WO2020174238A1 - Prochlorperazine for the treatment of pitt-hopkins syndrome - Google Patents

Abstract

The present invention relates to prochlorperazine or a pharmaceutically acceptable sale thereof, for use in the treatment of Pitt-Hopkins syndrome.

Description

PROCHLORPERAZINE FOR THE TREATMENT OF

PITT-HOPKINS SYNDROME

Field of the invention

This invention relates to the treatment of Pitt-Hopkins syndrome (PTHS). Background of the invention

Pitt-Hopkins syndrome is a rare, genetic neurological disorder caused by a molecular variant of TCF4 which is involved in embryologic neuronal differentiation.

PTHS is characterised by distinctive facial features (syndromic facies), developmental delay (psychomotor delay), intellectual disability, eariy-onset myopia, seizures, constipation and breathing abnormalities (hyperventilation-apneic spells i.e. recurrent episodes where they breathe very fast, often followed by episodes where they struggle to breathe or momentarily stop breathing) and low muscle tone (hypotonia). Further symptoms include repetitive nonfunctional hand movements and behavioural abnormalities such as hyperactivity and anxiety. Many affected individuals meet criteria for autism spectrum disorder.

PTHS is caused by a pathogenic variant of the TCF4 gene found on chromosome 18q21.2. The syndrome was first described in 1978 in two unrelated individuals who shared similar characteristics of dysmorphic facial features, developmental delay, dubbed fingers, and an abnormal breathing pattern. It was presumed to be an autosomal recessive disorder until 2007 when the TCF4 gene, (MIM #602272) was identified, supporting an autosomal dominant inheritance pattern secondary to haploinsuffiency of TCF4. Transcription factor 4, the protein product of TCF4, is a basic he!ix-loop-helix E-protein believed to be involved in early brain development and neuronal differentiation.

At present there is no effective therapy to treat PTHS. There is a need for therapies for the treatment of PTHS.

Prochlorperazine is a phenothiazine with antiemetic, antipsychotic, antihistaminic and anticholinergic activities. Prochlorperazine is a dopamine D2- receptor antagonist. Prochlorperazine is used in the treatment of nausea, vomiting and vertigo.

Prochlorperazine has the systematic name 2-chloro-10-[3-(4-methylpiperazin- 1 -yl)propyl]phenothiazine. Prochlorperazine is marketed under the trade names Ametil®, Antinaus®, Antinaus®, Buccastem®, Bukatel®, Chlormeprazine®, Ch!oropernazine®, Compazine®, Compro®, Daolin®, Dhaperazine®, Emedrotec®, Emetirai®, Eminorm®, Lotamin®, Mitil®, Morma®I, Nautisol®, Novamin®, Novomit®, Proazine®, Procalm®, Prochiorperazin®, Prochlorperazine®, Prochlorperazine®,

Prochlorperazinum®, Prochlozine®, Proclorperazina®, Promat®, Promin®, Promtii®, Roumin®, Scripto-metic®, Seratil®, Stemetii®, Steremai®, Vergon®, Vestii®, and Volimin®.

Summary of the invention

The present invention is prochlorperazine, or a pharmaceutically acceptable salt thereof, for use in the treatment of PTHS. As will be evident from the in vivo data presented below, prochlorperazine is effective in treating PTHS. Chronic treatment with prochlorperazine significantly improved the Tcf4+/- mouse phenotype, rescuing fear conditioning, open field, nesting, seif-grooming, sociability and test of force. This is evidence that prochlorperazine is useful in the therapy of PTHS.

A first aspect of the invention is prochlorperazine, or a pharmaceutically acceptable salt thereof, for use in the treatment of Pitt-Hopkins syndrome.

Description of the figures

Figure 1 shows the results from prochlorperazine in vivo testing for fear conditioning (learning and memory).

Figure 2 shows the results from prochlorperazine in vivo testing for open field (hyperactivity).

Figure 3 shows the results from prochlorperazine in vivo testing for nesting (tests of daily living).

Figure 4 shows the results from prochlorperazine in vivo testing for self- grooming (stereotypy i.e. repetitive movement).

Figure 5 shows the results from prochlorperazine in vivo testing for sociability.

Figure 6 shows the results from prochlorperazine in vivo testing test of force (strength).

Figure 7 shows the results of dose response experiments in 5 behavioural tests. In the Figures, ‘ns’ means no significant difference from Wild Type control group. This indicates that the drug-treated Knock Out is behaving like the untreated or treated Wild Type group in the behavioural test, meaning the compound was significantly effective in ameliorating the syndrome’s phenotype.

Detailed description

There are a number of different manifestations and symptoms in patients with Pitt-Hopkins syndrome. These include: hyperactivity and repetitive behaviour including repetitive nonfunctional hand movements; intellectual impairment, such as difficulties with learning and memory e.g. difficulties with cognitive, executive and language performance, short-term memory, executive function, visual memory, visual- spatial relationships and spatial working memory; stereotypy; social anxiety (i.e difficulties in social interaction); low muscle tone (hypotonia); constipation, sleep disturbances, seizures; irregular or abnormal breathing patterns and severe nearsightedness (myopia).

in the present invention, and as demonstrated by the below in vivo data, prochlorperazine is used to treat one or more of the above symptoms, and is therefore an effective treatment of PTHS Preferably, prochlorperazine is used for the treatment of PTHS, wherein the patient is exhibiting typical symptoms of the syndrome including hyperactivity; social anxiety; intellectual impairment, specifically difficulties with learning and memory; stereotypy; and hypotonia.

The term “intellectual impairment” has its normal meaning in the art. It encompasses impairment in learning and memory. Learning impairment may also be called intellectual disability. It encompasses cognitive impairment, delay or limitations in intellectual functions, such as reasoning. Memory impairment refers to an inability to retain information either short-term or long-term. It may include difficulties with cognitive, executive and language performance, executive function and visual memory. It may also include difficulties with working memory, also called short-term memory (i.e. the temporary storage of information while processing the same or other information) and difficulties with phonological memory (or verbal working memory). This symptom was tested in mice under“fear conditioning” (see in vivo data below).

The term“hyperactivity” has its normal meaning in the art. Hyperactivity may include having very short attention spans, hypersensitivity to visual, auditory, tactile, and olfactory stimuli, distractibility, impuisiveness, restlessness and/or over-activity. This symptom was tested in mice under“open field” (see in vivo data below).

The term“test of daily living” has its normal meaning in the art it may also mean ability to perform the things normal to a species including any daily activity we perform such as bedding, feeding etc. This symptom was tested in mice under “nesting” (see in vivo data below).

The term“stereotypy” has its normal meaning in the art. it may also be termed as repetitive movements or repetitive behaviour. This symptom was tested in mice under“seif-grooming” (see in vivo data below).

The term“social anxiety” has its normal meaning in the art. It may also be termed as difficulties in social interaction or low sociability. Social anxiety may include having poor eye contact, gaze aversion, prolonged time to commence social interaction, social avoidance or withdrawal and challenges forming peer relationships. This symptom was tested in mice under“sociability” (see in vivo data below).

The term“hypotonia” has its normal means in the art. This symptom was test in mice under "test of force”. The term“force” has its normal meaning in the art. it may also mean the strength or energy put into an action.

As used herein, a pharmaceutically acceptable salt is a salt with a pharmaceutically acceptable acid or base. Pharmaceutically acceptable acids include both inorganic acids such as hydrochloric, sulphuric, phosphoric, diphosphoric, hydrobromic or nitric acid and organic acids such as citric, fumaric, maleic, malic, ascorbic, succinic, tartaric, benzoic, acetic, methanesuifonic, ethanesulfonic, salicylic, stearic, benzenesuifonic orp-toluenesu!fonic acid. Pharmaceutically acceptable bases include alkali metal (e.g. sodium or potassium) and alkali earth metal (e.g. calcium or magnesium) hydroxides and organic bases such as alkyl amines, aryl amines or heterocyclic amines.

In the present invention, prochlorperazine may be administered in a variety of dosage forms in one embodiment, prochlorperazine may be formulated in a format suitable for oral, rectal, parenteral, intranasal or transdermai administration or administration by inhalation or by suppository.

Prochlorperazine may be administered orally, for example as tablets, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules. Preferably, prochlorperazine is formulated such that it is suitable for oral administration, for example tablets and capsules.

Prochlorperazine may also be administered parenterally, whether subcutaneously, intravenously, intramuscularly, intrasternally, transdermally or by infusion techniques. Prochlorperazine may also be administered as suppositories.

Prochlorperazine may also be administered by inhalation. An advantage of inhaled medications is their direct delivery to the area of rich blood supply in comparison to many medications taken by oral route. Thus, the absorption is very rapid as the alveoli have an enormous surface area and rich blood supply and first pass metabolism is bypassed.

The present invention also provides an inhalation device containing prochlorperazine. Typically said device is a metered dose inhaler (MDI), which contains a pharmaceutically acceptable chemical propellant to push the medication out of the inhaler.

Prochlorperazine may also be administered by intranasal administration. The nasal cavity's highly permeable tissue is very receptive to medication and absorbs it quickly and efficiently. Nasal drug delivery is less painful and invasive than injections, generating less anxiety among patients. By this method absorption is very rapid and first pass metabolism is usually bypassed, thus reducing inter-patient variability. Further, the present invention also provides an intranasal device containing prochlorperazine.

Prochlorperazine may also be administered by iransdermal administration. For topical delivery, transdermal and transmucosal patches, creams, ointments, jellies, solutions or suspensions may be employed. The present invention therefore also provides a iransdermai patch containing a prochlorperazine.

Prochlorperazine may also be administered by sublingual administration. The present invention therefore also provides a sub-lingual tablet comprising prochlorperazine.

Prochlorperazine may also be formulated with an agent which reduces degradation of the substance by processes other than the normal metabolism of the patient, such as anti-bacterial agents, or inhibitors of protease enzymes which might be the present in the patient or in eommensura! or parasite organisms living on or within the patient, and which are capable of degrading the compound.

Liquid dispersions for oral administration may be syrups, emulsions and suspensions.

Suspensions and emulsions may contain as carrier, for example a natural gum, agar, sodium alginate, pectin, metby!cei!uiose, carboxymetby!cei!uiose, or polyvinyl alcohol. The suspension or solutions for intramuscular injections may contain, together with the active compound, a pharmaceutically acceptable carrier, e.g. sterile water, olive oil, ethyl oieate, glycols, e.g. propylene glycol, and if desired, a suitable amount of lidocaine hydrochloride.

Solutions for injection or infusion may contain as carrier, for example, sterile water or preferably they may be in the form of sterile, aqueous, isotonic saline solutions.

In an embodiment of the invention, prochlorperazine is administered in an effective amount to treat the symptoms of Pitt-Hopkins syndrome. An effective dose will be apparent to one skilled in the art, and is dependent on a number of factors including age, sex, weight, which the medical practitioner will be capable of determining.

In a preferred embodiment, prochlorperazine is administered in doses of 1 to 60 mg, 5 mg to 60 mg, more preferably 10 mg to 60 mg, most preferably 20 mg to 40 mg. The lower limit for a dose is preferably 1 mg, 2 mg, 3 mg, 4 mg or 5 mg, 10 mg, 15 mg, 20 mg or 25 mg. The upper limit for a dose is preferably 60 mg, 55 mg, 50 mg, 45 mg, 40 mg, 35 mg or 30 mg. Any of the aforementioned lower or upper limits of the ranges may be combined with each other, and are herein disclosed. Preferably the dose is 1 to 20 mg or 20 mg to 40 mg.

Any of the above doses may be administered once a day, twice a day, three times a day or four times a day.

In an embodiment of the invention, prochlorperazine is administered at least once a day. Preferably it is administered as a single daily dose. Preferably the single daily dose is of 5 mg to 60 mg, more preferably 20 to 40 mg. Preferably it is 40 mg, 30 mg or 20 mg. In an embodiment of the invention, prochlorperazine is administered twice daily. Preferably each dose is 1 mg to 30 mg or 5 mg to 30 mg, with a total daily dosage of 2 mg to 60 mg or 10 mg to 60 mg. Preferably, each dose is 2 mg or 20 mg.

Alternatively, it may be administered three times per day. Preferably each dose is 1 mg to 30 mg or 5 mg to 20 mg. More preferably, each does is 10 to 15 mg.

Alternatively, it may be administered four times per day. Preferably each dose is 5 mg to 15 mg. Preferably, each does is 10 mg.

Preferably, the dosage regime is such that the total daily dosage of prochlorperazine does not exceed 60 mg.

It will be appreciated that a lower dose may be needed in a paediatric patient. For example, a dose of about 2 mg may be appropriate in a paediatric patient.

In order to treat Pitt-Hopkins syndrome, prochlorperazine is used in a chronic dosage regime i.e. chronic, long-term treatment.

The present invention also relates to use of prochlorperazine, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for use in the treatment of Pitt-Hopkins syndrome. This embodiment of the invention may have any of the preferred features described above.

The present invention also relates to a method of treating Pitt-Hopkins syndrome comprising administering the patient with prochlorperazine or a pharmaceutically acceptable salt thereof. This embodiment of the invention may have any of the preferred features described above. The method of administration may be according to any of the routes described above.

For the avoidance of doubt, the present invention also embraces prodrugs which react in vivo to give a compound of the present invention.

The following studies illustrate the invention.

Study 1

Pitt-Hopkins syndrome (PTHS) is characterized by cognitive dysfunction, wide mouth and distinctive facial features, and intermittent hyperventilation followed by apnoea (Zwei er et al., 2007). The cognitive dysfunction associated with the loss of one copy of the TCF4 gene, termed haplo-lnsufficiency, in humans leads to Pitt-Hopkins syndrome, an autism-related disorder associated with pronounced learning deficits. Pitt-Hopkins syndrome (PTHS) is a genetic neurodeve!opmental disorder associated with transcription factor TCF4 mutations/deletions. TCF4 may also be linked to schizophrenia, suggesting that the precise pathogenic mutations are relevant to cellular, synaptic, and behavioural consequences.

The Tcf4 +/- mouse model has been developed to mimic PTHS and therefore assess potential treatments. The mice have deficits in hippocampus-dependent learning and memory, spatial working memory, sociability, stereotypy, hyperactivity and daily living paradigms. The Tcf4 +/- mice also demonstrate hindiimb grip strength deficits.

Animals

Mice were purchased from The Jackson Laboratory and maintained on a C57BL/8 background. They were raised on a 12:12 lightdark cycle with ad libitum access to food and water. Controls consisted of TCF4 +/+. Ail mice used were heterozygous for the Tcf4 mutation because homozygous mutations result in embryonic /post-partum lethality.

Assay design

Experiments were randomised and blind to the genotype and treatment during ail testing and data analysis. Separate investigators prepare and coded dosing solutions, allocate the mice to the study treatment groups, dosed the animals, and collect the behavioural data.

Treatment Groups

There were four treatment groups per compound in the study with 10 male mice used per treatment group (ail at 14 weeks of age): Group 1 : wild-type littermate mice treated with vehicle (WT + veh); Group 2: Tcf4 +/- KO mice treated with vehicle (Tcf4 +/- + veh), Group 3: WT + drug; and Group 4: Tcf4 +/- + drug.

Behavioural and Strength Tests

This included:

1. Fear conditioning (a test of learning and memory);

2. Open field (a measure of hyperactivity)

3. Nesting (a test of daily living)

4. Self-grooming (an assessment of stereotypy). 5. Sociability/Partition test (an assessment of social anxiety)

6 Test of Force (a test of hind limb strength).

The term learning and memory” has its norma! meaning in the art. It may also be called memory impairment it refers to an inability to retain information either short term or long-term it may include difficulties with cognitive, executive and language performance, executive function and visual memory it may also include difficulties with working memory, also called short-term memory (i.e. the temporary storage of information while processing the same or other information) and difficulties with phonological memory (or verbal working memory).

The term“hyperactivity” has its normal meaning in the art. Hyperactivity may include having very short attention spans, hypersensitivity to visual, auditory, tactile, and olfactory stimuli, distractibility, impulsiveness, restlessness and/or over-activity.

The term“test of daily living” has its norma! meaning in the art. It may also mean ability to perform the things norma! to a species including any daily activity we perform such as bedding, feeding etc.

The term‘seif-grooming’ has its normal meaning in the art. It may also mean self-cleaning and maintaining normal appearance of skin, hair, fur etc. Carried to excess via persistent repetition of an act it may be referred to as‘sterotypy’.

The term“social anxiety” has its norma! meaning in the art. It may also be termed as difficulties in social interaction or low sociability. Social anxiety may include having poor eye contact, gaze aversion, prolonged time to commence social interaction, social avoidance or withdrawal and challenges forming peer relationships.

The term“force” has its normal meaning in the art. It may also mean the strength or energy put into an action.

1. Fear Conditioning

Tcf4 +/__ mice show reduced freezing in the context of clued fear conditioning test indicating hippocampus-dependent memory and learning deficiency. The dependent measure used in contextual fear conditioning was a freezing response following a pairing of an unconditioned stimulus (foot shock), with a conditioned stimulus, a particular context. Freezing is a species-specific response to fear, which has been defined as“absence of movement except for respiration”. This may last for seconds to minutes depending on the strength of the aversive stimulus, the number of presentations, and the degree of learning achieved by the subject. Testing involved placing the animal in a novel environment (dark chamber), providing an aversive stimulus (a 1 -sec electric shock, 0.2 mA, to the paws), and then removing it.

2. Open Field

The open-field apparatus was used to test hyperactivity Tcf4÷/__ mice show hyperactivity when compared to WT littermates in distance travelled per minute during a 30 minutes trial Open Field. The apparatus was a grey PVC-enciosed arena 50x9x30 cm divided into a 10x10cm grid. Mice were brought to the experimental room 5-~20min before testing. A mouse was placed info a corner square facing the corner and observed for 3min. The number of squares entered by the whole body (locomotor activity) was counted. The movement of the mouse around the field was recorded with a video tracking device for 3min (version NT4.0, Viewpoint).

3. Nesting

The test was performed in individual cages. Normal bedding covered the floor to a depth of 0.5 cm. Each cage was supplied with a“Nestlet,” a 5 cm square of pressed cotton batting (Ancare). Mice were placed individually info the nesting cages 1 hr. before the dark phase, and the results were assessed the next morning. Nest building was scored on a 5 point scale.

Score 1 : The Nestlet was largely untouched (>90% intact).

Score 2: The Nestlet was partially torn up (50-90% remaining intact).

Score 3: The Nestlet was mostly shredded but often there was no identifiable nest site: < 50% of the Nestlet

Score 4: An identifiable, but flat nest < 90% of the Nestlet was torn up, the material was gathered into a flat nest with wails higher than the mouse height curled up on its side) on less than 50% of its circumference.

Score 5: A (near) perfect nest: >90% of the Nestlet was torn up, the nest was a crater, with walls higher than mouse body height on more than 50% of its circumference.

4. Self-grooming

Tcf4 +/_ mice groom themselves significantly more than WT mice indicating higher leveisof stereotype behaviour than control mice. Usually in a sitting position, the mouse will lick its fur, groom with the forepaws, or scratch with any limb. Often the mouse will mix all of these grooming behaviors. Grooming typically follows a sequence of four behaviors:

Elliptical Stroke: Elliptical asymmetric movements of the forepaws over the nose and muzzle, alternating the major and minor paw.

Unilateral Stroke: Alternating strokes of the forepaw across the vibrissae and the eye.

Bilateral Stroke: Large symmetric bilateral strokes of the forepaws that begin behind the ears and pass over the whole face.

Body Licking: Licking of the whole body, typically beginning rostrally and working caudally to the tail.

5. Partition test

In the three-chambered sociability task, a subject mouse was evaluated for its exploration of a novel social stimulus (novel mouse). The three-chambered social approach task monitors direct social approach behaviours when a subject mouse is presented with the choice of spending time with either a novel mouse or an empty cup. Sociability is defined as the subject mouse spending more time in the chamber containing the mouse than in the empty chamber. Preference for social novelty is defined as spending more time in the chamber with the novel mouse. The apparatus is a rectangular three-chamber box, where each chamber measures 20 cm (length) x 40.5 cm (width) x 22 cm (height). Dividing walls are made from dear perplex, with small openings (10 cm width x 5 cm height) that allow access into each chamber. The three chamber task was lit from below (10 lux). The mice were allowed to freely explore the three-chamber apparatus over three 10 min trials. During the trial one wire cup was placed upside down in one of the side chambers and a novel mouse was placed under another wire cup in the other side chamber (novel mouse stimulus), leaving the middle chamber empty. The location of the novel mouse across trials was counterbalanced to minimize any potential confound due to a preference for chamber location. The time spent exploring the novel mice was scored as exploration ratio.

8. Test of Force

Tcf4+/- mice show a significant deficit in hind!imb strength (not forelimb) when compared with WT mice. Neuromuscular function of the hindlimbs was tested with a grip strength meter (San Diego instruments). Mice were scruffed by the back of the neck, held by the tail, and lifted into an upright position. Then, the mice were lowered toward the apparatus, allowed to grasp the smooth metal grid (hindiimbs only), and pulled backwards in the horizontal plane. The force applied to the grid at the moment the grasp was released was recorded.

Statistical Analysis of Behavioural Data

Data were analysed by two-way analysis of variance (ANOVA) followed by posttest comparisons where appropriate using Tukey's Multiple Comparison Test. Data are represented as the mean and standard error of the mean (SEM).

Conclusion

As is evident from the data above, chronic treatment of Tcf4+/- mice with prochlorperazine significantly improved learning and memory; hyperactivity; tests of daily living; stereotypy; social anxiety and test of force. As the mouse model mimics PTHS, this is evidence that prochlorperazine has a therapeutic effect for PTHS.

Study 2

Dose response experiments were conducted following the protocol described above. The results are shown in Figure 7 Prochlorperazine rescued all the behavioral tests at 1.5 mg/kg. Prochlorperazine rescued the following behavioral tests at 0.8 mg/kg: Open Field & Nesting.

Claims

Claims

1. Prochlorperazine, or a pharmaceutically acceptable salt thereof, for use In the treatment of Pitt-Hopkins syndrome.

2. Prochlorperazine for use according to claim 1 , wherein the subject of treatment is human.

3. Prochlorperazine for use according to any of claims 1 or 2, wherein the dose of prochlorperazine is 1 to 60 mg, preferably 5 mg to 60 mg, preferably 10 mg to 50 mg, most preferably 20 mg to 40 mg.

4. Prochlorperazine for use according to any preceding claim, wherein administration is by a single daily dose.

5. Prochlorperazine for use according to claim 4, wherein the single daily dose is 20 mg to 40 mg.

6. Prochlorperazine for use according to any of claims 1 to 3, where administration is by a dose twice per day.

7. Prochlorperazine for use according to claim 6, wherein the dose is 1 mg to 30 mg, preferably 5 g to 30 mg, more preferably 20 mg.

8. Prochlorperazine for use according to any preceding claim, to be administered orally.

9. Prochlorperazine for use according to any of claims 1 to 7, to be administered by parenteral, transdermai, sublingual, rectal or inhaled administration.

10. Prochlorperazine for use according to any preceding claim, wherein the patient is exhibiting signs of intellectual impairment, hyperactivity, stereotypy, social anxiety and/or hypotonia.

11. A method of treating Pitt-Hopkins syndrome comprising administering the patient with prochlorperazine or a pharmaceutically acceptable salt thereof.

12. The method according to claim 11 , having any of the additional features of claims 2 to 9.