Magnesium-containing oxytocin formulations and methods of use

Abstract

To provide magnesium-containing oxytocin formulations and methods of use.SOLUTION: Disclosed are methods and compositions for treatment of autism spectrum disorder, related disorders and symptoms of such disorders, comprising co-administration of an oxytocin peptide and magnesium ions. Co-administration of an oxytocin peptide and magnesium ions results in a synergistic or enhanced effect on reducing social and communication deficits in a patient suffering from an autism spectrum disorder. The invention may answer a need for an oxytocin peptide formulation capable of providing a more pronounced effect on the response of people having autism spectrum disorders and related disorders to oxytocin treatment that shows wide variability.SELECTED DRAWING: None

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Patent Application No. 62 / 321,654, filed April 12, 2016, the disclosure of which is incorporated herein by reference in its entirety.

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

[0003] BACKGROUND OF THEINVENTION Oxytocin is a naturally occurring nine amino acid neuropeptide produced primarily in the paraventricular and supraoptic nuclei of the mammalian hypothalamus. Oxytocin is released into the central nervous system via distributed neural pathways and into the peripheral circulation via the posterior pituitary gland. Intramuscular or intravenous injection of synthetic oxytocin (Pitocin®) is currently approved in the United States to produce or improve uterine contractions to promote vaginal delivery and to manage postpartum hemorrhage. Intranasal oxytocin (Syntocinon®) was approved in the United States from 1960 to 1997 to stimulate milk ejection and promote breastfeeding. Syntocinon® nasal spray was withdrawn from the US market at the request of the manufacturer, but intranasal oxytocin is still sold in countries outside the United States, such as Switzerland, Portugal, or Brazil. Recently, the use of oxytocin peptides in the treatment of autism spectrum disorders has been demonstrated. See WO2004 / 030524A2 and WO2008 / 042452A1, the disclosures of which are incorporated herein by reference. Autism spectrum disorder is becoming more and more prevalent in human population, and is typically recognized by certain behaviors and characteristics, such as impaired 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 show one or more of the above behaviors and characteristics to various degrees.Symptoms often observed in individuals with autism spectrum disorder are persistent deficits in social communication and social interaction, social anxiety, and repetitive behaviors, limited interests and activities.Other behaviors and characteristics observed in individuals with autism spectrum disorder also include avoidance of physical contact, generalized anxiety, monotony of vocalization or inability to regulate voice volume, inability to develop peer relationships, lack of shared enjoyment and interests, and lack of social or emotional reciprocity. Other disorders that exhibit social and communication deficits may 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 show symptoms similar to those exhibited in autism spectrum disorders). People with autism spectrum disorders tend to have communication deficits (e.g., responding inappropriately in conversations, misreading nonverbal interactions, or having difficulty building peer relationships appropriate for their age). In addition, people with autism spectrum disorders may be overly dependent on routines, be highly anxious and sensitive to changes in their environment, or focus intensely on inappropriate things (e.g., nanimate objects and / or narrow interests within a particular topic). Again, symptoms of people with autism spectrum disorder are widely variable and on a continuum, with some individuals exhibiting mild symptoms and others having severe symptoms. There are no pharmacological treatments available for the core deficits in social communication and social interaction, or the limited 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 the core symptoms of autism, particularly social and communication deficits and associated anxiety symptoms. Human clinical trials have demonstrated the effectiveness 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 have shown a wide variability in the response that people with autism spectrum disorder and related disorders have to be treated with oxytocin. Thus, there is a need for oxytocin peptide formulations that can provide more pronounced effects in the treatment of autism spectrum disorders 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 Summary of the Invention [Means for solving the problem]

[0006] Brief Summary of the Invention Methods and compositions are provided that include oxytocin peptides and magnesium ions for the treatment of autism spectrum disorders, related disorders, and symptoms of such disorders, including co-administration of oxytocin peptides and magnesium ions via craniofacial mucosal administration (e.g., intranasal administration).The methods and magnesium-containing oxytocin peptide formulations described herein provide enhanced efficacy in treating autism spectrum disorders compared to oxytocin alone.

[0007] In one aspect, the present invention provides a method for treating autism spectrum disorder, a disorder manifesting one or more symptoms associated with autism spectrum disorder, social and communication deficits, or anxiety, comprising administering to a subject in need thereof an effective dose of oxytocin peptide and magnesium ion, wherein co-administration of the oxytocin peptide and the magnesium ion results in a synergistic or enhanced effect. The oxytocin peptide and magnesium ion may be co-administered together or sequentially. In some embodiments, the oxytocin peptide is administered with the magnesium ion in the same unit dose or in a separate unit dose or formulation. In some embodiments, the oxytocin peptide and magnesium ion are administered sequentially. For example, the oxytocin peptide is administered a period of time after administration of the magnesium ion. In some embodiments, the subject is a human.

[0008] The oxytocin peptide and magnesium ions may be administered to a subject in need thereof via the same or different routes. In some embodiments, the oxytocin peptide is administered via craniofacial mucosal administration (e.g., nasal, buccal, sublingual, or ocular administration). In one embodiment, both the oxytocin peptide and magnesium ions are administered intranasally in the same formulation.

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

[0010] In one embodiment, a method for treating an autism spectrum disorder, a disorder manifesting one or more symptoms associated with an autism spectrum disorder, social and communication deficits, or anxiety comprising administering to a subject in need thereof an effective dose of an oxytocin peptide and magnesium ions, wherein co-administration of said oxytocin peptide and said magnesium ions results in a synergistic or enhanced effect, further comprising administering to said subject an effective dose of interleukin-6 (IL-6), wherein administration of IL-6 induces 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 dose 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 effective dose of magnesium ion administered is 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 method includes administering a magnesium salt (e.g., 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 method includes administering magnesium citrate or magnesium chloride in an amount providing 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 dose of oxytocin peptide and magnesium ions comprises about 0.5 μg to about 2000 μg or about 15 μg to about 120 μg (e.g., about 60 μg or about 66 μg) of oxytocin peptide administered in an aqueous solution containing about 0.11% to about 2.8% (preferably, about 1.1% to about 1.6%, e.g., about 1.36%) (w / v) magnesium.

[0012] In some embodiments, the present invention provides a method for reducing one or more symptoms associated with autism spectrum disorder. The symptoms treatable by the method include any social or communication deficit treatable by oxytocin peptides, such as eye contact deficits, social anxiety, generalized anxiety, accuracy in determining complex social cues, empathy, and communication abilities including expressive language functions.

[0013] In some embodiments, the present invention provides methods for the treatment of disorders that manifest one or more symptoms associated with an autism spectrum disorder, hi some embodiments, the disorder is 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 exhibited in autism spectrum disorders.

[0014] In one embodiment, the invention provides a method for treating autism spectrum disorder, comprising administering (e.g., intranasally) to a subject in need thereof an effective dose of oxytocin peptide and magnesium ion, wherein co-administration of the oxytocin peptide and the magnesium ion results in a synergistic or enhanced effect. In one embodiment, the invention provides a method for treating autism spectrum disorder, comprising administering (e.g., intranasally) to a subject in need thereof an effective dose of oxytocin peptide and magnesium ion, wherein the effective dose of the oxytocin peptide and the magnesium ion are administered intranasally in a liquid formulation, and the volume of the administered liquid formulation is from about 5 μL to about 1000 μL.

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

[0016] In one embodiment, a method for treating social and communication deficits is provided, comprising administering (e.g., intranasally) to a subject in need thereof an effective dose of oxytocin peptide and magnesium ion, wherein co-administration of the oxytocin peptide and the magnesium ion results in a synergistic or enhanced effect. In one embodiment, the present invention provides a method for treating social and communication deficits, comprising administering (e.g., intranasally) to a subject in need thereof an effective dose of oxytocin peptide and magnesium ion, wherein the effective dose of oxytocin peptide and magnesium ion are administered intranasally in a liquid formulation, and the volume of the administered liquid formulation is about 5 μL to about 1000 μL.

[0017] In one embodiment, a method for treating anxiety is provided, comprising administering (e.g., intranasally) to a subject in need thereof an effective dose of an oxytocin peptide and magnesium ions, wherein co-administration of the oxytocin peptide and the magnesium ions results in a synergistic or enhanced effect. In one embodiment, the present invention provides a method for treating anxiety, comprising administering to a subject in need thereof an effective dose of an oxytocin peptide and magnesium ions, wherein the effective dose of the oxytocin peptide and magnesium ions are administered intranasally in a liquid formulation, and the volume of the administered liquid formulation is from about 5 μL to about 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 the 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 the 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 dose of oxytocin peptide administered to the dose of magnesium ion administered is about 1:1 to about 1:1000. In some of these embodiments, the molar ratio of the dose of oxytocin peptide administered to the dose of magnesium ion administered is about 1:40 to about 1:40000. In some of these embodiments, the volume of the liquid formulation administered is about 50 μL to about 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 about 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 in a device for intranasal administration. In some of these embodiments, the device for intranasal administration is a nasal pump device. In some of these embodiments, the nasal pump device comprises a reservoir bottle attached to a pump actuator. In some of these embodiments, the pump actuator meters to deliver a specific volume of about 50 μL. In some of these embodiments, the nasal pump device comprises a reservoir bottle attached to an aerosolizer. 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 flow path, and (iii) a gamma-ray stable plastic material.

[0020] Further provided is the magnesium-containing oxytocin peptide formulation described herein for use in a subject in need thereof in a method for treating autism spectrum disorder, a disorder manifesting one or more symptoms associated with autism spectrum disorder, social and communication deficits, or anxiety.Also provided is the use of the magnesium-containing oxytocin peptide formulation described herein in the manufacture of a medicament for treating autism spectrum disorder, a disorder manifesting one or more symptoms associated with autism spectrum disorder, social and communication deficits, or anxiety.

[0021] Also provided is a kit that includes the magnesium-containing oxytocin peptide formulation described herein and suitable packaging material, which is included in a device for intranasal administration, such as a nasal pump device.The kit can further include instructions for administering the magnesium-containing oxytocin peptide formulation to a subject in need thereof 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) 1. A method for treating an autism spectrum disorder, a disorder manifesting one or more symptoms associated with an autism spectrum disorder, social and communication deficits, or anxiety, the method comprising administering to a subject in need thereof effective doses of an oxytocin peptide and magnesium ions, wherein co-administration of the oxytocin peptide and the magnesium ions results in a synergistic or enhanced effect. (Item 2) 2. The method of claim 1, wherein the oxytocin peptide is administered simultaneously with the magnesium ions. (Item 3) 2. The method of claim 1, wherein the oxytocin peptide is administered before or after administration of the magnesium ions. (Item 4) 4. The method according to any one of items 1 to 3, wherein the oxytocin peptide is administered via craniofacial mucosal administration. (Item 5) 5. The method of claim 4, wherein the oxytocin peptide is administered via intranasal administration. (Item 6) 6. The method of claim 5, wherein the oxytocin peptide and the magnesium ions are administered via intranasal administration. (Item 7) 7. The method according to any one of items 1 to 6, wherein the effective dose of the oxytocin peptide is from about 0.5 μg to about 2000 μg. (Item 8) 8. The method according to any one of items 1 to 7, wherein the effective dose of the magnesium ion is about 50 μg to about 68 mg. (Item 9) 9. The method according to any one of items 1 to 8, wherein the magnesium ions are provided using magnesium chloride and / or magnesium citrate. (Item 10) 2. The method according to claim 1, wherein the effective dose of the oxytocin peptide and the magnesium ion 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 11) 2. The method of claim 1, wherein the effective dose of the oxytocin peptide and the magnesium ion has an oxytocin:magnesium molar ratio of about 1:40 to about 1:40000. (Item 12) 12. The method according to any one of items 1 to 11, wherein the method is for treating an autism spectrum disorder. (Item 13) 12. The method according to any one of items 1 to 11, wherein the method is for treating a disorder exhibiting one or more symptoms associated with an autism spectrum disorder. (Item 14) The method according to item 13, wherein 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. 12. The method according to any one of items 1 to 11, wherein the method is for treating social and communication deficits. (Item 16) 12. The method according to any one of items 1 to 11, wherein the method is for treating anxiety. (Item 17) 17. The method according to any one of items 1 to 16, wherein the oxytocin peptide is human oxytocin (SEQ ID NO: 1). (Item 18) 1. A method for treating an autism spectrum disorder, a disorder manifesting one or more symptoms associated with an autism spectrum disorder, social and communication deficits, or anxiety, comprising the step of administering to a subject in need thereof an effective dose of an oxytocin peptide and magnesium ions, wherein the effective dose of the oxytocin peptide and magnesium ions are administered intranasally in a liquid formulation, and the volume of the administered liquid formulation is from about 5 μL to about 1000 μL. (Item 19) Item 19. The method according to item 18, wherein the effective dose of the oxytocin peptide is about 0.5 μg to about 2000 μg. (Item 20) Item 19. The method according to item 18, wherein the effective dose of the magnesium ion is about 50 μg to about 68 mg. (Item 21) 19. The method according to item 18, wherein the effective dose of the oxytocin peptide and the magnesium ion 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 method according to claim 18, wherein the effective dose of the oxytocin peptide and the magnesium ion has an oxytocin:magnesium molar ratio of about 1:40 to about 1:40000. 23. The method according to item 21 or 22, wherein the volume of the liquid formulation administered is about 50 μL to about 200 μL. (Item 24) 24. The method of claim 23, wherein the liquid formulation is administered using a metered dose nasal device in 1 to 4 units of about 50 μL / unit. (Item 25) 25. The method according to any one of items 18 to 24, wherein the method is for treating an autism spectrum disorder. (Item 26) 25. The method according to any one of items 18 to 24, wherein the method is for treating a disorder exhibiting one or more symptoms associated with an autism spectrum disorder. (Item 27) 28. The method according to claim 26, wherein 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. 25. The method according to any one of items 18 to 24, wherein the method is for treating social and communication deficits. (Item 29) 25. The method according to any one of items 18 to 24, wherein the method is for treating anxiety. . (Item 30) 30. The method according to any one of items 18 to 29, wherein the oxytocin peptide is human oxytocin (SEQ ID NO: 1). (Item 31) 19. The method of claim 18, wherein the liquid formulation is contained within a device for intranasal administration. (Item 32) 32. The method of claim 31, wherein the device for intranasal administration is a nasal pump apparatus. (Item 33) 33. The method of claim 32, wherein the nasal pump device comprises a reservoir bottle attached to a pump actuator. (Item 34) 34. The method of claim 33, wherein the pump actuator is metered to deliver a specific volume of about 50 μL. (Item 35) Item 33. The method of item 32, wherein the nasal pump device comprises a storage bottle attached to an aerosolizer. (Item 36) The nasal pump device comprises: (i) a filter to prevent backflow; (ii) a metal-free flow path; and (iii) Gamma-ray-stable plastic materials 36. The method according to any one of items 32 to 35, comprising one or more of the following: (Item 37) 1. A composition comprising an oxytocin peptide and magnesium ions, wherein said oxytocin peptide and said magnesium ions are present in amounts that provide a synergistic or enhanced effect when used to treat anxiety. (Item 38) 38. The composition of claim 37, wherein the oxytocin peptide is human oxytocin (SEQ ID NO: 1). (Item 39) 38. The composition of claim 37, wherein the composition is a liquid formulation comprising about 0.01 mg / mL to about 16 mg / mL of the oxytocin peptide. (Item 40) 38. The composition according to item 37, wherein the composition is a liquid formulation comprising the magnesium in an amount providing from about 3 mg / mL to about 30 mg / mL of magnesium. (Item 41) Item 38. The composition according to item 37, wherein the oxytocin peptide and the magnesium ion have a molar ratio of about 1:40 to about 1:40000. (Item 42) Item 42. The composition according to item 41, wherein the molar ratio is from about 1:40 to about 1:800. (Item 43) Item 42. The composition according to item 41, wherein the molar ratio is about 1:800 to about 1:40000. (Item 44) 44. The composition according to any one of items 37 to 43, further comprising a device for administration to the craniofacial mucosa. (Item 45) 45. The composition of claim 44, wherein the oxytocin peptide and the magnesium ions are contained in a device for administration to the craniofacial mucosa. (Item 46) 46. ​​The composition of claim 45, wherein the device is for intranasal administration. [Brief description of the drawings]

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

[0023] [Diagram 2] FIG. 2 shows the effects of saline, oxytocin, magnesium citrate and a combination of oxytocin, and magnesium citrate in a rat model of anxiety.

[0024] [Diagram 3] 3A and 3B show the effects of magnesium citrate, oxytocin, and the combination of magnesium citrate and oxytocin in the elevated plus maze rat model of anxiety. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0025] Detailed Description of the Invention The present invention provides, inter alia, a method for treating autism spectrum disorder, a disorder manifesting one or more symptoms associated with autism spectrum disorder, social and communication deficits, or anxiety in a subject in need thereof by craniofacial administration (e.g., intranasal administration) of oxytocin peptide and magnesium ion, or a magnesium-containing oxytocin peptide formulation as described herein.The oxytocin peptide and magnesium ion are administered in an effective dose that produces a synergistic or enhanced effect compared to administration of oxytocin peptide alone.

[0026] (definition) As used herein, "oxytocin peptide" refers to a substance that has biological activity related to natural oxytocin. The oxytocin peptide can be a naturally occurring endogenous peptide, a fragment thereof, an analog, or a derivative thereof. The oxytocin peptide can also be a non-endogenous peptide, a fragment thereof, an analog, or a derivative thereof. In one embodiment, the oxytocin peptide is human oxytocin. In another embodiment, the oxytocin peptide can be an analog or a derivative of human oxytocin.

[0027] As used herein, "analog" or "derivative" refers to any peptide similar to naturally occurring oxytocin, in which one or more amino acids are substituted, deleted or inserted.This term also refers to 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, this term encompasses all peptides that exhibit oxytocin activity, but may have different potency or pharmacological profile if desired.

[0028] As used herein, unless otherwise specified, the term "treatment" or "treating" refers to an approach to obtain beneficial or desired results (e.g., clinical results).With respect to autism spectrum disorder and related disorders, beneficial or desired results include, but are not limited to, the alleviation of symptoms and / or the severity of symptoms, such as, but not limited to, social and / or communication deficits and / or repetitive behaviors and / or anxiety.Social and communication deficits may include, but are not limited to, impaired communication skills and / or social interaction, lack of eye contact, and / or inability to form and / or maintain social relationships.

[0029] "Synergy", "synergism" or "synergy" refers to the combined action of two or more compounds in such a manner that one complements or enhances the action of the other, resulting in an effect that is greater than would be predicted or expected by adding up the effects of the two or more compounds when administered individually at a given dose. A "synergistic effect" is considered to be achieved when the combined use of two or more drugs results in a greater overall effect (e.g., improvement in social and communication deficits and / or reduction in anxiety) than would be predicted or expected by adding up the effects of either of them individually at equal doses. A "synergistic effect" is also considered to be achieved when the combined use of two or more drugs results in a faster onset of effect and / or a longer lasting effect than would occur following administration of each drug alone at equal doses.

[0030] "Cranio-facial mucosal administration" refers to delivery to the mucosal surfaces of the nose, nasal passages, and nasal cavity; mucosal surfaces of the oral cavity including the gingiva (gums), floor of the mouth, lips, tongue, sublingual oral surfaces (including the lingual frenulum and floor of the mouth), and mucosal surfaces of or around the eye (including the conjunctiva, lacrimal gland, nasolacrimal duct) and the upper or lower eyelids and mucous membranes of the eye.

[0031] "Intranasal administration" or "administered intranasally" refers to delivery by spray, drops, powder, gel, film, inhalant or other means to the nose, nasal passages or nasal cavity.

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

[0033] "Subject" or "patient" as used herein refers to a mammal, including, but not limited to, a human. Mammals include, but are not limited to, farm animals (e.g., cows), sport 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" include plural referents unless otherwise indicated. Additionally, as used herein, the term "comprising" and its cognates are used in their inclusive sense; that is, they are equivalent to the term "including" and its corresponding cognates.

[0035] When a range of values ​​is provided, each intervening value between the upper and lower limits of the range and any other stated or intervening value within the stated range is intended to be encompassed within the scope of the disclosure. For example, if a range of 1 μg to 8 μg is provided, 2 μg, 3 μg, 4 μg, 5 μg, 6 μg, and 7 μg, as well as ranges of values ​​of 1 μg or more and 8 μg or less are also intended to be expressly disclosed. If a range of 10 to 14% is provided, 10%, 11%, 12%, 13%, and 14% are also intended to be expressly disclosed. In addition, each narrower range within a stated range between any stated or intervening value and any other stated or intervening value within the stated range is encompassed within the scope of the disclosure. The upper and lower limits of these narrower ranges may independently be included or excluded from the ranges, and each range in which one or both limits are included in the narrower range, or in which neither limit is included in the narrower range, is also included in the disclosure subject to any specifically excluded limit in that stated range. When a stated range includes one or both limits, ranges excluding either or both of those included limits are 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 cyclic peptide hormone of nine amino acids with two cysteine ​​residues forming a disulfide bridge 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. For example, see U.S. Patent No. 2,938,891 and U.S. Patent No. 3,076,797. In addition, oxytocin is commercially available. A variety of peptide analogs and peptide derivatives are available, and others may be contemplated for use in the present invention and may be produced according to known methods and tested for biological activity. Oxytocin analogs may include, but are not limited to, 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 related series of compounds containing the ortho-trifluor-ethoxyphenylacetyl core, such as L-374,943.Other exemplary oxytocin analogs include 4-threonine-1-hydroxy-deaminooxytocin, 9-deaminooxytocin, analogs of oxytocin containing glycine residues in place of glycinamide residues, (2,4-diisoleucine)-oxytocin, analogs of oxytocin that have natriuretic and diuretic activity, deaminooxytocin analogs; long-acting oxytocin analogs, 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-carba-oxytocin (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-deamino-oxytocin (residues 1 and 6). and desamino-oxytocin analogs, in which the disulfide bridge between the oxytocin residues is replaced by a thioether, and desamino-oxytocin analogs, in which the disulfide bond is replaced by a diselenide, ditelluride, telluroseleno, tellurosulfide or selenosulfide bond (e.g., peptide analogs of oxytocin described in PCT patent application WO2011 / 120,071, which is incorporated herein by reference). Peptides for use in the present invention may be peptides that are obtainable by partial substitution, addition or deletion of amino acids within a naturally occurring or naturally occurring peptide sequence.The peptides may be chemically modified, for example, by amidation (-NH2) of the carboxyl terminus, use of D-amino acids in the peptide, 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 native 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. The present disclosure also encompasses other known oxytocin analogs, such as the peptidic 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 which are incorporated herein by reference. In some embodiments, the oxytocin analog is a peptide oxytocin receptor agonist described in Wisniewski et al., J. A compound 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 merotocin).

[0038] In some embodiments, oxytocin or an oxytocin analog is isotopically labeled by having one or more atoms replaced with an isotope having a different atomic mass. Examples of isotopes that can be incorporated into the disclosed compounds include isotopes of hydrogen (e.g., 2 H and 3 H), isotopes of carbon (e.g. 13 C and 14 C), isotopes of nitrogen (e.g. 15N), isotopes of oxygen (e.g. 18 O and 17 O), phosphorus isotopes (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 may be administered to a subject or other subjects according to conventional techniques and then detected to provide useful diagnostic and / or therapeutic management data. Furthermore, isotopically labeled compounds may be administered to a subject or other subjects in need thereof to provide therapeutically beneficial absorption, distribution, metabolism and / or excretion profiles. All isotopic variations of oxytocin peptides, e.g., human oxytocin or analogs or derivatives thereof, whether radioactive or not, are contemplated.

[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] An "International Unit" (IU, UI or IE) is an internationally accepted unit of activity used to quantify vitamins, hormones and vaccines. An International Unit defines the amount of a substance that provides a unit of activity measured using a defined biological assay to standardize preparations from multiple source materials. Similarly, a USP unit is a defined dosage unit established by the United States Pharmacopeia in cooperation with the Food and Drug Administration to ensure the identity, strength, quality, purity and consistency of drug products. Generally, the USP unit is equal to the International Unit due to an attempt at harmonization. By convention, in the case of oxytocin, one unit of activity is usually defined as being equal to approximately 2 micrograms of synthetic oxytocin peptide; or 1 mg is equal to 500 units (Stedman's Medical Dictionary). Thus, as used herein, one "IU" or one "International Unit" of oxytocin peptide is the amount of oxytocin peptide that has the same biological activity or produces the same level of biological action (e.g., contractile response of rat uterine strips) as approximately 2 micrograms of synthetic peptide. Less active analogs would require more substance to achieve the same level of biological effect. Measurement of drug potency is well known to those skilled in the art and can include either in vitro or in vivo assays using synthetic oxytocin as 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 formulation) In the method of the present invention for treating autism spectrum disorder, one or more symptoms associated with autism spectrum disorder, social and communication deficits, or anxiety, which comprises administering an effective dose of oxytocin peptide and magnesium ion to a subject in need thereof, said oxytocin peptide and said magnesium ion can be administered in a magnesium-containing oxytocin peptide preparation or composition.In one embodiment, said magnesium-containing oxytocin peptide preparation or composition comprises oxytocin peptide and magnesium ion in an amount that produces synergistic or enhanced effect when used in treating autism spectrum disorder, one or more symptoms associated with autism spectrum disorder, social and communication deficits, or anxiety.

[0042] The relative ratio of oxytocin peptide and magnesium ion in magnesium-containing oxytocin peptide formulations is important in achieving optimal synergistic or enhanced effect.The optimal amount of oxytocin peptide and magnesium ion may depend on other factors such as specific disorder or symptoms, the type of synergistic or enhanced effect desired, and route of administration.For example, the amount of magnesium may be important to make the effect occur faster; the amount of oxytocin may be important to make the effect last longer, and the relative ratio of oxytocin and magnesium may be important to achieve the greatest improvement in social function, reduction of social and communication deficits, and / or reduction of anxiety.

[0043] In some embodiments, the magnesium-containing oxytocin peptide formulation or composition is a liquid formulation 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 formulation 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 formulation 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. That is, the amount of oxytocin peptide in the liquid formulation is anywhere in the range of about 0.01 to 16 mg / mL, with the lower limit being less than the upper limit.In some embodiments, the magnesium-containing oxytocin peptide formulation or composition comprises from about 0.01 mg / mL to about 12 mg / mL, from about 0.05 mg / mL to about 16 mg / mL, from about 0.1 mg / mL to about 12 mg / mL, from about 0.1 mg / mL to about 8 mg / mL, from about 0.1 mg / mL to about 4 mg / mL, from about 0.1 mg / mL to about 2 mg / mL, from about 0.1 mg / mL to about 1.6 mg / mL, from about 0.1 mg / mL to about 1.2 mg / mL, from about 0.1 mg / mL to about 1. mg / mL, approximately 0.1 mg / mL to approximately 0.8 mg / mL, approximately 0.1 mg / mL to approximately 0.4 mg / mL, approximately 0.1 mg / mL to approximately 0.3 mg / mL, approximately 0.2 mg / mL to approximately 16 mg / mL, approximately 0.2 mg / mL to approximately 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 In a preferred embodiment, the magnesium-containing oxytocin peptide formulation or composition comprises about 0.1 mg / mL to about 2 mg / mL, about 0.15 mg / mL to about 1.5 mg / mL, or about 0.2 mg / mL to about 1.2 mg / mL of oxytocin peptide.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 about 5 IU / mL to about 8000 IU / mL of oxytocin peptide. 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 about (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 anywhere in the range of about 5 to 8000 IU / mL, with the lower limit being less than the upper limit.In some embodiments, the magnesium-containing oxytocin peptide formulation or composition comprises from about 500 IU / mL to about 6000 IU / mL, from about 25 IU / mL to about 8000 IU / mL, from about 50 IU / mL to about 6000 IU / mL, from about 50 IU / mL to about 4000 IU / mL, from about 50 IU / mL to about 2000 IU / mL, from about 50 IU / mL to about 1000 IU / mL, from about 50 IU / mL to about 800 IU / mL, from about 50 IU / mL to about 600 IU / mL, from about 50 IU / mL to about 5 ... 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 about 150 IU / mL to about 1500 IU / mL, about 150 IU / mL to about 500 IU / mL, about 150 IU / mL to about 250 IU / mL, about 250 IU / mL to about 8000 IU / mL, about 250 IU / mL to about 5000 IU / mL, about 250 IU / mL to about 2500 IU / mL, about 250 IU / mL to about 500 IU / mL, about 500 IU / mL to about 8000 IU / mL, about 500 IU / mL to about 5000 IU / mL, or about 500 IU / mL to about 2500 IU / mL of oxytocin peptide.In a preferred embodiment, the magnesium-containing oxytocin peptide formulation or composition comprises about 50 IU / mL to about 1000 IU / mL, about 75 IU / mL to about 750 IU / mL, or about 100 IU / mL to about 600 IU / mL of oxytocin peptide. 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 formulations is expressed as a percentage by weight (w / v) (Magnesium or Mg per 100 mL of solution). 2+ grams of magnesium or Mg per milliliter of solution 2+ (milligrams of magnesium or Mg per liter of solution) or molar concentration ( 2+ "M" defined as moles of magnesium or Mg per liter of solution 2+ It can also be expressed as "mM", which is defined as millimoles of

[0046] In some embodiments, the magnesium-containing oxytocin peptide formulation or composition contains about 1 mg / mL to about 30 mg / mL of magnesium or magnesium ions (Mg 2+). 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 anywhere in the range of about 1 to 30 mg / mL, with the lower limit being less than the upper limit. In some embodiments, the magnesium-containing oxytocin peptide formulation or composition contains 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+ In some embodiments, the magnesium-containing oxytocin peptide formulation or composition contains about 50 mM to about 1500 mM magnesium or magnesium ions (Mg 2+In some embodiments, the amount of magnesium or magnesium ions in the liquid formulation is greater than about (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 about (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 anywhere in the range of about 50 to 1500 mM, with the lower limit being less than the upper limit. 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 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+ 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 comprising:

[0047] Any magnesium salt (e.g., water-soluble magnesium salt) can be used to provide magnesium ions in magnesium-containing oxytocin peptide formulations. The magnesium salt used in the magnesium-containing oxytocin peptide formulations can be selected based on many factors, such as the amount of free magnesium ions that can be delivered when the formulation is administered, the solubility of the magnesium salt in the liquid formulation medium, 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 provide a synergistic or enhanced effect with the oxytocin peptide. When selecting a magnesium salt, other factors can also be considered, such as compatibility with other substances in the formulation and the ability of the counterion to perform other functions in the formulation. For example, magnesium citrate is sufficiently soluble in aqueous solution to provide the desired amount of magnesium or the desired magnesium ion concentration; the citrate salt can be pharma- ceutically acceptable; the citrate salt can be part of a buffer; and the magnesium citrate salt can add a favorable flavor to the formulation. The magnesium ion in magnesium-containing oxytocin peptide formulation can be provided by using one or more magnesium salts.The magnesium salt in magnesium-containing oxytocin peptide formulation can be the magnesium salt that is first used in preparing magnesium-containing oxytocin peptide formulation, or can be formed in situ during the preparation of magnesium-containing oxytocin peptide formulation.For example, magnesium chloride can be first used in the preparation of formulation; magnesium citrate can be formed in situ when citric acid is added to the formulation.In such a case, the magnesium ion in magnesium-containing oxytocin peptide formulation is provided by both magnesium chloride and magnesium citrate.

[0048] In some embodiments, the magnesium-containing oxytocin peptide formulation 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 maltate, magnesium taurate, magnesium gluconate, magnesium succinate, and magnesium pyrophosphate. In some embodiments, the magnesium-containing oxytocin peptide formulation or composition is a liquid formulation comprising a magnesium salt (e.g., magnesium citrate or magnesium chloride) in an amount providing from about 1 mg / mL to about 30 mg / mL of magnesium. In some embodiments, the composition comprises from about 1 mg / mL to about 30 mg / mL of magnesium ion (Mg 2+ In some embodiments, the magnesium-containing oxytocin peptide formulation or composition contains about 1 mg / mL to about 30 mg / mL of magnesium or magnesium ions (Mg 2+ In some embodiments, the composition comprises one or more magnesium salts in an amount to provide 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 comprises 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+In some embodiments, the magnesium-containing oxytocin peptide formulation or composition is a liquid formulation comprising 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 a magnesium salt (e.g., magnesium citrate or magnesium chloride) in an amount that provides a therapeutic effect. In some embodiments, the magnesium-containing oxytocin peptide formulation or composition comprises 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+ A liquid formulation containing 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 one or more magnesium salts (e.g., magnesium citrate and / or magnesium chloride) in an amount that provides a therapeutic effect. In some embodiments, the magnesium-containing oxytocin peptide formulation or composition is 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+ and one or more magnesium salts (e.g., magnesium citrate and / or magnesium chloride).

[0049] The relative amounts of oxytocin peptide and magnesium ion in the magnesium-containing oxytocin peptide formulation or composition detailed herein may be defined by weight ratio or molar ratio. The weight ratio of the amount of oxytocin peptide to the amount of magnesium or magnesium ion in the 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 each 1 mg of oxytocin peptide present in the formulation or composition, the amount of magnesium or magnesium ion present in the formulation or composition is about 40 mg. The molar ratio of the amount of oxytocin peptide to the amount of magnesium or magnesium ion in the 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 each 1 μmol of oxytocin peptide present in the formulation or composition, the amount of magnesium or magnesium ion present in the formulation or composition is about 1600 μmol.

[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 formulation or composition is anywhere in the range of 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 from about 1:2 to about 1:1000, from about 1:2 to about 1:800, from about 1:2 to about 1:500, from about 1:2 to about 1:250, from about 1:2 to about 1:150, from about 1:2 to about 1:100, from about 1:2 to about 1:80, from about 1:2 to about 1:60, from about 1:2 to about 1:50, from about 1:2 to about 1:40, from about 1:2 to about 1:30, from about 1:2 to about 1:20, from about 1:2 to about 1:10, from about 1:2 to about 1:5, from about 1:5 to about 1:1000, from about 1:5 to about 1:800, from about 1:5 to about 1:500, from about 1:5 to about 1:200, 5 to about 1:100, about 1:5 to about 1:80, about 1:5 to about 1:60, about 1:5 to about 1:50, about 1:5 to about 1:40, about 1:5 to about 1:30, about 1:5 to about 1:20, about 1:5 to about 1:10, about 1:10 to about 1:1000, about 1:10 to about 1:800, about 1:10 to about 1:500, about 1:10 to about 1:200, about 1:10 to about 1:100, about 1:10 to about 1:80, about 1:10 to about 1:60, about 1:10 to about 1:50, about 1:10 to about 1:40, about 1:10 to about 1:30, about 1:10 to about 1:20, about 1:20 to about 1:1000,About 1:20 to about 1:800, about 1:20 to about 1:500, about 1:20 to about 1:200, about 1:20 to about 1:100, about 1:20 to about 1:80, about 1:20 to about 1:70, about 1:20 to about 1:60, about 1:20 to about 1:50, about 1:20 to about 1:40, about 1:20 to about 1:30, about 1:30 to about 1:1000, about 1:30 to about 1:800, about 1:30 to about 1:500, about 1:30 to about 1:200, about 1:30 to about 1:100, about 1:30 to about 1:80, about 1:30 to about 1:70, about 1:30 to about 1:60, about 1:30 to about 1:50, about 1:30 to about 1:40, about 1:35 to about 1:45, about 1:40 to about 1:1000, about 1:40 to about 1:800, about 1:40 to about 1:500, about 1:40 to about 1:200, about 1:40 to about 1:100, about 1:40 to about 1:80, about 1:40 to about 1:70, about 1:40 to about 1:60, about 1:40 to about 1:50, about 1:50 to about 1:1000, about 1:50 ~ about 1:800, about 1:50 to about 1:500, about 1:50 to about 1:200, about 1:50 to about 1:100, about 1:50 to about 1:90, about 1:50 to about 1:80, about 1:50 to about 1:70, about 1:50 to about 1:60, about 1:60 to about 1:1000, about 1:60 to about 1:800, about 1:60 to about 1:50, about 1:60 to about 1:200, about 1:60 to about 1:100, about 1:60 to about 1:90, about 1:60 to about 1:80, about 1:60 to about 1:70, having an OT / Mg(w) ratio of about 1:80 to about 1:1000, about 1:80 to about 1:800, about 1:80 to about 1:500, about 1:80 to about 1:200, about 1:80 to about 1:100, about 1:100 to about 1:1000, about 1:100 to about 1:800, about 1:100 to about 1:500, about 1:100 to about 1:200, about 1:200 to about 1:1000, about 1:200 to about 1:800, about 1:200 to about 1:500, or about 1:500 to about 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 the 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 formulation or composition is greater than about (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 formulation or composition is anywhere in the range of about 1:40 to 1:40000, with the upper limit being greater than the lower limit. In some embodiments, the formulation or composition has an OT / Mg(m) ratio of about 1:80 to about 1:8000. In some preferred embodiments, the formulation or composition has 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 formulation or composition has 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 1:2 ... :80 to about 1:1600, about 1:80 to about 1:1200, about 1:80 to about 1:800, about 1:80 to about 1:400, about 1:80 to about 1:200, about 1:175 to about 1:40000, about 1:175 to about 1:30000, about 1:175 to about 1:20000, about 1:175 to about 1:10000, about 1:175 to about 1:5000,About 1:175 to about 1:3000, about 1:175 to about 1:2400, about 1:175 to about 1:2000, about 1:175 to about 1:1700, about 1:175 to about 1:1600, about 1:175 to about 1:1200, about 1:175 to about 1:1100, about 1:175 to about 1:800, about 1:17 5 to about 1:560, about 1:175 to about 1:400, about 1:175 to about 1:280, about 1:200 to about 1:40000, about 1:200 to about 1:30000, about 1:200 to about 1:20000, about 1:200 to about 1:10000, about 1:200 to about 1:5000, about 1:200 to about 1 :3000, 1:200~1:2400, 1:200~1:2000, 1:200~1:1600, 1:200~1:1200, 1:200~1:800, 1:200~1:400, 1:280~1:40000, 1:280~1:3000 0, about 1:280 to about 1:20000, about 1:280 to about 1:10000, about 1:280 to about 1:5000, about 1:280 to about 1:3000, about 1:280 to about 1:2400, about 1:280 to about 1:2000, about 1:280 to about 1:1700, about 1:280 to about 1:1600, about 1:280~1:1200, 1:280~1:1100, 1:280~1:800, 1:280~1:560, 1:280~1:400, 1:400~1:40000, 1:400~1:30000, 1:40~1:20000, 1:40 0~1:8000, 1:400~1:4000, 1:400~1:3000, 1:400~1:2400, 1:400~1:2000, 1:400~1:1600, 1:400~1:1200, 1:400~1:800, 1:560~1:4 0000, about 1:560 to about 1:30000, about 1:560 to about 1:20000, about 1:560 to about 1:8000, about 1:560 to about 1:4000, about 1:560 to about 1:3000, about 1:560 to about 1:2400, about 1:560 to about 1:2000, about 1:560 to about 1:170 0, about 1:560 to about 1:1600, about 1:560 to about 1:1200, about 1:560 to about 1:1100, about 1:560 to about 1:800, about 1:800 to about 1:40000, about 1:800 to about 1:30000, about 1:800 to about 1:20000, about 1:800 to about 1:10000,About 1:800 to about 1:5000, about 1:800 to about 1:3000, about 1:800 to about 1:2400, about 1:800 to about 1:2000, about 1:800 to about 1:1600, about 1:800 to about 1:1200, about 1:1100 to about 1:40000, about 1:1100 to about 1:30000, About 1:1100~about 1:20000, about 1:1100~about 1:10000, about 1:1100~about 1:5000, about 1:1100~about 1:4000, about 1:1100~about 1:3000, about 1:1100~about 1:2400, about 1:1100~about 1:2000, about 1:1100~about 1: 1700, approx. 1:1100~approx. 1:1600, approx. 1:1200~approx. 1:40000, approx. 1:1200~approx. 1:30000, approx. 1:1200~approx. 1:20000, approx. 1:1200~approx. 1:10000, approx. 1:1200~approx. 1:5000, approx. 1:1200~approx. 1:4000, approx. 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, about 1:1600 to about 1:10000, about 1:1600 to about 1:5000, about 1:1600 to about 1:3000, about 1:1600 to about 1:2400, about 1:1600 to about 1:2000, about 1:1700 to about 1:40000, about 1:1700 to about 1:30000, about 1:170 0~approx. 1:20000, approx. 1:1700~approx. 1:10000, approx. 1:1700~approx. 1:5000, approx. 1:1700~approx. 1:3000, approx. 1:1700~approx. 1:2400, approx. 1:1700~approx. 1:2000, approx. 1:2000~approx. 1:40000, approx. 1:2000~approx. 1:30000 , about 1:2000 to about 1:20000, about 1:2000 to about 1:10000, about 1:2000 to about 1:5000, about 1:2000 to about 1:4000, about 1:2000 to about 1:3000, about 1:2000 to about 1:2400, about 1:2400 to about 1:40000, about 1:2400 to About 1:30000, about 1:2400 to about 1:20000, about 1:2400 to about 1:10000, about 1:2400 to about 1:5000, about 1:2400 to about 1:4000, about 1:2400 to about 1:3000, about 1:3000 to about 1:40000, about 1:3000 to about 1:30000,having an OT / Mg(m) ratio of about 1:3000 to about 1:20000, about 1:3000 to about 1:10000, about 1:3000 to about 1:4000, about 1:4000 to about 1:40000, about 1:4000 to about 1:30000, about 1:4000 to about 1:20000, about 1:4000 to about 1:10000, about 1:8000 to about 1:40000, about 1:8000 to about 1:30000, about 1:8000 to about 1:20000, or about 1:10000 to about 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, the magnesium-containing oxytocin peptide formulation or composition comprising oxytocin peptide and magnesium ion further comprises one or more pharma- ceutically acceptable carriers (thus constituting a pharmaceutical composition) and optionally other components (e.g., excipients, vehicles, emulsifiers, stabilizers, preservatives, buffers, and / or other additives that may enhance stability, delivery, absorption, half-life, efficacy, pharmacokinetics and / or pharmacodynamics, reduce adverse side effects, or provide other benefits for pharmaceutical use).Exemplary excipients include solubilizers, surfactants, and chelating agents. For example, the formulation may include 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] Liquid carriers include, but are not limited to, water, saline, aqueous dextrose, and glycols, especially for solutions (when isotonic). The carriers can also be selected from a variety of oils, including those of petroleum, animal, vegetable, or synthetic origin (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, flour, chalk, silica gel, magnesium stearate, sodium stearate, glycerol monostearate, sodium chloride, skimmed milk powder, glycerol, propylene glycol, water, ethanol, and the like. The compositions can be subjected to conventional pharmaceutical processes such as sterilization, and can contain conventional pharmaceutical additives (e.g., preservatives, stabilizing agents, reducing agents, antioxidants, chelating agents, wetting agents, emulsifying agents, dispersing agents, jelling agents, salts for adjusting osmotic pressure, buffers, and the like). 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 a preferred embodiment, the pH of the formulation / composition is about 4.5.

[0054] In some embodiments, the magnesium-containing oxytocin peptide formulation or composition may further comprise one or more mucosal delivery enhancers selected from (A)-(K): (A) solubilizing agents; (B) charge modifying agents; (C) pH adjusting agents; (D) degradative enzyme inhibitors; (E) mucolytic or mucus clearing agents; (F) ciliostatic agents; (G) membrane permeability enhancers; (H) modulators of epithelial junctional physiology (e.g., nitric oxide (NO) stimulators, chitosan and chitosan derivatives); (I) vasodilators; (J) selective transport enhancers; and (K) stabilizing delivery vehicles, carriers, carriers or complexing species that are effectively combined with, associated with, contained in, encapsulated or bound to the oxytocin peptide to stabilize the active agent for enhanced mucosal delivery. The membrane permeation enhancers in group (G) can 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 amphipathic molecules, (viii) small molecule hydrophobic permeation enhancers; (ix) sodium or salicylic acid derivatives; (x) glycerol esters of acetoacetic acid, (xi) cyclodextrins or beta-cyclodextrin derivatives, (xii) medium chain fatty acids, (xiii) chelating agents, (xiv) amino acids or salts thereof, (xv) N-acetyl amino acids or salts thereof, (xvi) enzymes degradative to selected membrane components, (xvii) inhibitors of fatty acid synthesis, (xviii) inhibitors of cholesterol synthesis; or (xiv) any combination of the membrane permeation enhancers of (i)-(xviii). In various embodiments of the invention, the oxytocin peptides may be combined with one, two, three, four or more of the mucosal delivery enhancing agents listed in (A) through (K). These mucosal delivery enhancing agents, alone or together, may be admixed with the oxytocin peptides or otherwise combined therewith in a pharma- ceutically acceptable formulation or delivery vehicle.The magnesium-containing oxytocin peptide formulations or compositions described herein may increase the bioavailability of the oxytocin peptide after it is delivered to a mucosal surface (e.g., the mucosal surface of the nasal cavity) of a mammalian subject.

[0055] The list of carriers and excipients discussed herein is by no means exhaustive, and one of skill in the art can select carriers and excipients from the GRAS (generally regarded as safe) list of chemicals approved in pharmaceuticals and those currently approved by the U.S. Food and Drug Administration in topical and parenteral formulations, as well as those that will be approved 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 formulation or composition comprising an oxytocin peptide and magnesium ions, wherein the oxytocin peptide and magnesium ions are present in amounts that provide a synergistic 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, sodium monohydrogen phosphate, sorbitol, and water. In some embodiments, the magnesium-containing oxytocin peptide formulation or composition further comprises chlorobutanol, acetic acid, and water.

[0057] In some embodiments, the magnesium-containing oxytocin peptide formulation or composition comprising 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 a chitosan-containing excipient. In some embodiments, chitosan glutamate may be preferred for nasal delivery due to its superior 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 comprising insulin and reduced glutathione and may also be useful as an excipient in the magnesium-containing oxytocin peptide formulation or composition described herein.

[0058] In some embodiments, the magnesium-containing oxytocin peptide formulation or composition comprising oxytocin peptide and magnesium ions further comprises one or more gelling agents so that the oxytocin peptide formulation forms a gel in the nasal cavity, thus enhancing the nasal absorption of the oxytocin peptide. The gelling system useful in the formulations and methods described herein can include any known gelling system, such as chemically reactive pectin-based gelling systems (e.g., PecSys™, Archimedes Pharma) and thermoresponsive polymer gelling systems (e.g., Pluronic® F127, BASF). PecSys™ is a low-viscosity pectin-based aqueous solution delivered as a fine mist that gels when each droplet comes into contact with calcium ions in the nasal mucosa. Other low methoxy pectins can also be used, for example, at about 1% concentration. Pluronic® F127 comprises an ethylene oxide / propylene oxide block copolymer. The gelling temperature varies depending on the ratio of components and the amount of copolymer used in the final formulation. Gelling in the human nasal cavity has been demonstrated for approximately 18-20% wt / vol Pluronic® F127, for example, used in vitamin B12 gel supplements (EnerB, Nature's Bounty, NY) and gelling sumatriptan with 18% wt / vol Pluronic® F127 and 0.3% wt / vol Carbopol (anionic bioadhesive polymer C934P). The ratio and concentration of monomers can be adjusted for the intended oxytocin formulation to ensure gelation at 25-37°C, near the standard temperature of 34°C in the nasal cavity. If the gelling temperature is below 25°C, the formulation may gel at room temperature; if the gelling 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 include a mucoadhesive material, such as Carbopol. The addition of a mucoadhesive agent, for example up to 0.5% Carbopol, can further reduce the gelling temperature.

[0059] In some embodiments, the magnesium-containing oxytocin peptide formulation or composition comprising oxytocin peptide and magnesium ions further comprises a surfactant, such as a non-ionic surfactant (e.g., polysorbate-80) and one or more buffers, stabilizers, or tonicifiers. In some embodiments, the magnesium-containing oxytocin peptide formulation or composition further comprises a propellant. The pH of the nasal spray solution is optionally about pH 3.0-8.5, although if desired, the pH is adjusted to optimize delivery of the charged macromolecular species (e.g., therapeutic protein or peptide) in a substantially non-ionized state. The pharmaceutical solvent used may also be a slightly acidic aqueous buffer (pH 3-6). Suitable buffers for use in these compositions are as described above or otherwise known in the art. Other components may be added to enhance or maintain chemical stability, including preservatives, surfactants, dispersants, or gases. Suitable preservatives include, but are not limited to, phenol, methylparaben, paraben, m-cresol, thiomersal, benzalkonium chloride, and the like. Suitable surfactants include, but are not limited to, oleic acid, sorbitan trioleate, polysorbates, lecithin, phosphotidylcholine, and various long-chain diglycerides and phospholipids.Suitable dispersants include, but are not limited to, ethylenediaminetetraacetic acid (EDTA), and the like.Suitable gases include, but are not limited to, nitrogen, helium, chlorofluorocarbons (CFCs), hydrofluorocarbons (HFCs), carbon dioxide, air, and the like.Suitable stabilizers and tonicifying 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 formulation or composition comprising an oxytocin peptide and magnesium ions further comprises an agent capable of upregulating oxytocin receptor expression (eg, IL-6).

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

[0062] Absorption enhancers may be included in the formulation to enhance delivery and / or absorption of oxytocin peptide and magnesium ion into or across mucosal surfaces. These enhancers may increase the release or solubility (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., as measured at the delivery site) of the composition. Thus, enhanced mucosal delivery may occur by any of a variety of mechanisms, such as by increasing the diffusion, transport, persistence or stability of the oxytocin peptide, increasing membrane fluidity, modulating the availability or action of calcium and other ions that control intracellular or paracellular permeability, solubilizing mucosal components (e.g., lipids), altering non-protein and protein sulfhydryl levels in mucosal tissues, increasing water flux across mucosal surfaces, modulating epithelial junctional physiology, decreasing the viscosity of the mucus covering the mucosal epithelium, decreasing the rate of mucociliary clearance, and other mechanisms.

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

[0064] All of the peptides described and / or contemplated herein may be prepared by chemical synthesis using automated or manual solid phase synthesis techniques that are well known in the art. The peptides may also be prepared using recombinant molecular methods known in the art.

[0065] (Delivery system) The magnesium-containing oxytocin peptide formulation or composition may be adapted for craniofacial mucosal administration (e.g., nasal, buccal, sublingual or ocular administration). In some embodiments, the composition may further comprise a device for mucosal delivery. In some embodiments, the composition is adapted for buccal and / or sublingual mucosal delivery, and the composition may further comprise a device for buccal and / or sublingual mucosal administration (e.g., unit dose container, pump spray, dropper, squeeze bottle, preservative-free airless spray, nebulizer, dose inhaler and pressurized dose inhaler). In some embodiments, the composition is adapted for ocular delivery, and the composition may further comprise a device for conjunctival administration (e.g., dropper or squeeze bottle). In some embodiments, the composition is adapted for intranasal administration, and the composition may further comprise a device for intranasal administration (e.g., dropper, pump spray, squeeze bottle, preservative-free airless spray or nasal pump device, e.g., nasal pump device with a reservoir bottle attached to an aerosolizer).

[0066] Intranasal drug delivery has been a topic of research and development for many years, but it is only within the last decade that carrier systems have been devised to effectively deliver substances (Sayani and Chien, Critical Reviews in Therapeutic Drug Carrier Systems 1996,13:85-184). Intranasal delivery has several beneficial features, including relatively high bioavailability, rapid absorption kinetics, and avoidance of first-pass effect in the liver. In some embodiments, intranasal administration can allow delivery of oxytocin peptide to the nasal cavity, and in other embodiments, intranasal administration can allow targeted delivery to the cranial nerves of the nose and / or brain. Without wishing to be bound by any particular theory, intranasal administration of oxytocin peptide can target either or both of the olfactory nervous system or the trigeminal nervous system. Oxytocin peptide can be delivered intranasally in any applicable form, including, but not limited to, liquid formulation, solid formulation (e.g., dry powder formulation), gel formulation, or emulsion formulation.

[0067] In embodiments in which the combination of oxytocin and magnesium ions is administered intranasally, the composition may be prepared as a liquid aerosol formulation combined with a dispersing agent and / or a physiologically acceptable diluent. Alternatively, dry powder aerosol formulations are contemplated, which may include the subject compound in finely divided solid form, and a dispersing agent that allows for rapid dispersion of the dry powder particles. In either liquid or dry powder aerosol formulations, the formulation is aerosolized into small liquid or solid particles to ensure that the aerosolized dose reaches the mucosa of the nasal passages or the lungs. The term "aerosol particles" is used herein to describe suitable liquid or solid particles of sufficiently small particle diameter for distribution in the nose (in the range of about 10 microns) or lungs (in the range of about 2-5 microns) to the targeted mucosa or alveolar membrane. Other considerations include the structure of the delivery device, additional components in the formulation, and the properties of the particles. These aspects of nasal or pulmonary administration of drugs are well known in the art, and the manipulation of the formulation, the means of aerosolization, and the structure of the delivery device are within the ordinary skill level of the art.

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

[0069] In some embodiments, the device for intranasal delivery is a nasal pump device. In some embodiments, the nasal pump device includes a reservoir bottle attached to a pump actuator. In some embodiments, the pump actuator meters to deliver 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) with a specific droplet size distribution. In some embodiments, the nasal pump device includes a reservoir bottle attached to an aerosolizer, such as an Equadel pump sold by Aptar Pharma. In some embodiments, the device for intranasal administration functions regardless of the pressure applied to the pump once a threshold is reached. In some embodiments, the device for intranasal administration is a mucosal spray device (e.g., LMA® MAD NASAL) that can be attached to a syringe. TMFor administration in larger mammals, the nasal pump device may include a reservoir bottle attached to a pump actuator that metering delivers a larger volume (e.g., from about 100 μL to about 600 μL or more).

[0070] In some embodiments, the device for intranasal delivery is designed to deliver multiple doses of a drug formulation. For example, the nasal pump device may include a reservoir bottle attached to a pump actuator, where the reservoir bottle holds multiple doses of a liquid formulation, and the pump actuator meters to deliver a specific volume that is a portion of the liquid formulation held in the reservoir bottle. In some embodiments, the pump actuator meters to deliver about 50 μL of the liquid formulation per spray. The nasal pump device may include a filter to prevent backflow to reduce the ingress of contaminants (e.g., bacteria) into the reservoir bottle. In some embodiments, the nasal pump device includes a metal-free flow path (e.g., a plastic flow path) for delivering the liquid formulation. In some embodiments, the pump device uses a plastic material that is stable to gamma radiation (used to sterilize the nasal device). In some embodiments, the device for intranasal delivery includes a multi-dose pump with a microbial filter and an automatic blocking mechanism on the pump actuator, such as the spray device described in U.S. Pat. No. 5,988,449.

[0071] In some embodiments, the device for intranasal delivery is a breath-actuated nasal delivery device (e.g., the device described in U.S. Patent Nos. 7,784,460 and 7,854,227). Such devices can improve delivery to target sites deep within the nasal cavity. In some embodiments, a standard metered dose spray device is incorporated into a housing where the patient breathes into the mouthpiece to activate the device. In some embodiments, the device comprises a cone-shaped sealed nosepiece and mouthpiece incorporating a conventional mechanical spray pump (e.g., the Equadel pump sold by Aptar Pharma), a chargeable spring, and a breath-activated mechanism. The system can be used for single or multiple dose delivery. One example of such a liquid delivery device is the OptiMist™ device sold by OptiNose. In use, the nosepiece of the device is inserted into the nostril and the patient breathes into the mouthpiece. This closes the soft palate, putting pressure on the nostrils and opening a passageway behind the nasal septum that allows air to flow, allowing air to exit through the other nostril (bi-directional flow). The device is activated by exhalation, so small particles cannot enter the lungs. Modifying the flow rate and particle size allows for targeting of specific nasal areas.

[0072] In some embodiments, the device for intranasal delivery is a unit dose metered spray device suitable for single administration of the magnesium-containing oxytocin peptide formulation or composition. In some embodiments, the device for intranasal delivery is a multi-dose metered spray pump device suitable for repeated administration of the oxytocin peptide.

[0073] Droplet size, plume volume and flow rate can be modified to target specific nasal regions. Liquid sprays can provide droplet sizes of 5-50 microns to target the olfactory epithelium and / or airway epithelium. Larger droplets fall primarily into the nasopharynx and are swallowed, while smaller droplets target lung tissue. Mass Median Equivalent Aerodynamic Diameter (MMAD) is used to specify droplet size. The pH of the nasal spray is optimized to deliver the charged peptide in a nearly ionized state. The nose typically tolerates solutions with a pH of about 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. Thus, the plume volume can be up to (and including) 100 μL. For use in large mammals, the plume volume can be up to (and including) 150 μL or greater (e.g., 600 μL or greater) in volume. For use in infants and children, or for veterinary use in small animals (e.g., rodents, cats), smaller plume volumes (5-50 μL) can be used.

[0074] In some embodiments, the device for intranasal delivery is ergonomically designed to facilitate patient compliance (e.g., a pump device with a side actuated trigger mechanism). In some embodiments, the device for intranasal delivery includes a metered-dose spray pump that operates as a closed system that does not allow air to enter the pump device and thus prevents contamination with airborne microorganisms. In some embodiments, the device for intranasal delivery includes a metered-dose spray pump that operates with a filter. Passing air is drawn through a filter assembled in the pump, thereby keeping airborne microorganisms out of the pump device. In some embodiments, the intranasal delivery device including the nasal pump device may further include a microelectronic device that may facilitate data transmission and monitoring of treatment.

[0075] In some embodiments, a magnesium-containing oxytocin peptide formulation or composition comprises an oxytocin peptide and magnesium ions, wherein the oxytocin peptide and magnesium ions are included in any one of the devices for intranasal delivery described herein, and the concentrations of the oxytocin peptide and magnesium ions are within any of the concentration ranges described herein, as if each and every combination of device and concentration were 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 non-verbal communication, and repetitive behaviors. With the publication of the Diagnostic and Statistical Manual of Mental Disorders, Fifth Edition (DSM-5) in May 2013, all autism disorders were combined into one umbrella 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 syndrome. See http: / / www.autismspeaks.org / what-autism. Those skilled in the art will recognize that symptoms of autism spectrum disorder overlap significantly with many other mental disorders. Examples of disorders that exhibit symptoms similar to those exhibited by autism spectrum disorders 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] DSM-5 provides diagnostic criteria for ASD that include: (A) persistent deficits in social communication and social interaction across multiple contexts, as manifested currently or historically by: (1) deficits in social-emotional reciprocity (ranging, e.g., from abnormal social approaches and failure to engage in normal conversational exchanges; to reduced sharing of interests, emotions, or attachments; to failure to initiate or respond to social interactions); (2) deficits in nonverbal communicative behaviors used in social interaction (e.g., from poor integration of verbal and nonverbal communication; to poor eye contact and body language). (B) Restricted, repetitive patterns of behavior, interests, or activities, as manifested by at least two of the following current or historical examples: (1) Stereotyped or repetitive movements, object use, or speech (e.g., simple motor stereotypies, etc.); and (3) Deficits in developing, maintaining, and understanding relationships (e.g., difficulty adjusting behavior to suit various social contexts, difficulty sharing creative play or making friends, etc., to lack of interest in peers). (2) persistence of sameness, rigid adherence to routines, or ritualized patterns or verbal and nonverbal behaviors (e.g., extreme distress at small changes, difficulties with transitions, rigid thought patterns, greeting rituals, need to take the same route or eat the same food every day); (3) highly localized and fixed interests that are abnormal in intensity or focus of interest (e.g., strong attachment to or preoccupation with unusual objects, overly localized or fixed interest);and (4) hyper- or hypo-sensitivity to sensory input or unusual interest in sensory aspects of the environment (e.g., seeming indifference to pain / temperature, adverse reactions to certain sounds or textures, excessive smelling or touching of objects, preoccupation with lights or movement). See http: / / www.autismspeaks.org / what-autism / diagnosis / dsm-5-diagnostic-criteria;

[0078] Autism spectrum disorder (ASD) is characterized by social interaction difficulties, 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 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, including anxiety and social and communication deficits in autism spectrum disorder. However, the effectiveness of oxytocin in treating social and communication deficits in autism spectrum disorder has been observed to vary widely between patients. It is believed that the variation in receptor availability and receptor affinity for oxytocin is the cause of the variation in effectiveness. Clinical efforts in treating ASD using commercially available formulations of oxytocin (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 for efficacy. The present invention provides methods for administering oxytocin peptides in more potent formulations and lower volumes such that an effective amount of the formulation can be delivered using a nasal device to treat autism spectrum disorder, a disorder manifesting one or more symptoms associated with autism spectrum disorder, or social and communication deficits.

[0080] In one aspect, a method for treating autism spectrum disorder, a disorder manifesting one or more symptoms associated with autism spectrum disorder, social and communication deficits, or anxiety is provided, comprising administering to a subject in need thereof an effective amount of oxytocin peptide and magnesium ion, wherein the effective amount is delivered via intranasal administration in a volume that is easily absorbed in the nasal cavity. In some embodiments, the volume that allows for easy absorption of oxytocin peptide and magnesium ion 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 ion 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 aspect, a method is provided that includes administering to a subject in need thereof an effective amount of oxytocin peptide and magnesium ions, 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 one or more symptomatic disorders 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. Co-administration of oxytocin and magnesium ions according to the present invention produces synergistic or enhanced improvements in social behavior and reduced anxiety relative to administration of oxytocin alone. The mechanisms underlying these effects are unclear, but appear to include either non-competitive blockade of N-methyl D-aspartate (NMDA) neurotransmitter molecule receptors, or increased affinity of oxytocin receptor action as an allosteric modulator, or both.

[0083] In some embodiments, a method is provided for treating an autism spectrum disorder, comprising administering to a subject in need thereof an effective dose of an oxytocin peptide and a magnesium ion, wherein the co-administration of the oxytocin peptide and the magnesium ion results in a synergistic or enhanced effect. In some embodiments, a method is provided for alleviating or reducing one or more symptoms associated with an autism spectrum disorder, comprising administering to a subject in need thereof an effective dose of an oxytocin peptide and a magnesium ion, wherein the co-administration of the oxytocin peptide and the magnesium ion results in a synergistic or enhanced effect. In some embodiments, a method is provided for treating a disorder manifesting one or more symptoms associated with an autism spectrum disorder, comprising administering to a subject in need thereof an effective dose of an oxytocin peptide and a magnesium ion, wherein the co-administration of the oxytocin peptide and the magnesium ion results in a synergistic or enhanced effect. In some embodiments, oxytocin peptide and magnesium ion are administered at a dose that produces a total effect on alleviating or reducing its symptoms that is greater than the sum of the effects of the equivalent doses of oxytocin peptide and magnesium salt administered separately.In some embodiments, oxytocin peptide and magnesium ion are administered at a dose that produces a faster onset of effect and / or a longer lasting effect than occurs after administration of each agent used alone in an equivalent amount.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 repetitive behaviors, limited interests and activities.Other behaviors and characteristics observed in individuals with autism spectrum disorder also include aversion to physical contact, generalized anxiety, monotony of vocalization or inability to regulate voice 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 exhibited in autism spectrum disorders 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 the loss of function of a gene located 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 emotional outbursts, stubbornness, manipulative behavior, and compulsive behaviors, including scratching the skin. Other symptoms often observed in individuals with Prader-Willi syndrome are persistent deficits in social communication and interaction, anxiety and irritability, and sleep problems.

[0085] In some aspects, a method for treating Prader-Willi syndrome is provided, comprising administering to a subject in need thereof an effective dose of oxytocin peptide and magnesium ion. In some embodiments, co-administration of the oxytocin peptide and the magnesium ion results in a synergistic or enhanced effect. In some embodiments, the oxytocin peptide and magnesium ion are administered in doses that result in a greater overall effect than the sum of the effects of equal doses of oxytocin peptide and magnesium salt administered individually. In some embodiments, the oxytocin peptide and magnesium ion are administered in doses that result in a faster onset of effect and / or a longer lasting effect than occurs following administration of the individual agents used alone in equal amounts. In one embodiment, the present invention provides a method for treating Prader-Willi syndrome, comprising administering to a subject in need thereof an effective dose of oxytocin peptide and magnesium ion, wherein the effective dose of oxytocin peptide and magnesium ion are administered intranasally in a liquid formulation, and the volume of the administered liquid formulation 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).

[0086] In some embodiments, methods are provided for alleviating or reducing one or more symptoms associated with Prader-Willi syndrome, comprising administering to a subject in need thereof an effective dose of oxytocin peptide and magnesium ion. 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, co-administration of the oxytocin peptide and the magnesium ion results in a synergistic or enhanced effect. In some embodiments, the oxytocin peptide and magnesium ion are administered in a dose that results in an overall effect on alleviating or reducing symptoms that is greater than the sum of the effects of equal doses of oxytocin peptide and magnesium salt administered individually. In some embodiments, the oxytocin peptide and magnesium ion are administered in a dose that results in a faster onset of effect and / or a longer lasting effect than occurs following administration of the individual agents 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, comprising administering to a subject in need thereof an effective dose of oxytocin peptide and magnesium ion, wherein the effective dose of oxytocin peptide and magnesium ion is administered intranasally in a liquid formulation, and the volume of the administered liquid formulation 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 ion 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 aspects, methods are provided for treating anxiety associated with Prader-Willi syndrome, comprising administering to a subject in need thereof an effective dose of an oxytocin peptide and magnesium ions. In some embodiments, co-administration of the oxytocin peptide and the magnesium ions results in a synergistic or enhanced effect. In some embodiments, the oxytocin peptide and magnesium ions are administered in doses that result in an overall effect on alleviating or reducing anxiety that is greater than the sum of the effects of equal doses of oxytocin peptide and magnesium salt administered individually. In some embodiments, the oxytocin peptide and magnesium ions are administered in doses that result in a faster onset of effect and / or a longer lasting effect than occurs following administration of the individual agents used alone in equal amounts. In one embodiment, the present invention provides a method for treating anxiety associated with Prader-Willi syndrome, comprising administering to a subject in need thereof an effective dose of oxytocin peptide and magnesium ion, wherein the effective dose of oxytocin peptide and magnesium ion are administered intranasally in a liquid formulation, and the volume of the administered liquid formulation 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 ion 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 social and communication deficits, comprising administering to a subject in need thereof an effective dose of oxytocin peptide and magnesium ion, wherein the simultaneous administration of the oxytocin peptide and the magnesium ion results in a synergistic or enhanced effect. In one embodiment, the present invention provides a method for treating anxiety, comprising administering to a subject in need thereof an effective dose of oxytocin peptide and magnesium ion, wherein the simultaneous administration of the oxytocin peptide and the magnesium ion results in a synergistic or enhanced effect. In some embodiments, the oxytocin peptide and magnesium ion are administered in a dose that results in an overall effect on reducing social and communication deficits and / or anxiety that is greater than the sum of the effects of equal doses of oxytocin peptide and magnesium salt administered individually. 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 social and communication deficit is an impairment in communication skills and / or social interaction, lack of eye contact, and / or an inability to form and / or maintain social relationships.

[0089] In some aspects, a method for treating anxiety associated with autism spectrum disorder is provided, comprising administering to a subject in need thereof an effective dose of oxytocin peptide and magnesium ion. In some embodiments, the simultaneous administration of the oxytocin peptide and the magnesium ion results in a synergistic or enhanced effect. In some embodiments, the oxytocin peptide and magnesium ion are administered in a dose that results in an overall effect on alleviating or reducing anxiety that is greater than the sum of the effects of equal doses of oxytocin peptide and magnesium salt administered individually. In some embodiments, the oxytocin peptide and magnesium ion are administered in a dose that results in a faster onset of effect and / or a longer lasting effect than occurs following administration of the individual agents used alone in equal amounts. In some embodiments, the molar ratio of the amount of oxytocin peptide to the amount of magnesium or magnesium ion 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] The oxytocin peptide and magnesium ions may be administered together or sequentially. In some embodiments, the oxytocin peptide is administered simultaneously with the magnesium ions in the same unit dose. In some embodiments, the oxytocin peptide is administered simultaneously with the magnesium ions but in separate unit doses or formulations. In some embodiments, the oxytocin peptide and magnesium ions are administered sequentially. In some embodiments, the magnesium ions are 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 10 minutes to about 2 hours after administration of the magnesium ions. In some of these embodiments, the oxytocin peptide is administered about 10 minutes to about 2 hours, about 10 minutes to about 1 hour, about 10 minutes to about 30 minutes, about 20 minutes to about 2 hours, about 20 minutes to about 1 hour, about 30 minutes to about 2 hours, or about 30 minutes to about 1 hour after administration of the magnesium ions. In some of these embodiments, the oxytocin peptide is administered about 10 minutes, about 15 minutes, about 20 minutes, about 30 minutes, about 45 minutes, about 60 minutes, about 90 minutes, or about 120 minutes after administration of the magnesium ions. In some of these embodiments, the oxytocin peptide is administered about 10 minutes, about 15 minutes, about 20 minutes, or about 30 minutes after administration of the magnesium ions. In one embodiment, the oxytocin peptide is administered to the subject first, and then the magnesium ions are administered to the subject. In some embodiments, the subject is a human.

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

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

[0093] In some embodiments, subjects are selected for treatment based on subjects having high levels of IL-6. The level of IL-6 may be high compared to a control or reference. In some embodiments, the level of IL-6 is high compared to a control or reference if it is significantly greater than the control or reference as determined by appropriate statistical analysis. In some embodiments, the level of IL-6 is high compared to a control or reference if it is at least one standard deviation greater than the control or reference. In some embodiments, the control is a value for the level of IL-6 as determined in age-matched and gender-matched healthy subjects. In some embodiments, the reference is a reported value for the level of IL-6 (e.g., a reported value for IL-6 in age-matched and gender-matched healthy subjects). In some embodiments, the level of IL-6 is determined as the level of IL-6 in a sample (such as a tissue or liquid sample) from the subject (including, but not limited to, whole blood, serum, plasma, tears, etc.). The level of IL-6 in the sample can be determined by any method known in the art (e.g., by immunoassay, e.g., ELISA-based assay). See, e.g., 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, a method for treating an autism spectrum disorder, a disorder manifesting one or more symptoms associated with an autism spectrum disorder, social and communication deficits, or anxiety includes measuring a level of IL-6 in a subject (e.g., a serum level of IL-6), and administering to a subject with high IL-6 levels an effective dose of an oxytocin peptide and magnesium ions.

[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 administering to a subject in need thereof an effective dose of oxytocin peptide and magnesium ion, wherein the simultaneous administration of the oxytocin peptide and the magnesium ion produces a synergistic or enhanced effect, further comprising administering to the subject an effective amount of interleukin-6 (IL-6). In some embodiments, the effective amount of IL-6 causes an increase in the expression of oxytocin receptor (OTR) in the subject.

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

[0097] The oxytocin peptide and magnesium ions may be administered to a subject in need thereof via the same or different routes. In some embodiments, the oxytocin peptide is administered via craniofacial mucosal administration (e.g., nasal, buccal, sublingual, or ocular administration). In one embodiment, both the oxytocin peptide and magnesium ions are administered intranasally in the same formulation. In one embodiment, the oxytocin peptide is administered via craniofacial mucosal administration and the magnesium ions are administered systemically, e.g., 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 ion are administered via intranasal administration. The oxytocin peptide and / or magnesium ion can be administered to the mucosal tissue in the nasal cavity using a suitable device for intranasal delivery (e.g., a nasal delivery device described herein). Suitable areas in 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 ion is administered to the upper one-third of the nasal cavity. In some embodiments, the oxytocin peptide and / or magnesium ion is administered to the lower two-thirds of the nasal cavity. In some embodiments, the oxytocin peptide and / or magnesium ion is specifically administered to reach both the lower two-thirds and the upper one-third of the nasal cavity. In some embodiments, there is provided a method for treating autism spectrum disorder, one or more symptoms associated with autism spectrum disorder, or a disorder that manifests one or more symptoms associated with autism spectrum disorder, comprising administering an effective dose of oxytocin peptide and magnesium ion to a subject in need thereof intranasally, wherein the simultaneous administration of the oxytocin peptide and the magnesium ion produces synergistic or enhanced effect.In some embodiments, the method is for treating social and communication deficits, or anxiety.

[0099] In some embodiments, according to any of the methods described herein in which IL-6 is administered to a subject, IL-6 is administered via intranasal administration. IL-6 can be administered to mucosal tissue in 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, an effective dose of oxytocin peptide is about 4 μg to about 1000 μg, about 8 μg to about 1000 μg, about 8 μg to about 800 μg, about 8 μg to about 500 μg, about 8 μg to about 400 μg, about 8 μg to about 300 μg, about 8 μg to about 200 μg, about 8 μg to about 100 μg, about 8 μg to about 80 μg, about 8 μg to about 50 μg, about 10 μg to about 1000 μg, about 10 μg to about 500 μg, about 10 μg to about 200 μg, about 10 μg to about 100 μg, about 16 μg to about 1000 μg, about 16 μg to about 800 μg, about 16 μg to about 500 μg, about 16 μg to about 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 1 000 μg, about 30 μg to about 500 μg, about 30 μg to about 300 μg, about 30 μg to about 120 μg, about 30 μg to about 90 μg, about 50 μg to about 1000 μg, about 50 μg to about 500 μg, about 50 μg to about 250 μg, about 50 μg to about 100 μg, or about 50 μg to about 80 μg. In some embodiments, the effective dose of the 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 a preferred embodiment, the effective dose of the 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, an effective dose of the oxytocin peptide is about 0.25 IU to about 1000 IU. In some embodiments, an effective dose of the 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, an effective dose of oxytocin peptide is from about 2 IU to about 500 IU, from about 4 IU to about 500 IU, from about 4 IU to about 400 IU, from about 4 IU to about 250 IU, from about 4 IU to about 200 IU, from about 4 IU to about 150 IU, from about 4 IU to about 100 IU, from about 4 IU to about 50 IU, from about 4 IU to about 40 IU, from about 4 IU to about 25 IU, from about 5 IU to about 500 IU, from about 5 IU to about 250 IU, from about 5 IU to about 100 IU, from about 5 IU to about 50 IU, from about 8 IU to about 500 IU, from about 8 IU to about 400 IU, from about 8 IU to about 250 IU, from about 8 IU to about 200 IU , about 8 IU to about 100 IU, about 8 IU to about 80 IU, about 8 IU to about 60 IU, about 8 IU to about 40 IU, about 10 IU to about 500 IU, about 10 IU to about 400 IU, about 10 IU to about 250 IU, about 10 IU to about 100 IU, about 10 IU to about 50 IU, about 15 IU to about 500 IU, about 15 IU to about 250 IU, about 15 IU to about 150 IU, about 15 IU to about 60 IU, about 15 IU to about 45 IU, about 25 IU to about 500 IU, about 25 IU to about 250 IU, about 25 IU to about 125 IU, about 25 IU to about 50 IU, or about 25 IU to about 40 IU. In some embodiments, the effective dose of the 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 the 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 the combination is effective in one embodiment to provide a clinically measurable improvement of symptoms of autism spectrum disorder or related disorder. The combination of oxytocin and magnesium ion provides a synergistic or enhanced effect of improving autism spectrum disorder or related disorder. In some embodiments, oxytocin is administered at a sub-therapeutic effective dose relative to the dose of oxytocin administered as a single agent. The dose of oxytocin as a single agent depends in part on the route of administration. Thus, the dose of oxytocin in the combination therapy described herein also depends in part on the route of administration.

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

[0104] In some embodiments, the magnesium salt administered comprises magnesium chloride and an effective dose of magnesium salt is about 0.48 mg to about 600 mg of magnesium chloride hexahydrate (MgCl2·6H2O, MW 203.3). In some embodiments, an effective dose of magnesium chloride hexahydrate is about 0.48 mg to about 300 mg, about 0.5 mg to about 150 mg, about 0.5 mg to about 75 mg, about 5 mg to about 150 mg, about 5 mg to about 75 mg, about 5 mg to about 50 mg, about 10 mg to about 600 mg, about 10 mg to about 300 mg, about 10 mg to about 150 mg, about 10 mg to about 75 mg, about 10 mg to about 50 mg, about 10 mg to about 30 mg, or about 12 mg to about 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 about 0.48 mg to about 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 about 0.48 mg to about 300 mg, about 0.5 mg to about 150 mg, about 0.5 mg to about 75 mg, about 5 mg to about 150 mg, about 5 mg to about 75 mg, about 5 mg to about 50 mg, about 10 mg to about 600 mg, about 10 mg to about 300 mg, about 10 mg to about 150 mg, about 10 mg to about 75 mg, about 10 mg to about 50 mg, about 10 mg to about 30 mg, or about 12 mg to about 24 mg. In some preferred embodiments, the effective dose of magnesium citrate (eg, 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, an effective dose of magnesium citrate is about 0.48 mg to about 12 mg, about 0.5 mg to about 10 mg, about 0.5 mg to about 8 mg, about 0.5 mg to about 5 mg, about 0.5 mg to about 2.5 mg, about 0.5 mg to about 1 mg, about 1 mg to about 10 mg, about 1 mg to about 8 mg, about 1 mg to about 5 mg, about 1 mg to about 2 mg, about 2 mg to about 10 mg, about 2 mg to about 8 mg, about 2 mg to about 6 mg, about 2 mg to about 4 mg, about 3 mg to about 10 mg, about 4 mg to about 10 mg, about 4 mg to about 8 mg, about 4 mg to about 6 mg, about 5 mg to about 10 mg, about 5 mg to about 8 mg, about 5 mg to about 7 mg, about 5 mg to about 6 mg, about 6 mg to about 10 mg, about 6 mg to about 8 mg, or about 6 mg to about 7 mg. When other magnesium salts are used in place of magnesium citrate, an effective dose of the magnesium salt provides an amount of magnesium ions equivalent to that provided by magnesium citrate.

[0106] It is intended and understood that each and every dosage of magnesium ion described herein may be combined with each and every dosage of oxytocin peptide described herein as if each and every combination were set forth individually. For example, in some embodiments, the effective dosage of oxytocin peptide is about 0.5 μg to about 2000 μg, and the effective dosage of magnesium ion is about 50 μg to about 68 mg of magnesium. In some embodiments, the effective dosage of oxytocin peptide is about 15 μg to about 120 μg (e.g., about 60 μg or about 66 μg), and the effective dosage of magnesium ion is equal to the amount of magnesium ion 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, there is provided a method for treating an autism spectrum disorder, a disorder manifesting one or more symptoms associated with an autism spectrum disorder, social and communication deficits, or anxiety, comprising administering to a subject in need thereof (e.g., by intranasal administration) an effective dose of an oxytocin peptide and magnesium ions, 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 for magnesium-containing oxytocin peptide formulations or compositions. In some embodiments, a method is provided for treating an autism spectrum disorder, a disorder manifesting one or more symptoms associated with an autism spectrum disorder, social and communication deficits, or anxiety, comprising administering to a subject in need thereof an effective dose of an oxytocin peptide and magnesium ions (e.g., by intranasal administration), wherein the molar ratio of the dose of oxytocin peptide administered to the dose of magnesium ions administered is , about 1:40 to about 1:40000, preferably about 1:80 to about 1:8000, more preferably about 1:175, about 1:280, about 1:500, about 1:560, about 1:800, about 1:1000, about 1:1100, about 1:1200, about 1:1400, about 1:1600, about 1:1700, about 1:1800, about 1:2000, about 1:2400, about 1:3000, or any of the OT / Mg(m) ratios described herein for 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 the embodiments, the magnesium ions are provided by magnesium citrate and / or magnesium chloride.In some of these embodiments, the social and communication deficit is an impairment in communication skills and / or social interaction, lack of eye contact, and / or an inability to form and / or maintain social relationships.

[0108] In one embodiment, a method is provided for treating an autism spectrum disorder, a disorder manifesting one or more symptoms associated with an autism spectrum disorder, social and communication deficits, or anxiety, comprising administering intranasally to a subject in need thereof an oxytocin peptide at 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 at 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 intranasally administering to a subject in need thereof an effective amount of a magnesium-containing oxytocin peptide formulation or composition described herein. In one embodiment, the method includes administering to a subject in need thereof an effective amount of an oxytocin peptide and magnesium ions intranasally in a liquid formulation having a volume of about 5 μL to about 1000 μL. In one embodiment, the method includes administering to a subject in need thereof an effective dose of a magnesium-containing oxytocin peptide formulation or composition comprising 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. In one embodiment, the method includes administering into the nasal cavity of a subject in need thereof an effective dose of a magnesium-containing oxytocin peptide formulation comprising 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.In one embodiment, the method includes administering into the nasal cavity of a subject in need thereof an effective dose of a magnesium-containing oxytocin peptide formulation comprising 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. In one embodiment, the method includes administering into the nasal cavity of a subject in need thereof an effective dose of a magnesium-containing oxytocin peptide formulation comprising 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. In one embodiment, the method includes administering into the nasal cavity of a subject in need thereof an effective dose of a magnesium-containing oxytocin peptide formulation comprising 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.

[0109] In some embodiments, an effective dose of oxytocin peptide and magnesium ions comprises 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 comprises 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 are administered intranasally in a liquid formulation, and the volume of the liquid formulation administered is about 5 μL to about 1000 μL. In some embodiments, the volume administered is about 5 μL to about 500 μL, about 5 μL to about 250 μL, about 5 μL to about 100 μL, about 5 μL to about 50 μL, about 10 μL to about 1000 μL, about 10 μL to about 500 μL, about 10 μL to about 250 μL, about 10 μL to about 100 μL, about 25 μL to about 1000 μL, about 25 μL to about 500 μL, about 25 μL to about 250 μL, about 25 μL to about 100 μL, about 50 μL to about 1 000 μL, about 50 μL to about 750 μL, about 50 μL to about 500 μL, about 50 μL to about 450 μL, about 50 μL to about 400 μL, about 50 μL to about 350 μL, about 50 μL to about 300 μL, about 50 μL to about 250 μL, about 50 μL to about 200 μL, about 50 μL to about 150 μL, about 100 μL to about 500 μL, about 100 μL to about 400 μL, about 100 μL to about 300 μL, or about 100 μL to about 200 μL. In some embodiments, the volume administered 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 combination of oxytocin peptides and magnesium ions described herein can be used for the treatment of any social and communication deficit treatable by oxytocin (e.g., impairment in communication skills and / or social interaction, lack of eye contact, and / or inability to form and / or maintain social relationships).Therefore, a method for treating a social and communication deficit is provided, comprising administering to a subject in need thereof an effective dose of oxytocin peptides and magnesium ions, wherein the social and communication deficit is impairment 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 administering to a subject in need thereof an effective dose of oxytocin peptides and magnesium ions intranasally. 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 manifesting one or more symptoms associated with autism spectrum disorder, social and communication deficits, or anxiety, comprising administering an effective dose of an oxytocin peptide and magnesium ions intranasally to a subject in need thereof (e.g., a human or veterinary patient), wherein co-administration of the oxytocin peptide and the 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, an 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), more preferably about 15 μg (or 7.5 IU) to about 120 μg (or 60 IU). In some embodiments, an effective dose of magnesium ion is about 50 μg to about 68 mg. In some embodiments, magnesium ion is provided using a magnesium salt (e.g., magnesium chloride and / or magnesium citrate) administered in an amount providing about 50 μg to about 68 mg of magnesium. In some embodiments, an effective dose of magnesium ion is provided by using about 0.48 mg to about 600 mg of magnesium citrate. In some embodiments, an effective dose of magnesium ion 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, the effective dose of oxytocin peptide and magnesium ions comprises about 10 μg to about 120 μg (e.g., about 66 μg) oxytocin peptide administered in an aqueous solution comprising about 10% to about 14% (e.g., about 12%) (w / v) magnesium citrate.

[0113] kit A kit is provided herein for carrying out any of the methods described herein. The kit is provided for use in treating autism spectrum disorder, one or more symptoms associated with autism spectrum disorder, social and communication deficits, or anxiety. In some embodiments, the kit includes oxytocin peptide and magnesium ion in suitable packaging, wherein the oxytocin peptide and the magnesium ion are in an amount that produces a synergistic or enhanced effect when used in treating autism spectrum disorder, one or more symptoms associated with autism spectrum disorder, social and communication deficits, or anxiety, and a device for craniofacial mucosal administration (e.g., intranasal administration). The kit may further include a protease inhibitor and / or at least one absorption enhancer. The kit may further include IL-6. Other kits may further include instructions that provide information to a user and / or a health care provider to carry out any of the methods described herein. The kit may further comprise reagents / tools for measuring IL-6 levels in a subject; and, optionally, instructions for predicting the efficacy of intranasal oxytocin and magnesium ions.

[0114] Also provided are kits that include the magnesium-containing oxytocin peptide formulations described herein contained in a device for craniofacial mucosal administration (e.g., a device for intranasal administration, such as a nasal pump device) and suitable packaging material. The kits may further include instructions for administering the magnesium-containing oxytocin peptide formulation to a subject in need thereof.

[0115] Instructions for use of a kit to practice the invention will typically describe how to use the contents of the kit to practice the method of the invention. Instructions provided with a kit of the invention will typically be written instructions on a label or insert (e.g., a sheet of paper included with the kit), although machine-readable instructions (e.g., instructions embodied on a magnetic or optical storage disk) are also acceptable. EXAMPLES

[0116] The present invention may be further understood by reference to the following examples, which are provided for purposes of illustration and are not intended to be limiting.

[0117] Example 1: Exemplary preparation of magnesium-containing oxytocin peptide formulations Example 1A The drug product formulation, which is hypertonic and targets a pH of 4.5, consists of oxytocin USP (150 IU / mL); magnesium chloride USP (as the hexahydrate or anhydrous salt); citric acid USP (as the anhydrous or monohydrate form); sodium hydroxide NF; and sterile water for injection USP. The quantitative composition is provided in Table 1. 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 monographs compendium (USP / NF). [Table 1]

[0118] The drug product is manufactured by dissolving the ingredients in sterile water for injection, sterile filtering, filling into snap-top vials in a preservative-free pump, and tested generally in accordance with the FDA's July 2002 nasal spray guidelines.

[0119] In one example, a 10 L batch of magnesium-containing oxytocin formulation according to the composition provided in Table 1 was prepared as follows: A formulation container was filled with water to approximately 60% of the required batch volume. With stirring at ambient temperature, the required amounts of sodium chloride, citric acid, and magnesium chloride hexahydrate were added in the following order: The materials dissolved easily. No heating was required, just gentle stirring. The pH of the solution was adjusted to 4.5 by adding 1N NaOH (in case of over-titration, one can back-titrate to pH 4.5 with 10% HCl). The required amount of oxytocin was added and stirred until dissolved. Water was added to bring the batch to final weight / volume. The solution was stirred until homogenous.

[0120] Example 1B The drug product formulation, which is isotonic and targets a pH of 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 monographs compendium (USP / NF).

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

[0122] For use as clinical material, oxytocin and excipients are manufactured according to current Good Manufacturing Practice and terminally sterilized (sterile filtration through a 0.2 micron membrane filter) before being filled into glass storage bottles and sealed with a pump actuator. By increasing or decreasing the amount of oxytocin, various formulation concentrations can be generated from this example. This 1 mL batch volume will yield approximately 10 doses of oxytocin. [Table 2]

[0123] Example 2: Rat model of social behavior Rats were treated intranasally with 20 μl (10 μl / nostril) of a solution containing saline, 10 μg oxytocin, a combination of 12% magnesium citrate and 10 μg oxytocin (approximately 1:1127 molar ratio of oxytocin to magnesium ion), or 12% magnesium citrate. Eight (8) rats were used in each treatment group. Forty minutes after intranasal drug administration, two animals from the same treatment group were paired and placed into the test chamber, and their social behaviors (sniffing, chasing, crawling up and down, social grooming [grooming the partner], and play fighting) were recorded for 10 minutes. The time spent in social interaction is shown in FIG. 1. The results show evidence of the enhanced effect of the combination of 12% magnesium citrate and 10 μg oxytocin on improving social behavior.

[0124] Example 3: Rat model of anxiety Example 3A Rats were treated intranasally with 20 μl (10 μl / nostril) of a solution containing saline, 10 μg oxytocin, 12% magnesium citrate and 10 μg oxytocin (approximately 1:1127 molar ratio of oxytocin to magnesium ion), or 12% magnesium citrate. Eight (8) 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 the open arms during a five minute period. The observed number of open arm entries is shown in FIG. 2. The results show evidence of a synergistic effect of the combination of 12% magnesium citrate and 10 μg oxytocin on reducing anxiety.

[0125] Example 3B Rats were treated intranasally with 20 μl (10 μl / nostril) of a solution containing saline, 3% magnesium citrate, 6% magnesium citrate, 16 μg oxytocin, 10 μg oxytocin, a combination of 3% magnesium citrate and 16 μg oxytocin (at a molar ratio of oxytocin to magnesium ion of approximately 1:176), or a combination of 6% magnesium citrate and 10 μg oxytocin (at a molar ratio of oxytocin to magnesium ion of approximately 1:563). Eight (8) rats were used in each treatment group. Thirty minutes after intranasal drug administration, the animals were exposed to elevated platform stress for 5 minutes and immediately placed into an elevated 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 they entered the open arms is shown in Figures 3A and 3B. Animals treated with a combination of 3% magnesium citrate and 16 μg oxytocin (approximately 1:176 molar ratio of oxytocin to magnesium ion) had less anxiety, as indicated by increased open arm entries, than animals treated with either 3% magnesium citrate alone or 16 μg oxytocin alone. In contrast, animals treated with 6% magnesium citrate and 10 μg oxytocin had less anxiety, as indicated by increased open arm entries. Animals treated with the oxytocin combination (oxytocin to magnesium ion in a molar ratio of approximately 1:563) showed a 6% reduction in open arm entries. had greater anxiety than animals treated with either magnesium citrate alone or 10 μg oxytocin alone.

[0126] To further assess anxiety, the latency to enter the open arms, the time spent in the open arms, and the number of entries into the closed arms are determined.

[0127] The experiment is repeated with additional amounts of magnesium citrate and oxytocin, including, for example, 6% magnesium citrate alone, 20 μg oxytocin alone, and a combination of 6% magnesium citrate and 20 μg oxytocin (approximately a 1:281 molar ratio of oxytocin to magnesium ion).

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

[0129] Example 5: Human Clinical Trials Using a placebo-controlled, randomized, double-blind, parallel design, the effects of a six-week course of twice-daily intranasal treatment with a combination of oxytocin and magnesium will be tested in male and female subjects aged 18-55 years diagnosed with autism spectrum disorder. 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) change in communication and restricted repetitive behavior scores as measured by the Autism Diagnostic Observation Test-II before and at the completion of the double-blind treatment period; (2) change in anxiety as measured by the State-Trait Anxiety Scale assessed before and at the completion of the double-blind treatment period; (3) change 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) change in gaze to the field atmosphere assessed before and at the completion of the double-blind treatment period; (5) changes in facial and vocal expressions analyzed from video recordings every 2 weeks during the study period; and (6) Changes in Clinical Global Impression and Global Rating of Functioning scales assessed every 2 weeks during the study period.

[0130] Study inclusion criteria consisted of the following: 1) diagnosed with autism spectrum disorder according to DSM-V; 2) above the cutoff for qualitative abnormality in reciprocal social interaction (Domain A) on the Autism Diagnostic Observation-Revised; and 3) a verbal IQ of greater than 85 and a full-scale IQ of greater than 80 as measured by the Wechsler Adult Intelligence Scale-III.

[0131] Study exclusion criteria consisted of the following: 1) major psychiatric diagnosis other than inclusion criterion 1); 2) current instability due to comorbid psychiatric diagnoses; 3) history of changes in psychotropic medication or drug dosage within 1 month of randomization; 4) currently being treated with more than two categories of psychotropic medication; 5) currently being treated with atomoxetine or methylphenidate; 6) history of ongoing oxytocin treatment; 7) history of sensitivity to oxytocin; 8) History of stroke or traumatic brain injury with loss of consciousness longer than 5 minutes; and 9) History of alcoholism or substance abuse or addiction.

[0132] Example 6: Effects of oxytocin and magnesium in social anxiety disorder Subjects meeting Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition criteria for generalized social phobia will be randomized to treatment with 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 1-week single-blind placebo run-in period, patients will receive an 11-week double-blind course of Treatments A, B, C, or D. Patients will receive two treatments daily, approximately 12 hours apart.

[0134] Optionally, serum IL-6 levels will be examined at the end of the 1-week run-in period and at the end of the 11-week course.

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

[0136] The effects of treatment groups are analyzed.

[0137] Exemplary embodiments The present invention is further illustrated by the following embodiments, the features of each of which may be combined, where appropriate and permissible, with any of the other embodiments.

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

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

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

[0141] Embodiment 4. In a further embodiment of any one of embodiments 1-3, 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, said oxytocin peptide and said magnesium ions are administered via intranasal administration.

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

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

[0146] Embodiment 9. In a further embodiment of any one of embodiments 1-8, 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 ion has an oxytocin:magnesium molar ratio of about 1:40 to about 1:40000.

[0149] Embodiment 12. In a further embodiment of any one of embodiments 1-11, the method is for treating an autism spectrum disorder.

[0150] Embodiment 13. In a further embodiment of any one of embodiments 1-11, the method is for treating a disorder that manifests one or more symptoms associated with an 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 a further embodiment of any one of embodiments 1 to 11, the method is for treating social and communication deficits.

[0153] Embodiment 16. In a further embodiment of any one of embodiments 1-11, the method is for treating anxiety.

[0154] Embodiment 17. In a further embodiment of any one 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 an autism spectrum disorder, a disorder manifesting one or more symptoms associated with an autism spectrum disorder, social and communication deficits, or anxiety, comprising administering to a subject in need thereof an effective dose of an oxytocin peptide and magnesium ions, wherein the effective dose of the oxytocin peptide and magnesium ions are administered intranasally in a liquid formulation, and the volume of the administered liquid formulation is from 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 from about 0.5 μg to about 2000 μg.

[0157] Embodiment 20. In a further embodiment of embodiment 18, the effective dose of magnesium ion is from about 50 μg to about 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 ion has an oxytocin:magnesium molar ratio of about 1:40 to about 1:40000.

[0160] Embodiment 23. In a further embodiment of embodiment 21 or 22, the volume of the liquid formulation administered is from about 50 μL to about 200 μL.

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

[0162] Embodiment 25. In a further embodiment of any one of embodiments 18-24, the method is for treating an autism spectrum disorder.

[0163] Embodiment 26. In a further embodiment of any one of embodiments 18-24, the method is for treating a disorder that manifests one or more symptoms associated with an 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 a further embodiment of any one of embodiments 18-24, the method is for treating social and communication deficits.

[0166] Embodiment 29. In a further embodiment of any one of embodiments 18-24, the method is for treating anxiety.

[0167] Embodiment 30. In a further embodiment of any one 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 apparatus.

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

[0171] Embodiment 34. In a further embodiment of embodiment 33, said pump actuator is metered to deliver a specific volume of about 50 μL.

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

[0173] Embodiment 36. In a further embodiment of any one of embodiments 32 to 35, the nasal pump device comprises: (i) a filter to prevent backflow; (ii) a metal-free flow path; and (iii) Gamma-ray-stable plastic materials It has one or more of the following:

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

[0175] Embodiment 38. In a further embodiment of embodiment 37, said 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 comprising from 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 comprising the magnesium in an amount providing from 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 from about 1:40 to about 1:800.

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

[0181] Embodiment 44. In a further embodiment of any one of embodiments 37 to 43, the device further comprises a device for administration to the craniofacial mucosa.

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

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

[0184] Although the foregoing invention has been described in some detail for purposes of illustration and example, for purposes of clarity of understanding, it will be apparent to those skilled in the art that certain changes and modifications can be made without departing from the invention. Therefore, such descriptions and illustrations should not be construed as limiting the scope of the invention.

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

Claims

1. 1. A combination for treating social anxiety disorder, social communication deficits, or social interaction deficits associated with autism spectrum disorder or Prader-Willi syndrome in a subject in need thereof, said combination being formulated as an aqueous solution suitable for craniofacial mucosal administration, said combination comprising an oxytocin peptide in an amount of 8 μg to 1000 μg, and a magnesium salt providing 0.11% to 1.65% (w / v) magnesium ion in said aqueous solution, wherein the molar ratio of oxytocin peptide to magnesium ion is about 1:1000 to about 1:1600.

2. The combination of claim 1 , wherein the craniofacial mucosal administration is intranasal administration.

3. 2. The combination of claim 1, wherein the effective dose of the oxytocin peptide is from about 8 μg to about 80 μg.

4. A combination according to any one of claims 1 to 3, wherein the magnesium salt is magnesium chloride hexahydrate.

5. A combination according to any one of claims 1 to 3, wherein the magnesium salt is magnesium citrate.

6. 6. The combination according to any one of claims 1 to 5, wherein the social anxiety disorder, deficit in social communication or deficit in social interaction is associated with Prader-Willi syndrome.

7. 7. The combination of claim 6, wherein the administration of the combination treats social anxiety disorder associated with Prader-Willi syndrome.

8. 7. The combination of claim 6, wherein the administration of the combination treats social communication deficits associated with Prader-Willi syndrome.

9. 7. The combination of claim 6, wherein the administration of the combination treats social interaction deficits associated with Prader-Willi syndrome.

10. The combination according to any one of claims 1 to 9, wherein the oxytocin peptide is human oxytocin (SEQ ID NO: 1).

11. The combination according to any one of claims 1 to 10, wherein the aqueous solution is administered in a device for intranasal administration.

12. The combination of claim 11, wherein the device is a nasal pump device.

13. 13. The combination of claim 12, wherein the nasal pump device is a metered volume nasal pump.

14. 13. The combination of claim 12, wherein the nasal pump device comprises a reservoir bottle attached to a pump actuator.

15. 15. The combination of claim 14, wherein the pump actuator is metered to deliver a specific volume between 50 μL and 200 μL.

16. 16. The combination of claim 15, wherein the pump actuator is metered to deliver a specific volume of 50 μL or 100 μL.

17. 17. The combination of claim 16, wherein the pump actuator meters to deliver the aqueous solution in 1 to 4 units of 50 μL / unit.

18. 18. The combination of claim 17, wherein the pump actuator meters to deliver the aqueous solution at 1-2 units of 100 μL / unit.

19. 20. The combination of claim 18, wherein the nasal pump device comprises a reservoir bottle attached to an aerosolizer.

20. The nasal pump device comprises: (i) a filter to prevent backflow; (ii) a metal-free flow path; and (iii) Gamma-ray stable plastic materials The combination according to any one of claims 11 to 19, comprising one or more of:

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