HK1252813B - Therapeutic compounds and compositions for treating social disorders and substance use disorders - Google Patents

Description

Therapeutic compounds and compositions for treating social disorders and substance use disorders

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims priority from Australian provisional patent application 2015902659 filed on 6 July 2015, the contents of which are incorporated herein by reference.

Technical Field

The present disclosure relates to compounds, compositions and methods for treating neurological and psychiatric diseases characterized by fundamental disruptions in social behavior and substance use disorders. In specific embodiments, the present disclosure relates to the use of compounds of formula (I) or salts and prodrugs of compounds of formula (I) to treat neurological and psychiatric diseases and substance use disorders.

Background Art

Many psychiatric disorders are characterized by fundamental disruptions in social behavior. Common examples include autism spectrum disorder (ASD) and social anxiety disorder (SAD). In addition, some diseases have secondary symptoms of social withdrawal, including schizophrenia, major depressive disorder (MDD), and substance use disorders. There are currently no approved medications that directly target deficits in social functioning, and drug treatments for these disorders, if any, have only limited efficacy.

Recent preclinical studies have shown that the neuropeptide oxytocin has significant therapeutic potential in stimulating social behavior and reducing drug and alcohol self-administration. However, oxytocin itself has poor blood-brain barrier permeability, negligible oral bioavailability, and a short in vivo half-life. Due to these issues, clinical studies using intranasal oxytocin have yielded only modest benefits in clinical populations. Therefore, small molecules capable of stimulating the brain oxytocin system with greater efficacy may have broad therapeutic applications in the aforementioned disorders.

These disorders are among the most prevalent and contribute significantly to the global burden of disease. A recent study by the Centers for Disease Control estimated the prevalence of autism in 1 in 110 8-year-olds. SAD is the second most common anxiety disorder, with the National Institutes of Health (NIH) estimating that approximately 6.8% of American adults suffer from it.

The World Health Organization (WHO) estimates that schizophrenia affects approximately 24 million people worldwide, the highest prevalence of any chronic illness. The WHO also estimates that in 2012, over 3 million people died worldwide from alcohol abuse (5.9% of all deaths). In developed countries, psychotropic substance abuse accounts for 33.4% of all life years lost among men.

The idea of specifically targeting the neural underpinnings of social behavior represents a paradigm shift in the treatment of these disorders. For example, the use of antipsychotics (e.g., risperidone) in ASD is intended to suppress aggressive and challenging behaviors rather than stimulate prosocial behaviors. The use of antidepressants (e.g., paroxetine or venlafaxine) in SAD assumes that depressed mood and generalized anxiety are the primary contributors to social withdrawal in SAD and indirectly affect social anxiety by improving mood and reducing global anxiety. Treatment of schizophrenia with antipsychotics (e.g., olanzapine or aripiprazole) attempts to control positive symptoms and, to some extent, neurocognitive impairment but does not directly address the pervasive social withdrawal seen in the chronic condition.

Current addiction treatments offer alternative versions of drugs of abuse (e.g., methadone, buprenorphine, or varenicline) to control cravings, or treatments that can reduce cravings through largely unknown mechanisms (e.g., acamprosate, naltrexone, baclofen, or ondansetron). These current pharmacotherapy treatments for addiction have limited effectiveness, and none targets the debilitating social dysfunction caused by these disorders. While there are some psychological treatments for these disorders, their success has also been limited.

The disclosed compounds of formula (I) and their salts and prodrugs have been developed to directly target social dysfunction by acting on brain regions known to be involved in the acute and long-term regulation of social behavior in an attempt to produce a long-term shift in social motivation that may far exceed the duration of drug treatment, representing a "change of state." In other words, these compounds have been developed to recalibrate an individual's social behavior to a higher level through treatment. In terms of treating drug abuse, the inventors believe that a sustained increase in social motivation may redirect behavior toward social rewards rather than a destructive cycle of drug-seeking behavior.

Any discussion of documents, acts, materials, devices, articles or the like in this specification is not to be taken as an admission that any or all of these matters form part of the prior art base or were general knowledge in the field relevant to the present disclosure as it existed before the priority date of each claim of this application.

Summary of the Invention

The first aspect of the present invention provides a compound of formula (I), or a salt thereof, or a prodrug thereof:

in:

V is NH, CH ₂ or a direct bond;

W is NH, CH ₂ or a direct bond;

X is NH, CH ₂ or a direct bond;

Y is NH, CH ₂ or a direct bond;

Z is selected from: NH, O, S, S(O), SO ₂ or a direct bond;

R ¹ is selected from H or C(O)R ⁴ ;

R ² is selected from: H, OH, halogen, optionally substituted C _1-5 alkyl or optionally substituted OC _1-5 alkyl;

R ³ is selected from: H, OH, halogen, optionally substituted C _1-5 alkyl or optionally substituted OC _1-5 alkyl;

R ⁴ is an optionally substituted C _1-5 alkyl group;

m is 0 or 1;

n is 0 or 1;

p is 0 or 1; and

q is 0 or 1.

The second aspect of the present invention provides a pharmaceutical composition comprising the pharmaceutically acceptable compound according to the first aspect of the present invention, or a pharmaceutically acceptable salt thereof, or a prodrug thereof and a pharmaceutically acceptable carrier, diluent or excipient.

The third aspect of the present invention provides a method for treating or preventing a condition in a subject, the method comprising the steps of: administering the pharmaceutically acceptable compound described in the first aspect of the present invention, or a pharmaceutically acceptable salt thereof, or a prodrug thereof; or the pharmaceutical composition described in the second aspect of the present invention.

The fourth aspect of the present invention provides a use of the pharmaceutically acceptable compound described in the first aspect of the present invention, or a pharmaceutically acceptable salt thereof, or a prodrug thereof; or the pharmaceutical composition described in the second aspect of the present invention, for treating or preventing a condition in a subject.

The fourth aspect of the present invention provides a use of the pharmaceutically acceptable compound described in the first aspect of the present invention, or a pharmaceutically acceptable salt thereof, or a prodrug thereof; or the pharmaceutical composition described in the second aspect of the present invention, for preparing a drug for treating or preventing a condition in a subject.

BRIEF DESCRIPTION OF THE DRAWINGS

Figure 1 (A) shows the results of the social interaction test in male Hooded Wistar rats given different doses of SOC-1 intraperitoneally (IP); (B) shows the residual prosocial effects of different doses of SOC-1 on Hooded Wistar rats in the social interaction test; (C) shows the comparison results of adjacent lying in the social interaction test, Hooded Wistar rats were given vehicle (VEH); oxytocin (OT) (0.5 mg / kg); arginine vasopressin (AVP) (0.005 mg / kg); 3,4-methylenedioxy-methamphetamine (MDMA) (5 mg / kg); WAY 267,464 (100 mg / kg); or SOC-1 (5 mg / kg).

Figure 2 (A) shows the results of a social preference test, in which SOC-1 (5 mg/kg IP) enhanced rats' natural preference for an unfamiliar live rat (solid part of the bar) rather than a toy rat (patterned part of the bar); - (B) shows the selective effect of orally administered SOC-1 (3 mg/kg) on intravenous cocaine self-administration in rhesus monkeys.

FIG3 Cumulative rate of cocaine self-administration in macaques administered SOC-1.

FIG4 shows the concentration-time curves of SOC-1 administered at the same dose (5 mg/kg) by intraperitoneal injection (IP) or oral administration (PO).

Figure 5. Stability of SOC-1 incubated in human plasma at body temperature.

FIG6 Fos immunohistochemistry studies testing activation of oxytocin-containing cells in the supraoptic nucleus (SON) of the hypothalamus of male Wistar rats, with: vehicle ( (A) ); oxytocin at a concentration of 1 mg/kg ( - (B) ); and SOC-1 at a concentration of 5 mg/kg ( (C) ).

Figure 7 - Body temperature was monitored in Wistar rats administered vehicle, SOC-1 (5 mg/kg), or SOC-1 and the V1A receptor antagonist SR49059 (10 mg/kg).

Figure 8 - Heart rate was monitored in Wistar rats administered vehicle, SOC-1 (5 mg/kg), or SOC-1 and the V1A receptor antagonist SR49059 (10 mg/kg).

Figure 9 Effects of SOC-1 on methamphetamine self-administration in rats under a progressive ratio scale. * indicates p < 0.05 relative to vehicle.

Figure 10 Effects of SOC-1 on lever pressing for methamphetamine self-administration in rats under a progressive ratio schedule. * indicates p < 0.05 relative to vehicle.

Figure 11 Effects of SOC-1 on locomotor activity in rats self-administering methamphetamine under a progressive ratio scale. * indicates p < 0.05 relative to vehicle.

FIG12 Effects of vehicle, SOC-1, and the combination of SOC-1 and C25 or SR49059 on active lever pressing during methamphetamine-induced reinstatement. # indicates p < 0.05, main effect of day, & indicates p < 0.05, main effect of treatment, and * indicates p < 0.05 relative to vehicle + METH condition.

FIG13 Effects of vehicle, SOC-1, and the combination of SOC-1 and C25 or SR49059 on locomotor activity during methamphetamine-induced reinstatement. # indicates p < 0.05, main effect of day, & indicates p < 0.05, main effect of treatment, and * indicates p < 0.05 relative to vehicle + METH condition.

Figure 14. Mean body weight values of Sprague Dawley rats consuming SOC-1.

Figure 15 is a graph of mean (+ standard error of the mean (SEM)) plasma SOC-1 concentration versus time in Sprague Dawley rats.

Figure 16 shows the mean body weight values of Sprague Dawley rats administered SOC-1 on study days.

Figure 17 Total fluid delivery for alcohol-intoxicated baboons during the study period versus various concentrations of SOC-1 administered to the baboons and overall mean values.

Figure 18 Alcohol intake in baboons and mean values during the morning phase of baboons given different doses of SOC-1.

Figure 19 Alcohol intake in baboons and mean values during the afternoon phase of baboons given different doses of SOC-1.

FIG20 shows the results of autism spectrum disorder research using BALB/C mice administered SOC-1.

Detailed Description of the Invention

In this specification, the term "comprise" or variations such as "comprises" or "comprising" are understood to mean the inclusion of stated elements, integers or steps, or combinations of elements, integers or steps, but not the exclusion of any other elements, integers or steps, or combinations of elements, integers or steps.

With respect to the definitions provided herein, unless otherwise stated or implied from the context, the defined terms and phrases include the meanings provided. Unless otherwise expressly stated or apparent from the context, the following terms and phrases do not exclude the meanings already assigned to the terms or phrases by persons skilled in the relevant art. These definitions are provided to aid in the description of particular embodiments and are not intended to limit the claimed invention, as the scope of the invention is limited solely by the claims. Furthermore, unless the context requires otherwise, the singular shall include the plural and the plural shall include the singular.

In this specification, various aspects and components of the present invention may be presented in the form of ranges. The range format is included for convenience and should not be interpreted as a fixed limitation on the scope of the present invention. Therefore, unless expressly indicated, the description of a range should be considered to have specifically disclosed all possible subranges and individual numerical values within the range. For example, the description of a range from 1 to 5 should be considered to have specifically disclosed subranges, such as from 1 to 3, 1 to 4, 1 to 5, 2 to 4, 2 to 5, from 3 to 5, etc., as well as individual and partial numbers within the range, such as 1, 2, 3, 4, 5, 5.5 and 6. This applies regardless of the width of the disclosed range. Where specific values are required, these will be indicated in the specification.

As used herein, the term "C _1-5 alkyl" used alone or in compound terms refers to a monovalent straight or branched hydrocarbon group having 1 to 5 carbon atoms. As understood by those skilled in the art, the term "C _1-5 alkyl" refers to an alkyl group having 1, 2, 3, 4 or 5 carbon atoms or a range comprising any two of these integers, including 1-2, 1-3, 1-4, 1-5, 2-3, 2-4, 2-5, 3-4, 3-5 and 4-5. Suitable alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, n-pentyl, neopentyl, isopentyl and tert-pentyl. The _{C 1-4} alkyl group may be optionally substituted with one or more substituents. The substituents may be at any position along the carbon chain. Suitable substituents include, but are not limited to, OH, NH ₂ , halogen, NH(C _1-5 alkyl), N(C _1-5 alkyl) ₂ , CN, NO ₂ , CO ₂ H, or OC _1-5 alkyl.

As used herein, the term "OC _1-5 alkyl" used alone or in compound terms refers to an alkoxy group having 1 to 5 carbon atoms. As understood by those skilled in the art, the term "OC _1-5 alkyl" refers to an alkoxy group having 1, 2, 3, 4 or 5 carbon atoms or a range containing any two of these integers, including 1-2, 1-3, 1-4, 1-5, 2-3, 2-4, 2-5, 3-4, 3-5 and 4-5. Suitable OC _1-5 alkyl groups include, but are not limited to, methoxy, ethoxy, propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, n-pentoxy, neopentoxy, isopentoxy and tert-pentoxy. C _1-5 alkyl groups may be optionally substituted with one or more substituents. The substituents may be at any position along the carbon chain. Suitable substituents include, but are not limited to, OH, NH ₂ , halogen, NH(C _1-5 alkyl), N(C _1-5 alkyl) ₂ , CN, NO ₂ , CO ₂ H or OC _1-5 alkyl groups.

As used herein, the term "halo" or "halogen" refers to fluoro (fluorine), chloro (chlorine), bromo (bromine), or iodo (iodine).

A compound of formula (I) or a salt or a prodrug thereof:

Disclosed herein are compounds of formula (I), or salts thereof, or prodrugs thereof,

in:

V is NH, CH ₂ or a direct bond;

W is NH, CH ₂ or a direct bond;

X is NH, CH ₂ or a direct bond;

Y is NH, CH ₂ or a direct bond;

Z is selected from: NH, O, S, S(O), SO ₂ or a direct bond;

R ¹ is selected from H or C(O)R ⁴ ;

R ² is selected from: H, OH, halogen, optionally substituted C _1-5 alkyl or optionally substituted OC _1-5 alkyl;

R ³ is selected from: H, OH, halogen, optionally substituted C _1-5 alkyl or optionally substituted OC _1-5 alkyl;

R ⁴ is an optionally substituted C _1-5 alkyl group;

m is 0 or 1;

n is 0 or 1;

p is 0 or 1; and

q is 0 or 1.

Disclosed herein are compounds of formula (Ia), or salts thereof, or prodrugs thereof,

in:

Z is selected from: NH, O, S, S(O) or SO ₂ ;

R ¹ is selected from: H or C(O)R ⁴ ;

R ² is selected from: H, OH, halogen, optionally substituted C _1-5 alkyl or optionally substituted OC _1-5 alkyl;

R ³ is selected from the group consisting of: H, OH, halogen, optionally substituted C _1-5 alkyl, or optionally substituted OC _1-5 alkyl; and

R ⁴ is an optionally substituted C _1-5 alkyl group.

Disclosed herein are compounds of formula (Ib), or salts thereof, or prodrugs thereof,

in:

R ¹ is selected from H or C(O)R ⁴ ;

R ² is selected from: H, OH, halogen, optionally substituted C _1-5 alkyl or optionally substituted OC _1-5 alkyl;

R ³ is selected from: H, OH, halogen, optionally substituted C 1-5 alkyl or optionally substituted OC _1-5 alkyl; and

R ⁴ is an optionally substituted C _1-5 alkyl group.

Disclosed herein are compounds of formula (Ic), or salts thereof, or prodrugs thereof,

in:

Z is selected from: NH, O, S, S(O) or SO ₂ ;

R ¹ is selected from H or C(O)R ⁴ ;

R ² is selected from: H, OH, halogen, optionally substituted C _1-5 alkyl or optionally substituted OC _1-5 alkyl;

R ³ is selected from the group consisting of: H, OH, halogen, optionally substituted C _1-5 alkyl, or optionally substituted OC _1-5 alkyl; and

R ⁴ is an optionally substituted C _1-5 alkyl group.

Disclosed herein are compounds of formula (Id), or salts thereof, or prodrugs thereof,

in:

Z is selected from: NH, O, S, S(O) or SO ₂ ;

R ¹ is selected from H or C(O)R ⁴ ;

R ² is selected from: H, OH, halogen, optionally substituted C _1-5 alkyl or optionally substituted OC _1-5 alkyl;

R ³ is selected from the group consisting of: H, OH, halogen, optionally substituted C _1-5 alkyl, or optionally substituted OC _1-5 alkyl; and

R ⁴ is an optionally substituted C _1-5 alkyl group.

Disclosed herein are compounds of formula (Ie), or salts thereof, or prodrugs thereof,

in:

Z is selected from: NH, O, S, S(O) or SO ₂ ;

R ¹ is selected from H or C(O)R ⁴ ;

R ² is selected from: H, OH, halogen, optionally substituted C _1-5 alkyl or optionally substituted OC _1-5 alkyl;

R ³ is selected from the group consisting of: H, OH, halogen, optionally substituted C _1-5 alkyl, or optionally substituted OC _1-5 alkyl; and

R ⁴ is an optionally substituted C _1-5 alkyl group.

Disclosed herein are compounds of formula (If), or salts thereof, or prodrugs thereof,

in:

Z is selected from: NH, O, S, S(O) or SO2;

R ¹ is selected from H or C(O)R ⁴ ;

R ² is selected from: H, OH, halogen, optionally substituted C _1-5 alkyl or optionally substituted OC _1-5 alkyl;

R ³ is selected from the group consisting of: H, OH, halogen, optionally substituted C _1-5 alkyl, or optionally substituted OC _1-5 alkyl; and

R ⁴ is an optionally substituted C _1-5 alkyl group.

In one embodiment, the compound of formula (I) is a compound of formula (Ia), or a salt thereof, or a prodrug thereof.

In one embodiment, the compound of formula (I) is a compound of formula (Ib), or a salt thereof, or a prodrug thereof. In one embodiment, the compound of formula (I) is a compound of formula (Ic), or a salt thereof, or a prodrug thereof. In one embodiment, the compound of formula (I) is a compound of formula (Id), or a salt thereof, or a prodrug thereof. In one embodiment, the compound of formula (I) is a compound of formula (Ie), or a salt thereof, or a prodrug thereof. In one embodiment, the compound of formula (I) is a compound of formula (If), or a salt thereof, or a prodrug thereof.

In one embodiment, V is NH.

In one embodiment, V is CH ₂ .

In one embodiment, V is a direct bond.

In one embodiment, W is NH.

In one embodiment, W is CH ₂ .

In one embodiment, W is a direct bond.

In one embodiment, X is NH.

In one embodiment, X is CH ₂ .

In one embodiment, X is a direct bond.

In one embodiment, Y is NH.

In one embodiment, Y is CH ₂ .

In one embodiment, Y is a direct bond

In one embodiment, Z is NH.

In one embodiment, Z is O.

In one embodiment, Z is S.

In one embodiment, Z is S(O).

In one embodiment, Z is SO ₂ .

In one embodiment, Z is a direct bond.

In one embodiment, R ¹ is hydrogen.

In one embodiment, R ¹ is C(O)R ^4. For example, R ⁴ can be an optionally substituted C _1-5 alkyl selected from the group consisting of methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, n-pentyl, neopentyl, isopentyl, and tert-pentyl. In one embodiment, R ⁴ is an optionally substituted methyl.

In one embodiment, ^R2 is hydrogen.

In one embodiment, ^R2 is hydroxy.

In one embodiment, R ² is halogen. For example, in one embodiment, R ² is fluorine. In another embodiment, R ² is chlorine.

In one embodiment, ^R is an optionally substituted C _1-5 alkyl group. For example, ^R can be an optionally substituted C _1-5 alkyl group selected from the group consisting of methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, n-pentyl, neopentyl, isopentyl, and tert-pentyl. In one embodiment, ^R is an optionally substituted methyl group.

In one embodiment, ^R is an optionally substituted _0-1-5 alkyl group. For example, ^R can be an optionally substituted _0-1-5 alkyl group selected from the group consisting of methoxy, ethoxy, propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, n-pentoxy, neopentoxy, isopentoxy, and tert-pentoxy. In one embodiment, ^R is an optionally substituted methoxy group.

In one embodiment, R ³ is hydrogen.

In one embodiment, R ³ is hydroxy.

In one embodiment, R ³ is halogen. For example, in one embodiment, R ³ is fluorine. In another embodiment, R ³ is chlorine.

In one embodiment, ^R is an optionally substituted C _1-5 alkyl group. For example, ^R can be an optionally substituted C _1-5 alkyl group selected from the group consisting of methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, n-pentyl, neopentyl, isopentyl, and tert-pentyl. In one embodiment, ^R is an optionally substituted methyl group.

In one embodiment, ^R is an optionally substituted _0-1-5 alkyl group. For example, ^R can be an optionally substituted _0-1-5 alkyl group selected from the group consisting of methoxy, ethoxy, propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, n-pentoxy, neopentoxy, isopentoxy, and tert-pentoxy. In one embodiment, ^R is an optionally substituted methoxy group.

In one embodiment, the compound of formula (I) is selected from:

or a salt thereof, or a prodrug thereof.

In one embodiment, the compound of formula (I) is selected from:

or a salt thereof, or a prodrug thereof.

In one embodiment, the compound of formula (I) is selected from:

or a salt thereof, or a prodrug thereof.

In one embodiment, the compound of formula (I) is selected from:

or a salt thereof, or a prodrug thereof.

In one embodiment, the compound of formula (I) is selected from:

or a salt thereof, or a prodrug thereof.

In one embodiment, the compound of formula (I), or a salt thereof, or a prodrug thereof is pharmaceutically acceptable.

In one embodiment, the compound of formula (I) is a compound of formula (Ia), or a salt thereof, or a prodrug thereof, and is pharmaceutically acceptable.

In one embodiment, the compound of formula (I) is a compound of formula (Ib), or a salt thereof, or a prodrug thereof, and is pharmaceutically acceptable.

In one embodiment, the compound of formula (I) is a compound of formula (Ic), or a salt thereof, or a prodrug thereof, and is pharmaceutically acceptable.

In one embodiment, the compound of formula (I) is a compound of formula (Id), or a salt thereof, or a prodrug thereof, and is pharmaceutically acceptable.

In one embodiment, the compound of formula (I) is a compound of formula (Ie), or a salt thereof, or a prodrug thereof, and is pharmaceutically acceptable.

In one embodiment, the compound of formula (I) is a compound of formula (If), or a salt thereof, or a prodrug thereof, and is pharmaceutically acceptable.

The compound of formula (I), or its salt, or its prodrug may be pharmaceutically acceptable, but it should be understood that non-pharmaceutically acceptable compounds, their salts and their prodrugs also fall within the scope of the present invention because they are used as intermediates for preparing pharmaceutically acceptable compounds, their salts and their prodrugs.

In one embodiment, the compound of formula (I) is a salt, such as a pharmaceutically acceptable salt.

In one embodiment, the compound of formula (I) is a compound of formula (Ia) and is a salt, such as a pharmaceutically acceptable salt.

In one embodiment, the compound of formula (I) is a compound of formula (Ib) and is a salt, such as a pharmaceutically acceptable salt.

In one embodiment, the compound of formula (I) is a compound of formula (Ic) and is a salt, such as a pharmaceutically acceptable salt.

In one embodiment, the compound of formula (I) is a compound of formula (Id) and is a salt, such as a pharmaceutically acceptable salt.

In one embodiment, the compound of formula (I) is a compound of formula (Ie) and is a salt, such as a pharmaceutically acceptable salt.

In one embodiment, the compound of formula (I) is a compound of formula (If) and is a salt, such as a pharmaceutically acceptable salt.

Suitable pharmaceutically acceptable salts include, but are not limited to, salts of pharmaceutically acceptable inorganic acids such as hydrochloric, sulfuric, phosphoric, nitric, carbonic, boric, sulfamic, and hydrobromic acids; or salts of pharmaceutically acceptable organic acids such as acetic, propionic, butyric, tartaric, maleic, hydroxymaleic, fumaric, isethionic, malic, citric, lactic, mucic, gluconic, benzoic, succinic, oxalic, phenylacetic, methanesulfonic, toluenesulfonic, benzenesulfonic, salicylic, sulfanilic, aspartic, glutamic, edetic, stearic, palmitic, oleic, lauric, pantothenic, tannic, ascorbic, and valeric acids.

Basic salts include, but are not limited to, salts with pharmaceutically acceptable cations such as sodium, potassium, lithium, calcium, magnesium, zinc, ammonium, and alkylammonium; salts with triethylamine; alkoxyammonium salts, such as salts with ethanolamine; and salts with ethylenediamine, choline, or amino acids such as arginine, lysine, or histidine. General information on the types of pharmaceutically acceptable salts and their forms is known to those skilled in the art and is described in general textbooks such as "Handbook of Pharmaceutical Salts" (P.H. Stahl, C.G. Wermuth, 1st edition, 2002, Wiley-VCH).

The basic nitrogen-containing groups of formula (I) (or the basic nitrogen-containing groups of formula (Ia), (Ib), (Ic), (Id), (Ie) or (If)) can be quaternized with lower alkyl halides such as chlorides, bromides and iodides of methyl, ethyl, propyl and butyl, dialkyl sulfates such as dimethyl sulfate and diethyl sulfate, or other agents known in the art.

In one embodiment, the compound of formula (I) is a salt of a compound selected from the group consisting of:

In one embodiment, the compound of formula (I) is a salt of a compound selected from the group consisting of:

In one embodiment, the compound of formula (I) is a salt of a compound selected from the group consisting of:

In one embodiment, the salt of the compound of formula (I) is a salt of:

In another embodiment, the salt of the compound of formula (I) is a salt of:

In one embodiment, the compound of formula (I) is a hydrochloride salt.

In one embodiment, the compound of formula (I) is a compound of formula (Ia) in the form of a hydrochloride salt, such as a pharmaceutically acceptable hydrochloride salt.

In one embodiment, the compound of formula (I) is a compound of formula (Ib) in the form of a hydrochloride salt, such as a pharmaceutically acceptable hydrochloride salt.

In one embodiment, the compound of formula (I) is a compound of formula (Ic) in the form of a hydrochloride salt, such as a pharmaceutically acceptable hydrochloride salt.

In one embodiment, the compound of formula (I) is a compound of formula (Id) in the form of a hydrochloride salt, such as a pharmaceutically acceptable hydrochloride salt.

In one embodiment, the compound of formula (I) is a compound of formula (Ie) in the form of a hydrochloride salt, such as a pharmaceutically acceptable hydrochloride salt.

In one embodiment, the compound of formula (I) is a compound of formula (If) in the form of a hydrochloride salt, such as a pharmaceutically acceptable hydrochloride salt.

In one embodiment, the compound of formula (I) is a hydrochloride salt of a compound selected from the group consisting of:

In one embodiment, the compound of formula (I) is a hydrochloride salt of a compound selected from the group consisting of:

In one embodiment, the compound of formula (I) is a hydrochloride salt of a compound selected from the group consisting of:

For example, in one embodiment, the salt of the compound of formula (I) is the hydrochloride salt of:

In another embodiment, the salt of the compound of formula (I) is the hydrochloride of the following compound:

In one embodiment, the compound of formula (I) is a prodrug.

In one embodiment, the compound of formula (Ia) is a prodrug.

In one embodiment, the compound of formula (Ib) is a prodrug.

In one embodiment, the compound of formula (Ic) is a prodrug.

In one embodiment, the compound of formula (Id) is a prodrug.

In one embodiment, the compound of Formula (Ie) is a prodrug.

In one embodiment, the compound of formula (If) is a prodrug.

Prodrugs include compounds in which an amino acid residue or a polypeptide chain of two or more (e.g., 2, 3, or 4) amino acid residues is covalently linked to a free amino group, a hydroxyl group, and a carboxylic acid group of a compound of formula (I). Amino acid residues include 20 naturally occurring amino acids typically represented by three letter symbols, and also include: 4-hydroxyproline, hydroxylysine, dehydroinosine, isomeric adamantane, 3-methylhistidine, norvaline, β-alanine, γ-aminobutyric acid, citrulline, homocysteine, homoserine, ornithine, and methionine sulfone. Prodrugs also include compounds in which carbonates, carbamates, amides, and alkyl esters can be covalently linked to the above-mentioned substituents of formula (I) via a carbonyl carbon prodrug side chain. Prodrugs also include phosphate derivatives of a compound of formula (I) (e.g., a salt of an acid, acid, or ester) linked to a free hydroxyl group of a compound of formula (I) via a phosphorus-oxygen bond.

Pharmaceutical composition

The present invention discloses a pharmaceutical composition comprising a compound of formula (I), or a salt thereof, or a prodrug thereof, and a pharmaceutically acceptable carrier, diluent or excipient. In one embodiment, the compound of formula (I), or a salt thereof, or a prodrug thereof is pharmaceutically acceptable.

As used herein, "pharmaceutically acceptable carriers, diluents, or excipients" include excipients or agents that are physiologically compatible and not deleterious to the compounds described herein or their uses, such as solvents, diluents, dispersion media, coatings, antibacterial and antifungal agents, isotonicity regulators, and absorption delaying agents. The use of such carriers and agents to prepare compositions of pharmaceutically active substances is well known in the art, see, for example, Remington: The Science and Practice of Pharmacy (21st ed.; Lippincott Williams & Wilkins: Philadelphia, Pennsylvania, 2005).

The pharmaceutically acceptable composition may be diluted before use. Suitable diluents may be selected from, for example, Ringer's solution, Hartmann's solution, glucose solution, saline solution, and sterile water for injection.

As used herein, the term "composition" is intended to encompass a product comprising the specified ingredients in the specified amounts, as well as any product which results, directly or indirectly, from combination of the specified ingredients in the specified amounts.

As used herein, "pharmaceutically acceptable" means that the compound of formula (I), or its salt, or its prodrug and the carrier, diluent, or excipient must be compatible with the other ingredients of the formulation and not harmful to the recipient. The compound of formula (I), or its salt, or its prodrug should be able to contact the recipient's tissues without excessive toxicity, irritation, allergic reaction, or other potential complications, commensurate with a reasonable benefit/risk ratio determined by a skilled medical professional or veterinarian. In addition, the compound of formula (I), or its salt, or its prodrug should be compatible with the carrier and/or excipient in any composition that is delivered to an individual, such as an animal or a human, as a drug.

It should be understood that in some cases, the compound of formula (I), or its salt, or its prodrug is non-pharmaceutically acceptable, and these fall within the scope of the present invention. However, in order to be used in pharmaceutical preparations, in some embodiments, the compound of formula (I), or its salt, or its prodrug is a pharmaceutically acceptable compound. The non-pharmaceutically acceptable compound of formula (I), or its salt, or its prodrug can be used to prepare a pharmaceutically acceptable compound of formula (I), or its salt, or its prodrug.

The compositions of the present invention may contain other therapeutic agents as described below, and can be formulated, for example, using conventional solid or liquid carriers or diluents and pharmaceutical additives suitable for the desired form (e.g., excipients, binders, preservatives, stabilizers, flavorings, etc.), and administered according to, for example, techniques well known in the field of pharmaceutical preparations.

The compounds of formula (I) as defined herein, or salts thereof, or prodrugs thereof, may be administered by any suitable route, for example parenterally, such as by subcutaneous, intraperitoneal, intravenous, intramuscular or intracerebral injection or infusion techniques (for example, as a sterile injectable aqueous or non-aqueous solution or suspension).

Pharmaceutical preparations include those for oral, rectal, nasal, topical (including buccal and sublingual), parenteral (including intramuscular, intraperitoneal, subcutaneous and intravenous) administration, or in forms suitable for administration by inhalation or spray. The compound of formula (I), or a salt thereof, or a prodrug thereof, together with conventional adjuvants, carriers or diluents, can be stored in the form of a pharmaceutical composition and a unit dose thereof, and can be in the form of a solid such as a tablet or a filled capsule, or a liquid such as a solution, suspension, emulsion, elixir or a capsule filled with the same contents, all for oral administration, or in the form of a sterile injection for parenteral (including subcutaneous) use.

The pharmaceutical composition for administering the compound of the present invention can be conveniently present in the form of dosage units and can be prepared by any method known in the pharmaceutical field. All methods include the step of combining an active ingredient such as a compound of formula (I), or a salt thereof, or a prodrug thereof with a carrier consisting of one or more auxiliary ingredients. Typically, the pharmaceutical composition is prepared by uniformly and closely combining an active ingredient such as a compound of formula (I), or a salt thereof, or a prodrug thereof with a liquid carrier or a thin solid carrier or both, and then (if necessary) the product is shaped into a desired preparation. In the pharmaceutical composition, the content of the active target compound is enough to produce the desired effect on the process or condition of the disease.

The pharmaceutical composition can be in the form of a sterile injection water or oil suspension. The suspension can be prepared according to known techniques using those suitable dispersants or wetting agents and suspending agents mentioned above. Sterile injection preparations can also be sterile injection solutions or suspensions in non-toxic parenteral acceptable diluents or solvents, such as solutions in 1,3-butanediol. Among acceptable carriers and solvents, water, Ringer's solution, and isotonic sodium chloride solution can be used. In addition, sterile and solid phase oils are usually used as solvents or suspension media. For this purpose, any mild solid phase oil can be used, including synthetic monoglycerides or diglycerides. In addition, fatty acids such as oleic acid can be used to prepare injectable preparations.

The pharmaceutical compositions and methods described herein may further comprise other therapeutically active compounds commonly used to treat the diseases or conditions described herein. Based on conventional pharmaceutical principles, one of ordinary skill in the art can select suitable agents for combination therapy. Combinations of therapeutic agents can act synergistically to achieve the treatment or prevention of the various diseases or conditions described herein. Using this approach, therapeutic effects can be achieved with lower doses of each agent, thereby reducing the likelihood of adverse side effects.

When other therapeutic agents are used in combination with the compounds of formula (I), or salts thereof, or prodrugs thereof, they may be used in the amounts described in the Physician Desk Reference (PDR), or in other ways determined by one of ordinary skill in the art.

However, it should be understood that the specific dosage level and frequency of administration for any particular patient may vary depending on various factors, including the activity of the specific compound employed, the metabolic stability and duration of action of the compound, the age, body weight, general health, sex, diet, mode and time of administration, rate of excretion, drug combination, the severity of the specific condition, and the subject being treated.

Treatment

Disclosed herein are methods for treating or preventing a condition in a subject, comprising administering to the subject a pharmaceutically acceptable amount of a compound of formula (I) of the present invention, or a pharmaceutically acceptable salt thereof, or a prodrug thereof, or a pharmaceutical composition described herein.

Also disclosed herein is the use of a pharmaceutically acceptable compound of formula (I), or a pharmaceutically acceptable salt thereof, or a prodrug thereof, or the pharmaceutical composition of the present invention for treating or preventing a condition in a subject.

Also disclosed herein is the use of a pharmaceutically acceptable compound of formula (I), or a pharmaceutically acceptable salt thereof, or a prodrug thereof in the preparation of a medicament for treating or preventing a condition in a subject.

In one embodiment, the compound of formula (I), or a salt thereof, or a prodrug thereof is used as a medicament.

In one embodiment, the compound of formula (Ia), or a salt thereof, or a prodrug thereof is used as a medicament.

In one embodiment, the compound of formula (Ib), or a salt thereof, or a prodrug thereof is used as a medicament.

In one embodiment, the compound of formula (Ic), or a salt thereof, or a prodrug thereof is used as a medicament.

In one embodiment, the compound of formula (Id), or a salt thereof, or a prodrug thereof is used as a medicament.

In one embodiment, the compound of formula (Ie), or a salt thereof, or a prodrug thereof is used as a medicament.

In one embodiment, the compound of formula (If), or a salt thereof, or a prodrug thereof is used as a medicament.

The compound of formula (I), or a salt thereof, or a prodrug thereof can be provided in an "effective amount," for example, when a suitable compound is added to a pharmaceutical composition. An "effective amount" is considered to be that amount of a compound that, when administered to a researcher, veterinarian, medical doctor, or other clinician administering the compound or a composition containing the compound, will elicit the desired biological or medical response in a tissue, system, animal, or human.

An "effective amount" depends on many factors, including the therapeutic efficacy of the particular compound. The subject's weight and age are also factors considered by those skilled in the art when determining the amount of compound a subject should receive.

The terms "administration" and/or "administering" a compound should be understood to mean providing a compound of Formula (I) (or Formula (Ia), or Formula (Ib), or Formula (Ic), a compound of Formula (Id), or Formula (Ie), or Formula (If)), or a salt thereof, or a prodrug thereof to a subject in need of treatment.

The recipient of the compound of formula (I), or a pharmaceutically acceptable salt thereof, or a prodrug thereof; or a pharmaceutical composition comprising the compound of formula (I), or a pharmaceutically acceptable salt thereof, or a prodrug thereof, can be a human, including male or female.

Alternatively, the subject of the compound of formula (I), or a pharmaceutically acceptable derivative thereof, or a salt thereof, or a prodrug thereof; or a pharmaceutical composition comprising a compound of formula (I), or a pharmaceutically acceptable derivative thereof, or a salt thereof, or a prodrug thereof may also be a non-human animal. "Non-human animals" or "non-human animals" refers to the animal kingdom, excluding humans, including vertebrates and invertebrates, male or female, including: warm-blooded animals, including mammals (including but not limited to primates, dogs, cats, cattle, pigs, sheep, goats, rats, guinea pigs, horses or other cattle, sheep, horses, dogs, felines, rodents or murines), birds, insects, reptiles, fish and amphibians.

The subjects of the compounds and pharmaceutically acceptable compositions are interchangeably referred to herein as "patient," "subject," "individual," and "subject." These four terms are used interchangeably to refer to any human or animal as defined herein (unless otherwise indicated).

In one embodiment, the compound of formula (I), or a salt thereof, or a prodrug thereof; or a pharmaceutical composition comprising the compound of formula (I), or a salt thereof, or a prodrug thereof is used to stimulate prosocial behavior in a subject.

In one embodiment: the compound of formula (I), or a salt thereof, or a prodrug thereof; or a pharmaceutical composition comprising the compound of formula (I), or a salt thereof, or a prodrug thereof is used to provide acute and long-term regulation of a subject's social behavior.

In one embodiment: a compound of formula (I), or a salt thereof, or a prodrug thereof; or a pharmaceutical composition comprising a compound of formula (I), or a salt thereof, or a prodrug thereof is used to treat a drug abuse disorder. For example, a compound of formula (I), or a salt thereof, or a prodrug thereof; or a pharmaceutical composition comprising a compound of formula (I), or a salt thereof, or a prodrug thereof can be used to treat an individual who abuses drugs. For example, the drugs are (including but not limited to) opiates and stimulants. The drugs are specifically (including but not limited to) cocaine, methamphetamine, and marijuana. Alternatively, a compound of formula (I), or a salt thereof, or a prodrug thereof; or a pharmaceutical composition comprising a compound of formula (I), or a salt thereof, or a prodrug thereof can be used to treat an individual who abuses alcohol.

In one embodiment, the compound of formula (I), or a salt thereof or a prodrug thereof; or a pharmaceutical composition comprising the compound of formula (I), or a salt thereof, or a prodrug thereof is used to treat or prevent social dysfunction. The compounds described herein developed by the inventors are used as part of the treatment of psychiatric disorders characterized primarily or secondarily by social dysfunction, such as autism spectrum disorder (ASD), social anxiety disorder (SAD), depression including major depressive disorder (MDD), schizophrenia, and substance use disorders.

Here, the target treatment patient groups include:

Children and adults diagnosed with autism spectrum disorder;

People diagnosed with social anxiety disorder;

Recovering drug (e.g., opiate, stimulant, marijuana) and alcohol addicts seeking to maintain sustained abstinence;

Patients diagnosed with chronic schizophrenia, where the drug can be used as an adjunct to antipsychotic medication to improve social functioning; or

Others who seek to overcome deficits in social functioning as part of their current medical condition (e.g., MDD).

In one embodiment, the condition being treated or prevented is a social dysfunction. For example, the social dysfunction is selected from:

· Not sociable;

Violence

antisocial disorder; or

Drug addiction.

In one embodiment, the subject suffers from a psychiatric disorder. For example, in another embodiment, the subject suffers from a psychiatric disorder wherein asociality is a primary or secondary feature of the psychiatric disorder.

In one embodiment, the subject suffers from a psychiatric disorder selected from the group consisting of:

Autism spectrum disorder;

Social anxiety disorder;

Depression, including major depressive disorder; or

Schizophrenia.

In one embodiment, the subject suffers from or is recovering from a substance abuse disorder, for example, addiction to drugs and chemicals such as cocaine, opium, amphetamines, heroin, alcohol, and nicotine.

In another embodiment, the subject is recovering from a substance abuse disorder and attempting to maintain ongoing abstinence from the drug.

Implementation Method

1. A compound of formula (I), or a salt thereof, or a prodrug thereof:

in:

V is NH, CH ₂ or a direct bond;

W is NH, CH ₂ or a direct bond;

X is NH, CH ₂ or a direct bond;

Y is NH, CH ₂ or a direct bond;

Z is selected from: NH, O, S, S(O), SO ₂ or a direct bond;

R ¹ is selected from H or C(O)R ⁴ ;

R ² is selected from: H, OH, halogen, optionally substituted C _1-5 alkyl or optionally substituted OC _1-5 alkyl;

R ³ is selected from: H, OH, halogen, optionally substituted C _1-5 alkyl or optionally substituted OC _1-5 alkyl;

R ⁴ is an optionally substituted C _1-5 alkyl group;

m is 0 or 1;

n is 0 or 1;

p is 0 or 1; and

q is 0 or 1.

2. The compound of embodiment 1 is of formula (Ia):

in:

Z is selected from: NH, O, S, S(O) or SO ₂ ;

R ¹ is selected from: H or C(O)R ⁴ ;

R ² is selected from: H, halogen, optionally substituted C _1-5 alkyl or optionally substituted OC _1-5 alkyl;

R ³ is selected from: H, halogen, optionally substituted C _1-5 alkyl or optionally substituted OC _1-5 alkyl; and

^R4 is an optionally substituted _C1-5 alkyl group,

or a salt thereof, or a prodrug thereof.

3. The compound of embodiment 1 is of formula (Ib):

in:

R ¹ is selected from H or C(O)R ⁴ ;

R ² is selected from: H, OH, halogen, optionally substituted C _1-5 alkyl or optionally substituted OC _1-5 alkyl;

R ³ is selected from: H, OH, halogen, optionally substituted C 1-5 alkyl or optionally substituted OC _1-5 alkyl; and

^R4 is an optionally substituted _C1-5 alkyl group,

or a salt thereof, or a prodrug thereof.

4. The compound of embodiment 1 is of formula (Ic):

in:

Z is selected from: NH, O, S, S(O) or SO ₂ ;

R ¹ is selected from H or C(O)R ⁴ ;

R ² is selected from: H, OH, halogen, optionally substituted C _1-5 alkyl or optionally substituted OC _1-5 alkyl;

R ³ is selected from the group consisting of: H, OH, halogen, optionally substituted C _1-5 alkyl, or optionally substituted OC _1-5 alkyl; and

^R4 is an optionally substituted _C1-5 alkyl group,

or a salt thereof, or a prodrug thereof.

5. The compound of embodiment 1 is of formula (Id):

in:

Z is selected from: NH, O, S, S(O) or SO ₂ ;

R ¹ is selected from H or C(O)R ⁴ ;

R ² is selected from: H, OH, halogen, optionally substituted C _1-5 alkyl or optionally substituted OC _1-5 alkyl;

R ³ is selected from the group consisting of: H, OH, halogen, optionally substituted C _1-5 alkyl, or optionally substituted OC _1-5 alkyl; and

^R4 is an optionally substituted _C1-5 alkyl group,

or a salt thereof, or a prodrug thereof.

6. The compound of embodiment 1 is of formula (Ie):

in:

Z is selected from: NH, O, S, S(O) or SO ₂ ;

R ¹ is selected from H or C(O)R ⁴ ;

R ² is selected from: H, OH, halogen, optionally substituted C _1-5 alkyl or optionally substituted OC _1-5 alkyl;

R ³ is selected from the group consisting of: H, OH, halogen, optionally substituted C _1-5 alkyl, or optionally substituted OC _1-5 alkyl; and

^R4 is an optionally substituted _C1-5 alkyl group,

or a salt thereof, or a prodrug thereof.

7. The compound of embodiment 1 is of formula (If):

in:

Z is selected from: NH, O, S, S(O) or SO ₂ ;

R ¹ is selected from H or C(O)R ⁴ ;

R ² is selected from: H, OH, halogen, optionally substituted C _1-5 alkyl or optionally substituted OC _1-5 alkyl;

R ³ is selected from the group consisting of: H, OH, halogen, optionally substituted C _1-5 alkyl, or optionally substituted OC _1-5 alkyl; and

^R4 is an optionally substituted _OC1-5 alkyl group,

or a salt thereof, or a prodrug thereof.

8. The compound or salt or prodrug thereof according to any one of embodiments 1 to 7, wherein Z is NH.

9. The compound according to any one of embodiments 1 to 7, or a salt thereof, or a prodrug thereof, wherein Z is O.

10. The compound according to any one of embodiments 1 to 7, or a salt thereof, or a prodrug thereof, wherein Z is S.

11. The compound according to any one of embodiments 1 to 7, or a salt thereof, or a prodrug thereof, wherein Z is S(O).

12. The compound according to any one of Embodiments 1 to 7, or a salt thereof, or a prodrug thereof, wherein Z is SO ₂ .

13. The compound according to any one of embodiments 1 to 12, or a salt thereof, or a prodrug thereof, wherein R ¹ is hydrogen.

14. The compound according to any one of Embodiments 1 to 12, or a salt thereof, or a prodrug thereof, wherein R ¹ is C(O)R ⁴ .

15. A compound as described in any one of embodiments 1 to 12 or 14, or a salt thereof, or a prodrug thereof, wherein ^R4 is an optionally substituted _C1-5 alkyl selected from any one of the following groups: methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, n-pentyl, neopentyl, isopentyl and tert-pentyl.

16. The compound according to any one of embodiments 1 to 12, 14 or 15, or a salt thereof, or a prodrug thereof, wherein R ⁴ is an optionally substituted methyl group.

17. The compound according to any one of embodiments 1 to 16, or a salt thereof, or a prodrug thereof, wherein R ² is hydrogen.

18. The compound according to any one of embodiments 1 to 16, or a salt thereof, or a prodrug thereof, wherein R ² is hydroxyl.

19. The compound according to any one of embodiments 1 to 16, or a salt thereof, or a prodrug thereof, wherein R ² is halogen.

20. The compound according to any one of embodiments 1 to 16 or 19, or a salt thereof, or a prodrug thereof, wherein R ² is fluoro.

21. The compound according to any one of embodiments 1 to 16 or 19, or a salt thereof, or a prodrug thereof, wherein R ² is chloro.

22. The compound according to any one of embodiments 1 to 16, or a salt thereof, or a prodrug thereof, wherein R ² is optionally substituted C _1-5 alkyl.

23. The compound according to any one of embodiments 1 to 16 or 22, or a salt thereof, or a prodrug thereof, wherein R ² is an optionally substituted methyl group.

24. The compound according to any one of embodiments 1 to 16, or a salt thereof, or a prodrug thereof, wherein R ² is optionally substituted OC _1-5 alkyl.

25. The compound according to any one of embodiments 1 to 16 or 24, or a salt thereof, or a prodrug thereof, wherein R ² is optionally substituted methoxy.

26. The compound of any one of example embodiments 1 to 25, or a salt thereof, or a prodrug thereof, wherein R ³ is hydrogen.

27. The compound of any one of example embodiments 1 to 25, or a salt thereof, or a prodrug thereof, wherein R ³ is hydroxy.

28. The compound according to any one of embodiments 1 to 25, or a salt thereof, or a prodrug thereof, wherein R ³ is halogen.

29. The compound of any one of example embodiments 1 to 25 or 28, or a salt thereof, or a prodrug thereof, wherein R ³ is fluoro.

30. The compound according to any one of embodiments 1 to 25 or 28, or a salt thereof, or a prodrug thereof, wherein R ³ is chloro.

31. The compound according to any one of embodiments 1 to 25, or a salt thereof, or a prodrug thereof, wherein R ³ is optionally substituted C _1-5 alkyl.

32. The compound according to any one of embodiments 1 to 25 or 31, or a salt thereof, or a prodrug thereof, wherein R ³ is optionally substituted methyl.

33. The compound according to any one of embodiments 1 to 25, or a salt thereof, or a prodrug thereof, wherein R ³ is optionally substituted OC _1-5 alkyl.

34. The compound according to any one of embodiments 1 to 25 or 33, or a salt thereof, or a prodrug thereof, wherein R ³ is optionally substituted methoxy.

35. The compound according to any one of embodiments 1 to 34, or a salt thereof, or a prodrug thereof, wherein the compound is selected from any one of the following groups:

or a salt thereof, or a prodrug thereof.

36. The compound according to any one of embodiments 1 to 35, or a salt thereof, or a prodrug thereof, wherein the compound is:

or a salt thereof, or a prodrug thereof.

37. The compound according to any one of embodiments 1 to 36, or a salt thereof, or a prodrug thereof, wherein the compound is:

or a salt thereof, or a prodrug thereof.

38. The compound according to any one of embodiments 1 to 36, or a salt thereof, or a prodrug thereof, wherein the compound is:

or a salt thereof, or a prodrug thereof.

39. The compound according to any one of embodiments 1 to 36, or a salt thereof, or a prodrug thereof, wherein the compound is selected from any one of the following groups:

or a salt thereof, or a prodrug thereof.

40. The compound according to any one of embodiments 1 to 39, or a salt thereof, or a prodrug thereof, wherein the compound is a salt of any one of the following compounds: Formula (Ia) or Formula (Ib) or Formula (Ic) or Formula (Id) or Formula (Ie) or Formula (If).

41. The compound according to embodiment 40, or a salt thereof, or a prodrug thereof, wherein the compound is a salt of a compound of formula (Ia).

42. The compound according to embodiment 40, or a salt thereof, or a prodrug thereof, wherein the compound is a salt of a compound of formula (Ib).

43. The compound according to embodiment 40, or a salt thereof, or a prodrug thereof, wherein the compound is a salt of a compound of formula (Ic).

44. The compound according to embodiment 40, or a salt thereof, or a prodrug thereof, wherein the compound is a salt of the compound of formula (Id).

45. The compound according to embodiment 40, or a salt thereof, or a prodrug thereof, wherein the compound is a salt of a compound of formula (Ie).

46. The compound as described in embodiment 40, or a salt thereof, or a prodrug thereof, wherein the compound is a salt of a compound of formula (If).

47. A compound as described in any one of embodiments 1 to 46, or a salt thereof, or a prodrug thereof, wherein the compound is a salt selected from any one of the following: hydrochloric acid, sulfuric acid, phosphoric acid, nitric acid, carbonic acid, boric acid, sulfamic acid, hydrobromic acid, acetic acid, propionic acid, butyric acid, tartaric acid, maleic acid, hydroxymaleic acid, fumaric acid, isethionic acid, malic acid, citric acid, lactic acid, mucic acid, gluconic acid, benzoic acid, succinic acid, oxalic acid, phenylacetic acid, methanesulfonic acid, toluenesulfonic acid, benzenesulfonic acid, salicylic acid, sulfanilic acid, aspartic acid, stearic acid, palmitic acid, oleic acid, lauric acid, pantothenic acid, tannic acid, ascorbic acid and valerate; or a metal salt, including sodium, potassium, lithium, calcium, magnesium and zinc metal salts; or ammonium, alkylammonium salts; or an amino acid salt; or a metal salt, including sodium, potassium, lithium, calcium, magnesium and zinc metal salts; or ammonium, alkylammonium salts; or an amino acid salt.

48. The compound according to any one of embodiments 1 to 47, or a salt thereof, or a prodrug thereof, wherein the compound is a hydrochloride salt.

49. A compound according to any one of embodiments 1 to 48. Or a salt thereof, or a prodrug thereof, wherein the salt is a hydrochloride salt of a compound selected from any one of the following groups:

50. The compound according to any one of embodiments 1 to 49, or a salt thereof, or a prodrug thereof, wherein the salt is a hydrochloride salt of a compound selected from the group consisting of:

51. The compound according to any one of embodiments 1 to 50, or a salt thereof, or a prodrug thereof, wherein the salt is a hydrochloride salt of a compound selected from the group consisting of:

52. The compound according to any one of embodiments 1 to 50, or a salt thereof, or a prodrug thereof, wherein the salt is the hydrochloride salt of:

53. A compound according to any one of embodiments 1 to 48, or a salt thereof, or a prodrug thereof, wherein the compound is a hydrochloride salt of a compound selected from any one of the following groups:

54. A compound as described in any one of embodiments 1 to 39, or a salt thereof, or a prodrug thereof, wherein the compound is a prodrug of a compound described by Formula (Ia) or Formula (Ib) or Formula (Ic) or Formula (Id) or Formula (Ie) or Formula (If).

55. The compound or salt or prodrug thereof according to embodiment 54, wherein the compound is a prodrug of the compound of formula (Ia).

56. The compound or salt or prodrug thereof as described in embodiment 54, wherein the compound is a prodrug of the compound of formula (Ib).

57. The compound or salt or prodrug thereof according to embodiment 54, wherein the compound is a prodrug of the compound of formula (Ic).

58. The compound or salt or prodrug thereof according to embodiment 54, wherein the compound is a prodrug of the compound of formula (Id).

59. The compound or salt or prodrug thereof according to embodiment 54, wherein the compound is a prodrug of the compound of formula (Ie).

60. The compound or salt or prodrug thereof according to embodiment 54, wherein the compound is a prodrug of the compound of formula (If).

61. A compound as described in any one of embodiments 1 to 60, or a salt thereof, or a prodrug thereof, when used as a medicament.

62. A pharmaceutical composition comprising a pharmaceutically acceptable compound according to any one of embodiments 1 to 60, or a pharmaceutically acceptable salt thereof, or a prodrug thereof, and a pharmaceutically acceptable carrier, diluent or excipient.

63. A pharmaceutical composition as described in embodiment 62, wherein the pharmaceutically acceptable compound, or a pharmaceutically acceptable salt thereof, or a prodrug thereof is a compound of formula (Ia), or a salt thereof, or a prodrug thereof.

64. A pharmaceutical composition as described in embodiment 62, wherein the pharmaceutically acceptable compound, or a pharmaceutically acceptable salt thereof, or a prodrug thereof is a compound of formula (Ib), or a salt thereof, or a prodrug thereof.

65. A pharmaceutical composition as described in embodiment 62, wherein the pharmaceutically acceptable compound, or a pharmaceutically acceptable salt thereof, or a prodrug thereof is a compound of formula (Ic), or a salt thereof, or a prodrug thereof.

66. A pharmaceutical composition as described in embodiment 62, wherein the pharmaceutically acceptable compound, or a pharmaceutically acceptable salt thereof, or a prodrug thereof is a compound of formula (Id), or a salt thereof, or a prodrug thereof.

67. A pharmaceutical composition as described in embodiment 62, wherein the pharmaceutically acceptable compound, or a pharmaceutically acceptable salt thereof, or a prodrug thereof is a compound of formula (Ie), or a salt thereof, or a prodrug thereof.

68. A pharmaceutical composition as described in embodiment 62, wherein the pharmaceutically acceptable compound, or a pharmaceutically acceptable salt thereof, or a prodrug thereof is a compound of formula (If), or a salt thereof, or a prodrug thereof.

69. A method for treating or preventing a condition in a subject, the method comprising the following steps: administering to the subject a pharmaceutically acceptable compound as described in any one of embodiments 1 to 60, or a pharmaceutically acceptable salt thereof, or a prodrug thereof; or a pharmaceutical composition as described in any one of embodiments 62 to 68.

70. The method of embodiment 69, wherein the pharmaceutically acceptable compound, or a pharmaceutically acceptable salt thereof, or a prodrug thereof is a compound of formula (Ia), or a salt thereof, or a prodrug thereof.

71. The method of embodiment 69, wherein the pharmaceutically acceptable compound, or a pharmaceutically acceptable salt thereof, or a prodrug thereof is a compound of formula (Ib), or a salt thereof, or a prodrug thereof.

72. The method of embodiment 69, wherein the pharmaceutically acceptable compound, or a pharmaceutically acceptable salt thereof, or a prodrug thereof is a compound of formula (Ic), or a salt thereof, or a prodrug thereof.

73. The method of embodiment 69, wherein the pharmaceutically acceptable compound, or a pharmaceutically acceptable salt thereof, or a prodrug thereof is a compound of formula (Id), or a salt thereof, or a prodrug thereof.

74. The method of embodiment 69, wherein the pharmaceutically acceptable compound, or a pharmaceutically acceptable salt thereof, or a prodrug thereof is a compound of formula (Ie), or a salt thereof, or a prodrug thereof.

75. The method of embodiment 69, wherein the pharmaceutically acceptable compound, or a pharmaceutically acceptable salt thereof, or a prodrug thereof is a compound of formula (If), or a salt thereof, or a prodrug thereof.

76. The method of any one of embodiments 69 to 75, wherein the condition is social dysfunction.

77. The method of any one of embodiments 69 to 76, wherein the condition is a social dysfunction selected from the group consisting of:

· not fitting in; or

Violence; or

antisocial disorder; or

Drug addiction.

78. The method of any one of embodiments 69 to 77, wherein the subject suffers from a mental disorder.

79. The method of embodiment 78, wherein asociality is a primary or secondary feature of the mental disorder.

80. The method of any one of embodiments 69 to 79, wherein the subject suffers from a psychiatric disorder selected from the group consisting of:

Autism spectrum disorder;

Social anxiety disorder;

Depression, including major depressive disorder; or

Schizophrenia.

81. The method of any one of embodiments 69 to 80, wherein the subject suffers from or is recovering from a substance abuse disorder.

82. The method of any one of embodiments 69 to 81, wherein the subject is recovering from a substance abuse disorder and attempting to maintain ongoing abstinence from the drug.

83. Use of a pharmaceutically acceptable compound according to any one of embodiments 1 to 60, or a pharmaceutically acceptable salt thereof, or a prodrug thereof; or a pharmaceutical composition according to any one of embodiments 62 to 68 for treating or preventing a condition in a subject.

84. The use according to embodiment 83, wherein the pharmaceutically acceptable compound, or a pharmaceutically acceptable salt thereof, or a prodrug thereof is a compound of formula (Ia), or a salt thereof, or a prodrug thereof.

85. The use according to embodiment 83, wherein the pharmaceutically acceptable compound, or a pharmaceutically acceptable salt thereof, or a prodrug thereof is a compound of formula (Ib), or a salt thereof, or a prodrug thereof.

86. The use according to embodiment 83, wherein the pharmaceutically acceptable compound, or a pharmaceutically acceptable salt thereof, or a prodrug thereof is a compound of formula (Ic), or a salt thereof, or a prodrug thereof.

87. The use according to embodiment 83, wherein the pharmaceutically acceptable compound, or a pharmaceutically acceptable salt thereof, or a prodrug thereof is a compound of formula (Id), or a salt thereof, or a prodrug thereof.

88. The use according to embodiment 83, wherein the pharmaceutically acceptable compound, or a pharmaceutically acceptable salt thereof, or a prodrug thereof is a compound of formula (Ie), or a salt thereof, or a prodrug thereof.

89. The use according to embodiment 83, wherein the pharmaceutically acceptable compound, or a pharmaceutically acceptable salt thereof, or a prodrug thereof is a compound of formula (If), or a salt thereof, or a prodrug thereof.

90. The use according to any one of embodiments 83 to 89, wherein the disorder is social dysfunction.

91. The use according to any one of embodiments 83 to 90, wherein the disease is a social dysfunction selected from the group consisting of:

· Not sociable;

Violence

antisocial disorder; or

Drug addiction.

92. The use according to any one of embodiments 83 to 91, wherein the subject suffers from a mental disorder.

93. The use according to embodiment 92, wherein asociality is a primary or secondary feature of the mental disorder.

94. The use according to any one of embodiments 83 to 93, wherein the subject suffers from a psychiatric disorder selected from the group consisting of:

Autism spectrum disorder;

Social anxiety disorder;

Depression, including major depressive disorder; or

Schizophrenia.

95. The use of any one of embodiments 83 to 94, wherein the subject suffers from or is recovering from a substance abuse disorder.

96. The use of any one of embodiments 83 to 95, wherein the subject is recovering from a substance abuse disorder and attempting to maintain ongoing abstinence from the drug.

97. The use of a pharmaceutically acceptable compound as described in any one of embodiments 1 to 60, or a pharmaceutically acceptable salt thereof, or a prodrug thereof; or a pharmaceutical composition as described in any one of embodiments 62 to 68, for preparing a medicament for treating or preventing a condition in a subject.

98. The use according to embodiment 97, wherein the pharmaceutically acceptable compound, or a pharmaceutically acceptable salt thereof, or a prodrug thereof is a compound of formula (Ia), or a salt thereof, or a prodrug thereof.

99. The use according to embodiment 97, wherein the pharmaceutically acceptable compound, or a pharmaceutically acceptable salt thereof, or a prodrug thereof is a compound of formula (Ib), or a salt thereof, or a prodrug thereof.

100. The use according to embodiment 97, wherein the pharmaceutically acceptable compound, or a pharmaceutically acceptable salt thereof, or a prodrug thereof is a compound of formula (Ic), or a salt thereof, or a prodrug thereof.

101. The use according to embodiment 97, wherein the pharmaceutically acceptable compound, or a pharmaceutically acceptable salt thereof, or a prodrug thereof is a compound of formula (Id), or a salt thereof, or a prodrug thereof.

102. The use according to embodiment 97, wherein the pharmaceutically acceptable compound, or a pharmaceutically acceptable salt thereof, or a prodrug thereof is a compound of formula (Ie), or a salt thereof, or a prodrug thereof.

103. The use of exemplary embodiment 98, wherein the pharmaceutically acceptable compound or a pharmaceutically acceptable salt or a prodrug thereof is a compound of formula (If) or a salt or a prodrug thereof.

104. The use according to any one of embodiments 98 to 103, wherein the disorder is social dysfunction.

105. The use according to any one of embodiments 98 to 104, wherein the disorder is a social dysfunction selected from the group consisting of:

· Not sociable;

Violence

antisocial disorder; or

Drug addiction.

106. The use according to any one of embodiments 98 to 105, wherein the subject suffers from a mental disorder.

107. The use according to embodiment 106, wherein asociality is a primary or secondary feature of the mental disorder.

108. The use according to any one of embodiments 98 to 107, wherein the subject suffers from a psychiatric disorder selected from the group consisting of:

Autism spectrum disorder;

Social anxiety disorder;

Depression, including major depressive disorder;

Schizophrenia.

109. The use of any one of embodiments 98-108, wherein the subject suffers from or is recovering from a substance abuse disorder.

110. The use of any one of embodiments 98-109, wherein the subject is recovering from a substance abuse disorder and is attempting to maintain ongoing abstinence from the drug.

111. The method of any one of embodiments 69 to 75, wherein the condition is a substance abuse disorder.

112. The method of embodiment 111, wherein the drug is alcohol.

113. The method of embodiment 111, wherein the drug is cocaine.

114. The method of embodiment 111, wherein the drug is methamphetamine.

115. The use of any one of embodiments 83 to 89, wherein the condition is a substance abuse disorder.

116. The use according to embodiment 115, wherein the drug is alcohol.

117. The use according to embodiment 115, wherein the drug is cocaine.

118. The use according to embodiment 115, wherein the drug is methamphetamine.

119. The use according to any one of embodiments 97 to 103, wherein the condition is a substance abuse disorder.

120. The use according to embodiment 119, wherein the drug is alcohol.

121. The use according to embodiment 119, wherein the drug is cocaine.

122. The use according to embodiment 119, wherein the drug is methamphetamine.

Example

The abbreviations used herein are shown in Table 1:

Table 1 Abbreviations used in this manual

abbreviation Full Name/Description APCI atmospheric pressure chemical ionization DCM dichloromethane DMF N,N-dimethylformamide DMSO dimethyl sulfoxide ESI Electrospray ionization EtOH ethanol HRESIMS High-resolution electrospray ionization mass spectrometry HRMS High-resolution mass spectrometry LRMS Low-resolution mass spectrometry MeOH Methanol THF Tetrahydrofuran TLC Thin layer chromatography

General details of the synthesis and characterization methods

Unless otherwise noted, all reactions were carried out under nitrogen or argon atmosphere. THF, toluene and 1,4-dioxane were dried over sodium silk and distilled. In the presence of THF, sodium benzophenone carbonyl, dichloromethane, methanol, ethanol and acetonitrile were distilled from CaH _2. AR grade acetone and commercially available chemicals were used as purchased products. Analytical thin layer chromatography (TLC) was performed using Merck aluminum-based silica gel 60F254 (0.2 mm) plates visualized using short-wave (254 nm) and/or long-wave (365 nm) UV fluorescence. Merck Kieselgel 60 (230-400 mesh) silica gel was used for flash chromatography. Melting points were measured in an open capillary using a Stuart SMP10 melting point instrument, and were uncorrected. Infrared absorption spectra were recorded on a Bruker Alpha FT-IR spectrophotometer, and data were recorded as vibration frequencies (cm ^-1 ). Nuclear magnetic resonance spectra were recorded at 300 K using a Bruker Avance 200 NMR (300.1 MHz) spectrometer. Data are reported as chemical shift (δ ppm), relative integration, multiplicity (s = singlet, br s = broad singlet, d = doublet, dd = doublet, t = triplet, dt = doublet, q = quartet, sep = septet, m = multiplet), coupling constant (J Hz), and assignment relative to residual protonated solvent resonances. Signal assignments were assisted by COSY, DEPT, HSQC, and HMBC experiments, as necessary, relative to residual protonated solvent resonances. Signal assignments were assisted by COSY, DEPT, HSQC, and HMBC experiments, as necessary. LRMS were recorded using electrospray ionization (ESI) or atmospheric pressure chemical ionization (APCI) recorded on a Finnigan LCQ ion trap spectrometer.

Example 1 Synthesis of the compound of formula (I)

Suitable synthetic methods can be found in T.A. Reekie et al., Tetrahedron Letters, 55, 4568–4571, 2014; and T.A. Reekie et al., Synthesis, 45, 3211–3227, 2013.

1-Methyl-1H-pyrazol-5-ol (1)

A magnetically stirred solution of methyl 3-methoxyacrylate (20.0 g, 172.2 mmol) in methanol (100 mL) was slowly treated with N-methylhydrazine (10.0 mL, 189.4 mmol) and heated at reflux for 16 hours. The resulting mixture was cooled to room temperature and concentrated under reduced pressure to afford an orange solid, which was dissolved in N,N-dimethylformamide (DMF) (19.9 mL, 258.4 mmol) and cooled to 0°C. The resulting mixture was slowly treated with phosphorus oxychloride (48.1 mL, 516.6 mmol) and heated at 80°C for 7 hours before being cooled and slowly poured into ice-cold _Na₂CO₃ (600 _mL of a 20% w/v aqueous solution). The aqueous solution was then extracted with chloroform (5 x 100 mL), and the combined organic phases were washed with brine (1 x 100 mL), dried ( _MgSO₄ ), filtered, and concentrated under reduced pressure to afford an orange solid. Recrystallization (dichloromethane/hexane) gave compound 1 (12.87 g, 52%) as yellow crystals, mp 77-78°C (Rf = 0.44 in 1:1 v/v ethyl acetate/hexane).

Table 2 shows the characterization data of Compound 1.

General Process 1

Pre-SOC-2, Pre-SOC-3, Pre-SOC-4, Pre-SOC-5, and Pre-SOC-6 were prepared using General Procedure 1. A magnetically stirred solution of compound 1 (500 mg, 3.46 mmol) in DMF (7 mL) was treated with KOH (387 mg, 6.90 mmol) and the appropriate nitroaniline (3 equivalents, 10.38 mmol), then heated at 100°C for 1 hour. The resulting mixture was cooled to room temperature, treated with _NH₄Cl (100 mL of saturated aqueous solution), and extracted with ethyl acetate (3 x 25 mL). The combined organic phases were washed with brine (1 x 50 mL), dried ( _MgSO₄ ), filtered, and concentrated under reduced pressure to afford a yellow oil. The residue was purified by flash column chromatography (silica gel, 1:4 → 1:1 v/v ethyl acetate/hexane gradient elution), and the relevant fractions were concentrated to afford the desired pyrazole.

General Process 2

FB-SOC-2, FB-SOC-4, FB-SOC-5, and FB-SOC-6 compounds were prepared using General Procedure _2. A suspension of Pre-SOC-2, Pre-SOC-4, Pre-SOC-5, or Pre-SOC-6 (1.20 mmol) and palladium on carbon (10 mg, 10% w/v) in methanol (12 mL) was magnetically stirred for 18 hours at room temperature under H₂ (1 atm). The resulting mixture was filtered through Celite ^™ , and the resulting solid was washed with methanol (3 x 10 mL). The combined filtrates were concentrated under reduced pressure to provide the desired benzodiazepine compound.

1-Methyl-5-((2-nitrophenyl)amino)-1H-pyrazole-4-carbaldehyde (Pre-SOC-1)

A magnetically stirred solution of compound 1 (5.78 g, 40.0 mmol) and 2-nitroaniline (5.52 g, 40.0 mmol) in DMF (40 mL) was treated with powdered KOH (4.49 g, 80.0 mmol) and heated at 120°C for 2 hours. The resulting mixture was cooled to room temperature, treated with _NH4Cl (500 mL of saturated aqueous solution), and extracted with ethyl acetate (3 x 200 mL). The combined organic phases were washed with brine (1 x 100 mL), dried ( _MgSO4 ), filtered, and concentrated under reduced pressure to yield a yellow oil. The residue was purified by flash column chromatography (silica gel, 1:4 → 1:1 v/v ethyl acetate/hexanes gradient elution), and the relevant fractions were concentrated (Rf = 0.33 in 1:1 v/v ethyl acetate/hexanes) to yield compound Pre-SOC-1 (7.38 g, 75%) as a yellow crystalline solid, mp 102-105°C.

Table 2 shows the characterization data of compound Pre-SOC-1.

1-Methyl-1,4,5,10-tetrahydrobenzo[b]pyrazolo[3,4-e][1,4]diazepine (FB-SOC-1)

A suspension of Pre-SOC-1 (3.94 g, 16.0 mmol) and palladium on carbon (100 mg, 10% w/v) in methanol (160 mL) was magnetically stirred at room temperature under _H₂ (1 atm) for 16 hours. The resulting mixture was filtered through Celite ^™ , and the retained solid was washed with methanol (3×20 mL). The combined filtrates were concentrated under reduced pressure to afford FB-SOC-1 (3.04 g, 95%) as a yellow powder, mp 205-207°C (Rf = 0.43 in 1:9 v/v methanol/dichloromethane).

Table 2 shows the characterization data of compound FB-SOC-1.

5-((4-Fluoro-2-nitrophenyl)amino)-1-methyl-1H-pyrazole-4-carbaldehyde (Pre-SOC-2)

Pre-SOC-2 (539 mg, 59%) was prepared according to general procedure 1 from 4-fluoro-2-nitroaniline as an orange solid, m.p. 113-116°C, (Rf = 0.32 in 1:1 v/v ethyl acetate/n-hexane).

Table 2 shows the characterization data of compound Pre-SOC-2.

7-Fluoro-1-methyl-1,4,5,10-tetrahydrobenzo[b]pyrazolo[3,4-e][1,4]diazepine (FB-SOC-2)

Following General Procedure 2, FB-SOC-2 (236 mg, 90%) was prepared as brown crystals, m.p. 181-183°C, (Rf = 0.43 in 1:9 v/v methanol/dichloromethane).

Table 2 shows the characterization data of compound FB-SOC-2.

5-((4-chloro-2-nitrophenyl)amino)-1-methyl-1H-pyrazole-4-carbaldehyde (Pre-SOC-3)

Pre-SOC-3 (563 mg, 58%) was prepared from 4-chloro-2-nitroaniline according to general procedure 1 as yellow crystals. m.p. 131-132°C (Rf = 0.40 in 1:1 v/v ethyl acetate/n-hexane)

Table 2 shows the characterization data of compound Pre-SOC-3.

7-Chloro-1-methyl-1,4,5,10-tetrahydrobenzo[b]pyrazolo[3,4-e][1,4]diazepine (FB-SOC-3)

A magnetically stirred solution of Pre-SOC-3 (337 mg, 1.20 mmol) in methanol (12 mL) was treated with HCl (2.0 mL of a 6.0 M aqueous solution, 12.0 mmol) and then treated with Zn powder (1.57 g, 24.0 mmol). The resulting mixture was stirred at room temperature for 1 hour and then filtered through Celite ^™ . The retained solid was washed with methanol (2×25 mL) and the combined filtrates were concentrated under reduced pressure to give a yellow solid. Recrystallization (n-hexane/dichloromethane) gave FB-SOC-3 (175 mg, 62%) as a brown solid, mp 174-176° C. (Rf=0.25 in 1:9 v/v methanol/dichloromethane).

Table 2 shows the characterization data of compound FB-SOC-3.

1-Methyl-5-((4-methyl-2-nitrophenyl)amino)-1H-pyrazole-4-carbaldehyde (Pre-SOC-4)

Pre-SOC-4 (621 mg, 69%) prepared according to general procedure 1 from 4-methyl-2-nitroaniline was obtained as an orange solid, m.p. 162-163°C (Rf = 0.40 in 1:1 v/v ethyl acetate/n-hexane).

Table 2 shows the characterization data of compound Pre-SOC-4.

1,7-Dimethyl-1,4,5,10-tetrahydrobenzo[b]pyrazolo[3,4-e][1,4]diazepine (FB-SOC-4)

Following general procedure 2, FB-SOC-4 (247 mg, 96%) was prepared as pale yellow crystals, m.p. 172-173°C, (Rf = 0.50 in 1:9 v/v methanol/dichloromethane).

Table 2 shows the characterization data of compound FB-SOC-4.

1-Methyl-5-((2-methyl-6-nitrophenyl)amino)-1H-pyrazole-4-carbaldehyde (Pre-SOC-5)

Pre-SOC-5 was prepared according to General Procedure 1 (but heated at 100°C for 18 hours). Pre-SOC-5 (396 mg, 44%) was obtained as a yellow solid and a 1:1.5 mixture of atropisomers, m.p. 129-130°C (Rf = 0.25 in 1:1 v/v ethyl acetate/n-hexane).

Table 2 shows the characterization data of compound Pre-SOC-5.

1,9-Dimethyl-1,4,5,10-tetrahydrobenzo[b]pyrazolo[3,4-e][1,4]diazepine (FB-SOC-5)

FB-SOC-5 (211 mg, 82%) prepared according to general procedure 2 was obtained as pale yellow crystals, m.p. 137-139°C, (Rf = 0.50 in 1:9 v/v methanol/dichloromethane).

Table 2 shows the characterization data of compound FB-SOC-5.

5-((4-Methoxy-2-nitrophenyl)amino)-1-methyl-1H-pyrazole-4-carbaldehyde (Pre-SOC-6)

Pre-SOC6 (401 mg, 42%) prepared according to general procedure 1 from 4-methoxy-2-nitroaniline was obtained as an orange solid, m.p. 132-133°C (Rf = 0.27 in 1:1 v/v ethyl acetate/n-hexane).

Table 2 shows the characterization data of compound Pre-SOC-6.

7-Methoxy-1-methyl-1,4,5,10-tetrahydrobenzo[b]pyrazolo[3,4-e][1,4]diazepine (FB-SOC-6)

FB-SOC-6 (251 mg, 91%) was prepared according to General Procedure 2 as a pale pink solid, m.p. 126-128°C, (Rf = 0.55 in 1:9 v/v methanol/dichloromethane).

Table 2 shows the characterization data of compound FB-SOC-6.

1-(1-Methyl-4,10-dihydrobenzo[b]pyrazolo[3,4-e][1,4]diazepin-5(1H)-yl)ethan-1-one (FB-SOC-7)

An ice-cold suspension of FB-SOC-1 (200 mg, 1 mmol) in anhydrous DCM (10 mL) and triethylamine (279 μL, 2 mmol) was treated with acetyl chloride (85 μL, 1.2 mmol) and stirred at room temperature for 4 hours. The resulting mixture was diluted with DCM (50 mL) and washed with saturated bicarbonate solution (2×100 mL). The organic phase was separated and washed with brine (50 mL), dried (MgSO ₄ ), and concentrated under reduced pressure to give a yellow powder. Purification by column chromatography (silica gel, eluent: 19:1 to 9:1 v/v DCM:MeOH gradient elution) and concentration of the relevant fractions (Rf=0.40 in 9:1 v/v DCM:MeOH) afforded compound FB-SOC-7 (197 mg, 0.81 mmol, 81%) as a white powder.

1-Methyl-4,5-dihydro-1H-benzo[b]pyrazolo[4,3-f][1,4]oxazepine (FB-SOC-8)

FB-SOC-8 can be synthesized via two multi-step processes (Synthesis 1 and Synthesis 2, respectively).

Synthesis 1

A solution of compound 1 (500 mg, 3.46 mmol), 2-nitrophenol (1.44 g, 10.38 mmol), and powdered KOH (388 mg, 6.92 mmol) in DMF (4 mL) was heated at 120°C in a CEM Explorer™ microwave reactor for 0.5 h. The resulting mixture was cooled to room temperature, treated with _NH₄Cl (50 mL of saturated aqueous solution), and extracted with ethyl acetate (3 x 20 mL). The combined organic phases were washed with brine (1 x 10 mL), dried ( _MgSO₄ ), filtered, and concentrated under reduced pressure to yield a yellow oil. The residue was purified by flash column chromatography (silica gel, 1:4 → 1:1 v/v ethyl acetate/hexane gradient elution), and the relevant fractions were concentrated (Rf = 0.26 in 1:1 v/v ethyl acetate/hexane) to yield 1-methyl-5-(2-nitrophenoxy) _-1H -pyrazole-4-carbaldehyde.

A suspension of 1-methyl-5-(2-nitrophenoxy)-1H-pyrazole-4-carbaldehyde (300 mg, 1.21 mmol) and palladium on carbon (20 mg, 10% w/w) in methanol ( ₁₅ mL) was then magnetically stirred at room temperature under H₂ (1 atm) for 18 hours. The resulting mixture was filtered through Celite ^™ , and the remaining solid was washed with methanol (3 x 20 mL). The combined filtrates were concentrated under reduced pressure to afford a white solid. Recrystallization (dichloromethane/hexane) afforded FB-SOC-8 (229 mg, 94%) as white crystals, mp 143-145°C (Rf = 0.50 in 1:9 v/v methanol/dichloromethane).

Synthesis 2

A magnetically stirred solution of compound 1 (500 mg, 3.46 mmol) and 2-aminophenol (1.13 g, 10.38 mmol) in DMF (4 mL) was treated with powdered KOH (388 mg, 6.92 mmol) and heated at 120°C for 2 hours. The resulting mixture was cooled to room temperature, treated with _NH4Cl (50 mL of a saturated aqueous solution), and extracted with ethyl acetate (3 x 20 mL). The combined organic phases were washed with brine (1 x 10 mL), dried ( _MgSO4 ), filtered, and concentrated under reduced pressure to yield a yellow oil. The residue was purified by flash column chromatography (silica gel, 1:4→1:1 v/v ethyl acetate/hexane gradient elution) and the relevant fractions were concentrated (Rf=0.21 in 1:1 v/v ethyl acetate/hexane) to afford 1-methyl-1H-benzo[b]pyrazolo[4,3-f][1,4]oxazepine (400 mg, 58%) as a brown oil.

1-Methyl-1H-benzo[b]pyrazolo[4,3-f][1,4]oxazepine (350 mg, 1.76 mmol) in methanol (8.5 mL) was then treated portionwise with _NaBH₄ (136 mg, 3.51 mmol) at 0°C for >5 minutes. The resulting mixture was allowed to warm to room temperature and stirred for 1.5 hours before being carefully treated with _H₂O (5 mL). Once hydrogen evolution ceased, the methanol was removed under reduced pressure, and the resulting aqueous mixture was extracted with ethyl acetate (3 x 10 mL). The combined organic phases were dried ( _MgSO₄ ), filtered, and concentrated under reduced pressure to provide FB-SOC-8 as a white powder.

Table 2 shows the characterization data of compound FB-SOC-8.

1-Methyl-4,5-dihydro-1H-benzo[b]pyrazolo[4,3-f][1,4]thiazepine (FB-SOC-9)

A magnetically stirred solution of compound 1 (150 mg, 1.0 mmol), 2-aminothiophenol (120 μL, 1.1 mmol), and piperidine (11 μL, 0.1 mmol) in EtOH (5 mL) was heated at reflux for 3 hours. TLC indicated the absence of starting aldehyde 1. The reaction mixture was cooled to 0°C and treated with _NaBH₄ (98 mg, 2.6 mmol, 2.5 equiv) in portions. Once fully dissolved, the mixture was heated at reflux for an additional hour and then distilled to yield a yellow residue. The yellow residue was dissolved in saturated _NaHCO₃ solution (20 mL) and extracted with ether (3 x 20 mL). The combined organic layers were dried ( _MgSO₄ ), filtered, and concentrated under reduced pressure to yield a yellow solid. The residue was purified by flash column chromatography (silica gel, 3:7→8:2 v/v ethyl acetate/hexane gradient elution) and the relevant fractions were concentrated (Rf=0.15 in 1:1 v/v ethyl acetate/hexane) to give FB-SOC-9 (54 mg, 24%) as a white solid, mp 134-137°C.

Table 2 shows the characterization data of compound FB-SOC-9.

1-Methyl-4,5-dihydro-1H-benzo[b]pyrazolo[4,3-f][1,4]thiazepine-10-oxide (FB-SOC-10)

Compound FB-SOC-10 was synthesized in a multi-step reaction. First, a magnetically stirred solution of SOC-9 (200 mg, 920 μmol) and triethylamine (96 μL, 2.8 mmol) in THF (3 mL) at 0°C was treated dropwise with trifluoroacetic anhydride (200 μL, 1.4 mmol). The resulting solution was stirred at 0°C for 3 hours and then at room temperature for an additional 18 hours. The resulting mixture was then concentrated under reduced pressure to give a yellow oil, which was subjected to flash column chromatography (silica gel, 1:7 v/v ethyl acetate/hexane elution). Concentration of the relevant fractions (Rf = 0.43 in 1:1 v/v ethyl acetate/hexanes) afforded 2,2,2-trifluoro-1-(1-methyl-1H-benzo[b]pyrazolo[4,3-f][1,4]thiazepin-5(4H)-yl)ethanone (261 mg, 94%) as a yellow solid, m.p. 115-118°C.

To prepare FB-SOC-10, a solution of 2,2,2-trifluoro-1-(1-methyl-1H-benzo[b]pyrazolo[4,3-f][1,4]thiazepin-5(4H)-yl)ethanone (90 mg, 273 μmol) and _K₂CO₃ (100 mg, 720 μmol) in _MeOH (6 mL) was magnetically stirred at room temperature for 18 hours. The resulting mixture was then concentrated under reduced pressure, followed by the addition of water (15 mL) and extraction with ethyl acetate (3×15 mL). The combined organic phases were dried ( _MgSO₄ ), filtered, and concentrated under reduced pressure to afford FB-SOC-10 (60 mg, 94%) as a white solid. mp 192-194° C. (Rf=0.45 in 1:9 v/v methanol/dichloromethane).

Table 2 shows the characterization data of compound FB-SOC-10.

1-Methyl-4,5-dihydro-1H-benzo[b]pyrazolo[4,3-f][1,4]thiazepine 10,10-dioxide (FB-SOC-11)

Compound FB-SOC-11 was synthesized in a multistep reaction. First, a magnetically stirred solution of 2,2,2-trifluoro-1-(1-methyl-1H-benzo[b]pyrazolo[4,3-f][1,4]thiazepin-5(4H)-yl)ethanone, prepared as described for the synthesis of SOC-10 (50 mg, 160 μmol), in glacial acetic acid (0.6 mL) was treated dropwise with hydrogen peroxide (50 μL of a 30% aqueous solution, 480 μmol) at room temperature and then heated to 80°C for 4 hours. The resulting solution was cooled to room temperature, treated with water (15 mL), and extracted with ethyl acetate (3 × 15 mL). The combined organic phases were dried (MgSO ₄ ), filtered and concentrated under reduced pressure to afford 2,2,2-trifluoro-1-(1-methyl-10,10-dioxo-1H-benzo[b]pyrazolo[4,3f][1,4]thiazepin-5(4H)-yl)ethanone (55 mg, 80%) as a yellow solid, mp 187-190° C. (Rf=0.25 in 1:1 v/v ethyl acetate/hexanes).

A solution of 2,2,2-trifluoro-1-(1-methyl-10,10-dioxo-1H-benzo[b]pyrazolo[4,3-f][1,4]thiazepin-5(4H)-yl)ethanone (56 mg, 159 μmol) and K ₂ CO ₃ (62 mg, 450 μmol) in MeOH (4 mL) was then magnetically stirred at room temperature for 18 hours. The resulting mixture was then concentrated under reduced pressure, followed by the addition of water (15 mL) and extraction with ethyl acetate (3×15 mL). The combined organic phases were dried (MgSO ₄ ), filtered, and concentrated under reduced pressure to afford FB-SOC-11 (39 mg, 97%) as a white solid. (Rf=0.39 in 1:9 v/v methanol/dichloromethane).

Table 2 shows the characterization data of compound FB-SOC-11.

Table 2 - Characterization data of selected compounds

Example 2 Salt

The preparation of salts is known to those skilled in the art. For example, to prepare the hydrochloride salt, HCl (4M in dioxane, 1.1 equivalents) can be added to a magnetically stirred solution of the free base, such as (FB-SOC-1 or FB-SOC-2) in THF. Stirring is continued for 30 minutes, and the resulting precipitate is collected by filtration to yield the corresponding salt derivative, such as the hydrochloride salt.

An exemplary salt of formula (I) is the hydrochloride salt of:

hydrochloride

It is used in the examples below and is given the code SOC-1.

Example 3 - In vivo study (Behavioral effects - social interaction test in Hooded Wistar rats)

A social interaction test using hooded Wistar rats showed that SOC-1 elicited a powerful prosocial effect in rodents. The primary phenotypic effect was a significant increase in a behavior known as "adjacent lying," in which unfamiliar rats, which have been in passive social contact for extended periods, gather together. As shown in Figure 1, image (A), intraperitoneal (IP) administration of 5 and 10 mg/kg SOC-1 to male hooded Wistar rats elicited a significant increase in adjacent lying in the social interaction test (n=6-8 pairs per condition, 20-minute test). Notably, the effects of SOC-1 extended beyond its acute administration, with a sustained upregulation of social behavior (measured by time spent in proximity) observed in rats given only three doses of SOC-1 two weeks prior (Figure 1, image B). After two weeks of SOC-1 treatment (three doses total over six days), male hooded Wistar rats showed an increase in proximity to a novel conspecific relative to controls (time spent within one body length). Similar long-term upregulation of social interaction was found with oxytocin (OT) (M. Bowen et al., "Adolescent Oxytocin Exposure Causes Persistent Reductions in Anxiety and Alcohol Consumption and Enhances Sociability in Rats", PloS One, 6(11), 2011-e27237; and A. Suraev et al., "Adolescent exposure to oxytocin, but not the selective oxytocin receptor agonist TGOT, increases social behavior and plasma oxytocin in adulthood", Hormones and Behavior, 65(5), 488-496, 2014.). Direct comparison of the most effective dose of SOC-1 with other drugs known to induce neighborly lying behavior in rats (arginine vasopressin (AVP), 3,4-methylenedioxy-methamphetamine (MDMA), and WAY 267,464) showed that SOC-1 had the most powerful prosocial effect (Figure 1, image (C)).

Administration of SOC-1 to male Albino Wistar rats showed an increase in social preference when choosing between spending time with a caged conspecific or a toy rat (Figure 2, Image (A)). As shown in Figure 2, Image (A), a dose of 5 mg/kg of SOC-1 significantly increased preference for live rats.

Example 4 - In vivo study (behavioral effects - non-human addiction using macaques)

Additional behavioral studies using SOC-1 in nonhuman primate models of addiction and social interaction showed that oral administration of SOC-1 to rhesus monkeys at a dose of 3 mg/kg nearly halved intravenous cocaine self-administration relative to baseline levels without affecting food consumption (Figure 2, image (B)). In this study, administration of SOC-1 significantly reduced cocaine consumption without affecting food consumption. Figure 3 shows the cumulative proportion of cocaine self-administration.

Example 5 - In vivo studies (pharmacokinetics)

In an in vivo rodent model using hooked Wistar rats, SOC-1 exhibited excellent oral bioavailability and pharmacokinetic parameters relative to the behaviorally effective intraperitoneal route of administration (Table 3 and Figure 4). When administered by intraperitoneal injection (IP), SOC-1 plasma levels showed a rapid rise. They declined rapidly within the first hour, and then the concentration decreased throughout the test time. In contrast, oral administration of the same dose of SOC-1 showed higher plasma concentration stability over many hours. When SOC-1 was administered orally, the initial increase in plasma levels was not obvious, but the rate of decline was smaller, and the plasma concentration was equivalent to the concentration between 1 and 4 hours in the IP administration method. In many ways, this is a more favorable situation. Unlike IP administration, oral administration results in SOC-1 levels that remain high but are also very stable over many hours, with a half-life of more than four hours.

Table 3 - Pharmacokinetic data of intraperitoneal (IP) and oral (PO) administration of SOC-1

SOC-1 demonstrated excellent stability in human plasma in an in vitro plasma stability assay (Figure 5). Even at a relatively low concentration (500 nM), SOC-1 exhibited a half-life exceeding 6 hours when incubated with human plasma at human body temperature.

Example 6 - In vivo studies (pharmacodynamics)

Fos immunohistochemistry was used to characterize the neural "signature" of SOC-1 (5 mg/kg) in the brains of rats treated with peripheral oxytocin (OT) injections (1 mg/kg). As shown in Table 4, SOC-1 produced an overall pattern of neural activation and behavioral effects similar to OT. However, these effects were much larger, and SOC-1 activated additional regions implicated in the regulation of social behavior (e.g., the lateral septum, medial preoptic area, and supraoptic nucleus).

SOC-1 produced robust activation of OT-containing neurons in the supraoptic nucleus (Figure 6: (A) = vehicle; (B) = oxytocin; and (C) = SOC-1) and paraventricular nucleus of the hypothalamus. Administration of SOC-1 (5 mg/kg) to male Wistar rats strongly activated Fos expression in oxytocin-positive cells in the supraoptic nucleus (SON) of the hypothalamus. SOC-1 activated these oxytocin-containing cells in the SON more effectively than 1 mg/kg oxytocin (intraperitoneally).

Table 4 - Mean number of Fos-positive cells in brain regions of interest (± standard error of the mean (SEM))

^a) SOC-1 is significantly greater than VEH; ^b) SOC-1 is significantly greater than OT

However, it is noteworthy that SOC-1 does not show affinity for the orthosteric binding site on the oxytocin receptor (OTR) (human OT receptor, IC50 (nM) for [3H]OT=>10,000). In addition, detection of IP1 signaling on OTR HEK cells showed that SOC-1 is not an agonist or antagonist of OTR, nor is it a positive allosteric modulator of OTR. SOC-1 also does not show any affinity for the orthosteric binding site of AVP receptors (V1A, V1B, or V2) (such as the human V1a receptor, IC50 (nM) for [3H]AVP=>10,000). IP1 signaling on V1aR HEK cells revealed that SOC-1 is not an agonist or positive allosteric modulator of V1aR, but instead displays weak antagonist activity at these receptors.

Despite the apparent strong OT-like behavioral effects of this compound in vivo and the clear activation of the brain OT system seen in our Fos data, comprehensive screening for affinity of orthosteric binding sites at numerous receptors from the Psychoactive Drug Screening Program (PDSP) has not identified a target. In PDSP, SOC-1 had no significant affinity for 5-HT1A, 5-HT1B, 5-HT1D, 5-HT1e, 5-HT2A, 5-HT2B, 5-HT2C, 5-HT3, 5-ht5a, 5-HT6, 5-HT7, Alpha1B, Alpha1D, Alpha2A, Alpha2B, Alpha2C, Beta1, Beta2, Beta3, D1, D2, D3, D4, D5, DAT, DOR, GABAA, H1, H2, H3, KOR, M1, M2, M3, M4, M5, MOR, NET, PBR, SERT, Sigma 1 and Sigma 2 receptors in BZP rat brain.

Signaling assays using rat hypothalamic neurons showed that SOC-1 did not alter IP1 levels and could not reliably alter Ca2 ⁺ levels. However, SOC-1 was able to reduce basal cAMP but not forskolin-stimulated cAMP. All of this suggests that it may act on Gs-coupled receptors as an inverse agonist or as an agonist for Gs-coupled receptors.

Example 7 Toxicity and adverse reaction testing

SOC-1 has no apparent in vitro or in vivo cytotoxicity. In one assay, SH-SY5Y neuroblastoma cells and HEK-293 cells (4×10 ⁴ cells per well) were exposed to OT or SOC-1 (0.01-10 μM) for 48 hours, with daily medium changes. Cell viability was assessed 4 hours after exposure to cytochrome blue (Promega). There were no significant differences in viability induced by OT or SOC-1 at any of the concentrations tested (Table 5). Since SH-SY5Y cells express OTR (P. Cassoni et al., Int J Cancer, 77, 695-700, 1998) and HEK-293 cells express low amounts of endogenous OTR (BMC Genomics 12:4), these results suggest that SOC-1 does not induce receptor- or non-receptor-mediated cytotoxicity, even at high concentrations.

Table 5 - Cell viability after 48 hours of exposure to oxytocin or SOC-1. Viability is expressed as the mean percentage of vehicle viability ± standard deviation (SD). There were no significant differences in viability induced by oxytocin or SOC-1 at any concentration tested.

Regarding in vivo toxicity/adverse events: At higher doses in rats (10-20 mg/kg IP), there was some mild suppression of locomotor activity. However, these rats were not ataxic or comatose; they were simply in a state of relaxed immobility. Rats tolerated repeated dosing with SOC-1 without any apparent adverse effects, such as weight loss or changes in fur condition. SOC-1 had no effect on conventional non-social anxiety tests (e.g., elevated plus maze, open field), either during acute administration or during residual long-term effects. SOC-1 did not induce conditioned place preference in rats, suggesting a low abuse potential. Biotelemetry experiments revealed that rats (5 mg/kg) maintained a mild hypothermia, but SOC-1 had no effect on heart rate (Figures 7 and 8). Pretreatment with the vasopressin V1A receptor antagonist SR49059 (10 mg/kg) did not block the effects on body temperature and other behaviors. At higher ambient temperatures (i.e., 30°C), the mild hypothermia produced by SOC-1 disappeared, but the drug's social effects were maintained at these temperatures. Data in both studies were collected using the DataSciences International Biotelemetry System.

Example 8 - Drug Relapse Study in Sprague-Dawley Rats

animal

Thirty-two male Sprague Dawley rats (Perth, Western Australia, Australia) (weighing 200–250 g) were obtained from the Animal Resource Center. Rats were housed in pairs (cage size: 40 x 27 x 16 cm) except for 2 days post-surgery, when they were individually housed until 6 weeks after surgery, when they were transferred to larger cages: 64 x 20 x 40 cm. Food and water were available during home confinement except during experimental procedures. Lighting was maintained on a 12-h light/dark cycle (lights on at 06:00), and all experiments were conducted during the light cycle. Room temperature was maintained at 21°C (±1°C). Rats were acclimated to the facility for seven days before the start of the experiment and were handled daily for an additional seven days. All experimental procedures were performed in accordance with the Australian Code of Practice for the Care and Use of Animals for Scientific Purposes (8th edition, 2013) and were approved by the Macquarie University Animal Ethics Committee.

drug

Methamphetamine hydrochloride (METH) was purchased from the Australian Government Analytical Laboratory (Pymble, New South Wales, Australia). SOC-1 was synthesized by Professor Michael Kassiou (School of Chemistry, University of Sydney, Australia). METH and SOC-1 were dissolved in saline (0.9%) for injection. The V1a receptor antagonist SR49059 was purchased from Axon Medchem BV (Groningen, the Netherlands). The oxytocin receptor antagonist C25 (5-(3-(3-(2-chloro-4-fluorophenoxy)azetidin-1-yl)-5-(methoxymethyl)-4H-1,2,4-triazol-4-yl)-2-methoxypyridine) was synthesized according to the methods of Hudsen et al. (2005) and Brown et al. (2010). SR49059 and C25 were dissolved in 15% dimethyl sulfoxide (DMSO), 2% Tween 80, and 83% saline vehicle. All solutions were freshly prepared at each stage.

instrument

The test was performed in 16 standard operating reaction chambers (32-25-34 cm; Med Associates, St. Albans, Vermont, USA), which were placed in a sound-attenuated box (41x56x56 cm) equipped with a fan to block noise and provide ventilation. Each reaction chamber was equipped with two retractable rods (1 active and 1 inactive), with a warning light and a room light located above each rod. The reaction chambers also included a metal arm with adjustable weight and a spring connector connected to a swivel joint. The polyethylene tubing passing through the spring connector was connected to a 10 ml syringe of an infusion pump (Med Associates) located outside the sound attenuation chamber. The tube discharged from the bottom of the spring connector was connected to the back support of the intravenous catheter.

Four infrared beam detectors were also placed on the side walls of each operant chamber to measure locomotor activity. Active and inactive lever pressures, infusion times, and locomotor activity were collected and recorded using MED-PC software.

Operation

Rats were implanted with a chronic indwelling catheter in the right jugular vein. To achieve this, the rats were anesthetized with isoflurane gas (3% oxygen at 2 l/min) and sterile surgical techniques were used. Catheter implantation and catheter construction were as previously described (Moteby et al., 2013). Rats were treated with 0.2 ml of the antibiotic cefazolin sodium (100 mg/ml) intravenously and the analgesic caspofungin (5 mg/kg) subcutaneously at the time of surgery and daily for the next two days. Thereafter, catheter patency was maintained by daily intravenous flushing with 0.2 ml of cefazolin sodium in heparinized saline (300 IU/ml). Rats were allowed to recover from surgery for 7 days before the start of the experiment.

Methamphetamine intravenous self-administration (IVSA) process

After recovery from surgery, rats were able to self-administer METH intravenously over a 2-hour period. At the beginning of each phase, the catheter was flushed with 0.1 ml of heparinized saline (10 IU/ml) and the infusion line was connected. The start of the phase was indicated by rod extension and room lighting. The rods were assigned as active or inactive, and the position of the active rod was balanced throughout the chambers. Depression of the active rod delivered a 3-second infusion of METH (0.1 mg/kg/infusion, 50 μL), illuminated the cue light, and extinguished the house lights. The lights in the room were turned off for 20 seconds, during which time depression of the active rod had no procedural consequences. Depression of the inactive rod had no procedural consequences at any time. At the end of each treatment, the infusion line was disconnected and the catheter was flushed with 0.2 ml of cefazolin sodium in heparinized saline.

Before starting the IVSA procedure, rats were acclimated to the chamber. The rod was retracted and no fluids could be administered. This phase lasted for 60 minutes.

Experiment 1 - Acquisition and preservation of METH IVSA

Rats were trained to self-administer METH in a 2-hour fixed-ratio, 1-time schedule. To avoid overdose, each rat was limited to 60 infusions per session. Rats were continued on a FR-1 schedule until a stable response was achieved, as indicated by a 10% change in infusion and active lever pressure compared to the previous day. A stable baseline for self-administration was achieved over 10 days, after which the rats entered the progressive ratio phase.

Progressive Ratio Stage

During the progressive ratio (PR) phase, METH infusions were delivered at the following ratio: ((5 x (e ^{boost # x 0.2} ) - 5)). The number of active lever presses required to receive one METH infusion was based on the following sequence: 1, 2, 4, 6, 9, 12, 15, 20, 25, 32, 40, 50, 62, 77, 95, 118, 145, 178, 219, 268 (Cornish et al., 2005); each day's PR phase was completed 2 hours later; or if the infusion was not self-administered for 60 minutes. Rats continued the PR phase until lever pressing was stable, as determined by two consecutive days of infusion and a change in the number of lever presses of less than 10%. Lever pressing stabilized after 7 days. During this period, rats were given an intraperitoneal injection of simulated saline (1 ml/kg) before this phase.

In case the bar is pressed steadily, SOC-1 is started to be tested. Each test day is separated by 2-3 PR predetermined phases to regain the baseline level of METH infusion response. Rats receive SOC-1 in a dose order (0, 1.25, 2.5, 5 and 10 mg/kg) that increases progressively, and it is administered 15 minutes before the test period. During the last test, rats receive vehicle injection, and to ensure that they continue to press the bar similar to that tested for the first time, the bar that is significantly reduced on the SOC-1 test day is pressed because the effect of the compound, rather than injection pressure.

Experiment 2 - Acquisition and preservation of METH IVSA

The acquisition and maintenance of METH IVSA in Experiment 2 were the same as those in Experiment 1, except that the IVSA phase was conducted 5 days per week for 21 days.

eliminate

After the last day of METH self-administration, rats were exposed to a 2-hour daily extinction phase. This phase was identical to the self-administration phase, except that the activation lever press resulted in saline infusion coupled with a reminder light, and the maximum output criterion was omitted. Rats continued under extinction conditions for at least ten days, until lever presses were <25 per phase for two consecutive days. In the second week of extinction, rats were given a simulated saline solution (1 ml/kg saline) intraperitoneally 2 hours prior to injection.

recover

Once the elimination criteria are met, rats undergo a recovery trial. Each recovery trial phase is separated by three elimination days. Prior to each recovery trial phase, rats receive a first injection (DMSO and Tween vehicle, C25 or SR49059) 5 minutes before the second injection (saline vehicle or SOC-1), after which they receive their final injection (saline vehicle or METH) 5 minutes later and are immediately placed in the chamber for 2 hours to measure rod pressing and motor activity.

Rats initially received SOC-1 in an ascending dose order (0, 2.5, 5, 10 mg/kg, intraperitoneal) and then received METH (1 mg/kg, intraperitoneal). After the first four recovery periods were completed, antagonist (C25 at a dose of 10 mg/kg, intraperitoneal or SR49059 at a dose of 1 mg/kg, intraperitoneal) pretreatment was administered before SOC-1 (10 mg/kg) and METH administration. Antagonist injections were balanced between the two recovery periods, so that before each phase, half of the rats received C25 and the other half received SR49059. During the post-antagonism recovery phase, rats received the highest tested dose of SOC-1, accompanied by vehicle injection, to ensure that SOC-1 alone did not change the rod pressing activity. In order to also ensure that rats continued to recover to METH, after completing the SOC-1 ascending dose test phase and the final recovery test phase, an additional recovery test phase was also included in which rats received an intraperitoneal injection of vehicle before the METH trigger drug. Recovery conditions were the same as in the elimination phase.

Statistical analysis

The ratio of active to inactive lever presses per day during the acquisition period was analyzed using a two-way repeated-measures analysis of variance (ANOVA) followed by planned pairwise comparisons. Repeated-measures ANOVAs were also used to compare the number of infusions and active lever presses over a 20-day period to ensure rats acquired methotrexate self-administration, followed by planned pairwise comparisons. Locomotor activity throughout the self-administration period was analyzed using repeated-measures ANOVAs with planned pairwise comparisons.

The effects of SOC-1 on methotrexate (METH) intake and active lever pressing were analyzed using repeated-measures ANOVA followed by planned pairwise comparisons. Comparisons of active and inactive lever pressing after drug administration were performed using a two-way repeated-measures ANOVA. Locomotor activity after SOC-1 administration was also analyzed using repeated-measures ANOVA followed by planned pairwise comparisons.

To assess whether rats had extinguished METH-paired responding, mean active lever presses during the last three METH self-administration sessions were compared with active lever presses during the inactivation period using repeated-measures ANOVA with planned comparisons, as were changes in locomotor activity.

To determine whether rats resumed METH-paired lever responding, active lever presses from the last day of extinction before reinstatement were compared to active lever presses during the reinstatement test using a two-way repeated measures ANOVA. A two-way repeated measures ANOVA was used to compare active and inactive lever presses used during reinstatement to ensure that rats continued to discriminate between levers. Motor behavior during the extinction and reinstatement periods was also analyzed using a two-way repeated measures ANOVA. The effect of SOC-1 on reinstatement of METH-seeking behavior was assessed using a repeated measures ANOVA followed by planned pairwise comparisons. Statistical analyses were performed using SPSS 20 Graduate Edition for Mac. Greenhouse-Geisser corrections were reported in cases where the assumption of sphericity was violated. Statistical significance was set at P < 0.05.

result

Testing SOC-1 in Progressive Ratio Tables

Figures 9, 10, and 11 show the effects of SOC-1 or vehicle on a) infusion (Figure 9), b) lever pressing (Figure 10), and c) locomotor behavior (Figure 11) in rats that self-administered methamphetamine according to a progressive ratio schedule. In these figures, * indicates p<0.05 compared to vehicle.

After recovery from right jugular vein catheter implantation surgery, male Sprague Dawley rats were trained to produce rod pressing for methamphetamine (0.1mg/kg/ infusion) in 2 hours FR-1 scheduled phase every day. Stable reaction was completed in 10 days under the FR-1 schedule. Then rats were transferred to the PR schedule and kept for 7 days until their reaction was stable. Once their rod pressing was stabilized, the SOC-1 test was carried out. Each test day was separated by the 2-3 PR scheduled phase. Rats were administered SOC-1 in the order of increasing dosage (0,1.25, 2.5,5 and 10mg/kg, intraperitoneal), and it was administered in the first 15 minutes of the test phase. During the last test, rats received vehicle injection.

Administration of SOC-1 at doses of 5 mg (p = 0.043) and 10 mg/kg (p = 0.007) significantly reduced lever presses on the PR scale compared to vehicle administration. Infusion breakpoints and locomotor activity were significantly reduced after the 10 mg/kg dose compared to vehicle administration (p = 0.001 and p = 0.005, respectively). There were no significant differences in lever presses (p = 0.236), infusion breakpoints (p = 0.191), or locomotor activity (p = 0.107) compared to the previous vehicle recovery phase.

Testing SOC-1 for methamphetamine-induced reinstatement

Figure 12 and Figure 13 are presented at the recovery period that methamphetamine causes, and vehicle, SOC-1 and SOC-1 are used in combination with C25 or SR49059 to the influence of a) active bar pressing (Figure 12) and b) motor behavior (Figure 13).In these figures, # represents the main effect p<0.05 of day, & represents the main effect p<0.05 of processing, and * represents relative to vehicle+METH condition p<0.05.

After recovery from right jugular vein catheter implantation surgery, male Sprague Dawley rats are allowed to self-administer methamphetamine (0.1mg/kg/infusion) in a 2-hour FR-1 scheduled phase carried out 5 days per week for 21 days. Afterwards, rats are exposed to a 2-hour elimination process every day. Rat reaches elimination criterion (at least 10 days, less than 25 activity bar presses for two consecutive days) after 15 times. Once elimination criterion is met, rats undergo recovery testing. Each recovery testing phase is separated by three elimination days. Before each recovery phase, rats receive first injection (DMSO and Tween vehicle, C25 or SR49059) 5 minutes before the second injection (saline vehicle or SOC-1), after which they receive their final injection (saline vehicle or methamphetamine), are immediately placed in the reaction chamber 2 hours after 5 minutes. Treatment regimen is as follows: rats are initially administered SOC-1 in ascending dose order (0, 2.5, 5, 10mg/kg, intraperitoneal), and subsequently methamphetamine (1mg/kg, intraperitoneal) is administered. Afterwards, before the SOC-1 administration of 10mg/kg intraperitoneal dosage and methamphetamine triggering, antagonist pretreatment (10mg/kg C25, intraperitoneal, or 1mg/kg SR49059, intraperitoneal) is applied. In the next stage, rats receive 10mg/kgSOC-1 before vehicle injection administration, to ensure that SOC-1 is administered alone and does not change the bar pressing activity. In the recovery phase and the final recovery phase after SOC-1 dosage test, other vehicle+methamphetamine treatment is carried out to ensure that rats continue to recover.

Compared to the previous elimination day, rats returned to their previous lever-pressing activity, as indicated by a significant increase in lever-pressing activity (p < 0.005). Comparison of vehicle administration with drug treatment during the recovery period showed that SOC-1 administration at doses of 2.5 mg (p < 0.005), 5 mg (p < 0.005), and 10 mg/kg (p < 0.005) significantly reduced methamphetamine-induced lever-pressing activity. Administration of an oxytocin receptor antagonist (C25) or a Via receptor antagonist (SR49059) failed to block the attenuating effect of SOC-1 administration on methamphetamine-induced reinstatement, as indicated by a significant decrease in lever-pressing activity (p < 0.005). Administration of SOC-1 alone (SOC-1 + vehicle) did not significantly alter lever-pressing activity compared to the day before elimination (p = 0.147). During an additional vehicle-based reinstatement phase, rats continued to return to their previous lever-pressing activity (p < 0.05).

Locomotor activity was also significantly higher during the recovery phase than during the elimination phase (p < 0.005). During the recovery phase, locomotor activity was significantly reduced when 2.5 mg (p = 0.006), 5 mg (p < 0.005), and 10 mg/kg (p < 0.005) of SOC-1 were administered before methamphetamine induction. Similar significant effects were observed when oxytocin receptor (p < 0.005) and Via receptor antagonists (p < 0.005) were administered before SOC-1 and methamphetamine administration. SOC-1 alone did not significantly alter locomotor activity during the recovery phase compared to the preceding elimination phase (p = 0.472).

Example 9 Acute Toxicity Evaluation of a Single Bolus Dose of a SOC-1 Test Item Administered Intravenously to Sprague-Dawley Rats in a One-Week Dose Ranging Study

The acute toxicity of the SOC-1 test item was evaluated in Sprague Dawley rats following a single intravenous bolus dose. Four groups of three adult female rats were treated with 1, 3, 10, and 30 mg/kg SOC-1 and then observed for 7 days without autopsy before termination on study day 8.

In this study, treatment with the SOC-1 test article was well tolerated at all dose levels. After day 1 of this study, dose-related findings of mild piloerection, hunched posture and gait, and eyelid closure were detected. These findings typically occurred within 4-24 hours and were considered treatment-related. There was also a finding of exaggerated flinching and vocalization at the dose that was considered treatment-related.

In this study, the acute tolerated dose of SOC-1 administered as an intravenous bolus was identified to be at least 30 mg/kg.

The testing was conducted in accordance with the Organization for Economic Cooperation and Development (OECD) Principles on Good Laboratory Practice (GLP), revised in 1997. The study, conducted in accordance with OECD and U.S. Food and Drug Administration GLP standards, is accepted under the OECD Mutual Recognition of Data Arrangement (including the U.S. and Japan).

The primary objectives of this study were to investigate the tolerability and acute toxicity of the SOC-1 test item following a single bolus dose administered via the intravenous route to adult Sprague Dawley rats.

This study was conducted in accordance with the guidelines set by the Australian Health and Medical Research Council, Australian Code of Practice for the Care and Use of Animals for Scientific Purposes, 8th edition, 2013 (1). This study was assessed and approved by the University of Queensland Animal Ethics Committee.

Test and vehicle/control items

The "Test Items" and "Vehicle/Control Items" are shown in Tables 6 and 7, respectively.

Table 6 Test items used in this embodiment

Table 7 Carriers/controls used in this example

Test and control formulations

SOC-1 test items were dissolved in 0.9% saline for dosing according to Table 8. Formulations were prepared under clean conditions in a laminar flow cabinet using sterile polypropylene tubes and gently swirled/vortexed to ensure complete dissolution of the compound in the vehicle used as before.

Table 8 - Dosage Formulations Used in This Example

The test item formulations were kept at ambient temperature on each day of use (and used on the same day). No dose formulation samples were collected for this study.

GLP storage

Unused test items are kept in the test equipment under appropriate storage conditions.

animal

Table 9 shows the details regarding the animals used in the study.

Table 9 - Animals used in this example

The test system environment is shown in Table 10.

Table 10 - Test system environment used in this example

method

The design of the acute single-dose toxicity study in rats was adapted from the OECD Guideline for the Testing of Chemicals No. 420, Acute Oral Toxicity – Fixed-Dose Method (2001) (2) and the test facility standard operating procedure SP_T002 “Dose-Ranging Study in Rodents”. The study procedures are described below and were conducted in accordance with the relevant test facility standard operating procedures.

The study design is summarized in Table 11.

Table 11 Study design used in this example

The start of the study was staggered by 2 days between Cohorts 2 and 3 to allow for adjustments at each dose level based on toxicity from the previous dose.

Briefly, after the acclimation period, the study began, with treatments administered on study day 1. Animals were observed for signs of toxicity throughout the study, as described below, and body weights were collected daily. Surviving animals were sacrificed on study day 8 without necropsy.

treat

Test item treatments were administered as an intravenous bolus at a dose volume of 2 mL/kg on day 1 of the study in restrained, conscious animals (except Group 4, where lightly anesthetized animals were treated at a dose of 6 mL/kg after administration of isoflurane anesthesia).

observe

Morbidity and mortality were recorded daily during the acclimation period and on day 8, and twice daily from day 1 to day 7.

Conduct at least one clinical observation during the acclimation period. Clinical observations should be conducted at least once from Day 1 until the day before termination. On the day of treatment, animals should be monitored continuously for the first 30 minutes after treatment, with formal observations conducted 30 minutes, 1 hour, and 4 hours after treatment.

Clinical observations include examining the animal for changes in the skin and coat, eyes and mucous membranes, respiratory and circulatory function, gait and posture, behavior, tremors or seizures, and any other abnormalities.

Clinical observations also included daily inspection of the tail for any injection site reactions (eg, erythema or edema).

weight

Body weights were recorded daily during the acclimation period and during treatment from day 1 to day 8. The mean body weight values for study days 1-8 are shown in FIG14 as group means ± standard deviation (SD).

autopsy

On day 8, all animals underwent final body weight measurement and were then euthanized by an overdose of pentobarbital (intraperitoneally). Carcasses were discarded without autopsy.

Data collection and evaluation

Data were collected using Provantis v.9.3.1.1 or recorded in an appropriate TetraQ format and then tabulated using Provantis v.9.3.1.1, GraphPad Prism 6, and/or Excel 2010.

Morbidity and mortality

There were no unplanned deaths or morbidity events in this study.

Clinical observation

In rats treated with 10 and 30 mg/kg of the SOC-1 test item, piloerection, hunched posture and gait, and eyelid closure were observed on study day 1. These findings were graded as mild in severity, with the exception of one rat with moderate eyelid closure. These findings were generally detected within 30 minutes of treatment and resolved within 4-24 hours after dosing, with the exception of one mild piloerection that resolved by day 3. These findings were considered treatment-related.

Group 3 rats also showed exaggerated flinching and vocalizations at the 10 mg/kg dose. This finding is considered treatment-related. Rats were then given 30 mg/kg of isoflurane anesthesia prior to dosing to facilitate administration and minimize pain/distress.

No injection site reactions were noted.

Clinical observations are summarized in Table 12.

Table 12 - Summary of clinical signs incidence in this example

SOC-1 test item treatment had no effect on body weight; all animals in this study gained weight.

Body weight values are summarized in Figure 14 and Table 13.

Table 13 - Average body weight of rats used in Groups 1 to 4 in this example

in conclusion

In this study, treatment with the SOC-1 test item was tolerable at dose levels of 1, 3, 10, and 30 mg/kg. Dose-related findings of piloerection, hunched posture and gait, and eyelid closure were noted after treatment on study day 1. These findings generally resolved within 4-24 hours and were considered treatment-related. At the time of dosing, exaggerated flinching responses and vocalizations were also noted in rats dosed at 10 mg/kg, which were considered treatment-related and consistent with acute local venous irritation.

In this study, the acute tolerated dose of SOC-1 administered as an intravenous bolus was determined to be at least 30 mg/kg.

Example 10 - 24-hour pharmacokinetic study of SOC-1 test item administered by intravenous bolus in Sprague Dawley rats

Three adult male Sprague Dawley rats were given a single intravenous bolus dose of 20 mg/kg (equivalent to the free base expressed as corrected purity) of the SOC-1 test item prepared in Hartman's solution. Ten blood samples were collected from each rat at time points from pre-dose to 24 hours post-dose. The blood samples were then processed to plasma, and the concentration of SOC-1 in plasma was determined using an LC-MS/MS method (assay range = 50-5000 ng/mL). Pharmacokinetic parameters were estimated from the plasma concentration-time data using a non-compartmental approach using Phoenix 64 software.

This study evaluated the 24-hour pharmacokinetics of SOC-1 in male Sprague Dawley rats after intravenous (i.v.) administration. The compound formulated in Hartman's solution (saline) was administered intravenously to a group of n=3 rats at a rate of 20 mg/kg (17.1 mg/kg, expressed as the free base corrected for purity). Ten blood samples were collected from each rat at time points from pre-dose to 24 hours post-dose. The blood samples were then processed into plasma and the concentration of SOC-1 was determined using an LC-MS/MS screening bioanalytical method.

The objective of this study was to investigate the pharmacokinetics of a single bolus dose of a SOC-1 test item administered by the intravenous route in adult Sprague Dawley rats.

This study was conducted in accordance with the guidelines set by the Australian Health and Medical Research Council, Australian Code of Practice for the Use of Animals for Scientific Purposes, 8th Edition, 2013 (1). This study was assessed and approved by the University of Queensland Animal Ethics Committee.

Test and vehicle/control items

The "Test Items", "Bioassay Reference Items" and "Vehicle/Control Items" are shown in Table 14, Table 15 and Table 16, respectively.

Table 14 Test items used in this embodiment

Table 15 Bioanalytical References Used in This Example

Table 16 Carriers/controls used in this example

Test and control formulations

The SOC-1 test item was dissolved in Hartmann's solution at a final concentration of 5 mg/mL for dosing. The formulations were prepared under clean conditions and in a laminar flow cabinet using sterile polypropylene tubes with gentle vortexing/swirl to ensure complete dissolution of the compound in the vehicle prior to use.

The test formulations were stored at ambient temperature and used on the same day. No dose formulation analytical samples were collected for this study.

animal

Table 17 shows details related to the animals used in the study.

Table 17 - Animals used in this example

The Sprague Dawley rat is considered an acceptable and commonly used species for pharmacokinetic studies.

The test system environment is shown in Table 18.

Table 18 - Test system environment used in this example

method

The pharmacokinetic study was conducted according to the UOS-004 study plan and the relevant test facility standard operating procedures (SOPs) as described below. The study design is shown in Table 19.

Table 19 - Study Design Used in This Example

^a) (Expressed as free base corrected for purity)

Prior to treatment, rats underwent surgery to cannulate the jugular vein (for dose administration, intravenous) and femoral artery (for blood sample collection, IA), with external cannula placement per the testing facility SOP. After recovery from surgery, rats were transferred to the Culex system for dose administration and blood sampling (care per the testing facility SOP).

The drug was administered intravenously via jugular vein cannula at a dosing volume of 4.0 mL/kg on the second day after surgery. Treatment was followed by vehicle administration to ensure that the full dose was delivered to the animal.

Blood samples were collected in lithium heparin tubes in IA cannulae at the following time points: pre-dose, 2, 5, 10, 30, 60 min, 2, 4, 8, and 24 h post-dose.

Immediately after collection, blood samples were stored on ice to minimize degradation and centrifuged as quickly as possible at approximately 4000 rcf for 10 minutes. Prior to dosing, pre-dose samples were centrifuged separately. Plasma was then transferred to polypropylene tubes and stored frozen at -80°C.

After the final sample was collected, the animals were euthanized by administering 325 mg/kg Lethabarb ^™ via IV cannula.

Study samples were subsequently transferred to dry ice for measurement of SOC-1 concentrations.

Body weight was collected immediately before surgery. Morbidity and mortality checks were completed daily during the acclimation period and at least twice during the study period.

SOC-1 plasma concentration analysis

Plasma concentrations of SOC-1 were determined using a screening LC-MS/MS method with the following liquid/liquid extraction. Briefly, after the addition of 10 μl of internal standard, a 50 μl aliquot of test sample and calibration standard was mixed with 1 mL of hexane:dichloromethane (1:1) for 5 minutes, then centrifuged and the organic phase decanted and dried under nitrogen. The sample was then reconstituted with 0.5 mL of 0.1% formic acid in 30% methanol in deionized water, and 0.2 mL was transferred to a 96-well plate. A 5 μl aliquot of this sample was then analyzed using an ABSciex API 4000 MS/MS coupled to a Shimadzu Prominence HPLC. The HPLC gradient consisted of 0.1% formic acid in deionized water (mobile phase A) and 0.1% formic acid in methanol (mobile phase B). Appendix 2 summarizes the LC-MS/MS instrument settings. Samples were analyzed with calibration samples (50-5000 ng/mL) and concentration data were calculated from the standard curve.

Plasma concentration data were exported using Analyst™ v1.6.1 (AB Sciex) software and then tabulated using GraphPad Prism 6 and/or Excel 2010.

The group mean ± standard error values of plasma concentration-time values were determined using Phoenix 64 WinNonlin software. The following pharmacokinetic parameters were also calculated by non-compartmental methods using Phoenix 64 WinNonlin software:

• Rsq – correlation coefficient of the terminal elimination phase (R ² ).

Lambda - the terminal elimination rate constant (1/h) associated with the terminal (log-linear) portion of the curve.

T _1/2 – terminal elimination half-life (h)

• C ₀ - Initial concentration at 0 minutes (ng/mL), determined by back extrapolation from the first two measured concentration values.

• _Tmax - the time of maximum plasma concentration after extravascular administration.

_Cmax - maximum plasma concentration after extravascular administration

AUC _last - Area under the curve from the time of dosing (dose time, 0 minutes) to the last measurable concentration (ng·h/mL).

AUC _inf - AUC (ng.h/mL) extrapolated to infinity based on the last measured concentration (obs).

AUC% Extra - The percentage of AUCinf extrapolated from the last measurable concentration to infinity.

Vz - distribution volume (L/kg).

Systemic clearance of Cl- (L/h/kg).

Note: If ^R2 < 0.85 and/or AUC% extrapolation > 20%, terminal elimination phase parameters are not reported.

Body weight and clinical symptoms

SOC-1 treatment was tolerated and no morbidity or mortality was observed. Table 20 shows the individual body weight values before treatment.

Table 20 - Body weight of rats used in this example

^a) Body weight was determined at the time of surgery one day before treatment.

^b) 17.1 mg/kg, expressed as the free base corrected for purity.

Plasma concentration vs. time data

Individual and mean plasma SOC-1 concentration-time results are shown in Table 21 and Figure 15.

Table 21 - SOC-1 plasma concentration-time relationship in rats in this example

^a) BLOQ = lower limit of quantitation (BLOQ) of less than 50 ng/mL.

SOC-1 plasma concentration-time data were determined following a single intravenous dose of 20 mg/kg (17.1 mg/kg expressed as the free base corrected for purity) to male Sprague Dawley rats.

Pharmacokinetic parameters

The pharmacokinetic parameters are summarized in Table 22.

Table 22 - SOC-1 pharmacokinetic parameters determined in this example

SOC-1 plasma concentration-time data were determined following a single intravenous bolus dose of 20 mg/kg (17.1 mg/kg expressed as free base corrected for purity) to male Sprague Dawley rats.

Briefly, following intravenous administration of 20 mg/kg SOC-1 (17.1 mg/kg, expressed as the purity-corrected free base), mean (± standard error of the mean (SEM)) values for _C0 and _AUCinf were 18200 (±2860) ng/mL and 17000 (±840) ng.h/mL, respectively. The mean (±SEM) value for _T1/2 was estimated to be 0.550 (±0.049) h, and the mean (±SEM) values for Vz and Cl were 0.799 (±0.051) L/kg and 1.01 (±0.051) L/h/kg, respectively.

The data show a monoexponential decrease in concentration over time. However, the observed decrease in plasma concentration may reflect only redistribution of SOC-1 rather than elimination, as the terminal elimination phase of SOC-1 may not have been captured. A more precise determination of the terminal elimination phase could be achieved using a method with improved sensitivity that enables quantification of sample concentrations over 4 hours.

Summarize

In this study, the pharmacokinetics of SOC-1 were characterized in adult male Sprague Dawley rats following a single intravenous dose of 20 mg/kg (expressed as the free base corrected for purity).

Example 11 - Acute Toxicity Evaluation of a Single Bolus Dose of 50 mg/kg of SOC-1 Test Item Administered Intravenously to Sprague-Dawley Rats in a One-Week Dose Ranging Study

The acute toxicity of the SOC-1 test item was evaluated in Sprague Dawley rats following intravenous administration of a single bolus dose of 50 mg/kg. A total of n=3 adult female rats were treated and then observed for 7 days before termination on day 8 of the study without necropsy.

In this study, administration of the SOC-1 test item at the 50 mg/kg dose level was well tolerated. Mild piloerection and decreased activity levels were noted on study day 1, which resolved within 4 hours and were considered treatment-related.

In this study, the acute tolerated dose of SOC-1 administered as an intravenous bolus was determined to be at least 50 mg/kg.

The nonclinical toxicity studies described herein entailed administering a single intravenous bolus of SOC-1 to adult Sprague Dawley rats, followed by one week of observation. Previous studies conducted in this testing facility have shown that rats can tolerate doses up to 30 mg/kg (1). The dose-ranging study described here involved administration of the test article at a dose of 50 mg/kg, which is considered the maximum feasible dose for the currently available 5 mg/mL formulation administered via the intravenous route.

The objectives of this study were to investigate the tolerability and acute toxicity of a single 50 mg/kg dose of the SOC-1 test item administered intravenously to adult Sprague Dawley rats.

This study was conducted in accordance with the guidelines set by the Australian Health and Medical Research Council and the Australian Code of Practice for the Care and Use of Animals for Scientific Purposes, 8th edition, 2013 (1). This study was assessed and approved by the University of Queensland Animal Ethics Committee.

Test and vehicle/control items

The "test items" and "vehicle/control items" are shown in Table 23 and Table 24, respectively.

Table 23 - Test items used in this example

Table 24 - Vectors/Controls used in this example

Test and control formulations

The SOC-1 test article was dissolved in Hartmann's solution at a final concentration of 5 mg/mL for dosing. The formulations were prepared under clean conditions and in a laminar flow cabinet using sterile polypropylene tubes with gentle vortexing/swirl to ensure complete dissolution of the compound in the vehicle prior to use.

The test formulations were stored at ambient temperature and used on the same day. No dose formulation analytical samples were collected for this study.

animal

Details regarding the animals used in the study are shown in Table 25.

Table 25 Animals used in this example

Sprague Dawley rats have been found to be an acceptable and commonly used species for toxicology studies; female rats were used in this study simply to minimize animal use.

The test system environment is shown in Table 26.

Table 26 - Test system environment used in this example

method

The design of the acute single-dose toxicity study in rats was adapted from the OECD Guideline for the Testing of Chemicals No. 420, Acute Oral Toxicity – Fixed-Dose Method (2001) (2) and the test facility standard operating procedure SP_T002 “Dose-Ranging Studies in Rodents”. The study procedures are described below and were conducted in accordance with the relevant test facility standard operating procedures.

The study design is summarized in Table 27.

Table 27 - Study Design Used in This Example

Briefly, after the acclimation period, the study began with dosing on study day 1. Animals were observed for signs of toxicity throughout the study as described below. Body weights were also collected daily, and on study day 8, surviving animals were sacrificed without necropsy.

treat

On day 1 of the study, conscious animals were restrained and treated with 5 mg/mL of the test item as an intravenous bolus at a dose volume of 10 mL/kg.

observe

Morbidity and mortality were recorded daily during the acclimation period and on day 8, and twice daily from day 1 to day 7.

Conduct at least one clinical observation during the acclimation period. Clinical observations should be conducted at least once from Day 1 until the day before termination. On the day of treatment, animals should be monitored continuously for the first 30 minutes after treatment, with formal observations conducted 30 minutes, 1 hour, and 4 hours after treatment.

Clinical observations include examining the animal's skin and coat, changes in the eyes and mucous membranes, respiratory and circulatory function, gait and posture, behavior, tremors or convulsions, and any other abnormalities.

Clinical observations also included daily inspection of the tail for any injection site reactions (eg, erythema or edema).

weight

Body weights were recorded daily during the acclimation period and from day 1 to day 8 of treatment.

autopsy

On day 8, all animals underwent final body weight measurement and were then euthanized by an overdose of pentobarbital (intraperitoneally). Carcasses were discarded without autopsy.

Data collection and evaluation

Data were collected and tabulated manually using GraphPad Prism 6 and/or Excel 2010.

Morbidity and mortality

There were no unplanned deaths or morbidity events in this study.

Clinical observation

Rats treated with 50 mg/kg SOC-1 test item were found to have mild piloerection and decreased activity levels on study day 1. These findings were generally detected within 30 minutes of treatment and resolved within 4 hours after dosing. These findings were considered to be related to treatment. One rat was also found to have a hunched posture or gait on study day 5, which was considered incidental and unrelated to treatment. No injection site reactions were observed, and clinical observations are shown in Table 28.

Table 28 - Summary of the incidence of clinical signs in rats in this example

Body weight increased or remained stable in all animals during the study period. No weight changes were considered treatment-related.

The body weight values are summarized in Figure 16 and Table 29, and the individual body weight values are shown in Table 30.

Table 29 - Average body weight of rats in this example ± standard deviation (SD)

Study Days Weight (g) 1 224.30±7.45 2 223.07±4.37 3 226.87±4.33 4 223.03±5.34 5 222.77±4.75 6 223.80±3.70 7 226.60±2.57 8 228.03±3.81

Table 30 - Body weight values of individual rats used in this example

Weight values are in grams.

in conclusion

In this study, treatment with the SOC-1 test item was tolerable at a dose level of 50 mg/kg. Mild piloerection and decreased activity levels were noted after dosing on study day 1, findings that resolved within 4 hours and were considered treatment-related.

In a previous study in Sprague Dawley rats, intravenous administration of 10 mg/kg SOC-1 prepared in saline was associated with abnormal vocalization and flinching responses at the time of dosing (1). Administration of 50 mg/kg SOC-1 prepared in Hartman's solution was not associated with similar responses in the current study.

In this study, the acute tolerated dose of SOC-1 administered as an intravenous bolus was determined to be at least 50 mg/kg.

Example 12 - Assessment of Alcohol Intake in Baboons Administered with SOC-1

animal

Baboons are considered ideal subjects for these studies because the technical field can provide sufficient data on the effects of alcohol in baboons, as well as their responses to feeding manipulations, and the manipulation conditions are well studied. The inventors chose to use baboons as a model of alcohol dependence because baboons can drink large amounts of alcohol and can be trained to work for alcohol and palatable food rewards. In addition, non-human primates have a higher homology with the human brain than rodents.

The baboons were obtained from World Wide Primates in Florida. The four baboons used in the study were part of a larger group used in other studies but had not participated in other types of research in over a year. The four included here were selected from an original group of eight. These four were willing to drink heavily and were chosen for the study protocol. Previous studies of these baboons included responses to food and drugs such as methamphetamine. Previous studies did not include any surgical procedures or induction of dependence on drugs of abuse.

The four baboons had a history of moderate alcohol consumption. The animals had participated in a previous program that provided them with limited exposure to alcohol. During this period, on intermittent days, they were willing to consume between 0.9 and 1.5 g/kg of alcohol over a two-hour period.

Baboons are housed in a novel enclosure consisting of a standard extruded back canopy with a large solid bench attached front to back, connected to a custom cage extension. The baboons sit either underneath or on top of the habitat. Sitting on top of the habitat is the preferred position for most animals. This arrangement allows the animals to walk between cages, use the habitat, use the bench, place their feet on the ledge on the top half of each cage, or sit on the floor. This innovative enclosure allows us to provide a stable housing environment in which to maintain healthy nonhuman primates for many years for long-term behavioral and imaging studies.

Alcohol self-administration

Baboons had been trained to self-administer 4-8% alcohol (w/v) using the sugar-addition alcohol method. Baboons were habituated to lever-pressing rewards and had previously been trained to respond to sweetened Kool-Aid. Once they responded to Kool-Aid (mixed to 1/4 the package insert strength), ethanol (4-8% w/v) was added to the Kool-Aid. Fluid delivery was accompanied by a tone and the illumination of a stimulus light above the fluid spout.

On each day of the study, voluntary alcohol consumption was administered using a fixed-ratio schedule with alcohol available over a 2-hour period. The first session began at 9:00 AM, and the second session began at 1:00 PM. Baboons received their daily rations at noon each day. Each session consisted of a 26-minute trial, during which a light above the alcohol lever indicated the availability of alcohol. Baboons received an aliquot of alcohol (4% w/v, 5 ml) by pulling the lever 10 times (FR10), with 5 aliquots available per trial. The trial ended when the baboon received 5 reinforcers or had no response for 6 minutes. There was a 1-minute inter-trial interval during which no alcohol was available. Thus, 100 drinks, or 500 ml, were available in the mornings and afternoons Monday through Friday.

SOC-1 administration

During the study, baboons were given oral doses of 1-8 mg/kg of SOC-1. The drug was delivered to the baboons as a powder, with the correct dose placed in the center of a fig along with some sugar. These figs were then fed to the baboons 60 minutes before the daily dosing phase. The drug or placebo was administered acutely no more than twice a week.

The results of the study are shown in Figures 17, 18, and 19. Figure 17 shows the total amount of fluid delivered as a function of SOC-1 dose. Figures 18 and 19 show the results for morning and afternoon alcohol intake, respectively.

Example 13 - Assessment of Autism Spectrum Disorders in the BALB/C Mouse Model

Compared to standard C57BL/6 mice, BALB/C mice have autism-like behaviors (including reduced interest in social stimuli). In a social preference test, BALB/C mice showed no preference for interacting with object stimuli (novel objects) versus social stimuli (novel mice), while C57BL/6 mice showed a strong preference. SOC-1 (5 mg/kg IP) significantly increased the time BALB/C mice spent investigating social stimuli until they were no longer distinguishable from standard strain C57BL/6 mice. SOC-1 (5 mg/kg IP) also significantly increased the time BALB/C mice spent investigating object stimuli, suggesting that in addition to increasing interest in social stimuli, it can also cause a more general increase in exploratory behavior. The results of the study are shown in Figure 20.

It will be appreciated by those skilled in the art that various changes and/or modifications may be made to the above embodiments without departing from the broad general scope of the present disclosure. Therefore, the present embodiments are to be considered in all aspects as illustrative and not restrictive.

Claims (12)

1. A pharmaceutical composition comprising: a pharmaceutically acceptable carrier, diluent, or excipient; and a compound with the following structural structure, or a pharmaceutically acceptable salt thereof: 2. A pharmaceutical composition comprising: a pharmaceutically acceptable carrier, a diluent or excipient; and a compound with the following structure: 3. A pharmaceutical composition comprising: a pharmaceutically acceptable carrier, a diluent or excipient; and a pharmaceutically acceptable salt of a compound with the following structural composition: 4. The pharmaceutical composition of claim 3, wherein the pharmaceutically acceptable salt of the compound is selected from any one of the following: hydrochloric acid, sulfuric acid, phosphoric acid, nitric acid, carbonic acid, boric acid, aminosulfonic acid, hydrobromic acid, acetic acid, propionic acid, butyric acid, tartaric acid, maleic acid, hydroxymaleic acid, fumaric acid, hydroxyethanesulfonic acid, malic acid, citric acid, lactic acid, mucoic acid, gluconic acid, benzoic acid, succinic acid, oxalic acid, phenylacetic acid, methanesulfonic acid, toluenesulfonic acid, benzenesulfonic acid, salicylic acid, sulfanilic acid, stearic acid, palmitic acid, oleic acid, lauric acid, pantothenic acid, tannic acid, ascorbic acid, or valerate; or a metal salt; or an ammonium or alkylammonium salt; or an amino acid salt. 5. The pharmaceutical composition according to any one of claims 1-4, characterized in that the pharmaceutical composition is formulated as a composition selected from the group consisting of: oral administration composition, rectal administration composition, nasal administration composition, topical administration composition, parenteral administration composition, inhalation administration composition, or spray administration composition. 6. Use of a compound with the following structure or a pharmaceutically acceptable salt thereof; Used to prepare a medicament for treating or preventing social dysfunction in a subject; treating or preventing mental disorders in a subject; and/or treating a subject who has suffered from or is recovering from substance abuse disorder and is attempting to maintain sustained abstinence from the medicament. 7. Uses of a compound with the following structure; Used to prepare a medicament for treating or preventing social dysfunction in a subject; treating or preventing mental disorders in a subject; and/or treating a subject who has suffered from or is recovering from substance abuse disorder and is attempting to maintain sustained abstinence from the medicament. 8. Use of a pharmaceutically acceptable salt of a compound with the following structure; Used to prepare a medicament for treating or preventing social dysfunction in a subject; treating or preventing mental disorders in a subject; and/or treating a subject who has suffered from or is recovering from substance abuse disorder and is attempting to maintain sustained abstinence from the medicament. 9. The use as described in claim 8, characterized in that the pharmaceutically acceptable salt of the compound is selected from any one of the following groups: hydrochloric acid, sulfuric acid, phosphoric acid, nitric acid, carbonic acid, boric acid, aminosulfonic acid, hydrobromic acid, acetic acid, propionic acid, butyric acid, tartaric acid, maleic acid, hydroxymaleic acid, fumaric acid, hydroxyethanesulfonic acid, malic acid, citric acid, lactic acid, mucoic acid, gluconic acid, benzoic acid, succinic acid, oxalic acid, phenylacetic acid, methanesulfonic acid, toluenesulfonic acid, benzenesulfonic acid, salicylic acid, sulfanilic acid, stearic acid, palmitic acid, oleic acid, lauric acid, pantothenic acid, tannic acid, ascorbic acid, or valerate; or metal salt; or ammonium, alkylammonium salt; or amino acid salt. 10. The use according to any one of claims 6-9, characterized in that the pharmaceutical composition is formulated as a composition selected from the group consisting of: oral administration composition, rectal administration composition, nasal administration composition, topical administration composition, parenteral administration composition, inhalation administration composition, or spray administration composition. 11. The pharmaceutical composition of claim 3, wherein the pharmaceutically acceptable salt of the compound is a salt of aspartic acid. 12. The use as described in claim 8, wherein the pharmaceutically acceptable salt of the compound is a salt of aspartic acid.