DESC:DESCRIPTION OF THE FIGURES
Figure 1: Nigral TH+ve cell counts (operated side).
Figure 2: Striatal TH OD (operated vs non-operated sides).
Figure 3: Striatal dopamine levels: Absolute values, operated vs non-operated
5 sides.
DETAILED DESCRIPTION OF THE INVENTION
In one aspect, the present invention provides a method of treating or
preventing synucleinopathies in a human subject comprising administering a
therapeutically effective amount of the compound of Formula 1
N
NH
NH
Cl
CH3
H3C
O
O
Formula 1 10
or its pharmaceutically acceptable salt thereof.
In another aspect the present invention provides a method of treating or
preventing synucleinopathies in a human subject comprising administering
compound of Formula 1
N
NH
NH
Cl
CH3
H3C
O
O
Formula 1 15
or its pharmaceutically acceptable salt thereof at a dose ranging from 0.1 mg
to 1000 mg per day.
In an embodiment, the present invention provides a method of treating
synucleinopathies in a human subject comprising administering the compound of
7
Formula 1 or pharmaceutically acceptable salt thereof at a dose ranging from 10 to
500 mg per day. In an embodiment, the present invention provides a method of
treating synucleinopathies in a human subject comprising administering the
compound of Formula 1 or pharmaceutically acceptable salt thereof at a dose ranging
5 from 100 to 600 mg per day, preferably the dose is 200 mg to 500 mg per day and
more preferably, the dose is in the range of 300 mg to 400 mg.
In an embodiment, the present invention provides a method of treating
synucleinopathies in a human subject comprising administering the compound of
Formula 1 or pharmaceutically acceptable salt thereof at a dose selected from 10 mg,
10 50 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450 mg, 500
mg, 550 mg, 600 mg, 650 mg, 700 mg, 750 mg or 800 mg per day.
In another aspect the present invention provides a method of treating or
preventing synucleinopathies other than Parkinson’s disease in a human subject
comprising administering compound of Formula 1
N
NH
NH
Cl
CH3
H3C
O
O
Formula 1 15
or its pharmaceutically acceptable salt thereof at a dose ranging from 5 mg to
500 mg per day.
In another embodiment, the present invention provides a method of treating
or preventing synucleinopathies in a human subject comprising administering a
20 compound of Formula 1 or its pharmaceutically acceptable salt, wherein
synucleinopathy is Dementia with Lewy Bodies.
In another embodiment, the present invention provides a method of treating
or preventing synucleinopathies in a human subject comprising administering a
8
compound of Formula 1 or its pharmaceutically acceptable salt, wherein synucleinopathy is multiple system atrophy.
In another embodiment, the present invention provides a method of treating or preventing synucleinopathies in a human subject comprising administering a compound of Formula 1 or its pharmaceutically acceptable salt, wherein 5 synucleinopathy is associated with REM sleep behavior disorder.
In another embodiment, the present invention provides a method of treating or preventing synucleinopathies in a human subject comprising administering a compound of Formula 1 or its pharmaceutically acceptable salt, wherein synucleinopathy is associated with neuroautonomic dystrophies and primary 10 autonomic failure.
Suitable pharmaceutically acceptable salts of the compound of the invention may be salts of inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, and the like or of organic acids such as, for example, acetic acid, benzenesulfonic acid, methanesulfonic acid, benzoic acid, citric acid, glycolic acid, 15 lactic acid, fumaric acid, succinic acid, adipic acid, pimelic acid, suberic acid, azelaic acid, malic acid, tartartic acid, or amino acids, such as glutamic acid or aspartic acid, and the like. One or more hydrogen atoms of the compound of Formula I may be deuteriated i.e. substituted with a deuterium atom.
WIPO Publication No. WO2012098416 discloses a markush group of 20 compounds active as c-Abl kinase inhibitors and their usefulness for the treatment of cancers like chronic myelogenous leukemia (CML). Compounds of Formula I of the present invention may be prepared by the processes described in WIPO Publication No. WO2012098416.
The compound of Formula 1 can be administered orally in the form of a 25 suitable dosage form. A suitable dosage form may include tablet, pellets, capsule, sachet, pellets in sachet, pellets in capsule, powder, granules and the like. The
9
compound of Formula 1 may be formulated in oral dosage form which may include
pharmaceutically acceptable excipients which are in common knowledge of a person
skilled in the art. Remington's Pharmaceutical Sciences, Sixteenth Edition, E. W.
Martin (Mack Publishing Co., Easton, Pa., 1980) discloses pharmaceutically
5 acceptable carriers which can be used for preparation of a suitable dosage form.
WIPO Publication No. WO2017208267A1 discloses method of use of
compound of Formula I for the treatment of Parkinson’s disease. The present
invention relates to the use of the compound of Formula 1 for diseases, other than
PD, that are caused by accumulation of alpha-synuclein (aSYN) like Dementia with
10 Lewy Bodies, Pure autonomic failure, REM sleep behavior disorder, Incidental
Lewy body disease, Inherited Lewy body diseases, Lewy body dysphagia,
and multiple system atrophy.
The phrase “compound of Formula 1” is used interchangeably with the
phrase “Compound I” in the present specification and both the phrases refer to a
15 compound having a following structure:
N
NH
NH
Cl
CH3
H3C
O
O
Formula 1
The following examples serve to illustrate the invention without limiting the
invention in its scope.
Biological studies
20 Neuroprotective potential of Compound I in a rat model of synucleinopathy
Compound I was evaluated in a adeno-associated virus (AAV) AAV1/2 alpha
synuclein rat model based upon AAV1/2-mediated delivery and over-expression of
human A53T alpha-synuclein (hA53T-aSYN) in the striatonigral region of the
10
midbrain of female Sprague Dawley (SD) rats (Koprich et al, PLoS One. 7;6(3):e17698, 2011). The study was designed to assess the ability of Compound I to protect dopaminergic neurons from viral vector mediated over-expression of aSYN leading to neurodegeneration in rat model. This model is typically used as a model for aSYN-induced Parkinsonism, but may serve as a model for aSYN-induced 5 neuronal loss in general. AAV1/2 hA53T-aSYN (a viral vector delivering the capability to express human A53T mutant aSYN), or the control AAV1/2 empty vector, was injected stereotactically and unilaterally, into the striatal region of the midbrain. Rats were treated orally with Compound I (melt extrusion suspension), once daily, to provide equivalent doses of Compound I at 15, 30 and 45 mg/kg. 10
AAV1/2 hA53T-aSYN or empty AAV1/2 vector was administered sterotactically and unilaterally into the right striatal region on Day 1. Starting on Day 2 and continuing daily until Day 42, animals were fasted for 6 hr prior to oral administration of Compound I or vehicle (placebo). Food was reintroduced 60 minutes later. A total of 5 groups of animals, each group N=12, were employed. 15
Group
Initiation
Treatment
N
A
Empty Vector AAV1/2
Vehicle
12
B
AAV1/2-hA53T-aSYN
Vehicle
12
C
Compound I (15 mg/kg)
12
D
Compound I (30 mg/kg)
12
E
Compound I (45 mg/kg)
12
Animals were euthanized for post-mortem assessments on Day 43, at least 18 h after last administration of Compound I or vehicle. Brains were removed along with the terminal blood collection. As a consequence of localized synucleinopathy caused by the AAV1/2-encoded hA53T aSYN at or near the site of injection, tyrosine hydroxylase (TH)-20 expressing dopaminergic neurons in the affected striatonigral area degenerate (Koprich et al., PLoS One., 7;6(3):e17698, 2011). Number of tyrosine hydroxylase positive (TH+ve) cells within the striatonigral region of the right side of the brain
11
(injected side), were assessed using immunohistochemistry and stereological cell counting. Tyrosine hydroxylase is a critical enzyme involved in dopamine biosynthesis and thus, its presence can be used as a marker of live neurons capable of producing dopamine.
As shown in Figure 1, animals injected with AAV1/2 encoding hA53T aSYN 5 on the right side of the brain exhibited significant loss (p < 0.05) of TH+ve neurons in the striatonigral region compared to the animals that were injected, also on the right side of the brain, with the empty vector AAV1/2 incapable of expressing aSYN and also received vehicle for the treatment period. In contrast, the loss of TH+ve neurons due to synucleinopathy in animals that were injected on the right side of the 10 brain with AAV1/2 encoding hA53T aSYN and treated with different daily oral doses of Compound I was proportional to the dose of Compound I. Treatment with lower doses of Compound I (15 and 30 mg/kg) showed significant reduction (p < 0.05) in TH+ve neurons in the striatonigral region compared to the control animals that received empty AAV1/2 incapable of causing synucleinopathy. In contrast, 15 animals injected with AAV1/2 encoding hA53T aSYN and dosed with 45 mg/kg of Compound I did not show significant reduction (p > 0.05) in TH+ve neurons in the striatonigral region indicative of the neuroprotective effect of Compound I at this dose.
Optical densities of striatonigral TH-expressing neurons from the operated 20 side of the brain of animals, that received the injection of AAV1/2 encoding hA53T aSYN, were compared with that from the non-operated side of the same animals that did not show synucleinopathy. Hence for any treatment, a comparison between the operated side and non-operated side provides a clear reflection of effects on the disease state. As shown in Figure 2, in animals that received AAV1/2 (empty 25 vector), there was no significant difference in TH+ve optical densities from the left (non-operated) and right (operated) side of the brain. In contrast, animals that received AAV1/2 hA53T aSYN vector on the right side of the brain (operated side)
12
showed significantly lower (p <0.01) TH+ve optical densities compared to that from the non-operated parts of their brain. Compound I at doses of 30 and 45 mg/kg, but not at 15 mg/kg, showed significant neuroprotective effect reflected in the statistically insignificant difference (p > 0.05) between the optical densities from left (non-operated) and right (operated) sides of the brain. 5
In the example of AAV1/2 hA53T aSYN-induced synucleinopathy studied here, striatonigral TH+ve neurons capable of synthesizing dopamine were shown to be degenerated. Whether or not the neuroprotection of TH+ve dopaminergic neurons conferred by Compound I resulted in the increased production of dopamine was further examined. Hence the total dopamine levels of both the operated right side 10 and non-operated left side of the brain of the animals under study were quantified. As shown in Figure 3, dopamine levels from the operated diseased right side of the brain were always lower, as a consequence of synucleinopathy-associated neuro- degeneration, than their respective non-operated counterparts on the left side. Compound I at doses 15 and 30 mg/kg failed to restore the dopamine biosynthetic 15 capability in the operated diseased right side of the brain. In contrast, in animals dosed at 45 mg/kg of Compound I, while the dopamine levels produced from the right diseased side of the brain were still lower than that from its left counterpart, there appeared to be a gradual increase in the dopamine producing capability of the diseased part of the brain proportional to the dose of Compound I reflecting its 20 ability to not only protect neurons from degeneration but also help restore their functionality. Administration of Compound I at doses higher than 45 mg/kg may reduce this difference even further.
In conclusion, these studies collectively suggest that Compound I confers significant protection against neurodegeneration caused by synucleinopathy and help 25 restore their functionality. This neuroprotective activity of Compound I supports therapeutic applications of compound I in various disease indications attributed to synucleinopathy. ,CLAIMS:We Claim:
1. A method of treating or preventing synucleinopathies in a human
subject comprising administering a therapeutically effective amount of a compound
of Formula 1
N
NH
NH
Cl
CH3
H3C
O
O
Formula 1 5
or its pharmaceutically acceptable salt thereof.
2. A method of treating or preventing synucleinopathies in a human
subject comprising administering compound of Formula 1
N
NH
NH
Cl
CH3
H3C
O
O
Formula 1
10 or its pharmaceutically acceptable salt at a dose ranging from 0.1 mg to 1000
mg per day.
3. The method of treating or preventing synucleinopathies as in claim 1
or 2 wherein, synucleinopathy is Dementia with Lewy Bodies (DLB) or Multiple
System Atrophy (MSA) or associated with REM sleep behavior disorder.
15 4. The method of treating or preventing synucleinopathies as in claim 3
wherein, the compound of Formula 1 is administered at a dose in the range of 100
mg to 600 mg per day.
14
5. The method of treating or preventing synucleinopathies as in claim 4 wherein, the compound of Formula 1 is administered at a dose in the range of 300 mg to 500 mg per day.