Labeled oxytocin and methods of making and using

Abstract

To provide a labeled oxytocin and a method for producing and using the same.SOLUTION: Disclosed are 13N-oxytocin molecules, methods of making 13N-oxytocin molecules, and methods of using 13N-oxytocin molecules in determining the distribution and kinetics following craniofacial or other administration of 13N-oxytocin molecules. The synthesis of radiolabeled oxytocin peptides remains a challenge and an impediment to using conventional imaging techniques such as PET. Provided herein are means to solve this problem by providing novel 13N-labeled oxytocin peptides and methods of making the same.SELECTED DRAWING: None

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of priority to U.S. Provisional Application No. 62 / 637,187, filed March 1, 2018, the contents of which are incorporated herein by reference in their entirety. Submitting a sequence listing as an ASCII text file

[0002] The contents of the following ASCII text file submission: Sequence Listing in Computer Readable Format (CRF) (Filename: 623632001640SEQLIST.TXT, Date Recorded: March 1, 2019, Size: 1 KB) are incorporated herein by reference in their entirety. Technical Field

[0003] The present invention provides 13 a composition comprising an N radiolabeled oxytocin peptide; 13 1. Methods for producing N-radiolabelled oxytocin peptides for use in positron emission tomography (PET) and other applications, including methods for producing N-radiolabelled oxytocin peptides 13 N radiolabeled oxytocin peptide. [Background technology]

[0004] Oxytocin is a naturally occurring nine-amino acid polypeptide implicated in several physiological bodily functions, including uterine contractions, muscle regeneration, and lactation. Additionally, oxytocin has been shown to play an important role in several peripheral and central nervous system functions. For example, oxytocin has been shown to be involved in pain, autism, appetite, anxiety, trust, blood sugar control, and interpersonal relationships.

[0005] Published articles have demonstrated that craniofacial mucosal, e.g., intranasal, administration of oxytocin can modulate these functions. For example, intranasal oxytocin administration has been shown to improve social function in autistic patients and relieve pain in migraine patients. However, no information is available describing the spatiotemporal distribution of oxytocin within the body after craniofacial mucosal application. Unlike measuring blood levels of oxytocin, measuring oxytocin levels in tissues, particularly the nervous system, after oxytocin administration is inherently difficult. Evaluating tissue levels of oxytocin after craniofacial or other application would allow for the determination of tissue oxytocin levels, which are crucial for therapeutic efficacy. Summary of the Invention [Problem to be solved by the invention]

[0006] One method for assessing the biodistribution of administered oxytocin is by positron emission tomography (PET). PET is a nuclear medicine imaging technique that produces images or photographs of functional processes in the body. This system detects gamma-ray pairs indirectly emitted by positron-emitting radionuclides (tracers, radioactive tracers, radiopharmaceuticals, etc.) that are introduced into the body on bioactive molecules. A radionuclide (or radioactive nuclide) is an atom with an unstable nucleus characterized by available excess energy that is donated to newly generated radioactive particles within the nucleus or via internal conversion. During this process, a radionuclide is said to undergo radioactive decay, resulting in the emission of gamma rays and / or subatomic particles such as alpha or beta particles. Such radiation constitutes ionizing radiation. Radionuclides are often referred to as radioactive isotopes (or radioisotopes). The three-dimensional distribution of radionuclide concentrations within the body can be constructed by computer analysis during the PET process. However, the synthesis of radiolabeled oxytocin peptides for use as radiotracers in PET remains challenging.

[0007] The disclosures of all publications, patents, patent applications, and published patent applications cited by reference herein are hereby incorporated by reference in their entirety. [Means for solving the problem]

[0008] Brief summary of the invention This application is 13 N-oxytocin, 13 a composition comprising N-oxytocin; 13 How to use N-oxytocin, and 13 A method for producing N-oxytocin is provided.

[0009] In some embodiments, the amino acid sequence of SEQ ID NO: 1 13N-labeled oxytocin peptides are provided. In some embodiments, one component of the oxytocin peptide is 14 The N atom is 13 In some embodiments, one residue of the oxytocin peptide is replaced with a N radionuclide. 13 In some embodiments, one residue of the oxytocin peptide is modified to include a moiety containing a N radionuclide. 13 In some embodiments, the amino acid is modified to include NH. 13 N-labeled oxytocin peptides are single 13 Contains N radionuclides.

[0010] The above 13 In some embodiments according to any of the N-labeled oxytocin peptides, 13 The N-labeled oxytocin peptides were modified at a) the glutamine residue at position 4 (SEQ ID NO: 2), b) the asparagine residue at position 5 (SEQ ID NO: 3), or c) the glycine residue at position 9 (SEQ ID NO: 4). 13 Contains N radionuclides.

[0011] In some embodiments, 13 The N-labeled oxytocin peptide is a compound of formula (IV).

[0012] In some embodiments, the amino acid sequence of SEQ ID NO: 1 13 1. A method for preparing an N-labeled oxytocin peptide, comprising: a) reacting a compound of formula (Ib) with diethyl cyanophosphonate (DECP) to provide a compound of formula (IIb); and b) treating the compound of formula (IIb) with a gas. 13 NH to provide a compound of formula (IIIb); and c) deprotecting the compound of formula (IIIb); 13 providing an N-labeled oxytocin peptide, 13A method is provided in which the N-labeled oxytocin peptide is a compound of formula (IV). In some embodiments, the reaction of the compound of formula (Ib) with DECP is carried out in the presence of dimethyl sulfoxide (DMSO), tetrahydrofuran (THF), and pentamethylpiperidine (PMP). In some embodiments, the amount of DECP is between about 0.7 equivalents and about 1.1 equivalents. In some embodiments, the ratio of DMSO to THF is between about 1:7 and about 1:11. In some embodiments, the deprotection comprises reacting the compound of formula (IIIb) with HCl / dioxane.

[0013] In some embodiments according to any of the methods described above, the method further comprises, in step c), 13 The method further comprises purifying the N-labeled oxytocin peptide to remove reagents, organic solvents, and precursors. In some embodiments, the purification comprises purification by solid phase extraction (SPE). In some embodiments, the purification by SPE comprises: a) 13 The N-labeled oxytocin peptide is applied to a first hydrophobic SPE column, 13 allowing N-oxytocin and precursors to be retained on a first hydrophobic SPE column; and b) applying a solution comprising an aqueous ion-pairing reagent to the first hydrophobic SPE column; 13 allowing N-oxytocin to elute in a first eluate; and c) applying the first eluate to a second hydrophobic SPE column to obtain a second hydrophobic SPE column. 13 allowing N-oxytocin to be retained on a second hydrophobic SPE column; and d) 13 and eluting N-oxytocin in a second eluent. In some embodiments, the first and / or second hydrophobic SPE columns comprise a strongly hydrophobic, silica-based bonded phase. In some embodiments, the solution comprising the aqueous ion-pairing reagent comprises between about 15% and about 25% acetonitrile.

[0014] In some embodiments, the composition is prepared by a process comprising a method according to any of the methods described above. 13 N-labeled oxytocin peptides are provided.

[0015] In some embodiments, there is provided a method for determining the distribution of exogenously administered oxytocin in an individual, the method comprising: a) administering to the individual a dose of oxytocin as described above; 13 Follow either N-labeled oxytocin peptide 13 administration of N-labeled oxytocin peptide; and b) 13 and c) imaging the cell or tissue by a non-invasive imaging technique.

[0016] In some embodiments, there is provided a method for determining the distribution of oxytocin receptors in an individual, the method comprising: a) administering to the individual a dose of oxytocin receptors as described above; 13 Follow either N-labeled oxytocin peptide 13 administration of N-labeled oxytocin peptide; and b) 13 allowing the N-labeled oxytocin peptide to bind to an oxytocin receptor; and c) in the individual. 13 and imaging the N-labeled oxytocin peptide by a non-invasive imaging technique.

[0017] In some embodiments, there is provided a method for determining the kinetics of exogenously administered oxytocin in an individual, the method comprising: a) administering to the individual a dose of oxytocin as described above; 13 Follow either N-labeled oxytocin peptide 13 and b) administering N-labeled oxytocin peptide to the individual. 13 and imaging the N-labeled oxytocin peptide over a period of time by a non-invasive imaging technique.

[0018] In some embodiments according to any of the methods described above, the non-invasive imaging technique comprises positron emission tomography imaging, hi some embodiments, the non-invasive imaging technique comprises positron emission tomography with computed tomography imaging or positron emission tomography with magnetic resonance imaging.

[0019] In some embodiments according to any of the methods described above, 13 The N-labeled oxytocin peptide is administered by craniofacial mucosal administration. 13 N-labeled oxytocin peptide is administered intranasally.

[0020] In some embodiments according to any of the methods described above, 13 The N-labeled oxytocin peptide is administered intravenously, intraarterially, intraperitoneally, intravesicularly, subcutaneously, intrathecally, intrapulmonary, intramuscularly, intratracheally, ophthalmically, transdermally, or via suppository. 13 N-labeled oxytocin peptide is administered intravenously.

[0021] In some embodiments, the above 13 Follow either N-labeled oxytocin peptide 13 Kits containing N-labeled oxytocin peptides are provided. The present invention provides, for example, the following items. (Item 1) Contains the amino acid sequence of SEQ ID NO: 1 13 N-labeled oxytocin peptide. (Item 2) One composition of the oxytocin peptide 14 N atom 13 N radionuclide, as described in item 1 13 N-labeled oxytocin peptide. (Item 3) One residue of the oxytocin peptide is 13 3. The compound according to claim 1 or 2, modified to include a moiety containing a N radionuclide. 13 N-labeled oxytocin peptide. (Item 4) One residue of the oxytocin peptide is 13 Item 3, modified to contain NH 13 N-labeled oxytocin peptide. (Item 5) Single 135. The method according to any one of items 1 to 4, comprising a N radionuclide. 13 N-labeled oxytocin peptide. (Item 6) a) a glutamine residue at position 4 (SEQ ID NO: 2), b) an asparagine residue at position 5 (SEQ ID NO: 3), or c) a glycine residue at position 9 (SEQ ID NO: 4). 13 6. The method according to any one of items 1 to 5, comprising a N radionuclide. 13 N-labeled oxytocin peptide. (Item 7) The compound of formula (IV) according to item 1 13 N-labeled oxytocin peptide. (Item 8) Contains the amino acid sequence of SEQ ID NO: 1 13 1. A method for producing an N-labeled oxytocin peptide, comprising: a) reacting a compound of formula (Ib) with diethyl cyanophosphonate (DECP) to provide a compound of formula (IIb); b) adding the compound of formula (IIb) to the gas 13 NH to provide a compound of formula (IIIb); c) deprotecting the compound of formula (IIIb) to give 13 providing N-labeled oxytocin peptide; Including, The aforementioned 13 The method, wherein the N-labeled oxytocin peptide is a compound of formula (IV): (Item 9) Item 9. The method according to item 8, wherein the reaction of the compound of formula (Ib) with DECP is carried out in the presence of dimethyl sulfoxide (DMSO), tetrahydrofuran (THF), and pentamethylpiperidine (PMP). (Item 10) 10. The method according to item 8 or 9, wherein the amount of DECP is between about 0.7 equivalents and about 1.1 equivalents. (Item 11) 11. The method of claim 9 or 10, wherein the ratio of DMSO to THF is between about 1:7 and about 1:11. (Item 12) 12. The method of any one of items 8 to 11, wherein the deprotecting comprises reacting the compound of formula (IIIb) with HCl / dioxane. (Item 13) The said provided in step c) 13 13. The method of any one of items 8 to 12, further comprising purifying the N-labeled oxytocin peptide to remove reagents, organic solvents, and precursors. (Item 14) Item 15. The method according to Item 13, wherein the purifying step comprises purification by solid phase extraction (SPE). Purification by SPE a) The above 13 The N-labeled oxytocin peptide is applied to a first hydrophobic SPE column, 13 allowing N-oxytocin and precursors to be retained on a first hydrophobic SPE column; b) applying a solution containing an aqueous ion-pairing reagent to the first hydrophobic SPE column to form the first hydrophobic SPE column; 13 allowing N-oxytocin to elute in a first eluate; c) applying the first eluate to a second hydrophobic SPE column to separate the first eluate from the second hydrophobic SPE column. 13 allowing N-oxytocin to be retained on said second hydrophobic SPE column; d) The above 13 Eluting N-oxytocin in a second eluate. Item 15. The method according to item 14, comprising: (Item 16) 16. The method of claim 15, wherein the first and / or second hydrophobic SPE column comprises a silica-based bonded phase having strong hydrophobic properties. (Item 17) 17. The method according to item 15 or 16, wherein the solution containing the aqueous ion-pairing reagent contains between about 15% and about 25% acetonitrile. (Item 18) 18. A method according to any one of items 8 to 17, 13 N-labeled oxytocin peptide. (Item 19) 1. A method for determining the distribution of exogenously administered oxytocin in an individual, comprising: a) administering to the individual a treatment according to any one of items 1 to 8 and 18 13 administering an N-labeled oxytocin peptide; b) The above 13 allowing the N-labeled oxytocin peptide to accumulate at a tissue or cell site and be imaged; c) imaging said cells or tissues by a non-invasive imaging technique; A method comprising: (Item 20) 1. A method for determining the distribution of oxytocin receptors in an individual, comprising: a) administering to the individual a treatment according to any one of items 1 to 8 and 18 13 administering an N-labeled oxytocin peptide; b) The above 13 allowing the N-labeled oxytocin peptide to bind to the oxytocin receptor; c) the individual 13 Imaging of N-labeled oxytocin peptides by non-invasive imaging techniques A method comprising: (Item 21) 1. A method for determining the kinetics of exogenously administered oxytocin in an individual, comprising: a) administering to the individual a treatment according to any one of items 1 to 8 and 18 13 administering an N-labeled oxytocin peptide; b) the individual 13 N-labeled oxytocin peptides are imaged over a period of time by non-invasive imaging techniques. A method comprising: (Item 22) 22. The method of any one of items 19 to 21, wherein the non-invasive imaging technique comprises positron emission tomography imaging. (Item 23) 23. The method of claim 22, wherein the non-invasive imaging technique comprises positron emission tomography with computed tomography imaging or positron emission tomography with magnetic resonance imaging. (Item 24) The aforementioned 13 24. The method of any one of items 19 to 23, wherein the N-labeled oxytocin peptide is administered by craniofacial mucosal administration. (Item 25) The aforementioned 13 25. The method of item 24, wherein the N-labeled oxytocin peptide is administered intranasally. (Item 26) The aforementioned 13 24. The method of any one of items 19 to 23, wherein the N-labeled oxytocin peptide is administered intravenously, intraarterially, intraperitoneally, intravesically, subcutaneously, intrathecally, intrapulmonary, intramuscularly, intratracheally, ophthalmically, transdermally, or via suppository. (Item 27) The aforementioned 13 27. The method of claim 26, wherein the N-labeled oxytocin peptide is administered intravenously. (Item 28) 19. The method according to any one of items 1 to 7 and 18. 13 Kit containing N-labeled oxytocin peptide. [Brief explanation of the drawings]

[0022] [Figure 1] FIG. 1 is a flow diagram for the synthesis and manufacture of 13N-oxytocin.

[0023] [Figure 2] FIG. 2 shows PET and MR scans of a rat after intravenous administration of oxytocin, demonstrating poor CNS penetration.

[0024] [Figure 3] Figure 3 is a superimposition of PET and MR scan images of a rat after intranasal administration of oxytocin, showing poor CNS penetration. DETAILED DESCRIPTION OF THE INVENTION

[0025] Detailed Description of the Invention For oxytocin to be therapeutically effective for a particular indication, it must be efficiently delivered to the correct target tissue. Delivery can be affected by parameters including the route of administration and formulation composition. The ability to accurately determine the spatial and temporal distribution of oxytocin administered by a route and formulation shown to be effective in treating a particular indication allows for the determination of a therapeutically effective pharmacokinetic profile for that indication. Knowledge of such a pharmacokinetic profile allows for the evaluation of new combinations of administration routes and formulations to maximize therapeutic efficacy. Furthermore, discovering the site of action of oxytocin for the treatment of various indications may provide additional therapeutic targets for investigation.

[0026] The synthesis of radiolabeled oxytocin peptides remains a challenge, hindering the use of conventional imaging techniques such as PET. 13 N-labeled oxytocin peptides and methods for their production are provided. definition

[0027] As used herein, "oxytocin peptide" refers to a substance having biological activity associated with natural oxytocin. The oxytocin peptide may be a naturally occurring endogenous peptide, a fragment, an analog, or a derivative thereof. The oxytocin peptide may also be a non-endogenous peptide, a fragment, an analog, or a derivative thereof. In one aspect, the oxytocin peptide is human oxytocin. In other aspects, the oxytocin peptide may be an analog or derivative of human oxytocin.

[0028] As used herein, "analog" or "derivative" refers to any peptide analogue of naturally occurring oxytocin in which one or more amino acids in the peptide have been substituted, deleted, or inserted. The term also refers to any peptide in which one or more amino acids (e.g., one, two, or three amino acids) have been modified, for example, by chemical modification. In general, the term covers all peptides that exhibit oxytocin activity but may, if desired, have a different potency or pharmacological profile.

[0029] The terms "therapeutic agent," "therapeutically capable agent," or "treatment" are used interchangeably and refer to a molecule or compound that confers some beneficial effect when administered to a subject. Beneficial effects include enabling a diagnostic determination; ameliorating a disease, symptom, disorder, or pathological condition; reducing or preventing the onset of a disease, symptom, disorder, or condition; and broadly combating a disease, symptom, disorder, or pathological condition.

[0030] As used herein, "treatment" or "treating" refers to an approach for obtaining beneficial or desired results, including but not limited to, therapeutic benefit. Therapeutic benefit means any improvement in, or effect on, one or more diseases, conditions, or symptoms being treated that is associated with treatment.

[0031] The term "effective amount" or "therapeutically effective amount" refers to an amount of a drug that is sufficient to produce beneficial or desired results. The therapeutically effective amount may vary depending on one or more of the subject and disease state to be treated, the subject's weight and age, the severity of the disease state, the mode of administration, etc., and can be easily determined by one skilled in the art. This term also applies to the dose that provides an image for detection by any one of the imaging methods described herein. The specific dose may vary depending on one or more of the specific drug selected, the dosing regimen to be followed, whether the drug is administered in combination with other compounds, the timing of administration, the tissue to be imaged, and the substance delivery system that carries the drug.

[0032] "Craniofacial mucosal administration" refers to delivery to mucosal surfaces of the nose, nasal passages, and nasal cavities; mucosal surfaces of the oral cavity, including the gingiva (gums), floor of the mouth, lips, tongue, lingual frenulum, and sublingual oral surfaces, including the floor of the mouth; and mucosal surfaces of the eye or periocular area, including the conjunctiva, lacrimal glands, nasolacrimal ducts, and upper and lower eyelids and ocular mucosa.

[0033] "Intranasal administration" or "intranasally administered" refers to delivery to the nose, nasal passages, or nasal cavity by spray, drops, powder, gel, film, inhalant, or other means.

[0034] The "inferior region of the nasal cavity" generally refers to the region of the nasal cavity where the middle and inferior turbinates protrude, which is the region of the nasal cavity largely innervated by the trigeminal nerve. The "superior region of the nasal cavity" is delimited by the upper third and cribriform plate region, which is innervated by the olfactory nerve.

[0035] The terms "subject," "individual," and "patient" are used interchangeably herein to refer to a vertebrate, preferably a mammal, more preferably a human. Mammals include, but are not limited to, murines, simians, humans, farm animals, sport animals, and pets. Also included are tissues, cells, and the progeny of biological entities obtained in vivo or cultured in vitro.

[0036] As used herein, the singular forms "a," "an," and "the" include plural references unless otherwise indicated.

[0037] Reference herein to "about" a value or parameter includes (and describes) embodiments directed to that value or parameter per se. For example, a statement referring to "about X" includes a statement of "X."

[0038] When a range of values ​​is stated, each intervening value between the upper and lower limits of that range, and any other stated or intervening value in that stated range, is intended to be encompassed within the disclosure. For example, if a range of 1 μg to 8 μg is stated, 2 μg, 3 μg, 4 μg, 5 μg, 6 μg, and 7 μg, as well as ranges of values ​​greater than or equal to 1 μg and ranges of values ​​less than or equal to 8 μg, are also expressly disclosed. If a range of 10 to 14% is stated, 10%, 11%, 12%, 13%, and 14% are also expressly disclosed. Furthermore, within a stated range, each smaller range between any stated or intervening value in that stated range and any other stated or intervening value is also encompassed within the disclosure. The upper and lower limits of such smaller ranges may independently be included or excluded, and each range where either, neither, or both of the smaller ranges are included is also encompassed within the disclosure, subject to the existence of any specifically excluded boundaries in the stated range. Where the stated range includes one or both of the boundaries, ranges excluding either or both of those included boundaries are also included in the disclosure.

[0039] The compositions and methods of the present invention may comprise, consist of, or consist essentially of the essential elements and limitations of the invention described herein, as well as any additional or optional ingredients, components, or limitations described herein or otherwise useful.

[0040] Unless otherwise noted, technical terms are used according to conventional usage. 13 N-oxytocin

[0041] Oxytocin was one of the first peptide hormones to be isolated and sequenced. Natural oxytocin is a nine-amino acid cyclic peptide hormone with two cysteine ​​residues forming a disulfide 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).

[0042] Methods for producing oxytocin have been described, see, for example, U.S. Patent Nos. 2,938,891 and 3,076,797. In addition, oxytocin is commercially available. Various peptide analogs and derivatives are available, and others may be contemplated for use within the present invention and may be produced and tested for biological activity according to known methods. Oxytocin analogs can include, but are not limited to, 4-threonine-1-hydroxy-deaminooxytocin, 4-serine-8-isoleucine-oxytocin, 9-deamidooxytocin, 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 a series of related compounds containing an ortho-triglyceride ethoxyphenylacetyl core, such as L-374,943.Other exemplary oxytocin analogs include 4-threonine-1-hydroxy-deaminooxytocin, 9-deamidooxytocin, an oxytocin analog containing a glycine residue in place of a glycinamide residue, 7-D-proline-oxytocin(2,4-diisoleucine)-oxytocin, an oxytocin analog with natriuretic and diuretic activity, deaminooxytocin analogs, long-acting oxytocin analogs, 1-deamino-1-monocarba-E12-[ Tyr(OMe)]-OT(dCOMOT), 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-oxytoxin (dC60), d[Lys(8)(5 / 6C-fluorescein)]VT, d[Thr(4 ),Lys(8)(5 / 6C-fluorescein)]VT, [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 oxytocin analogs in which the disulfide bridge between residues 1 and 6 is replaced with a diselenide, ditelluride, telluroseleno, tellurosulfide, or selenosulfide bond (e.g., the peptide analogs of oxytocin described in PCT Patent Application WO 2011 / 120,071, incorporated herein by reference). Peptides used within the present invention may be naturally occurring or may be obtainable by partial substitution, addition, or deletion of amino acids within a naturally occurring peptide sequence.The peptides may be chemically modified, for example, by amidation of the carboxyl terminus (—NH), use of D-amino acids in the peptide, incorporation of small nonpeptidyl moieties, and modification of the amino acids themselves (e.g., alkylation or esterification of the side chain R-groups). 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-deamidooxytocin. 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 Publication No. WO 2012 / 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 compound selected from Compound Nos. 1-65 listed in Tables 1-3 in Wisniewski, et al. J Med Chem. 2014, 57:5306-5317. In some embodiments, the oxytocin analog is selected from the group consisting of Compound No. 31 ([2-ThiMeGly7]dOT), Compound No. 47 (Carba-6-[Phe2,BuGly7]dOT), Compound No. 55 (Carba-6-[3-MeBzlGly7]dOT), and Compound No. 57 (Carba-1-[4-FBzlGly7]dOT, also known as merotocin).

[0043] An "international unit" (IU, UI, or IE) is an internationally recognized unit of activity used to quantify vitamins, hormones, and vaccines. It defines the amount of a substance that represents a unit of activity, determined using a defined biological assay to standardize preparations from multiple raw materials. Similarly, a USP unit is a distinct dosage unit established by the United States Pharmacopeia in collaboration with the Food and Drug Administration to ensure the identity, strength, quality, purity, and consistency of drug products. Generally, USP units are equivalent to international units through harmonization efforts. By convention, for oxytocin, one unit of activity is generally defined as being equivalent to approximately 2 micrograms of synthetic oxytocin peptide, or 1 mg is equivalent to 500 units (Stedman's Medical Dictionary). Therefore, as used herein, one "IU" or "international unit" of oxytocin peptide is the amount of oxytocin peptide that has the same biological activity as, or produces the same level of biological effect (e.g., contractile response in rat uterine strips) as, approximately 2 micrograms of synthetic peptide. The less active the analogue, the more material is required to achieve the same level of biological effect. Determination of drug potency is well known to those skilled in the art and may involve in vitro or in vivo assays using synthetic oxytocin as a standard. Atke and Vilhardt Acta Endocrinol 1987: 115(1):155-60; Engstrom et al. Eur J Pharmacol 1998: 355(2-3):203-10.

[0044] In some embodiments, the present invention provides 13 N-labeled oxytocin (referred to herein as " 13 In some embodiments, at least one constituent of the oxytocin peptide is provided. 14 The N atom is 13 In some embodiments, at least one amino acid of the oxytocin peptide is replaced with a N radionuclide. 13In some embodiments, the compound is modified to include a moiety that includes a N radionuclide. 13 The part containing N radionuclides is 13 NH2. In some embodiments, 13 N-oxytocin peptide is a single 13 In some embodiments, the N radionuclide 13 The N-oxytocin peptide contains a single glutamine residue at position 4. 13 In some embodiments, the N radionuclide 13 The N-oxytocin peptide contains a single asparagine residue at position 5. 13 In some embodiments, the N radionuclide 13 The N-oxytocin peptide contains a single glycine residue at position 9. 13 In some embodiments, the N radionuclide 13 N-oxytocin peptide 13 In some embodiments, the amino acid sequence comprises a glutamine, asparagine, or glycine residue modified to contain an NH. 13 The N-oxytocin peptide is a compound of formula (IV). 13 The N-oxytocin peptide may be prepared according to any of the embodiments described herein. 13 It is prepared by a process including the preparation of N-oxytocin peptide. Preparation method

[0045] In some embodiments, 13 Methods for preparing N-oxytocin peptides are provided. 13 N-oxytocin peptides can be prepared by several steps as generally described below and in more detail in the Examples below. In the process descriptions below, the symbols, when used in the formula shown, are understood to represent the groups described in connection with the formula herein.

[0046] If it is desired to obtain a specific enantiomer of a compound, this can be achieved using any conventional procedure suitable for separating or resolving enantiomers from the corresponding enantiomeric mixture. That is, for example, a mixture of enantiomers, such as a racemate, can be reacted with an appropriate chiral compound to produce a diastereomeric derivative. The diastereomeric derivatives can then be separated by any conventional means, such as crystallization, and the desired enantiomer can be recovered. In another resolution method, chiral high-performance liquid chromatography can be used to separate the racemate. Alternatively, if desired, a specific enantiomer can be obtained by using an appropriate chiral intermediate in one of the methods described.

[0047] Where it is desired to obtain a particular isomer of a compound or to otherwise purify a reaction product, chromatography (e.g., HPLC), recrystallization, and other conventional separation procedures can be employed with intermediates or final products.

[0048] Compounds of formula (IV) can be prepared according to Scheme 1, where PG1 is an amine protecting group (e.g., tert-butyloxycarbonyl (Boc), 9-fluorenylmethyloxycarbonyl (Fmoc), 2,2,2-trichloroethylformyl (Troc), carboxybenzyl, and allyloxycarbonyl), R is a phosphonate ester (e.g., diethyl phosphonate), C1-C6 alkyl (e.g., ethyl), benzyl, silyl (e.g., trimethylsilyl), or acyl (e.g., acetyl), and LG is a leaving group (e.g., OH, O-acyl, OAt, OBt, Cl, 1-imidazolyl, etc.). [ka]

[0049] Compound (I) is coupled with R-LG to give intermediate compound (II). In some embodiments, R-LG is diethyl cyanophosphonate (DECP). In some embodiments, the coupling of compound (I) with R-LG is carried out in situ in a reaction vessel. Compound (II) can be obtained by the reaction of a gas 13 In the presence of NH3, it is converted to compound (III). 13 NH3 is a liquid target (e.g., [ 16 O(p,α) 13 N) by proton irradiation 13 After making it into an aqueous NH3 solution, 13 Aqueous NH3 solution is combined with neat alkali hydroxide (e.g., NaOH) to form a gas. 13 In some embodiments, the liquid target is generated by releasing NH3. 16 O(p,α) 13 In some embodiments, the neat alkali hydroxide is NaOH. 13 The aqueous NH3 solution is transferred to a tank containing neat alkali hydroxide (e.g., NaOH) to 13 NH3 is released, for example, through Teflon tubing with a helium flow. In some embodiments, Compound (II) is 13 In the presence of NH3, at a temperature between about 0° C. and about 65° C. (e.g., about 0, 5, 10, 20, 30, 40, 50, 60, or 65° C.; including any range therebetween), the released gas is converted to compound (III). 13 The NH3 is transferred to a reaction vessel containing Compound (II). In some embodiments, the evolved gas 13 NH is manually transferred by positive pressure, for example, by use of a syringe, to the reaction vessel containing compound (II). Deprotection of the amine of compound (III) provides a compound of formula (IV).

[0050] In cases where protection of a particular reactive or incompatible group (e.g., amine or carboxylic acid) is required, it is understood that the formulae in Scheme 1 contemplate and encompass compounds in which such reactive or incompatible group is in an appropriately protected form. For a general review of protecting groups and their use, see PGM Wuts and TW Greene, Greene's Protective Groups in Organic Synthesis, 4th edition, Wiley-Interscience, New York, 2006. A preparation embodiment of the preparation method in Scheme 1 is shown in Scheme 1a, in which R 1 and R 2 are independently C1-C6 alkyl or C6-C 14 aryl, and R 1 and R 2 C1-C6 alkyl and C6-C 14 The aryl is independently optionally substituted with halogen, cyano, or C1-C6 alkyl, PG1 is an amine protecting group (e.g., tert-butyloxycarbonyl (Boc), 9-fluorenylmethyloxycarbonyl (Fmoc), 2,2,2-trichloroethylformyl (Troc), carboxybenzyl, and allyloxycarbonyl), and LG is a leaving group (e.g., OH, O-acyl, OAt, OBt, Cl, 1-imidazolyl, etc.). [ka]

[0051] Compound (Ia) is reacted with the phosphonate ester compound LG-P(=O)(OR 1 )(OR 2 ) (e.g., diethyl cyanophosphonate) to provide intermediate compound (IIa). In some embodiments, the LG-P(=O)(OR 1 )(OR 2 The coupling with the compound (IIa) is carried out in situ in a reaction vessel. 13In the presence of NH3, it is converted to compound (IIIa). 13 NH3 is a liquid target (e.g., [ 16 O(p,α) 13 N) by proton irradiation 13 After making it into an aqueous NH3 solution, 13 Aqueous NH3 solution is combined with neat alkali hydroxide (e.g., NaOH) to form a gas. 13 In some embodiments, the liquid target is generated by releasing NH3. 16 O(p,α) 13 In some embodiments, the neat alkali hydroxide is NaOH. 13 The aqueous NH3 solution is transferred to a tank containing neat alkali hydroxide (e.g., NaOH) to 13 NH3 is released, for example, through Teflon tubing with a helium flow. In some embodiments, compound (IIa) is 13 In the presence of NH3, at a temperature between about 0° C. and about 65° C. (e.g., about 0, 5, 10, 20, 30, 40, 50, 60, or 65° C.; including any range therebetween), the released gas is converted to compound (IIIa). 13 NH3 is transferred to a reaction vessel containing compound (IIa). In some embodiments, the evolved gas 13 NH is manually transferred by positive pressure, for example, by use of a syringe, to the reaction vessel containing compound (IIa). Deprotection of the amine of compound (IIIa) provides a compound of formula (IV).

[0052] Another preparation embodiment of the preparation method in Scheme 1 is shown in Scheme 1b. [ka]

[0053] Compound (Ib) is coupled with diethyl cyanophosphonate (DECP) to provide intermediate compound (IIb). In some embodiments, compound (Ib) is coupled with DECP in the presence of dimethyl sulfoxide (DMSO), tetrahydrofuran (THF), and pentamethylpiperidine (PMP). In some embodiments, the amount of DECP in the coupling reaction is between about 0.7 equivalents and about 1.1 equivalents (e.g., about 0.7, 0.8, 0.9, 1.0, and 1.1 equivalents). In some embodiments, the amount of DECP in the coupling reaction is about 0.9 equivalents. In some embodiments, the ratio of DMSO to THF in the coupling reaction is between about 1:7 and about 1:11 (e.g., about 1:7, 1:8, 1:9, 1:10, and 1:11). In some embodiments, the ratio of DMSO to THF in the coupling reaction is about 1:9. In some embodiments, the coupling of compound (Ib) with DECP is carried out at a temperature between about 0°C and about 65°C (e.g., about 0, 5, 10, 20, 30, 40, 50, 60, or 65°C; including any range therebetween). In some embodiments, the coupling of compound (Ib) with DEPC is carried out in situ in a reaction vessel. In some embodiments, compound (Ib) is coupled with DECP in the presence of DMF / dioxane, DMF / THF, or DMSO / dioxane. Compound (IIb) can be prepared in a gas phase. 13 In the presence of NH3, it is converted to compound (IIIb). 13 NH3 is a liquid target (e.g., [ 16 O(p,α) 13 N) by proton irradiation 13 After making it into an aqueous NH3 solution, 13 Aqueous NH3 solution is combined with neat alkali hydroxide (e.g., NaOH) to form a gas. 13 In some embodiments, the liquid target is generated by releasing NH3. 16 O(p,α) 13 In some embodiments, the neat alkali hydroxide is NaOH. 13The aqueous NH3 solution is transferred to a tank containing neat alkali hydroxide (e.g., NaOH) to 13 NH3 is released, for example, through Teflon tubing with a helium flow. In some embodiments, compound (IIb) is 13 In the presence of NH3, at a temperature between about 0° C. and about 65° C. (e.g., about 0, 5, 10, 20, 30, 40, 50, 60, or 65° C.; including any range therebetween), the released gas is converted to compound (IIIb). 13 NH3 is transferred to a reaction vessel containing compound (IIb). In some embodiments, the evolved gas 13 NH3 is manually transferred by positive pressure, e.g., by use of a syringe, into a reaction vessel containing compound (IIb). Deprotection of the amine of compound (IIIb) provides a compound of formula (IV). In some embodiments, compound (IIIb) is deprotected in the presence of HCl and dioxane to provide compound (IV). In some embodiments, the HCl is at a concentration of between about 3.5 M and about 4.5 M (e.g., about any of 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, or 4.5 M). In some embodiments, the deprotection is carried out at a temperature between about 45°C and about 55°C (e.g., about any of 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, or 55°C). In some embodiments, the deprotection is carried out at about 50°C. In some embodiments, the deprotection is carried out for a period of between about 1 minute and about 5 minutes (e.g., about any of 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, or 5 minutes). In some embodiments, the deprotection is carried out for about 2.5 minutes. In some embodiments, compound (IIIb) is deprotected in the presence of HCl in diethyl ether (e.g., 2M HCl), HCl in THF / dioxane (e.g., 1M HCl), or HCl in MeOH (e.g., 1M HCl) to provide compound (IV).

[0054] In some embodiments, deprotection of compound (III), (IIIa), or (IIIb) provides a crude preparation of a compound of formula (IV). In some embodiments, the crude preparation of compound (IV) is purified to remove reagents, organic solvents, and / or precursors. In some embodiments, purification is by high performance liquid chromatography (HPLC) or solid phase extraction (SPE). In some embodiments, purification is performed in one step. In some embodiments, purification is performed in multiple steps. For example, in some embodiments, purification includes: a) applying the crude preparation of compound (IV) to a first hydrophobic SPE column; 13 allowing N-oxytocin and precursors of the crude preparation to be retained on a first hydrophobic SPE column; and b) applying a solution comprising an aqueous ion-pairing reagent to the first hydrophobic SPE column, 13 allowing N-oxytocin to elute in a first eluate; and c) applying the first eluate to a second hydrophobic SPE column to obtain a second hydrophobic SPE column. 13 allowing N-oxytocin to be retained on a second hydrophobic SPE column; and d) 13 and eluting N-oxytocin in a second eluate. In some embodiments, the first and / or second hydrophobic SPE column comprises a silica-based bonded phase with strong hydrophobicity, such as a Sep-Pak C18 cartridge (Waters). In some embodiments, the crude preparation is diluted before being applied to the first hydrophobic SPE column. In some embodiments, the solution containing the aqueous ion-pairing reagent comprises acetonitrile. In some embodiments, the solution containing the aqueous ion-pairing reagent comprises acetonitrile at a concentration of between about 15% and about 25% (e.g., about any of 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25%). In some embodiments, the first eluate is diluted before being applied to the second hydrophobic SPE column. How to use Imaging

[0055] In some embodiments, a method of in vivo imaging of exogenously administered oxytocin in a subject is provided, the method comprising: (a) administering to the subject a vasoconstrictor according to any of the embodiments described herein;13 (b) administering N-labeled oxytocin peptide; 13 and (c) imaging the cells or tissue by a non-invasive imaging technique. The non-invasive imaging technique may be positron emission tomography imaging, or positron emission tomography with computed tomography imaging, or positron emission tomography with magnetic resonance imaging. In some embodiments, 13 The N-oxytocin peptide is a compound of formula (IV). 13 N-labeled oxytocin is administered by craniofacial mucosal administration (e.g., nasal, buccal, or sublingual administration), for example, to image central nervous system (CNS) tissue. 13 N-labeled oxytocin is administered intranasally. 13 N-labeled oxytocin is administered intravenously, intraarterially, intraperitoneally, intravesically, subcutaneously, intrathecally, intrapulmonary, intramuscularly, intratracheally, ophthalmically (e.g., intraocularly), transdermally, or via suppository, for example, to image peripheral tissues. 13 N-labeled oxytocin is administered intravenously.

[0056] 13 In some embodiments according to any of the methods described herein comprising administration of an N-labeled oxytocin peptide, 13 The N-labeled oxytocin peptide is co-administered with a non-radiolabeled oxytocin peptide. 13 The N-labeled oxytocin peptide and the non-radiolabeled oxytocin peptide are contained in a single formulation or composition. 13 The N-labeled oxytocin peptide and the non-radiolabeled oxytocin peptide are contained in separate formulations or compositions. In some embodiments, the method further comprises administering a quantity (e.g., an effective amount) of a composition comprising non-radiolabeled oxytocin. In some embodiments, the administration of non-radiolabeled oxytocin comprises: 13Prior to administration of N-labeled oxytocin peptide, e.g. 13 In some embodiments, the administration of non-radiolabeled oxytocin is performed at least about 10 minutes (e.g., at least about 10 minutes, 20 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 3 hours, 4 hours, or more; including any ranges therebetween) prior to administration of the N-labeled oxytocin peptide. 13 After administration of N-labeled oxytocin peptide, e.g. 13 The administration of the N-labeled oxytocin peptide is performed at least about 10 minutes (e.g., at least about 10 minutes, 20 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 3 hours, 4 hours, or more; including any ranges therebetween). In some embodiments, the administration of non-radiolabeled oxytocin is performed at least about 10 minutes (e.g., at least about 10 minutes, 20 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 3 hours, 4 hours, or more; including any ranges therebetween). 13 After administration of N-labeled oxytocin peptide, e.g. 13 In some embodiments, the method is performed at least about 10 minutes (e.g., at least about 10 minutes, 20 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 3 hours, 4 hours, or more; including any ranges therebetween) after administration of the N-labeled oxytocin peptide, and the method further comprises repeating steps (a), (b), and (c) after administration of non-radiolabeled oxytocin, e.g., at least about 10 minutes (e.g., at least about 10 minutes, 20 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 3 hours, 4 hours, or more; including any ranges therebetween) after administration of non-radiolabeled oxytocin. In some embodiments, the method is performed at least about 10 minutes (e.g., at least about 10 minutes, 20 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 3 hours, 4 hours, or more; including any ranges therebetween) after administration of non-radiolabeled oxytocin. 13 When N-labeled oxytocin peptide is administered, two doses 13 The time interval between administrations of N-labeled oxytocin peptide is at least about 20 minutes (e.g., at least about 20 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 3 hours, 4 hours, or longer; including any ranges therebetween). In some embodiments, the method further comprises administering one or more additional doses (e.g., effective doses) of non-radiolabeled oxytocin. In some embodiments, the non-radiolabeled oxytocin 13When administered separately from N-labeled oxytocin, non-radiolabeled oxytocin 13 In some embodiments, non-radiolabeled oxytocin is administered by the same route as N-labeled oxytocin. 13 When administered separately from N-labeled oxytocin, non-radiolabeled oxytocin 13 It is administered by a different route than N-labeled oxytocin. In some embodiments, non-radiolabeled oxytocin is administered via craniofacial mucosal administration (e.g., nasal, buccal, or sublingual administration). In some embodiments, non-radiolabeled oxytocin is administered intranasally. In some embodiments, non-radiolabeled oxytocin is administered intravenously, intraarterially, intraperitoneally, intravesicularly, subcutaneously, intrathecally, intrapulmonary, intramuscularly, intratracheally, ophthalmically (e.g., intraocularly), transdermally, or via suppository. In some embodiments, non-radiolabeled oxytocin is administered intravenously.

[0057] 13 In some embodiments according to any of the methods described herein comprising administration of an N-labeled oxytocin peptide, 13 The N-labeled oxytocin peptide is co-administered with another agent. In some embodiments, the other agent affects the distribution, kinetics, and / or pharmacodynamics of oxytocin. In some embodiments, the other agent is a divalent metal cation (e.g., Mg 2+ , Ca 2+ , Zn 2+ , or Cu 2+ In some embodiments, the divalent metal cation is Mg 2+ In some embodiments, the other agent is vasopressin. 13 The N-labeled oxytocin peptide and the other agent are contained in a single formulation or composition. For example, in some embodiments, the method comprises: 13 N-labeled oxytocin peptide and divalent metal cations (e.g., Mg 2+ , Ca 2+ , Zn 2+ , or Cu 2+ In some embodiments, the administration of a formulation or composition comprising: 13The N-labeled oxytocin peptide and the other agent are contained in separate formulations or compositions. For example, in some embodiments, the method comprises: 13 Administration of a formulation or composition containing N-labeled oxytocin peptide and a divalent metal cation (e.g., Mg 2+ , Ca 2+ , Zn 2+ , or Cu 2+ ) or vasopressin. In some embodiments, the other agent is 13 In some embodiments, the other agent and 13 In some embodiments, the other agent is: 13 In some embodiments, the other agent is administered before the N-labeled oxytocin peptide. 13 In some embodiments, the other agent is administered after the N-labeled oxytocin peptide. 13 Other drugs, when administered separately from N-labeled oxytocin, 13 In some embodiments, the other agent is administered by the same route as N-labeled oxytocin. 13 Other drugs, when administered separately from N-labeled oxytocin, 13 The other agent is administered by a different route than the N-labeled oxytocin. In some embodiments, the other agent is administered by craniofacial mucosal administration (e.g., nasal, buccal, or sublingual administration). In some embodiments, the other agent is administered intranasally. In some embodiments, the other agent is administered intravenously, intraarterially, intraperitoneally, intravesicularly, subcutaneously, intrathecally, intrapulmonary, intramuscularly, intratracheally, ophthalmologically (e.g., intraocularly), transdermally, or via suppository. In some embodiments, the other agent is administered intravenously.

[0058] In some embodiments, a method of imaging exogenously administered oxytocin in a subject by emission tomography is provided, the method comprising: (a) administering to the subject a radiofrequency ablation therapy according to any of the embodiments described herein; 13(b) detecting gamma rays emitted from the subject using a plurality of detectors and transmitting signals corresponding to the detected gamma rays; and (c) reconstructing a series of medical images of a region of interest in the subject from the signals. 13 The N-oxytocin peptide is a compound of formula (IV). 13 The N-oxytocin peptide is administered by craniofacial mucosal administration (e.g., intranasal administration). 13 The N-oxytocin peptide may be administered intravenously, intraarterially, intraperitoneally, intravesically, subcutaneously, intrathecally, intrapulmonary, intramuscularly, intratracheally, ophthalmically (eg, intraocularly), transdermally, or via suppository.

[0059] In some embodiments, the method is according to any of the embodiments described herein. 13 Imaging methods are provided that include obtaining an image of a human patient to which a detectable amount of N-labeled oxytocin peptide has been administered. The method may include obtaining an image (e.g., a brain image) of the patient using positron emission tomography imaging, or positron emission tomography with computed tomography imaging, or positron emission tomography with magnetic resonance imaging. 13 Detectable amounts of N-labeled oxytocin peptide in cells or tissues 13 The amount is sufficient to allow the accumulation of N-labeled oxytocin peptide to be detected by medical imaging techniques. 13 The N-oxytocin peptide is a compound of formula (IV). 13 The N-oxytocin peptide is administered by craniofacial mucosal administration (e.g., intranasal administration). 13 The N-oxytocin peptide may be administered intravenously, intraarterially, intraperitoneally, intravesically, subcutaneously, intrathecally, intrapulmonary, intramuscularly, intratracheally, ophthalmically (eg, intraocularly), transdermally, or via suppository.

[0060] In some embodiments, a method for determining the distribution of exogenously administered oxytocin in a subject is provided, comprising: (a) administering to the subject a dose of oxytocin according to any of the embodiments described herein; 13 (b) administering N-labeled oxytocin peptide; 13 (c) allowing the N-labeled oxytocin peptide to accumulate at the tissue or cell site to be imaged; and (c) imaging the cell or tissue by a non-invasive imaging technique. The non-invasive imaging technique may be positron emission tomography imaging, or positron emission tomography with computed tomography imaging, or positron emission tomography with magnetic resonance imaging. In some embodiments, 13 The N-oxytocin peptide is a compound of formula (IV). 13 The N-oxytocin peptide is administered by craniofacial mucosal administration (e.g., intranasal administration). 13 The N-oxytocin peptide may be administered intravenously, intraarterially, intraperitoneally, intravesically, subcutaneously, intrathecally, intrapulmonary, intramuscularly, intratracheally, ophthalmically (eg, intraocularly), transdermally, or via suppository.

[0061] In some embodiments, a method for determining the distribution of oxytocin receptors in a subject is provided. In some embodiments, the method comprises: (a) administering to the subject a dose of oxytocin receptors according to any of the embodiments described herein; 13 (b) administering N-labeled oxytocin peptide; 13 (c) allowing the N-labeled oxytocin peptide to bind to an oxytocin receptor in an individual; 13 and imaging the N-labeled oxytocin peptide by a non-invasive imaging technique. The non-invasive imaging technique may be positron emission tomography imaging, or positron emission tomography with computed tomography imaging, or positron emission tomography with magnetic resonance imaging. In some embodiments, 13 The N-oxytocin peptide is a compound of formula (IV). 13The N-oxytocin peptide is administered by craniofacial mucosal administration (e.g., intranasal administration). 13 The N-oxytocin peptide may be administered intravenously, intraarterially, intraperitoneally, intravesically, subcutaneously, intrathecally, intrapulmonary, intramuscularly, intratracheally, ophthalmically (eg, intraocularly), transdermally, or via suppository.

[0062] In some embodiments, a method for determining the kinetics of exogenously administered oxytocin in an individual is provided, comprising: (a) administering to the subject a dose of oxytocin according to any of the embodiments described herein; 13 (b) administering N-labeled oxytocin peptide to an individual 13 and imaging the N-labeled oxytocin peptide over a period of time by a non-invasive imaging technique, which may be positron emission tomography imaging, or positron emission tomography with computed tomography imaging, or positron emission tomography with magnetic resonance imaging. 13 The N-oxytocin peptide is a compound of formula (IV). 13 The N-oxytocin peptide is administered by craniofacial mucosal administration (e.g., intranasal administration). 13 The N-oxytocin peptide may be administered intravenously, intraarterially, intraperitoneally, intravesically, subcutaneously, intrathecally, intrapulmonary, intramuscularly, intratracheally, ophthalmically (eg, intraocularly), transdermally, or via suppository. Emission tomography

[0063] In some embodiments, the methods described herein comprise: 13 In some embodiments, an emission tomography system is used to acquire a series of medical images of a subject during an imaging process using N-labeled oxytocin peptide as a radiotracer. In some embodiments, the system detects gamma rays emitted by the subject and transmits them to the subject. 13The method includes a plurality of detectors configured to be positioned around the subject to acquire gamma rays over a predetermined period of time relative to administration of an N-labeled oxytocin peptide and transmit signals corresponding to the acquired gamma rays, a data processing system configured to receive the signals from the plurality of detectors, and a reconstruction system configured to receive the signals from the data processing system and reconstruct a series of medical images of the subject therefrom. 13 N-labeled oxytocin peptides as described herein 13 N-labeled oxytocin peptide. treatment

[0064] In some embodiments, methods are provided for treating disorders or conditions, including psychiatric, neuropsychiatric, and psychological disorders, inflammatory conditions, and pain, comprising administering to an individual in need thereof an effective dose of an oxytocin peptide formulation, wherein administration of the oxytocin peptide formulation results in a distribution and / or kinetics of the oxytocin peptide in the individual that is favorable for treating the disorder or condition. In some embodiments, the distribution and / or kinetics of the oxytocin peptide in the individual results in the individual: 13 Contains N-labeled oxytocin 13 administering an N-oxytocin peptide formulation or composition to an individual 13 The N-labeled oxytocin peptide is revealed by imaging, for example, by PET, according to any of the embodiments described herein. In some embodiments, the oxytocin peptide formulation further comprises non-radiolabeled oxytocin. In some embodiments, the oxytocin peptide formulation comprises an effective amount of oxytocin ( 13 In some embodiments, the oxytocin peptide formulation comprises a) an effective amount of non-radiolabeled oxytocin, and b) an amount suitable for use as an imaging tracer, e.g., a PET tracer. 13and N-labeled oxytocin. In some embodiments, the oxytocin peptide formulation is administered via craniofacial mucosal administration (e.g., intranasal administration). In some embodiments, the oxytocin peptide formulation is administered intravenously, intraarterially, intraperitoneally, intravesicularly, subcutaneously, intrathecally, intrapulmonary, intramuscularly, intratracheally, ophthalmically (e.g., intraocularly), transdermally, or via suppository. In some embodiments, the neuropsychiatric disorder includes autism spectrum disorder, a disorder exhibiting one or more symptoms associated with autism spectrum disorder, a social and communication disorder, and an anxiety disorder. In some embodiments, the pain includes orofacial and craniofacial pain (e.g., headache), neck pain (e.g., occipital neuralgia), and pain in the upper limbs.

[0065] For example, in some embodiments, a method of treating a pain and / or inflammatory condition is provided, comprising administering to an individual in need thereof an effective dose of an oxytocin peptide formulation, wherein administration of the oxytocin peptide formulation results in distribution of the oxytocin peptide to the nasal mucosa innervated by the trigeminal nerve. In some embodiments, distribution of the oxytocin peptide in the individual is achieved by inducing the individual to: 13 Contains N-labeled oxytocin 13 administering an N-oxytocin peptide formulation or composition to an individual 13 The N-labeled oxytocin peptide is revealed by imaging, for example, by PET, according to any of the embodiments described herein. In some embodiments, the oxytocin peptide formulation is administered via craniofacial mucosal administration (e.g., intranasal administration). In some embodiments, the oxytocin peptide formulation is administered intravenously. In some embodiments, the pain includes orofacial and craniofacial pain (e.g., headache), neck pain (e.g., occipital neuralgia), and upper extremity pain.

[0066] In some embodiments, a method of treating a psychiatric, neuropsychiatric, or psychological disorder is provided, comprising administering to an individual in need thereof an effective dose of an oxytocin peptide formulation, wherein administration of the oxytocin peptide formulation results in distribution of oxytocin peptide to the nasal mucosa innervated by the olfactory nerve (e.g., the upper most mucosa). In some embodiments, distribution of oxytocin peptide in the individual results in the individual receiving detectable amounts of oxytocin peptide. 13 oxytocin peptide preparations or compositions containing N-labeled oxytocin are administered to individuals, 13 The N-labeled oxytocin peptide is revealed by imaging, for example, by PET, according to any of the embodiments described herein. In some embodiments, the oxytocin peptide formulation is administered via craniofacial mucosal administration (e.g., intranasal administration). In some embodiments, the oxytocin peptide formulation is administered intravenously. In some embodiments, the neuropsychiatric disorder includes autism spectrum disorder, a disorder exhibiting one or more symptoms associated with autism spectrum disorder, social and communication disorder, and anxiety disorder.

[0067] In some embodiments, a method of evaluating an oxytocin peptide formulation for the treatment of a disorder or condition, including psychiatric, neuropsychiatric, and psychological disorders, inflammatory conditions, and pain, in an individual, comprising: a) administering to an individual an oxytocin peptide formulation containing an oxytocin peptide formulation to an individual; 13 administering to the individual an oxytocin peptide formulation comprising N-labeled oxytocin; and b) administering to the individual an oxytocin peptide formulation comprising N-labeled oxytocin. 13 determining the distribution and / or kinetics of N-labeled oxytocin according to any of the embodiments described herein; and c) in the individual. 13 and evaluating the oxytocin peptide formulation for the treatment of a disorder or condition based on the favorable distribution and / or kinetics of N-labeled oxytocin. In some embodiments, the oxytocin peptide formulation further comprises non-radiolabeled oxytocin. In some embodiments, the oxytocin peptide formulation contains an effective amount of oxytocin ( 13In some embodiments, the oxytocin peptide formulation comprises a) an effective amount of non-radiolabeled oxytocin, and b) an amount suitable for use as an imaging tracer, e.g., a PET tracer. 13 and N-labeled oxytocin. In some embodiments, the oxytocin peptide formulation is administered via craniofacial mucosal administration (e.g., intranasal administration). In some embodiments, the oxytocin peptide formulation is administered intravenously, intraarterially, intraperitoneally, intravesicularly, subcutaneously, intrathecally, intrapulmonary, intramuscularly, intratracheally, ophthalmically (e.g., intraocularly), transdermally, or via suppository.

[0068] In some embodiments, a method of evaluating an oxytocin peptide formulation for the treatment of a disorder or condition, including psychiatric, neuropsychiatric, and psychological disorders, inflammatory conditions, and pain, comprising: a) administering to a subject a therapeutically effective amount of oxytocin peptide formulation to a subject; 13 administering to a plurality of individuals an oxytocin peptide formulation comprising N-labeled oxytocin; and b) administering to a plurality of individuals an oxytocin peptide formulation comprising N-labeled oxytocin. 13 determining the distribution and / or kinetics of N-labeled oxytocin according to any of the embodiments described herein; and c) determining the distribution and / or kinetics of N-labeled oxytocin in a plurality of individuals. 13 and evaluating the oxytocin peptide formulation for the treatment of a disorder or condition based on the favorable distribution and / or kinetics of N-labeled oxytocin. In some embodiments, the oxytocin peptide formulation further comprises non-radiolabeled oxytocin. In some embodiments, the oxytocin peptide formulation contains an effective amount of oxytocin ( 13 In some embodiments, the oxytocin peptide formulation comprises a) an effective amount of non-radiolabeled oxytocin, and b) an amount suitable for use as an imaging tracer, e.g., a PET tracer. 13and N-labeled oxytocin. In some embodiments, the oxytocin peptide formulation is administered via craniofacial mucosal administration (e.g., intranasal administration). In some embodiments, the oxytocin peptide formulation is administered intravenously, intraarterially, intraperitoneally, intravesicularly, subcutaneously, intrathecally, intrapulmonary, intramuscularly, intratracheally, ophthalmically (e.g., intraocularly), transdermally, or via suppository.

[0069] In some embodiments, a method of selecting an oxytocin peptide formulation for treating a disorder or condition, including psychiatric, neuropsychiatric, and psychological disorders, inflammatory conditions, and pain, in an individual is provided, comprising: a) selecting an oxytocin peptide formulation for treating a disorder or condition, including psychiatric, neuropsychiatric, and psychological disorders, inflammatory conditions, and pain, in an individual; 13 administering to an individual a plurality of oxytocin peptide formulations, each comprising N-labeled oxytocin; and b) determining whether or not a particular oxytocin peptide formulation is effective in the individual for each of the plurality of oxytocin peptide formulations. 13 determining the distribution and / or kinetics of N-labeled oxytocin according to any of the embodiments described herein; and c) in the individual. 13 and selecting an oxytocin peptide formulation in which the distribution and / or kinetics of N-labeled oxytocin are most favorable for treating the disorder or condition. In some embodiments, the oxytocin peptide formulation further comprises non-radiolabeled oxytocin. In some embodiments, the oxytocin peptide formulation contains an effective amount of oxytocin (N-labeled oxytocin) to treat the disorder or condition. 13 In some embodiments, the oxytocin peptide formulation comprises a) an effective amount of non-radiolabeled oxytocin, and b) an amount suitable for use as an imaging tracer, e.g., a PET tracer. 13 and N-labeled oxytocin. In some embodiments, the oxytocin peptide formulation is administered via craniofacial mucosal administration (e.g., intranasal administration). In some embodiments, the oxytocin peptide formulation is administered intravenously, intraarterially, intraperitoneally, intravesicularly, subcutaneously, intrathecally, intrapulmonary, intramuscularly, intratracheally, ophthalmically (e.g., intraocularly), transdermally, or via suppository.

[0070] In some embodiments, a method of selecting an oxytocin peptide formulation for treating a disorder or condition, including psychiatric, neuropsychiatric, and psychological disorders, inflammatory conditions, and pain, is provided, comprising: a) administering to the subject a therapeutically effective amount of an oxytocin peptide formulation ... 13 administering to a plurality of individuals a plurality of oxytocin peptide formulations, each comprising N-labeled oxytocin; and b) determining whether or not a plurality of individuals, for each of the plurality of oxytocin peptide formulations, 13 determining the distribution and / or kinetics of N-labeled oxytocin according to any of the embodiments described herein; and c) determining the distribution and / or kinetics of N-labeled oxytocin in a plurality of individuals. 13 and selecting an oxytocin peptide formulation in which the distribution and / or kinetics of N-labeled oxytocin are most favorable for treating the disorder or condition. In some embodiments, the oxytocin peptide formulation further comprises non-radiolabeled oxytocin. In some embodiments, the oxytocin peptide formulation contains an effective amount of oxytocin (N-labeled oxytocin) to treat the disorder or condition. 13 In some embodiments, the oxytocin peptide formulation comprises a) an effective amount of non-radiolabeled oxytocin, and b) an amount suitable for use as an imaging tracer, e.g., a PET tracer. 13 and N-labeled oxytocin. In some embodiments, the oxytocin peptide formulation is administered via craniofacial mucosal administration (e.g., intranasal administration). In some embodiments, the oxytocin peptide formulation is administered intravenously, intraarterially, intraperitoneally, intravesicularly, subcutaneously, intrathecally, intrapulmonary, intramuscularly, intratracheally, ophthalmically (e.g., intraocularly), transdermally, or via suppository.

[0071] In some embodiments, there is provided a method for evaluating a therapy comprising administration of an oxytocin peptide formulation for treating a disorder or condition, including psychiatric, neuropsychiatric, and psychological disorders, inflammatory conditions, and pain, in an individual, comprising: a) administering the oxytocin peptide formulation to a subject; 13 b) treating the individual with a therapy comprising N-labeled oxytocin; and 13determining the distribution and / or kinetics of N-labeled oxytocin according to any of the embodiments described herein; and c) in the individual. 13 and evaluating a therapy for treating a disorder or condition based on favorable distribution and / or kinetics of N-labeled oxytocin. In some embodiments, the oxytocin peptide formulation further comprises non-radiolabeled oxytocin. In some embodiments, the oxytocin peptide formulation comprises an effective amount of oxytocin ( 13 In some embodiments, the oxytocin peptide formulation comprises a) an effective amount of non-radiolabeled oxytocin, and b) an amount suitable for use as an imaging tracer, e.g., a PET tracer. 13 N-labeled oxytocin. The therapy may include an oxytocin peptide formulation according to any of the formulations described herein, such as those described in the section entitled "Oxytocin Peptide Formulations," administered by any of the administration methods described herein, such as those described in the section entitled "Oxytocin Administration."

[0072] In some embodiments, there is provided a method for evaluating a therapy comprising administration of an oxytocin peptide formulation for treating a disorder or condition, including psychiatric, neuropsychiatric, and psychological disorders, inflammatory conditions, and pain, comprising: a) administering an oxytocin peptide formulation to a subject; 13 b) treating a plurality of individuals with a therapy comprising N-labeled oxytocin; and 13 determining the distribution and / or kinetics of N-labeled oxytocin according to any of the embodiments described herein; and c) determining the distribution and / or kinetics of N-labeled oxytocin in a plurality of individuals. 13 and evaluating a therapy for treating a disorder or condition based on favorable distribution and / or kinetics of N-labeled oxytocin. In some embodiments, the oxytocin peptide formulation further comprises non-radiolabeled oxytocin. In some embodiments, the oxytocin peptide formulation comprises an effective amount of oxytocin ( 13In some embodiments, the oxytocin peptide formulation comprises a) an effective amount of non-radiolabeled oxytocin, and b) an amount suitable for use as an imaging tracer, e.g., a PET tracer. 13 N-labeled oxytocin. The therapy may include an oxytocin peptide formulation according to any of the formulations described herein, such as those described in the section entitled "Oxytocin Peptide Formulations," administered by any of the administration methods described herein, such as those described in the section entitled "Oxytocin Administration."

[0073] In some embodiments, a method for selecting a therapy comprising administration of an oxytocin peptide formulation for treating a disorder or condition, including psychiatric, neuropsychiatric, and psychological disorders, inflammatory conditions, and pain, in an individual, comprising: a) administering an oxytocin peptide formulation to a patient; 13 treating the individual with multiple therapies including administration of an oxytocin peptide formulation comprising N-labeled oxytocin; and b) determining, for each of the multiple therapies, whether or not the individual has a phenotype different from that of the N-labeled oxytocin. 13 determining the distribution and / or kinetics of N-labeled oxytocin according to any of the embodiments described herein; and c) in the individual. 13 and selecting a therapy in which the distribution and / or kinetics of N-labeled oxytocin are most favorable for treating the disorder or condition. In some embodiments, the oxytocin peptide formulation further comprises non-radiolabeled oxytocin. In some embodiments, the oxytocin peptide formulation comprises an effective amount of oxytocin (N-labeled oxytocin) to treat the disorder or condition. 13 In some embodiments, the oxytocin peptide formulation comprises a) an effective amount of non-radiolabeled oxytocin, and b) an amount suitable for use as an imaging tracer, e.g., a PET tracer. 13N-labeled oxytocin. The therapy may include an oxytocin peptide formulation according to any of the formulations described herein, such as those described in the section entitled "Oxytocin Peptide Formulations," administered by any of the administration methods described herein, such as those described in the section entitled "Oxytocin Administration."

[0074] In some embodiments, a method for selecting a therapy for treating a disorder or condition, including psychiatric, neuropsychiatric, and psychological disorders, inflammatory conditions, and pain, comprising administering an oxytocin peptide formulation, comprises: a) administering an oxytocin peptide formulation to a patient in need thereof; 13 treating a plurality of individuals with a plurality of therapies comprising administration of an oxytocin peptide formulation comprising N-labeled oxytocin; and b) determining, for each of the plurality of therapies, the level of oxytocin in each of the plurality of individuals. 13 determining the distribution and / or kinetics of N-labeled oxytocin according to any of the embodiments described herein; and c) determining the distribution and / or kinetics of N-labeled oxytocin in a plurality of individuals. 13 and selecting a therapy in which the distribution and / or kinetics of N-labeled oxytocin are most favorable for treating the disorder or condition. In some embodiments, the oxytocin peptide formulation further comprises non-radiolabeled oxytocin. In some embodiments, the oxytocin peptide formulation comprises an effective amount of oxytocin (N-labeled oxytocin) to treat the disorder or condition. 13 In some embodiments, the oxytocin peptide formulation comprises a) an effective amount of non-radiolabeled oxytocin, and b) an amount suitable for use as an imaging tracer, e.g., a PET tracer. 13 N-labeled oxytocin. The therapy may include an oxytocin peptide formulation according to any of the formulations described herein, such as those described in the section entitled "Oxytocin Peptide Formulations," administered by any of the administration methods described herein, such as those described in the section entitled "Oxytocin Administration." Diseases and Conditions

[0075] The terms "autism spectrum disorder (ASD)" or "autism" refer to a group of complex disorders of brain development. These disorders are characterized, to varying degrees, by difficulties with social interaction, verbal and nonverbal communication, and repetitive behaviors. With the May 2013 publication of the fifth edition of the Diagnostic and Statistical Manual of Mental Disorders (DSM-5), all autistic disorders were combined into one diagnosis: ASD. Previously, such disorders, including autistic disorder, childhood disintegrative disorder, pervasive developmental disorder not otherwise specified (PDD-NOS), and Asperger's syndrome, were recognized as distinct subtypes. See http: / / www.autismspeaks.org / what-autism. Those skilled in the art will recognize that the symptoms of autism spectrum disorder share some overlap with many other psychiatric disorders. 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.

[0076] Autism spectrum disorder (ASD) is characterized by social interaction difficulties, communication challenges, and a tendency to engage in repetitive behaviors. However, symptoms and their severity vary widely across these three core ranges. ASD may also be associated with intellectual disability, motor coordination and attention difficulties, and physical health problems such as sleep and gastrointestinal disorders. ASD may also be associated with psychiatric symptoms, including anxiety and depression. See, for example, Kim et al., Autism 2000, 4(2):117-132.

[0077] Oxytocin is known to treat several conditions, including anxiety and social and communication disorders in autism spectrum disorder. However, the effectiveness of oxytocin in treating social and communication disorders in autism spectrum disorder has been observed to vary greatly among patients. Variations in oxytocin's receptor availability and receptor affinity may contribute to the variability in effectiveness. Clinical efforts to use commercially available oxytocin preparations (e.g., Syntocinon®) to treat ASD have been frustrated by insufficient efficacy and poor tolerability. Currently available oxytocin preparations are weakly potent and contain large amounts of drug, so when administered via nasal spray, the amount of drug absorbed is insufficient to be effective. In some embodiments, the present invention provides a method for evaluating oxytocin peptide therapy for treating autism spectrum disorder, a disorder exhibiting one or more symptoms associated with autism spectrum disorder, a social and communication disorder, or an anxiety disorder.

[0078] In some embodiments according to any of the methods described herein, the subject suffers from autism spectrum disorder, a disorder that shows one or more symptoms associated with autism spectrum disorder, social and communication disorder, or anxiety disorder.Examples of symptoms associated with autism spectrum disorder include but are not limited to persistent impairment of social communication and social interaction, social anxiety, and restricted repetitive behavior, interest, and activity.Other behaviors and characteristics observed in autism spectrum disorder also include aversion to physical contact, generalized anxiety, monotonous voice or inability to modulate voice volume, inability to establish equal relationships, lack of fun and sharing 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.

[0079] In some embodiments according to any of the methods described herein, the subject suffers from Prader-Willi syndrome.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 disability and learning difficulties, and many exhibit behavioral problems, including temper tantrums, stubbornness, misbehavior, and compulsive behaviors, including skin picking.Other symptoms commonly observed in individuals with Prader-Willi syndrome are persistent impairments in social communication and social interaction, anxiety and irritability, and sleep problems.In some embodiments,

[0080] In some embodiments according to any of the methods described herein, the subject suffers from one or more symptoms associated with Prader-Willi syndrome.Examples of symptoms associated with Prader-Willi syndrome include but are not limited to: persistent impairment of social communication and social interaction, anxiety and irritability, and sleep problems.In some embodiments according to any of the methods described herein, the subject suffers from anxiety associated with Prader-Willi syndrome.

[0081] In some embodiments according to any of the methods described herein, the subject suffers from a social and communication disorder. In some embodiments, the social and communication disorder is an impairment in communication skills and / or social interaction, a lack of eye contact, and / or an inability to form and / or maintain social relationships.

[0082] In some embodiments according to any of the methods described herein, the subject is suffering from anxiety associated with an autism spectrum disorder.

[0083] In some embodiments according to any of the methods described herein, the subject is suffering from pain, including, but not limited to, orofacial and craniofacial pain (e.g., headache), neck pain (e.g., occipital neuralgia), and pain in the upper extremities. In some embodiments, the pain is somatic pain. In some embodiments, the pain is superficial somatic pain. In some embodiments, the pain is deep somatic pain. In some embodiments, the pain is musculoskeletal pain. In some embodiments, the pain is visceral pain. In some embodiments, the pain is neuropathic pain. In some embodiments, the pain is cephalic or craniofacial pain. In some embodiments, the pain is in a body part other than the head and / or orofacial region. In some embodiments, the pain is chronic pain, such as a chronic pain described herein. In some embodiments, the pain is acute pain, such as an acute pain described herein. In some embodiments, the pain is a combination of one or more of the pains described herein. In some embodiments, the pain is sharp, shooting pain associated with movement. In some embodiments, the pain is neuropathic pain caused by nerve injury, such as nerve injury associated with surgery.

[0084] In some embodiments, the pain is head pain. In some embodiments, the pain is facial pain. In some embodiments, the pain is neck pain. In some embodiments, the pain is occipital neuralgia. In some embodiments, the pain is pain in the upper extremities. Examples of neck and upper extremity pain include, but are not limited to, nerve compression disorders (spinal stenosis), disc and vertebral disorders, diabetic neuropathy, cartilaginous tunnel syndrome, arthritic disorders, post-traumatic, occlusal facet disorders, post-herpetic neuralgia, and the like. In some embodiments, the pain is exacerbated by psychiatric disorders such as depression, anxiety, and stress. In some embodiments, the pain is induced or exacerbated by food (e.g., caffeine, chocolate, alcohol) or by drug overuse (e.g., opiates).

[0085] In some embodiments, the pain is trigeminal nerve-associated pain. Trigeminal nerve-associated pain can be selected from the group consisting of chronic, acute, and procedure-related pain, and combinations thereof. In some embodiments, the chronic pain is selected from the group consisting of trigeminal neuralgia, atypical facial pain, anesthesia painful, postherpetic neuralgia, head and neck cancer, migraine, and temporomandibular joint pain (TMJ). In some embodiments, the procedure-related pain is pain resulting from a dental, medical, surgical, or cosmetic procedure. In some embodiments, the acute pain is pain resulting from a laceration, burn, fracture, injury, headache, dental abscess, dental disease, bacterial infection, or sinus infection. Chronic, acute, or procedural pain associated with the trigeminal nervous system is experienced in many syndromes and disorders, including, but not limited to, trigeminal neuralgia, atypical facial pain, anesthesia painful, post-herpetic neuralgia, head and neck cancer, migraine, other types of headache, TMJ, injury to the face and / or head, dental injury or infection, general dental procedures, and facial surgery such as cosmetic plastic surgery.

[0086] Chronic pain in the face and head region can result from a variety of medical conditions, including, but not limited to, neuropathic pain, headache, TMJ, pain from cancer and / or cancer treatment. These pain syndromes are often not effectively treated with current medications or invasive interventions, and new methods for localized pain relief in the face and head region are needed.

[0087] In some embodiments, the methods described herein 13 The N-oxytocin peptide formulation or composition is directed to the trigeminal nervous system. 13 The N-oxytocin peptide formulation or composition is administered to a subject suffering from any of the diseases and conditions described herein.

[0088] In some embodiments of the methods described herein, the individual is a mammal. In some embodiments, the individual is a human. Oxytocin peptide preparation

[0089] In some embodiments,13 N-oxytocin peptide preparations, and 13 In some embodiments, there is provided a use of an N-oxytocin peptide formulation in determining the distribution and kinetics of oxytocin after application to craniofacial mucosa (such as application to the nose). 13 The N-oxytocin peptide formulation includes another agent. In some embodiments, the other agent is non-radiolabeled oxytocin. In some embodiments, the other agent affects the distribution, kinetics, and / or pharmacodynamics of oxytocin. In some embodiments, the other agent is a divalent metal cation (e.g., Mg 2+ , Ca 2+ , Zn 2+ , or Cu 2+ In some embodiments, the divalent metal cation is Mg 2+ In some embodiments, the other agent is vasopressin.

[0090] Oxytocin is known to treat head and craniofacial pain in humans and rats, and is particularly effective in treating chronic pain in rats where oxytocin receptors are overexpressed when administered intranasally.However, it has been observed that the analgesic effect of oxytocin in treating head pain, such as migraine, in human patients does not occur immediately after administration.Rather, there is an initial period of up to 2 hours before significant analgesia begins, and 4 hours are required for maximum analgesia, and patients continue to experience pain during this initial period.Various components of oxytocin formulations, such as magnesium and other divalent cations, can affect oxytocin therapy, including reducing pain intensity and modulating the onset and / or duration of analgesic effect.

[0091] In some embodiments, 13 N-oxytocin peptide preparations or compositions include: 13 In some embodiments, the liquid formulation comprises between about 0.01 mg / mL and about 16 mg / mL of oxytocin (including N-oxytocin peptide and non-radiolabeled oxytocin). 13N-oxytocin peptide preparations or compositions may contain between about 0.01 mg / mL and about 12 mg / mL, between about 0.05 mg / mL and about 16 mg / mL, between about 0.1 mg / mL and about 12 mg / mL, between about 0.1 mg / mL and about 8 mg / mL, between about 0.1 mg / mL and about 4 mg / mL, between about 0.1 mg / mL and about 2 mg / mL, between about 0.1 mg / mL and about 1.6 mg / mL, between about 0.1 mg / mL and about 1.2 mg / mL, between about 0.1 mg / mL and about 1 mg / mL, between about 0.1 mg / mL and about Between 0.8 mg / mL, between about 0.1 mg / mL and about 0.4 mg / mL, between about 0.1 mg / mL and about 0.3 mg / mL, between about 0.2 mg / mL and about 16 mg / mL, between about 0.2 mg / mL and about 12 mg / mL, between about 0.2 mg / mL and about 10 mg / mL, between about 0.2 mg / mL and about 8 mg / mL, between about 0.2 mg / mL and about 6 mg / mL, between about 0.2 mg / mL and about 4 mg / mL, between about 0.2 mg / mL and about 2 mg / mL, between about 0.2 mg / mL and about 1.6 mg / mL, between about 0 between about 0.2 mg / mL and about 1.2 mg / mL, between about 0.2 mg / mL and about 1 mg / mL, between about 0.2 mg / mL and about 0.8 mg / mL, between about 0.2 mg / mL and about 0.6 mg / mL, between about 0.2 mg / mL and about 0.4 mg / mL, between about 0.2 mg / mL and about 0.3 mg / mL, between about 0.3 mg / mL and about 16 mg / mL, between about 0.3 mg / mL and about 12 mg / mL, between about 0.3 mg / mL and about 10 mg / mL, between about 0.3 mg / mL and about 8 mg / mL, between about 0.3 mg / mL and about In some embodiments, the oxytocin concentration is between about 4 mg / mL, between about 0.3 mg / mL and about 3 mg / mL, between about 0.3 mg / mL and about 1 mg / mL, between about 0.3 mg / mL and about 0.5 mg / mL, between about 0.5 mg / mL and about 16 mg / mL, between about 0.5 mg / mL and about 10 mg / mL, between about 0.5 mg / mL and about 5 mg / mL, between about 0.5 mg / mL and about 1 mg / mL, between about 1 mg / mL and about 16 mg / mL, between about 1 mg / mL and about 10 mg / mL, or between about 1 mg / mL and about 5 mg / mL. 13N-oxytocin peptide formulations or compositions contain between about 0.1 mg / mL and about 2 mg / mL, between about 0.15 mg / mL and about 1.5 mg / mL, or between about 0.2 mg / mL and about 1.2 mg / mL of oxytocin. 13 The N-oxytocin peptide comprises the human oxytocin amino acid sequence consisting of Cys-Tyr-Ile-Gln-Asn-Cys-Pro-Leu-Gly (SEQ ID NO: 1). 13 The N-oxytocin peptide is a compound of formula (IV).

[0092] In some embodiments, 13 N-oxytocin peptide preparations or compositions include: 13 In some embodiments, the liquid formulation comprises about 5 IU / mL to about 8000 IU / mL of oxytocin (including N-oxytocin peptide and non-radiolabeled oxytocin). 13N-oxytocin peptide preparations or compositions may contain between about 500 IU / mL and about 6000 IU / mL, between about 25 IU / mL and about 8000 IU / mL, between about 50 IU / mL and about 6000 IU / mL, between about 50 IU / mL and about 4000 IU / mL, between about 50 IU / mL and about 2000 IU / mL, between about 50 IU / mL and about 1000 IU / mL, between about 50 IU / mL and about 800 IU / mL, between about 50 IU / mL and about 600 IU / mL, between about 50 IU / mL and about 500 IU / mL, and between about 50 IU / mL and about 400 IU / mL. between about 50IU / mL and about 200IU / mL, between about 50IU / mL and about 150IU / mL, between about 100IU / mL and about 8000IU / mL, between about 100IU / mL and about 6000IU / mL, between about 100IU / mL and about 5000IU / mL, between about 100IU / mL and about 4000IU / mL, between about 100IU / mL and about 3000IU / mL, between about 100IU / mL and about 2000IU / mL, between about 100IU / mL and about 1000IU / mL, between about 100IU / mL and about 800IU / mL, between about 100IU / mL and about Between 600 IU / mL, between about 100 IU / mL and about 500 IU / mL, between about 100 IU / mL and about 400 IU / mL, between about 100 IU / mL and about 300 IU / mL, between about 100 IU / mL and about 200 IU / mL, between about 100 IU / mL and about 150 IU / mL, between about 150 IU / mL and about 8000 IU / mL, between about 150 IU / mL and about 6000 IU / mL, between about 150 IU / mL and about 5000 IU / mL, between about 150 IU / mL and about 4000 IU / mL, between about 150 IU / mL and about 2000 IU / mL, In a preferred embodiment, the oxytocin concentration is between 50 IU / mL and about 1500 IU / mL, between about 150 IU / mL and about 500 IU / mL, between about 150 IU / mL and about 250 IU / mL, between about 250 IU / mL and about 8000 IU / mL, between about 250 IU / mL and about 5000 IU / mL, between about 250 IU / mL and about 2500 IU / mL, between about 250 IU / mL and about 500 IU / mL, between about 500 IU / mL and about 8000 IU / mL, between about 500 IU / mL and about 5000 IU / mL, or between about 500 IU / mL and about 2500 IU / mL. 13An N-oxytocin peptide formulation or composition comprises between about 50 IU / mL and about 1000 IU / mL, between about 75 IU / mL and about 750 IU / mL, or between about 100 IU / mL and about 600 IU / mL of oxytocin. 13 The N-oxytocin peptide comprises the human oxytocin amino acid sequence consisting of Cys-Tyr-Ile-Gln-Asn-Cys-Pro-Leu-Gly (SEQ ID NO: 1). 13 The N-oxytocin peptide is a compound of formula (IV).

[0093] Any divalent metal cation salt (e.g., a water-soluble divalent metal cation salt) can be used to prepare the divalent metal cation-containing salts of the present invention. 13 The N-oxytocin peptide formulation does not contain a divalent metal cation (e.g., Mg 2+ , Ca 2+ , Zn 2+ , or Cu 2+ ) can be supplied. 13 The divalent metal cation salt used in an N-oxytocin peptide formulation can be selected based on several factors, such as the amount of free divalent metal cation ions that can be delivered upon administration of the formulation, and for liquid formulations, the solubility of the divalent metal cation salt in the medium, the acidity / basicity of the counterion, and / or the dissociation constant of the salt. For example, in a liquid formulation, the divalent metal cation salt must be sufficiently soluble in the liquid medium to deliver the divalent metal cation ion concentration required to produce synergistic analgesia with the oxytocin peptide. Other factors, such as compatibility with other substances in the formulation and the ability of the counterion to perform other functions in the formulation, may also be taken into consideration when selecting a divalent metal cation salt. For example, magnesium citrate is sufficiently soluble in aqueous solution to provide the desired amount or concentration of divalent metal cations, citrate is pharmaceutically acceptable, citrate ions may act as a buffer, and magnesium citrate may also impart a pleasant flavor to the formulation. 13The divalent metal cation in the N-oxytocin peptide formulation may be provided using one or more divalent metal cation salts. 13 The divalent metal cation salt in the N-oxytocin peptide formulation contains 13 The divalent metal cations used in the preparation of N-oxytocin peptide formulations were the first to be identified. 13 In some cases, the divalent metal cation salt is generated in situ during the preparation of the N-oxytocin peptide formulation. For example, magnesium chloride may be used initially in the preparation of the formulation, and magnesium citrate may be generated in situ when citric acid is added to the formulation. In such cases, the divalent metal cation salt may be generated in situ during the preparation of the N-oxytocin peptide formulation. 13 The divalent metal cations in N-oxytocin peptide formulations are provided by both magnesium chloride and magnesium citrate.

[0094] Divalent metal cation-containing compounds as described herein 13 The divalent metal cation salts used in the N-oxytocin peptide formulations can be obtained from commercial sources or prepared according to methods known in the art. For example, magnesium citrate can be prepared according to the procedures described in Staszczuk P, et al. Physicochem Probl Mineral Proc 37: 149-158 (2003), U.S. Patent No. 1,936,364, and U.S. Patent No. 2,260,004.

[0095] In some embodiments, 13N-oxytocin peptide formulations or compositions may further comprise one or more pharmaceutically acceptable carriers (i.e., comprising a pharmaceutical composition) and, optionally, other ingredients, such as 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, formulations may include methyl-β-cyclodextrin (Me-β-CD), edetate disodium, arginine, sorbitol, NaCl, methylparaben sodium (MP), propylparaben sodium (PP), chlorobutanol (CB), benzyl alcohol, zinc chloride, ethyl alcohol, didecanoyl L-α-phosphatidylcholine (DDPC), polysorbate, lactose, citrate, tartrate, acetate, and / or phosphate.

[0096] Liquid carriers include, but are not limited to, water, saline, aqueous dextrose, and glycols, particularly for solutions (when isotonic). Carriers may 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, dried skim milk, glycerol, propylene glycol, water, ethanol, and the like. The compositions may be subjected to conventional pharmaceutical procedures, such as sterilization, and may contain conventional pharmaceutical additives, such as preservatives, stabilizers, reducing agents, antioxidants, chelating agents, wetting agents, emulsifiers, 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.

[0097] In some embodiments, 13 The N-oxytocin peptide formulation or composition may further comprise one or more mucosal delivery-enhancing agents selected from (A)-(K): (A) solubilizing agents, (B) charge modifiers, (C) pH adjusters, (D) degradative enzyme inhibitors, (E) mucolytic or mucus-clearing agents, (F) ciliostatic agents, (G) membrane permeability improvers, (H) modulators of epithelial junctional physiology, e.g., nitric oxide (NO) stimulators, chitosan, chitosan derivatives, (I) vasodilators, (J) selective transport enhancers, and (K). 13A stabilizing delivery vehicle, carrier, support, or complexing species to which an N-oxytocin peptide is effectively complexed, associated, housed, encapsulated, or bound to stabilize the active agent for enhanced mucosal delivery. The membrane permeability improver in group (G) may be (i) a surfactant, (ii) a bile salt, (iii) a phospholipid or fatty acid additive, mixed micelle, liposome, or carrier, (iv) an alcohol, (v) an enamine, (iv) an NO donor compound, (vii) a long-chain amphipathic molecule, (viii) a small hydrophobic permeability improver, (ix) a sodium or salicylic acid derivative, (x) a glycerol ester of acetoacetic acid, (xi) a cyclodextrin or a beta-cyclodextrin derivative, (xii) a medium-chain fatty acid, (xiii) a chelating agent, (xiv) an amino acid or a salt thereof, (xv) an N-acetyl amino acid or a salt thereof, (xvi) an enzyme degradative to a selected membrane component, (xvii) an inhibitor of fatty acid synthesis, (xviii) an inhibitor of cholesterol synthesis, or (xiv) a combination of any of the membrane permeability improvers of (i)-(xviii). 13 The N-oxytocin peptides can 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 can be used alone or together in a pharmaceutically acceptable formulation or delivery vehicle. 13 It may be mixed or otherwise combined with N-oxytocin peptides. 13 N-oxytocin peptide formulations or compositions, after delivery to a mucosal surface (e.g., in the nasal cavity) of a mammalian subject 13 The bioavailability of N-oxytocin peptides may be increased.

[0098] The list of carriers and excipients discussed herein is by no means complete, and those skilled in the art may select carriers and excipients from the GRAS (Generally Recognized As Safe) list of chemicals approved for pharmaceutical preparations, chemicals currently approved by the U.S. Food and Drug Administration for topical and parenteral formulations, and chemicals 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.)

[0099] In some embodiments, 13 The N-oxytocin peptide formulation or composition 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. 13 The N-oxytocin peptide formulation or composition further comprises chlorobutanol, acetic acid, and water.

[0100] In some embodiments, 13 The N-oxytocin peptide formulation or composition further comprises a chitosan-containing excipient (e.g., ChiSys®, http: / / www.archimedespharma.com / productArchiDevChiSys.html). In some embodiments, 13The N-oxytocin peptide formulation or composition further comprises about 1% 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 may be used, such as nanoparticles containing chitosan glutamate and a negatively charged polymer (e.g., pentasodium triphosphate). Thiolated chitosan (e.g., chitosan covalently modified with 2-iminothiolane), which has been used in microparticles containing insulin and reduced glutathione, may also be used in the preparations described herein. 13 N-oxytocin peptides may be useful as excipients in formulations or compositions.

[0101] In some embodiments, 13 N-oxytocin peptide formulations or compositions include 13 The N-oxytocin peptide formulation forms a gel in the nasal cavity, thus 13The formulations and methods described herein further include one or more gelling agents to enhance nasal absorption of the N-oxytocin peptide. Gelling systems useful in the formulations and methods described herein can include any known gelling system, such as a chemically reactive pectin-based gelling system (e.g., PecSys™, Archimedes Pharma) or a thermoresponsive polymer gelling system (e.g., Pluronic® F127, BASF). PecSys™ is a low-viscosity aqueous pectin-based solution delivered as a fine mist that gels upon contact of each droplet with calcium ions in the nasal mucosa. Other low-methoxy pectins can also be used, for example, at a concentration of about 1%. Pluronic® F127 contains 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. For example, Pluronic® F127 has been shown to gel in the human nasal cavity at approximately 18-20% wt / vol when used in a vitamin B12 gel supplement (EnerB, Nature's Bounty, NY) and in a gelling sumatriptan solution containing 18% wt / vol Pluronic® F127 and 0.3% wt / vol Carbopol (anionic bioadhesive polymer C934P). The monomer ratios and concentrations are determined by the predetermined 13 N-oxytocin formulations can be tailored to ensure gelation at 25-37°C, near the typical temperature of 34°C in the nasal cavity. If the gelation temperature is lower than 25°C, the formulation may gel at room temperature; if the gelation temperature is higher than 37°C, the formulation will not gel sufficiently upon contact with the nasal mucosa. In some embodiments, 13 The N-oxytocin peptide formulation or composition may further comprise a mucoadhesive agent, such as Carbopol. Addition of a mucoadhesive, for example, up to 0.5% Carbopol, can further reduce the gelling temperature.

[0102] In some embodiments, 13The N-oxytocin peptide formulation or composition further comprises a surfactant, such as a non-ionic surfactant (e.g., polysorbate 80), and one or more buffering agents, stabilizers, or tonicifiers. 13 The N-oxytocin peptide formulation or composition further comprises a propellant. The pH of the nasal spray solution is optionally between about pH 3.0 and 8.5, although the pH may be adjusted, if desired, to optimize delivery of the loaded macromolecular species (e.g., a therapeutic protein or peptide) in a substantially unionized state. The pharmaceutical solvent employed may also be a weakly acidic aqueous buffer (pH 3-6). Suitable buffers for use in such compositions are described above or otherwise known in the art. Other components, including preservatives, surfactants, dispersing agents, or gases, may be added to promote or maintain chemical stability. Suitable preservatives include, but are not limited to, phenol, methylparaben, parabens, m-cresol, thiomersal, benzalkonium chloride, and the like. Suitable surfactants include, but are not limited to, oleic acid, sorbitan trioleate, polysorbates, lecithin, phosphatidyl choline, and various long-chain diglycerides and phospholipids. Suitable dispersing agents include, but are not limited to, ethylenediaminetetraacetic acid (EDTA). Suitable gases include, but are not limited to, nitrogen, helium, chlorofluorocarbons (CFCs), hydrofluorocarbons (HFCs), carbon dioxide, air, and the like. Suitable stabilizers and isotonicity agents include sugars and other polyols, amino acids, and organic and inorganic salts. In some embodiments, 13 The N-oxytocin peptide formulation or composition further comprises a citrate, a succinate, or a pyrophosphate salt.

[0103] 13To further enhance mucosal delivery of N-oxytocin peptide, enzyme inhibitors, particularly protease inhibitors, may be further included in the formulation.Protease inhibitors may include, but are not limited to, antipain, alphamenin 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, holoxymitin, 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.

[0104] 13 Absorption enhancers may be included in the formulation to enhance delivery and / or absorption of N-oxytocin peptides into or through mucosal surfaces. Such enhancers may enhance the composition's release or solubility (e.g., from the formulation delivery vehicle), diffusion rate, penetration 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). Thus, enhanced mucosal delivery may include, for example: 13 This may occur through any of a variety of mechanisms, including by increasing the diffusion, transport, persistence, or stability of N-oxytocin peptides, increasing membrane fluidity, modulating the availability or action of calcium and other ions that regulate intracellular or paracellular permeability, solubilizing mucosal membrane components (e.g., lipids), altering non-protein and protein sulfhydryl levels in mucosal tissues, increasing water flux across mucosal surfaces, modulating epithelial junctional physiology, reducing the viscosity of mucus overlying the mucosal epithelium, slowing the rate of mucociliary clearance, and other mechanisms.

[0105] Compounds that enhance mucosal absorption can include, but are not limited to, surfactants, bile salts, dihydrofusidate salts, bioadhesives / mucoadhesives, phospholipid additives, mixed micelles, liposomes, or carriers, alcohols, enamines, cationic polymers, NO donor compounds, long-chain amphiphilic molecules, small hydrophobic permeability 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 that specifically target selected membrane components, inhibitors of fatty acid synthesis, and inhibitors of cholesterol synthesis.

[0106] All peptides described and / or contemplated herein can be prepared by chemical synthesis using either automated or manual solid phase synthesis techniques generally known in the art. The peptides may also be prepared using recombinant molecular techniques known in the art. Oxytocin administration

[0107] 13The N-oxytocin peptide formulation or composition may be adapted for craniofacial mucosal administration (e.g., nasal, buccal, sublingual, or ophthalmic administration). In some embodiments, the composition may be adapted for use with a device for mucosal delivery. In some embodiments, the composition may be adapted for buccal and / or sublingual mucosal delivery and may be intended for use with a device for buccal and / or sublingual mucosal administration, such as a unit-dose container, a pump spray, a dropper, a squeeze bottle, an airless and preservative-free spray, a nebulizer, a metered-dose inhaler, or a pressurized metered-dose inhaler. In some embodiments, the composition may be adapted for ophthalmic delivery and may be intended for use with a device for conjunctival administration, such as a dropper or squeeze bottle. In some embodiments, the composition may be adapted for intranasal administration and may be intended for use with a device for intranasal administration, such as a dropper, a pump spray, a squeeze bottle, an airless and preservative-free spray, or a nasal pump device, e.g., a nasal pump device comprising a reservoir bottle attached to an aerosolization device.

[0108] 13In embodiments in which an N-oxytocin peptide formulation or composition is administered intranasally, the composition can be prepared as a liquid aerosol formulation, optionally combined with a dispersing agent and / or a physiologically acceptable diluent. Alternatively, dry powder aerosol formulations are contemplated and may contain a finely divided solid form of the subject compound and a dispersing agent that allows for the immediate 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 mucous membranes of the nasal passages or lungs. The term "aerosol particles" is used herein to describe suitable liquid or solid particles of a size small enough to distribute to the targeted mucous membranes or alveolar membranes in the nose or lungs. Other considerations include the design of the delivery device, additional components in the formulation, and particle characteristics. These aspects of nasal or pulmonary drug administration are well known in the art, and the manipulation of formulations, aerosolization means, and delivery device design are within the skill of those in the art.

[0109] 13 In embodiments in which the N-oxytocin peptide formulation or composition is administered intranasally, administration is accomplished by an intranasal delivery device. 13 The device may be any suitable device for intranasal administration of an N-oxytocin peptide formulation. In some embodiments, the device comprises: 13 The device is suitable for delivering N-oxytocin peptide to specific regions within the nasal cavity. In some embodiments, the device comprises: 13 The device is suitable for delivering N-oxytocin peptide to the lower two-thirds of the nasal cavity. 13 The device is suitable for delivering N-oxytocin peptide to the upper third of the nasal cavity. 13 Suitable for delivering N-oxytocin peptide throughout the nasal passages.

[0110] In some embodiments, the intranasal delivery device 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 a specified volume (e.g., about 50 to about 150 μL, preferably about 50 μL or about 100 μL) with a specified droplet size distribution. In some embodiments, the nasal pump device includes a reservoir bottle attached to an aerosolization device, such as the Equadel pump marketed by Aptar Pharma. In some embodiments, the nasal administration device functions regardless of the pressure applied to the pump once a threshold is reached. For administration in larger mammals, the nasal pump device may include a reservoir bottle attached to a pump actuator that meters a larger volume (e.g., about 100 μL to about 600 μL, or more).

[0111] In some embodiments, the intranasal delivery device is designed to deliver multiple doses of a drug formulation. For example, a nasal pump device may include a reservoir bottle attached to a pump actuator, which holds multiple doses of the liquid formulation, and the pump actuator may meter a specified volume that is a fraction of the liquid formulation held in the reservoir bottle. In some embodiments, the pump actuator may meter approximately 50 μL of the liquid formulation per spray. The nasal pump device may include a filter to prevent backflow to reduce (e.g., bacterial) contaminants from entering the reservoir bottle. In some embodiments, the nasal pump device includes a metal-free passageway (e.g., a plastic passageway) for delivering the liquid formulation. In some embodiments, the pump device uses a plastic material that is stable to gamma radiation (used to sterilize nasal devices). In some embodiments, the intranasal delivery device includes a multi-dose pump that includes a microbial filter and an automatic sealing mechanism within the pump actuator, such as the spray device described in U.S. Pat. No. 5,988,449.

[0112] In some embodiments, the intranasal delivery device is a breath-activated nasal delivery device, such as those described in U.S. Patent Nos. 7,784,460 and 7,854,227. Such devices may improve delivery to target sites deep within the nasal cavity. In some embodiments, a standard metered-dose spray device is incorporated into a housing that allows the patient to actuate the device by blowing into the mouthpiece. In some embodiments, the device consists of a conical, sealing nosepiece and mouthpiece, incorporating a traditional mechanical spray pump (e.g., the Equadel pump marketed by Aptar Pharma), a chargeable spring, and a breath-activated mechanism. This mechanism can be used for single-dose or multi-dose delivery. An example of such a liquid delivery device is the OptiMist™ device marketed by OptiNose. During use, the nasal portion of the device is inserted into the nostrils and the mouth portion is blown into. This closes the soft palate, transferring pressure to the nostrils and opening the passages, providing airflow behind the nasal septum and allowing air to exit through the other nostril (two-way flow). The device is breath-activated, preventing small particles from entering the lungs. By adjusting the flow rate and particle size, specific nasal areas can be targeted.

[0113] In some embodiments, the intranasal delivery device comprises: 13 The intranasal delivery device is a unit-dose metered-dispense spray device suitable for single administration of an N-oxytocin peptide formulation or composition. In some embodiments, the intranasal delivery device comprises: 13 A multi-dose metered-delivery spray pump device suitable for repeated administration of N-oxytocin peptide.

[0114] Droplet size, plume volume, and flow rate can be adjusted to target specific nasal regions. To target the olfactory and / or respiratory epithelium, liquid sprays may achieve droplet sizes between 5 and 50 microns. Larger droplets primarily travel down the nasopharynx and are swallowed, while smaller droplets are directed to the lung tissue. The mass median equivalent aerodynamic diameter (MMAD) is used to specify droplet size. The pH of nasal sprays is optimized to deliver the loaded peptides in a largely unionized state. The nose generally tolerates solutions with a pH of approximately 3 to 8. The nasal mucosa can generally absorb a volume of approximately 100 μL before saturation occurs and the liquid begins to drip from the nose. Therefore, plume volumes may be up to and including 100 μL. For use in large mammals, plume volumes may be up to and including 150 μL or larger (e.g., 600 μL or larger). For use in infants and children, or for veterinary use in smaller animals (e.g., rodents, cats), smaller plume volumes (5-50 μL) may be used.

[0115] In some embodiments, the intranasal delivery device is ergonomically designed to promote patient compliance, such as a pump device with a side-actuation trigger mechanism. In some embodiments, the intranasal delivery device includes a metered-dose spray pump that acts as a sealed system, preventing air from entering the pump device and thus preventing contamination with airborne microorganisms. In some embodiments, the intranasal delivery device includes a metered-dose spray pump that works in conjunction with a filter. Exhaled air is drawn through a filter incorporated within the pump, and airborne microorganisms are kept outside 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 treatment monitoring.

[0116] In some embodiments, 13 N-oxytocin peptide formulations or compositions include 13 Contains N-oxytocin peptide, 13 The N-oxytocin peptide is contained in any one of the intranasal delivery devices described herein; 13 N-oxytocin peptide preparation or composition 13 The concentration of oxytocin (including N-oxytocin and non-radiolabeled oxytocin) is within any of the concentration ranges described herein, as if each and every combination of device and concentration were individually described. In some embodiments, 13 N-oxytocin peptide formulations or compositions include 13 In some embodiments, the compound comprises an N-oxytocin peptide and non-radiolabeled oxytocin. 13 N-oxytocin peptide preparations or compositions 13 The amount of N-oxytocin peptide is in the range of about 0.05 μg / mL to about 5 μg / mL (e.g., about any of 0.05, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, or 5 μg / mL; including any ranges therebetween).

[0117] In some embodiments, 13 The N-oxytocin peptide is administered simultaneously or sequentially with another agent. In some embodiments, the other agent is non-radiolabeled oxytocin. In some embodiments, the other agent affects the distribution, kinetics, and / or pharmacodynamics of oxytocin. In some embodiments, the other agent is a divalent metal cation (e.g., Mg 2+ , Ca 2+ , Zn 2+ , or Cu 2+ In some embodiments, the divalent metal cation is Mg 2+ In some embodiments, the other agent is vasopressin. 13 The N-oxytocin peptide is co-administered with the other agent in the same unit dose. 13The N-oxytocin peptide is co-administered with the other agent as a separate unit dose or formulation. 13 The N-oxytocin peptide and the other agent are administered separately. In some embodiments, the other agent is administered to the subject in a first administration, and then 13 An N-oxytocin peptide is administered to the subject in a second administration. 13 The N-oxytocin peptide is administered between about 10 minutes and about 2 hours after administration of the other agent. 13 The N-oxytocin peptide is administered between about 10 minutes and about 2 hours, between about 10 minutes and about 1 hour, between about 10 minutes and about 30 minutes, between about 20 minutes and about 2 hours, between about 20 minutes and about 1 hour, between about 30 minutes and about 2 hours, or between about 30 minutes and about 1 hour after administration of the other agent. 13 The N-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 other agent. 13 The N-oxytocin peptide is administered about 10 minutes, about 15 minutes, about 20 minutes, or about 30 minutes after the administration of the other agent. 13 The N-oxytocin peptide is administered to the subject first, and then the other agent is administered to the subject. In some embodiments, the subject is a human.

[0118] In some embodiments, 13 The N-oxytocin peptide and the other agent may be administered to a subject in need thereof by the same route or by different routes. 13 The N-oxytocin peptide is administered by craniofacial mucosal administration (e.g., nasal, buccal, sublingual, or ophthalmic administration). 13 The N-oxytocin peptide and the other agent are both administered intranasally in the same formulation. 13 N-oxytocin peptides are administered via the craniofacial mucosa, while other agents are administered systemically, eg, intravenously, intramuscularly, orally, subcutaneously, or intrathecally.

[0119] In some embodiments, 13 The N-oxytocin peptide is administered by intranasal administration. 13 N-oxytocin peptides and other drugs are administered by intranasal administration. 13 N-oxytocin peptides and / or other agents can be administered to the mucosal tissues in the nasal cavity using a suitable intranasal delivery device, such as the nasal delivery device described herein. Suitable areas in the nasal cavity include, but are not limited to, the lower two-thirds, the upper one-third, or the entire nasal passage. In some embodiments, 13 The N-oxytocin peptide and / or other agent is administered to the upper third of the nasal cavity. 13 The N-oxytocin peptide and / or other agent is administered to the lower two-thirds of the nasal cavity. 13 The N-oxytocin peptide and / or other agent is specifically delivered to reach both the lower two-thirds and the upper one-third of the nasal cavity.

[0120] In some embodiments, 13 The N-oxytocin peptide comprises the human oxytocin amino acid sequence consisting of Cys-Tyr-Ile-Gln-Asn-Cys-Pro-Leu-Gly (SEQ ID NO: 1). 13 The N-oxytocin peptide is a compound of formula (IV). 13 An effective dose of an N-oxytocin peptide formulation or composition ranges from about 0.5 μg to about 2000 μg of oxytocin ( 13 In some embodiments, the oxytocin comprises: 13 The effective dose of oxytocin in the N-oxytocin peptide formulation or composition is about 0.5 μg to about 1000 μg, about 1 μg to about 1000 μg, or about 1 μg to about 2000 μg. 13The effective dose of oxytocin in the N-oxytocin peptide preparation or composition 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 4 00μg, about 16μg to about 200μg, about 16μg to about 160μg, about 16μg to about 120μg, about 16μg to about 80μg, about 20μg to about 1 000μg, about 20μg to about 800μg, about 20μg to about 500μg, about 20μg to about 200μg, about 20μg to about 100μg, about 30μg to about In some embodiments, the amount is 1000 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. 13 The effective dose of oxytocin in the N-oxytocin peptide formulation or composition 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. 13 The effective dose of oxytocin in the N-oxytocin peptide formulation or composition 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. 13 In an effective dose of an N-oxytocin peptide formulation or composition 13 The amount of N-oxytocin peptide is approximately 1 × 10 -4 μg ~ approx. 1×10 -2 μg (e.g., approximately 1 × 10 -4, 5×10 -3 , 1×10 -3 , 5×10 -2 , or 1 × 10 -2 μg; and any ranges therebetween).

[0121] In some embodiments, 13 Oxytocin ( 13 An effective dose of oxytocin (including N-oxytocin and non-radiolabeled oxytocin) is from about 0.25 IU to about 1000 IU. 13 An effective dose of oxytocin in an N-oxytocin peptide formulation or composition 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. 13 The effective dose of oxytocin in the N-oxytocin peptide preparation or composition is about 2 IU to about 500 IU, about 4 IU to about 500 IU, about 4 IU to about 400 IU, about 4 IU to about 250 IU, about 4 IU to about 200 IU, about 4 IU to about 150 IU, about 4 IU to about 100 IU, about 4 IU to about 50 IU, about 4 IU to about 40 IU, about 4 IU to about 25 IU, about 5 IU to about 500 IU, about 5 IU to about 250 IU, about 5 IU to about 100 IU, about 5 IU to about 50 IU, about 8 IU to about 500 IU, about 8 IU to about 400 IU, about 8 IU to about 250 IU, about 8 IU to about 20 In some embodiments, the dose is 0 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. 13An effective dose of oxytocin in an N-oxytocin peptide formulation or composition 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 some embodiments, 13 An effective dose of oxytocin in an N-oxytocin peptide preparation or composition 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. kit

[0122] Provided herein are kits for carrying out any of the methods described herein. The kits include: 13 In some embodiments, the kit comprises a peptide according to formula (IV), such as a peptide of formula (IV), for use in administering N-oxytocin to an individual (e.g., craniofacial mucosal administration, e.g., intranasal administration). 13 In some embodiments, the kit comprises: 13 Peptides according to formula (I), (Ia), (Ib), (II), (IIa), and / or (IIb) used in the synthesis of N-oxytocin peptides 13 In some embodiments, the kit comprises a N-oxytocin peptide precursor. 13 The kit includes reagents used in the synthesis of N-oxytocin peptide, including DECP, PMP, DMSO, THF, HCl, and / or dioxane. 13The kit includes reagents and / or articles of manufacture, including an SPE column (e.g., C18 sep-pak) and / or an ion-pairing reagent, for purifying N-oxytocin peptide. In some embodiments, the kit includes a device for craniofacial mucosal administration (e.g., intranasal administration) in suitable packaging. The kit may further include any of the oxytocin peptide formulation components described herein, including a protease inhibitor and / or at least one absorption enhancer. Other kits may further include instructions for use that provide information to a user and / or health care provider regarding the practice of any one of the methods described herein.

[0123] A method for administering a compound as described herein to a facial mucosa, the method comprising ... 13 Kits are also provided that include an N-oxytocin peptide formulation and suitable packaging materials. 13 It may further comprise instructions for administration of the N-oxytocin peptide formulation.

[0124] The instructions for using the kit to practice the invention generally describe how to use the contents of the kit to practice the methods of the invention. The instructions provided in the kits of the invention will typically be written instructions on a label or package insert (e.g., a paper sheet included with the kit), although machine-readable instructions (e.g., instructions contained on a magnetic or optical storage disk) are also acceptable. Illustrative Embodiments

[0125] Embodiment 1. Comprising the amino acid sequence of SEQ ID NO: 1 13 N-labeled oxytocin peptide.

[0126] Embodiment 2. One composition of oxytocin peptide 14 N atom 13 of embodiment 1, wherein the N radionuclide is replaced by 13 N-labeled oxytocin peptide.

[0127] Embodiment 3. One residue of the oxytocin peptide is: 13 3. The method of claim 1 or 2, wherein the compound is modified to include a moiety containing a N radionuclide. 13 N-labeled oxytocin peptide.

[0128] Embodiment 4. An oxytocin peptide wherein one residue is: 13 3. The compound of claim 3, wherein the compound is modified to include NH. 13 N-labeled oxytocin peptide.

[0129] Embodiment 5. Single 13 5. Any one of embodiments 1 to 4, comprising a N radionuclide. 13 N-labeled oxytocin peptide.

[0130] Embodiment 6. a) a glutamine residue at position 4 (SEQ ID NO: 2), b) an asparagine residue at position 5 (SEQ ID NO: 3), or c) a glycine residue at position 9 (SEQ ID NO: 4). 13 6. Any one of embodiments 1 to 5, comprising a N radionuclide. 13 N-labeled oxytocin peptide.

[0131] Embodiment 7. The compound of embodiment 1, which is a compound of formula (IV). 13 N-labeled oxytocin peptide.

[0132] Embodiment 8. Comprising the amino acid sequence of SEQ ID NO: 1 13 1. A method for preparing an N-labeled oxytocin peptide, comprising: a) reacting a compound of formula (Ib) with diethyl cyanophosphonate (DECP) to provide a compound of formula (IIb); and b) treating the compound of formula (IIb) with a gas. 13 NH to provide a compound of formula (IIIb); and c) deprotecting the compound of formula (IIIb); 13 providing an N-labeled oxytocin peptide, 13 The method, wherein the N-labeled oxytocin peptide is a compound of formula (IV):

[0133] Embodiment 9 The method of embodiment 8, wherein the reaction of the compound of Formula (Ib) with DECP is carried out in the presence of dimethyl sulfoxide (DMSO), tetrahydrofuran (THF), and pentamethylpiperidine (PMP).

[0134] Embodiment 10. The method of embodiment 8 or 9, wherein the amount of DECP is between about 0.7 equivalents and about 1.1 equivalents.

[0135] Embodiment 11 The method of embodiment 9 or 10, wherein the ratio of DMSO to THF is between about 1:7 and about 1:11.

[0136] Embodiment 12 The method of any one of embodiments 8 to 11, wherein deprotecting comprises reacting the compound of Formula (IIIb) with HCl / dioxane.

[0137] Embodiment 13. The method provided in step c) 13 13. The method of any one of embodiments 8 to 12, further comprising purifying the N-labeled oxytocin peptide to remove reagents, organic solvents, and precursors.

[0138] Embodiment 14 The method of embodiment 13, wherein purifying comprises purifying by solid phase extraction (SPE).

[0139] Embodiment 15. Purification by SPE comprises: a) 13 The N-labeled oxytocin peptide is applied to a first hydrophobic SPE column, 13 allowing N-oxytocin and precursors to be retained on a first hydrophobic SPE column; and b) applying a solution comprising an aqueous ion-pairing reagent to the first hydrophobic SPE column; 13 allowing N-oxytocin to elute in a first eluate; and c) applying the first eluate to a second hydrophobic SPE column to obtain a second hydrophobic SPE column. 13 allowing N-oxytocin to be retained on a second hydrophobic SPE column; and d) 13 and eluting N-oxytocin in a second eluent.

[0140] Embodiment 16 The method of embodiment 15, wherein the first and / or second hydrophobic SPE column comprises a silica-based bonded phase having strong hydrophobic properties.

[0141] Embodiment 17. The method of embodiment 15 or 16, wherein the solution containing the aqueous ion-pairing reagent comprises between about 15% and about 25% acetonitrile.

[0142] Embodiment 18. Prepared by a process comprising the method of any one of embodiments 8 to 17. 13 N-labeled oxytocin peptide.

[0143] Embodiment 19. A method of determining the distribution of exogenously administered oxytocin in an individual, comprising: a) administering to the individual a dose of any one of embodiments 1 to 8 and 18. 13 administration of N-labeled oxytocin peptide; and b) 13 allowing the N-labeled oxytocin peptide to accumulate at a tissue or cell site and be imaged; and c) imaging the cell or tissue by a non-invasive imaging technique.

[0144] Embodiment 20. A method for determining the distribution of oxytocin receptors in an individual, comprising: a) administering to the individual a method according to any one of embodiments 1 to 8 and 18. 13 administration of N-labeled oxytocin peptide; and b) 13 allowing the N-labeled oxytocin peptide to bind to an oxytocin receptor; and c) in the individual. 13 and imaging the N-labeled oxytocin peptide by a non-invasive imaging technique.

[0145] Embodiment 21. A method of determining the kinetics of exogenously administered oxytocin in an individual, comprising: a) administering to the individual a dose of any one of embodiments 1 to 8 and 18. 13 and b) administering N-labeled oxytocin peptide to the individual. 13and imaging the N-labeled oxytocin peptide over a period of time by a non-invasive imaging technique.

[0146] Embodiment 22. The method of any one of embodiments 19 to 21, wherein the non-invasive imaging technique comprises positron emission tomography imaging.

[0147] Embodiment 23. The method of embodiment 22, wherein the non-invasive imaging technique comprises positron emission tomography with computed tomography imaging or positron emission tomography with magnetic resonance imaging.

[0148] Embodiment 24. 13 24. The method of any one of embodiments 19 to 23, wherein the N-labeled oxytocin peptide is administered by craniofacial mucosal administration.

[0149] Embodiment 25. 13 25. The method of embodiment 24, wherein the N-labeled oxytocin peptide is administered intranasally.

[0150] Embodiment 26. 13 24. The method of any one of embodiments 19 to 23, wherein the N-labeled oxytocin peptide is administered intravenously, intraarterially, intraperitoneally, intravesically, subcutaneously, intrathecally, intrapulmonary, intramuscularly, intratracheally, ophthalmically, transdermally, or via suppository.

[0151] Embodiment 27. 13 27. The method of embodiment 26, wherein the N-labeled oxytocin peptide is administered intravenously.

[0152] Embodiment 28. Any one of embodiments 1 to 7 and 18 13 Kit containing N-labeled oxytocin peptide. [Example]

[0153] Those skilled in the art will recognize that several embodiments are possible within the scope and spirit of the present invention. The present invention will now be described in more detail by reference to the following non-limiting examples. The following examples further illustrate the present invention but, of course, should not be construed as in any way limiting its scope. Example 1 Distribution of oxytocin in rats after intravenous administration.

[0154] Anesthetized rats were scanned using a microPET scanner (nanoscan microPET / 1T MRI, Mediso Ltd, Budapest, Hungary). Body temperature was maintained, and circulation and respiration were monitored. First, structural T2-weighted MRI scans were obtained for attenuation correction. Subsequently, rats were injected with 13N-oxytocin radiotracer via the tail vein. A list-mode protocol with an acquisition time of 60 min was used. The list-mode data from the emission scans were divided into dynamic series frames. The data were reconstructed for each time frame using iterative reconstruction. Appropriate correction factors were applied to generate quantitative dynamic images (matrix size (x, y, z) 93, 93, 120; voxel size 0.4 mm; 3D Tera-Tomo reconstruction with Monte Carlo-based estimation, attenuation, and scatter correction). The summed PET images from each 60-minute scan were co-registered to a standard Sprague-Dawley rat MRI brain atlas using PMOD software (version 3.7, PMOD Technologies Ltd, Zurich, Switzerland), a commercially available software package for quantifying biomedical images, to acquire data in the regions of interest. Results are presented as the dimensionless standard uptake value (SUV): [tissue activity concentration (MBq / g) × body weight (g) / injected dose (MBq)].

[0155] In one study, female Sprague Dawley rats were housed two per cage in a 12-hour light / 12-hour dark cycle, with an average temperature of 21°C and humidity of 55%, and had free access to food and water. Animal experiments were conducted under humane conditions with ethical approval and in accordance with guidelines established by the Danish Animal Experiments Inspectorate (AEI).

[0156] Female Sprague Dawley rats (weight: 358 g) were anesthetized in a chamber filled with 2% isoflurane and maintained via an isoflurane delivery mask (1.5-2%) during dynamic PET acquisition. Body temperature, heart rate, and respiratory rate were monitored throughout the study. Anesthetized animals were scanned using a microPET scanner (nanoScan® microPET / 1T MRI System, Mediso Ltd, Budapest, Hungary) (MRI image resolution 100 μm) by placing the animal prone in the microPET aperture. The head was fixed in a holder to minimize head movement. First, structural T2-weighted MRI scans were obtained for attenuation correction. Subsequently, the rats received 13.3 MBq of MRI. 13 N-oxytocin radiotracer was injected via the tail vein and PET scanning was initiated. Data were acquired in PET list mode and analyzed using a 3D iterative algorithm (Tera-Tomo 3D, full detector model and normal regularization (Mediso, Budapest, Hungary)) with four iterations and six subsets, resulting in a 0.4 × 0.4 × 0.4 mm (0.064 mm) area. 3 The images were reconstructed into 20 frames (30 min (8 × 15 s, 8 × 60 s, 4 × 5 min)) with a voxel size of 15 s. A delayed matching window was used to correct the data for dead time and attenuation as well as chance events. Images were corrected for attenuation and scatter using an 18-min long MR GRE EXT sequence (single excitation, TR 2.0 ms, TE 2.1 ms, flip angle 25°, 0.5 mm slice thickness, horizontal orientation). Observation of the acquired data showed that 13No uptake of N-oxytocin into the brain was observed (Fig. 2). Example 2 Distribution of oxytocin after intranasal administration in rats.

[0157] In rats 13 The distribution of N-oxytocin after nasal administration was studied in rats using any of the formulations described herein. 13 N-oxytocin-containing aqueous preparation was administered intranasally to rats. 13 The distribution of N-oxytocin is determined by scanning as described above.

[0158] In one study, two rats (weighing 257 g and 260 g) 13 The distribution of N-oxytocin after intranasal administration was investigated. For each rat, approximately 100 μl of 13 N-oxytocin solution (dissolved in 10% ethanol / 90% saline) (approximately 1-2 MBq) was pipetted into each nostril, and the rat was immediately transferred to the PET scanning bed. After that, a 60-minute PET scan was initiated according to the method described in Example 1, followed by magnetic resonance (MR) scanning. As shown in Figure 3, the tracer 13 N-oxytocin was distributed primarily to the gallbladder, kidney, and intestine, with no appreciable brain uptake. Example 3 Distribution and kinetics of oxytocin after intranasal administration in healthy human subjects

[0159] In healthy human subjects 13 The distribution and kinetics of N-oxytocin following intranasal administration are studied. Healthy volunteers are selected based on inclusion / exclusion criteria, medical history and physical examination, laboratory tests, and other routine procedures. 13 N-oxytocin is delivered intranasally to subjects by a metered dose nebulizer. For example, each nasal spray application delivers a dose of 0.01 μg / kg in 0.1 ml of saline. 13 N-oxytocin is administered to the subjects. Control subjects receive only 0.1 ml of saline with each nasal spray application.13 Any N-oxytocin formulation may be tested. 13 N-oxytocin was imaged over time by PET. 13 The distribution and dynamics of N-oxytocin are revealed. In this example, a structural CT or MRI is first performed to accurately confirm the anatomy. 13 After administering N-oxytocin to the subject, nasal rinsing was performed to allow for accurate timing of the oxytocin pulse. A 60-minute PET scan was initiated immediately after the healthy volunteer, migraine patient, or neuropsychiatric patient was properly positioned in the PET scanning device (Siemens Biograph 64 Truepoint PET). PET data were reconstructed using TrueX 3D OSEM (3 iterations, 21 subsets), a 256 × 256 × 10 matrix, and a 2 mm Gaussian filter, with a time window structure of 5 × 60, 3 × 300, and 4 × 600 seconds (12 frames total, 60 minutes). A preprocessing step was performed using PMOD 3.7 (PMOD Technologies Ltd, Zurich, Switzerland), a commercially available software package for quantifying biomedical images. Dynamic PET acquisitions were visually inspected and motion artifacts corrected. A whole-body radiation scan was performed on all six participants. Take safety tests, blood, urine, vital signs, and electrocardiogram recordings before and between 120 and 150 minutes after injection.

[0160] For whole-body images, ROIs are placed on major organs (brain, lungs, mediastinum, liver, spleen, gallbladder, kidneys, bladder, and ureters). The activity in each source organ and the remainder is divided by the injected activity to determine uptake as a percentage of injected activity. The residence time in each source organ is calculated by fitting an exponential curve using OLINDA 1.0 / EXM software (Vanderbilt University) from the percent injected activity and time course information for each source organ. Using an adult male phantom, dose measurements are calculated by entering the mean residence time for each source organ into OLINDA 1.0 / EXM software. Example 4 Distribution, kinetics, and pharmacodynamics of oxytocin after intranasal administration in healthy human volunteers, neuropsychiatric patients, or patients with chronic migraine

[0161] Human subjects (healthy volunteers, patients suffering from migraine with or without aura, or neuropsychiatric patients) will be enrolled. Study eligibility will be determined by telephone interview and upon their first visit to the research center. When potential participants first visit the research center, a medical history will be taken and a physical examination will be performed. Particularly, patients with neuropsychiatric disorders (e.g., autism spectrum disorder or anxiety disorder), chronic headache with or without aura, and other conditions will be considered. A diagnosis of episodic migraine or frequent episodic migraine has been established by one skilled in the art. If participants meet the inclusion criteria and do not meet the exclusion criteria, written informed consent is obtained. Demographic and medical data are recorded.

[0162] The pain assessment tool used in migraine patients is the VRS-4. This tool is self-reported and requires subjects to grade their pain level on a 4-category scale (severe, moderate, mild, none). Subjects enter their pain score using a secure, portable ePRO device. Subjects are trained on administering the VRS-4 using the portable device during the screening visit.

[0163] For neuropsychiatric patients, appropriate research assessments will be used, such as the Autism Diagnostic Observation Scale, the Clinical Global Impression-Improvement (CGI-I) assessment, and / or the Hamilton Anxiety Rating Scale.

[0164] The effectiveness of the oxytocin preparation is assessed by appropriate measurement tools by comparing the effect after administration compared to before treatment, as well as the time to onset of effect and duration of effect.

[0165] First, subjects undergo structural MRI or CT scans, then receive regular intranasal administration of an oxytocin peptide preparation. 13 Contains unlabeled oxytocin in addition to N-oxytocin. 13 After administering the N-oxytocin peptide formulation, the subject is imaged by PET. 13 The kinetics and distribution of N-oxytocin are determined. PET images are analyzed to determine dosimetry and pharmacokinetics and the relationship of these measures to pharmacodynamics (with respect to efficacy). Scanning and analysis are performed as described above. Example 5 13 Method for synthesizing N-oxytocin

[0166] In the following examples, the compound of formula (Ib) was used as the starting reagent to prepare a carboxy-terminal 13 of formula (IV) containing NH 13 A method for synthesizing the N-oxytocin compound is described. The compound of formula (Ib) was incubated in a reaction vial with 0.9 equivalents of DECP in the presence of dimethyl sulfoxide (DMSO)-tetrahydrofuran (THF) (1:9 ratio, respectively) containing pentamethylpiperidine (PMP) at room temperature to produce the compound of formula (IIb). In a cyclotron, a liquid target ([ 16 O(p,α) 13 N) by proton irradiation 13 NH3 was generated and transferred through Teflon tubing with a helium flow to a glass vial containing neat NaOH. 13 NH3 was manually transferred by positive pressure generated by a syringe into the reaction vial containing the compound of formula (IIb) at room temperature to give the N-Boc protected compound of formula (IIIb). The N-Boc protected compound of formula (IIIb) was deprotected using 4 M HCl / dioxane at 50 °C for 2.5 minutes to give the compound of formula (IV). 13 A crude preparation of N-oxytocin compound was obtained.

[0167] of formula (IV) 13The crude preparation of N-oxytocin compound was purified using a multi-step solid phase extraction (SPE) protocol to remove reagents, organic solvents, and precursors. In the first step, the compound of formula (IV) 13 A diluted crude preparation of N-oxytocin compound was applied to a C18 Sep-pak cartridge, allowing the organic solvent and reagents to flow through in the eluent, while 13 N-oxytocin and precursors were retained in the cartridge. In a second step, a 20% solution of aqueous ion-pairing reagent in acetonitrile was applied. 13 N-oxytocin is eluted from the cartridge. 13 The eluate containing N-oxytocin was collected, diluted, loaded onto a second C18 Sep-pak cartridge, and washed with HO to remove acetonitrile and ion-pairing reagent. Finally, EtOH was used to remove the eluate. 13 N-oxytocin was eluted from the cartridge and purified. 13 The eluate containing N-oxytocin was collected. [Table 1]

Claims

1. A 13 N-labeled oxytocin peptide comprising the amino acid sequence of SEQ ID NO:

1.

2. The 13 N-labeled oxytocin peptide of claim 1, wherein one constituent 14 N atom of the oxytocin peptide is replaced with a 13 N radionuclide.

3. The 13 N-labeled oxytocin peptide of claim 1 or 2, wherein one residue of the oxytocin peptide is modified to include a moiety containing a 13 N radionuclide.

4. The 13 N-labeled oxytocin peptide of claim 3, wherein one residue of the oxytocin peptide is modified to contain 13 NH 2 .

5. A 13 N-labeled oxytocin peptide according to any one of claims 1 to 4, comprising a single 13 N radionuclide.

6. A 13N-labeled oxytocin peptide described in any one of claims 1 to 5, comprising a 13N radionuclide at a) the glutamine residue at position 4 (SEQ ID NO: 2), b) the asparagine residue at position 5 (SEQ ID NO: 3), or c) the glycine residue at position 9 (SEQ ID NO: 4).

7. The 13 N-labeled oxytocin peptide of formula (IV): 【Chemistry 1】 2. The 13 N-labeled oxytocin peptide of claim 1 , which is a compound of formula:

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