 6 shows the HPLC-UV chromatogram of the 185 fraction of formulation a. The UV detector was set at 210 nm. The x-axis is in units of time (minutes) and the y-axis is in units of Absorbance (AU).

Figure 7 shows the HPLC-UV chromatogram of the 171 fraction of formulation a. The UV detector was set at 190 nm. The x-axis is in units of time (minutes) and the y-axis is in units of Absorbance (AU).

Figure 8 shows the HPLC-UV chromatogram of the 185 fraction of formulation a. The UV detector was set at 190 nm. The x-axis is in units of time (minutes) and the y-axis is in units of Absorbance (AU).

FIG.9 shows various concentrations of 2, 3-dihydroxypropyl 6-methyl-myristate to neurokinin A (NKA) and human NK₂Graph of the effect of receptor binding (measured as percent specific binding), and the IC of NKA and 2, 3-dihydroxypropyl 6-methyl-myristate₅₀And K_i。

FIG.10 shows various concentrations of 2, 3-dihydroxypropyl myristate vs. neurokinin A (NKA) and human NK₂Graph of the effect of receptor binding (measured as percent specific binding), and the IC of NKA and 2, 3-dihydroxypropyl myristate₅₀And K_i。

FIG.11 shows Ca on cells/function²⁺bAla in agonist assay⁸-NKA (4-10) (control), Compound #2 (2, 3-dihydroxypropyl myristate) and Compound #3 (2, 3-dihydroxypropyl 6-methyl-myristate) to human NK₂Graph of the effect of receptors. The x-axis represents the log of the compound (M) and the y-axis represents the% maximal Response (RFU). Error bars are the range of parallel data.

FIG.12 showsIn the cell/function Ca²⁺bAla in agonist assay⁸-NKA (4-10) (control), GR159897 (control), Compound #2 (2, 3-dihydroxypropyl myristate) and Compound #3 (2, 3-dihydroxypropyl 6-methyl-myristate) to human NK₂Graph of the effect of receptors. The x-axis represents the log of the compound (M) and the y-axis represents the% maximal Response (RFU). Error bars are the range of parallel data.

Detailed Description

According to the invention, the compounds of the invention are described, as well as the treatment and Neurokinin (NK)₂) A disorder or disease associated with receptor activity.

The term "pharmaceutically acceptable salt" as used herein is a salt formed from an acidic group or a basic group of a compound having the structural formula of the present invention. Exemplary salts are known to those skilled in the art and include, but are not limited to, hydrochloride, sulfate, citrate, acetate, oxalate, chloride, bromide, iodide, nitrate, bisulfate, phosphate, acid phosphate, isonicotinate, lactate, salicylate, acid citrate, tartrate, oleate, tannate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisate (gentisate), fumarate, gluconate, glucuronate (glucaronate), saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, and pamoate (i.e., 1' -methylene-bis (2-hydroxy-3-naphthoate).

According to the present invention, the chemical structures described herein (including the compounds of the present invention) encompass all enantiomers and stereoisomers of the corresponding compounds, that is, stereomerically pure forms (e.g., geometrically pure, enantiomerically pure or diastereomerically pure forms) as well as mixtures of enantiomers, diastereomers and geometric isomers, and the like, are also compounds of the present invention. Methods for separating one enantiomer from another are known to those skilled in the art. In some cases, one enantiomer, diastereomer or geometric isomer will have superior activity or improved toxicity or kinetic properties compared to the others. In these cases, such enantiomers, diastereomers and geometric isomers in the compounds of the invention are preferred.

The compounds of the present invention, including any enantiomer of the compound, may be substantially pure. A compound is "substantially pure" when it is isolated from a component with which it is naturally found. Thus, for example, when a compound of formula (I) isolated from a fertilized egg isolate is separated from the other components of the fertilized egg isolate, it will generally be substantially pure. In general, a compound is substantially pure when it is present at least 60, 70, 75, 80, 85, 90, 95, or 99 weight percent of the total material in the sample. Substantially pure compounds can be obtained, for example, by extraction from natural sources (e.g., fertilized egg isolates) or by chemical synthesis. Purity can be measured using any suitable method, such as column chromatography, gel electrophoresis, High Pressure Liquid Chromatography (HPLC), and the like.

The compounds of the invention are useful in therapy and NK₂A disorder or disease associated with receptor activity. The compounds of the present invention may be administered to a subject in need thereof in a therapeutically effective amount.

The term "treatment" refers to ameliorating a disorder or disease in a patient to whom a compound of the invention is administered. The term "treatable" refers to a disorder, disease or condition that improves a patient to whom a compound of the invention is administered. These terms include, for example, ameliorating a disorder, disease, or condition by obtaining a beneficial result, such amelioration can be determined using standard tests known in the art. The term also includes preventing the occurrence or recurrence of the disorder or disease, e.g., prophylactic or maintenance therapy.

Terms used herein“NK₂Receptor-related disorders or diseases "refers to inappropriate NK₂Receptor activity (e.g., above or below normal NK)₂Receptor activity) associated disorder or disease. More normal NK₂The high receptor activity is probably due to the normal number of NK in the subject₂Increased activity of the receptor, or possibly due to NK₂NK in subjects with receptor-related disorders or diseases₂The number of receptors is higher than normal. More normal NK₂The low receptor activity may be due to a normal number of NK in the subject₂Reduced receptor activity, or may be due to NK₂NK in subjects with receptor-related disorders or diseases₂The number of receptors is less than normal. NK₂Receptor-related disorders or diseases include, for example, major depressive disorder, anxiety disorder, irritable bowel syndrome, inflammatory bowel disease, inflammatory airway disease, and urinary incontinence. NK₂The receptor-related disorder or disease may include at least in part NK₂A receptor-mediated disorder or disease.

An "effective amount" is the amount of a compound that, when administered to a subject suffering from NK₂The amount of a compound that achieves a beneficial result in a subject suffering from a disorder or disease associated with receptor activity, or the amount of a compound that has a desired activity in vivo or in vitro. For and NK₂In the case of a disorder or disease associated with receptor activity, a beneficial outcome includes a reduction in the extent or severity of symptoms associated with the disorder or condition, and/or an increase in the quality of life of the subject as compared to when not treated. For example, for a subject with major depressive disorder, a "favorable outcome" includes the Hamilton Depression Rating Scale (the Hamilton Depression Rating Scale), Hamilton Anxiety Rating Scale (the Hamilton Anxiety Rating Scale), montgomery-emberg Depression Rating Scale (the) of the subject as compared to the score of a subject not treated with a compound of the inventionSuppression Rating Scale), Beck Depression scoreA reduction in the rating Scale (the Beck prediction Inventory), the Arizona serial Experience Scale (the Arizona serial Experience Scale) or the General Health Questionnaire rating (profile 36), each of which is known to those skilled in the art and will be described in further detail herein.

For a subject with an anxiety disorder, a "favorable outcome" includes a decrease in the hamilton anxiety rating scale score of the subject as compared to a subject not treated with a compound of the invention; decreased mood or decreased frequency of mood, both distress and fright; a reduction in the number and/or duration of panic attacks; a reduction in escape from social circumstances; reduced fear associated with specific phobias; reduced occurrence or duration of hallucinations, nightmares, depression and irritation or irritability associated with post-traumatic stress disorder; and a reduction in the occurrence of obsessions and/or compulsions.

For patients with inflammatory bowel disease, "favorable results" include a reduction in abdominal pain, vomiting, diarrhea, hematochezia, and/or weight loss as compared to subjects not treated with a compound of the invention.

For patients with irritable bowel syndrome, "favorable results" include a reduction in spasticity, abdominal pain, bloating, constipation, and/or diarrhea as compared to a subject not treated with a compound of the invention.

For a patient with an inflammatory airway disease, a "favorable outcome" includes a reduction in the frequency and/or duration of shortness of breath, reduction of mucus in the lungs, and/or cough episodes as compared to a subject not treated with a compound of the invention.

For patients suffering from urinary incontinence, "favorable results" include reduced urine leakage and/or wetting of clothing, as compared to subjects not treated with the compounds of the present invention.

The precise amount of the compound administered to a subject will depend upon the type and severity of the disorder or disease and upon the characteristics of the subject, such as general health, age, sex, body weight and tolerance to drugs. One skilled in the art will be able to determine the appropriate dosage based on these and other factors.

The terms "subject," "patient," and "animal" as used herein are used interchangeably and include, but are not limited to, cattle, monkeys, horses, sheep, pigs, chickens, turkeys, quail, cats, dogs, mice, rats, rabbits, guinea pigs, and humans. In one embodiment, the subject, patient, or animal is a mammal. In another embodiment, the preferred subject, patient or animal is a human.

Methods of isolating the compounds of the invention:

the compound having the structure represented by formula (I) can be isolated from fertilized eggs as described below.

Fertilized egg isolate-preparation

In preparing fertilized egg isolates from which a compound of formula (I) may be isolated, at least one fertilized egg is incubated for any of about 3 to about 15 days, more preferably about 3 to about 5 days, or more preferably about 6 to about 12 days, and even 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 to allow angiogenesis to begin and/or to allow the embryo to mature until the embryo is visible 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 a blood vessel connected to an embryo can be removed is suitable. The eggs are preferably avian eggs, and may be obtained from any avian species that has been subjected to egg reproduction, such as chickens, geese, ducks, and the like. Chicken eggs are preferred for reasons including their availability and ability to be mass produced. Incubation can be performed in any environment, as long as the eggs are kept at one temperature for a long time to mature the embryo. Suitable incubation temperatures range from about 20 ℃ to about 60 ℃, more preferably from about 25 ℃ to about 55 ℃, and more preferably from about 35 ℃ to about 45 ℃. Once the eggs have been incubated for a period of time, they may optionally be treated to reduce the external microbiota or sterilise them by any suitable method 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 by 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 ℃.

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 for filtration. The mesh size may be 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 ruptured egg shells to filter on the screen for a period of time such that substantially no more liquid drips 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 solids 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.

The white albumen portion and/or embryo may be substantially separated from other egg contents. The white egg white portion may be substantially separated from the other contents by any suitable method (e.g., pouring out the white egg white portion) or by suction. The embryo may be substantially separated from the white albumen portion manually or by other suitable methods as determined by the skilled person. One skilled in the art will recognize that embryos may be substantially separated from white albumen portions 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 yolk sac of the embryo, which forms part of the other contents, may be manually peeled off.

Once the embryo is substantially separated from the white albumen 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 have been 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 the intact fertilized egg may be broken, the egg shell removed, and the entire shelled egg frozen and freeze-dried according to any of the methods described above and below to produce a fertilized egg isolate. Alternatively, more than one intact zygote may be broken, the shell removed, the entire shelled zygote combined and mixed into a slurry, then frozen and freeze-dried 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 being removed 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.

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 using, for example, a hand-held mixer or other suitable tool. The slurry was then frozen and freeze dried as described above. Preferably, the final temperature at which lyophilization is carried out is 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 ℃, and 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 final 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 freeze-drying process is typically 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.

The freeze-dried contents, freeze-dried embryos, or freeze-dried slurry may then be dispersed and/or comminuted as desired to form a substantially homogeneous powder. The contents, separately or in smaller groups, that are freeze-dried may be combined together before or after the comminuting step to form a substantially uniform powder. The comminution may 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. A suitable sterilization process is one that should not adversely affect certain freeze-dried components.

Preservatives to control microbial growth may be incorporated into the powder or concentrate according to any of the methods described herein prior to storage. Instead of the preservative, it may also be added at another stage of the preparation (including prior to the freeze-drying or concentration step) instead of to the powder or concentrate; or the preservative may also be added at another stage of the manufacturing process (including prior to the freeze-drying or concentration step) and may be added 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 at a temperature of 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.

Slurries can also 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. The slurry may be freeze-dried as described above or used partially or wholly in the extraction step as described below.

The slurry may also be mixed with the aqueous solution for a period of time. The aqueous solution may comprise water, an aqueous buffer, 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 contents such that any substantially hydrophilic molecules in the solution are dissolved in the aqueous solution. The volume of 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, the aqueous solution may be substantially clarified by substantially removing any solid portion of the mixture by a suitable method, such as centrifugation or filtration. The clarified aqueous portion can then be frozen and freeze-dried according to the methods described herein to produce a powder, which can optionally be sterilized.

The slurry produced by any of the above methods may be mixed with a substantially hydrophobic solvent. The substantially hydrophobic solvent is preferably frozen. 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 a suitable method, such as centrifugation or filtration. The solvent portion will comprise predominantly a hydrophobic solvent portion and may also comprise an aqueous portion. 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, allowing the top ether-containing layer to be poured out. The aqueous portion may be extracted multiple times, such as 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 the 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.

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 produced by the filtration step prior to further clarification^TM. Some of the resulting filtrate may be frozen in a suitable container for freeze-drying. In addition, some of the resulting filtrate may be mixed with the above-mentioned hydrophobic solvent so that an aqueous layer and a hydrophobic layer are formed. The layers can be separated, concentrated, and stored as described herein.

Fertilized egg isolates prepared by the various methods described herein may be concentrated by repeated aqueous and/or hydrophobic solvent extractions.

Compounds having the structure depicted in formula (I) can be isolated from the fertilized egg isolate using, for example, standard column chromatography techniques. For example, a slurry of the fertilized egg isolate may be prepared and freeze-dried as described above. The freeze-dried product may then be comminuted in a mill and, if desired, mixed with one or more preservatives, such as sodium benzoate (e.g. 0.5% w/w) and/or potassium sorbate (e.g. 0.2% w/w). The finally prepared powder can then be loaded onto a High Pressure Liquid Chromatography (HPLC) column and eluted with a suitable solvent, such as various concentrations of methanol or acetonitrile or mixtures of solvents. The compound fractions of the eluate are collected and dehydrated or, if desired, subjected to another round of column chromatography using, for example, a different column, a different solvent, or a different solvent concentration.

The purity of the desired fraction can be monitored using, for example, HPLC or other methods known to those skilled in the art, and (if desired) the fraction can be further purified using techniques known to those skilled in the art.

Once the fraction or combination of fractions is sufficiently pure, the structure of the active compound and its biological activity can be determined using methods known to those skilled in the art. For example, the biological activity of the active compounds can be determined using NK₂Receptor binding assays and/or receptor activity assays known to those skilled in the art.

Synthesis of Compounds of the invention

A stereoisomer mixture of the preferred compound of the invention, 6-methylmyristoleic acid monoglyceride, can be synthesized according to scheme a.

Thus, racemic 2-methyldecanal [19009-56-4] was reacted with triphenylbutanoic acid phosphonium bromide [17857-14-6] to give 6-methyl-4-ene-tetradecanoic acid after purification. Similar acid chlorides were prepared with thionyl chloride and treated directly with racemic isopropylidene glycerol, followed by hydrogenation. Isopropylidene was removed with HCl to give 6-methylmyristic acid 1-glyceride as a mixture of stereoisomers.

Scheme A

1. The dioxolane of step III can be obtained from Sigma-Aldrich, st.louis, Missouri, u.s.a., or can be prepared according to the route shown in scheme B.

2. For the sake of brevity: the 6-methylmyristoleic acid monoglyceride is referred to herein as 6-MMAM. 6-MMAM is also known as 6-methylmyristate 1-glyceride and 6-methylglycidyl myristate.

Route B

6-MMAM has two chiral centers and thus four stereoisomers exist:

the synthetic route shown in scheme a can yield a mixture of all 4 stereoisomers, but can be modified to yield the stereoisomer of 6-MMAM.

Thus, step III of scheme a can be modified by using the indicated stereoisomer of 2, 2-dimethyl-1, 3-dioxolane (R isomer or S isomer at C ″) or other suitable 1, 3-dioxolane. U.S. Pat. No.6,143,908 to Hinoue et al describes a method of preparing 1, 3-dioxolane-4-methanol compounds of scheme C.

Scheme C

Hinoue et al show that preferred examples of compound (1) of scheme C are 3-chloro-1, 2-propanediol and 3-bromo-1, 2-propanediol, R of said compound¹And R²May be the same or different and may be hydrogen, C₁-C₄Alkyl or phenyl. The dioxolane introduced in step III of scheme A corresponds to R in scheme C¹And R²All methyl groups. In other words, the use of acetone in step A of scheme C will result in the formation of the dioxolane shown in scheme A. Hinoue et al demonstrated the preparation of (S) -2, 2-dimethyl-1, 3-dioxolane-4-methanol using (R) -3-chloro-1, 2-propanediol as the starting compound in scheme C.

(R) -3-chloro-1, 2-propanediol (CAS No. 57090-45-6) and (S) -3-chloro-1, 2-propanediol (CAS No. 60827-45-4) are available from TCI America,9211N.Harborgate Street, Portland, OR 97203, U.S.A., and thus they can be used to produce (S) -2, 2-dimethyl-1, 3-dioxolane-4-methanol and (R) -2, 2-dimethyl-1, 3-dioxolane-4-methanol.

(R) -4-chloromethyl-2, 2-dimethyl-1, 3-dioxolane (CAS No. 57044-27-3) and (S) -4-chloromethyl-2, 2-dimethyl-1, 3-dioxolane (CAS No. 60456-22-6) are available from Ivy Fine Chemicals Corporation of 1879 Old Cuthbert Road, Suite 23, Cherry Hill, N.J. 08034, USA. Either of them can be introduced into step B of scheme C (Hinoue et al method) because R¹=R²Methyl so that step a of scheme C is not required.

Examples of hydrolysis of 1, 3-dioxolane under mild conditions in step III of scheme A are described, for example, in J.Sun, Y.Dong, L.Cao, X.Wang, S.Wang, Y.Hu, J.org.Chem.,2004,69: 8932-.

Second handedness center (C) in 6-MMAM⁺) In the first step of scheme A via 2-methyldecanal.

As shown in scheme D, the synthesis of scheme a can be modified to obtain other compounds of formula (1).

Scheme D

The materials may be selected according to the general formula shown in scheme D to give compounds wherein A and B are both-OH, V and W are both oxygen, and X is- (CH)₂)₄- (m = 3), Y is-H and Z is- (CH)₂)₆Inventive compounds of formula (1) of- (n = 7). The reaction conditions for each step can be varied by the skilled artisan to suit the particular material selected.

The preferred compound, 6-methylmyristic acid monoglyceride of the present invention, may also be synthesized according to scheme E, for example.

Route E

1. The dioxolane of step VIII can be prepared according to the route shown in scheme B.

2.R.O.Adlof，W.E.Neff，E.A.Emken,and E.H.Pryde,Journal of the American Oil Chemists'Society,1977,54(10):414-416。

The synthetic route shown in scheme E may yield a mixture of all 4 stereoisomers, but may be modified to yield each of the 4 stereoisomers.

Thus, step VIII of scheme E can be modified as described in step III and scheme C of scheme A.

Second handedness center (C) in 6-MMAM⁺) Can be introduced by reducing the double bond in step X in scheme E. Compound (6) (product of step V of scheme E) can be prepared under conditions wherein longer alkyl chains are formed trans to each other as exemplified in the Witting reaction shown. Asymmetric hydrogenation of the C = C bond will result in the formation of the R configuration or the S configuration at C-6 of 6-MMAM. Asymmetric hydrogenation of C = C bonds is well known. See, for example, de Paule et al, U.S. Pat. No.6,878,665.

Scheme F

The materials may be selected according to the general formula shown in scheme F to give compounds wherein A and B are both-OH, V and W are both oxygen, R₁Is- (CH)₂)_PCH₃or-H, p is an integer of 0-3, X is- (CH)₂)_m-, Y is-H, Z is- (CH)₂)_nAnd m and n are integers (where m = 1-5 and n = 4-14). The reaction conditions for each step can be varied by the skilled artisan to suit the particular material selected.

A stereochemically pure compound of the present invention is one wherein at least 90% of said compound has the desired stereochemistry (e.g., at C)⁺Is at R and is at C^*Is at S or at C⁺At R, S and at C^*Where R, etc.). More preferably, the compound is 92% stereochemically pure, still more preferably 94% stereochemically pure, still more preferably 96% stereochemically pure, still more preferably 98% stereochemically pure, most preferably greater than 99% stereochemically pure. A substantially stereochemically pure compound is at least 96% of the desired optical stereoisomer(s).

A method of preparing a compound of formula a6 or a7 is shown in scheme G. The process comprises reacting compounds a1 and a2 to form alkenyl compound A3. The acyl halide of A3 is then formed, which reacts with dioxolane a4 to form a5, a5 can then be hydrolyzed to form a6 or a 5C = C double bond reduction, followed by dioxolane hydrolysis to form compound a 7.

Scheme G

Wherein: a is an integer of 1 to 3; b is an integer of 1 to 11; a + b is more than or equal to 4 and less than or equal to 12 and R₁Is- (CH)₂)_PCH₃(ii) a And p is an integer from 0 to 3. Preferably, p is 0. R₂And R₃Which may be the same or different, may be any suitable group suitable for the reaction. Specific groups include those disclosed by Hinoue: hydrogen, C₁-C₄Alkyl or phenyl.

The R or S stereoisomer of dioxolane A4 can be used to obtain A6 or A7 which are, if desired, stereochemically pure at C-2 of the glycerol moiety of A6 or A7.

Functional analogs of compounds of formula (I):

functional analogs of the compounds of formula (I) can be prepared using methods known to those skilled in the art. For example, a compound of formula (I) isolated and identified or synthesized according to the methods described above can be chemically modified, directionally or randomly, e.g., to replace hydrogen with halogen, to replace one alkyl with a different alkyl, to replace alkoxy with alkyl, to replace alkyl with alkoxy, acylation, alkylation, esterification, amidation, etc., to produce structural analogs of the compound, which can be tested for biological activity (e.g., with NK) using the methods described herein or other methods known to those skilled in the art (e.g., with the methods described herein)₂Binding of the receptor, or receptor activity resulting in changes in intracellular calcium concentration).

Another way to obtain functional analogues of the compounds of formula (I) is by rational design. This can be achieved by structural information and computer modeling. Prediction of target molecule-compound interactions when small changes are made in one or both of the target molecule and compound can be made using molecular modeling software and computationally intensive computers. Examples of molecular modeling systems are CHARMm and QUANTA programs, Accelrys inc, San Diego, CA. CHARMm can perform energy minimization and molecular dynamics functions. QUANTA enables the construction, graphical modeling and analysis of molecular structures. QUANTA allows interactive construction, modification, visualization and analysis of the behavior of molecules with respect to each other. Other computer programs that can screen and graph chemical substances are known to those skilled in the art. Biological activity can also be tested for functional analogs by rational drug design using the methods described herein or other methods known to those skilled in the art (e.g., with NK)₂Binding of the receptor, or receptor activity resulting in changes in intracellular calcium concentration).

Therapeutic uses of the compounds of the invention:

as described in PCT publication No. WO2009/086634 (the entire teachings of which are incorporated herein by reference), fertilized egg isolates are useful in the treatment of patients suffering from mental health disorders, including depressive mood disorders, such as major depressive disorder, dysthymic disorder, depressive phase of bipolar disorder, depression caused by general medical conditions, such as depression associated with dementia or schizoaffective disorder, drug-induced depression and seasonal affective disorder, anxiety disorders, such as general anxiety disorder, social phobia, panic disorder, and sexual dysfunction.

As also described in PCT publication No. WO2009/086634, it has been determined that fertilized egg isolates described herein can antagonize the binding interaction of some ligands with their receptors. In particular, it has been found that the fertilized egg isolate has the ability to release the neurotransmitter neurokinin A (NKA) from its receptor neurokinin 2(NK₂) The ability to substitute at the receptor.

A number of diseases and disorders are also known, as are NK pairs₂The modulation of the receptor is relevant. Such diseases or conditions include depressive mood disorders, such as major depressive disorder (see, e.g., Dableh, Ahlstedt, Michale, Louis, Stein berg, Salom é, Holmes, Steinberg, Husum), anxiety disorders (see, e.g., Ahlstedt, Michale, Louis, Greibel, Steinberg, Stratton, Teixera, Walsh, Salom é, Holmes), irritable bowel syndrome and inflammatory bowel disease (see, e.g., Ahlstedt, Lecci, Evangelista, Tooulse), inflammatory airway diseases such as asthma or Chronic Obstructive Pulmonary Disease (COPD) (see, e.g., Bai, Pinto, Khawaja), and urinary incontinence (see, e.g., Ahldt, Rizzo). Furthermore, it has been demonstrated that NK₂Antagonists of receptors such as saredutant (SR 48964) are useful for promoting antidepressant-like effects (Salom, dableeh, Steinberg, Michale, Louis) and anxiolytic effects (Teixeira, Salom, Griebel, Michale, Louis) in animal models, and have been studied in humans. Regulation of NK₂Activation of the receptor (e.g. by inhibition of NK₂Binding of one or more endogenous ligands (e.g., NKA) to its receptor), and reduction or elimination of NK₂A disorder or disease associated with receptor activity.

A compound having the structure shown in formula (I) has been isolated from the fertilized egg isolate as described herein and has the ability to separate the neurotransmitter neurokinin A (NKA) from its human NK₂The ability to substitute at the receptor. Compounds of formula (I) have also been synthesized and found to convert the neurotransmitter neurokinin A (NKA) from human NK₂The receptor replaces and can alter downstream intracellular calcium levels. Thus, the compounds of formula (1), formula (I), and functional analogs and pharmaceutically acceptable salts thereof are useful in therapy and NK₂A disorder or disease associated with receptor activity.

Thus, another aspect of the present invention relates to a method of treatment and NK-treatment using compounds of formula (1), formula (I) and functional analogues and pharmaceutically acceptable salts thereof₂Activation of receptorA method of a sexually relevant disorder or disease, the method comprising the steps of: administering to a patient in need thereof a therapeutically effective amount of a compound of formula (1), formula (I) or a functional analogue or pharmaceutically acceptable salt thereof. The and NK₂The disorder or disease associated with receptor activity may be, for example, a depressive mood disorder, such as major depressive disorder, anxiety, inflammatory bowel disease, irritable bowel syndrome, inflammatory airway disease, or urinary incontinence.

It will be appreciated by those skilled in the art that the compounds of formula (1), formula (I) and functional analogues or pharmaceutically acceptable salts thereof may be useful in the treatment of depression-related disorders or conditions, such as disorders of the brain or nervous system, substance abuse, eating disorders and hormonal disorders, such as thyroid insufficiency, hypogonadism, menopause and the like. In addition, such compounds or analogs thereof or pharmaceutically acceptable salts thereof may be used to treat other conditions for which antidepressants have been shown to be effective, such as hot flashes associated with menopause, pain, and smoking cessation.

In therapy and NK₂In a method of treating a disorder, disease or condition associated with receptor activity, the patient may or may not be undergoing psychotherapeutic treatment concurrently with the treatment.

The compounds of the invention 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, sublingual and oral inhalation), nasal routes, topical routes (including buccal, sublingual and transdermal) or parenteral routes (including subcutaneous, intramuscular, intravenous or intradermal). 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.

The pharmaceutical composition comprises:

the present invention provides methods for treatment and NK₂Compositions of disorders or diseases related to receptor activitySuch as depressive mood disorders (e.g., major depressive disorder), anxiety disorders, irritable bowel syndrome, inflammatory bowel disease, inflammatory airway disease, or urinary incontinence. In one embodiment, the composition comprises one or more compounds of the present invention or a pharmaceutically acceptable salt thereof. In another embodiment, the compositions of the invention comprise one or more compounds of the invention, or pharmaceutically acceptable salts thereof, and one or more other prophylactic or therapeutic agents. In another embodiment, the composition comprises a compound of the invention, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier (diluent or excipient). In another embodiment, the composition is formulated such that it crosses the blood-brain barrier.

The compositions of the present invention may be pharmaceutical compositions or single unit dosage forms. The pharmaceutical compositions and dosage forms of the invention comprise one or more active ingredients in relative amounts and are formulated in the following manner: the given pharmaceutical composition or dosage form can be used for treatment and NK₂A disorder or disease associated with receptor activity. Preferred pharmaceutical compositions and dosage forms comprise a compound of formula (1), formula (I) or a functional analogue or pharmaceutically acceptable salt thereof, optionally in combination with one or more additional active agents.

The single unit dosage forms of the invention are suitable for oral, transmucosal (e.g., nasal, sublingual, vaginal, buccal, or rectal), parenteral (including subcutaneous, intravenous, bolus injection, intramuscular, or intraarterial) or transdermal administration to a patient. Examples of dosage forms include, but are not limited to: a tablet; caplets (caplets); capsules such as soft elastic gelatin capsules; a cachet; dragees (troch); a lozenge; a dispersant; suppositories; an ointment; cataplasms (cataplasms); a paste; powder preparation; a dressing; a cream; a plaster; a solution agent; patches (patch); aerosols (e.g., nasal sprays or inhalants); gelling agent; liquid dosage forms suitable for oral or mucosal administration to a patient include suspensions (aqueous or non-aqueous liquid suspensions, oil-in-water emulsions, or water-in-oil liquid emulsions), solutions, and elixirs.

The composition, shape and type of dosage forms of the present invention generally vary with their use. For example, a dosage form suitable for mucosal administration may contain a lower amount of active ingredient(s) than an oral dosage form for treating the same indication. This aspect of the invention is readily understood by those skilled in the art. See, e.g., Remington's Pharmaceutical Sciences (1990)18th ed., Mack Publishing, Easton, Pa.

Typical pharmaceutical compositions and dosage forms contain one or more excipients. Suitable excipients are well known to those skilled in the art of pharmaceutical and/or formulation chemistry, non-limiting examples of suitable excipients being given herein. Whether a particular excipient is properly incorporated into a pharmaceutical composition or dosage form depends on a variety of factors well known in the art, including, but not limited to, the manner in which the dosage form will be administered to a patient. For example, oral dosage forms (e.g., tablets) may contain excipients that are not suitable for use in parenteral dosage forms.

The invention also encompasses pharmaceutical compositions and dosage forms comprising one or more compounds that reduce the rate of decomposition of an active ingredient. Such compounds (referred to herein as "stabilizers") include, but are not limited to, antioxidants, such as ascorbic acid, pH buffers, and salt buffers.

Oral dosage forms:

pharmaceutical compositions of the present invention suitable for oral administration may be presented as discrete dosage forms such as, but not limited to, tablets (e.g., chewable tablets), caplets, capsules, and liquids (e.g., flavored syrups). Such dosage forms contain a predetermined amount of active ingredient and may be prepared by pharmaceutical methods well known to those skilled in the art. See generally Remington's Pharmaceutical Sciences (1990)18th ed., Mack Publishing, Easton, Pa.

The general oral dosage form of the present invention may be prepared by: the mixture of the active ingredient(s) and at least one excipient is combined according to conventional pharmaceutical compounding techniques. Excipients may take a wide variety of forms depending on the form of preparation desired for administration. For example, excipients suitable for use in oral liquid or aerosol dosage forms include, but are not limited to, water, glycols, oils, alcohols, flavoring agents, preservatives, and coloring agents. Examples of excipients suitable for use in solid oral dosage forms (e.g., powders, tablets, capsules, and caplets) include, but are not limited to, starches, sugars, microcrystalline cellulose, diluents, granulating agents, lubricants, binders, and disintegrating agents.

Because of their ease of administration, tablets and capsules represent the most advantageous oral dosage unit form, in which case solid excipients are used. If desired, the tablets may be coated by standard aqueous or non-aqueous techniques. Such dosage forms may be prepared by any pharmaceutical method. Generally, pharmaceutical compositions and dosage forms are prepared by: the active ingredient is uniformly and thoroughly mixed with the liquid carrier, finely divided solid carrier, or both, and then, if desired, the product is molded into the desired form.

For example, tablets may be prepared by compression or molding. Compressed tablets may be prepared by: the active ingredient in free-flowing form (e.g. powder or granules) is compressed in a suitable machine, optionally mixed with excipients. Molded tablets may be made by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent.

Examples of excipients that may be used in the oral dosage forms of the present invention include, but are not limited to, binders, fillers, disintegrants, and lubricants. Binders suitable for use in pharmaceutical compositions and dosage forms include, but are not limited to, corn starch, potato starch or other starches, gelatin, natural and synthetic gums such as acacia, sodium alginate, alginic acid, other alginates, powdered tragacanth, guar gum, cellulose and its derivatives (e.g., ethyl cellulose, cellulose acetate, carboxymethylcellulose calcium, carboxymethylcellulose sodium), polyvinylpyrrolidone, methyl cellulose, pregelatinized starch, hydroxypropylmethyl cellulose (e.g., nos. 2208, 2906, 2910), microcrystalline cellulose, and mixtures thereof.

Suitable forms of microcrystalline cellulose include, but are not limited to, those commercially available as AVICEL-PH-101, AVICEL-PH-103, AVICEL RC-581, AVICEL-PH-105 (available from FMC corporation, American Viscose Division, Avicel Sales, Marcus Hook, Pa.), and mixtures thereof. One specific binder is a mixture of microcrystalline cellulose and sodium carboxymethylcellulose commercially available as AVICELRC-581. Suitable anhydrous or low moisture excipients or additives include AVICEL-PH-103J and Starch 1500 LM.

Examples of fillers suitable for use in the pharmaceutical compositions and dosage forms disclosed herein include, but are not limited to, talc, calcium carbonate (e.g., granules or powder), microcrystalline cellulose, powdered cellulose, dextrates, kaolin, mannitol, silicic acid, sorbitol, starch, pregelatinized starch, and mixtures thereof. The binder or filler in the pharmaceutical composition of the present invention is generally present in about 50% to about 99% by weight of the pharmaceutical composition or dosage form.

Disintegrants are used in the compositions of the invention to provide tablets that disintegrate when exposed to an aqueous environment. Tablets containing too much disintegrant may disintegrate during storage, while tablets containing too little may not disintegrate at a desired rate or under desired conditions. Thus, a sufficient amount of disintegrant that is neither too much nor too little to adversely affect the release of the active ingredient should be used to form the solid oral dosage forms of the present invention. The amount of disintegrant used varies based on the type of formulation and is readily discernible to one of ordinary skill in the art. Typical pharmaceutical compositions comprise from about 0.5 to about 15% by weight of disintegrant, preferably from about 1 to about 5% by weight of disintegrant.

Disintegrants that may be used in pharmaceutical compositions and dosage forms of the invention include, but are not limited to, agar-agar, alginic acid, calcium carbonate, microcrystalline cellulose, croscarmellose sodium, crospovidone, polacrilin potassium, sodium starch glycolate, potato or tapioca starch, other starches, pregelatinized starches, other starches, clays, other algins, other celluloses, gums, and mixtures thereof.

Lubricants that may be used in the pharmaceutical compositions and dosage forms of the present invention include, but are not limited to, calcium stearate, magnesium stearate, mineral oil, light mineral oil, glycerol, sorbitol, mannitol, polyethylene glycol, other glycols, stearic acid, sodium lauryl sulfate, talc, hydrogenated vegetable oils (e.g., peanut oil, cottonseed oil, sunflower oil, sesame oil, olive oil, corn oil, or soybean oil), zinc stearate, ethyl oleate, ethyl laurate, agar, and mixtures thereof. Other lubricants include, for example, Syloid silica gel (AEROSIL 200, manufactured by Baltimore, Md., w.r.grace co.), a synthetic silica solidification aerosol (sold by Degussa co. of Piano, tex), CAB-O-SIL (a pyrogenic silica product sold by Cabot co. of Boston, mass), and mixtures thereof. Lubricants, if used, are generally used in amounts less than about 1% by weight of the pharmaceutical composition or dosage form into which they are incorporated.

Controlled release dosage forms:

the active ingredients of the present invention may be administered by controlled release means or by delivery devices well known to those of ordinary skill in the art. Examples include, but are not limited to, U.S. Pat. nos. 3,845,770; 3,916,899; 3,536,809, respectively; 3,598,123, respectively; and 4,008,719, 5,674,533, 5,059,595, 5,591,767, 5,120,548, 5,073,543, 5,639,476, 5,354,556, and 5,733,566, the entire teachings of each of which are incorporated herein by reference. Such dosage forms may be used to provide slow or controlled release of one or more active ingredients using, for example, hydroxypropylmethylcellulose, other polymer matrices, gels, permeable membranes, osmotic systems, multilayer coatings, microparticles, liposomes, microspheres, or combinations thereof to provide the desired release profile in various ratios. Suitable controlled release formulations known to those of ordinary skill in the art, including those described herein, can be readily selected for use with the compounds of the present invention. The present invention thus encompasses single unit dosage forms suitable for oral administration, such as, but not limited to, tablets, capsules, gelcaps, and caplets suitable for controlled release.

All controlled release drug products share the following common objectives: improved drug therapy compared to their uncontrolled counterparts. Ideally, the use of optimally designed controlled release formulations in medical treatment is characterized by the use of a minimum amount of drug substance to cure or control the condition in a minimum amount of time. Advantages of controlled release formulations include long-term drug activity, reduced dosing frequency, and increased patient compliance.

Most controlled release formulations are designed to initially release an amount of drug (active ingredient) that rapidly produces the desired therapeutic effect, and gradually and continuously release additional amounts of drug to maintain the level of therapeutic or prophylactic effect over an extended period of time. In order to maintain this constant level of drug in the body, the drug must be released from the dosage form at a rate that can replace the amount of drug that is metabolized and excreted from the body. Controlled release of an active ingredient can be stimulated by a variety of conditions including, but not limited to, pH, temperature, enzymes, water or other physiological conditions or compounds.

Parenteral dosage forms:

parenteral dosage forms can be administered to a patient by a variety of routes including, but not limited to, subcutaneous, intravenous (including single bolus intravenous injection), intramuscular, and intraarterial. Parenteral dosage forms are preferably sterile or capable of being sterilized prior to administration to a patient, since their administration will generally bypass the patient's natural defense system against contaminants. Examples of parenteral dosage forms include, but are not limited to, solutions ready for injection, dry products ready to be dissolved or suspended in a pharmaceutically acceptable injection carrier, suspensions and emulsions ready for injection.

Suitable carriers that can be used to provide the parenteral dosage forms of the invention are well known to those skilled in the art. Examples include, but are not limited to: water for injection USP; aqueous carriers such as, but not limited to, sodium chloride injection, ringer's injection, dextrose-sodium chloride injection, and lactated ringer's injection; water-miscible carriers such as, but not limited to, ethanol, polyethylene glycol, polypropylene glycol; and non-aqueous vehicles such as, but not limited to, corn oil, cottonseed oil, peanut oil, sesame oil, ethyl oleate, isopropyl myristate, and benzyl benzoate.

Compounds that increase the solubility of one or more of the active ingredients disclosed herein may also be included in the parenteral dosage forms of the invention.

Transdermal, topical and mucosal dosage forms:

transdermal, topical and mucosal dosage forms of the invention include, but are not limited to, ophthalmic solutions, sprays, aerosols, creams, lotions, ointments, gels, solutions, emulsions, suspensions or other forms known to those skilled in the art. See, e.g., Remington's Pharmaceutical Sciences (1980 & 1990) 16th and 18th eds., Mack Publishing, Easton, PA. and Introduction to Pharmaceutical document Forms (1985) 4th ed., Lea & Febiger, Philadelphia, Pa. Dosage forms suitable for use in treating mucosal tissue within the oral cavity may be formulated as mouthwashes or chewing gums (oral gels). In addition, transdermal dosage forms include "reservoir-type" patches or "matrix-type" patches that may be applied to the skin and held for a specified period of time to allow penetration of the desired amount of active ingredient.

Suitable excipients (e.g., carriers and diluents) and other materials that may be used to provide transdermal, topical and mucosal dosage forms encompassed by the present invention are well known to those skilled in the pharmaceutical arts and depend on the particular tissue to which a given pharmaceutical composition or dosage form is to be administered. Based on this fact, typical excipients include, but are not limited to, water, acetone, ethanol, ethylene glycol, propylene glycol, butane-1, 3-diol, isopropyl myristate, isopropyl palmitate, mineral oil, and mixtures thereof to form lotions, tinctures, creams, emulsions, gels or ointments, which are non-toxic and pharmaceutically acceptable. Humectants or wetting agents may also be added to the pharmaceutical compositions and dosage forms if desired. Examples of such additional ingredients are well known in the art. See, e.g., Remington's Pharmaceutical Sciences (1980 & 1990) 16th and 18th eds., Mack Publishing, E aston, Pa.

The pH of the pharmaceutical composition or dosage form or of the tissue to which the pharmaceutical composition or dosage form is administered may also be adjusted to improve delivery of one or more active ingredients. Similarly, the polarity of the solvent vehicle, its ionic strength, and tonicity can be adjusted to improve delivery. Compounds (e.g., stearates) can also be added to pharmaceutical compositions or dosage forms to advantageously alter the hydrophilicity or lipophilicity of one or more active ingredients to improve delivery. In this aspect, the stearate may serve as a lipid carrier for the formulation, as an emulsifier or surfactant, and as a delivery or permeation enhancer. Different salts, hydrates or solvates of the active ingredients may be used to further adjust the properties of the resulting composition.

Dose and frequency of administration:

can be used for treating and preventing NK₂The amount of a compound or composition of the invention that is a disorder or disease associated with receptor activity or one or more symptoms thereof will vary with the nature and severity of the disorder or disease and the route of administration of the active ingredient. The frequency and dosage will also vary depending on the particular factors for each patient, depending on the particular treatment (e.g., therapeutic or prophylactic) being administered; the severity of the disorder, disease or condition; the route of administration; and the age, weight, response, or past medical history of the patient. Effective doses can be extrapolated from dose response curves obtained from in vitro or animal model test systems. Suitable regimens may be selected by those skilled in the art by taking such factors into account and by following the dosages as reported in the literature and recommended in the physicians' Desk Reference (62 nd ed., 2008), for example.

Generally, the recommended daily dosage range for a compound of the invention for use in a disorder or disease described herein is in the range of about 0.01mg to about 2000mg per day, either as a single once-daily dose or as divided doses throughout the day. In one embodiment, the daily dose is administered twice daily in divided doses. Preferably, the daily dose ranges from about 5mg to about 1000mg per day, more particularly from about 10mg to about 500mg per day. In treating the patient, treatment should be initiated at lower doses, perhaps from about 1mg to about 25mg, and if necessary increased, up to about 200mg to about 1000mg daily, in single or divided doses, depending on the overall response of the patient. In some cases it may be desirable to use dosages of the active ingredient outside the ranges disclosed herein, as will be appreciated by those of ordinary skill in the art. Further, it should also be noted that the clinician or treating physician knows how and when to interrupt, adjust or terminate therapy in connection with individual patient responses.

Has been or is being used in therapy and NK₂Prophylactic or therapeutic agents other than the compounds of the present invention for disorders or diseases associated with receptor activity or one or more symptoms thereof may be used in the combination therapy of the present invention. For example, the compounds of the invention may be formulated with other antidepressants (e.g., antidepressants that inhibit the breakdown of 5-hydroxytryptamine, such as monoamine oxidase inhibitors). In one embodiment, the additional therapeutic agent is a therapeutic agent that binds to a glutamate receptor, such as an AMPA receptor, a kainate receptor, an agonist site for an NMDA receptor, or a glycine site for a strychnine insensitive NMDA receptor.

Examples of therapeutic agents that may be used to treat or prevent depression include, but are not limited to, tricyclic antidepressants, such as amitriptyline (amitriptyline), amoxapine (amoxapine), bupropion, clomipramine, desipramine (desipramine), doxepin (doxepin), imipramine, maprotiline (maprotiline), nefazadone (nefazadone), nortriptyline (nortriptyline), protriptyline (protriptyline), trazodone, trimipramine (trimipramine), and venlafaxine (venlafaxine); selective 5-hydroxytryptamine reuptake inhibitors such as fluoxetine (fluoxetine), fluvoxamine (fluvoxamine), paroxetine (parooxetine), and sertraline (sertraline); monoamine oxidase inhibitors such as isocarboxazid (isocarboxazid), eugenine (pargyline), phenelzine and tranylcypromine (tranylcypromine); and psychostimulants such as dextroamphetamine and methylpiperidine acetate.

Other examples of antidepressants that may be used include, but are not limited to, phenylindaparine (bindaline), acetominophenidone (caroxazone), citalopram (citalopram), diethyltheophylline (dimethazan), phendimine (fencamine), indaparine (indaparine), indolozoline hydrochloride (indoxacillin), nefopam (nefopam), phenformin (nomifensine), hydroxytryptamine (oxitriptolide), hydrastidine (oxyphylline), fenthiozolinone (thiazesim), benzazyl hydrazine (benmoxine), iprochlorohydrazide (iproclozide), iproniazin (ipronidazide), niazolidine (nialamidine), otaxine (octopamine), phenetyline, cotropinine (cotropinine), cyclidine (rolipramine), pyriproxerone (propanil), triamcinolone (benzathine), benzathine (azadirine), benzathinine (triamcinolone (benzathine), benzathine (benzathinine), benzathinine (triamcinolone (benzathine), benzathine (benzathine), benzathine (benzathine), benzathine, Benzothiepin (dothiepin), trifluoropropylamine (fluacizine), imipramine (N-oxide), aminocycloxindole (iprindole), lofepramine (lofepramine), tetrandrylamine (melitracen), metapramine (metapramine), oxitilin (norptin), etipramine (opipramol), pizotyline (pizotyline), propiconazole (propizepine), quinuclidine (quinuprazone), tianeptine (tiazetidine), aclonidine (adrafinil), benactyzine (benactyzine), butacetin (tabucin), diphenidine dioxane (dioxadolol), duloxetine (duloxetine), phentriadimerone (etolide), fenbamate (febuxate), mepiquin (oxyphenbutamine), oxyphenbutazine (oxyphenbutazine), oxyphenbutazine (oxyphenbutazone), oxyphenbutafenadine (oxyphenbutazone), oxyphenbutazone (oxyphenbutazone), fluphenazone (oxyphenbutazone (fenoxate), fluphenazone (oxyphenbutazone), fluphenazone (benoxazine), flubenoxazine (fluphenazone (fluzone), fluphenazone (flubenoxazine (fluphenazone (flubenoxazine), fluphenazone), fluben, Piribelamine (piperenaline), phenlopentane (prolinate), pirimidyl succinate (pyriruccinanol), ritanserin (ritanserin), rochellole (roxindole), rubidium chloride (rubidium chloride), sulpiride (sulpiride), tandospirone (tandospirone), fentezalone (thozalinone), benzethonium (tofenacin), toloxatone (toloxatone), L-tryptophan (L-trptophan), viloxazine (viloxazine), and phenylpyramine (zilidine).

Examples of therapeutic agents that may be used in the treatment or prevention of anxiety disorders include, but are not limited to, benzodiazepines (benzodiazepines), such as alprazolam (alprazolam), brotizololam (brotizololam), methotrexate (chlordiazepoxide), cloxapam (clobazam), clonazepam (clonazepam), chlordiazepam (clorazepam), desmethoxypam (demoxepam), diazepam (estazolam), flumazenil (flumazenil), flurazepam (flurazepam), halazepam (halazepam), lorazepam (lorazepam), midazolam (diazepam), nitrazepam (nitrazepam), nordiazepam (nozepam), oxazezezezepam (citazepam), pram (lorazepam), midazolam (diazepam), diazepam (fluazinazepam), terazepam (fluzeazepam), and diazepam (fluazinam (triazolam); non-benzodiazepines such as buspirone (buspirone), gepirone (gepirone), ixabepilone (ipsapirone), thiopyrrone (tiospirone), zopiclone (zolpirone), zolpidem (zolpidem), and zaleplon (zaleplon); neuroleptics such as barbiturates (barbiturates) such as amobarbital (amobarbital), aprebital (aprobarbital), butarbital (butarbital), isobarbital (butarbital), phenobarbital (mephobarbital), methohexital (methohexital), pentobarbital (pentobarbital), phenobarbital (phenobarbital), secobarbital (secobarbital), and thiopental (thiopental); and propylene glycol carbamates such as meprobamate (meprobamate) and tyloxaden (tybamate).

Examples of therapeutic agents that may be used to treat or prevent inflammatory bowel disease include, but are not limited to, anticholinergics, diphenoxylate (diphenoxylate), loperamide (loperamide), tincture of deodorized opium (tincture), codeine (codeine); broad spectrum antibiotics such as metronidazole, sulfasalazine, olsalazine, 5-aminosalicylic acid, prednisone, azathioprine, mercaptopurine, and methotrexate.

Examples of therapeutic agents useful for treating or preventing irritable bowel syndrome include, but are not limited to, propantheline (propantheline); muscarinic receptor antagonists such as pirenzapine (pirenzapine), methoctramine (methoctramine), ipratropium (ipratropium), tiotropium bromide (tiotropium), scopolamine (scopolamine), methscopolamine (methacocoamine), homatropine (homatropine), homatropine bromide (homatropine), and ethacrylene (methantheline); and antidiarrheals such as diphenoxylate (diphenoxylate) and loperamide (loperamide).

Examples of therapeutic agents that may be used to treat or prevent urinary incontinence include, but are not limited to, propofol (propantheline), imipramine, hyoscyamine, oxybutynin (oxybutynin), and dicyclomine.

Examples of therapeutic agents useful for treating or preventing inflammatory airway diseases include, but are not limited to, anti-inflammatory agents such as corticosteroids; a leukotriene modifier; mast cell stabilizers; and bronchodilators such as β -adrenergic agonists, drugs with anticholinergic effects and methylxanthines.

Preferably, the ratio is now used or is being used for treatment and NK₂Lower doses of a disorder or disease associated with receptor activity or one or more symptoms thereof may be used in the combination therapies of the invention. Are now used for the prevention, treatment, management or amelioration of NK₂The recommended dosage of an agent for a disorder or disease associated with receptor activity or one or more symptoms thereof may be obtained from any reference in the art, including, but not limited to, Hardman et al, eds.,1996, Goodman&Gilman's The Pharmacological Basis Of The therapeutics9.sup. th Ed, McGraw-Hill, New York, Physician's Desk Reference (PDR) 62nd Ed.,2008, Medical Economics Co., Inc., Montvale, NJ, which are incorporated herein by Reference in their entirety.

In some embodiments, when a compound of the invention is administered in combination with an additional therapeutic agent, the therapeutic agents (e.g., prophylactic or therapeutic agents) are administered simultaneously or separately, e.g., less than 30 minutes apart, 1 hour apart, 3 hours apart, 5 hours apart, 10 hours apart, 12 hours apart, 18 hours apart, 24 hours apart, 36 hours apart, 48 hours apart, or 72 hours apart.

Another aspect of the invention relates to a method for regulating NK₂A method of receptor activity comprising administering said NK₂The receptor is contacted with an effective amount of a compound of the invention.

NK₂The activity of the receptor can be increased or decreased (i.e., inhibited) by increasing or decreasing the NK₂The activity of the receptor is modulated. The NK₂The activity of the receptor can be reduced or inhibited by: an endogenous ligand that inhibits the receptor (e.g., the NK₂NKA of a receptor) or a commercially available exogenous ligand (e.g., saredutan) to the 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 NK₂The activity of the receptor may be reduced by 100% or less than 100% (e.g., 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, or 10%). For example, for NK₂Inhibition of receptor activity can occur by: formula (1) or formula(I) Binds to a binding site of an endogenous ligand, thereby reducing binding of the endogenous ligand. However, for NK₂Inhibition of receptor activity may also be achieved by binding a compound of formula (1) or formula (I) to NK₂The following sites on the receptor: this site is different from the endogenous ligand binding site, but is in contact with the NK₂Endogenous ligands of a receptor may still alter (e.g., decrease) their activity (e.g., allosteric modification of the receptor) upon interaction. The NK₂The activity of the receptor may be increased by 5% or more than 5% (e.g., 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80% or 90%, 100% or more than 100%). Regulation of NK₂The method of receptor activity can be performed in vitro (e.g., in a cell, a cell lysate, or a sample comprising a portion of a cell (e.g., only the relevant receptor)) or in vivo (e.g., in a human patient).

Other embodiments

The compounds of the invention can be used as research tools (e.g., to assess the mechanism of action of new agents, to isolate new drug discovery targets using affinity chromatography, as antigens in ELISA assays or ELISA-like assays, or as standards in vitro assays or in vivo assays). These and other uses and embodiments of the compounds and compositions of the present invention will be apparent to those of ordinary skill in the art.

The invention is further illustrated by reference to the following examples describing in detail the preparation of the compounds of the invention. It will be apparent to those skilled in the art that various modifications, both to materials and methods, may be practiced without departing from the purpose and meaning of the present invention. The following examples are given to aid in the understanding of the present invention and should not be construed as specifically limiting the invention described and claimed herein. Such variations of the invention, including all equivalents now known or later developed by those skilled in the art, as well as minor changes in formulation and experimental design, are intended to be within the scope of the invention.

Examples

Example 1

Preparation of fertilized egg isolate A

To produce fertilized egg isolate A, 8-9 day old whole fertilized chicken eggs 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 to or through a 1.0mm sterile screen. The egg shell and filtrate were discarded. The retentate, which contains the embryos, clear bursa and all or most of the egg white 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. Grinding the dried product with 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 deg.C (short term) or-20 deg.C (long term).

HPLC analysis

The final powder containing fertilized egg isolate a is analyzed by high performance (or High Pressure) Liquid Chromatography (HPLC). The results were quantified using a multi-wave absorption detector. The absorbance was read at 215 nm. A Pharmacia Superdex 20010/300GL molecular sieve column (10 mm 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. A representative chromatogram is shown in figure 1.

Verification of an analysis

The final powder containing isolate a was also subjected to standard analytical procedures to measure purity and levels of protein, fat, ash, moisture and various contaminants. The results for one representative sample are shown in figure 2.

Preparation A capsule

To prepare a capsule of formulation A, 4000.0g of the final powder containing isolate A, sodium benzoate (0.5% w/w) and potassium sorbate (0.2% w/w) were mixed with 40g of 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 2

Study of formulation A for treatment of Major Depressive Disorder (MDD) and its related disorders/symptoms

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 primary rating scale used in north america for assessing the degree of depression in patients. The overall score is explained 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 assesses 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.

Montgomery-Asneberg Depression rating Scale- "MADRS"

This is the primary rating scale used in north america for assessing the degree of depression in patients. 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 rating Scale, 'BDI'

This is a commonly used measure of depression, and is commonly used as a self-assessment tool. The total score is a simple sum of the scores of the 21 items. In general, a score of <9 indicates no or very mild depression, 10-18 indicates mild to moderate depression, 19-29 indicates moderate to severe depression, >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 exaggerated depression or a personality disorder of the performance 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 total score range available is 5-30, 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 assesses such problems as concentration ability, anxiety, poor confidence, poor self-value, unpleasantness, and depression. The scores were as follows:

the Likert Scale (Likert scale), from left to right, 0, 1,2, 3; 12 entries, each with a rating of 0-3, were evaluated.

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, indicating severe problems and psychological stress.

Diagnostic and Statistical Manual of Mental Disorders (Diagnostic and Statistical Manual of Mental Disorders) -IV th edition-revision text-DSM-IV TR "

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

The main effect measured was repeated analysis of variance (analysis of variance) with the score of HAM-D 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 label study was conducted at the west neilsberg Hospital (MSH) located in toronto, ontario, canada. Patients were recruited through media advertisements, MSH outpatient clinics, and other clinical center referrals.

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 such potential: significant improvement in MDD, above the level of known placebo effect well-demonstrated in other trials; and demonstrate that formulation a is an acceptable therapy in this patient population. The second objective of this pilot study was to evaluate the effect of formulation a on reducing anxiety symptoms and improving quality of life.

MDD was screened for each patient using DSM-IV TR criteria and HAM-D. Once they participated in the study, they were scheduled into an open label study of formulation a for 8 weeks. CGI strictness by a global measureThe patients were further evaluated on the Severe (GCI-S) and (CGI-I) improvement scales. The Udvalg for Kliniske is used for side effects(UKU) side effects rating Scale (Lingjaerde) was evaluated systematically. The second measure of depression symptoms is the montgomery-elsegberg depression rating scale (MADRS) and the becker depression rating scale (BDI) as a self-assessment tool. The quality of the life dimension is evaluated in a profile 36 (SF-36). Anxiety was assessed using 14 items of HAM-a.

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

Dosage of formulation A

The dose of formulation a was about 2000 mg/day (2 capsules of formulation a, each about 500mg, taken orally 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 for major depression (single episode or relapse) achieved DSM-IVTR criteria.

(ii) The 17 Hamilton depression rating Scale (HAM-D17) had a total score at baseline of 18 or more.

(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 based on the judgment of the clinician as being appropriate for the patient's standard of care. However, no enhancement strategy was allowed during the 8 week trial.

(iv) English reading and writing ability.

(v) A signed written informed consent was obtained,

(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 as having a significant risk of suicide (HAMD suicide score > 1), or a history showing a possible tendency to overt self-injury.

(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, breastfeeding, scheduled pregnancy in the next 12 months, or insufficiently protected women.

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

(vii) Course of electroconvulsive therapy (ECT) at the observation period.

(viii) Suffering from a severe 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 undergoing psychotherapy or starting psychotherapy in a trial.

(xii) Subjects with clinically significant abnormal laboratory results at blood screening and urinalysis.

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

Design of research

This is a single point and open label randomized study conducted in 23 patients (20 of them with analyzable results) designed to verify the efficacy and safety of formulation a monotherapy.

The trial consisted of an 8 week evaluation period, followed by a2 week antidepressant clearance period if necessary.

Screening

Once the physician and/or study coordinator adequately advises the subjects of the study, the nature of the treatment, and other choices they may make, and the subjects sign an informed consent document, the physician makes a clinical DSM IV TR diagnosis and administers HAM-D17. Qualified subjects were then reviewed for medical history, psychiatric history, and concurrent therapy, followed by physical examination. In addition, the study coordinator performed baseline laboratory tests including urine (Routine & Microscopic), CBC differential (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 and those with clinically significant abnormalities in laboratory tests were excluded.

Week 0

The patient was re-subjected to baseline examination (week 0) and prescribed a monotherapy with formulation a by the physician. The formulation a is administered to depressed and antidepressant patients who are taking antidepressants but have no effect.

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 determined at the physician clinician discretion. During this period, clearance of the patient was monitored by a psychiatric physician on a weekly check, and further monitored by the study coordinator at mid-weekly intervals via telephone. It is recognized that depression may worsen 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 major depressive exacerbation (decline) in this regimen to be substantially greater than with routine care, provided the subject is carefully monitored during this period 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 an excessive prolongation of depression time. 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 no different from standard of care with careful monitoring and administration of formulation a, a potentially effective agent. Moreover, as already discussed, standard care is only effective in about 60% of patients, thus often requiring the same possible re-assessment and drug adjustments.

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)

Montgomery-Asneberg Depression rating Scale (MADRS) (Montgomery)

-beck Depression rating Scale (BDI) (10)

Quality of life (SF-36) (Ware)

Hamilton anxiety rating Scale (HAMA) (Hamilton 1959)

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

Compliance with drugs (except V2)

Study examination estimated to take about 1 hour, except that the baseline 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 in the treatment of depression and the best clinical benefit of the patient.

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

Statistical method

The primary effect was tested using repeated 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 the following change in HAM-D17 with a standard deviation of 0.65 (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 detects an average change as small as 4.3 points on the 52 point scale with a weight (power) of 80%. Each participant had a HAM-D17 score greater than 17 according to the addition criteria. The Franck mitigation criterion is a HAM-D17 of 9 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 of responders and good responders (remitter) may be performed, if appropriate.

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 incomplete overall study in these 4 subjects included non-compliance with the dose and/or schedule, no patience with the results and subject's abroad.

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

Total score Table for 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 score table for 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

[0363] Total score 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

Total score Table for 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

Screening	Week 0	Week 2	Week 4	Examination of No. 7	Week 6	Week 8
							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

[0371] Total rating Scale 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. "responder" or "sustained responder" refers to 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 rate of decrease in HAM-D score was higher in the sustained responders who completed the 8 week study, 68.1%, a figure much higher than the minimum 50% decrease 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, because her score at this time is 12, which is only slightly worse than the 50% reduction compared to the score of 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 Extension Study (see example 3) where scores of 1, 11, 7 and 9 were recorded. Subject #106 faced considerable household confusion after the start of the extended study, 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 experienced improvement, and not everyone who experienced improvement maintained the improvement by 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 the treatment of major depressive disorder 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 3

The positive efficacy and safety results of the study described in example 2 required extensive studies. 10 subjects from the study described in example 2 participated in an extended study. The extended study was open only to those subjects in the study described in example 2 who were clinical responders at the end of the 8-week study. Formulation a was administered as described in example 1, and subjects of the extended study were analyzed monthly for 10 months. The following table shows the HAM-D scores for subjects in the extended study.

w/d = pull-out extension study

4 of the 10 subjects dropped out of the extended study due to the occurrence of exclusion criteria in the continued study. The results of this extended study indicated that all subjects in the study were responders to formulation a as defined. 6 out of 10 clinical responders (60%) were in remission at the start of the extended study. 8 of 10 subjects (80%) were in remission at the last day of evaluation. 2 of the subjects were clinical responders in the initial 8-week study, but did not improve until the end of the extended study. Only 1 subject (# 113) participating in the extended study as a clinical responder relapsed after participating in the extended study.

Example 4

The positive efficacy and safety results of the extended study described in example 3 required a second extended study. 4 subjects from the extended study described in example 3 participated in the second extended study. The second extended study was only open to those subjects from the extended study of example 3 who wished to continue taking formulation a. Formulation a was administered as described in examples 2 and 3. The second extended study plan lasted 12 months and 4 subjects entering the study now completed the study for 8 or 9 months. Each subject has been and will be analyzed every month. The following table shows the HAM-D scores of the subjects in the extended study.

Moon cake	Subject #102	Subject #106	Subject #110	Subject #123
					Examination No. 1	3	1	3	4
Examination No.2	0	3	0	2
					Examination No. 4	0	4	1	5
Examination No. 4	0	7	0
					Examination of 5 th	0	12	1
Examination of No.6	1	2	2
					Examination of No. 7	0	2	0
Examination of 8th	0		0
					Examination of 9 th	2
Examination of the 10 th time
					Examination of 11 th
Examination of 12 th

The results of this second extended study indicated that all subjects in this study (except subject #106 at the 5 th exam) remained in remission (i.e., had HAM-D scores of less than 8) for the entire period after they began entering the second extended study. All subjects were in remission at the last evaluation day.

Example 5

As described in the above examples, formulation a has been shown to have a therapeutic effect. Studies have been conducted to study the mechanism of action of formulation a, as described in PCT publication No. WO 2009/086634. In particular, studies were conducted to determine the inhibition of the binding interaction of the radioligand with its receptor, or the inhibition of 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) and the test was repeated. These concentrations of formulation a were prepared by: the capsule contents of formulation A were dissolved in dimethyl sulfoxide, and the solution was then 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 more than 60 different receptors and enzymes (as detailed in PCT publication No. WO 2009/086634). The mean percent inhibition of specific binding was then determined for each isolate a concentration.

Of the more than 60 receptors and enzymes tested, 5 receptors showed binding inhibitory activity. The results of this study indicate that neurokinin A interacts with human NK in the presence of isolate A (approximately 10. mu.g/mL)₂Receptor binding was inhibited by 32.15%. Separation constant (K)_d) Is 5x10^-10M, inhibition constant (K) of neurokinin A of reference compound_i) Is 2.53x 10^-10And M. In addition, the binding inhibition studies described above indicate that isolate a replaces 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 (approximately 10. mu.g/mL). Binding of radiolabeled kainate to the kainate receptor was inhibited by 22.38% in the presence of isolate A (approximately 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 (approximately 10. mu.g/mL). Radiolabelled MDL-105,519 was insensitive to strychnine in the NMDA receptor in the presence of isolate A (approximately 10. mu.g/mL)Binding of the sensitive glycine site was inhibited by 27.45%.

The NK is removed₂Receptors are used in additional receptor binding assays. A single concentration (one-concentration) controlled experiment was performed to assess the NK antagonism of various isolates of the capsule contents of formulation A₂The ability of the receptor to bind to the ligand. The contents of formulation a capsules were dissolved using a variety of solvents and extracted using 4 different methods, as detailed below. These extraction processes produce a variety of isolates. These isolates are referred to as: 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. To more easily track the binding activity in the assay, higher concentrations of the isolate (e.g., about 100. mu.g/ml) are used. The radioligand binding assay was performed based on the Burcher and Regoli method. Typically, recombinant human NK will be expressed₂A Chinese Hamster Ovary (CHO) cell of the receptor, in the presence of a control (neurokinin A) or each isolate¹²⁵I]Neurokinin A (final concentration 1.0 μm) was incubated. The reaction was carried out in the presence of 0.02% bovine serum albumin and 1mM MnCl₂At 25 ℃ for 4 hours in 20mM HEPES (pH 7.4). The reaction was then terminated by rapid vacuum filtration on a glass fiber filter. Measuring the radioactivity trapped on the filter and comparing to a control value to confirm that the isolate is in contact with the NK₂Any interaction of the neurokinin a binding site of the receptor (measured as a percentage of specific binding).

Sample #19 was prepared by weighing the contents of 103mg of formulation a capsules. Water (10.3 mL) was added and the solution was vortex mixed for 1 minute. 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 dried. The lower (aqueous) fraction was redissolved in 2.06mL of water. The sample was not clarified and centrifuged at 10,000rpm for 10 minutes in a microcentrifuge. The supernatant was removed and labeled as sample 085426-4 (sample #19 lower isolate) and used in receptor binding studies. The upper (organic) fraction was redissolved in 1.245mL of 20% aqueous acetonitrile. The sample was not clarified and centrifuged at 10,000rpm for 10 minutes in 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 capsule. 10mL of methanol dichloromethane (1: 1) was added and the solution was vortex mixed. 10mL of dichloromethane was then added to the solution, and the solution was again vortex mixed. 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 dried and redissolved in 2.49mL of 100% aqueous methanol. 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 was vortexed. The aqueous and organic fractions were 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 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 0.1 mg/mL), labeled as 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 receptor binding studies (note that this is the same control used for sample # 20).

Sample #2 was prepared as follows. One part (1.8 mg) of the contents of the formulation a capsule was weighed out. Then, 40% PEG in water (3.6 mL) with 0.25% Tween 80 (0.5 mg/mL) was added and the samples were 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 a receptor binding study.

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

Bold indicates over 50% inhibition at the tested concentration.

Fraction X isolate neurokinin A from its NK₂Binding of receptor inhibited 55%, sample #20 supernatant isolate neurokinin a with its NK₂Receptor binding was inhibited by 53%.

To further confirm NK₂Binding and activation of receptor sample #20 prepared as described aboveThe upper isolate was antagonistic and a dose response study was performed. Evaluation of NK in the presence of #20 supernatant (based on the amount of formulation A contained in sample #20 before extraction) at the following concentrations₂Inhibition of receptor binding: 0.1, 0.3, 1.0, 3.0, 10, 30, 100 and 300. mu.g/mL.

FIG.3 shows the expression of NK₂Results of assays performed by the receptor. Sample #20 supernatant inhibits neurokinin A binding to NK in a concentration-dependent manner₂Receptor capacity, the higher the concentration of the supernatant isolate of sample #20, the greater the inhibition of binding provided, and the lower the concentration, the less inhibition of binding provided. 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 #20₅₀Determined as 4.15x10²μg/mL，K_iDetermined as 3.49x10²M。

Example 6

And said NK₂The receptor interacting compounds are isolated as described below. A crude extract of formulation a (1.91 g, off-white amorphous) suspended in water (HPLC grade, j.t.baker) was loaded onto a WP C18 column (40 μm, j.t.baker). The packed column was eluted with water, 30% methanol, 85% methanol and 100% methanol (HPLC solvent, j.t. baker) injected in succession. Then, each aliquot of these methanol-containing fractions was applied to the human NK as described above₂Receptor radioligand binding assay. The results of this assay indicate that the maximum inhibition of binding occurs in the fractions eluting with 85% methanol and 100% methanol. When used at a concentration of 0.1mg/ml, both fractions exhibited an anti-NK of 98.2%₂Activity of (2). The methanol in the active fraction containing methanol (i.e. the fraction eluting with 85% methanol and 100% methanol) was removed under vacuum and the remaining water was removed by lyophilization. The dried fraction was stored at-20 ℃.

0.599g of active fraction (which is a combination of dried fractions eluted with 85% methanol and 100% methanol) which showed 98.2% of anti-human NK at 0.1mg/ml₂Receptor activity-loaded on a WP C18 column (Φ 2.1X 50cm,40 μm, J.T.Baker). By applying from 20%, 50% to 70% acetonitrile: (LC/MS grade, Fisher Scientific) multi-step gradient elution of the active fraction. The fraction of each aliquot (each fraction containing 10-15ml of eluate) in the human NK as described above₂And again in the receptor radioligand binding assay. Figure 4 shows the results for fractions 25, 51, 65, 115, 135, 155, 161, 171, 185, 191 and the control fraction, which contains only eluent (0.05% methanol). Fractions 171 and 185 showed 99.8% and 100.8% inhibition of binding of radiolabeled NKA, respectively. The solvent was removed and the dried sample fraction was stored at-20 ℃.

From 171 (99.8% anti-NK)₂) And 185 (100.8% anti NK)₂) The purity of most active fractions of (a) was characterized using HPLC-UV using standard conditions known to those skilled in the art. The results of these experiments are shown in figures 5-8. FIG.5 shows a chromatogram of fraction 171 and FIG.6 shows a chromatogram of fraction 185, both fractions being detected using light at 210 nm. FIG.7 shows a chromatogram of fraction 171 and FIG.8 shows a chromatogram of fraction 185, both fractions being detected using 190nm light.

600MHz¹H NMR (Bruker) was used to further evaluate fractions 171 and 185. These experiments show that both fractions are pure and both fractions contain the same compound. Fractions 170, 171, 172, 173 and 174 were combined into one sample using 600MHz¹H NMR HR mass spectrometry assay aimed at determining the compound (i.e., binding to NK) in this sample₂Compound of an acceptor).

One-dimensional NMR spectroscopic analysis shows that¹H-and¹³c resonance, which resolves a total of 14 protons (out of the final 36) and 18 carbons. NMR assignments for the protons are as follows

-4.23(1H, dd, 11.74, 4.70), 4.17(1H, dd, 11.74, 5.87), 3.95(1H, m), 3.72(1H, dd, 11.44, 4.11), 3.62(1H, dd, 11.44, 5.87), 2.37(2H, dd, 8.78, 7.62), 1.65(2H, m), 1.35-1.25 (overlap), 1.13(1H, m), 0.88(3H, overlap) and 0.86(3H, overlap); for¹³C-174.1(C＝O)，70.2(OCH)，65.1(OCH₂)，63.2(OCH₂)，36.5(CH₂)，34.3(CH)，34.0(CH₂)，29.9(CH₂)，29.5(CH₂)，29.5(CH₂)，29.4(CH₂)，29.3(CH2)，29.1(CH₂)，29.0(CH₂)，27.0(CH₂)，24.8(CH₂)，19.1(CH₃) And 11.3 (CH)₃)。

CH and CH₃The identification of (2) was confirmed by using the Distortionless Enhancement by Polarization Transfer (DEPT) NMR technique. DEPT-90 (middle) and-135 (upper) experiments also confirmed two CHs₃And two CH. Any positive peak in DEPT-135 that does not appear in DEPT-90 is CH₃。

Multidimensional NMR spectroscopy techniques are used to establish connectivity of the atomic and functional groups, which is used to elucidate the final two-dimensional structure.

Thus, these studies indicate that binding to the NK is possible₂The active compound of the receptor has the chemical structure depicted in formula (I), and the stereochemistry of the chiral center of the isolated compound is not determined.

First, heteronuclear single quantum correlation spectroscopy (HSQC, H-C) further confirmed CH, CH from one-dimensional experiments₂And CH₃And (4) identification of the unit.

The backbone connectivity of the glyceride units was determined by Double Quantum Filtering (DQFCOSY) and total correlation spectroscopy (TOCSY or homonuclear Hartmann Hahn spectroscopy, HOHAHA) experiments. DQFCOSY measures the connectivity of adjacent protons, while TOSCY can punch through several bonds.

Longer range linkages (other than glycerides) using heteronuclear phasesThe related art, namely HMBC (heteronuclear multiple bond correlation spectroscopy). This procedure allows glycerol (C1-H) to be linked to the carbonyl myristate (myristic carbonyl) group via a carboxyl (ester) linkage. This also provides the linkage between the C-alpha proton and the C-beta proton with the carbonyl group of the carboxyl function. The linkage of the C-alpha proton and the C-beta proton may also be extended to include some CH of myristic acid adjacent thereto₂A group.

The NMR analysis concluded that the NK was bound₂The compound of the receptor is identified as a compound of formula (I). The said assignment gives the molecular formula C₁₈H₃₆0₄The corresponding molecular weight is about 316, which is subsequently confirmed by AccuTOF experiments.

Example 7

2, 3-dihydroxypropyl 6-methyl-myristate was synthesized as described above and 2, 3-dihydroxypropyl myristate was obtained and dissolved in dimethyl sulfoxide (DMSO). Various concentrations of 2, 3-dihydroxypropyl 6-methyl-myristate and 2, 3-dihydroxypropyl myristate were tested against human NK described above in example 5₂Effect of receptor binding assay. FIG.9 shows the NK activity in the presence of 2, 3-dihydroxypropyl 6-methyl-myristate (labeled compound "107236-1" in FIG. 10)₂Results of assays performed on the receptor. In general, 6-methyl-2, 3-dihydroxypropyl myristate inhibits neurokinin A and the NK in a concentration-dependent manner₂The ability of the receptor to bind. IC of neurokinin A and 2, 3-dihydroxypropyl 6-methyl-myristate₅₀、K_iShown in fig. 9.

FIG.10 shows the NK effect in the presence of 2, 3-dihydroxypropyl myristate (labeled compound "107236-2" in FIG. 10)₂Results of radioligand binding assays performed at the receptor. The 2, 3-dihydroxypropyl myristate also inhibits neurokinin A and the NK₂The ability of the receptor to bind. IC of neurokinin A and 2, 3-dihydroxypropyl myristate₅₀、K_iShown in fig. 10.

Example 8

Synthesis of 2, 3-dihydroxypropyl 6-methyl-myristate was carried out as described above and gave 2, 3-dihydroxypropyl myristate. They were all prepared as solutions by adding dimethyl sulfoxide (DMSO) to a concentration of 10 mM. These compounds are then used in cellular/functional calcium flow agonist and antagonist assays to determine the compounds for human NK₂The effect of receptor activity, as measured by the change in the measured value of intracellular calcium. These assays were performed based on the method of Gerard et al.

Briefly, the NK₂Receptor agonist assays were performed as follows. Stably expressing recombinant human NK₂Recipient Chinese hamster ovary-K1 (CHO-K1) cells were placed on mixed extracellular matrix overnight in complete media. One hour prior to the assay, the medium was replaced with Hank's Buffered Salt Solution (HBSS) containing 0.1% bovine serum albumin. A dye for measuring intracellular calcium is then applied to these cells and a baseline measurement of intracellular calcium is taken. The control (concentration in the interval 1X 10)^-11M to 3x10^-7Agonists of M [ bAla⁸]-NKA (4-10)) or compounds (all at a concentration in the interval 3X10^-7M to 1x10^-4M2, 3-dihydroxypropyl myristate (compound 2) or 2, 3-dihydroxypropyl 6-methyl-myristate (compound 3)) was added to the appropriate cell wells. Fluorescence emission at 485nm excitation/515 nm was measured every two seconds for at least 2 minutes. The fluorescence peak height in each well receiving compound 2 or compound 3 was recorded and compared to the fluorescence peak height in wells receiving the control. The results of this assay are presented in FIG.11, which is [ bAla ]⁸]Curves of percentage maximum of NKA (4-10) control (% max response) versus compound concentration (log (compound) (M)) of control, 2, 3-dihydroxypropyl myristate (compound # 2) or 2, 3-dihydroxypropyl 6-methyl-myristate (compound # 3).

The NK₂Receptor antagonist assays were performed essentially as follows. Stably expressing recombinant human NK₂CHO-K1 cells of the recipient were placed on mixed extracellular matrix overnight in complete medium. One hour prior to the assay, the medium was replaced with hank's buffered saline (HBSS) containing 0.1% bovine serum albumin. A dye for measuring intracellular calcium is then applied to these cells and a baseline measurement of intracellular calcium is taken. The control (concentration in the interval 3X 10)^-8M to 1x10^-5Antagonist GR159897 or at a concentration of M in the interval 1x10^-11M to 3x10^-7[ bAla ] of M⁸]-NKA（4-10））（[bAla⁸]The calcium effect of NKA (4-10) disappears with time, thus, by blocking [ bAla ]⁸]Additional Effect of NKA (4-10) [ bAla⁸]NKAs (4-10) can act like antagonists in the antagonist assay) or samples (all at concentrations in the interval 3x10^-7M to 1x10^-4M2, 3-dihydroxypropyl myristate (compound 2) or 2, 3-dihydroxypropyl 6-methyl-myristate (compound 3)) was added to the appropriate cell wells. After 10 minutes, the agonist [ bAla ] was added⁸]NKA (4-10) (final concentration 0.3 nM). Fluorescence emission at 485nm excitation/515 nm was measured every two seconds for at least 2 minutes. The fluorescence peak height in each well receiving control, compound 2 or compound 3 was recorded and compared to the fluorescence peak height in wells receiving agonist only. The results of this experiment are presented in FIG.12, which is [ bAla ]⁸]Curves of percentage maximum of NKA (4-10) control (% max response) versus compound concentration (log (compound) (M)) of control, 2, 3-dihydroxypropyl myristate (compound # 2) or 2, 3-dihydroxypropyl 6-methyl-myristate (compound # 3). As shown in FIG.12, 2, 3-dihydroxypropyl myristate and 2, 3-dihydroxypropyl 6-methyl-myristate were responsible for this function NK₂Antagonistic activity was shown in both receptor antagonism assays.

It is believed that the preparation and use of the compounds of the present invention, including each of the compounds encompassed by formula 1, is clear from the foregoing description of exemplary embodiments and, thus, may be protected as such. It will be apparent to those skilled in the art that various changes and modifications can be made herein without departing from the spirit and scope of the invention.

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Claims (76)

1. A compound having the structure comprising a pharmaceutically acceptable salt of said compound:

wherein:

a and B are independently-OH or-SH,

v and W are independently oxygen or sulfur, and at least one of V and W is oxygen,

R₁is- (CH)₂)_pCH₃Or is-H, and

p is an integer of 0 to 3, and:

x is- (CH)₂)_m-，

Y is-H, and Y is-H,

z is- (CH)₂)_n-，

m and n are integers which are defined as,

m =1 to 5, and m is a linear chain,

n =4 to 14, and n is a linear variable,

for all m and n, 6. ltoreq. m + n. ltoreq.14,

wherein, optionally, there are up to two carbon-carbon double bonds, each double bond being formed between adjacent methylene groups of formula (1), wherein if there are two of said double bonds, each carbon atom thereof is bonded to at least one hydrogen;

or X is

Y is absent, C_AAnd C_BTogether form a double bond, and form a double bond,

z is- (CH)₂)_r-，

q and r are integers which are,

q =0 to 4, and q =0 to 4,

r =1 to 13, and r is a linear chain,

for all q and r, 5. ltoreq. q + r. ltoreq.13,

wherein, optionally, a second double bond is formed between adjacent methylene groups of formula (1) wherein each carbon atom thereof is bonded to at least one hydrogen;

or X is- (CH)₂)_t-，

Z is

Y is absent, C_AAnd C_cTogether form a double bond, and form a double bond,

R₁is- (CH)₂)_vCH₃Or is a compound of formula (I) or a compound of formula (II),

t and u are integers of which the number is,

t =1 to 5, and t is a linear variable,

u =0 to 12, and u =0,

for all t and u, 5. ltoreq. t + u. ltoreq.13,

wherein, optionally, a second double bond is formed between adjacent methylene groups of formula (1) wherein each carbon atom thereof is bonded to at least one hydrogen.

2. The compound of claim 1, wherein a and B are both-OH.

3. A compound according to claim 1 or 2 wherein V and W are both oxygen.

4. A compound according to any one of the preceding claims wherein R is₁Is- (CH)₂)_pCH₃。

5. A compound according to any one of the preceding claims wherein p is 0 to 2.

6. A compound according to any one of the preceding claims wherein p is 0 or 1.

7. A compound according to any one of the preceding claims wherein p is 0.

8. A compound according to any one of the preceding claims, wherein n =2 to 12 and 7 ≦ m + n ≦ 13.

9. A compound according to any one of the preceding claims, wherein n =3 to 11 and 8 ≦ m + n ≦ 12.

10. A compound according to any one of the preceding claims, wherein n =4 to 10 and 9 ≦ m + n ≦ 11.

11. The compound of any one of the preceding claims, wherein n =5 to 9 and m + n = 10.

12. The compound of any one of the preceding claims, wherein m =2 to 4.

13. The compound of any one of the preceding claims, wherein m = 3.

14. A compound according to any one of the preceding claims wherein r =2 to 12 and 6 ≦ q + r ≦ 12.

15. A compound according to any one of the preceding claims wherein r =3 to 11 and 7 ≦ q + r ≦ 11.

16. A compound according to any one of the preceding claims wherein r =4 to 10 and 8 ≦ q + r ≦ 10.

17. The compound of any one of the preceding claims, wherein r =5 to 9 and q + r = 9.

18. The compound of any one of the preceding claims, wherein q =1 to 3.

19. The compound of any one of the preceding claims, wherein q = 2.

20. A compound according to any one of the preceding claims, wherein u =1 to 11 and 6 ≦ t + u ≦ 12.

21. A compound according to any one of the preceding claims, wherein u =2 to 10 and 7 ≦ t + u ≦ 11.

22. A compound according to any one of the preceding claims, wherein u =3 to 9 and 8 ≦ t + u ≦ 10.

23. The compound of any one of the preceding claims, wherein u =4 to 8 and t + u = 9.

24. A compound according to any one of the preceding claims, wherein t =2 to 4.

25. The compound of any one of the preceding claims, wherein t = 3.

26. A compound according to any one of the preceding claims, wherein if said up to two carbon-carbon double bonds are present, each said bond is formed between methylene groups of Z.

27. The compound of claim 26, wherein said up to two carbon-carbon double bonds is one of said bonds.

28. A compound according to any one of the preceding claims, wherein if the second double bond is present, then the bond is formed between the methylene groups of Z.

29. A compound according to any one of the preceding claims, wherein no more than two carbon-carbon double bonds and no second double bond are present.

30. A compound of claim 1 having the formula:

31. a substantially stereochemically pure compound of claim 1 having the formula:

32. a substantially stereochemically pure compound of claim 1 having the formula:

33. a substantially stereochemically pure compound of claim 1 having the formula:

34. a substantially stereochemically pure compound of claim 1 having the formula:

35. a pharmaceutical composition comprising a compound according to any one of the preceding claims and a pharmaceutically acceptable carrier.

36. The pharmaceutical composition of claim 35, wherein the pharmaceutical composition is suitable for oral delivery, parenteral delivery, topical delivery, rectal delivery, vaginal delivery, administration by oral inhalation, or nasal delivery.

37. A dosage form comprising a compound of any one of claims 1-34.

38. The dosage form of claim 37, wherein the dosage form is a solution, suspension, syrup, tablet, capsule, microparticle, ointment, cream, or lozenge.

39. The dosage form of claim 38, wherein the dosage form is a capsule.

40. The dosage form of claim 39, wherein the dosage form is a tablet.

41. Therapeutic neurokinin 2 (NK)₂) A method of a disorder or disease associated with receptor activity, the method comprising the steps of: administering to a subject in need thereof a therapeutically effective amount of a compound of any one of claims 1-34, or a pharmaceutically acceptable salt thereof.

42. Therapeutic neurokinin 2 (NK)₂) A method of a disorder or disease associated with receptor activity, the method comprising the steps of: administering to a subject in need thereof a therapeutically effective amount of a compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein A and B are both-OH, V and W are oxygen, and R is₁Is H, X is- (CH)₂)_m-, Z is- (CH)₂)_n-, m is 3, n is 7, and the compound does not contain a C = C double bond.

43. The method of claim 41 or 42, wherein said and said NK₂The disorder or disease associated with receptor activity is a depressive mood disorder, an anxiety disorder, irritable bowel syndrome, inflammatory bowel disease, inflammatory airway disease or urinary incontinence.

44. The method of claim 43, wherein said and said NK₂The disorder or disease associated with receptor activity is a depressive mood disorder.

45. The method of claim 44, wherein said and said NK₂Correlation of receptor activityIs major depressive disorder.

46. The method of claim 45, wherein the subject is not undergoing psychotherapy treatment concurrently with the treatment.

47. The method of claim 45, wherein the subject is undergoing psychotherapy treatment concurrently with the treatment.

48. The method of any one of claims 41-47, wherein the compound is contained in a pharmaceutical formulation comprising a pharmaceutically acceptable carrier.

49. The method of any one of claims 41-48, wherein the step of administering a therapeutically effective amount of the compound further comprises administering another therapeutic agent.

50. The method of any one of claims 41-49, wherein the subject is a human.

51. A method for treating a disorder or syndrome associated with a depressive mood disorder, the method comprising the steps of: administering to a subject in need thereof a therapeutically effective amount of a compound of any one of claims 1-34, or a pharmaceutically acceptable salt thereof.

52. A method for treating a disorder or syndrome associated with a depressive mood disorder, the method comprising the steps of: administering to a subject in need thereof a therapeutically effective amount of a compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein A and B are both-OH, V and W are oxygen, and R is₁Is H, X is- (CH)₂)_m-, Z is- (CH)₂)_n-, m is 3, n is 7, and the compound does not contain a C = C double bond.

53. The method of claim 51 or 52, wherein the disorder or syndrome is a disorder of the brain or nervous system, an anxiety disorder, a sexual dysfunction, drug abuse, an eating disorder, or a hormonal disorder.

54. A method of treating a disorder or condition treatable by an antidepressant, the method comprising the steps of: administering to a subject in need thereof a therapeutically effective amount of a compound of any one of claims 1-34, or a pharmaceutically acceptable salt thereof.

55. A method of treating a disorder or condition treatable by an antidepressant, the method comprising the steps of: administering to a subject in need thereof a therapeutically effective amount of a compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein A and B are both-OH, V and W are oxygen, and R is₁Is H, X is- (CH)₂)_m-, Z is- (CH)₂)_n-, m is 3, n is 7, and the compound does not contain a C = C double bond.

56. The method of claim 54 or 55, wherein the disorder or condition treatable by an antidepressant is hot flashes associated with menopause, pain, or smoking cessation.

57. Used for adjusting NK₂A method of activity of a receptor comprising: bringing the NK₂Contacting the receptor with an effective amount of a compound according to any one of claims 1-34, or a pharmaceutically acceptable salt thereof.

58. Used for adjusting NK₂A method of activity of a receptor comprising: bringing the NK₂Contacting the receptor with an effective amount of a compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein A and B are both-OH, V and W are oxygen, and R is₁Is H, X is- (CH)₂)_m-, Z is- (CH)₂)_n-, m is 3, n is 7, and the compound does not contain a C = C double bond.

59. The method of claim 57 or 58, wherein said method is an in vivo method.

60. The method of claim 57 or 58, wherein said method is an in vitro method.

61. Use of a compound according to any one of claims 1 to 34, or a pharmaceutically acceptable salt thereof, for the treatment of a disorder or disease as defined in any one of claims 41 and 43 to 50.

62. Use of a compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein a and B are both-OH, V and W are oxygen, R is for the treatment of a disorder or disease as defined in any one of claims 41 and 43 to 50₁Is H, X is- (CH)₂)_m-, Z is- (CH)₂)_n-, m is 3, n is 7, and the compound does not contain a C = C double bond.

63. Use of a compound according to any one of claims 1 to 34, or a pharmaceutically acceptable salt thereof, for the treatment of a disorder or syndrome associated with a depressive mood disorder as defined in claims 51 or 53.

64. Use of a compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein a and B are both-OH, V and W are oxygen, R is-OH, for the treatment of a disorder or syndrome associated with depressive mood disorders as defined in claims 51 or 53₁Is H, X is- (CH)₂)_m-, Z is- (CH)₂)_n-, m is 3, n is 7, and the compound does not contain a C = C double bond.

65. Use of a compound according to any one of claims 1 to 34, or a pharmaceutically acceptable salt thereof, for the treatment of a disorder or condition treatable by an antidepressant, as defined in claim 54 or 56.

66. Use of a compound according to claim 1 or a pharmaceutically acceptable salt thereof for the treatment of a disease or disorder as defined in claim 54 or 56Use of a compound of formula (I) for the treatment of a disorder or condition treatable with a antidepressant, wherein A and B are both-OH, V and W are oxygen, and R is₁Is H, X is- (CH)₂)_m-, Z is- (CH)₂)_n-, m is 3, n is 7, and the compound does not contain a C = C double bond.

67. Use of a compound according to any one of claims 1 to 34, or a pharmaceutically acceptable salt thereof, for the modulation of NK as defined in any one of claims 57, 59 and 60₂Use of the activity of a receptor.

68. A compound according to claim 1 or a pharmaceutically acceptable salt thereof for use in the regulation of NK as defined in any one of claims 57, 59 and 60₂Use of the activity of an acceptor, wherein A and B are both-OH, V and W are oxygen, R₁Is H, X is- (CH)₂)_m-, Z is- (CH)₂)_n-, m is 3, n is 7, and the compound does not contain a C = C double bond.

69. Use of a compound according to any one of claims 1 to 34, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the treatment of a disorder or disease as defined in any one of claims 41 and 43 to 50.

70. Use of a compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein a and B are both-OH, V and W are oxygen, R is oxygen, for the manufacture of a medicament for the treatment of a disorder or disease as defined in any one of claims 41 and 43 to 50₁Is H, X is- (CH)₂)_m-, Z is- (CH)₂)_n-, m is 3, n is 7, and the compound does not contain a C = C double bond.

71. Use of a compound according to any one of claims 1 to 34, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the treatment of a disorder or syndrome associated with depressive mood disorders as defined in claims 51 or 53.

72. Use of a compound according to claim 1 or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the treatment of a disorder or syndrome associated with depressive mood disorders as defined in claim 51 or 53Wherein A and B are both-OH, V and W are oxygen, and R is₁Is H, X is- (CH)₂)_m-, Z is- (CH)₂)_n-, m is 3, n is 7, and the compound does not contain a C = C double bond.

73. Use of a compound according to any one of claims 1 to 34, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the treatment of a disorder or condition as defined in claim 54 or 56 which is treatable by an antidepressant.

74. Use of a compound according to claim 1, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the treatment of a disorder or condition treatable by an antidepressant as defined in claim 54 or 56 wherein A and B are both-OH, V and W are oxygen, and R is₁Is H, X is- (CH)₂)_m-, Z is- (CH)₂)_n-, m is 3, n is 7, and the compound does not contain a C = C double bond.

75. Use of a compound according to any one of claims 1 to 34, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for modulating NK as defined in claim 57 or 59₂Use of a receptor active agent.

76. Use of a compound according to claim 1 or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for modulating NK as defined in claim 57 or 59₂Use of a receptor active agent, wherein A and B are both-OH, V and W are oxygen, R₁Is H, X is- (CH)₂)_m-, Z is- (CH)₂)_n-, m is 3, n is 7, and the compound does not contain a C = C double bond.