Magnesium-containing oxytocin preparation and method of use

Abstract

To provide a magnesium-containing oxytocin preparation and a method of use. [Solution] Disclosed are methods and compositions for the treatment of autism spectrum disorder, related disorders, and symptoms of such disorders, comprising the simultaneous administration of oxytocin peptide and magnesium ions. The simultaneous administration of oxytocin peptide and magnesium ions results in a synergistic or enhanced effect on reducing social and communication deficits in patients with autism spectrum disorder. The present invention may address the need for oxytocin peptide formulations that can provide a more pronounced effect on the oxytocin treatment response in people with autism spectrum disorder and related disorders, which exhibit a wide range of variability.

Description

[Technical Field]

[0001] Cross-references to related applications This application claims priority under U.S. Provisional Patent Application No. 62 / 321,654, filed on April 12, 2016, the disclosure of which is incorporated herein by reference in its entirety.

[0002] Field of Invention The present invention relates to methods and compositions comprising oxytocin peptides and magnesium ions for the treatment of autism spectrum disorder, related disorders, and symptoms of such disorders. [Background technology]

[0003] (Background of the invention) Oxytocin is a naturally occurring 9-amino acid neuropeptide primarily produced in the paraventricular and supraoptic nuclei of the mammalian hypothalamus. Oxytocin is released into the central nervous system via dispersed neural pathways and into the peripheral circulation via the posterior pituitary gland. Intramuscular injection or intravenous infusion of synthetic oxytocin (Pitocin®) is currently approved in the United States to induce or improve uterine contractions to facilitate vaginal delivery and to manage postpartum bleeding. Intranasal oxytocin (Syntocinon®) was approved in the United States from 1960 to 1997 to stimulate milk ejection and promote breastfeeding. While the Syntocinon® nasal spray was withdrawn from the US market at the manufacturer's request, intranasal oxytocin is still sold in countries outside the US, such as Switzerland, Portugal, and Brazil. Recently, the use of oxytocin peptides in the treatment of autism spectrum disorder has been demonstrated. See WO2004 / 030524A2 and WO2008 / 042452A1 (these disclosures are incorporated herein by reference). Autism spectrum disorder is becoming increasingly prevalent in human populations and is typically recognized by certain behaviors and characteristics, such as impairments in communication skills and / or social interaction, lack of eye contact, and / or inability to form and / or maintain social relationships. Children and adults diagnosed with autism spectrum disorder may exhibit one or more of the aforementioned behaviors and characteristics to varying degrees. Common symptoms observed in individuals with autism spectrum disorder include persistent deficits in social communication and social interaction, social anxiety, and repetitive behaviors, restricted interests, and activities. Other behaviors and characteristics observed in individuals with autism spectrum disorder include avoidance of physical contact, generalized anxiety, monotonous or inability to regulate vocal volume, inability to develop peer relationships, lack of shared enjoyment and interests, and lack of social or emotional reciprocity. Other disorders that demonstrate deficits in social and communication include social anxiety disorder, obsessive-compulsive disorder, social (pragmatic) communication disorder, and neurodevelopmental disorders (including, but not limited to, attention deficit hyperactivity disorder, Prader-Willi syndrome, Timothy syndrome, fragile X syndrome, Rett syndrome, or Williams syndrome, which exhibit symptoms similar to those shown in autism spectrum disorder). Individuals with autism spectrum disorder tend to have communication deficits (e.g., responding inappropriately in conversation, misinterpreting nonverbal interactions, or having difficulty forming age-appropriate friendships). Furthermore, individuals with autism spectrum disorder may be overly reliant on established procedures, highly anxious and sensitive to changes in their environment, or intensely focused on inappropriate things (e.g., nanimate objects and / or narrow interests in specific topics). To reiterate, the symptoms of individuals with autism spectrum disorder are wide-ranging and continuous, with some individuals exhibiting mild symptoms and others severe symptoms. There are currently no available pharmacological treatments for core deficits in social communication and social interaction, or for restricted repetitive behaviors, interests, and activities in individuals with autism spectrum disorder and related disorders, and such treatments remain urgently needed. Oxytocin has been shown to improve core symptoms of autism, particularly social and communication deficits and associated anxiety symptoms. Clinical trials in humans have demonstrated the efficacy of intranasal oxytocin in treating autism spectrum disorder, related disorders, and symptoms of such disorders. See, for example, Yatawara et al., Mol. Psychiatry 2015, 1-9; Gorka et al., Neuropsychopharmacology 2015, 40(2):278-286; Anagnostou et al., Mol. Autism 2012, 3(1):16; Guastella et al., Psychoneuroendocrinology 2009, 34(6):917-923. However, these trials showed wide variability in the response of individuals with autism spectrum disorder and related disorders to needing oxytocin treatment. Therefore, there is a need for oxytocin peptide formulations that can provide more significant effects in the treatment of autism spectrum disorder and related disorders. [Prior art documents] [Patent Documents]

[0004] [Patent Document 1] International Publication No. 2004 / 030524 [Patent Document 2] International Publication No. 2008 / 042452 [Non-patent literature]

[0005] [Non-Patent Document 1] Yatawara et al., Mol. Psychiatry 2015, 1-9 [Non-Patent Document 2] Gorka et al., Neuropsychopharmacology 2015, 40(2):278-286 [Non-Patent Document 3] Anagnostou et al., Mol. Autism 2012, 3(1):16 [Non-Patent Document 4] Guastella et al., Psychoneuroendocrinology 2009, 34(6):917-923 [Overview of the project] [Means for solving the problem]

[0006] Brief summary of the invention Methods and compositions comprising oxytocin peptide and magnesium ions for the treatment of autism spectrum disorder, related disorders, and symptoms of such disorders are provided, the methods and compositions comprising the simultaneous administration of oxytocin peptide and magnesium ions via craniofacial mucosal administration (e.g., intranasal administration). The methods and magnesium-containing oxytocin peptide formulations described herein offer enhanced efficacy in treating autism spectrum disorder compared to oxytocin alone.

[0007] In one embodiment, the present invention provides a method for treating autism spectrum disorder, one or more symptomatic disorders associated with autism spectrum disorder, social and communication deficits, or anxiety, the method comprising the step of administering an effective dose of oxytocin peptide and magnesium ions to a subject in need, wherein the simultaneous administration of the oxytocin peptide and magnesium ions results in a synergistic or enhanced effect. The oxytocin peptide and magnesium ions may be administered together simultaneously or sequentially. In some embodiments, the oxytocin peptide is administered with the magnesium ions in the same unit dose or in separate unit doses or formulation units. In some embodiments, the oxytocin peptide and magnesium ions are administered sequentially. For example, the oxytocin peptide is administered a time after the administration of the magnesium ions. In some embodiments, the subject is human.

[0008] Oxytocin peptide and magnesium ions may be administered to subjects in need via the same route or via different routes. In some embodiments, oxytocin peptide is administered via craniofacial mucosal administration (e.g., intranasal, buccal, sublingual, or intraocular administration). In one embodiment, both oxytocin peptide and magnesium ions are administered intranasally as the same formulation.

[0009] In some embodiments, interleukin-6 (IL-6) is used as a biomarker of the potential effectiveness of oxytocin peptide administration in a subject for the treatment of autism spectrum disorder, a disorder presenting one or more symptoms associated with autism spectrum disorder, social and communication deficits, or anxiety; and for selecting a subject for application of the methods described herein. In some embodiments, the method includes the step of measuring the level of IL-6 in a subject, and the step of administering an effective dose of oxytocin peptide and magnesium ions to a subject having an elevated IL-6 level.

[0010] In one embodiment, a method for treating autism spectrum disorder, a disorder presenting one or more symptoms associated with autism spectrum disorder, social and communication deficits, or anxiety, the method comprising administering to a subject in need thereof an effective dose of oxytocin peptide and magnesium ions, wherein co-administration of the oxytocin peptide and the magnesium ions results in a synergistic or enhanced effect, further comprises administering to the subject an effective dose of interleukin-6 (IL-6), wherein administration of IL-6 further comprises inducing an increase in oxytocin receptor expression.

[0011] In some embodiments, the oxytocin peptide is human oxytocin consisting of Cys-Tyr-Ile-Gln-Asn-Cys-Pro-Leu-Gly (SEQ ID NO: 1). In some embodiments, the effective dosage of the oxytocin peptide is about 0.5 μg to about 2000 μg, preferably about 8 μg to about 1000 μg, more preferably about 15 μg to about 120 μg. In some embodiments, the magnesium ions of the above effective dosage to be administered are about 50 μg to about 68 mg, preferably about 50 μg to about 34 mg, more preferably about 1 mg to about 3 mg. In some embodiments, the above method includes a step of administering a magnesium salt (such as magnesium citrate and / or magnesium chloride) in an amount that provides about 50 μg to about 68 mg of magnesium, or about 50 μg to about 34 mg of magnesium, or about 1 mg to about 3 mg of magnesium. In some embodiments, the above method includes a step of administering magnesium citrate or magnesium chloride in an amount that provides about 50 μg to about 68 mg of magnesium, or about 50 μg to about 34 mg of magnesium, or about 1 mg to about 3 mg of magnesium. In some embodiments, the effective dosage of the oxytocin peptide and magnesium ions is about 0.5 μg to about 2000 μg or about 15 μg to about 120 μg (such as about 60 μg or about 66 μg) of the oxytocin peptide contained in an aqueous solution containing about 0.11% to about 2.8% (preferably about 1.1% to about 1.6%, for example, about 1.36%) (w / v) of magnesium, which is administered.

[0012] In some embodiments, the present invention provides a method for reducing one or more symptoms associated with autism spectrum disorder. Symptoms treatable by the above method include any social or communication defect treatable by an oxytocin peptide (such as a defect in eye contact, social anxiety, general anxiety, accuracy in determining social cues in a complex situation, empathy, and communication ability including expressive language function).

[0013] In some embodiments, the present invention provides methods for treating disorders exhibiting one or more symptoms associated with autism spectrum disorder. In some embodiments, the disorders include social anxiety disorder, obsessive-compulsive disorder, social (pragmatic) communication disorders, and neurodevelopmental disorders (including, but not limited to, attention deficit hyperactivity disorder, Prader-Willi syndrome, Timothy syndrome, fragile X syndrome, Rett syndrome, or Williams syndrome, which exhibit symptoms similar to those shown in autism spectrum disorder).

[0014] In one embodiment, the present invention provides a method for treating autism spectrum disorder, the method comprising the step of administering an effective dose of oxytocin peptide and magnesium ions (for example, intranasally) to a subject in need, wherein the simultaneous administration of the oxytocin peptide and magnesium ions results in a synergistic or enhanced effect. In one embodiment, the present invention provides a method for treating autism spectrum disorder, the method comprising the step of administering an effective dose of oxytocin peptide and magnesium ions to a subject in need, wherein the effective dose of the oxytocin peptide and magnesium ions is administered intranasally in a liquid formulation, and the volume of the liquid formulation administered is about 5 μL to about 1000 μL.

[0015] In one embodiment, the present invention provides a method for treating Prader-Willi syndrome, the method comprising the step of administering an effective dose of oxytocin peptide and magnesium ions (for example, intranasally) to a subject in need, wherein the simultaneous administration of the oxytocin peptide and magnesium ions results in a synergistic or enhanced effect. In one embodiment, the present invention provides a method for treating Prader-Willi syndrome, the method comprising the step of administering an effective dose of oxytocin peptide and magnesium ions to a subject in need, wherein the effective dose of oxytocin peptide and magnesium ions is administered intranasally in a liquid formulation, the volume of which is approximately 5 μL to approximately 1000 μL.

[0016] In one embodiment, a method is provided for treating deficiencies in sociality and communication, the method comprising the step of administering an effective dose of oxytocin peptide and magnesium ions (for example, intranasally) to a subject in need, wherein the simultaneous administration of the oxytocin peptide and magnesium ions results in a synergistic or enhanced effect. In one embodiment, the present invention provides a method for treating deficiencies in sociality and communication, the method comprising the step of administering an effective dose of oxytocin peptide and magnesium ions to a subject in need, wherein the effective dose of oxytocin peptide and magnesium ions is administered intranasally in a liquid formulation, the volume of which is approximately 5 μL to approximately 1000 μL.

[0017] In one embodiment, a method for treating anxiety is provided, comprising the step of administering an effective dose of oxytocin peptide and magnesium ions (for example, intranasally) to a subject in need, wherein the simultaneous administration of the oxytocin peptide and magnesium ions results in a synergistic or enhanced effect. In one embodiment, the present invention provides a method for treating anxiety, comprising the step of administering an effective dose of oxytocin peptide and magnesium ions to a subject in need, wherein the effective dose of oxytocin peptide and magnesium ions is administered intranasally in a liquid formulation, the volume of which is approximately 5 μL to approximately 1000 μL.

[0018] In some of these embodiments, the effective dose of the oxytocin peptide is about 0.5 μg to about 2000 μg. In some of these embodiments, the effective dose of the magnesium ion is about 50 μg to about 68 mg. In some of these embodiments, the effective dose of the oxytocin peptide and magnesium ion comprises about 15 μg to about 120 μg of oxytocin peptide administered in an aqueous solution containing about 1.1% to about 1.6% (w / v) magnesium. In some of these embodiments, the effective dose of the oxytocin peptide and magnesium ion comprises about 66 μg of oxytocin peptide administered in an aqueous solution containing about 1.36% magnesium. In some of these embodiments, the weight ratio of the administered oxytocin peptide to the administered magnesium ion is about 1:1 to about 1:1000. In some of these embodiments, the molar ratio of the administered oxytocin peptide to the administered magnesium ions is approximately 1:40 to approximately 1:40000. In some of these embodiments, the volume of the administered liquid formulation is approximately 50 μL to approximately 200 μL. In some of these embodiments, the liquid formulation is administered using a metered nasal spray (e.g., spray or puff) in 1 to 4 units of approximately 50 μL / unit. In some of these embodiments, the oxytocin peptide is human oxytocin (SEQ ID NO: 1).

[0019] In some of these embodiments, the liquid formulation is contained within a device for intranasal administration. In some of these embodiments, the intranasal administration device is a nasal pump device. In some of these embodiments, the nasal pump device comprises a storage bottle attached to a pump actuator. In some of these embodiments, the pump actuator measures to deliver a specific volume of approximately 50 μL. In some of these embodiments, the nasal pump device comprises a storage bottle attached to an aerozoizer. In some of these embodiments, the nasal pump device comprises one or more of the following: (i) a filter to prevent backflow, (ii) a metal-free channel, and (iii) a gamma-ray-stable plastic material.

[0020] Further provided are magnesium-containing oxytocin peptide formulations described herein for use in subjects in need of a method for treating autism spectrum disorder, one or more symptomatic disorders associated with autism spectrum disorder, social and communication deficits, or anxiety. Also provided are the use of magnesium-containing oxytocin peptide formulations described herein in the manufacture of pharmaceuticals for treating autism spectrum disorder, one or more symptomatic disorders associated with autism spectrum disorder, social and communication deficits, or anxiety.

[0021] Kits are also provided that include a magnesium-containing oxytocin peptide preparation as described herein, incorporated into an intranasal administration device such as a nasal pump, and suitable packaging materials. The kit may further include instructions for administering the magnesium-containing oxytocin peptide preparation to a subject in need for the treatment of autism spectrum disorder, one or more symptoms associated with autism spectrum disorder, social and communication deficits, or anxiety. In certain embodiments, for example, the following are provided: (Item 1) A method for treating autism spectrum disorder, one or more symptomatic disorders associated with autism spectrum disorder, social and communication deficits, or anxiety, the method comprising the step of administering an effective dose of oxytocin peptide and magnesium ions to a subject in need thereof, wherein the simultaneous administration of the oxytocin peptide and magnesium ions results in a synergistic or enhanced effect. (Item 2) The method according to item 1, wherein the oxytocin peptide is administered simultaneously with the magnesium ions. (Item 3) The method according to item 1, wherein the oxytocin peptide is administered before or after the administration of the magnesium ions. (Item 4) The oxytocin peptide is administered via craniofacial mucosal administration, according to any one of items 1 to 3. (Item 5) The method according to item 4, wherein the oxytocin peptide is administered via intranasal administration. (Item 6) The method according to item 5, wherein the oxytocin peptide and the magnesium ions are administered via intranasal administration. (Item 7) The method according to any one of items 1 to 6, wherein the effective dose of the oxytocin peptide is approximately 0.5 μg to approximately 2000 μg. (Item 8) The method according to any one of items 1 to 7, wherein the effective dose of the magnesium ions is approximately 50 μg to approximately 68 mg. (Item 9) The magnesium ion is provided using magnesium chloride and / or magnesium citrate, as described in any one of items 1 to 8. (Item 10) The method according to item 1, wherein the effective dose of the oxytocin peptide and the magnesium ions is approximately 15 μg to approximately 120 μg of the oxytocin peptide administered in an aqueous solution containing approximately 1.1% to approximately 1.6% (w / v) magnesium. (Item 11) The effective dose of the oxytocin peptide and the magnesium ions is the method according to item 1, having an oxytocin:magnesium molar ratio of about 1:40 to about 1:40000. (Item 12) The method described above is for treating autism spectrum disorder, and is the method described in any one of items 1 to 11. (Item 13) The method described above is for treating one or more disorders exhibiting symptoms associated with autism spectrum disorder, as described in any one of items 1 to 11. (Item 14) The aforementioned disorder is social anxiety disorder, obsessive-compulsive disorder, social (pragmatic) communication disorder, neurodevelopmental disorder, attention deficit hyperactivity disorder, Prader-Willi syndrome, Timothy syndrome, fragile X syndrome, Rett syndrome, or Williams syndrome, as described in item 13. (Item 15) The method described above is for addressing deficiencies in social skills and communication, and is the method described in any one of items 1 to 11. (Item 16) The method described above is for the purpose of treating anxiety, and is the method described in any one of items 1 to 11. (Item 17) The oxytocin peptide is human oxytocin (SEQ ID NO: 1), as described in any one of items 1 to 16. (Item 18) A method for treating autism spectrum disorder, one or more symptomatic disorders associated with autism spectrum disorder, social and communication deficits, or anxiety, the method comprising the step of administering an effective dose of oxytocin peptide and magnesium ions to a subject in need, wherein the effective dose of oxytocin peptide and magnesium ions is administered intranasally in a liquid formulation, the volume of the liquid formulation administered is approximately 5 μL to approximately 1000 μL. (Item 19) The effective dose of the oxytocin peptide is approximately 0.5 μg to approximately 2000 μg, as described in item 18. (Item 20) The effective dose of the magnesium ions is approximately 50 μg to approximately 68 mg, as described in item 18. (Item 21) The method according to item 18, wherein the effective dose of the oxytocin peptide and the magnesium ions comprises about 15 μg to about 120 μg of the oxytocin peptide administered in an aqueous solution containing about 1.1% to about 1.6% (w / v) magnesium. (Item 22) The effective dose of the oxytocin peptide and the magnesium ions is the method according to item 18, having an oxytocin:magnesium molar ratio of about 1:40 to about 1:40000. (Item 23) The method according to item 21 or 22, wherein the volume of the liquid formulation administered is approximately 50 μL to approximately 200 μL. (Item 24) The liquid formulation is administered using a metering nasal device in 1 to 4 units of approximately 50 μL / unit, according to the method described in item 23. (Item 25) The method described above is for treating autism spectrum disorder, as described in any one of items 18-24. (Item 26) The method described above is for treating one or more disorders exhibiting symptoms associated with autism spectrum disorder, as described in any one of items 18-24. (Item 27) The aforementioned disorder is social anxiety disorder, obsessive-compulsive disorder, social (pragmatic) communication disorder, neurodevelopmental disorder, attention deficit hyperactivity disorder, Prader-Willi syndrome, Timothy syndrome, fragile X syndrome, Rett syndrome, or Williams syndrome, as described in item 26. (Item 28) The method described above is for addressing deficiencies in social skills and communication, as described in any one of items 18-24. (Item 29) The method described above is for the purpose of treating anxiety, as described in any one of items 18-24. . (Item 30) The oxytocin peptide is human oxytocin (SEQ ID NO: 1), as described in any one of items 18 to 29. (Item 31) The liquid formulation is contained within a device for intranasal administration, as described in item 18. (Item 32) The method according to item 31, wherein the device for intranasal administration is a nasal pump device. (Item 33) The nasal pump device according to item 32, wherein the nasal pump device comprises a storage bottle attached to a pump actuator. (Item 34) The pump actuator meters to deliver a specific volume of approximately 50 μL, as described in item 33. (Item 35) The nasal pump device according to item 32, wherein the nasal pump device comprises a storage bottle attached to an aerozoizer. (Item 36) The nasal pump device described above is as follows: (i) Filters to prevent backflow, (ii) Metal-free channels, and (iii) Plastic materials that are stable against gamma rays The method described in any one of items 32 to 35, comprising one or more of the following: (Item 37) A composition comprising an oxytocin peptide and magnesium ions, wherein the oxytocin peptide and magnesium ions are present in amounts that produce a synergistic or enhanced effect when used to treat anxiety. (Item 38) The composition according to item 37, wherein the oxytocin peptide is human oxytocin (SEQ ID NO: 1). (Item 39) The composition according to item 37, wherein the composition is a liquid formulation containing about 0.01 mg / mL to about 16 mg / mL of the oxytocin peptide. (Item 40) The composition according to item 37, wherein the composition is a liquid formulation containing magnesium in an amount that provides approximately 3 mg / mL to approximately 30 mg / mL of magnesium. (Item 41) The composition according to item 37, wherein the oxytocin peptide and the magnesium ions have a molar ratio of about 1:40 to about 1:40000. (Item 42) The composition according to item 41, wherein the molar ratio is approximately 1:40 to approximately 1:800. (Item 43) The composition according to item 41, wherein the molar ratio is approximately 1:800 to approximately 1:40000. (Item 44) A composition according to any one of items 37 to 43, further comprising a device for craniofacial mucosal administration. (Item 45) The composition according to item 44, wherein the oxytocin peptide and the magnesium ions are contained within the device for craniofacial mucosal administration. (Item 46) The device is for intranasal administration, and is the composition described in item 45. [Brief explanation of the drawing]

[0022] [Figure 1]Figure 1 shows the effects of saline, oxytocin, magnesium citrate, and oxytocin combinations, as well as magnesium citrate, in a rat model of social behavior.

[0023] [Figure 2] Figure 2 shows the effects of saline, oxytocin, magnesium citrate, and oxytocin combinations, as well as magnesium citrate, in a rat model of anxiety.

[0024] [Figure 3] Figures 3A and 3B show the effects of magnesium citrate, oxytocin, and combinations of magnesium citrate and oxytocin in an elevated plus-maze rat model of anxiety. [Modes for carrying out the invention]

[0025] Detailed description of the invention The present invention provides, in particular, a method for treating autism spectrum disorder, one or more symptomatic disorders associated with autism spectrum disorder, social and communication deficits, or anxiety in subjects in need by craniofacial administration (e.g., intranasal administration) of oxytocin peptide and magnesium ions, or magnesium-containing oxytocin peptide formulations described herein. The oxytocin peptide and magnesium ions are administered in an effective dose that produces a synergistic or enhanced effect compared to the administration of oxytocin peptide alone.

[0026] (definition) As used herein, “oxytocin peptide” refers to a substance having biological activity related to natural oxytocin. Oxytocin peptides may be naturally occurring endogenous peptides, their fragments, analogs, or derivatives. Oxytocin peptides may also be non-endogenous peptides, their fragments, analogs, or derivatives. In one aspect, oxytocin peptide is human oxytocin. In other aspects, oxytocin peptide may be an analog or derivative of human oxytocin.

[0027] As used herein, “analog” or “derivative” means any peptide that is similar to naturally occurring oxytocin in which one or more amino acids are substituted, deleted, or inserted. The term also means any peptide in which one or more amino acids (e.g., one, two, or three amino acids) are modified, for example, by chemical modification. In general, the term encompasses all peptides that exhibit oxytocin activity but may have different potency or pharmacological profiles, if desired.

[0028] As used herein, unless otherwise specified, the terms “treatment” or “to treat” mean an approach to obtain a beneficial or desired outcome (e.g., a clinical outcome). With respect to autism spectrum disorder and related disorders, beneficial or desired outcomes include, but are not limited to, symptom reduction and / or reduction in the severity of symptoms, e.g., social and / or communication deficits and / or reduction of repetitive behaviors and / or anxiety. Social and communication deficits may include, but are not limited to, impairments in communication skills and / or social interaction, lack of eye contact and / or inability to form and / or maintain social relationships.

[0029] "Cooperative action," "synergistic action," or "synergistic effect" refers to the combined action of two or more compounds in a manner in which one compound complements or enhances the action of another, resulting in a greater effect than could be predicted or anticipated by summing the effects of two or more compounds administered individually in given doses. A "synergistic effect" is considered achieved when the combined use of two or more drugs produces a greater overall effect (e.g., improvement in social and communication deficiencies and / or reduction of anxiety) than could be predicted or anticipated by summing the effects of any one of the individual drugs at equal doses. Similarly, a "synergistic effect" is considered achieved when the combined use of two or more drugs results in an earlier onset and / or longer-lasting effect than would occur after administration of the individual drugs used alone at equal doses.

[0030] "Craniofacial mucosal administration" refers to delivery to the mucosal surface of the nose, nasal tract, and nasal cavity; the mucosal surface of the oral cavity including the gingiva (gums), floor of the mouth, lips, tongue, sublingual oral surface (including the lingual frenulum and floor of the mouth); the mucosal surface of the eye or the mucosal surface around the eye (including the conjunctiva, lacrimal gland, and nasolacrimal duct); and the upper or lower eyelid and mucous membrane of the eye.

[0031] "Intranasal administration" or "administered into the nasal cavity" means delivery to the nose, nasal tract, or nasal cavity by means of spray, droplets, powder, gel, film, inhalant, or other means.

[0032] The "lower region of the nasal cavity" generally refers to the part of the nasal cavity where the middle and inferior turbinates protrude and which is significantly innervated by the trigeminal nerve. The "upper region of the nasal cavity" is defined by the upper third and the cribriform region, where the olfactory innervation is located.

[0033] When used herein, "subject" or "patient" means a mammal, including but not limited to humans. Mammals include, but are not limited to, livestock (e.g., cattle), sports animals, pets (e.g., guinea pigs, cats, dogs, rabbits, and horses), primates, mice, and rats. In one embodiment, the subject is a human.

[0034] It should be noted that, as used herein, the singular forms "a," "an," and "the" refer to multiple objects unless otherwise indicated. Furthermore, as used herein, the terms "comprising" and their cognates are used in their comprehensive sense; that is, they are equivalent to the terms "including" and their corresponding cognates.

[0035] Where a range of values ​​is provided, each value that lies between the upper and lower limits of that range, and any other specified or intermediate values ​​within that range, are intended to be included within the scope of this disclosure. For example, if the range is specified as 1 μg to 8 μg, then 2 μg, 3 μg, 4 μg, 5 μg, 6 μg, and 7 μg, as well as the range of values ​​greater than 1 μg and less than 8 μg, are also intended to be expressly disclosed. If the range is specified as 10 to 14%, then 10%, 11%, 12%, 13%, and 14% are also intended to be expressly disclosed. Furthermore, each narrower range within a specified range, between any specified or intermediate value and any other specified or intermediate value, is included within the scope of this disclosure. These narrower upper and lower limits may be independently included in or excluded from the scope, and each scope in which one or both limits are included in the narrower scope, or each scope in which neither limit is included in the narrower scope, is included in the disclosure, constrained by any limit explicitly excluded from that defined scope. If the defined scope includes one or both limits, the scope excluding either or both of those included limits is also included in the disclosure.

[0036] (Oxytocin peptide) Oxytocin was one of the first peptide hormones to be isolated and sequenced. Natural oxytocin is a nine-amino acid cyclic peptide hormone with two cysteine ​​residues forming a disulfide crosslink between positions 1 and 6. The amino acid sequence of human oxytocin is Cys-Tyr-Ile-Gln-Asn-Cys-Pro-Leu-Gly (SEQ ID NO: 1).

[0037] Processes for producing oxytocin have been reported. See, for example, U.S. Patent Nos. 2,938,891 and 3,076,797. Furthermore, oxytocin is commercially available. Various peptide analogs and peptide derivatives are available, and others may be intended for use in the present invention, produced according to known methods, and tested for biological activity. Oxytocin analogs may include, but are not limited to, compounds in the related series containing ortho-trigluoro-ethoxyphenylacetyl cores, such as 4-threonine-1-hydroxy-deaminooxytocin, 4-serine-8-isoleucine-oxytocin, 9-deaminooxytocin, 7-D-proline-oxytocin and its deamino analogs, (2,4-diisoleucine)-oxytocin, deaminooxytocin analogs, 1-deamino-1-monocarba-E12-Tyr(OMe)-OT(dCOMOT), 4-threonine-7-glycine-oxytocin (TG-OT), oxypressin, deamino-6-carba-oxytoxin (dC60), L-371,257, and L-374,943.Other exemplary oxytocin analogs include 4-threonine-1-hydroxy-deaminooxytocin, 9-deaminooxytocin, an oxytocin analog containing a glycine residue instead of a glycinamide residue, (2,4-diisoleucine)-oxytocin, an oxytocin analog with natriuretic and diuretic activity, deaminooxytocin analog; long-acting oxytocin analog, 1-deamino-1-monocarba-E12-[Tyr(OMe)]-OT(dCOMO T), carbetocin, (1-butanoic acid-2-(O-methyl-L-tyrosine)-1-carbaoxytocin, deamino-1-monocarba-(2-O-methyltyrosine)-oxytocin [d(COMOT)]), [Thr4-Gly7]-oxytocin (TG-OT), oxypressin, Ile-conopressin, deamino-6-carbaoxytoxin (dC60), d[Lys(8)(5 / 6C-fluorescein)]VT, d[Thr(4),Lys(8)(5 / 6C-fluorescein)]V T, [HO(1)][Lys(8)(5 / 6C-fluorescein)]VT, [HO(1)][Thr(4),Lys(8)(5 / 6C-fluorescein)]VT, d[Om(8)(5 / 6C-fluorescein)]VT, d[Thr(4),Om(8)(5 / 6C-fluorescein)]VT, [HO(1)][Om(8)(5 / 6C-fluorescein)]VT, [HO(1)][Thr(4),Om(8)(5 / 6C-fluorescein)]VT and 1-deaminooxytocin (residues 1 and 6 Examples include desamino-oxytocin analogs (where the disulfide bridge between them is replaced with a thioether) and desamino-oxytocin analogs (where the disulfide bond is replaced with a diselenide bond, diterlide bond, telluloseleno bond, tellulosulfide bond, or selenosulfide bond) (for example, the peptide analog of oxytocin described in PCT patent application WO2011 / 120,071, which is incorporated herein by reference). The peptides for use in the present invention may be peptides obtained in naturally occurring peptide sequences or by partial substitution, addition, or deletion of amino acids in natural peptide sequences.The peptides can be chemically modified, for example, by amidation of the carboxyl terminus (-NH2), the use of D amino acids in the peptide, the incorporation of small non-peptidyl moieties, and modification of the amino acids themselves (e.g., alkylation or esterification of the side-chain R group). Such analogs, derivatives, and fragments should substantially retain the desired biological activity of the natural oxytocin peptide. In some embodiments, the oxytocin analog is 4-serine-8-isoleucine-oxytocin or 9-deaminooxytocin. In some embodiments, the oxytocin analog is carbetocin. This disclosure also includes other known oxytocin analogs, e.g., peptidogenic oxytocin receptor agonists described in PCT patent application WO2012 / 042371 and Wisniewski et al., J Med Chem. 2014, 57:5306-5317 (the entire contents of these are incorporated herein by reference). In some embodiments, the oxytocin analog is Wisniewski et al., J. The compounds are selected from compound numbers 1 to 65 listed in Tables 1 to 3 in Med Chem. 2014, 57: 5306-5317. In some embodiments, the oxytocin analog is selected from the group consisting of compound number 31 ([2-ThiMeGly7]dOT), compound number 47 (carba-6-[Phe2,BuGly7]dOT), compound number 55 (carba-6-[3-MeBzlGly7]dOT), and compound number 57 (carba-1-[4-FBzlGly7]dOT, also known as melotocin).

[0038] In some embodiments, oxytocin or oxytocin analogs are isotope-labeled by having one or more atoms replaced by isotopes having different atomic masses. Examples of isotopes that may be incorporated into the disclosed compounds include isotopes of hydrogen (e.g., 2 H and 3 H), carbon isotopes (e.g., 13 C and 14 C), nitrogen isotopes (for example, 15N), isotopes of oxygen (e.g., 18 O and 17 O), isotopes of phosphorus (e.g., 31 P and 32 P), isotopes of fluorine (e.g., 18 F), isotopes of chlorine (e.g., 36 Cl) and isotopes of sulfur (e.g., 35 S). Isotopically labeled compounds can be administered to a subject or other subject according to conventional techniques and then detected, providing useful diagnostic and / or therapeutic management data. Further, isotopically labeled compounds can be administered to a subject or other subject as needed, providing therapeutically beneficial absorption, distribution, metabolism and / or excretion profiles. All isotopic variations of oxytocin peptides, such as human oxytocin or its analogs or derivatives, are contemplated, whether radioactive or not.

[0039] In some embodiments, the oxytocin peptide is human oxytocin consisting of Cys-Tyr-Ile-Gln-Asn-Cys-Pro-Leu-Gly (SEQ ID NO: 1).

[0040] The International Unit (IU, UI, or IE) is an internationally accepted unit of activity used to quantify vitamins, hormones, and vaccines. The IU defines the amount of a substance that gives a unit of activity measured using a prescribed biological assay to standardize preparations derived from multiple raw materials. Similarly, the USP unit is a prescribed dosage unit established by the United States Pharmacopeia in cooperation with the U.S. Food and Drug Administration to ensure the identity, potency, quality, purity, and consistency of drug products. Generally, the USP unit is equivalent to the IU due to an attempt at coordination. By convention, in the case of oxytocin, one unit of activity is usually defined as being equivalent to approximately 2 micrograms of synthetic oxytocin peptide; or 1 mg is equivalent to 500 units (Stedman's Medical Dictionary). Thus, as used herein, 1 "IU" or 1 "International Unit" of oxytocin peptide is the amount of oxytocin peptide that has the same biological activity as approximately 2 micrograms of synthetic peptide or produces the same level of biological action (e.g., a contraction response in rat uterine tissue). If the analog is less active, more of the substance will be required to achieve the same level of biological effect. Measurement of drug potency is well known to those skilled in the art and may include either an in vitro or in vivo assay using synthetic oxytocin as a reference. Atke and Vilhardt Acta Endocrinol 1987:115(1):155-60; Engstrom et al., Eur J Pharmacol 1998:355(2-3):203-10.

[0041] (Magnesium-containing oxytocin peptide preparation) In a method of the present invention for treating autism spectrum disorder, one or more symptomatic disorders associated with autism spectrum disorder, social and communication deficits, or anxiety, comprising the step of administering an effective dose of oxytocin peptide and magnesium ions to a subject in need thereof, the oxytocin peptide and magnesium ions may be administered in a magnesium-containing oxytocin peptide formulation or composition. In one embodiment, the magnesium-containing oxytocin peptide formulation or composition contains oxytocin peptide and magnesium ions in an amount that produces a synergistic or enhanced effect when used in the treatment of autism spectrum disorder, one or more symptomatic disorders associated with autism spectrum disorder, social and communication deficits, or anxiety.

[0042] The relative ratio of oxytocin peptide to magnesium ions in magnesium-containing oxytocin peptide preparations is crucial in achieving optimal synergistic or enhanced effects. The optimal amounts of oxytocin peptide and magnesium ions may depend on other factors such as the specific disorder or symptom, the desired type of synergistic or enhanced effect, and the route of administration. For example, the amount of magnesium may be important for a faster onset of effect; the amount of oxytocin may be important for a longer-lasting effect; and the relative ratio of oxytocin to magnesium may be important for achieving maximum improvement in social functioning, reduction of social and communication deficits, and / or reduction of anxiety.

[0043] In some embodiments, the magnesium-containing oxytocin peptide preparation or composition is a liquid preparation containing about 0.01 mg / mL to about 16 mg / mL of oxytocin peptide. In some embodiments, the amount of oxytocin peptide in the liquid preparation is greater than about (lower limit) 0.01 mg / mL, 0.05 mg / mL, 0.1 mg / mL, 0.2 mg / mL, 0.3 mg / mL, 0.4 mg / mL, 0.5 mg / mL, 1 mg / mL, or 2 mg / mL. In some embodiments, the amount of oxytocin peptide in the liquid preparation is less than about (upper limit) 16 mg / mL, 12 mg / mL, 10 mg / mL, 8 mg / mL, 6 mg / mL, 4 mg / mL, 2 mg / mL, 1.6 mg / mL, 1.2 mg / mL, 1 mg / mL, 0.8 mg / mL, 0.6 mg / mL, 0.4 mg / mL, 0.3 mg / mL, 0.2 mg / mL, or 0.1 mg / mL. In other words, the amount of oxytocin peptide in the liquid formulation is within a range of approximately 0.01 to 16 mg / mL, with the lower limit being less than the upper limit.In some embodiments, magnesium-containing oxytocin peptide preparations or compositions are available in concentrations of approximately 0.01 mg / mL to approximately 12 mg / mL, approximately 0.05 mg / mL to approximately 16 mg / mL, approximately 0.1 mg / mL to approximately 12 mg / mL, approximately 0.1 mg / mL to approximately 8 mg / mL, approximately 0.1 mg / mL to approximately 4 mg / mL, approximately 0.1 mg / mL to approximately 2 mg / mL, approximately 0.1 mg / mL to approximately 1.6 mg / mL, approximately 0.1 mg / mL to approximately 1.2 mg / mL, and approximately 0.1 mg / mL to approximately 1 mg / mL, about 0.1 mg / mL to about 0.8 mg / mL, about 0.1 mg / mL to about 0.4 mg / mL, about 0.1 mg / mL to about 0.3 mg / mL, about 0.2 mg / mL to about 16 mg / mL, about 0.2 mg / mL to about 12 mg / mL, Approximately 0.2 mg / mL to approximately 10 mg / mL, approximately 0.2 mg / mL to approximately 8 mg / mL, approximately 0.2 mg / mL to approximately 6 mg / mL, approximately 0.2 mg / mL to approximately 4 mg / mL, approximately 0.2 mg / mL to approximately 2 mg / mL, approximately 0.2 mg / mL to approximately 1. 6mg / mL, about 0.2mg / mL to about 1.2mg / mL, about 0.2mg / mL to about 1mg / mL, about 0.2mg / mL to about 0.8mg / mL, about 0.2mg / mL to about 0.6mg / mL, about 0.2mg / mL to about 0.4mg / m L, about 0.2 mg / mL to about 0.3 mg / mL, about 0.3 mg / mL to about 16 mg / mL, about 0.3 mg / mL to about 12 mg / mL, about 0.3 mg / mL to about 10 mg / mL, about 0.3 mg / mL to about 8 mg / mL, about 0.3 mg / mL The oxytocin peptide contains oxytocin peptides in concentrations of approximately 4 mg / mL to 0.3 mg / mL to 0.3 mg / mL, 0.3 mg / mL to 0.1 mg / mL, 0.3 mg / mL to 0.5 mg / mL, 0.5 mg / mL to 0.16 mg / mL, 0.5 mg / mL to 0.10 mg / mL, 0.5 mg / mL to 0.5 mg / mL to 0.5 mg / mL, 0.5 mg / mL to 0.1 mg / mL, 1 mg / mL to 0.16 mg / mL, 1 mg / mL to 0.10 mg / mL, or 1 mg / mL to 0.5 mg / mL. In preferred embodiments, the magnesium-containing oxytocin peptide preparation or composition contains oxytocin peptides in concentrations of approximately 0.1 mg / mL to 0.2 mg / mL, 0.15 mg / mL to 0.15 mg / mL, or 0.2 mg / mL to 0.2 mg / mL.In one embodiment, the oxytocin peptide is human oxytocin consisting of Cys-Tyr-Ile-Gln-Asn-Cys-Pro-Leu-Gly (SEQ ID NO: 1).

[0044] In some embodiments, the magnesium-containing oxytocin peptide formulation or composition is a liquid formulation containing oxytocin peptide in an amount of about 5 IU / mL to about 8000 IU / mL. In some embodiments, the amount of oxytocin peptide in the liquid formulation is greater than about (lower limit) 5 IU / mL, 25 IU / mL, 50 IU / mL, 75 IU / mL, 100 IU / mL, 150 IU / mL, 200 IU / mL, 250 IU / mL, 500 IU / mL, 750 IU / mL, or 1000 IU / mL. In some embodiments, the amount of oxytocin peptide in the liquid formulation is less than approximately (upper limit) 8000 IU / mL, 6000 IU / mL, 5000 IU / mL, 4000 IU / mL, 3000 IU / mL, 2000 IU / mL, 1000 IU / mL, 800 IU / mL, 600 IU / mL, 500 IU / mL, 400 IU / mL, 300 IU / mL, 200 IU / mL, 150 IU / mL, 100 IU / mL, or 50 IU / mL. That is, the amount of oxytocin peptide in the liquid formulation is in any range of approximately 5 to 8000 IU / mL, with the lower limit being less than the upper limit.In some embodiments, magnesium-containing oxytocin peptide formulations or compositions are available in concentrations of approximately 500 IU / mL to approximately 6000 IU / mL, approximately 25 IU / mL to approximately 8000 IU / mL, approximately 50 IU / mL to approximately 6000 IU / mL, approximately 50 IU / mL to approximately 4000 IU / mL, approximately 50 IU / mL to approximately 2000 IU / mL, approximately 50 IU / mL to approximately 1000 IU / mL, approximately 50 IU / mL to approximately 800 IU / mL, approximately 50 IU / mL to approximately 600 IU / mL, approximately 50 IU / mL to approximately 500 IU / mL, and approximately 50 IU / mL~about 400IU / mL, about 50IU / mL~about 200IU / mL, about 50IU / mL~about 150IU / mL, about 100IU / mL~about 8000IU / mL, about 100IU / mL~about 6000IU / mL, about 100IU / mL~about 5000 IU / mL, about 100IU / mL to about 4000IU / mL, about 100IU / mL to about 3000IU / mL, about 100IU / mL to about 2000IU / mL, about 100IU / mL to about 1000IU / mL, about 100IU / mL to about 800IU / mL, about 1 00IU / mL~about 600IU / mL, about 100IU / mL~about 500IU / mL, about 100IU / mL~about 400IU / mL, about 100IU / mL~about 300IU / mL, about 100IU / mL~about 200IU / mL, about 100IU / mL~about 15 0IU / mL, about 150IU / mL to about 8000IU / mL, about 150IU / mL to about 6000IU / mL, about 150IU / mL to about 5000IU / mL, about 150IU / mL to about 4000IU / mL, about 150IU / mL to about 2000IU / mL, Contains oxytocin peptides in concentrations of approximately 150 IU / mL to 1500 IU / mL, 150 IU / mL to 500 IU / mL, 150 IU / mL to 250 IU / mL, 250 IU / mL to 8000 IU / mL, 250 IU / mL to 5000 IU / mL, 250 IU / mL to 2500 IU / mL, 250 IU / mL to 500 IU / mL, 500 IU / mL to 8000 IU / mL, 500 IU / mL to 5000 IU / mL, or 500 IU / mL to 2500 IU / mL.In preferred embodiments, the magnesium-containing oxytocin peptide preparation or composition contains an oxytocin peptide in a concentration of approximately 50 IU / mL to approximately 1000 IU / mL, approximately 75 IU / mL to approximately 750 IU / mL, or approximately 100 IU / mL to approximately 600 IU / mL. In one embodiment, the oxytocin peptide is human oxytocin consisting of Cys-Tyr-Ile-Gln-Asn-Cys-Pro-Leu-Gly (SEQ ID NO: 1).

[0045] The amount of magnesium present in the above preparation is expressed as weight percentage (w / v) (magnesium or Mg per 100 mL of solution). 2+ (grams), mg / mL (magnesium or Mg per milliliter of solution) 2+ (milligrams) or molar concentration (magnesium or Mg per liter of solution) 2+ "M" is defined as the number of moles; or magnesium or Mg per liter of solution. 2+ It can also be expressed as "mM" (defined as millimoles).

[0046] In some embodiments, the magnesium-containing oxytocin peptide preparation or composition contains approximately 1 mg / mL to approximately 30 mg / mL of magnesium or magnesium ions (Mg 2+The liquid formulation contains ) ). In some embodiments, the composition contains about 11 mg / mL to about 15 mg / mL of magnesium or magnesium ions. In some embodiments, the amount of magnesium or magnesium ions in the liquid formulation is greater than about (lower limit) 1 mg / mL, 2 mg / mL, 3 mg / mL, 4 mg / mL, 5 mg / mL, 6 mg / mL, 7 mg / mL, 8 mg / mL, 9 mg / mL, 10 mg / mL, 11 mg / mL, or 12 mg / mL. In some embodiments, the amount of magnesium or magnesium ions in the liquid formulation is less than about (upper limit) 30 mg / mL, 25 mg / mL, 20 mg / mL, 15 mg / mL, 14 mg / mL, 13 mg / mL, 12 mg / mL, 11 mg / mL, 10 mg / mL, 9 mg / mL, 8 mg / mL, 7 mg / mL, 6 mg / mL, or 5 mg / mL. That is, the amount of magnesium or magnesium ions in the liquid formulation is any of the ranges from about 1 to 30 mg / mL, with the lower limit being less than the upper limit. In some embodiments, the magnesium-containing oxytocin peptide preparation or composition comprises about 0.01 mg / mL to about 16 mg / mL (preferably about 0.1 mg / mL to about 2 mg / mL, more preferably about 0.15 mg / mL to about 1.5 mg / mL, or about 0.33 mg / mL) of oxytocin peptide and about 1 mg / mL to about 30 mg / mL (or about 3 mg / mL to about 30 mg / mL, about 4 mg / mL to about 30 mg / mL, about 5 mg / mL to about 30 mg / mL, about 8 mg / mL to about 30 mg / mL, about 10 mg / mL to about 30 mg / mL, preferably about 11 mg / mL to about 15 mg / mL, or about 13 mg / mL, or about 12 mg / mL) of magnesium or Mg 2+ The liquid formulation contains [a certain substance]. In some embodiments, the above magnesium-containing oxytocin peptide formulation or composition contains about 50 mM to about 1500 mM magnesium or magnesium ions (Mg 2+The liquid formulation contains magnesium or magnesium ions. In some embodiments, the amount of magnesium or magnesium ions in the liquid formulation is greater than approximately (lower limit) 50 mM, 100 mM, 150 mM, 200 mM, 250 mM, 300 mM, 350 mM, 400 mM, 450 mM, 500 mM, 550 mM, or 600 mM. In some embodiments, the amount of magnesium or magnesium ions in the liquid formulation is less than approximately (upper limit) 1500 mM, 1200 mM, 1000 mM, 750 mM, 700 mM, 650 mM, 600 mM, 550 mM, 500 mM, 450 mM, 400 mM, 350 mM, 300 mM, or 250 mM. That is, the amount of magnesium or magnesium ions in the liquid formulation is any of the ranges from approximately 50 to 1500 mM, with the lower limit being less than the upper limit. In some embodiments, the magnesium-containing oxytocin peptide preparation or composition comprises about 5 IU / mL to about 8000 IU / mL (preferably about 50 IU / mL to about 1000 IU / mL, more preferably about 75 IU / mL to about 750 IU / mL, or about 150 IU / mL) of oxytocin peptide and about 1 mg / mL to about 30 mg / mL (preferably about 11 mg / mL to about 15 mg / mL, or about 13 mg / mL, or about 12 mg / mL) of magnesium or Mg 2+ The liquid formulation contains the following. In some embodiments, the magnesium-containing oxytocin peptide formulation or composition contains about 5 IU / mL to about 8000 IU / mL (preferably about 50 IU / mL to about 1000 IU / mL, more preferably about 75 IU / mL to about 750 IU / mL, or about 150 IU / mL) of oxytocin peptide and about 50 mM to about 1200 mM (or about 100 mM to about 1200 mM, about 150 mM to about 1200 mM, about 200 mM to about 1200 mM, about 300 mM to about 1200 mM, about 400 mM to about 1200 mM, preferably about 400 mM to about 600 mM, or about 500 mM) of magnesium or Mg 2+ It is a liquid formulation containing [ingredient].

[0047] Any magnesium salt (e.g., a water-soluble magnesium salt) may be used to provide magnesium ions in a magnesium-containing oxytocin peptide formulation. The magnesium salt used in the above magnesium-containing oxytocin peptide formulation may be selected based on several factors (e.g., the amount of free magnesium ions that can be delivered when the formulation is administered, the solubility of the magnesium salt in the medium for the liquid formulation, the acidity / basicity of the counterion, and / or the dissociation constant of the salt). For example, in a liquid formulation, the magnesium salt needs to be sufficiently soluble in the liquid medium to deliver magnesium ions at the concentration required to produce a synergistic or enhanced effect with the oxytocin peptide. Other factors (e.g., compatibility with other substances in the formulation, and the ability of the counterion to perform other functions in the formulation) may also be considered when selecting a magnesium salt. For example, magnesium citrate is sufficiently soluble in aqueous solution to provide a desired amount of magnesium or a desired magnesium ion concentration; the citrate may be pharmaceutically acceptable; the citrate may be part of a buffer; and magnesium citrate may add a desirable flavor to the formulation. Magnesium ions in magnesium-containing oxytocin peptide formulations may be provided by using one or more magnesium salts. The magnesium salt in a magnesium-containing oxytocin peptide formulation may be the first magnesium salt used in the preparation of the formulation, or it may be formed in situ during the preparation of the formulation. For example, magnesium chloride may be the first magnesium salt used in the preparation of the formulation; magnesium citrate may be formed in situ when citric acid is added to the formulation. In such cases, magnesium ions in a magnesium-containing oxytocin peptide formulation are provided by both magnesium chloride and magnesium citrate.

[0048] In some embodiments, the magnesium-containing oxytocin peptide preparation or composition comprises one or more magnesium salts selected from the group consisting of magnesium citrate, magnesium chloride, magnesium sulfate, magnesium acetate, magnesium lactate, magnesium stearate, magnesium oxide, magnesium carbonate, magnesium glycinate, magnesium malate, magnesium taurate, magnesium gluconate, magnesium succinate, and magnesium pyrophosphate. In some embodiments, the magnesium-containing oxytocin peptide preparation or composition is a liquid preparation containing a magnesium salt (e.g., magnesium citrate or magnesium chloride) in an amount that provides about 1 mg / mL to about 30 mg / mL of magnesium. In some embodiments, the composition contains about 1 mg / mL to about 30 mg / mL of magnesium ions (Mg 2+ The magnesium salt is contained in an amount that provides ) the magnesium-containing oxytocin peptide preparation or composition, which contains about 1 mg / mL to about 30 mg / mL of magnesium or magnesium ions (Mg 2+ The liquid formulation contains one or more magnesium salts (e.g., magnesium citrate and / or magnesium chloride) in an amount that provides about 11 mg / mL to about 15 mg / mL of magnesium or magnesium ions. In some embodiments, the composition contains one or more magnesium salts in an amount that provides about 11 mg / mL to about 15 mg / mL of magnesium or magnesium ions. In some embodiments, the magnesium-containing oxytocin peptide formulation or composition contains about 1 mg / mL to about 30 mg / mL of magnesium or Mg 2+The liquid formulation contains an oxytocin peptide and a magnesium salt (e.g., magnesium citrate or magnesium chloride) in an amount of about 0.01 mg / mL to about 16 mg / mL (preferably about 0.1 mg / mL to about 2 mg / mL, more preferably about 0.15 mg / mL to about 1.5 mg / mL or about 0.33 mg / mL) in an amount that provides the desired result. In some embodiments, the magnesium-containing oxytocin peptide formulation or composition contains about 1 mg / mL to about 30 mg / mL (preferably about 11 mg / mL to about 15 mg / mL or about 13 mg / mL or about 12 mg / mL) of magnesium or Mg 2+ The liquid formulation contains an oxytocin peptide in an amount of about 5 IU / mL to about 8000 IU / mL (preferably about 50 IU / mL to about 1000 IU / mL, more preferably about 75 IU / mL to about 750 IU / mL or about 150 IU / mL) and one or more magnesium salts (e.g., magnesium citrate and / or magnesium chloride). In some embodiments, the magnesium-containing oxytocin peptide formulation or composition contains about 50 mM to about 1200 mM (or about 100 mM to about 1200 mM, about 150 mM to about 1200 mM, about 200 mM to about 1200 mM, about 300 mM to about 1200 mM, about 400 mM to about 1200 mM, preferably about 400 mM to about 600 mM or about 500 mM) of magnesium or Mg 2+ The liquid formulation contains an oxytocin peptide in an amount of about 5 IU / mL to about 8000 IU / mL (preferably about 50 IU / mL to about 1000 IU / mL, more preferably about 75 IU / mL to about 750 IU / mL or about 150 IU / mL) and one or more magnesium salts (e.g., magnesium citrate and / or magnesium chloride).

[0049] The relative amounts of oxytocin peptides and magnesium ions in magnesium-containing oxytocin peptide formulations or compositions detailed herein may be defined by weight ratio or by molar ratio. The weight ratio of the amount of oxytocin peptide to the amount of magnesium or magnesium ions in a formulation or composition is referred to as the "OT / Mg(w) ratio". For example, in a magnesium-containing oxytocin peptide formulation or composition having an OT / Mg(w) ratio of about 1:40, for every 1 mg of oxytocin peptide present in the formulation or composition, there is about 40 mg of magnesium or magnesium ions present in the formulation or composition. The molar ratio of the amount of oxytocin peptide to the amount of magnesium or magnesium ions in a formulation or composition is referred to as the "OT / Mg(m) ratio". For example, in a magnesium-containing oxytocin peptide formulation or composition having an OT / Mg(m) ratio of about 1:1600, for every 1 μmol of oxytocin peptide present in the formulation or composition, there is about 1600 μmol of magnesium or magnesium ions present in the formulation or composition.

[0050] In some embodiments, the magnesium-containing oxytocin peptide formulation or composition has an OT / Mg(w) ratio of about 1:1 to about 1:1000. In some embodiments, the OT / Mg(w) ratio in the formulation or composition is less than about (upper limit) 1:1, 1:2, 1:5, 1:10, 1:20, 1:30, 1:40, 1:45, 1:50, 1:60, 1:80, 1:100, or 1:200. In some embodiments, the OT / Mg(w) ratio in the formulation or composition is greater than about (lower limit) 1:1000, 1:800, 1:500, 1:250, 1:200, 1:150, 1:100, 1:80, 1:60, 1:50, 1:40, 1:30, 1:20, 1:10, or 1:5. That is, the OT / Mg(w) ratio in the above formulation or composition is any of the ranges from about 1:1 to 1:1000, with the upper limit being greater than the lower limit. In some embodiments, the formulation or composition has an OT / Mg(w) ratio of about 1:2 to about 1:200. In some preferred embodiments, the formulation or composition has an OT / Mg(w) ratio of about 1:30, about 1:35, about 1:40, about 1:45, or about 1:50. In some embodiments, the formulation or composition is approximately 1:2 to 1:1000, approximately 1:2 to 1:800, approximately 1:2 to 1:500, approximately 1:2 to 1:250, approximately 1:2 to 1:150, approximately 1:2 to 1:100, approximately 1:2 to 1:80, approximately 1:2 to 1:60, approximately 1:2 to 1:50, approximately 1:2 to 1:40, approximately 1:2 to 1:30, approximately 1:2 to 1:20, approximately 1:2 to 1:10, approximately 1:2 to 1:5, approximately 1:5 to 1:1000, approximately 1:5 to 1:800, approximately 1:5 to 1:500, approximately 1:5 to 1:200, approximately 1: 5~approx. 1:100, approx. 1:5~approx. 1:80, approx. 1:5~approx. 1:60, approx. 1:5~approx. 1:50, approx. 1:5~approx. 1:40, approx. 1:5~approx. 1:30, approx. 1:5~approx. 1:20, approx. 1:5~approx. 1:10, approx. 1:10~approx. 1:1000, approx. 1:10~approx. 1:800, approx. 1:10~approx. 1:500, approx. 1:10~approx. 1:200, approx. 1:10~approx. 1:10, approx. 1:10~approx. 1:80, approx. 1:10~approx. 1:60, approx. 1:10~approx. 1:50, approx. 1:10~approx. 1:40, approx. 1:10~approx. 1:30, approx. 1:10~approx. 1:20, approx. 1:20~approx. 1:1000,Approximately 1:20~1:800, Approximately 1:20~1:500, Approximately 1:20~1:200, Approximately 1:20~1:100, Approximately 1:20~1:80, Approximately 1:20~1:70, Approximately 1:20~1:60, Approximately 1:20~1:50, Approximately 1:20~1:40, Approximately 1:20~1:30, Approximately 1:30~1:1000, Approximately 1:30~1:800, Approximately 1:30~1:500, Approximately 1:30~1:200, Approximately 1:30~1:100, Approximately 1:30~ 1:80, approximately 1:30-1:70, approximately 1:30-1:60, approximately 1:30-1:50, approximately 1:30-1:40, approximately 1:35-1:45, approximately 1:40-1:1000, approximately 1:40-1:800, approximately 1:40-1:500, approximately 1:40-1:200, approximately 1:40-1:100, approximately 1:40-1:80, approximately 1:40-1:70, approximately 1:40-1:60, approximately 1:40-1:50, approximately 1:50-1:1000, approximately 1:50 ~approximately 1:800, approximately 1:50~approximately 1:500, approximately 1:50~approximately 1:200, approximately 1:50~approximately 1:100, approximately 1:50~approximately 1:90, approximately 1:50~approximately 1:80, approximately 1:50~approximately 1:70, approximately 1:50~approximately 1:60, approximately 1:60~approximately 1:100, approximately 1:60~approximately 1:80, approximately 1:60~approximately 1:80, approximately 1:60~approximately 1:50, approximately 1:60~approximately 1:200, approximately 1:60~approximately 1:100, approximately 1:60~approximately 1:90, approximately 1:60~approximately 1:80, approximately 1:60~approximately 1:70, It has an OT / Mg(w) ratio of approximately 1:80 to 1:1000, approximately 1:80 to 1:800, approximately 1:80 to 1:500, approximately 1:80 to 1:200, approximately 1:80 to 1:100, approximately 1:100 to 1:1000, approximately 1:100 to 1:800, approximately 1:100 to 1:500, approximately 1:100 to 1:200, approximately 1:200 to 1:1000, approximately 1:200 to 1:800, approximately 1:200 to 1:500, or approximately 1:500 to 1:1000. In one embodiment, the oxytocin peptide is human oxytocin consisting of Cys-Tyr-Ile-Gln-Asn-Cys-Pro-Leu-Gly (SEQ ID NO: 1), and / or its magnesium salt is magnesium citrate.

[0051] In some embodiments, the magnesium-containing oxytocin peptide formulation or composition has an OT / Mg(m) ratio of about 1:40 to about 1:40,000. In some embodiments, the OT / Mg molar ratio in the formulation or composition is less than about (upper limit) 1:40, 1:80, 1:100, 1:150, 1:175, 1:200, 1:250, 1:280, 1:300, 1:400, 1:500, 1:560, 1:800, 1:1000, 1:1100, 1:1200, 1:1600, 1:1700, 1:1800, 1:2000, 1:2400, 1:3200, 1:4000, or 1:8000. In some embodiments, the OT / Mg molar ratio in the above formulation or composition is greater than approximately (lower limit) 1:40000, 1:30000, 1:20000, 1:10000, 1:7500, 1:5000, 1:4000, 1:3000, 1:2500, 1:2000, 1:1600, 1:1200, 1:1100, 1:1000, 1:800, 1:600, 1:400, or 1:200. That is, the OT / Mg(w) ratio in the above formulation or composition is any of the ranges from approximately 1:40 to 1:40000, with the upper limit being greater than the lower limit. In some embodiments, the above formulation or composition has an OT / Mg(m) ratio of approximately 1:80 to approximately 1:8000. In some preferred embodiments, the above formulations or compositions have an OT / Mg(m) ratio of about 1:175, about 1:280, about 1:560, about 1:1100, about 1:1200, about 1:1400, about 1:1600, about 1:1700, about 1:1800, or about 1:2000. In some embodiments, the above formulations or compositions have an OT / Mg(m) ratio of about 1:80 to about 1:40000, about 1:80 to about 1:30000, about 1:80 to about 1:20000, about 1:80 to about 1:10000, about 1:80 to about 1:7500, about 1:80 to about 1:5000, about 1:80 to about 1:3000, about 1:80 to about 1:2000, or about 1 :80~approx. 1:1600, approx. 1:80~approx. 1:1200, approx. 1:80~approx. 1:800, approx. 1:80~approx. 1:400, approx. 1:80~approx. 1:200, approx. 1:175~approx. 1:40000, approx. 1:175~approx. 1:30000, approx. 1:175~approx. 1:20000, approx. 1:175~approx. 1:10000, approx. 1:175~approx. 1:5000,Approximately 1:175~1:3000, Approximately 1:175~1:2400, Approximately 1:175~1:2000, Approximately 1:175~1:1700, Approximately 1:175~1:1600, Approximately 1:175~1:1200, Approximately 1:175~1:1100, Approximately 1:175~1:800, Approximately 1:17 5~approx. 1:560, approx. 1:175~approx. 1:400, approx. 1:175~approx. 1:280, approx. 1:200~approx. 1:40000, approx. 1:200~approx. 1:30000, approx. 1:200~approx. 1:20000, approx. 1:200~approx. 1:10000, approx. 1:200~approx. 1:5000, approx. 1:200~approx. 1 :3000, approx. 1:200~approx. 1:2400, approx. 1:200~approx. 1:2000, approx. 1:200~approx. 1:1600, approx. 1:200~approx. 1:1200, approx. 1:200~approx. 1:800, approx. 1:200~approx. 1:400, approx. 1:280~approx. 1:40000, approx. 1:280~approx. 1:3000 0, approximately 1:280~1:20000, approximately 1:280~1:10000, approximately 1:280~1:5000, approximately 1:280~1:3000, approximately 1:280~1:2400, approximately 1:280~1:2000, approximately 1:280~1:1700, approximately 1:280~1:1600, approximately 1:280~approx. 1:1200, approx. 1:280~approx. 1:1100, approx. 1:280~approx. 1:800, approx. 1:280~approx. 1:560, approx. 1:280~approx. 1:400, approx. 1:400~approx. 1:40000, approx. 1:400~approx. 1:30000, approx. 1:400~approx. 1:20000, approx. 1:40 0 to approximately 1:8000, approximately 1:400 to approximately 1:4000, approximately 1:400 to approximately 1:3000, approximately 1:400 to approximately 1:2400, approximately 1:400 to approximately 1:2000, approximately 1:400 to approximately 1:1600, approximately 1:400 to approximately 1:1200, approximately 1:400 to approximately 1:800, approximately 1:560 to approximately 1:4 0000, approx. 1:560~1:30000, approx. 1:560~1:20000, approx. 1:560~1:8000, approx. 1:560~1:4000, approx. 1:560~1:3000, approx. 1:560~1:2400, approx. 1:560~1:2000, approx. 1:560~1:170 0, approximately 1:560~1:1600, approximately 1:560~1:1200, approximately 1:560~1:1100, approximately 1:560~1:800, approximately 1:800~1:40000, approximately 1:800~1:30000, approximately 1:800~1:20000, approximately 1:800~1:10000,Approximately 1:800 to 1:5000, approximately 1:800 to 1:3000, approximately 1:800 to 1:2400, approximately 1:800 to 1:2000, approximately 1:800 to 1:1600, approximately 1:800 to 1:1200, approximately 1:1100 to 1:40000, approximately 1:1100 to 1:30000, Approximately 1:1100~1:20000, Approximately 1:1100~1:10000, Approximately 1:1100~1:5000, Approximately 1:1100~1:4000, Approximately 1:1100~1:3000, Approximately 1:1100~1:2400, Approximately 1:1100~1:2000, Approximately 1:1100~1: 1700, approximately 1:1100~1:1600, approximately 1:1200~1:40000, approximately 1:1200~1:30000, approximately 1:1200~1:20000, approximately 1:1200~1:10000, approximately 1:1200~1:5000, approximately 1:1200~1:4000, approximately 1: 1200~approx. 1:3000, approx. 1:1200~approx. 1:2400, approx. 1:1200~approx. 1:2000, approx. 1:1200~approx. 1:1600, approx. 1:1400~approx. 1:1800, approx. 1:1600~approx. 1:40000, approx. 1:1600~approx. 1:30000, approx. 1:1600~approx. 1:200 00, approx. 1:1600~approx. 1:10000, approx. 1:1600~approx. 1:5000, approx. 1:1600~approx. 1:3000, approx. 1:1600~approx. 1:2400, approx. 1:1600~approx. 1:2000, approx. 1:1700~approx. 1:40000, approx. 1:1700~approx. 1:30000, approx. 1:170 0 to approximately 1:20000, approximately 1:1700 to approximately 1:10000, approximately 1:1700 to approximately 1:5000, approximately 1:1700 to approximately 1:3000, approximately 1:1700 to approximately 1:2400, approximately 1:1700 to approximately 1:2000, approximately 1:2000 to approximately 1:40000, approximately 1:2000 to approximately 1:30000 , approximately 1:2000~approximately 1:20000, approximately 1:2000~approximately 1:10000, approximately 1:2000~approximately 1:5000, approximately 1:2000~approximately 1:4000, approximately 1:2000~approximately 1:3000, approximately 1:2000~approximately 1:2400, approximately 1:2400~approximately 1:40000, approximately 1:2400~ Approximately 1:30000, approximately 1:2400~approximately 1:20000, approximately 1:2400~approximately 1:10000, approximately 1:2400~approximately 1:5000, approximately 1:2400~approximately 1:4000, approximately 1:2400~approximately 1:3000, approximately 1:3000~approximately 1:40000, approximately 1:3000~approximately 1:30000,It has an OT / Mg(m) ratio of approximately 1:3000 to 1:20000, approximately 1:3000 to 1:10000, approximately 1:3000 to 1:4000, approximately 1:4000 to 1:40000, approximately 1:4000 to 1:30000, approximately 1:4000 to 1:20000, approximately 1:4000 to 1:10000, approximately 1:8000 to 1:40000, approximately 1:8000 to 1:30000, approximately 1:8000 to 1:20000, or approximately 1:10000 to 1:40000. In one embodiment, the oxytocin peptide is human oxytocin consisting of Cys-Tyr-Ile-Gln-Asn-Cys-Pro-Leu-Gly (SEQ ID NO: 1).

[0052] In some embodiments, a magnesium-containing oxytocin peptide formulation or composition comprising an oxytocin peptide and a magnesium ion further comprises one or more pharmaceutically acceptable carriers (and thus constituting a pharmaceutical composition) and optionally other components (e.g., excipients, vehicles, emulsifiers, stabilizers, preservatives, buffers, and / or other additives that can enhance stability, delivery, absorption, half-life, efficacy, pharmacokinetics and / or pharmacodynamics, reduce adverse side effects, or provide other advantages to pharmaceutical use). Exemplary excipients include solubilizers, surfactants, and chelating agents. For example, the formulation may contain methyl-β-cyclodextrin (Me-β-CD), disodium edetate, arginine, sorbitol, NaCl, sodium methylparaben (MP), sodium propylparaben (PP), chlorobutanol (CB), benzyl alcohol, zinc chloride, ethyl alcohol, didecanoyl L-α-phosphatidylcholine (DDPC), polysorbate, lactose, citrate, tartrate, acetate and / or phosphate.

[0053] Examples of liquid carriers include, but are not limited to, water, saline solution, dextrose aqueous solution, and glycol, especially when the solution is isotonic. The carrier can also be selected from a variety of oils, including petroleum, animal, plant, or synthetic oils (e.g., peanut oil, olive oil, soybean oil, mineral oil, sesame oil, etc.). Suitable pharmaceutical excipients include, but are not limited to, starch, cellulose, talc, glucose, lactose, sucrose, gelatin, malt, rice, grain flour, chalk, silica gel, magnesium stearate, sodium stearate, glycerol monostearate, sodium chloride, skim milk powder, glycerol, propylene glycol, water, and ethanol. The composition may be subjected to conventional pharmaceutical processes such as sterilization and may contain conventional pharmaceutical additives (e.g., preservatives, stabilizers, reducing agents, antioxidants, chelating agents, wetting agents, emulsifiers, dispersants, gelling agents, salts for adjusting osmotic pressure, buffering agents, etc.). The liquid carrier may be hypotonic or isotonic with respect to body fluids and may have a pH in the range of 3.5 to 8.5. The use of additives in the preparation of peptide and / or protein-based compositions, particularly pharmaceutical compositions, is well known in the art. In some embodiments, the composition has a pH of about 2 to about 7. In some embodiments, the composition has a pH of about 4 to about 7. In preferred embodiments, the pH of the formulation / composition is about 4.5.

[0054] In some embodiments, a magnesium-containing oxytocin peptide formulation or composition may further comprise one or more mucosal delivery enhancers selected from (A) to (K): (A) solubilizers; (B) charge modifiers; (C) pH adjusters; (D) enzyme inhibitors; (E) mucus-clearing agents; (F) ciliostatic agents; (G) membrane permeability enhancers; (H) modifiers of the physiological function of epithelial junctions (e.g., nitric oxide (NO) stimulants, chitosan and chitosan derivatives); (I) vasodilators; (J) selective transport enhancers; and (K) stabilizing delivery vehicles, carriers, support or complex-forming species that effectively combine with, associate with, contain, encapsulate or bind to oxytocin peptides to stabilize the active agent for enhancing mucosal delivery. The membrane permeability enhancers in group (G) may be (i) surfactants, (ii) bile salts, (iii) phospholipid or fatty acid additives, mixed micelles, liposomes or carriers, (iv) alcohols, (v) enamines, (iv) NO donor compounds, (vii) long-chain amphiphilic molecules, (viii) small molecule hydrophobic permeability enhancers; (ix) sodium or salicylic acid derivatives; (x) glycerol esters of acetoacetic acid, (xi) cyclodextrin or beta-cyclodextrin derivatives, (xii) medium-chain fatty acids, (xiii) chelating agents, (xiv) amino acids or their salts, (xv) N-acetylamino acids or their salts, (xvi) enzymes that degrade selected membrane components, (xvii) inhibitors of fatty acid synthesis, (xviii) inhibitors of cholesterol synthesis; or any combination of (xiv)(i) to (xviii) membrane permeability enhancers. In various embodiments of the present invention, the oxytocin peptide may be combined with one, two, three, four or more of the mucosal delivery enhancers listed in (A) to (K). These mucosal delivery enhancers may be mixed with the oxytocin peptide alone or together, or combined with them in a formulation vehicle or delivery vehicle that is otherwise pharmaceutically acceptable.The magnesium-containing oxytocin peptide preparations or compositions described herein may increase the bioavailability of the oxytocin peptide after being delivered to the mucosal surface of a mammalian subject (e.g., the mucosal surface of the nasal cavity).

[0055] The list of carriers and excipients discussed herein is by no means exhaustive, and those skilled in the art can select carriers and excipients from the GRAS (generally regarded as safe) list of chemicals permitted in pharmaceuticals, as well as chemicals currently permitted by the U.S. Food and Drug Administration in topical and parenteral formulations, and chemicals that may be permitted in the future (see also Wang et al. (1980) J.Parent. Drug Assn., 34:452-462; Wang et al. (1988) J.Parent. Sci. and Tech., 42:S4-S26).

[0056] In some embodiments, a magnesium-containing oxytocin peptide preparation or composition comprising an oxytocin peptide and magnesium ions (where the oxytocin peptide and magnesium ions are present in amounts that produce a synergistically enhanced effect when used in the treatment of autism spectrum disorder) further comprises one or more solvents or excipients selected from the group consisting of chlorobutanol, benzalkonium, methyl 4-hydroxybenzoate, propyl 4-hydroxybenzoate, acetic acid, citric acid, glycerol, sodium chloride, monohydrogen phosphate, sorbitol, and water. In some embodiments, the magnesium-containing oxytocin peptide preparation or composition further comprises chlorobutanol, acetic acid, and water.

[0057] In some embodiments, a magnesium-containing oxytocin peptide formulation or composition comprising an oxytocin peptide and magnesium ions further comprises a chitosan-containing excipient (e.g., ChiSys®, http: / / www.archimedespharma.com / productArchiDevChiSys.html). In some embodiments, the magnesium-containing oxytocin peptide formulation or composition further comprises about 1% of the chitosan-containing excipient. In some embodiments, chitosan glutamate may be preferred for nasal delivery due to its excellent absorption-enhancing ability. In some embodiments, chitosan copolymer nanoparticles (e.g., nanoparticles comprising chitosan glutamate and a negatively charged polymer (e.g., pentasodium tripolyphosphate)) may be used. Thiolated chitosan (e.g., chitosan covalently modified with 2-iminothiolane) has been used in microparticles containing insulin and reduced glutathione and may also be useful as an excipient in the magnesium-containing oxytocin peptide formulations or compositions described herein.

[0058] In some embodiments, a magnesium-containing oxytocin peptide formulation or composition comprising an oxytocin peptide and magnesium ions further comprises one or more gelling agents to enhance the nasal absorption of the oxytocin peptide, so that the oxytocin peptide formulation forms a gel in the nasal cavity. Useful gelling systems in the formulations and methods described herein may include any known gelling systems, e.g., chemically reactive pectin-based gelling systems (e.g., PecSys®, Archimedes Pharma) and thermally reactive polymer gelling systems (e.g., Pluronic® F127, BASF). PecSys® is a low-viscosity pectin-based aqueous solution delivered as a fine mist, where each droplet gels upon contact with calcium ions in the nasal mucosa. Other low-methoxypectins may also be used, for example, at concentrations of about 1%. Pluronic® F127 comprises an ethylene oxide / propylene oxide block copolymer. The gelling temperature varies depending on the ratio of the components and the amount of copolymer used in the final formulation. Gelation in the human nasal cavity has been demonstrated, for example, for vitamin B12 gel supplements (EnerB, Nature's Bounty, NY) and for gelled sumatriptans containing approximately 18–20% wt / vol Pluronic® F127 and 0.3% wt / vol Carbopol (anionic bioadhesive polymer C934P). The monomer ratios and concentrations can be adjusted for the intended oxytocin formulation to ensure gelation at 25–37°C, which is around the standard temperature of 34°C in the nasal cavity. If the gelation temperature is below 25°C, the formulation may gel at room temperature; if the gelation temperature is above 37°C, the formulation will not gel completely upon contact with the nasal mucosa. In some embodiments, the magnesium-containing oxytocin peptide formulation or composition may further contain a mucosal adhesive substance such as Carbopol. The gelation temperature can be further reduced by adding a mucosal adhesive, for example, up to 0.5% of Carbopol.

[0059] In some embodiments, magnesium-containing oxytocin peptide formulations or compositions comprising oxytocin peptides and magnesium ions further comprise a nonionic surfactant (e.g., polysorbate-80) and one or more surfactants such as buffers, stabilizers, or isotonic agents (tonicifiers). In some embodiments, magnesium-containing oxytocin peptide formulations or compositions further comprise a propellant. The pH of the nasal spray solution is, if necessary, about pH 3.0 to 8.5, but if desired, the pH is adjusted to optimize the delivery of charged polymer species (e.g., therapeutic proteins or peptides) in a substantially unionized state. The pharmaceutical solvent used may also be a slightly acidic aqueous buffer (pH 3 to 6). Suitable buffers for use in these compositions are as described above or otherwise known in the art. Other components, including preservatives, surfactants, dispersants, or gases, may be added to enhance or maintain chemical stability. Suitable preservatives include, but are not limited to, phenol, methylparaben, parabens, m-cresol, thiomersal, and benzalkonium chloride. Suitable surfactants include, but are not limited to, oleic acid, sorbitan trioleate, polysorbate, lecithin, phosphotidylcholine, and various long-chain diglycerides and phospholipids. Suitable dispersants include, but are not limited to, ethylenediaminetetraacetic acid (EDTA). Suitable gases include, but are not limited to, nitrogen, helium, chlorofluorocarbons (CFCs), hydrofluorocarbons (HFCs), carbon dioxide, and air. Suitable stabilizers and tonicizing agents include sugars and other polyols, amino acids, and organic and inorganic salts. In some embodiments, the magnesium-containing oxytocin peptide formulation or composition further comprises citrate, succinate, or pyrophosphate.

[0060] In some embodiments, the magnesium-containing oxytocin peptide preparation or composition, comprising an oxytocin peptide and magnesium ions, further comprises an agent (e.g., IL-6) capable of upregulating oxytocin receptor expression.

[0061] To further enhance mucosal delivery of oxytocin peptides, enzyme inhibitors, particularly protease inhibitors, may be included in the above formulations. Examples of protease inhibitors include, but are not limited to, antipain, arphamenine A and B, benzamidine HCl, AEBSF, CA-074, calpain inhibitors I and II, calpeptin, pepstatin A, actinonine, amastatin, bestatin, boroleucine, captopril, chloroacetyl-HOLeu-Ala-Gly-NH2, DAPT, diprotin A and B, ebelactone A and B, foroxymithine, leupeptin, phosphoramidone, aprotinin, puromycin, BBI, soy trypsin inhibitors, phenylmethylsulfonyl fluoride, E-64, chymostatin, 1,10-phenanthroline, EDTA, and EGTA. Other enzyme inhibitors such as bacitracin may also be included in the above formulations.

[0062] Absorption enhancers may be included in the formulation to enhance the delivery of oxytocin peptide and magnesium ions into or beyond the mucosal surface and / or absorption. These enhancers may increase the release or solubility of the composition (e.g., from the formulation delivery vehicle), diffusion rate, permeation capacity and timing, uptake, residence time, stability, effective half-life, peak or sustained concentration levels, clearance, and other desired mucosal delivery properties (e.g., when measured at the delivery site). Therefore, enhanced mucosal delivery may occur through any of the following mechanisms, for example, by increasing the diffusion, transport, persistence or stability of oxytocin peptides; increasing membrane fluidity; regulating the availability or action of calcium and other ions that control intracellular or paracellular permeability; solubilizing mucosal components (e.g., lipids); altering sulfhydryl levels of non-proteins and proteins in mucosal tissue; increasing water flow across the mucosal surface; regulating the physiological function of epithelial junctions; decreasing the viscosity of the mucus covering the mucosal epithelium; decreasing the clearance rate of mucociliary tissue; and other mechanisms.

[0063] Compounds that enhance mucosal absorption may include, but are not limited to, surfactants, bile salts, dihydrofusidetes, bioadhesives / mucosal adhesion substances, phospholipid additives, mixed micelles, liposomes or carriers, alcohols, enamines, cationic polymers, NO donor compounds, long-chain amphiphilic molecules, small molecule hydrophobic permeability enhancers; sodium derivatives or salicylic acid derivatives, glycerol esters of acetoacetate, cyclodextrins or beta-cyclodextrin derivatives, medium-chain fatty acids, chelating agents, amino acids or their salts, N-acetyl amino acids or their salts, mucolytics, enzymes specifically targeted to selected membrane components, inhibitors of fatty acid synthesis, and inhibitors of cholesterol synthesis.

[0064] All peptides described and / or intended herein can be prepared by chemical synthesis using automated or manual solid-phase synthesis techniques widely known in the art. These peptides can also be prepared using molecular recombination methods known in the art.

[0065] (Delivery system) Magnesium-containing oxytocin peptide preparations or compositions may be adapted for craniofacial mucosal administration (e.g., nasal, buccal, sublingual, or ocular administration). In some embodiments, the composition may further constitute a device for mucosal delivery. In some embodiments, the composition is adapted for buccal and / or sublingual mucosal delivery, and the composition may further constitute a device for buccal and / or sublingual mucosal administration (e.g., unit dose containers, pump sprays, droppers, squeeze bottles, preservative-free airless sprays, nebulizers, dose inhalers, and pressurized dose inhalers). In some embodiments, the composition is adapted for ocular delivery, and the composition may further constitute a device for conjunctival administration (e.g., droppers or squeeze bottles). In some embodiments, the composition is adapted for intranasal administration, and the composition may further constitute a device for intranasal administration (e.g., droppers, pump sprays, squeeze bottles, preservative-free airless sprays, or nasal pump devices, e.g., nasal pump devices with a storage bottle attached to an aerozoizer).

[0066] Intranasal drug delivery has been a research and development topic for many years, but carrier systems that effectively deliver substances have only been devised within the last 10 years (Sayani and Chien, Critical Reviews in Therapeutic Drug Carrier Systems 1996, 13:85-184). Intranasal delivery offers several beneficial features, including relatively high bioavailability, rapid absorption kinetics, and avoidance of the first-pass effect in the liver. In some embodiments, intranasal administration may enable delivery of oxytocin peptides into the nasal cavity, and in other embodiments, intranasal administration may enable targeted delivery to cranial nerves in the nose and / or brain. While we do not wish to be bound by any particular theory, intranasal administration of oxytocin peptides may target either or both of the olfactory nervous system or the trigeminal nervous system. Oxytocin peptides may be delivered intranasally in any applicable form, including, but not limited to, liquid formulations, solid formulations (e.g., dry powder formulations), gel formulations, or emulsion formulations.

[0067] In embodiments in which a combination of oxytocin and magnesium ions is administered intranasally, the composition may be prepared as a liquid aerosol formulation combined with a dispersant and / or a physiologically acceptable diluent. Alternatively, a dry powder aerosol formulation may be intended, which may comprise the subject compound in a finely pulverized solid form and a dispersant that enables rapid dispersion of the dry powder particles. Whether a liquid aerosol formulation or a dry powder aerosol formulation, the formulation is aerosolized into small liquid or solid particles to ensure that the aerosolized dose reaches the mucosa of the nasal tract or the lungs. The term “aerosol particles” is used herein to describe suitable liquid or solid particles with a particle diameter small enough for distribution to the nasal mucosa or alveolar membranes (within the range of about 10 microns) or lungs (within the range of about 2 to 5 microns). Other considerations include the structure of the delivery device, further components in the formulation, and the properties of the particles. These embodiments of intranasal or pulmonary administration of drugs are well known in the art, and the formulation operations, aerosolization means and the structure of the delivery device are within the normal level of the art.

[0068] In some embodiments, a magnesium-containing oxytocin peptide formulation or composition useful in the methods described herein, and in which the oxytocin peptide and magnesium ions are present in amounts that produce a synergistic or enhanced effect when used in the treatment of autism spectrum disorder, is administered using a device for intranasal delivery. The device may be any device suitable for intranasal administration of a magnesium-containing oxytocin peptide formulation. In some embodiments, the device is suitable for delivery of the oxytocin peptide and magnesium ions to a specific area within the nasal cavity. In some embodiments, the device is suitable for delivery of the oxytocin peptide and magnesium ions to the lower two-thirds of the nasal cavity. In some embodiments, the device is suitable for delivery of the oxytocin peptide and magnesium ions to the upper one-third of the nasal cavity. In some embodiments, the device is suitable for delivery of the oxytocin peptide to the entire nasal tract.

[0069] In some embodiments, the device for intranasal delivery is a nasal pump device. In some embodiments, the nasal pump device comprises a storage bottle attached to a pump actuator. In some embodiments, the pump actuator meters and dispenses a specific volume (e.g., about 5 to about 1000 μL, preferably about 50 to about 150 μL, more preferably about 50 μL or about 100 μL) to be delivered in a specific droplet size distribution. In some embodiments, the nasal pump device comprises a storage bottle attached to an aerozoizer, e.g., the Equadel pump sold by Aptar Pharma. In some embodiments, the device for nasal administration functions regardless of the pressure applied to the pump once a threshold is reached. In some embodiments, the device for intranasal administration may be a mucosal spray device that can be attached to a syringe (e.g., LMA® MAD NASAL TMFor administration in large mammals, the nasal pump device may include a storage bottle attached to a pump actuator that metered and dispensed a larger volume (e.g., about 100 μL to about 600 μL or more) to deliver.

[0070] In some embodiments, intranasal delivery devices are designed to deliver multiple doses of drug formulations. For example, a nasal pump device may include a storage bottle attached to a pump actuator, the storage bottle holding multiple doses of liquid formulation, and the pump actuator metering and dispensing to deliver a specific volume which is part of the liquid formulation held in the storage bottle. In some embodiments, the pump actuator metering and dispensing to deliver approximately 50 μL of liquid formulation per spray. The nasal pump device may include a filter to prevent backflow in order to reduce the entry of contaminants (e.g., bacteria) into the storage bottle. In some embodiments, the nasal pump device includes a metal-free channel (e.g., a plastic channel) for delivering the liquid formulation. In some embodiments, the pump device uses a plastic material that is stable to gamma rays (used to sterilize the nasal device). In some embodiments, the intranasal delivery device is equipped with a multiple-dose dosing pump with a microbial filter and an automatic blocking mechanism on the pump actuator, for example, the spray device described in U.S. Patent No. 5,988,449.

[0071] In some embodiments, the intranasal delivery device is a breath-actuated nasal delivery device (e.g., the devices described in U.S. Patents 7,784,460 and 7,854,227). Such devices can improve delivery to deeper target sites within the nasal cavity. In some embodiments, a standard metered-dose spray device is incorporated into a housing in which the patient breathes into a mouthpiece to activate the device. In some embodiments, the device comprises a conical, sealed nosepiece and mouthpiece incorporating a conventional mechanical spray pump (e.g., the Equadel pump sold by Aptar Pharma), a chargeable spring, and an exhalation-actuated mechanism. The system can be used for single-dose or multi-dose delivery. An example of such a liquid delivery device is the OptiMist® device sold by OptiNose. When using this device, the nosepiece is inserted into the nostril and breath is exhaled towards the mouthpiece. This causes the soft palate to close, pressurizing the nostrils and opening a passage behind the nasal septum, allowing air to flow out through the other nostril (two-way flow). Because the device is activated by exhalation, small particles cannot enter the lungs. By modifying the flow rate and particle size, it becomes possible to target specific nasal regions.

[0072] In some embodiments, the intranasal delivery device is a unit dose metering and dispensing spray device suitable for a single dose of a magnesium-containing oxytocin peptide preparation or composition. In some embodiments, the intranasal delivery device is a multi-dose metering and dispensing spray pump device suitable for repeated doses of oxytocin peptides.

[0073] Droplet size, plume volume, and flow rate can be modified to target specific nasal regions. Liquid sprays can offer droplet sizes of 5–50 microns to target the olfactory epithelium and / or airway epithelium. Larger droplets primarily fall into the nasopharynx and are swallowed, while smaller droplets target lung tissue. The Mass Median Equivalent Aerodynamic Diameter (MMAD) is used to determine the droplet size. The pH of nasal sprays is optimized to deliver charged peptides in a nearly unionized state. The nose typically tolerates solutions with a pH of approximately 3–8. The nasal mucosa can typically absorb a volume of approximately 100 μL until saturation occurs and the liquid begins to drip out of the nose. Therefore, the plume volume can be up to 100 μL (and including 100 μL). When used in large mammals, the plume volume may be up to 150 μL (and including 150 μL) or greater (e.g., 600 μL or greater). For use in infants and children, or for veterinary use in small animals (e.g., rodents, cats), smaller plume volumes (5–50 μL) may be used.

[0074] In some embodiments, the intranasal delivery device is ergonomically designed to facilitate patient adherence to medication (e.g., a pump device with a side-driven trigger mechanism). In some embodiments, the intranasal delivery device includes a metered-feed spray pump that operates as a closed system to prevent contamination by airborne microorganisms by preventing air from entering the pump device. In some embodiments, the intranasal delivery device includes a metered-feed spray pump that operates with a filter. Passing air is drawn in through a filter assembled in the pump, thereby keeping airborne microorganisms outside the pump device. In some embodiments, the intranasal delivery device with a nasal pump device may further include a microelectronic device that can facilitate data transmission and monitoring of the procedure.

[0075] In some embodiments, a magnesium-containing oxytocin peptide formulation or composition comprises an oxytocin peptide and a magnesium ion, wherein the oxytocin peptide and magnesium ion are contained in one of the intranasal delivery devices described herein, and the concentrations of the oxytocin peptide and magnesium ion are within any of the concentration ranges described herein, as each combination of device and concentration and all combinations are described individually.

[0076] (method) The term “autism spectrum disorder (ASD)” or “autism” refers to a group of complex disorders of brain development. These disorders are characterized to varying degrees by difficulties in social interaction, verbal and nonverbal communication, and repetitive behaviors. In May 2013, the Diagnostic and Statistical Manual of Mental Disorders, Fifth Edition (DSM-5) was published, and all autism disorders were integrated into a single comprehensive diagnosis, ASD. Previously, they were recognized as separate subtypes (including autistic disorder, childhood disintegrative disorder, pervasive developmental disorder not otherwise specified (PDD-NOS), and Asperger's syndrome). See http: / / www.autismspeaks.org / what-autism. Those skilled in the art will recognize that the symptoms of autism spectrum disorder overlap considerably with many other mental disorders. Examples of disorders that exhibit symptoms similar to those shown in autism spectrum disorder include, but are not limited to, social anxiety disorder, obsessive-compulsive disorder, social (pragmatic) communication disorder, and neurodevelopmental disorders (including, but not limited to, attention deficit hyperactivity disorder, Prader-Willi syndrome, Timothy syndrome, fragile X syndrome, Rett syndrome, and Williams syndrome).

[0077] The DSM-5 provides diagnostic criteria for ASD that include: (A) Persistent deficits in social communication and social interaction across multiple contexts, present or historically, as illustrated by: (1) Deficiencies in social-emotional reciprocity (e.g., from abnormal social approaches and inability to engage in normal conversation; to reduced sharing of interests, emotions, or attachments; to inability to initiate or respond to social interactions); (2) Deficiencies in nonverbal communication behaviors used in social interaction (e.g., from poor integration of verbal and nonverbal communication; to eye contact and body language). (1) Abnormalities in communication or impairments in understanding and using gestures; ranging from a complete lack of facial and nonverbal communication; and (2) impairments in developing, maintaining and understanding relationships (ranging from, for example, difficulty in adapting behavior to different social contexts; from difficulty in sharing creative play or making friends; to a lack of interest in peers); and (3) restriction of repetitive patterns of behavior, interests, or activities, as manifested, currently or in the past, by at least two of the following examples: (1) stereotypic or repetitive movements, use of objects, or speech (for example, simple stereotypic movements (motor (1) Stereotypy, arranging toys or turning objects upside down, echolalia, distinctive speech patterns; (2) Obsession with the same thing, stubborn adherence to a set procedure, or ritualized patterns or verbal / nonverbal behaviors (e.g., extreme distress at minor changes, difficulties with transitions, rigid thought patterns, greeting rituals, needing to take the same route or eat the same food every day); (3) Extremely localized and fixated interests in intensity or at the center of interest (e.g., strong attachment or obsession with unusual objects, excessively localized or fixated interests);(4) hypersensitivity or hyposensitivity to sensory input, or an unusual interest in the sensory aspects of the environment (e.g., appearing indifferent to pain / temperature, adverse reaction to certain sounds or textures, excessive smelling or touching of objects, or intensely watching light or movement). See http: / / www.autismspeaks.org / what-autism / diagnosis / dsm-5-diagnostic-criteria.

[0078] Autism spectrum disorder (ASD) is characterized by difficulties in social interaction, communication difficulties, and a tendency to engage in repetitive behaviors. However, symptoms and their severity vary widely across these three core areas. ASD may be associated with intellectual disability, motor coordination difficulties, and attention and physical health problems (e.g., sleep and gastrointestinal disorders). ASD may also be associated with psychiatric symptoms, including anxiety and depression. See, for example, Kim et al., Autism 2000, 4(2):117-132.

[0079] Oxytocin has been known to treat many conditions in autism spectrum disorder, including anxiety and social and communication deficits. However, the effects of oxytocin on social and communication deficits in autism spectrum disorder have been observed to vary widely among patients. Variations in receptor availability and affinity for oxytocin are thought to be the cause of these variations in effects. Clinical efforts to treat ASD with commercially available oxytocin formulations (e.g., Syntocinon®) have been hampered by lack of efficacy and poor tolerability. Due to the low potency and high volume of currently available oxytocin formulations, the amount of drug absorbed when administered by nasal spray is insufficient in terms of efficacy. The present invention provides a method for administering oxytocin peptides in a more potent formulation and a lower volume, so that an effective amount of the formulation can be delivered using a nasal device for treating autism spectrum disorder, a disorder exhibiting one or more symptoms associated with autism spectrum disorder, or social and communication deficits.

[0080] In one embodiment, a method is provided for treating autism spectrum disorder, a disorder exhibiting one or more symptoms associated with autism spectrum disorder, social and communication deficits, or anxiety, the method comprising the step of administering an effective amount of oxytocin peptide and magnesium ions to a subject in need, wherein the effective amount is delivered via intranasal administration in a volume readily absorbed in the nasal cavity. In some embodiments, the volume that allows for readily absorbed oxytocin peptide and magnesium ions in the nasal cavity is about 5 μL to about 1000 μL. In some embodiments, the molar ratio of the amount of oxytocin peptide to the amount of magnesium or magnesium ions is about 1:175, about 1:280, about 1:560, about 1:1100, about 1:1700, or about 1:2000 (including any range between these ratios).

[0081] In one embodiment, a method is provided comprising the step of administering an effective amount of oxytocin peptide and magnesium ions to a subject in need, wherein the effective amount is delivered via intranasal administration in a volume of about 5 μL to about 1000 μL. In some embodiments, the molar ratio of the amount of oxytocin peptide to the amount of magnesium or magnesium ions is about 1:175, about 1:280, about 1:560, about 1:1100, about 1:1700, or about 1:2000 (including any range between these ratios). In some embodiments, the method is for treating one or more symptoms associated with autism spectrum disorder. In some embodiments, the method is for treating disorders exhibiting one or more symptoms associated with autism spectrum disorder. In some embodiments, the method is for reducing social and communication deficits. In some embodiments, the method is for treating or reducing anxiety.

[0082] Magnesium is involved in many aspects of life and health (e.g., energy production, oxygen uptake, central nervous system function, electrolyte balance, glucose metabolism, and muscle activity). Magnesium has also been found to be clinically effective in reducing social and communication deficits in children with autism spectrum disorder. See Mousain-Bosc et al., Magnes. Res. 2006, 19(1):53-62. The co-administration of oxytocin and magnesium ions of this invention produces synergistic or enhanced improvements in social behavior and reductions in anxiety compared to oxytocin administration alone. The mechanisms underlying these effects are unclear, but they appear to involve either non-competitive blockade of the N-methyl D-aspartate (NMDA) neurotransmitter receptor, or increased affinity for oxytocin receptor activity as an allosteric modulator, or both.

[0083] In some embodiments, a method is provided for treating autism spectrum disorder, the method comprising the step of administering an effective dose of oxytocin peptide and magnesium ions to a subject in need, wherein the simultaneous administration of the oxytocin peptide and magnesium ions results in a synergistic or enhanced effect. In some embodiments, a method is provided for mitigating or reducing one or more symptoms associated with autism spectrum disorder, the method comprising the step of administering an effective dose of oxytocin peptide and magnesium ions to a subject in need, wherein the simultaneous administration of the oxytocin peptide and magnesium ions results in a synergistic or enhanced effect. In some embodiments, a method is provided for treating a disorder exhibiting one or more symptoms associated with autism spectrum disorder, the method comprising the step of administering an effective dose of oxytocin peptide and magnesium ions to a subject in need, wherein the simultaneous administration of the oxytocin peptide and magnesium ions results in a synergistic or enhanced effect. In some embodiments, oxytocin peptides and magnesium ions are administered in doses that produce an overall effect on reducing or alleviating the symptoms that is greater than the sum of the effects of equal doses of oxytocin peptides and magnesium salts administered individually. In some embodiments, oxytocin peptides and magnesium ions are administered in doses that produce an effect that is faster onset and / or lasts longer than that that occurs after administration of the individual drugs used alone in equal amounts. Examples of symptoms associated with autism spectrum disorder include, but are not limited to, persistent deficits in social communication and social interaction, social anxiety, and restricted behaviors, interests and activities. Other behaviors and characteristics observed in individuals with autism spectrum disorder also include avoidance of physical contact, generalized anxiety, monotony or inability to regulate vocal volume, inability to develop peer relationships, lack of shared enjoyment and interests, and lack of social or emotional reciprocity.Examples of disorders that exhibit symptoms similar to those shown in autism spectrum disorder include, but are not limited to, social anxiety disorder, obsessive-compulsive disorder, social (pragmatic) communication disorder, and neurodevelopmental disorders (including, but not limited to, attention deficit hyperactivity disorder, Prader-Willi syndrome, Timothy syndrome, fragile X syndrome, Rett syndrome, and Williams syndrome).

[0084] Prader-Willi syndrome is a complex genetic condition that affects many parts of the body and is caused by a loss of function of a gene in a specific region of chromosome 15. Individuals with Prader-Willi syndrome often have mild to moderate intellectual impairment and learning difficulties, and many exhibit behavioral problems, including mood swings, stubbornness, manipulative behavior, and obsessive-compulsive behaviors, such as scratching their skin. Other symptoms often observed in individuals with Prader-Willi syndrome include persistent deficits in social communication and social interaction, anxiety and irritability, and sleep problems.

[0085] In some embodiments, a method for treating Prader-Willi syndrome is provided, the method comprising the step of administering an effective dose of oxytocin peptide and magnesium ions to a subject in need thereof. In some embodiments, the simultaneous administration of the oxytocin peptide and magnesium ions results in a synergistic or enhanced effect. In some embodiments, the oxytocin peptide and magnesium ions are administered in a dose that produces a larger, overall effect than summing the effects of equal doses of the oxytocin peptide and magnesium salt administered individually. In some embodiments, the oxytocin peptide and magnesium ions are administered in a dose that produces an effect that is faster onset and / or lasts longer than that that occurs after administration of the individual drugs used alone in equal amounts. In one embodiment, the present invention provides a method for treating Prader-Willi syndrome, the method comprising the step of administering an effective dose of oxytocin peptide and magnesium ions to a subject in need thereof, the effective dose of oxytocin peptide and magnesium ions being administered intranasally in a liquid formulation, the volume of the liquid formulation administered being about 5 μL to about 1000 μL. In some embodiments, the molar ratio of the amount of oxytocin peptide to the amount of magnesium or magnesium ions is about 1:175, about 1:280, about 1:560, about 1:1100, about 1:1700, or about 1:2000 (including any range between these ratios).

[0086] In some embodiments, methods are provided for alleviating or reducing one or more symptoms associated with Prader-Willi syndrome, the methods comprising the step of administering an effective dose of oxytocin peptide and magnesium ions to a subject in need. Examples of symptoms associated with Prader-Willi syndrome include, but are not limited to, persistent deficits in social communication and social interaction, anxiety and irritability, and sleep problems. In some embodiments, the co-administration of the oxytocin peptide and magnesium ions results in a synergistic or enhanced effect. In some embodiments, the oxytocin peptide and magnesium ions are administered in doses that produce an overall effect on alleviating or reducing symptoms that is greater than the sum of the effects of equal doses of the oxytocin peptide and magnesium salt administered individually. In some embodiments, the oxytocin peptide and magnesium ions are administered in doses that produce an effect that is faster onset and / or lasts longer than that that occurs after administration of the individual drugs used alone in equal amounts. Examples of symptoms associated with Prader-Willi syndrome include, but are not limited to, persistent deficits in social communication and social interaction, anxiety and irritability, and sleep problems. In one embodiment, the present invention provides a method for treating Prader-Willi syndrome, the method comprising the step of administering an effective dose of oxytocin peptide and magnesium ions to a subject in need thereof, wherein the effective dose of oxytocin peptide and magnesium ions is administered intranasally in a liquid formulation, the volume of which is about 5 μL to about 1000 μL. In some embodiments, the molar ratio of the amount of oxytocin peptide to the amount of magnesium or magnesium ions is about 1:175, about 1:280, about 1:560, about 1:1100, about 1:1700, or about 1:2000 (including any range between these ratios).

[0087] In some embodiments, methods are provided for treating anxiety associated with Prader-Willi syndrome, the methods comprising the step of administering an effective dose of oxytocin peptide and magnesium ions to a subject in need thereof. In some embodiments, the co-administration of the oxytocin peptide and magnesium ions results in a synergistic or enhanced effect. In some embodiments, the oxytocin peptide and magnesium ions are administered in doses that produce an overall effect on reducing or alleviating anxiety that is greater than the sum of the effects of equal doses of the oxytocin peptide and magnesium salt administered individually. In some embodiments, the oxytocin peptide and magnesium ions are administered in doses that produce an effect that has a faster onset and / or lasts longer than that that occurs after administration of the individual drugs used alone in equal amounts. In one embodiment, the present invention provides a method for treating anxiety associated with Prader-Willi syndrome, the method comprising the step of administering an effective dose of oxytocin peptide and magnesium ions to a subject in need, wherein the effective dose of oxytocin peptide and magnesium ions is administered intranasally in a liquid formulation, the volume of which is about 5 μL to about 1000 μL. In some embodiments, the molar ratio of the amount of oxytocin peptide to the amount of magnesium or magnesium ions is about 1:175, about 1:280, about 1:560, about 1:1100, about 1:1700, or about 1:2000 (including any range between these ratios).

[0088] In one embodiment, the present invention provides a method for treating deficiencies in sociality and communication, the method comprising administering an effective dose of oxytocin peptide and magnesium ions to a subject in need, wherein the simultaneous administration of the oxytocin peptide and magnesium ions results in a synergistic or enhanced effect. In one embodiment, the present invention provides a method for treating anxiety, the method comprising administering an effective dose of oxytocin peptide and magnesium ions to a subject in need, wherein the simultaneous administration of the oxytocin peptide and magnesium ions results in a synergistic or enhanced effect. In some embodiments, the oxytocin peptide and magnesium ions are administered in doses that produce an overall effect on reducing deficiencies in sociality and communication and / or anxiety, which is greater than the sum of the effects of equal doses of the oxytocin peptide and magnesium salt administered individually. In some embodiments, the molar ratio of oxytocin peptide to magnesium or magnesium ions is approximately 1:175, approximately 1:280, approximately 1:560, approximately 1:1100, approximately 1:1700, or approximately 1:2000 (including any range between these ratios). In some embodiments, deficiencies in sociality and communication include impaired communication skills and / or dysfunction in social interaction, lack of eye contact, and / or inability to form and / or maintain social relationships.

[0089] In some embodiments, methods are provided for treating anxiety associated with autism spectrum disorder, the methods comprising the step of administering an effective dose of oxytocin peptide and magnesium ions to a subject in need. In some embodiments, the co-administration of the oxytocin peptide and magnesium ions results in a synergistic or enhanced effect. In some embodiments, the oxytocin peptide and magnesium ions are administered in doses that produce an overall effect on reducing or alleviating anxiety that is greater than the sum of the effects of equal doses of the oxytocin peptide and magnesium salt administered individually. In some embodiments, the oxytocin peptide and magnesium ions are administered in doses that produce an effect that has a faster onset and / or lasts longer than that that occurs after administration of the individual drugs used alone in equal amounts. In some embodiments, the molar ratio of the amount of oxytocin peptide to the amount of magnesium or magnesium ions is about 1:175, about 1:280, about 1:560, about 1:1100, about 1:1700, or about 1:2000 (including any range between these ratios).

[0090] Oxytocin peptide and magnesium ions may be administered together or sequentially. In some embodiments, oxytocin peptide is administered simultaneously with magnesium ions in the same unit dose. In some embodiments, oxytocin peptide is administered simultaneously with magnesium ions but in separate unit doses or formulations. In some embodiments, oxytocin peptide and magnesium ions are administered sequentially. In some embodiments, magnesium ions are administered to the subject in a first dose, and then oxytocin peptide is administered to the subject in a second dose. In some of these embodiments, oxytocin peptide is administered approximately 10 minutes to approximately 2 hours after the administration of magnesium ions. In some of these embodiments, oxytocin peptide is administered approximately 10 minutes to approximately 2 hours, approximately 10 minutes to approximately 1 hour, approximately 10 minutes to approximately 30 minutes, approximately 20 minutes to approximately 2 hours, approximately 20 minutes to approximately 1 hour, approximately 30 minutes to approximately 2 hours, or approximately 30 minutes to approximately 1 hour after the administration of magnesium ions. In some of these embodiments, the oxytocin peptide is administered approximately 10 minutes, 15 minutes, 20 minutes, 30 minutes, 45 minutes, 60 minutes, 90 minutes, or 120 minutes after the administration of magnesium ions. In some of these embodiments, the oxytocin peptide is administered approximately 10 minutes, 15 minutes, 20 minutes, or 30 minutes after the administration of magnesium ions. In one embodiment, the oxytocin peptide is administered to the subject first, followed by the administration of magnesium ions. In some embodiments, the subject is human.

[0091] Interleukin-6 (IL-6) has been shown to induce increased oxytocin receptor expression in various tissues (e.g., Young et al., J. Neuroendocrinology, 1997; 9:859-65). Therefore, serum IL-6 levels can be used as a biomarker of the potential efficacy of oxytocin, for example, when magnesium is administered nasally.

[0092] In some embodiments, IL-6 is used as a biomarker of the effectiveness of oxytocin peptide administration in subjects and for subject selection for application of the methods described herein for treating autism spectrum disorder, disorders exhibiting one or more symptoms associated with autism spectrum disorder, social and communication deficits, or anxiety. In some embodiments, IL-6 is used for subject selection (e.g., human) for administration of oxytocin peptide (e.g., intranasal administration of oxytocin peptide in combination with magnesium ions).

[0093] In some embodiments, subjects are selected for treatment based on having high levels of IL-6. IL-6 levels may be higher than the control or reference. In some embodiments, IL-6 levels are high compared to the control or reference if they are significantly higher than the control or reference, as determined by appropriate statistical analysis. In some embodiments, IL-6 levels are high compared to the control or reference if they are at least one standard deviation higher than the control or reference. In some embodiments, the control is a value for IL-6 levels as determined in age-matched and sex-matched healthy subjects. In some embodiments, the reference is a reported value for IL-6 levels (e.g., a reported value for IL-6 in age-matched and sex-matched healthy subjects). In some embodiments, IL-6 levels are determined as the level of IL-6 in a sample derived from the subject (such as a tissue or liquid sample) (including, but not limited to, whole blood, serum, plasma, tears, etc.). The level of IL-6 in a sample may be determined by any method known in the art (e.g., by an immunoassay, e.g., an ELISA-based assay). See, for example, Yang, CJ., et al. Neuroscience 284: 290-296, 2015; Emanuele, E., et al. Neuroscience letters 471(3): 162-165, 2010; Ashwood, P., et al. Brain, behavior, and immunity 25(1): 40-45, 2011; and Malik, M., et al. Immunobiology 216(1): 80-85, 2011.

[0094] In some embodiments, methods for treating autism spectrum disorder, one or more symptomatic disorders associated with autism spectrum disorder, social and communication deficits, or anxiety include the steps of measuring the level of IL-6 (e.g., serum level of IL-6) in a subject, and administering an effective dose of oxytocin peptide and magnesium ions to a subject having high IL-6 levels.

[0095] In one embodiment, a method for treating autism spectrum disorder, one or more symptomatic disorders associated with autism spectrum disorder, social and communication deficits, or anxiety, comprising the step of administering an effective dose of oxytocin peptide and magnesium ions to a subject in need thereof, wherein the simultaneous administration of the oxytocin peptide and magnesium ions results in a synergistic or enhanced effect, further comprising the step of administering an effective dose of interleukin-6 (IL-6) to the subject. In some embodiments, the effective dose of IL-6 results in increased expression of oxytocin receptors (OTRs) in the subject.

[0096] In some embodiments, IL-6 is administered to a subject according to any of the methods described herein, and the oxytocin peptide and IL-6 may be administered simultaneously or sequentially. In some embodiments, the oxytocin peptide is administered simultaneously with IL-6 in the same unit dose. In some embodiments, the oxytocin peptide is administered simultaneously with IL-6, but in separate unit doses or formulations. In some embodiments, the oxytocin peptide and IL-6 are administered sequentially. In some embodiments, IL-6 is administered to the subject in a first dose, and then the oxytocin peptide is administered to the subject in a second dose. In some of these embodiments, the oxytocin peptide is administered about 1 minute to about 4 hours after the administration of IL-6. In some of these embodiments, the oxytocin peptide is administered approximately 1 minute to 4 hours, 10 minutes to 4 hours, 10 minutes to 3 hours, 10 minutes to 2 hours, 10 minutes to 1 hour, 10 minutes to 30 minutes, 20 minutes to 4 hours, 20 minutes to 3 hours, 20 minutes to 2 hours, 20 minutes to 1 hour, 30 minutes to 4 hours, 30 minutes to 3 hours, 30 minutes to 2 hours, or 30 minutes to 1 hour after the administration of IL-6. In some of these embodiments, the oxytocin peptide is administered approximately 1 minute, 10 minutes, 15 minutes, 20 minutes, 30 minutes, 45 minutes, 60 minutes, 90 minutes, 120 minutes, 150 minutes, 180 minutes, 210 minutes, or 240 minutes after the administration of IL-6. In some of these embodiments, the oxytocin peptide is administered approximately 10 minutes, 15 minutes, 20 minutes, or 30 minutes after the administration of IL-6. In one embodiment, the oxytocin peptide is administered to the subject first, followed by the administration of IL-6. In some embodiments, the subject is human.In some of these embodiments, magnesium ions are administered simultaneously with oxytocin peptide and / or IL-6, either before or after either or both of the oxytocin peptide and IL-6.

[0097] Oxytocin peptide and magnesium ions may be administered to subjects in need via the same route or via different routes. In some embodiments, oxytocin peptide is administered via craniofacial mucosal administration (e.g., intranasal, buccal, sublingual, or intraocular administration). In one embodiment, both oxytocin peptide and magnesium ions are administered intranasally as the same formulation. In one embodiment, oxytocin peptide is administered via the craniofacial mucosa, and magnesium ions are administered systemically, for example, intravenously, intramuscularly, orally, subcutaneously, or intrathecally.

[0098] In some embodiments, the oxytocin peptide is administered via intranasal administration. In some embodiments, the oxytocin peptide and magnesium ions are administered via intranasal administration. The oxytocin peptide and / or magnesium ions may be administered to the mucosal tissue within the nasal cavity using a suitable device for intranasal delivery (e.g., a transnasal delivery device described herein). Suitable regions within the nasal cavity include, but are not limited to, the lower two-thirds or upper one-third of the nasal cavity or the entire nasal cavity. In some embodiments, the oxytocin peptide and / or magnesium ions are administered to the upper one-third of the nasal cavity. In some embodiments, the oxytocin peptide and / or magnesium ions are administered to the lower two-thirds of the nasal cavity. In some embodiments, the oxytocin peptide and / or magnesium ions are explicitly administered to reach both the lower two-thirds and the upper one-third of the nasal cavity. In some embodiments, a method is provided for treating autism spectrum disorder, one or more symptoms associated with autism spectrum disorder, or a disorder exhibiting one or more symptoms associated with autism spectrum disorder, the method comprising the step of administering an effective dose of oxytocin peptide and magnesium ions intranasally to a subject in need, wherein the simultaneous administration of the oxytocin peptide and magnesium ions results in a synergistic or enhanced effect. In some embodiments, the method is for treating social and communication deficits or anxiety.

[0099] In some embodiments, IL-6 is administered to a subject by any of the methods described herein, via intranasal administration. IL-6 may be administered into the mucosal tissue of the nasal cavity using a device suitable for intranasal delivery (e.g., a nasal delivery device described herein). In some embodiments, IL-6 is administered systemically, for example, intravenously, intramuscularly, orally, subcutaneously, or intrathecally.

[0100] In some embodiments, the oxytocin peptide is human oxytocin consisting of Cys-Tyr-Ile-Gln-Asn-Cys-Pro-Leu-Gly (SEQ ID NO: 1). In some embodiments, the effective dose of the oxytocin peptide is about 0.5 μg to about 2000 μg. In some embodiments, the effective dose of the oxytocin peptide is about 0.5 μg to about 1000 μg, about 1 μg to about 1000 μg, or about 1 μg to about 2000 μg. In some embodiments, the effective dose of oxytocin peptide is approximately 4 μg to 1000 μg, 8 μg to 1000 μg, 8 μg to 800 μg, 8 μg to 500 μg, 8 μg to 400 μg, 8 μg to 300 μg, 8 μg to 200 μg, 8 μg to 100 μg, 8 μg to 80 μg, 8 μg to 50 μg, 10 μg to 1000 μg, 10 μg to 500 μg, 10 μg to 200 μg, 10 μg to 100 μg, 16 μg to 1000 μg, 16 μg to 800 μg, 16 μg to 500 μg, and 16 μg to 400 μg. g, about 16μg to about 200μg, about 16μg to about 160μg, about 16μg to about 120μg, about 16μg to about 80μg, about 20μg to about 100 0μg, about 20μg to about 800μg, about 20μg to about 500μg, about 20μg to about 200μg, about 20μg to about 100μg, about 30μg to about 1 The values ​​are approximately 000 μg, 30 μg to 500 μg, 30 μg to 300 μg, 30 μg to 120 μg, 30 μg to 90 μg, 50 μg to 1000 μg, 50 μg to 500 μg, 50 μg to 250 μg, 50 μg to 100 μg, or 50 μg to 80 μg. In some embodiments, the effective dose of oxytocin peptide is about 8 μg, about 16 μg, about 32 μg, about 48 μg, about 64 μg, about 80 μg, about 96 μg, about 128 μg, about 256 μg, about 10 μg, about 20 μg, about 30 μg, about 40 μg, about 50 μg, about 60 μg, about 70 μg, about 80 μg, about 90 μg, about 100 μg, about 120 μg, about 150 μg, about 200 μg, about 400 μg, about 600 μg, about 800 μg, or about 100 μg. In preferred embodiments, the effective dose of oxytocin peptide is about 8 μg to about 120 μg, about 15 μg to about 120 μg, about 30 μg to about 120 μg, or about 66 μg.

[0101] In some embodiments, the effective dose of oxytocin peptide is about 0.25 IU to about 1000 IU. In some embodiments, the effective dose of oxytocin peptide is about 0.25 IU to about 500 IU, about 0.5 IU to about 500 IU, or about 0.5 IU to about 1000 IU. In some embodiments, the effective dose of oxytocin peptide is approximately 2 IU to 500 IU, approximately 4 IU to 500 IU, approximately 4 IU to 400 IU, approximately 4 IU to 250 IU, approximately 4 IU to 200 IU, approximately 4 IU to 150 IU, approximately 4 IU to 100 IU, approximately 4 IU to 50 IU, approximately 4 IU to 40 IU, approximately 4 IU to 25 IU, approximately 5 IU to 500 IU, approximately 5 IU to 250 IU, approximately 5 IU to 100 IU, approximately 5 IU to 50 IU, approximately 8 IU to 500 IU, approximately 8 IU to 400 IU, approximately 8 IU to 250 IU, and approximately 8 IU to 200 IU. These ranges are approximately 8IU to 100IU, 8IU to 80IU, 8IU to 60IU, 8IU to 40IU, 10IU to 500IU, 10IU to 400IU, 10IU to 250IU, 10IU to 100IU, 10IU to 50IU, 15IU to 500IU, 15IU to 250IU, 15IU to 150IU, 15IU to 60IU, 15IU to 45IU, 25IU to 500IU, 25IU to 250IU, 25IU to 125IU, 25IU to 50IU, or 25IU to 40IU. In some embodiments, the effective dose of oxytocin peptide is about 4 IU, about 8 IU, about 16 IU, about 24 IU, about 32 IU, about 40 IU, about 48 IU, about 64 IU, about 128 IU, about 5 IU, about 10 IU, about 15 IU, about 20 IU, about 25 IU, about 30 IU, about 35 IU, about 40 IU, about 45 IU, about 50 IU, about 60 IU, about 75 IU, about 100 IU, about 200 IU, about 300 IU, about 400 IU, or about 50 IU. In preferred embodiments, the effective dose of oxytocin peptide is about 4 IU to about 60 IU, about 7.5 IU to about 60 IU, about 15 IU to about 60 IU, or about 30 IU.

[0102] The dose or amount of oxytocin in combination therapy is effective in one embodiment to provide clinically measurable improvement in the symptoms of autism spectrum disorder or related disorders. The combination of oxytocin and magnesium ions provides a synergistic or enhanced effect in improving autism spectrum disorder or related disorders. In some embodiments, oxytocin is administered at a dose below the therapeutically effective dose compared to the dose of oxytocin administered as a monotherapy. The dose of oxytocin as a monotherapy is partially dependent on the route of administration. Therefore, the dose of oxytocin in combination therapies described herein is also partially dependent on the route of administration.

[0103] The optimal dose of magnesium ions may depend on other factors such as the specific disorder or symptoms, the type of desired synergistic or enhanced effect, and the route of administration. The optimal dose can be measured in terms of the total amount of magnesium ions administered or the concentration of magnesium ions in the administered formulation. In some embodiments, the effective dose of magnesium ions administered is about 50 μg to about 68 mg. In some embodiments, the effective dose of magnesium ions administered is about 50 μg to about 34 mg, or about 1 mg to about 3 mg. In some embodiments, the effective dose of magnesium ions administered is about 1.3 mg, or about 2.6 mg. In some embodiments, the effective dose of magnesium ions administered is about 1.2 mg, or about 2.4 mg. In some embodiments, the effective dose of magnesium ions administered is approximately 50 μg to 17 mg, 50 μg to 8 mg, 50 μg to 4 mg, 50 μg to 2 mg, 50 μg to 1 mg, 50 μg to 500 μg, 100 μg to 68 mg, 100 μg to 34 mg, 100 μg to 17 mg, and 100 μg to 8 mg. , about 100 μg to about 4 mg, about 100 μg to about 2 mg, about 100 μg to about 1 mg, about 100 μg to about 500 μg, about 200 μg to about 68 mg, about 200 μg to about 34 mg, Approximately 200μg to approximately 17mg, approximately 200μg to approximately 8mg, approximately 200μg to approximately 4mg, approximately 200μg to approximately 2mg, approximately 200μg to approximately 1mg, approximately 200μg to approximately 500μg, approximately 5 00μg~about 68mg, about 500μg~about 34mg, about 500μg~about 17mg, about 500μg~about 8mg, about 500μg~about 5mg, about 500μg~about 4mg, about 500 μg~about 3mg, about 500μg~about 2mg, about 500μg~about 1mg, about 1mg~about 68mg, about 1mg~about 34mg, about 1mg~about 17mg, about 1mg~about 8mg, about 1 The dosage ranges are approximately 1 mg to 6 mg, 1 mg to 5 mg, 1 mg to 4 mg, 1 mg to 3 mg, 1 mg to 2 mg, 1.5 mg to 8 mg, 1.5 mg to 6 mg, 1.5 mg to 5 mg, 1.5 mg to 4 mg, 1.5 mg to 3 mg, 1.5 mg to 2 mg, 1.3 mg to 2.6 mg, or 1.2 mg to 2.4 mg.In some embodiments, magnesium ions are provided using magnesium salts (e.g., magnesium citrate and / or magnesium chloride).

[0104] In some embodiments, the administered magnesium salt comprises magnesium chloride, and the effective dose of magnesium salt is approximately 0.48 mg to approximately 600 mg of magnesium chloride hexahydrate (MgCl2·6H2O,MW2O3.3). In some embodiments, the effective doses of magnesium chloride hexahydrate are approximately 0.48 mg to approximately 300 mg, approximately 0.5 mg to approximately 150 mg, approximately 0.5 mg to approximately 75 mg, approximately 5 mg to approximately 150 mg, approximately 5 mg to approximately 75 mg, approximately 5 mg to approximately 50 mg, approximately 10 mg to approximately 600 mg, approximately 10 mg to approximately 300 mg, approximately 10 mg to approximately 150 mg, approximately 10 mg to approximately 75 mg, approximately 10 mg to approximately 50 mg, approximately 10 mg to approximately 30 mg, or approximately 12 mg to approximately 24 mg. In some preferred embodiments, the effective dose of magnesium chloride hexahydrate is about 6 mg, about 12 mg, about 18 mg, about 24 mg, or about 30 mg.

[0105] In some embodiments, the magnesium salt administered is magnesium citrate, and the effective dose of the magnesium salt is approximately 0.48 mg to approximately 600 mg of magnesium citrate. In some embodiments, the effective dose of magnesium citrate (e.g., anhydrous dibasic magnesium citrate, molecular weight: 214.4) is approximately 0.48 mg to approximately 300 mg, approximately 0.5 mg to approximately 150 mg, approximately 0.5 mg to approximately 75 mg, approximately 5 mg to approximately 150 mg, approximately 5 mg to approximately 75 mg, approximately 5 mg to approximately 50 mg, approximately 10 mg to approximately 600 mg, approximately 10 mg to approximately 300 mg, approximately 10 mg to approximately 150 mg, approximately 10 mg to approximately 75 mg, approximately 10 mg to approximately 50 mg, approximately 10 mg to approximately 30 mg, or approximately 12 mg to approximately 24 mg. In some preferred embodiments, the effective dose of magnesium citrate (e.g., anhydrous dibasic magnesium citrate, molecular weight: 214.4) is about 6 mg, about 12 mg, about 18 mg, about 24 mg, or about 30 mg. In some embodiments, the effective dose of magnesium citrate is approximately 0.48 mg to 12 mg, approximately 0.5 mg to 10 mg, approximately 0.5 mg to 8 mg, approximately 0.5 mg to 5 mg, approximately 0.5 mg to 2.5 mg, approximately 0.5 mg to 1 mg, approximately 1 mg to 10 mg, approximately 1 mg to 8 mg, approximately 1 mg to 5 mg, approximately 1 mg to 2 mg, approximately 2 mg to 10 mg, approximately 2 mg to 8 mg, approximately 2 mg to 6 mg, approximately 2 mg to 4 mg, approximately 3 mg to 10 mg, approximately 4 mg to 10 mg, approximately 4 mg to 8 mg, approximately 4 mg to 6 mg, approximately 5 mg to 10 mg, approximately 5 mg to 8 mg, approximately 5 mg to 7 mg, approximately 5 mg to 6 mg, approximately 6 mg to 10 mg, approximately 6 mg to 8 mg, or approximately 6 mg to 7 mg. When other magnesium salts are used in place of magnesium citrate, the effective dose of that magnesium salt provides an amount of magnesium ions equal to that provided by magnesium citrate.

[0106] It is intended and understood that each and all doses of magnesium ions described herein may be combined with each and all doses of oxytocin peptides described herein, as if each and all combinations were described individually. For example, in some embodiments, the effective dose of oxytocin peptide is about 0.5 μg to about 2000 μg, and the effective dose of magnesium ions is about 50 μg to about 68 mg of magnesium. In some embodiments, the effective dose of oxytocin peptide is about 15 μg to about 120 μg (e.g., about 60 μg or about 66 μg), and the effective dose of magnesium ions is equal to the amount of magnesium ions provided by about 10 mg to about 30 mg (e.g., about 12 mg or about 24 mg) of magnesium citrate.

[0107] In some embodiments, methods are provided for treating autism spectrum disorder, one or more symptomatic disorders associated with autism spectrum disorder, social and communication deficits, or anxiety, the methods comprising the step of administering an effective dose of oxytocin peptide and magnesium ions (e.g., by intranasal administration) to a subject in need thereof, wherein the weight ratio of the dose of oxytocin peptide administered to the dose of magnesium ions administered is about 1:1 to about 1:1000, preferably about 1:2 to about 1:200, more preferably about 1:20, about 1:30, about 1:35, about 1:40, about 1:45, about 1:50, about 1:60, or any of the OT / Mg(w) ratios described herein with respect to magnesium-containing oxytocin peptide formulations or compositions. In some embodiments, methods are provided for treating autism spectrum disorder, one or more symptomatic disorders associated with autism spectrum disorder, social and communication deficits, or anxiety, the method comprising the step of administering an effective dose of oxytocin peptide and magnesium ions (e.g., by intranasal administration) to a subject in need thereof, wherein the molar ratio of the dose of oxytocin peptide administered to the dose of magnesium ions administered is The OT / Mg(m) ratio is approximately 1:40 to approximately 1:40000, preferably approximately 1:80 to approximately 1:8000, more preferably approximately 1:175, approximately 1:280, approximately 1:500, approximately 1:560, approximately 1:800, approximately 1:1000, approximately 1:1100, approximately 1:1200, approximately 1:1400, approximately 1:1600, approximately 1:1700, approximately 1:1800, approximately 1:2000, approximately 1:2400, approximately 1:3000, or any of the OT / Mg(m) ratios described herein with respect to magnesium-containing oxytocin peptide formulations or compositions. In some of these embodiments, the oxytocin peptide is human oxytocin consisting of Cys-Tyr-Ile-Gln-Asn-Cys-Pro-Leu-Gly (SEQ ID NO: 1). In some of these embodiments, the magnesium ions are provided by magnesium citrate and / or magnesium chloride.In some of these embodiments, the deficiencies in sociality and communication include impaired communication skills and / or dysfunction in social interaction, lack of eye contact, and / or inability to form and / or maintain social relationships.

[0108] In one embodiment, a method is provided for treating autism spectrum disorder, a disorder exhibiting one or more symptoms associated with autism spectrum disorder, social and communication deficits, or anxiety, the method comprising the step of intranasally administering to a subject in need an oxytocin peptide in a dose of about 0.5 μg to about 2000 μg (e.g., about 8 μg to about 300 μg, about 15 μg to about 120 μg, or about 66 μg) and magnesium or magnesium ions in a dose of about 50 μg to about 68 mg, about 50 μg to about 34 mg, about 1 mg to about 3 mg, about 1.3 mg, or about 2.6 mg. In one embodiment, the method comprises the step of intranasally administering to a subject in need an effective amount of a magnesium-containing oxytocin peptide preparation or composition described herein. In one embodiment, the method comprises the step of administering an effective amount of oxytocin peptide and magnesium ions intranasally to a subject in need in a liquid formulation with a volume of about 5 μL to about 1000 μL. In one embodiment, the method comprises the step of administering an effective dose of a magnesium-containing oxytocin peptide formulation or composition containing about 0.01 mg / mL to about 16 mg / mL (e.g., about 0.1 mg / mL and about 16 mg / mL) of oxytocin and about 1 mg / mL to about 30 mg / mL of magnesium or magnesium ions intranasally to the subject in need. In one embodiment, the method includes administering an effective dose of a magnesium-containing oxytocin peptide preparation containing about 0.01 mg / mL to about 16 mg / mL (e.g., about 0.1 mg / mL to about 16 mg / mL or about 0.15 mg / mL to about 1.5 mg / mL) of oxytocin and about 1 (by weight) to about 25 (by weight) (e.g., about 1% to about 15% or about 10% to about 14%) of magnesium citrate into the nasal cavity of a subject in need.In one embodiment, the method includes administering an effective dose of a magnesium-containing oxytocin peptide preparation containing about 5 IU / mL to about 8000 IU / mL (e.g., about 50 IU / mL to about 8000 IU / mL or about 75 IU / mL to about 750 IU / mL) of oxytocin and about 1 (by weight) to about 25 (by weight) (e.g., about 1% to about 15%, about 10% to about 14%, or about 12%) of magnesium citrate into the nasal cavity of a subject in need. In one embodiment, the method includes administering an effective dose of a magnesium-containing oxytocin peptide preparation containing about 0.01 mg / mL to about 16 mg / mL (e.g., about 0.1 mg / mL and about 16 mg / mL or about 0.15 mg / mL and about 1.5 mg / mL) of oxytocin and about 1 (by weight) to about 25 (by weight) (e.g., about 1% to about 15%, about 8% to about 12%, or about 10%) of magnesium chloride hexahydrate into the nasal cavity of a subject in need. In one embodiment, the method comprises administering an effective dose of a magnesium-containing oxytocin peptide preparation containing about 5 IU / mL to about 8000 IU / mL (e.g., about 50 IU / mL to about 8000 IU / mL or about 75 IU / mL to about 750 IU / mL) of oxytocin and about 1 (by weight) to about 25 (by weight) (e.g., about 1% to about 15%, about 8% to about 12%, or about 10%) of magnesium chloride hexahydrate into the nasal cavity of a subject in need.

[0109] In some embodiments, an effective dose of oxytocin peptide and magnesium ions includes about 0.5 μg (or 0.25 IU) to about 2000 μg (or 1000 IU) of oxytocin peptide administered in an aqueous solution containing about 0.1% to about 2.8% (w / v) magnesium. In some embodiments, an effective dose of oxytocin peptide and magnesium ions includes about 8 μg (or 4 IU) to about 1000 μg (or 500 IU) of oxytocin peptide administered in an aqueous solution containing about 0.11% to about 1.65% (w / v) magnesium. In some embodiments, an effective dose of oxytocin peptide and magnesium ions comprises about 15 μg (or 7.5 IU) to about 120 μg (or about 60 IU) (e.g., about 60 μg or 30 IU) of oxytocin peptide administered in an aqueous solution containing about 1.1% to about 1.6% (e.g., about 1.2% or about 1.35%) magnesium. In one embodiment, an effective dose of oxytocin peptide and magnesium ions comprises about 60 μg (or 30 IU) of oxytocin peptide administered in an aqueous solution containing about 1.2% or about 1.35% magnesium.

[0110] In some embodiments, an effective dose of oxytocin peptide and magnesium ions is administered intranasally in a liquid formulation, with the volume of the administered liquid formulation being approximately 5 μL to approximately 1000 μL. In some embodiments, the administered volume is approximately 5 μL to approximately 500 μL, approximately 5 μL to approximately 250 μL, approximately 5 μL to approximately 100 μL, approximately 5 μL to approximately 50 μL, approximately 10 μL to approximately 1000 μL, approximately 10 μL to approximately 500 μL, approximately 10 μL to approximately 250 μL, approximately 10 μL to approximately 100 μL, approximately 25 μL to approximately 1000 μL, approximately 25 μL to approximately 500 μL, approximately 25 μL to approximately 250 μL, approximately 25 μL to approximately 100 μL, and approximately 50 μL to approximately 1 The available sizes are 000 μL, approximately 50 μL to 750 μL, approximately 50 μL to 500 μL, approximately 50 μL to 450 μL, approximately 50 μL to 400 μL, approximately 50 μL to 350 μL, approximately 50 μL to 300 μL, approximately 50 μL to 250 μL, approximately 50 μL to 200 μL, approximately 50 μL to 150 μL, approximately 100 μL to 500 μL, approximately 100 μL to 400 μL, approximately 100 μL to 300 μL, or approximately 100 μL to 200 μL. In some embodiments, the administered volume is about 50 μL, about 100 μL, about 150 μL, about 200 μL, about 250 μL, about 300 μL, about 350 μL, about 400 μL, about 450 μL, or about 500 μL. In some embodiments, an effective dose of oxytocin peptide and magnesium ions is administered intranasally in a liquid formulation contained in a nasal device described herein.

[0111] The combinations of oxytocin peptide and magnesium ions described herein may be used to treat any oxytocin-treated social and communication deficiencies (e.g., impairments in communication skills and / or social interaction, lack of eye contact, and / or inability to form and / or maintain social relationships). Accordingly, a method for treating social and communication deficiencies is provided, comprising the step of administering an effective dose of oxytocin peptide and magnesium ions to a subject in need, where the social and communication deficiency is defined as impairments in communication skills and / or social interaction, lack of eye contact, and / or inability to form and / or maintain social relationships. In one embodiment, the method comprises the step of administering an effective dose of oxytocin peptide and magnesium ions intranasally to a subject in need. In some embodiments, the molar ratio of the amount of oxytocin peptide to the amount of magnesium or magnesium ions is about 1:175, about 1:280, about 1:560, about 1:1100, about 1:1700, or about 1:2000 (including any range between these ratios).

[0112] In one embodiment, a method is provided for treating autism spectrum disorder, a disorder exhibiting one or more symptoms associated with autism spectrum disorder, social and communication deficits, or anxiety, the method comprising the step of administering an effective dose of oxytocin peptide and magnesium ions intranasally to a subject in need (e.g., a human or veterinary patient), wherein the simultaneous administration of the oxytocin peptide and magnesium ions results in a synergistic or enhanced effect. In some embodiments, the oxytocin peptide is human oxytocin consisting of Cys-Tyr-Ile-Gln-Asn-Cys-Pro-Leu-Gly (SEQ ID NO: 1). In some embodiments, the effective dose of oxytocin peptide is about 0.5 μg (or 0.25 IU) to about 2000 μg (or 1000 IU), preferably about 8 μg (or 4 IU) to about 1000 μg (or 500 IU), and more preferably about 15 μg (or 7.5 IU) to about 120 μg (or 60 IU). In some embodiments, the effective dose of magnesium ions is about 50 μg to about 68 mg. In some embodiments, magnesium ions are provided using magnesium salts (e.g., magnesium chloride and / or magnesium citrate) administered in amounts that provide about 50 μg to about 68 mg of magnesium. In some embodiments, the effective dose of magnesium ions is provided by using about 0.48 mg to about 600 mg of magnesium citrate. In some embodiments, the effective dose of magnesium ions is provided by using about 0.42 mg to about 540 mg of magnesium chloride hexahydrate. In some embodiments, an effective dose of oxytocin peptide and magnesium ions comprises about 15 μg (or 7.5 IU) to about 120 μg (or 60 IU) (e.g., about 60 μg or 30 IU) of oxytocin peptide administered in an aqueous solution containing about 1.1% to about 1.54% (e.g., about 1.2% or about 1.35%) (w / v) magnesium.In some embodiments, an effective dose of oxytocin peptide and magnesium ions comprises about 10 μg to about 120 μg (e.g., about 66 μg) of oxytocin peptide administered in an aqueous solution containing about 10% to about 14% (e.g., about 12%) (w / v) magnesium citrate.

[0113] kit A kit for performing any of the methods described herein is provided herein. The kit is provided for use in the treatment of autism spectrum disorder, one or more symptomatic disorders associated with autism spectrum disorder, social and communication deficits, or anxiety. In some embodiments, the kit comprises an oxytocin peptide and magnesium ions in suitable packaging, wherein the oxytocin peptide and magnesium ions are in amounts that produce a synergistic or enhanced effect when used in the treatment of autism spectrum disorder, one or more symptomatic disorders associated with autism spectrum disorder, social and communication deficits, or anxiety, as well as a device for craniofacial mucosal administration (e.g., intranasal administration). The kit may further comprise a protease inhibitor and / or at least one absorption enhancer. The kit may further comprise IL-6. Other kits may further comprise instructions that provide information to the user and / or healthcare provider for performing any of the methods described herein. The kit may further include reagents / tools for measuring IL-6 levels in subjects, as well as instructions for predicting the effectiveness of nasal oxytocin and magnesium ions, if necessary.

[0114] Also provided are kits comprising a magnesium-containing oxytocin peptide preparation and suitable packaging materials, incorporated into a device for craniofacial mucosal administration (e.g., an intranasal administration device such as a nasal pump device). The kit may further include instructions for administering the magnesium-containing oxytocin peptide preparation to a subject requiring the preparation.

[0115] Instructions for using a kit to carry out the present invention typically describe how to use the contents of the kit to carry out the method of the present invention. Instructions supplied with a kit of the present invention are typically written instructions on a label or accompanying documents (e.g., a paper sheet included in the kit), but machine-readable instructions (e.g., instructions on a magnetic disk or optical storage disk) are also acceptable. [Examples]

[0116] The present invention may be further understood by referring to the following examples, which are provided for illustrative purposes and are not intended to limit the scope of the invention.

[0117] Example 1: Exemplary preparation of a magnesium-containing oxytocin peptide formulation Example 1A The hypertonic drug product formulation, targeting pH 4.5, consists of oxytocin USP (150 IU / mL); magnesium chloride USP (as hexahydrate or anhydrous salt); citrate USP (as anhydrous or monohydrate); sodium hydroxide NF; and sterile water for injection USP. The quantitative composition is provided in Table 1. The molar ratio of oxytocin to magnesium ions in the formulation is approximately 1:1679. All components meet the requirements of the corresponding monograph pharmacopoeias (USP / NF). [Table 1]

[0118] The drug products are manufactured by dissolving the ingredients in sterile water for injection, sterilizing and filtering the solution, and filling them into snap-on vials using a preservative-free pump, and are tested in general accordance with the FDA's July 2002 guidelines for nasal sprays.

[0119] In one example, a 10 L batch of magnesium-containing oxytocin preparation according to the composition provided in Table 1 was prepared as follows: The preparation container was filled with water to approximately 60% of the required batch volume. The required amounts of sodium chloride, citric acid, and magnesium chloride hexahydrate were added in the following order while stirring at ambient temperature. These materials dissolved readily. Heating was not necessary; only gentle stirring was required. The pH of the solution was adjusted to 4.5 by adding 1N NaOH (in case of excess titration, back titration to pH 4.5 with 10% HCl was possible). The required amount of oxytocin was added and stirred until dissolved. Water was added until the final weight / volume of the batch was reached. The solution was stirred until homogeneous.

[0120] Example 1B The isotonic drug formulation, targeting pH 4.5, consists of oxytocin USP (150 IU / mL), magnesium citrate, sodium chloride USP, sodium acetate trihydrate USP, glacial acetic acid USP, and sterile water for injection USP. The quantitative composition is provided in Table 2. The molar ratio of oxytocin to magnesium ions in the formulation is approximately 1:1992. The target pH of 4.5 is selected based on optimal formulation stability at or near pH 4.5 (Hawe et al., Pharmaceut. Res. 2009, 26:1679-1688). All components meet the requirements of the corresponding monograph pharmacopoeias (USP / NF).

[0121] To prepare the oxytocin stock solution, add lyophilized oxytocin (2 mg) to 1 mL of water (USP), 0.9% physiological saline, or phosphate-buffered saline in a 5 mL glass container. Stir the solution until all the oxytocin is dissolved, and adjust the pH to 3.5–8.5 to produce 1 mL of 2 mg / mL (approximately 1000 IU / mL) liquid oxytocin preparation.

[0122] When used as a clinical material, oxytocin and excipients are manufactured according to current Good Manufacturing Practices, sterilized (sterile filtration using a 0.2 micron membrane filter), filled into glass storage bottles, and sealed with a pump actuator. By increasing or decreasing the amount of oxytocin, various formulation concentrations can be produced from this example. Approximately 10 doses of oxytocin can be obtained from this 1 mL batch volume. [Table 2]

[0123] Example 2: A rat model of social behavior Rats were intranasally treated with either physiological saline, a combination of 10 μg oxytocin, 12% magnesium citrate, and 10 μg oxytocin (oxytocin to magnesium ion molar ratio of approximately 1:1127), or a 20 μl (10 μl / nasal cavity) solution containing 12% magnesium citrate. Eight rats were used in each treatment group. Forty minutes after intranasal drug administration, two animals from the same treatment group were paired and placed in a test chamber, and their social behaviors (sniffing, following, crawling, allogrooming [grooming a partner], and play fighting) were recorded for 10 minutes. The time spent on social interaction is shown in Figure 1. The results provide evidence of an enhanced effect of the combination of 12% magnesium citrate and 10 μg oxytocin on improving social behavior.

[0124] Example 3: Anxiety-inducing rat model Example 3A Rats were intranasally treated with either physiological saline, a combination of 10 μg oxytocin, 12% magnesium citrate, and 10 μg oxytocin (oxytocin to magnesium ion molar ratio of approximately 1:1127), or a 20 μl (10 μl / nasal cavity) solution containing 12% magnesium citrate. Eight rats were used in each treatment group. Fifty minutes after intranasal drug administration, the animals were placed in a radial arm maze, and their anxiety was assessed by the number of times the animals entered an open arm within a 5-minute period. The observed number of times the animals entered an open arm is shown in Figure 2. The results provide evidence of a synergistic effect of the combination of 12% magnesium citrate and 10 μg oxytocin on reducing anxiety.

[0125] Example 3B Rats were intranasally treated with 20 μl (10 μl / nasal cavity) of a solution containing physiological saline, 3% magnesium citrate, 6% magnesium citrate, 16 μg oxytocin, 10 μg oxytocin, a combination of 3% magnesium citrate and 16 μg oxytocin (oxytocin to magnesium ion molar ratio of approximately 1:176), or a combination of 6% magnesium citrate and 10 μg oxytocin (oxytocin to magnesium ion molar ratio of approximately 1:563). Eight rats were used in each treatment group. Thirty minutes after intranasal drug administration, the animals were exposed to 5 minutes of elevated platform stress, and immediately afterward, placed in an elevated platform plus maze for 5 minutes. Their anxiety was assessed by the number of times the animals entered the open arms during the 5 minutes. The observed number of times the animals entered the open arms is shown in Figures 3A and 3B. Animals treated with a combination of 3% magnesium citrate and 16 μg oxytocin (oxytocin to magnesium ion molar ratio of approximately 1:176) exhibited lower anxiety than animals treated with either 3% magnesium citrate alone or 16 μg oxytocin alone, as evidenced by an increased number of times they entered open arms. In contrast, 6% magnesium citrate and 10 μg oxytocin Animals treated with an oxytocin combination (oxytocin to magnesium ions in a molar ratio of approximately 1:563) showed a 6% reduction in the number of times they entered the open arm, as indicated by this reduction. Animals treated with either magnesium citrate alone or 10 μg oxytocin alone exhibited greater anxiety.

[0126] To further assess anxiety, the latency to enter the open arm, the time spent in the open arm, and the number of times the closed arm was entered were determined.

[0127] Repeat the experiment with further amounts of magnesium citrate and oxytocin (e.g., including 6% magnesium citrate alone, 20 μg oxytocin alone, and a combination of 6% magnesium citrate and 20 μg oxytocin (with a molar ratio of oxytocin to magnesium ions of approximately 1:281)).

[0128] Example 4: Single Subject Case Study Subjects diagnosed with autism spectrum disorder (e.g., children) are administered a liquid formulation containing 12-24 IU of oxytocin intranasally every morning and evening for three days. The subjects' social functioning and anxiety are assessed. After a four-day rest period, the subjects are administered a liquid formulation containing 3%-12% magnesium citrate intranasally every morning and evening for three days, and their social functioning and anxiety are assessed. After a four-day rest period, the subjects are administered a liquid formulation containing a combination of 12-24 IU of oxytocin and 3%-12% magnesium citrate intranasally every morning and evening for three days, and their social functioning and anxiety are assessed.

[0129] Example 5: Human Clinical Trial The efficacy of a 6-week course of intranasal treatment twice daily with a combination of oxytocin and magnesium will be tested in male and female subjects aged 18–55 years diagnosed with autism spectrum disorder, using a placebo-controlled, randomized, double-blind, parallel design. The primary efficacy endpoint is the change in social interaction scores, as measured by the Autism Diagnostic Observation Schedule-II, before and after the double-blind treatment period. Secondary endpoints will consist of one or more of the following: (1) Changes in communication and restricted repetitive behavior scores as measured by the Autism Diagnostic Observational Scale II, before and at the completion of the double-blind treatment period; (2) Changes in anxiety as measured by the State Trait Anxiety Scale, assessed before and at the completion of the double-blind treatment period; (3) Changes in depression as measured by the Center for Epidemiologic Studies Depression Scale, assessed before and at the completion of the double-blind treatment period; (4) Changes in gaze towards the atmosphere of the environment, as evaluated before and at the completion of the double-blind treatment period; (5) Changes in facial and vocal expression as analyzed from videos recorded every two weeks during the clinical trial period; and (6) Changes in overall clinical impression and overall functional assessment scales, which are evaluated every two weeks during the trial period.

[0130] The research inclusion criteria consist of the following: 1) Diagnosed with autism spectrum disorder based on DSM-V; 2) Exceeding the cutoff for qualitative abnormalities in reciprocal social interaction (domain A) in the Autism Diagnostic Observational-Revised Edition; and 3) Verbal IQ above 85 and Full Scale IQ above 80 as measured by the Wechsler Adult Intelligence Scale III.

[0131] The research exclusion criteria consist of the following: 1) Major psychiatric diagnoses other than those defined in inclusion criterion 1); 2) Current instability resulting from co-occurring psychiatric diagnoses; 3) History of changes in psychotropic drug therapy or dosage within one month of randomization; 4) Currently being treated with more than two categories of psychotropic drugs; 5) Currently receiving treatment with atomoxetine or methylphenidate; 6) History of continuous oxytocin treatment; 7) History of sensitivity to oxytocin; 8) A history of seizures or traumatic brain injury involving loss of consciousness for more than 5 minutes; and 9) A history of alcohol dependence, substance abuse, or substance addiction.

[0132] Example 6: Effects of oxytocin and magnesium on social anxiety disorder Subjects meeting the criteria of the Diagnostic and Statistical Manual of Mental Disorders, 4th Edition, relating to Generalized Social Phobia, will be randomized to receive one of the following treatments: intranasal placebo (saline) - Treatment A, oxytocin alone (30 IU) - Treatment B, magnesium alone (10%) - Treatment C, or oxytocin (30 IU) + magnesium (10%) - Treatment D.

[0133] After a one-week single-blind, placebo-equalization period, patients receive an 11-week double-blind course of treatment A, B, C, or D. Patients receive treatment twice daily, approximately 12 hours apart.

[0134] If necessary, serum IL-6 levels will be checked at the end of the one-week adjustment period and at the end of the 11-week cycle.

[0135] The number of respondents based on the Clinical Global Impression Global Improvement Item ("significantly improved" or "very improved"); the mean change from baseline in the Leibovitz Social Anxiety Scale total score is measured. If necessary, serum IL-6 levels are correlated with the degree of effectiveness to determine the effect of IL-6 as a predictive biomarker of effectiveness.

[0136] We will analyze the effects of the treatment group.

[0137] Exemplary Embodiments The present invention is further described by the following embodiments. Features of each of these embodiments can be combined with any of the other embodiments as needed and, if not permitted.

[0138] Embodiment 1. In one embodiment, a method is provided for treating autism spectrum disorder, one or more symptomatic disorders associated with autism spectrum disorder, social and communication deficits, or anxiety, the method comprising the step of administering an effective dose of oxytocin peptide and magnesium ions to a subject in need thereof, wherein the simultaneous administration of the oxytocin peptide and magnesium ions results in a synergistic or enhanced effect.

[0139] Embodiment 2. In a further embodiment of Embodiment 1, the oxytocin peptide is administered simultaneously with the magnesium ions.

[0140] Embodiment 3. In a further embodiment of Embodiment 1, the oxytocin peptide is administered before or after the administration of the magnesium ions.

[0141] Embodiment 4. In any further embodiment of Embodiments 1 to 3, the oxytocin peptide is administered via craniofacial mucosal administration.

[0142] Embodiment 5. In a further embodiment of Embodiment 4, the oxytocin peptide is administered via intranasal administration.

[0143] Embodiment 6. In a further embodiment of Embodiment 5, the oxytocin peptide and the magnesium ions are administered via intranasal administration.

[0144] Embodiment 7. In any further embodiment of Embodiments 1 to 6, the effective dose of the oxytocin peptide is about 0.5 μg to about 2000 μg.

[0145] Embodiment 8. In any one further embodiment of Embodiments 1 to 7, the effective dose of the magnesium ions is about 50 μg to about 68 mg.

[0146] Embodiment 9. In any further embodiment of Embodiments 1 to 8, the magnesium ions are provided using magnesium chloride and / or magnesium citrate.

[0147] Embodiment 10. In a further embodiment of Embodiment 1, the effective dose of the oxytocin peptide and the magnesium ions comprises about 15 μg to about 120 μg of the oxytocin peptide administered in an aqueous solution containing about 1.1% to about 1.6% (w / v) magnesium.

[0148] Embodiment 11. In a further embodiment of Embodiment 1, the effective dose of the oxytocin peptide and the magnesium ions has an oxytocin:magnesium molar ratio of about 1:40 to about 1:40000.

[0149] Embodiment 12. In any further embodiment of Embodiments 1 to 11, the method is for treating autism spectrum disorder.

[0150] Embodiment 13. In any further embodiment of Embodiments 1 to 11, the method is for treating one or more symptomatic disorders associated with autism spectrum disorder.

[0151] Embodiment 14. In a further embodiment of Embodiment 13, the disorder is social anxiety disorder, obsessive-compulsive disorder, social (pragmatic) communication disorder, neurodevelopmental disorder, attention deficit hyperactivity disorder, Prader-Willi syndrome, Timothy syndrome, fragile X syndrome, Rett syndrome, or Williams syndrome.

[0152] Embodiment 15. In any further embodiment of Embodiments 1 to 11, the method is for addressing deficiencies in social skills and communication.

[0153] Embodiment 16. In any further embodiment of Embodiments 1 to 11, the method is for the purpose of addressing anxiety.

[0154] Embodiment 17. In any further embodiment of Embodiments 1 to 16, the oxytocin peptide is human oxytocin (SEQ ID NO: 1).

[0155] Embodiment 18. In one embodiment, a method is provided for treating autism spectrum disorder, one or more symptomatic disorders associated with autism spectrum disorder, social and communication deficits, or anxiety, the method comprising the step of administering an effective dose of oxytocin peptide and magnesium ions to a subject in need, wherein the effective dose of oxytocin peptide and magnesium ions is administered intranasally in a liquid formulation, the volume of the liquid formulation administered is about 5 μL to about 1000 μL.

[0156] Embodiment 19. In a further embodiment of Embodiment 18, the effective dose of the oxytocin peptide is approximately 0.5 μg to approximately 2000 μg.

[0157] Embodiment 20. In a further embodiment of Embodiment 18, the effective dose of magnesium ions is approximately 50 μg to approximately 68 mg.

[0158] Embodiment 21. In a further embodiment of Embodiment 18, the effective dose of the oxytocin peptide and the magnesium ions comprises about 15 μg to about 120 μg of the oxytocin peptide administered in an aqueous solution containing about 1.1% to about 1.6% (w / v) magnesium.

[0159] Embodiment 22. In a further embodiment of Embodiment 18, the effective dose of the oxytocin peptide and the magnesium ions has an oxytocin:magnesium molar ratio of about 1:40 to about 1:40000.

[0160] Embodiment 23. In further embodiments of Embodiment 21 or 22, the volume of the liquid formulation to be administered is approximately 50 μL to approximately 200 μL.

[0161] Embodiment 24. In a further embodiment of Embodiment 23, the liquid formulation is administered using a metering nasal device in 1 to 4 units of approximately 50 μL / unit.

[0162] Embodiment 25. In any further embodiment of Embodiments 18 to 24, the method is for treating autism spectrum disorder.

[0163] Embodiment 26. In any further embodiment of Embodiments 18 to 24, the method is for treating one or more symptomatic disorders associated with autism spectrum disorder.

[0164] Embodiment 27. In a further embodiment of Embodiment 26, the disorder is social anxiety disorder, obsessive-compulsive disorder, social (pragmatic) communication disorder, neurodevelopmental disorder, attention deficit hyperactivity disorder, Prader-Willi syndrome, Timothy syndrome, fragile X syndrome, Rett syndrome, or Williams syndrome.

[0165] Embodiment 28. In any further embodiment of Embodiments 18 to 24, the method is for addressing deficiencies in social skills and communication.

[0166] Embodiment 29. In any one further embodiment of Embodiments 18 to 24, the method is for addressing anxiety.

[0167] Embodiment 30. In any further embodiment of Embodiments 18 to 29, the oxytocin peptide is human oxytocin (SEQ ID NO: 1).

[0168] Embodiment 31. In a further embodiment of Embodiment 18, the liquid formulation is contained within a device for intranasal administration.

[0169] Embodiment 32. In a further embodiment of Embodiment 31, the device for intranasal administration is a nasal pump device.

[0170] Embodiment 33. In a further embodiment of Embodiment 32, the nasal pump device comprises a storage bottle attached to a pump actuator.

[0171] Embodiment 34. In a further embodiment of Embodiment 33, the pump actuator measures to deliver a specific volume of approximately 50 μL.

[0172] Embodiment 35. In a further embodiment of Embodiment 32, the nasal pump device comprises a storage bottle attached to an aerozoizer.

[0173] Embodiment 36. In any further embodiment of Embodiments 32 to 35, the nasal pump device is as follows: (i) Filters to prevent backflow, (ii) Metal-free channels, and (iii) Plastic materials that are stable against gamma rays It must have one or more of the following.

[0174] Embodiment 37. In one embodiment, a composition is provided comprising an oxytocin peptide and a magnesium ion, wherein the oxytocin peptide and the magnesium ion are present in amounts that produce a synergistic or enhanced effect when used to treat anxiety.

[0175] Embodiment 38. In a further embodiment of Embodiment 37, the oxytocin peptide is human oxytocin (SEQ ID NO: 1).

[0176] Embodiment 39. In a further embodiment of Embodiment 37, the composition is a liquid formulation containing about 0.01 mg / mL to about 16 mg / mL of the oxytocin peptide.

[0177] Embodiment 40. In a further embodiment of Embodiment 37, the composition is a liquid formulation containing magnesium in an amount that provides about 3 mg / mL to about 30 mg / mL of magnesium.

[0178] Embodiment 41. In a further embodiment of Embodiment 37, the oxytocin peptide and the magnesium ions have a molar ratio of about 1:40 to about 1:40000.

[0179] Embodiment 42. In a further embodiment of Embodiment 41, the molar ratio is approximately 1:40 to approximately 1:800.

[0180] Embodiment 43. In a further embodiment of Embodiment 41, the molar ratio is approximately 1:800 to approximately 1:40000.

[0181] Embodiment 44. In any one further embodiment of Embodiments 37 to 43, a device for craniofacial mucosal administration is further included.

[0182] Embodiment 45. In a further embodiment of Embodiment 44, the oxytocin peptide and the magnesium ions are contained within the device for craniofacial mucosal administration.

[0183] Embodiment 46. In a further embodiment of Embodiment 45, the device is for intranasal administration.

[0184] While the aforementioned invention has been described in some detail for illustrative and illustrative purposes to clarify understanding, it will be apparent to those skilled in the art that certain modifications and alterations can be carried out without departing from the invention. Therefore, these descriptions and examples should not be construed as limiting the scope of the invention.

[0185] All patents, patent applications, documents, and articles cited herein are incorporated by direct reference.

Claims

[Claim 1] The invention as shown in the drawings.

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