The present application claims priority from Australian Provisional Patent Application No. 2020903196 filed on 7 Sep. 2020, the entire contents of which is incorporated herein by reference.
This invention relates to salts and crystals of 1-methyl-1,4,5,10-tetrahydropyrazolo[3,4-b][1,5]benzodiazepine.
1-Methyl-1,4,5,10-tetrahydropyrazolo[3,4-b][1,5]benzodiazepine dihydrochloride is described in WO2017/004674 as possessing biological activity similar to oxytocin agonists, without demonstrating significant binding affinity for the orthosteric oxytocin receptor binding sites or the orthosteric vasopressin receptor binding sites. As such, there is interest in developing pharmaceuticals comprising this compound.
1-Methyl-1,4,5,10-tetrahydropyrazolo[3,4-b][1,5]benzodiazepine has the following structure:
This compound may also be referred to as 1-methyl-1,4,5,10-tetrahydrobenzo[b]pyrazolo[3,4-e][1,4]diazepine. References to 1-methyl-1,4,5,10-tetrahydropyrazolo[3,4-b][1,5]benzodiazepine and 1-methyl-1,4,5,10-tetrahydrobenzo[b]pyrazolo[3,4-e][1,4]diazepine are intended to be interchangeable as used herein.
While able to induce promising biological activity, in further development of 1-methyl-1,4,5,10-tetrahydropyrazolo[3,4-b][1,5]benzodiazepine dihydrochloride it was discovered that this salt form demonstrated high hygroscopicity. Dynamic vapour sorption (DVS) analysis of the material indicated a form change above 60% relative humidity (RH) with the weight change reversible below 30% RH. While remaining a useful laboratory investigative tool, the high hygroscopicity makes further development of the dihydrochloride salt unsuitable.
There is therefore a need to provide alternative forms of 1-methyl-1,4,5,10-tetrahydropyrazolo[3,4-b][1,5]benzodiazepine for incorporation into pharmaceutical products. Advantageously, the alternative forms would provide material with lower hygroscopicity at 60% RH and above than the 1-methyl-1,4,5,10-tetrahydropyrazolo[3,4-b][1,5]benzodiazepine dihydrochloride salt.
All publications, patents and patent applications that may be cited herein are hereby incorporated by reference in their entirety.
Reference to any prior art in the specification is not an acknowledgment or suggestion that this prior art forms part of the common general knowledge in any jurisdiction or that this prior art could reasonably be expected to be understood, regarded as relevant, and/or combined with other pieces of prior art by a skilled person in the art.
The invention provides solid forms of 1-methyl-1,4,5,10-tetrahydropyrazolo[3,4-b][1,5]benzodiazepine (“compound of the invention”) possessing lower hygroscopicity than previously described pharmaceutically acceptable forms of the compound of the invention, including the dihydrochloride salt of the compound of the invention. The solid forms described herein may also demonstrate improved thermal stability compared to previously explored forms of the compounds and provide at least substantially equivalent biologically available compound of the invention to a subject following administration.
Surprisingly, the inventors have found that phosphoric acid and L-tartaric acid addition salts of the compound of the invention, as well as a crystalline form of the freebase of the compound of the invention possess one or more of these improved properties.
In one aspect, the invention provides a phosphoric acid addition salt of the compound of the invention. This phosphoric acid addition salt may alternatively be referred to as a phosphate salt of the compound of the invention.
In another aspect, the invention provides an L-tartaric acid addition salt of the compound of the invention. This L-tartaric acid addition salt may alternatively be referred to as an L-tartrate salt of the compound of the invention.
In some embodiments, the phosphate and/or L-tartrate salts of the compound of the invention are in a crystalline form.
In a further aspect, the invention provides a crystalline form of the compound of the invention. This crystalline form may also be referred to as a crystalline form of a freebase of the compound of the invention.
In another aspect, the invention provides a solid form, typically a crystalline form, of the compound of the invention selected from:
Also described herein are methods of using these salt and crystalline forms of the compound of the invention, including in methods of:
As used herein, except where the context requires otherwise, the term “comprise” and variations of the term, such as “comprising”, “comprises” and “comprised”, are not intended to exclude further additives, components, integers or steps.
It must be noted that as used herein and in the appended claims, the singular forms “a”, “an” and “the” include plural reference unless the context clearly dictates otherwise. Thus, for example, a reference to “a symptom” and/or “at least one symptom” may include one or more symptoms, and so forth.
The term “and/or” can mean “and” or “or”.
The term “(s)” following a noun contemplates the singular or plural form, or both.
Various features of the invention are described with reference to a certain value, or range of values. These values are intended to relate to the results of the various appropriate measurement techniques, and therefore should be interpreted as including a margin of error inherent in any particular measurement technique. Some of the values referred to herein are denoted by the term “about” to at least in part account for this variability. The term “about”, when used to describe a value, may mean an amount within ±10%, 5%, ±1% or ±0.1% of that value.
Further aspects of the present invention and further embodiments of the aspects described in the preceding paragraphs will become apparent from the following description, given by way of example and with reference to the accompanying drawings.
The invention relates to salt and/or crystalline forms of the compound of the invention. These salt and/or crystalline forms include:
Collectively, the above salt and crystalline forms of the compound of the invention are referred to herein as the salts and/or crystals of the invention.
Each of the salts and/or crystals of the invention were surprisingly found to have desirable properties in terms of their low hygroscopicity while retaining bioavailability of the compound of the invention. The salt and/or crystalline forms described herein were the only forms of the compound of the invention possessing these properties from a screen of 18 acid counterions and 5 solvent systems (see Example 1).
The salts and/or crystals of the invention may be substantially non-hygroscopic when exposed to an environment with a minimum relative humidity of at least about 60% RH, 70% RH, 75% RH or 80% RH. The salts and/or crystals of the invention may be substantially non-hygroscopic when exposed to environments at a maximum relative humidity of not more than about 90%, 85%, 80% or 75%. The salts and/or crystals of the invention may be substantially non-hygroscopic when exposed to an environment having a relative humidity from any of these minimum values to any of these maximum values provided the minimum value is less than the maximum value. For example, in some embodiments, the salts and/or crystals of the invention are substantially non-hygroscopic when exposed to an environment at a relative humidity from about 60% to about 90% or from about 75% to about 85%. At relative humidities of about 90% RH and above the salts and/or crystals of the invention may increase in mass by no more than about 2 wt %, 1.5 wt %, 1 wt %, 0.9 wt %, 0.8 wt %, 0.7 wt %, 0.6 wt % or 0.5 wt % due to the absorption of water. The increase in mass may be measured by DVS, for example, according to any procedure described herein.
The salts and/or crystals of the invention may also be substantially stable for an extended period of time. For example, the salts and/or crystals may be stable for a period of 1 week, 1, 2, 3, 4, 5, 6 months or longer upon storage at 25° C. and 60% RH. The salts and/or crystals may also be stable for 1 week, 1, 2, 3, 4, 5, 6 months or longer upon storage under accelerated storage conditions, for example, at 40° C. at 75% RH. In some embodiments, the salts and/or crystals retain at least about 95%, 96%, 97%, 98%, 98.5% or 99% purity upon storage under any of these storage conditions.
The salts and/or crystals of the invention may be prepared from 1-methyl-1,4,5,10-tetrahydropyrazolo[3,4-b][1,5]benzodiazepine prepared by any suitable means. The synthesis of 1-methyl-1,4,5,10-tetrahydropyrazolo[3,4-b][1, 5]benzodiazepine and its dihydrochloride salt have been previously described, including in WO2017/004674 (U.S. Ser. No. 11/033,555) which is incorporated herein entirely by reference.
In some embodiments, the salts and/or crystals of the invention are prepared from 1-methyl-1,4,5,10-tetrahydropyrazolo[3,4-b][1,5]benzodiazepine prepared by reacting N-(1-methyl-1H-pyrazol-5-yl)-benzene-1,2-diamine with formaldehyde in a solvent and in the presence of an acid to provide 1-methyl-1,4,5,10-tetrahydropyrazolo[3,4-b][1,5]benzodiazepine. Any acid capable of mediating the reaction may be used. Suitable acids include acetic acid, phosphoric acid, and so on. As will be discussed further below, when phosphoric acid is included in this reaction step, the product may be the phosphoric acid addition salt of the compound of the invention.
In some embodiments, the salts and/or crystals of the invention are prepared from 1-methyl-1,4,5,10-tetrahydropyrazolo[3,4-b][1,5]benzodiazepine prepared by the following steps:
In a first aspect, the invention provides a phosphoric acid addition salt of 1-methyl-1,4,5,10-tetrahydropyrazolo[3,4-b][1,5]benzodiazepine. This salt form may be referred to herein as the phosphate salt of the invention.
The phosphate salt of the invention may be a hydrogen phosphate, dihydrogen phosphate or a phosphate salt of the compound of the invention. In some embodiments, the phosphate salt is a dihydrogen phosphate salt of the compound of the invention.
Typically, the phosphate salt of the invention is crystalline. It has been found that crystalline forms of the phosphate salt of the invention demonstrate polymorphism, with 2 distinct polymorphs identified, referred to herein as phosphate form 1 and phosphate pattern 2. It has also been found that phosphate form 1 is a stable crystalline form of the phosphate salt of the invention, while form 2 is unstable and converts to form 1 over time. Therefore, in some embodiments, the phosphate salt of the invention is provided as phosphate form 1.
Phosphate form 1 may be characterised by its X-ray diffraction (XRD) pattern. The XRD pattern for phosphate form 1 comprises characterising strong peaks at about 12° and about 18° 2θ.
Additionally, phosphate form 1 may be characterised by peaks in the XRD pattern at about 12°, 14°, 17.5°, 18°, 19°, 20°, 20.8°, 22.5°, 24°, 24.8°, 26°, 26.5° and 27.8° 2θ.
Typically, phosphate form 1 may be characterised by the XRD pattern shown in
Phosphate form 1 may additionally or alternatively be characterised by its melting point. It has been found that the melting point of phosphate form 1 (about 200° C.) is higher by about 8° C. than the melting point for phosphate pattern 2 indicating that it has a higher degree of crystallinity and hence greater stability. DT analysis of phosphate pattern 2 showed a broad melting endotherm from an onset of about 201° C., with a peak at 206° C., which was followed immediately by thermal degradation. In contrast, DT analysis showed a large endothermic melting transition for phosphate form 1 from an onset of about 209° C. with a peak at 214° C. Therefore, in some embodiments the phosphate salt of the compound of the invention may have a melting point of about 200° C.
The phosphate salt of the compound of the invention is typically an anhydrous crystal. The anhydrous nature of this crystal form may be determined by thermogravimetric analysis.
The phosphate salt of the invention may remain substantially anhydrous and at purity levels that are substantially unchanged when stored under ambient conditions or under accelerated storage conditions. The accelerated storage conditions may comprise elevated temperature (eg 40° C. or 80° C.) and/or increased relative humidity. In some embodiments, the phosphate salt may remain substantially anhydrous upon storage for example for at least 1, 2 or 3 week(s), 1, 2, 3, 4, 5, 6 months or longer, at elevated temperature (eg 40° C.) and at up to about 60% RH, 70% RH or 75% RH. Typically, the phosphate salt will also remain stable under these storage conditions, remaining substantially pure throughout, for example resulting in up to about 2%, 1.5% or 1% degradation products detectable by HPLC. The HPLC may be carried out by any of the techniques described herein.
The phosphate salt of the invention may be prepared by any suitable means. The process may involve combination of phosphoric acid with the compound of the invention in a suitable solvent. This process may be carried out on isolated freebase material, or may be conducted in a 1-pot process with the final synthetic step of preparing the compound of the invention, where the salt is formed without isolating the freebase.
Extensive solvent screening was carried out to determine what conditions influenced formation of crystalline forms—form 1 and pattern 2 (see example 4). It was found that form 1 is the form that is provided under most conditions except in highly polar solvents such as water, N-methyl pyrrolidine (NMP) and dimethylsulfoxide (DMSO) due to the salt's solubility, when left to stand in concentrated solutions of ethyl acetate, methylisobutyl ketone and tert-butylmethylether having been subjected to heating cooling cycles, or when allowed to mature in tert-butylmethylether. Mixtures of form 1 and pattern 2 were obtained when ethyl formate (maturation and standing following heat cycling), isopropyl acetate (standing following heat cycling), methylethyl ketone (standing following heat cycling), and chloroform (trace pattern 2 on standing following heat cycling).
Accordingly, also provided is a process for preparing the phosphate salt of the invention, comprising
In some embodiment, the method further comprises preparing a crystallisation solution of the phosphate salt and a minimum volume of a crystallisation solvent to form the crystallisation solution. The crystallisation solution may be allowed to stand under ambient conditions and/or cooled and/or concentrated to allow crystal formation.
In embodiments where form 1 is desired, the crystallisation solvent typically does not comprise ethyl acetate, methylisobutyl ketone, tert-butylmethyl ether, ethyl formate, isopropyl acetate, and methyl ethyl ketone or a combination thereof. In some embodiments, the solvent further does not comprise chloroform.
When form 1 is desired, the crystallisation solvent may be selected from 1,4-dioxane, 2-butanol, 2-ethoxyethanol, 2-methyl tetrahydrofuran, 2-propanol, acetone, acetonitrile, methanol, anisole, ethanol, tetrahydrofuran, ethyleneglycol and water or a combination thereof.
When phosphate pattern 2 is desired, the crystallisation solvent is preferably tert-butylmethylether.
Also provided is a process for preparing the phosphate salt of the invention, comprising reacting N-(1-methyl-1H-pyrazol-5-yl)-benzene-1,2-diamine with formaldehyde in the presence of phosphoric acid in a solvent to provide the phosphoric acid addition salt of the compound of the invention.
The reaction of N-(1-methyl-1H-pyrazol-5-yl)-benzene-1,2-diamine with formaldehyde may be carried out in any suitable solvent, such as any of the crystallisation solvents described herein. The reaction of N-(1-methyl-1H-pyrazol-5-yl)-benzene-1,2-diamine with formaldehyde may further comprise forming a solution of N-(1-methyl-1H-pyrazol-5-yl)-benzene-1,2-diamine and phosphoric acid in a solvent, and adding formaldehyde to the solution. The solution may comprise any suitable solvent. In some embodiments, the solvent is an aqueous solvent. In some embodiments, the solvent is selected from 1,4-dioxane, 2-butanol, 2-ethoxyethanol, 2-methyl tetrahydrofuran, 2-propanol, acetone, acetonitrile, methanol, anisole, ethanol, tetrahydrofuran, ethyleneglycol and water or a combination thereof. In some embodiments, the solvent is selected from acetonitrile, water or a combination thereof. Combinations of solvents may comprise any suitable mixture of components, for example a 2 solvent mixture such as acetonitrile and water may be in a ratio by weight of from about 1:1 to about 2:1 acetonitrile to water.
The reaction of N-(1-methyl-1H-pyrazol-5-yl)-benzene-1,2-diamine with formaldehyde may progress at elevated temperatures. In some embodiments, the temperature of the reaction is from about 25° C. to about 50° C., about 25° C. to about 45° C. or about 35° C. to about 45° C. In some embodiments, the temperature of this reaction may be carried out at a temperature of about 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44 or 45° C.
The reaction temperature may be from any of these temperatures to any other of these temperatures.
The reaction of N-(1-methyl-1H-pyrazol-5-yl)-benzene-1,2-diamine with formaldehyde may include any suitable amount of phosphoric acid. Typically, the phosphoric acid is present in this step in an amount of about 1 molar equivalent with respect to the N-(1-methyl-1H-pyrazol-5-yl)-benzene-1,2-diamine (and hence the reaction product). In some embodiments, the phosphoric acid is provided in a molar excess relative to the N-(1-methyl-1H-pyrazol-5-yl)-benzene-1,2-diamine, such as at least about 1, 1.05, 1.1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5 equivalents or more. The molar equivalents of the phosphoric acid may be from any of these values to any other of these values, for example, from about 1 to about 5 equivalents phosphoric acid relative to N-(1-methyl-1H-pyrazol-5-yl)-benzene-1,2-diamine.
The phosphate salt of the invention produced in these methods may be crystalline, eg phosphate form 1. However, in some embodiments, the process may further comprise a a step of forming a crystallising solution comprising the phosphoric acid addition salt, which may be carried out according to any such step described herein.
Following the reacting step, the processes typically comprise separating excess solvent and phosphoric acid to provide the phosphate salt. In some embodiments, the separation may be achieved by filtration.
In some embodiments, the process for preparing the phosphate salt of the invention may comprise:
In a second aspect, the invention provides an L-tartaric acid addition salt of 1-methyl-1,4,5,10-tetrahydropyrazolo[3,4-b][1,5]benzodiazepine. This salt form may be referred to herein as the L-tartrate salt of the invention.
Typically, the L-tartrate salt of the invention is provided in a crystalline form. The crystalline form may be characterised by XRD. Accordingly, in some embodiments the L-tartrate salt is characterised by the XRD shown in
The L-tartrate salt of the invention may additionally or alternatively be characterised by its melting point. In some embodiments, the melting point of the L-tartrate salt of the invention is about 181° C.
The L-tartrate salt of the invention may be an anhydrous crystal. The anhydrous nature of this crystal form may be determined by TGA.
The L-tartrate salt of the compound of the invention may remain substantially anhydrous and at purity levels that are substantially unchanged when stored under ambient conditions or under accelerated storage conditions. The accelerated storage conditions may comprise elevated temperature (eg 40° C. or 80° C.) and/or increased relative humidity. In some embodiments, the L-tartrate salt may remain substantially anhydrous upon storage for example for at least 1 week, at elevated temperature (eg 40° C.) and at up to about 60% RH, 70% RH or 75% RH. Typically, the L-tartrate salt will also remain stable under these storage conditions, remaining substantially pure throughout, for example resulting in up to about 2%, 1.5% or 1% degradation products detectable by HPLC. The HPLC may be carried out by any of the techniques described herein.
The L-tartrate salt of the invention may be prepared by any suitable means. Typically, the preparation of the L-tartrate salt of the invention comprises exposing the compound of the invention to L-tartaric acid.
Accordingly, also provided is a process for preparing the L-tartrate salt of the invention, comprising:
In some embodiments, the process further comprises preparing a crystallisation solution of the L-tartrate salt and a minimum volume of a crystallisation solvent to form the crystallisation solution. The crystallisation solution may be allowed to stand under ambient conditions and/or cooled and/or concentrated to allow crystal formation.
Also provided is a process for preparing the L-tartrate salt of the invention comprising reacting N-(1-methyl-1H-pyrazol-5-yl)-benzene-1,2-diamine with formaldehyde in the presence of L-tartaric acid to provide the L-tartaric acid addition salt of the compound of the invention.
The reaction of N-(1-methyl-1H-pyrazol-5-yl)-benzene-1,2-diamine with formaldehyde may occur in any suitable solvent, such as any of the crystallisation solvents described herein. The reaction of N-(1-methyl-1H-pyrazol-5-yl)-benzene-1,2-diamine with formaldehyde may further comprise forming a solution of N-(1-methyl-1H-pyrazol-5-yl)-benzene-1,2-diamine and adding formaldehyde to the solution. The solution may comprise any suitable solvent, such as any of the crystallisation solvents for forming tartrate salts described herein.
The phosphate salt of the invention produced in these methods may be crystalline. However, in some embodiments, the process may further comprise a step of forming a crystallising solution comprising the L-tartaric acid addition salt, which may be carried out according to any such step described herein.
In some embodiments, the process for preparing the L-tartaric salt of the invention may comprise:
In a third aspect, the invention provides a crystal of the compound of the invention. This crystal may be referred to herein as the freebase crystal of the invention.
Typically, the freebase crystal of the invention is an anhydrous crystal.
The freebase crystal of the invention may be characterised by its XRD pattern, which is shown in
The freebase crystal of the invention may be prepared by any suitable means. Typically, the preparation of the freebase crystal of the invention comprises exposing a salt of the compound of the invention (such as a hydrochloride salt of the compound of the invention) to an aqueous base (such as sodium bicarbonate) to neutralise the acid addition counterion, followed by liquid-liquid extraction with an organic solvent to extract the freebase compound in an organic phase.
Accordingly, also provided is a process for preparing a crystalline form of 1-methyl-1,4,5,10-tetrahydropyrazolo[3,4-b][1,5]benzodiazepine freebase, comprising:
Also provided is a process for preparing a crystalline form of 1-methyl-1,4,5,10-tetrahydropyrazolo[3,4-b][1,5]benzodiazepine freebase, comprising providing a solution of 1-methyl-1,4,5,10-tetrahydropyrazolo[3,4-b][1,5]benzodiazepine and allowing the 1-methyl-1,4,5,10-tetrahydropyrazolo[3,4-b][1,5]benzodiazepine to crystalise, wherein the solution is substantially free of acid.
In these methods, the 1-methyl-1,4,5,10-tetrahydropyrazolo[3,4-b][1,5]benzodiazepine may be provided by any suitable means, including by synthesis, including the synthesis described herein.
Also provided is a process for preparing a crystalline form of 1-methyl-1,4,5,10-tetrahydropyrazolo[3,4-b][1,5]benzodiazepine freebase, comprising reacting N-(1-methyl-1H-pyrazol-5-yl)-benzene-1,2-diamine with formaldehyde in a solvent in the presence of an acid, followed by exposing the reaction products to a base, and optional crystalising to provide the crystaline form of 1-methyl-1,4,5,10-tetrahydropyrazolo[3,4-b][1,5]benzodiazepine freebase.
In some embodiments, the acid is acetic acid.
In some embodiments, the base is an aqueous base, such as an aqueous solution of sodium bicarbonate, sodium carbonate, sodium hydroxide, potassium carbonate and the like. The exposing step may comprising multiple washes of the reaction products with the base.
The product following exposure to the base may be in crystaline form, or the process may require a subsequent crystalising step. The crystalising step may comprise providing a solution of 1-methyl-1,4,5,10-tetrahydropyrazolo[3,4-b][1,5]benzodiazepine and allowing the 1-methyl-1,4,5,10-tetrahydropyrazolo[3,4-b][1, 5]benzodiazepine to crystalise. Any suitable crystalising step described herein may be used in these processes.
Also provided are medicaments comprising any one or more of the salts and/or crystals of the invention.
Also provided are pharmaceutical compositions comprising any one or more of the salts and/or crystals of the invention. The pharmaceutical compositions typically further comprise a pharmaceutically acceptable carrier, diluent and/or excipient.
The medicaments and pharmaceutical compositions include those for oral, rectal, nasal, topical (including buccal and sub-lingual), parenteral administration (including intramuscular, intraperitoneal, sub-cutaneous and intravenous), or in a form suitable for administration by inhalation or insufflation. The salts and/or crystals of the invention optionally together with a conventional adjuvant, carrier or diluent, may thus be placed into the form of pharmaceutical compositions and unit dosages thereof, and in such form may be employed as solids, such as tablets or filled capsules, or liquids as solutions, suspensions, emulsions, elixirs or capsules filled with the same, all for oral use, or in the form of sterile injectable solutions for parenteral (including subcutaneous) use. Typically, the salts and/or crystals of the invention will be employed as solids due to their favourable properties in the solid state.
The pharmaceutical compositions of the salts and/or crystals of the invention may conveniently be presented in dosage unit form and may be prepared by any of the methods well known in the art of pharmacy and may include any conventional carrier, diluent and/or excipient as known in the art of pharmacy (See, for example, Remington: The Science and Practice of Pharmacy, 21st Ed., 2005, Lippincott Williams & Wilkins). Typically, preparation of the pharmaceutical compositions described herein include the step of bringing the active ingredient, for example any one of the salts and/or crystals of the invention, into association with the carrier which constitutes one or more accessory ingredients. In general, the pharmaceutical compositions are prepared by uniformly and intimately bringing the active ingredient, for example salts and/or crystals of the invention, into association with a liquid carrier or a finely divided solid carrier or both, and then, if necessary, shaping the product into the desired formulation. In the pharmaceutical composition the salts and/or crystals of the invention are included in an amount sufficient to produce the desired effect.
The pharmaceutical compositions described herein may be used in any of the methods described herein.
Methods of treatment involving the compound of the invention are described in WO2017/004674, WO2020/102857 and WO2021/042178. As the salts and/or crystals of the invention are favourable solid forms, possessing low hygroscopicity and improved stability, it is envisaged that they may be used in any of these methods of treatment.
Accordingly, in another aspect there is provided a method of:
In another aspect, also provided is a method of treating a subject suffering from or at risk of developing a substance abuse disorder, or a subject recovering from a substance abuse disorder and seeking to maintain ongoing abstinence of the substance, comprising administering to the subject an effective amount of any one or more of the salts and/or crystals of the invention, to thereby treat or prevent the substance abuse disorder.
In some embodiments, the method of the invention comprises administering an effective amount of a pharmaceutical composition comprising salts and/or crystals of the invention and a pharmaceutically acceptable carrier, diluent and/or excipient.
In some embodiments, the method of treating or preventing antisocial behaviour in a subject comprises stimulating pro-social behaviour in the subject.
In some embodiments, the psychiatric disorder is selected from autism spectrum disorder, substance abuse disorder, schizophrenia, or a combination thereof.
In some embodiments, the substance abuse disorder is selected from addiction and/or dependence on any one of an opioid, an opiate, alcohol, cocaine or a combination thereof.
In some embodiments, the methods of the invention treat a symptom of opioid withdrawal. The symptoms of opioid withdrawal include psychological, physical and/or somatic symptoms.
Physical and somatic symptoms of opioid withdrawal include tremors, shaking, hot or cold flashes, goosebumps, sweating, rapid breathing, elevated heart rate, elevated blood pressure, body aches, vomiting, diarrhea and fever. In some embodiments, methods treat a physical and/or somatic symptom of opioid withdrawal. In some embodiments, the physical and/or somatic symptoms are selected from tremors and shaking.
Psychological symptoms of opioid withdrawal include dysphoria, anxiety, restlessness, irritability, insomnia, yawning, hallucinations, hyperalgesia, hyperkatifiteia, and anorexia. It is believed that although these symptoms are not physical/somatic, they are symptoms of opioid withdrawal and stem from the physiological changes resulting from cessation or reduction of opioid dosing and/or induced by opioid antagonist administration. In some embodiments, the methods treat dysphoria.
Symptoms of opioid withdrawal include dysphoria, anxiety, restlessness, irritability, insomnia, yawning, hallucinations, tremors, shaking, hot or cold flashes, goosebumps, sneezing, sweating, rapid breathing, elevated heart rate, elevated blood pressure, pupillary dilation, piloerection, head aches, body aches, muscle cramps, muscle aches, bone aches, joint aches, hyperalgesia, hyperkatifiteia, watery discharge from eyes and nose (lacrimation and rhinorrhea), nausea, vomiting, diarrhea, abdominal pain, anorexia and fever. As noted above, one of the diagnostic tools developed regarding opioid withdrawal is the DSM-5. The DSM-5 specifies that for a subject to be diagnosed with opioid withdrawal, 3 of the following 9 symptoms must develop within minutes to several days of either cessation (or reduction) of opioid exposure, or the administration of an opioid antagonist or partial agonist. The DSM-5 symptoms are (1) dysphoric mood, (2) nausea, (3) muscle aches, (4) lacrimation or rhinorrhea, (5) pupillary dilation, piloerection or sweating, (6) diarrhea, (7) yawning, (8) fever and (9) insomnia. Accordingly, in some embodiments, the subject experiences at least 1, 2, 3, 4, 5, 6, 7, 8 or 9 of these DSM-5 symptoms and preferably administration of the compound of Formula (I) treats at least one of the symptoms experienced by the subject.
The severity of withdrawal symptoms will depend on the opioid causing the dependence, the dose and length of treatment or abuse, how rapidly opioid use is discontinued and the characteristics of the subject including age, sex, weight etc.
Accordingly, in some embodiments, the methods treat an opioid withdrawal symptom selected from the group consisting of tremors, shaking, hot or cold flashes, goosebumps, sweating, rapid breathing, elevated heart rate, elevated blood pressure, body aches, vomiting, diarrhea, fever, dysphoria, anxiety, restlessness, irritability, insomnia, yawning, hallucinations, hyperalgesia, hyperkatifiteia, and anorexia, or a combination thereof.
The salts and/or crystals of the invention may be administered by any suitable means, for example, orally, rectally, nasally, vaginally, topically (including buccal and sub-lingual), parenterally, such as by subcutaneous, intraperitoneal, intravenous, intramuscular, or intracisternal injection, inhalation, insufflation, infusion or implantation techniques (e.g., as sterile injectable aqueous or non-aqueous solutions or suspensions).
The salts and/or crystals of the invention may be provided as any suitable dosage form, including any of the medicaments and/or pharmaceutical compositions described herein.
Salts and/or crystals of the invention, may be administered in a dose of about 0.001, 0.005, 0.01, 0.05, 0.1, 0.15, 0.2, 0.5, 1, 2, 3, 5, 10, 15, 20, 25 or 30 mg/kg of the body weight of the subject. In some embodiments, the dose may be from any of these amounts to any other amount, such as from about 0.001 mg/kg to about 30 mg/kg, about 0.2 mg/kg to about 30 mg/kg or about 0.2 mg/kg to about 10 mg/kg. It will be understood, however, that the specific dose level and frequency of dosage for any particular subject may be varied and will depend upon a variety of factors including the activity of the specific compound employed, the metabolic stability and length of action of that compound, the age, body weight, general health, sex, diet, mode and time of administration, rate of excretion, drug combination, the severity of the particular condition, and the host undergoing therapy.
Salts and/or crystals of the invention may be administered in an “effective amount”, for example when an appropriate amount is included in a pharmaceutical composition. “Effective amount” is taken to mean an amount of a compound that will elicit a desired biological or medical response of a tissue, system, animal or human that is being sought by the researcher, veterinarian, medical doctor or other clinician administering the salts and/or crystals of the invention or a composition including the salts and/or crystals of the invention. In some embodiments, the effective amount may be a “therapeutically effective amount” wherein the amount of the salts and/or crystals of the invention is effective to treat the condition and/or symptom thereof that has manifested in the subject. In other embodiments, the effective amount may be a “prophylactically effective amount” wherein the amount of the salts and/or crystals of the invention is sufficient to prophylactically treat and/or prevent the onset of the condition and/or a symptom thereof or, if a symptom emerges, cause the severity of the condition and/or symptom thereof to be at a reduced level compared to the average severity of the condition and/or symptom thereof in a population of subjects not having received treatment with the compound of Formula (I) and/or a pharmaceutically acceptable salt and/or prodrug thereof.
The “effective amount” will be dependent on a number of factors, including the physical condition of the subject to be treated, the severity of symptoms, the formulation of the compound, and/or a professional assessment of the medical situation. The subject's weight and age may also be a factor for the person skilled in the art when determining the amount of salts and/or crystals of the invention that the subject should receive.
The phrases “administration of” and or “administering a” salt and/or crystal of the invention should be understood to mean providing the object active compound to a subject in need thereof.
As provided herein, beneficial or desired clinical results from the disclosed salt and/or crystal of the invention, include, without limitation, cessation of a symptom of the object disease, disorder or condition; alleviation of severity of a symptom of the object disease, disorder or condition; prevention of onset of a symptom of the object disease, disorder or condition; and/or managing a symptom of the object disease, disorder or condition for example preventing worsening of severity of a symptom or causing the symptom to reduce in severity or cease within a shorter than expected time. Either therapeutic or preventative measures may be achieved. Those in need of treatment include those already experiencing the object disease, disorder or condition as well as those in which the object disease, disorder or condition is to be prevented. By treatment is meant inhibiting or reducing an increase in symptoms of the object disease, disorder or condition when compared to the absence of treatment, and is not necessarily meant to imply complete cessation of the relevant condition.
Thus, generally, the term “treatment” (and variations thereof including “treating”) means affecting a subject, tissue or cell to obtain a desired pharmacological and/or physiological effect, including the beneficial or desired clinical results discussed above.
Also provided is a kit of parts, comprising in separate parts:
In any of the kits disclosed herein, the salts and/or crystals of the invention may be formulated as a pharmaceutical composition optionally together with a pharmaceutically acceptable carrier, diluent and/or excipient. The pharmaceutical compositions may be formulated for administration by any route disclosed herein including for oral, rectal, nasal, topical (including buccal and sub-lingual), parenteral administration (including intramuscular, intraperitoneal, sub-cutaneous and intravenous), or in a form suitable for administration by inhalation or insufflation.
XRPD analysis was carried out on a PANalytical X'pert pro with PIXcel detector (128 channels), scanning the samples between 3 and 35° 2θ. The material was gently ground to release any agglomerates and loaded onto a multi-well plate with Kapton or Mylar polymer film to support the sample. The multi-well plate was then placed into the diffractometer and analysed using Cu K radiation (α1 λ=1.54060 Å; α2=1.54443 Å; β=1.39225 Å; α1: α2 ratio=0.5) running in transmission mode (step size 0.0130° 20, step time 18.87 s) using 40 kV/40 mA generator settings. Data were visualized and images generated using the HighScore Plus 4.7 desktop application (PANalytical, 2017).
The presence of crystallinity (birefringence) was determined using an Olympus BX50 microscope, equipped with cross-polarising lenses and a Motic camera. Images were captured using Motic Images Plus 2.0. All images were recorded using the 20× objective, unless otherwise stated.
Approximately, 5 mg of material was weighed into an open aluminium pan and loaded into a simultaneous thermogravimetric/differential thermal analyser (TG/DTA) and held at room temperature. The sample was then heated at a rate of 10° C./min from 20° C. to 300° C. during which time the change in sample weight was recorded along with any differential thermal events (DTA). Nitrogen was used as the purge gas, at a flow rate of 300 cm3/min.
Approximately, 5 mg of material was weighed into an aluminium DSC pan and sealed nonhermetically with a pierced aluminium lid. The sample pan was then loaded into a Seiko DSC6200 (equipped with a cooler) cooled and held at 20° C. Once a stable heat-flow response was obtained, the sample and reference were heated to 250° C. at scan rate of 10° C./min and the resulting heat flow response monitored. Nitrogen was used as the purge gas, at a flow rate of 50 cm3/min.
NMR experiments were performed on a Bruker AVIIIHD spectrometer equipped with a DCH cryoprobe operating at 500.12 MHz for protons. Experiments were performed in deuterated DMSO and each sample was prepared to about 10 mM concentration.
Approximately, 10-20 mg of sample was placed into a mesh vapour sorption balance pan and loaded into a DVS Intrinsic dynamic vapour sorption balance by Surface Measurement Systems. The sample was subjected to a ramping profile from 40-90% relative humidity (RH) at 10% increments, maintaining the sample at each step until a stable weight had been achieved (dm/dt 0.004%, minimum step length 30 minutes, maximum step length 500 minutes) at 25° C. After completion of the sorption cycle, the sample was dried using the same procedure to 0% RH and then a second sorption cycle back to 40% RH. Two cycles were performed. The weight change during the sorption/desorption cycles were plotted, allowing for the hygroscopic nature of the sample to be determined. XRPD analysis was then carried out on any solid retained.
Approximately 10-20 mg of sample was placed into a mesh vapour sorption balance pan and loaded into an IGASorp Moisture Sorption Analyser balance by Hiden Analytical. The sample was subjected to a ramping profile from 40-90% relative humidity (RH) at 10% increments, maintaining the sample at each step until a stable weight had been achieved (98% step completion, minimum step length 30 minutes, maximum step length 60 minutes) at 25° C. After completion of the sorption cycle, the sample was dried using the same procedure to 0% RH, and finally taken back to the starting point of 40% RH. Two cycles were performed. The weight change during the sorption/desorption cycles were plotted, allowing for the hygroscopic nature of the sample to be determined.
1-Methyl-1,4,5,10-tetrahydropyrazolo[3,4-b][1, 5]benzodiazepine freebase was prepared for use in a primary salt screen (Example 2) using the following procedure:
The isolated 1-methyl-1,4,5,10-tetrahydropyrazolo[3,4-b][1,5]benzodiazepine freebase was characterized by TG/DTA, DSC, DVS with post-DVS, XRPD, HPLC, 1H NMR and LC-MS, as per the methods detailed above. The 1-methyl-1,4,5,10-tetrahydropyrazolo[3,4-b][1,5]benzodiazepine freebase was stored at 40° C./75% relative humidity (RH) for 1 week to determine the stability of the material at increased RH and to identify potential hydrate formation.
The following results were obtained during the preparation of the freebase:
The freebase of the compound of the invention was successfully produced and fully characterized. The material was found to be highly crystalline by XRPD. The material appeared anhydrous by TG analysis. DTA and DSC analysis confirmed a melting point of about 200° C. the freebase appeared hygroscopic by DVS with a mass increase of about 2.74% between 70 and 90% RH. Between 0 and 70% RH the material appears non-hygroscopic with a mass increase of about 0.11%. No change in form was observed by XRPD post-DVS analysis. The collected 1H NMR spectrum showed the expected connectivity of the structure provided. A high purity of 98.4% was confirmed b HPLC analysis. LC-MS showed a m/z of 201.3, corresponding to the expected mass of 200.24 g/mol. The freebase material stored at 40° C./75% RH for 1-week showed no changes in form by XRPD.
A salt screen was conducted on 1-methyl-1,4,5,10-tetrahydropyrazolo[3,4-b][1,5]benzodiazepine freebase using 5 solvent systems and 18 acid counterions (see Table 1).
The solvent systems used in this salt screen were (1) ethanol (EtOH); (2) tetrahydrofuran (THF); (3) isopropyl acetate; (4) acetone; and (5) 95% 2-propanol, 5% water (% v/v).
The following procedure was used:
During the primary salt screen of the 18 acid counterions tested, 16 produced potential solid salt forms of the compound. However, only the phosphate and L-tartrate salts possessed suitable properties based on thermal analysis and stability of the observed patterns at 40° C./75% RH.
Results of this salt screen are summarized in table 2.
From the above results, only the phosphate and L-tartrate salts possessed acceptable thermal stability and hygroscopicity.
During the primary polymorph screen, 16 potential salt forms were identified. Potential salt forms which appeared stable at 40° C./75% RH were analyzed by TG/DTA to identify salt forms with desirable thermal properties. TG/DTA analysis identified 4 potential salt forms with desirable properties; phosphate pattern 1 (phosphate form 1), L-tartrate pattern 1 (L-tartrate form 1), tosylate pattern 1 and tosylate pattern 3. Thermal analysis showed the phosphate form 1 and L-tartrate form 1 to be anhydrous.
This example describes protocols to prepare phosphoric acid addition salts of 1-methyl-1,4,5,10-tetrahydropyrazolo[3,4-b][1,5]benzodiazepine.
Experiment 3.1—Preparation from Freebase
The 1-methyl-1,4,5,10-tetrahydropyrazolo[3,4-b][1,5]benzodiazepine phosphate form 1 was prepared using the following procedure:
The dried material was fully characterised by TG/DTA, DSC, DVS with post-DVS, XRPD, HPLC, 1H NMR and LC-MS, as per the methods detailed above.
The isolated material appeared highly crystalline by XRPD (
Phosphate form 1 was successfully prepared by the above process. The isolated material appeared highly crystalline by XRPD and PLM. The observed crystals showed no defined morphology by PLM which may be due to the use of a stir-bar during preparation. TG analysis confirmed the material was anhydrous. DT and DSC analysis identified a melting point of about 200° C., consistent with the primary screen data. The material appeared slightly hygroscopic by DVS with a mass increase of about 0.4% at 90% RH (
The 1-methyl-1,4,5,10-tetrahydropyrazolo[3,4-b][1,5]benzodiazepine phosphate form 1 was prepared by reacting N-(1-methyl-1H-pyrazol-5-yl)-benzene-1,2-diamine with formaldehyde in the presence of phosphoric acid. N-(1-methyl-1H-pyrazol-5-yl)-benzene-1,2-diamine may be prepared as described in Katte, TA; Reekie, TA; Jorgensen, WT; and Kassiou, M; J. Org. Chem. 2016, 81(11), 4883-4889, which is entirely incorporated by reference including all supporting information.
N-(1-methyl-1H-pyrazol-5-yl)-benzene-1,2-diamine was recrystallised from ethyl acetate and heptane prior to inclusion in the following reaction. The recrystalised N-(1-methyl-1H-pyrazol-5-yl)-benzene-1,2-diamine had a purity of 99.4% area assessed by HPLC using conventional separation techniques. It will be appreciated that recrystalisation is not essential for successful completion of the following reaction with formaldehyde.
A 1 litre flask was fitted with a condenser, pressure equalising addition funnel, nitrogen inlet and magnetic stirrer bar and purged with nitrogen. Recrystalised N-(1-methyl-1H-pyrazol-5-yl)-benzene-1,2-diamine (37.2 g) was added to the flask, followed by a mixture of acetonitrile (184.6 g) and water (145.0 g) which had been previously degassed. The mixture was heated to 30° C., and shortly after 2.9 g of a mixture of phosphoric acid (23.1 g) and water (26.3 g) added, washing in with water (14.9 mL). After 10 minutes, 37% aqueous formaldehyde (15.9 g) was added over 25 minutes maintaining the temperature between 36° C. and 43° C. The solution was washed in with water (14.9 mL) within 10 minutes. Seventy (70) minutes after formaldehyde addition began, the mixture was filtered into a nitrogen purged receiver, and the reaction flask washed out with acetonitrile (7.9 mL), the wash being passed through the filter. The liquor flask was purged with nitrogen, and reheated to 34° C. The remaining phosphoric acid was diluted further using acetonitrile (49.2 g) and added over 35 minutes at 34° C. to 39° C., washing in with acetonitrile (5.8 g). The mixture was stirred for 5 minutes, then cooled to 19° C. over 2 hours, then cooled in an ice bath for 105 minutes. At 6° C., the mixture was filtered, the cake washed with approximately two thirds of a mix of acetonitrile (26.4 g) and water (22.8 g), then the cake compressed. The filter cake was washed with the remaining third of the acetonitrile/water mixture, dried for 2 minutes on the sinter with a nitrogen blanket. The pale pink solid (66.9 g) was transferred to an oven and dried at 40° C. for 115 hours to provide Phosphate Form 1 (Yield 51.0 g, 86.5%).
Lyophilized material of the phosphoric acid of the compound of the invention was prepared using the following procedure:
Selected solvent was added in 50 μL aliquots to approximately 10 mg of poorly crystalline phosphate pattern (obtained from above lyophilisation). Between each addition, the mixture was checked for dissolution and where no dissolution was apparent, the mixture was heated to about 40° C. and checked again. This procedure was continued until dissolution was observed or until 2 mL of solvent had been added. Samples in which dissolution was observed were stored uncapped at ambient temperature to allow evaporation. Samples in which dissolution was not observed were filtered via centrifugation. All observed solids were analyzed by XRPD. The solvent systems used during the solvent solubility screen are detailed in Table 3.
During the solvent solubility screen the following results were obtained:
Phosphate form 1 was produced in the majority of the above samples, with the exception of chloroform (traces of pattern 2 were observed), dimethylsulfoxide (no solid), ethyl acetate (pattern 2), ethyl formate (mix of form 1 and pattern 2), isopropyl acetate (mix of form 1 and pattern 2), methylethyl ketone (mix of form 1 and pattern 2), methylisobutyl ketone (pattern 2), N-methyl pyrolidone (no solid), tert-butylmethylether (pattern 2) and water (no solid).
The following procedure was used during the polymorph screen maturation experiments:
Phosphate form 1 was observed predominantly in the maturation experiments. A mixture of form 1 and pattern 2 was observed from ethyl formate. Phosphate pattern 2 was observed from tertbutylmethyl ether only.
Phosphate pattern 2 material isolated from tert-butyl methyl ether was collected and dried under vacuum at 40° C. for about 2 hours. The dried material was collected and analyzed by TG/DTA.
The following observations and results were obtained during the characterization of phosphate pattern 2:
The XRPD multi-well plate containing the phosphate pattern 2 material observed in the solvent solubility screen was collected and stored in a stability chamber at 40° C./75% RH for about 16 hours. The samples were analyzed by XRPD.
The phosphate pattern 2 was determined to have lower melting point than form 1, indicating the material is less stable. Upon storage at 40° C./75% RH phosphate pattern 2 material completely converted to form 1, confirming pattern 2 to be a metastable form of the phosphate salt.
The 1-methyl-1,4,5,10-tetrahydropyrazolo[3,4-b][1,5]benzodiazepine L-tartrate salt was prepared using the following procedure:
The dried material was fully characterised by TG/DTA, DSC, DVS with post-DVS, XRPD, HPLC, 1H NMR and LC-MS, as per the methods detailed above.
The isolated material appeared highly crystalline by XRPD (
The L-tartrate form 1 salt was successfully prepared by the above process. The isolated material appeared highly crystalline by XRPD and PLM. The observed crystals showed no defined morphology by PLM which may be due to the use of a stir bar during preparation. Agglomeration of particles was observed in the PLM analysis. TG analysis confirmed the material was anhydrous. DT and DSC analysis identified a melting point of about 181° C., consistent with the primary screen data. The material appeared slightly hygroscopic by DVS with a mass increase of about 0.6% at 90% RH. No evidence of form changes or hydrate formation was observed by XRPD post-DVS analysis. The collected 1H NMR spectrum showed the expected connectivity with the structure provided. Approximately 1 equivalent of L-tartaric acid was observed in the 1H NMR spectrum. Small impurities were observed in the 1H NMR spectrum which were later identified in the L-tartaric acid input material. HPLC analysis confirmed a high purity of 99.0% (by area %), showing a small uplift in purity from the freebase input material.
One-week stability studies were conducted on the freebase (example 1), phosphate form 1 (example 3) and L-tartrate form 1 (example 5) salts using the following procedures:
During the 1-week stability studies, no changes in form were identified in the phosphate form 1 and L-tartrate form 1 salts by XRPD. HPLC analysis did not identify any significant changes in purity in the salt samples.
The freebase samples stored at 40° C./75% RH and 80° C. showed no changes in form or purity. An additional peak was observed in the XRPD diffractogram of the freebase sample stored under ambient conditions, indicating a potential change in form or degradation. An additional XRPD pattern collected after 2 weeks showed further changes in diffraction pattern. HPLC analysis did not show any significant changes in purity. The data indicates that the freebase may be unstable under ambient light, but otherwise stable under thermal conditions.
The following experiments were conducted to assess the likelihood of salt disproportionation of the phosphate form 1 and L-tartrate form 1 salts in deionised water and the potential for hydrate formation using solvent/water mixtures with various water activities.
The following procedure was used during the salt disproportionation studies:
The following procedure was used during the hydration studies:
The following results were obtained during the salt disproportionation studies of phosphate form 1 (example 3) and L-tartrate form 1 (example 5) salts:
The following results were obtained during the hydration studies of phosphate form 1 (example 3) and L-tartrate form 1 (example 5) salts:
No evidence of hydrate formation or salt disproportionation was observed in the phosphate form 1 and L-tartrate salts.
The thermodynamic solubility of freebase (example 1), phosphate form 1 (example 3) and L-tartrate form 1 (example 5) were assessed in phosphate buffered saline (PBS) buffer at pH 7.4 according to the following procedure:
The following results (summarised in Table 6) were obtained during the thermodynamic solubility assessment of freebase (example 1), phosphate form 1 (example 3) and L-tartrate form 1 (example 5):
During the thermodynamic solubility screen, the KNX-100 freebase was found to have the lowest solubility in PBS buffer at pH 7.4, with a solubility of 0.3 mg/mL. Higher solubilities were identified in the salt forms. KNX-100 L-tartrate form 1 had a solubility of 6.3 mg/mL and the KNX-100 phosphate pattern 1 was found to have the highest solubility of 7.4 mg/mL.
This Example describes pharmacokinetic experiments in male Sprague Dawley rats. The Example show that oral administration of the compound of the invention (CMPD1) as phosphate form 1 leads to the same exposure profile as the compound of the invention dosed in dihydrochloride salt form, whether the drug is administered using a saline or methocel vehicle.
N=3 rats were run in each of the four conditions:
On the day of dosing the solid compounds were dissolved in either Saline (0.9%) or 0.5% hydroxypropyl methylcellulose (Methocel E3 Premium LV) in Milli-Q water. The formulations were then thoroughly vortexed to produce colourless solutions.
Overnight-fasted rats with adlibitum access to water were administered their dose of the various forms of the compound of the invention via oral gavage (PO) at a dose volume of 3 ml/kg and a dose of 5 mg/kg freebase equivalent. Food access was re-instated 4 hours (h) post-dose. Samples of arterial blood were collected up to 24 h post-dose. After collection, samples were centrifuged, plasma was removed and stored frozen at −80° C. before being analysed by LC-MS. Urine samples were collected at pre, 0-4 h, 4-7 h and 7-24 h post-dose and were analysed by LC-MS following extraction.
No adverse reactions or compound-related side effects were observed in any rats during the 24 h sampling period after dosing.
The mean plasma concentration of the compound of the invention versus time profiles of following oral administration of the di-hydrochloride salt and phosphate form 1 in each formulation are shown in
The plasma concentration versus time profiles of the compound of the invention were closely comparable for all four treatment groups. This was reflected in the similar dose-normalised Cmax and AUC0-inf values, suggesting that neither the salt form nor formulation vehicle had any substantial impact on the exposure of the subject to the compound of the invention.
This Example describes experiments in a C57BL/6 mouse model of opioid withdrawal (naloxone precipitated withdrawal following oxycodone administration) and the potential of the compound of the invention in two different salt forms, administered at the same freebase equivalent dose, to treat withdrawal symptoms. This experiment confirms that a substantially similar biological activity is achieved for phosphate form 1 as for previously described forms of the compound of the invention.
Two cohorts of adult male C57BL/6 mice were run assessing the effects of 7.3 mg/kg freebase equivalent (FBE) IP of the compound of the invention in the dihydrochloride salt form (CMPD1-2HCL) and the phosphate form 1 (CMPD1-PO4) on naloxone precipitated withdrawal-induced jumping.
The first cohort of mice (N=30) were split into the following conditions:
The second cohort of mice (N=24) were split into the following conditions:
Mice in the oxycodone conditions received i.p. injections of oxycodone for 5 days according to the schedule and doses set out in Table 8. The morning and afternoon doses were separated by 7 h. Mice in the vehicle condition received injections of vehicle saline instead of oxycodone. One-hour-and-forty-five minutes after the morning injection on day 5, mice were administered their i.p. dose of the compound of the invention. Fifteen minutes later they received an i.p. injection of 10 mg/kg naloxone (oxycodone groups) or saline (vehicle group), and proceeded immediately to testing.
Testing involved placing mice individually into a 20 (I)×20 (w)×30 (h) cm arena for 30 min. Sessions were captured via a side view high speed (120 fps), high resolution (4K) camera. Number of jumps were scored from the videos by an experienced experimenter blind to conditions.
Data were analysed by SPSS using oneway ANOVA and planned contrasts.
Data for jumping are shown in
The overall ANOVA assessing jumping was significant [F3,50=21.52, p<0.0001]. Planned contrasts revealed mice undergoing oxycodone withdrawal jumped significantly more times during the test session [VEH_VEH vs OXY_VEH, p<0.0001].
7.3 mg/kg FBE compound of the invention in both the phosphate (CMPD1-PO4) and 2HCl (CMPD1-2HCL) salt form significantly inhibited oxycodone withdrawal-induced jumping at all doses tested [OXY_VEH vs: OXY_CMPD1-PO4, p<0.01; OXY_CMPD1-2HCL, p<0.001]. Moreover, the results following dosing FBE of the two salt forms did not differ significantly from each other in withdrawal-induced jumping [OXY_CMPD1-PO4 vs OXY_2HCL, p=0.901] (
Number | Date | Country | Kind |
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2020903196 | Sep 2020 | AU | national |
Filing Document | Filing Date | Country | Kind |
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PCT/AU2021/051033 | 9/7/2021 | WO |