Liposomal compositions for use in methods of treating Parkinson's disease

Abstract

The present invention relates to a liposome composition for use in a method for treating Parkinson's disease, the liposome composition comprising sphingomyelin in a lipid bilayer and a therapeutically effective amount of monosialotetrahexosylganglioside (GM1), wherein the therapeutically effective dose of the liposome composition is administered in a primary administration mode at least every 3 days and at most every 4 days, preferably at least every 5 days and at most every 6 days, and most preferably at least every 6 days and at most every 7 days, in the primary administration mode.

Description

[Technical Field]

[0001] The present invention relates to liposomal compositions for use in methods of treating Parkinson's disease. [Background technology]

[0002] Parkinson's disease (PD) is a brain disorder that causes unintentional or uncontrollable movements, such as tremors, rigidity, and difficulty with balance and coordination. Symptoms usually begin gradually and worsen over time. As the disease progresses, people may have difficulty walking and speaking. They may also experience mental and behavioral changes, sleep disorders, depression, memory problems, and fatigue.

[0003] The most noticeable signs and symptoms of PD occur when neurons in the basal ganglia, the area of ​​the brain that controls movement, are damaged and / or die. Normally, these neurons produce an important brain chemical known as dopamine. When neurons die or are damaged, they produce less dopamine, causing the movement problems associated with the disease.

[0004] While there is no cure for PD, medications, surgical procedures, and other therapies can often alleviate some symptoms.Medications can help treat Parkinson's symptoms by increasing levels of dopamine in the brain, by affecting other brain chemicals such as neurotransmitters that transmit information between brain cells, or by helping to control non-motor symptoms.

[0005] The main treatment for Parkinson's disease is levodopa (or L-DOPA). Nerve cells use levodopa to make dopamine to replace the brain's dwindling dopamine supplies. People usually take levodopa along with another medication called carbidopa. Carbidopa prevents or reduces some of the side effects of levodopa therapy (such as nausea, vomiting, low blood pressure, and restlessness) and reduces the amount of levodopa needed to improve symptoms.

[0006] Other medications that treat the symptoms of Parkinson's disease include dopamine agonists, which stimulate the production of dopamine in the brain; enzyme inhibitors (e.g., MAO-B inhibitors, COMT inhibitors), which increase the amount of dopamine by slowing down the enzyme that breaks down dopamine in the brain; amantadine, which helps reduce involuntary movements; and anticholinergic drugs, which reduce tremors and muscle stiffness.

[0007] Certain types of glycolipids have been identified as playing important roles in the development and treatment of PD. Glycolipids are lipids with carbohydrates attached by glycosidic bonds. Their role in living cells is to maintain cell membrane stability and facilitate cellular recognition. Glycosphingolipids are a subtype of glycolipids that contain the aminoalcohol sphingosine. They occur in the nervous system and elsewhere in the body and are involved in the metabolic and pathological changes associated with PD.

[0008] Ganglioside GM1 (monosialotetrahexosylganglioside, CAS: 37758-47-7) is a branched pentasaccharide glycosphingolipid consisting of one sialyl residue, two galactose residues, one N-acetylgalactosamine residue, and a reducing terminal glucose residue linked via a β-linkage to N-stearoylsphingosine. GM1 has important physiological properties, affecting neuronal plasticity and repair mechanisms and neurotrophin release in the brain (VASQUES, JF et al. Gangliosides in nervous system development, regeneration, and pathologies. Neural Regen. Res. 2023, Vol. 18, No. 1, pages 81 to 86). It is an amphiphilic endogenous molecule found abundantly in the outer leaflet of the membrane of all mammalian cells and is particularly abundant on neuronal plasma membranes (composing 5% ± 10% of the total lipid mass).

[0009] GM1 interacts with neighboring proteins to regulate intracellular signaling, primarily in biochemical pathways involved in neuronal differentiation, homeostasis, protection, and recovery. It has been reported to have neurotrophic properties and affect various cellular activities at the plasma membrane level and intracellularly. In the central nervous system (CNS), GM1 exhibits many ganglioside functions during development, function, and repair. It can also counteract neuroinflammation.

[0010] Higher gangliosides, such as GM1, appear only during the later stages of organogenesis and fetal development, suggesting their integral function in directing the formation of a fully functional, young CNS. The possibility of reintroducing this "young CNS" state, at least in part from a lipid-based perspective, may have therapeutic applications (XU, Y.-H. et al. Multi-system disorders of glycosphingolipid and ganglioside metabolism. J. Lipid Res. 2010, Vol. 51, No. 7, pages 1643 to 1675).

[0011] GM1 belongs to the a-series gangliosides, and its biosynthesis occurs in the Golgi apparatus. Along with more complex gangliosides, GM1 is concentrated in the pre- and postsynaptic membranes of synaptic terminals. Nuclear Ca (via high-affinity binding of gangliosides to the sodium / calcium exchanger at the nuclear envelope) is also involved. 2+ In addition to regulating homeostasis and other cellular functions, GM1 regulates neurotrophic activity by associating with receptors TrkA, TrkB, and GDNF (NEWBURN, E. et al. GM1 ganglioside enhances Ret signaling in striatum. J. Neurochem. 2014, Vol. 130, No. 4, pages 541 to 554).

[0012] The role of GM1 in both the development and potential treatment of PD has been of particular interest (CHOWDHURY, S. et al. The Key Role of GM1 Ganglioside in Parkinson's Disease. Biomolecules 2022, Vol. 12, No. 173, pages 1 to 9). GM1 deficiency disrupts the normal function of cells that depend on adequate levels of this ganglioside, leading to their gradual degeneration and eventual death. Human studies suggest that GM1 deficiency may be a contributing factor in the development of idiopathic PD. GM1 reduction has been reported in PD postmortem brains (WU, G. det al. Deficiency of ganglioside GM1 correlates with Parkinson's disease in mice and humans. J. Neurosci. Res. 2012, Vol. 90, No. 10, pages 1997 to 2008). It has also been reported that GM1 levels decrease over time in healthy human brains. However, this decrease is particularly pronounced in the brains of PD patients (HUEBECKER, M. et al. Reduced sphingolipid hydrolase activities, substrate accumulation, and ganglioside decline in Parkinson's disease. Mol. Neurodegener. 2019, Vol. 14, No. 1, pages 1 to 21).

[0013] Decreased GM1 levels have also been observed in the cerebrospinal fluid (CSF) of PD patients (compared to age-matched controls), but were not significant (17.4% reduction). A similar analysis of serum revealed a significant reduction in samples from PD patients (LEDEEN, R. et al. Systemic deficiency of GM1 ganglioside in Parkinson's disease tissues and its relationship to the disease etiology. Glycoconj. J. 2022, Vol. 39, No. 1, pages 75 to 82).

[0014] Transgenic mice lacking the GM1 family of gangliosides, either completely or partially, due to disruption of the B4galnt1 gene (GM2 synthase-B4galnt1), have been reported to develop PD features based on behavioral and neuropathological criteria. These include motor deficits (as early as 5 weeks of age) and histopathological details (such as striatal dopamine depletion, loss of dopamine neurons in the substantia nigra pars compacta, and accumulation of α-synuclein). Notably, administration of GM1 ameliorated these pathological CNS signs (WU, G. et al. Mice deficient in GM1 manifest both motor and non-motor symptoms of Parkinson's disease; successful treatment with synthetic GM1 ganglioside. Exp. Neurol. 2020, Vol. 329, 113284).

[0015] Several clinical studies have investigated the therapeutic potential of GM1 in PD cohorts. The first was a small pilot study. This was an open-label study in which 10 PD patients received 1000 mg of GM1 by intravenous infusion after the last of three baseline functional assessments, followed by self-administration of 200 mg of GM1 daily by subcutaneous injection for 18 weeks. Under these conditions, GM1 ganglioside proved safe and well-tolerated. There were no serious adverse events, and no patients developed elevated anti-GM1 antibody titers (SCHNEIDER, J.S. et al. GM1 ganglioside treatment of Parkinson's disease: an open pilot study of safety and efficacy. Neurology 1995, Vol. 45, No. 6, pages 1149 to 1154).

[0016] This study was followed up by a larger randomized, placebo-controlled study (using the same GM1 treatment regimen as above) involving 45 subjects with mild to moderate PD. This study reported significant differences between groups in Unified PD Rating Scale (UPDRS) motor scores (p = 0.0001) and activities of daily living (off-period assessment; p = 0.04) at 16 weeks (Schneider, J. et al. Parkinson's disease: improved function with GM1 ganglioside treatment in a randomized, placebo-controlled study. Neurology 1998, Vol. 50, No. 6, pages 1630 to 1636). In an open-label extension study following 5 years of GM1 use, the researchers concluded that "long-term GM1 use by PD patients is safe and may provide some clinical benefit." Participants generally had lower UPDRS motor scores (assessed during substantially defined "off" periods) (SCHNEIDER, J. Set al. GM1 ganglioside in Parkinson's disease: Results of a five-year open study. J. Neurol. Sci. 2010, Vol. 292, No. 1, pages 45 to 51).

[0017] These findings supported the initiation of a larger delayed-start study. Of 157 subjects screened, 77 were randomized to either early or delayed initiation of GM1 treatment. The GM1 treatment regimen was the same as in the previous study. A separate comparison group of 17 subjects served as a standard-of-care treatment comparator. At 24 weeks, the early-start group had a significant improvement in UPDRS motor scores, compared with a significant worsening in the delayed-start group. The early-start group also demonstrated sustained benefits compared with the delayed-start group at 72 and 120 weeks. Both groups showed significant symptom deterioration during the washout period after 2 years of GM1 treatment. This study demonstrated that long-term administration of GM1 was safe and well-tolerated, and no increase in anti-GM1 antibody titers was reported in participants (Schneider, J. Set et al. A Randomized, Controlled, Delayed Start Trial of GM1 Ganglioside in Treated Parkinson's Disease Patients. J. Neurol. Sci. 2013, Vol. 324, No. 1, pages 140 to 148). Fifteen participants from the early-start group and 14 participants from the delayed-start group (along with 11 members of the comparison group) volunteered to be part of a PET(11)C-methylphenidate imaging study. The three groups were scanned at baseline, week 24, and 1 to 2 years later. Results showed a significant delay in connectivity loss in several striatal regions in the GM1-treated group. In some cases, increased binding was detected after GM1 use (SCHNEIDER, J. et al. GM1 ganglioside in Parkinson's disease: Pilot study of effects on dopamine transporter binding. J. Neurol. Sci. 2015, Vol. 356, No. 1, pages 118 to 123).

[0018] One of the challenges in translating the therapeutic potential of GM1 from preclinical models to the clinic is the ability of this molecule to adequately reach the CNS when administered peripherally. CNS penetration is significantly limited by the amphipathic nature of GM1, which prevents it from crossing the blood-brain barrier. Therefore, novel methods for delivering GM1 to the CNS are required for clinical translation.

[0019] Furthermore, despite promising signs in research on GM1, certain side effects (adverse events) hinder the marketability of GM1 and need to be overcome. For example, because the most common mode of administration of GM1 is subcutaneous, pain, skin irritation, and severe hematomas at the injection site occur during treatment with GM1.

[0020] Due to the better bioavailability of GM1 in the CNS, the use of liposomes as drug delivery vesicles for treating PD has been proposed (WO 2019122220 A1 (INNOMEDICA HOLDING AG) 27.06.2019, pages 7 to 8). However, therapeutically effective formulations and dosage regimens are still lacking in the state of the art. Summary of the Invention [Problem to be solved by the invention]

[0021] It is therefore an object of the present invention to address these needs and provide improved treatments for PD, particularly those that reduce the adverse event profile frequently encountered with frequent subcutaneous administration. [Means for solving the problem]

[0022] The problem was solved by using a liposomal GM1 preparation having the features according to the independent claims as a medicine.

[0023] The present invention relates to a liposome composition for use in a method for treating PD, the liposome composition comprising sphingomyelin in a lipid bilayer and a therapeutically effective amount of monosialotetrahexosylganglioside (GM1), wherein the therapeutically effective dose of the liposome composition is administered in a primary administration mode at least every 4 days with at least 3 days between each administration, preferably at least every 6 days with at least 5 days between each administration, and most preferably at least every 7 days with at least 6 days between each administration, in the primary administration mode.

[0024] A liposome composition is a composition containing liposomes in addition to other components, one or more of which are encapsulated in the liposome. Liposomes are spherical vesicles with at least one lipid bilayer. Liposomes can also be multivesicular, in which a single vesicle contains one or more smaller vesicles. Liposomes have an aqueous core surrounded by a hydrophobic membrane in the form of a lipid bilayer. Liposomes limit the concentration of free components in the bloodstream and tissues, potentially providing controlled release of the encapsulated active pharmaceutical ingredient (API) over a long period of time and reducing side effects of the encapsulated component. Liposomes can also alter the tissue distribution and uptake of APIs in a therapeutically favorable manner, increasing the convenience of therapy by allowing less frequent drug administration. For example, liposomes can transport the encapsulated API directly to the site of disease. The active component can be released directly from the liposome at the treatment site. Therefore, a lower dosage of the active component is required, thereby limiting side effects.

[0025] In the case of PD, liposomes containing appropriate lipid membrane compositions enhance penetration of the blood-brain barrier, which contains blood vessels that vascularize the CNS and tightly regulate the movement of ions, molecules, and cells between the blood and the CNS. This increases the deposition of APIs encapsulated in liposomes in the CNS.

[0026] The preparation of liposome compositions containing GM1 was described in the prior art (WO2019122220A1 (INNOMEDICA HOLDING AG) 27.06.2019). In particular, the circularity of the liposomes used in the liposome composition of the present invention is 0.95 or more, in particular 0.98 to 1.00. The circularity of the liposomes in the formulation is determined by cryo-transmission electron microscopy (CryoTEM).

[0027] The lipids from the lipid membrane composition used to form the liposome of the present invention are all ubiquitous in healthy brain tissue.The liposome of the present invention contains the lipid sphingomyelin, which belongs to the group of phospholipids and sphingolipids.It accounts for about 10% of the lipids in the brain.Sphingomyelin tends to be most concentrated in the plasma membrane of living cells, especially in the outer leaflet.

[0028] As understood in the art, the lipid bilayer in liposome is a thin polar membrane composed of two layers of lipid molecules, and in aqueous medium, the polar hydrophilic ends of the lipid molecules are on the top and bottom of the membrane, and the non-polar hydrophobic ends of the lipid molecules are on the inside of the membrane.This membrane forms a sphere shell with an aqueous core.The encapsulated compound in liposome can be located in the aqueous core of liposome, or in or on the surface of the membrane.

[0029] The API of the present invention, GM1, is partially located in the membrane and partially located in the aqueous core of the liposome. As mentioned above, ganglioside GM1 is known in the art to play a major role in both the pathogenesis and potential treatment of PD. The International Union of Pure and Applied Chemistry (IUPAC) chemical name for GM1 is (2S,4S,5R,6R)-5-acetamido-2-[(2S,3R,4R,5S,6R)-5-[(2S,3R,4R,5R,6R)-3-acetamido-5-hydroxy-6-(hydroxymethyl)-4-[(2R,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxyoxan-2-yl]oxy- 2-[(2R,3S,4R,5R,6R)-4,5-dihydroxy-6-[(E,2R,3S)-3-hydroxy-2-(icosanoylamino)icos-4-enoxy]-2-(hydroxymethyl)ox-3-yl]oxy-3-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-4-hydroxy-6-[(1R,2R)-1,2,3-trihydroxypropyl]oxane-2-carboxylic acid. GM1 has the following structural formula:

[0030] [ka]

[0031] A therapeutically effective amount of monosialotetrahexosylganglioside (GM1) refers to an amount of GM1 that results in at least one of: preventing or delaying the onset of PD symptoms in a subject; improving PD symptoms in a subject; preventing or delaying the onset of PD symptoms in a subject; improving PD symptoms in a subject; or a combination thereof.

[0032] As understood in the art, a symptom is something that is felt or experienced, such as pain or dizziness. As understood in the art, a sign is an objectively observable manifestation of disease, injury, or an abnormal physiological condition that may be detected during a physical examination, examination of a patient's medical history, or diagnostic procedure. Signs and symptoms are not mutually exclusive.

[0033] These symptoms and signs include tremors, rigidity, and unintentional or uncontrollable movements, such as difficulty with balance and coordination, and difficulty walking and speaking. Symptoms are usually assessed using the Movement Disorder Society Unified PD Rating Scale (MDS-UPDRS).

[0034] The effectiveness of treatment for PD is usually measured using the MDS-UPDRS system. The system includes four parts: non-motor aspects of daily life experience are assessed in Part I of the questionnaire; motor aspects of daily life experience are assessed in Part II of the questionnaire; motor testing is performed in Part III of the clinical examination (not the questionnaire); and treatment-related motor complications are assessed in Part IV of the questionnaire. The higher the MDS-UPDRS value, the more pronounced the disease.

[0035] In the context of this invention, a dose is a measured amount of a therapeutic agent taken at one time, which may be administered over a period of time.

[0036] In the context of the present invention, a therapeutically effective dose of the liposomal composition is a dose of liposomal composition containing an amount of GM1 that results in at least one of: prevention or delay of the onset of symptoms; or amelioration of symptoms of PD in a subject.

[0037] Parkinson's disease (PD) is a brain disorder that causes unintentional or uncontrollable movements, such as tremors, rigidity, and difficulty with balance and coordination. Symptoms usually begin gradually and worsen over time. As the disease progresses, people may have difficulty walking and speaking. They may also experience mental and behavioral changes, sleep disorders, depression, memory problems, and fatigue.

[0038] According to the present invention, a dose of the liposome composition comprising GM1 is administered in the primary administration mode at least every 4 days with at least 3 days between each dose, preferably at least every 6 days with at most 5 days between each dose, and most preferably at least every 6 days with at most 7 days between each dose in the primary administration mode. For example, "at last 6 days with at most every 7 days between each dose" means that if a dose is administered on Monday, for example, the next dose in the primary administration mode will be administered the following Monday at the earliest.

[0039] The primary mode of administration can be any mode of administration deemed appropriate by one of skill in the art for administering a dose over a reasonable period of time.

[0040] It has been found that the liposomal composition as a GM1 carrier system containing sphingomyelin according to the present invention results in enhanced pharmacokinetic and therapeutic properties of encapsulated GM1.

[0041] Without wishing to be bound by theory, the liposome composition of the present invention increases the delivery of GM1 to the CNS because the liposome composition has a higher blood-brain barrier permeability than non-liposomal GM1.Furthermore, the intracellular delivery of GM1 is also improved by encapsulating it in a liposome composition.Non-liposomal GM1 remains extracellular to a greater extent because of its lower cell membrane permeability.Therefore, the biodistribution of GM1 is favorably altered by using a liposome composition to increase drug deposition in the CNS.

[0042] Furthermore, without wishing to be bound by theory, it has been found that the circulation of liposomal GM1 is prolonged compared to non-liposomal GM1 because liposomal GM1 is metabolized and excreted at a slower rate.

[0043] As a result of the increased permeability of both the blood-brain barrier and cell membranes, as well as the prolonged circulation, a lower amount of GM1 is required to achieve the desired pharmacological effect when using liposomal compositions compared to non-liposomal GM1, which in turn means that GM1 needs to be administered less frequently.

[0044] As discussed above, when GM1 is administered by injection, especially by subcutaneous injection, but also by other types of injection, strong pain, severe hematoma and skin irritation at injection site are observed.It has been found that the lower frequency of administration, which is made possible by the smaller amount of GM1 / dose and / or liposome composition, can reduce the adverse event profile in PD patients.As a result, pain, hematoma and skin irritation are less severe or not at all.

[0045] Surprisingly, the serum half-life of the liposome composition of the present invention in venous plasma was found to be longer than that typically observed for non-PEGylated (PEG: polyethylene glycol) liposome compositions containing sphingomyelin. Non-PEGylated liposome compositions containing sphingomyelin for medical use have reported serum half-lives of up to 10 hours (KRISHNA, R. et al., "Liposomal and Nonliposomal Drug Pharmacokinetics after Administration of Liposome-Encapsulated Vincristine and Their Contribution to Drug Tissue Distribution Properties," JPET. 2001, Vol. 298, pp. 1206-1212; SENIOR, J. et al., "Is the half-life of circulating liposomes determined by changes in their permeability?" FEBS Lett. 1982, Vol. 145, No. 1, pp. 109-114). According to the literature, one method for extending the half-life of liposomes in plasma is to PEGylate the liposomes (PARK, H. et al. Evolution of drug delivery systems: From 1950 to 2020 and beyond. JCR 2022, Vol. 342, pages 53 to 65). However, the liposome composition of the present invention does not require PEGylation of the liposomes, and a half-life of up to 30 hours is still observed. This indicates that the liposome composition of the present invention achieves a half-life that would not be expected by those skilled in the art compared to a non-PEGylated liposome composition containing sphingomyelin. This means that the therapeutic effect can be significantly reduced even if the administration interval is extended.

[0046] In particular, it has been found that for the majority of treated PD patients, liposomal GM1 composition needs to be administered at most every 7 days to achieve the desired therapeutic effect throughout the inter-dose period.Initial findings indicate that in certain cases, more frequent administration (e.g., every 4 days or every 6 days) is beneficial for maintaining the therapeutic effect between doses.However, this is still an improvement compared to the current dosing regimen, for example, twice a day.

[0047] The benefits, improved efficacy and reduced adverse events are demonstrated in the Examples section below.

[0048] In a preferred embodiment, the composition is administered intravenously as the primary administration method.It has been found that adverse events can be reduced not only because the amount of GM1 required is smaller and the administration frequency is lower as described above, but also when subcutaneous injection is replaced with intravenous injection.This further reduces the pain, hematoma and skin irritation of PD patients.In addition, intravenous administration can bypass the gastrointestinal barrier, for example, and therefore leads to increased bioavailability compared with other administration forms.

[0049] When the composition is administered intravenously, the dose is preferably administered over a period of 45 to 90 minutes, preferably about 1 hour. Fewer adverse events occur when the dose is administered over this period compared to shorter administration periods. In the context of the present invention, the 45 to 90 minute administration period refers to the first dose. For the second and third doses administered intravenously, a longer administration period of 90 minutes to 10 hours is recommended to avoid adverse events such as back and neck pain and potential pseudoallergic reactions (e.g., complement activation-related pseudoallergy (CARPA)).

[0050] In a preferred embodiment, administration of the primary administration mode involves administration of the secondary administration mode between doses of the primary administration mode, and the secondary administration mode is preferably oral administration. By administering an additional dose in the secondary administration mode between doses of the primary administration mode, the pharmacological effect is enhanced. For example, if the primary administration mode is weekly intravenous administration of the composition and the secondary administration mode is daily oral administration of the composition, high GM1 levels in the body can be maintained. At the same time, such an approach minimizes restrictions on quality of life. Subcutaneous injections of non-liposomal GM1 twice daily are often performed, but they result in severe side effects. In comparison, intravenous injections of liposomal GM1 once a week with an additional oral dose in between, without the need for a medical professional to be present during oral administration, are significantly more beneficial and associated with fewer side effects for patients. Surprisingly, despite the overall smaller amount of GM1 administered and the less frequent administration, this results in better pharmacological results thanks to the liposomal composition.

[0051] Preferably, the administration of the primary and / or secondary administration modes comprises a dosing regimen of equal or increasing doses of the liposomal composition containing GM1. Adverse events that may frequently occur during some of the first few doses can be offset by administering smaller amounts of the liposomal composition at earlier administrations in PD patients experiencing such side effects. As soon as side effects become less pronounced in PD patients, the dose can be increased accordingly for subsequent administrations. These side effects of the liposomal composition include back and neck pain and pseudoallergic reactions (e.g., complement activation-associated pseudoallergy (CARPA)) or humoral reactions. It should be noted that no clinically relevant adverse events have been observed with the dosing regimen according to the present invention.

[0052] As an alternative to administering a smaller amount of liposome composition to alleviate adverse events occurring during intravenous administration, the infusion flow rate can be adjusted so that less liposome composition enters the body per unit time. Furthermore, both the dosage and administration rate of the liposome composition can be adjusted during the same administration. The choice of dosage and administration rate depends heavily on patient feedback. For example, if a patient complains of pain in the neck, back, or around the injection site during administration, the administered amount and / or administration rate can be adjusted accordingly. Furthermore, the infusion can be suspended until the symptoms of the adverse event subside.

[0053] In the context of the present invention, dosing regimen is the schedule of the dosage of therapeutic agent per unit time, including dosing interval, amount of therapeutic agent per administration, and administration period, for example, in the case of intravenous injection.To treat patients, for example, when dosing interval varies, several dosing regimens of the same dosing mode can be used consecutively.In addition, for different dosing modes, several dosage regimens can also be used in parallel.

[0054] Preferably, the primary administration mode includes a second dose of the liposomal composition containing a lower amount of GM1 than the first dose, and increasing third and subsequent doses.

[0055] In a preferred embodiment, the composition is administered periodically every 7 days with 6 days between each administration in the primary administration regimen.

[0056] In another preferred embodiment, the therapeutically effective dose of GM1 in the liposomal composition is 300 mg to 800 mg, preferably 600 mg to 750 mg, and most preferably about 720 mg, which has been found to provide optimal pharmacological efficacy without causing serious side effects.

[0057] In a more preferred embodiment, the liposome composition further comprises cholesterol, preferably sphingomyelin and cholesterol, in a 1:1 molar ratio. Liposomes containing sphingomyelin and cholesterol exhibit enhanced circulation life and CNS bioavailability. They have improved pharmacokinetics and therapeutic properties. They are biocompatible and biodegradable. In certain cases, elevated cholesterol levels may be observed after administration of liposomal cholesterol-containing compositions, but these elevated cholesterol levels return to normal levels without any resulting adverse events.

[0058] In a preferred embodiment, the therapeutically effective amount of GM1 in the liposomal composition in a single-dose primary administration regimen is selected to produce a venous plasma GM1 concentration of 50 μg / ml to 1200 μg / ml, particularly 75 μg / ml to 600 μg / ml, and more particularly 100 μg / ml to 400 μg / ml, reached within 1 to 7 hours, preferably 3 to 5 hours, and most preferably within 4 hours after the start of administration. This range of venous plasma GM1 concentration has been found to produce optimal pharmacological effects without causing serious side effects. At these values ​​of venous plasma GM1 concentration, GM1 concentrations were found to be significantly higher after 96 hours compared to baseline (see the Examples section below). This supports the promise of a long-circulating drug.

[0059] In yet another preferred embodiment, the liposomes of the liposome composition have an average diameter of 10 nm to 70 nm, preferably 30 nm to 70 nm, more preferably 40 nm to 65 nm, as measured by dynamic light scattering; and / or an average diameter of 10 nm to 50 nm, preferably 20 nm to 50 nm, more preferably 30 nm to 40 nm, as measured by CryoTEM.

[0060] "Measured by dynamic light scattering" (DLS) means that DLS was performed on samples with lipid concentrations of 20 mg / ml to 30 mg / ml, which were diluted 1 / 19 with phosphate-buffered saline (PBS) or milliQ HO to an instrument extinction coefficient of approximately 6. DLS was measured on a Malvern Zetasizer Nano device at 25 °C and a scattering angle of 0°. Instrument control and data analysis were performed using Malvern's Zetasizer software (version 7.11). Particle size (hydrodynamic diameter) was determined using the Stokes-Einstein equation.

[0061]

number

[0062] where k is the Boltzmann constant, T is the absolute temperature, η is the dispersant viscosity, and D is the diffusion coefficient. The viscosity was determined using Zetasizer software and was 0.8872 cP. The dispersant refractive index was 1.330. D was obtained by fitting the autocorrelation function with an appropriate algorithm. Cumulant analysis is a simple method for analyzing the autocorrelation function generated by DLS experiments to produce the mean particle size (Z-ave) and polydispersity index (PDI). The calculation is defined in ISO 13321 (1996) and ISO 22412 (2008). The primary result from a DLS experiment is the particle size intensity distribution. The intensity distribution is naturally weighted according to the scattering intensity. The size distribution is displayed as a plot of the relative intensity of light scattered by the particles (on the Y-axis) against various logarithmically spaced size classes (on the X-axis). A transparent disposable Zeta cell with a 10 mm path length was used for the measurements. Typically, but not necessarily, liposomes in the compositions of the present invention will fall within the numerical range of sizes measured by the described methods.

[0063] "Measured by CryoTEM" means that the sample was subjected to cryo-transmission electron microscopy (CryoTEM). Liposome samples were appropriately diluted, vitrified, and prepared on a grid (Formvar and Carbon) at an accelerating voltage of 200 kV. Images were acquired at 20,000x, 40,000x, and 80,000x magnifications using a JEOL JEM-2100F TVIPS TemCam F415MP CryoTEM camera. Particle identification and sizing were performed by semi-automated image processing using Vironova Analyzer Software (Vironova, Sweden). Briefly, a series of random images at the same magnification were imported. Only liposome particles that were completely located within the image boundaries and had a distinct membrane were detected. Identified objects were analyzed for sphericity, circularity, and monolayerness. All images were batch-processed with identical thresholds and settings, and more than five to 18 images were accumulated for each sample, corresponding to a number of analyzed particles ranging from 6 to 1560–1178. The average value has a standard deviation of about 10 nm.

[0064] Typically, but not necessarily, liposomes in the formulations of the present invention fall within a range of sizes measured by both methods, with diameters measured by CryoTEM generally being smaller than those measured by DLS.

[0065] In the context of the present invention, the term "and / or" in relation to measuring liposome diameter should be understood as an inclusive or, meaning that all liposome compositions are included, where the liposomes typically, but not necessarily, meet either or both conditions.

[0066] The liposomes of the present invention and the indicated sizes of the compositions have been successfully shown to be more stable than those known in the art. Such small diameter liposomes are less rapidly and to a lesser extent opsonized than their larger counterparts and are not as rapidly cleared by the reticuloendothelial system. Larger liposomes are also more likely to fuse or interact with other liposomes or particles.

[0067] As a result, the serum half-life of the liposome composition of the present invention in humans is significantly longer than that of liposome compositions with higher liposome diameters known in the art. Furthermore, the smaller liposome diameter allows for accelerated passage through the blood-brain barrier. Thus, the composition provides higher drug exposure in the CNS at a given dose (HERSH, A.M. Crossing the Blood-Brain Barrier: Advances in Nanoparticle Technology for Drug Delivery in Neuro-Oncology. Int. J. Mol. Sci. 2022, Vol. 23, pages 1 to 28).

[0068] In a preferred embodiment, the liposome composition comprises phosphate buffered saline (PBS) at a pH of about 6.8, which corresponds to a physiologically tolerable pH.

[0069] In another preferred embodiment, the liposome composition comprises at least one of a pharmaceutically acceptable additive, carrier, excipient, and diluent.

[0070] In the context of the present invention, an excipient is a substance added to a composition along with an API to impart a particular quality to the composition. An excipient has little or no therapeutic value but is necessary for the manufacture of a particular dosage form. An excipient may serve any one or any combination of the following purposes: to provide bulk to the composition; to facilitate drug absorption or solubility and other pharmacokinetic considerations; to assist in the handling of the API during manufacturing; to provide stability and prevent degradation.

[0071] In the context of the present invention, a carrier (also known as a drug carrier or drug vehicle) is a substance used in the drug delivery process that serves to improve the selectivity, efficacy, and / or safety of drug administration. Common types of carriers include liposomes, micelles, microspheres, and nanoparticles.

[0072] In the context of this invention, excipients are substances that are formulated with an API and are included for the purposes of long-term stabilization, bulking up a solid formulation containing a small amount of potent active ingredient, or to impart therapeutic enhancements to the API such as facilitating drug absorption, reducing viscosity or enhancing solubility or enhancing bioavailability.

[0073] In the context of this invention, a diluent (also called a filler, dilutant, or thinner) is an ingredient in a pharmaceutical formulation that lacks pharmacological activity but is pharmaceutically necessary or desirable. It is particularly useful for increasing the bulk of a potent drug substance at a mass that is too small for a dosage to permit manufacture or administration.

[0074] In a preferred embodiment, the method for treating PD is selected from the following: reducing tremor in PD patients, increasing physical activity, increasing walking speed, improving walking ability, improving movement control, and improving speech changes.The above complaints are common symptoms in PD patients.It has been found that treating the affected area of ​​the CNS with the liposome composition of the present invention can significantly reduce these symptoms when the severity of the symptoms is assessed using the MDS-UPDRS system.

[0075] Another aspect of the present invention relates to a liposome composition for use in a method for treating Parkinson's disease (PD), the liposome composition comprising sphingomyelin in a lipid bilayer and a therapeutically effective amount of monosialotetrahexosylganglioside (GM1), wherein the treatment is at least partial reversal of PD, and optionally, the treatment comprises a dosing regimen in which a therapeutically effective dose of the liposome composition is administered in a primary administration mode up to every four days with at least three days between each dose, preferably up to every six days with at least five days between each dose, and most preferably up to every seven days with at least six days between each dose.

[0076] In the context of the present invention, at least partial reversal of PD is at least a partial decrease in at least one symptom or sign of the disease, wherein at least one symptom or sign does not increase substantially compared to baseline.

[0077] Surprisingly, it has been found that treatment of PD patients with the liposomal compositions of the present invention for 24 weeks not only halts the negative progression of PD, but can significantly reduce symptoms and signs, even when the liposomal compositions of the present invention are administered at 7-day intervals.

[0078] The present invention is further illustrated by the following examples, which are not intended to limit the scope of the invention in any way.

[0079] The preparation of liposome compositions containing GM1 has been described in the prior art (WO2019122220A1 (INNOMEDICA HOLDING AG) 27.06.2019). In particular, the circularity of the liposomes used in the liposome composition of the present invention is 0.95 or more, in particular 0.98 to 1.00.

[0080] The liposome composition containing GM1 used for intravenous administration in the following examples may be referred to hereinafter as "TLN" or "talinuren." [Brief explanation of the drawings]

[0081] [Figure 1a] Figure 1a-1c: Striatal levels of dopamine and its metabolites in mice in the MPTP-PD model: levels of dopamine (DA, Figure 1a), 3,2-dihydroxyphenylacetic acid (DOPAC, Figure 1b) and homovanillic acid (HVA, Figure 1c) determined by HPLC, n=5–10, bars indicate mean and standard deviation, one-way ANOVA treatment group p-values ​​*<0.05, **<0.01; [Figure 1b] Figure 1a-1c: Striatal levels of dopamine and its metabolites in mice in the MPTP-PD model: levels of dopamine (DA, Figure 1a), 3,2-dihydroxyphenylacetic acid (DOPAC, Figure 1b) and homovanillic acid (HVA, Figure 1c) determined by HPLC, n=5–10, bars indicate mean and standard deviation, one-way ANOVA treatment group p-values ​​*<0.05, **<0.01; [Figure 1c] Figure 1a-1c: Striatal levels of dopamine and its metabolites in mice in the MPTP-PD model: levels of dopamine (DA, Figure 1a), 3,2-dihydroxyphenylacetic acid (DOPAC, Figure 1b) and homovanillic acid (HVA, Figure 1c) determined by HPLC, n=5–10, bars indicate mean and standard deviation, one-way ANOVA treatment group p-values ​​*<0.05, **<0.01; [Figure 2] Reverse rotarod test performance in C9orf72 mice: Reverse rotarod test starting treatment from P110 to P151 (15 mg / kg saline or talinuren (TLN), intravenous administration, every other day, dotted box), n=5–7 / group. Values ​​represent the mean and standard error of the mean. Differences were assessed using two-way ANOVA with Bonferroni's multiple comparison test, ****p<0.0001; [Figure 3] Hematoxylin and eosin (H&E) and NADH staining of gastrocnemius muscle from a C9orf72 mouse at P170. Left: H&E staining of gastrocnemius muscle. Black arrows indicate keratinized fibers. White arrows indicate muscle fibers with central nuclei, indicating denervation. Right: NADH staining showing type I (dark) and type II (light) muscle fibers. n=3 per group. Samples were stained and imaged using the same settings. [Figure 4] Reverse rotarod test performance of SOD1-G93A mice treated starting at P110–P121 and P160–P170 (15 mg / kg saline or talinuren ganglioside (TLN), intravenous, every other day, dotted box), n=4–6 / group. Values ​​represent the mean and standard error of the mean. Differences were assessed using two-way ANOVA with Bonferroni's multiple comparison test, ****p<0.0001; [Figure 5a] Figure 5a+5b Difference in MDS-UPDRS motor score (Fig. 5a) and total score (Fig. 5b) for each patient (P1-P7; P9-P12) between baseline and 8-week post-treatment assessment. 5a: Change in UPDRS motor score ('off' state after 12 hours of discontinuing standard of care medication) (R); 5b: Change in UPDRS total score (S); [Figure 5b] Figure 5a+5b Difference in MDS-UPDRS motor score (Fig. 5a) and total score (Fig. 5b) for each patient (P1-P7; P9-P12) between baseline and 8-week post-treatment assessment. 5a: Change in UPDRS motor score ('off' state after 12 hours of discontinuing standard of care medication) (R); 5b: Change in UPDRS total score (S); [Figure 6a] Figure 6a+6b: Difference in MDS-UPDRS motor score (Figure 6a) and total score (Figure 6b) for each patient (P1-P7; P9-P12) between baseline and 24-week assessment. 6a: Change in UPDRS motor score ('off' state after 12 hours of discontinuing standard of care medication) (R); 6b: Change in UPDRS total score (S); [Figure 6b] Figure 6a+6b: Difference in MDS-UPDRS motor score (Figure 6a) and total score (Figure 6b) for each patient (P1-P7; P9-P12) between baseline and 24-week assessment. 6a: Change in UPDRS motor score ('off' state after 12 hours of discontinuing standard of care medication) (R); 6b: Change in UPDRS total score (S); [Figure 7] Plasma GM1 concentrations (U) over time (T) and mean values ​​at each measurement time point for all nine patients in the dose-intensified group; [Figure 8a]Figure 8a+8b: TLN-DiI uptake in cells of mouse brain: 8a: Substantia nigra after 48 hours (TLN-DiI, iv, 30 mg / kg). DAPI (nuclear DNA, blue), DiI (dye, red). 8b: Motor cortex after 24 hours (TLN-DiI, iv, 15 mg / kg). Scale bar = 50 μm. White arrows indicate DiI-positive cells. [Figure 8b] Figure 8a+8b: TLN-DiI uptake in cells of mouse brain: 8a: Substantia nigra after 48 hours (TLN-DiI, iv, 30 mg / kg). DAPI (nuclear DNA, blue), DiI (dye, red). 8b: Motor cortex after 24 hours (TLN-DiI, iv, 15 mg / kg). Scale bar = 50 μm. White arrows indicate DiI-positive cells. [Figure 9] GM1 levels in rat brain (V) determined by LC-MS / MS after a single daily intravenous dose of 12.3 mg / kg body weight of GM1 (free or talinurene form). (t-test with Welch's correction, p-value < 0.05), bars indicate mean and standard deviation, n = 5; [Figure 10] Changes in the Parkinson's Disease Questionnaire (PDQ-39) are the mean scores (X) of the eight daily living domains over time at baseline (time t = 0 week (w)), after 8 weeks and 24 weeks of treatment (the mean, median, maximum and minimum values ​​are shown from patients P1 to P12); W: time in weeks (w). DETAILED DESCRIPTION OF THE INVENTION

[0082] Talinurene has been studied in several preclinical models of neurological disorders. Efficacy studies have been conducted in animal models of different movement disorders, such as the MPTP mouse model of PD and the C9orf72 and Sod1 mouse models of amyotrophic lateral sclerosis. [Example]

[0083] Example 1: Talinurene in a PD model MPTP (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine) is a neurotoxin +It is a prodrug for PD-1, which causes the permanent symptoms of PD by destroying dopaminergic neurons in the substantia nigra of the brain.

[0084] Studies in the MPTP mouse model of PD, in which dopaminergic neurodegeneration is induced by a neurotoxin, have yielded mixed results. MPTP was administered intraperitoneally (ip) at a dose of 20 mg / kg in saline twice daily, 4 hours apart, for two consecutive days (days 1 and 2), for a total dose of 80 mg / kg. The MPTP dose was 10 mL / kg, resulting in a concentration of 1.6 mg / mL of pure MPTP active compound (after a salt correction factor was reduced). MPTP administration began in the morning (8–9 AM).

[0085] Mice were administered talinurene per os (po) daily for 14 days (days 1-14). The dose was 5 ml / kg. On the MPTP administration day (days 1-2), compound administration was performed in the evening (6-8 PM). On day 3, compound administration was performed during the day (1-2 PM). On days 4-14, compound administration was performed starting at 7 AM.

[0086] Figure 1 shows striatal levels of dopamine (DA, Figure 1a), 3,2-dihydroxyphenylacetic acid (DOPAC, Figure 1b), and homovanillic acid (HVA, Figure 1c) measured by high-performance liquid chromatography (HPLC). Five to ten subjects were studied for each treatment group, and the bars represent the mean and standard deviation in wet tissue. Groups received the following treatments: A:Sham; B: MPTP + PBS; C: MPTP+BV (base vesicle) 30 mg / kg, po; D: MPTP+GM1 30 mg / kg, i.p.; E: MPTP+GM1 30 mg / kg, po; F:MPTP+TLN 7.5 mg / kg, po; G:MPTP+TLN 15 mg / kg, po; and H:MPTP+TLN 30mg / kg, p.o. In initial studies using oral doses of 7.5 mg / kg (F) and 15 mg / kg (G), talinurene showed partial rescue of dopamine and dopamine metabolite levels in the substantia nigra.

[0087] Example 2: Talinuren in ALS models Amyotrophic lateral sclerosis (ALS) is a progressive neurological disease that affects nerve cells in the brain and spinal cord, causing loss of muscle control.

[0088] To evaluate potential therapeutic effects in ALS, we used two preclinical models harboring the most common mutations in familial ALS. In the C9orf72 and SOD1 mouse models of amyotrophic lateral sclerosis, talinuren ganglioside improved the mice's motor performance and reduced ER stress at the cellular level. Liposome uptake was demonstrated in in vitro cultures of cortical neurons and in vivo in spinal cord motor neurons. More importantly, talinuren ganglioside treatment in the C9orf72 transgenic model of ALS demonstrated significant effects on motor performance. Motor function and coordination were significantly improved not only when intravenous talinuren treatment was initiated early in the disease, but also when it was initiated at more advanced stages.

[0089] Figure 2 shows the reverse rotarod test performance of C9orf72 mice. This test is used to assess motor coordination and balance in rodents. The apparatus consists of a motorized circular rod that rotates at a constant or increasing speed. Rodents naturally try to stay on the rotarod to avoid falling onto the platform. The latency to fall (L) is measured in seconds. Figure 2 shows the following curves:

[0090] I: wild-type mice injected with saline only; J: wild-type mouse injected with talinuren; K: C9orf72 mice injected with saline alone; M: C9orf72 mice injected with talinuren.

[0091] Treatment began on postnatal day 110 (P110) and continued until P151 (dotted box in Figure 2). When intravenous treatment was initiated and continued early in the disease, a prolonged latency to fall (L) was observed in C9orf72 mice injected with talinuren (M), near wild-type (WT) levels. The elevated L was maintained until the end of treatment at P151. This effect remained highly significant throughout the treatment period. Muscle strength was also improved in C9orf72 mice treated with talinuren ganglioside compared with the C9orf72 saline group in a suspension wire test performed on postnatal day 140 (P140).

[0092] These changes in the mice's physical condition were supported by microscopic observation of spinal cord sections (Figure 3). Talinuren ganglioside treatment reduced metabolic stress, as indicated by downregulation of a marker for endoplasmic reticulum stress (BiP, glucose-regulated protein), known to be involved in ALS progression.

[0093] Neuronal inclusions of PolyGA, a protein translated from repeat expansions in C9orf72 mutations, are abundant in ALS patients and have been detected in postmortem tissue. PolyGA aggregation correlates with the unfolded protein response and causes behavioral deficits through inflammation and protein sequestration, likely contributing to prodromal symptoms and disease progression in C9orf72 patients (Schludi, MH et al. Spinal poly-GA inclusions in a C9orf72 mouse model trigger motor deficits and inflammation without neuron loss. Acta Neuropathol. 2017, Vol. 134, No. 2, pages 241 to 254). There is a significant reduction in large PolyGA aggregates in talinurene-treated animals. In skeletal muscle, denervation of the neuromuscular junction in mice carrying the C9orf72 mutation has been previously described (LIU, Y. et al. C9orf72 BAC Mouse Model with Motor Deficits and Neurodegenerative Features of ALS / FTD. Neuron 2016, Vol. 90, No. 3, pages 521 to 534). In preclinical studies, H&E staining of gastrocnemius muscle at P170 revealed clusters of keratinized fibers and myofibers with central nuclei, indicating myofiber degeneration and denervation, in C9orf72-untreated mice but not in WT mice or C9orf72 mice treated with talinuren (see Figure 3, left panel). Furthermore, nicotinamide adenine dinucleotide (NADH) staining demonstrated type I and type II myofibers in WT mice. Myofibrillar structure was reduced in untreated C9orf72 mice, indicating mitochondrial or sarcoplasmic reticulum abnormalities and neurogenic processes (see right panel of Figure 3). C9orf72 animals treated with talinuren displayed near-WT characteristics.

[0094] The therapeutic effect of intravenously administered talinuren was also observed in a preclinical ALS model harboring an SOD1 mutation. Figure 4 shows the following curves:

[0095] N: wild-type mice injected with saline only; O: wild-type mice injected with talinuren; P: C9orf72 mice injected with saline only; Q: C9orf72 mice injected with talinuren.

[0096] The two treatment intervals, between P110 and P121 and between P161 and P171, are marked by dotted boxes in Figure 4. Treatment for 10 days, starting on postnatal day P110, resulted in improved rotarod performance in animals carrying the mutation. Although the effect diminished over time, treated animals maintained their improved performance. However, a second treatment interval at P160 failed to enhance performance.

[0097] Example 3: Clinical trial results A phase I clinical trial is being conducted as an open-label, single-ascending dose study (n = 3) followed by multiple doses at the maximum adequate dose (n = 9) (Neurologisches Institut Konolfingen; ClinicalTrials.gov identifier: NCT04976127). The primary objective was to demonstrate the safety of intravenous TLN administration in patients with Parkinson's disease. Secondary objectives were to determine the maximum adequate dose based on the safety profile and preliminary efficacy, and to determine the pharmacokinetic profile.

[0098] Dose escalation The administered dose in all three PD patients was increased weekly to the maximum intended dose of 720 mg without any serious adverse events, dose delays, or dose modifications. Therefore, the maximum preferred dose for the dose intensification part of the study was determined at the highest dose level (720 mg), confirming the expected good tolerability of the drug.

[0099] dose fortification Nine additional patients were enrolled in the trial and treated for 8 weeks at this established therapeutic dose. All nine patients completed the 8-week course of talinurene treatment. Pharmacokinetic blood samples measuring serum GM1 were collected at 10 different time points during the first and subsequent days of talinurene administration. All samples were analyzed, and the results are presented below.

[0100] Voluntary extension of treatment All patients already enrolled in the trial expressed a desire to continue weekly talinuren treatment, as they subjectively experienced benefit from the treatment. Because talinuren was generally well tolerated, the trial was extended twice (Extension 1: 8 weeks; Extension 2: 16 weeks). This allowed for continued benefit from talinuren for patients while collecting additional safety data on the 720 mg repeated dose of the study drug.

[0101] Safety / Tolerability Treatment was generally well tolerated, with no serious adverse events. Some patients experienced CARPA (complement activation-associated pseudoallergy), which was not considered a serious adverse event and conformed to the clinical trial protocol's safety definition (patients experienced neck pain and headache and were rapidly stopped with a reduction in infusion rate). CARPA is a known phenomenon in nanotechnology investigational drugs, and all CARPA experienced in this study were fully reversible.

[0102] Efficacy after 8 weeks of treatment The efficacy analysis after 8 weeks of treatment contains data from all patients (11 of 12 patients) treated with weekly infusions of 720 mg of the active pharmaceutical ingredient GM1 ganglioside in a liposomal formulation of the present invention over the entire study period of 8 weeks of treatment. One patient (patient number 8) received only 2 weeks of treatment and dropped out of the study.

[0103] To measure efficacy, two assessments were considered: 1) MDS-UPDRS motor score (Part III). According to the literature (SCHNEIDER, J. Set et al. A Randomized, Controlled, Delayed Start Trial of GM1 Ganglioside in Treated Parkinson's Disease Patients. J. Neurol. Sci. 2013, Vol. 324, No. 1, pages 140 to 148), the assessment was performed in an "off" medication state, meaning that standard treatment was postponed for 12 hours. Assessments were performed at two time points: the first was a baseline (BL) measurement before the start of treatment, and the second was one week after the final treatment during the final assessment (FA) 8 weeks later.

[0104] 2) MDS-UPDRS total score. Part III was measured as described in 1). Parts I, II, and IV correspond to the standard assessment procedure of the MDS-UPDRS using a questionnaire. Assessments were performed during the first talinuren treatment (BL) and one week after the final treatment (FA).

[0105] Table 1 shows the data points for each patient according to the clinical trial database separately for both efficacy measures considered (motor score in the "off" state and MDS-UPDRS total score). Columns 3 and 6 show the difference between the baseline (BL) assessment and the final assessment (FA) after 8 weeks for each patient. The motor score and total score were reduced by an average of 6.7 and 12.2 points, respectively. Using the nonparametric Wilcoxon rank-sign test for paired samples, the difference is statistically significant at the 1% level. Clinically, this difference is also considered relevant, since the "minimally clinically important difference" (MCID) according to the literature is an improvement of approximately 4.3 to 5 points.

[0106] [Table 1]

[0107] Figure 5a and Figure 5b show the difference between baseline (BL) and the final assessment (FA) after 8 weeks for the MDS-UPDRS motor score (Part III) and MDS-UPDRS total score for each patient (P1–P7; P9–P12), respectively, in descending order. The labels on the y-axis mean the following:

[0108] R: Change in UPDRS motor score (12 hours "off" standard drug treatment); S: Change in UPDRS total score.

[0109] Efficacy after 24 weeks of treatment Due to promising results after 8 weeks of treatment, treatment was continued for a total treatment period of 24 weeks. The efficacy analysis after 24 weeks of treatment contains data from patients P1-P7 and P9-P12, who received weekly infusions of 720 mg of the active pharmaceutical ingredient GM1 ganglioside in a liposomal formulation of the present invention.

[0110] [Table 2]

[0111] Table 2 shows the data points for patients P1-P7 and P9-P12 for both efficacy measures considered (motor score in the "off" state and MDS-UPDRS total score). Columns 3 and 6 show the difference between the baseline (BL) assessment and the final assessment (FA) after 24 weeks for each patient. Motor scores and total scores were reduced by an average of 7.5 points and 12.2 points, respectively. These results indicate that there was a further improvement in MDS-UPDRS motor scores (Part III) after 24 weeks of total treatment when compared to the results after 8 weeks of total treatment.

[0112] Figures 6a and 6b show the difference between baseline (BL) and the final assessment (FA) after 24 weeks for the MDS-UPDRS motor score (Part III) and MDS-UPDRS total score for each patient (P1–P7; P9–P12), respectively, in descending order. The labels on the y-axis indicate the following:

[0113] R: Change in UPDRS motor score (12 hours "off" standard drug treatment); S: Change in UPDRS total score.

[0114] Efficacy after 56 weeks of treatment As with the efficacy analysis after 24 weeks of treatment, treatment continued for a total of 56 weeks for the same patients, except for patient P11, who again received weekly infusions of 720 mg of the active pharmaceutical ingredient GM1 ganglioside in a liposomal formulation of the present invention.

[0115] [Table 3]

[0116] The data in Table 3 show that values ​​remained at low levels compared to baseline values ​​after 56 weeks.

[0117] Parkinson's Disease Questionnaire (PDQ-39) In addition to the value used to determine Parkinson's disease severity, as determined using the Unified Parkinson's Disease Rating Scale (UPDRS), treatment efficacy was also analyzed using the Parkinson's Disease Questionnaire (PDQ-39). This questionnaire, which contains 39 questions, assesses the frequency with which people with Parkinson's disease experience difficulties across eight domains (dimensions) of daily life: mobility, activities of daily living (ADL), emotional well-being, stigma, social support, cognition, communication, and physical discomfort. The total score for each domain ranges from 0 (patient never has difficulty) to 100 (patient always has difficulty).

[0118] Questionnaires were administered to the 12 patients (P1-P12) at baseline (t=0), after 8 weeks of treatment, and after 24 weeks of treatment (patient P8 did not complete the questionnaire after 24 weeks). At each time point (t=0, 8, and 24 weeks), mean values ​​from the eight domains of daily living were calculated for each patient. From the values ​​obtained for the 12 patients, mean, median, maximum, and minimum values ​​were calculated for each time point.

[0119] Figure 10 shows these values ​​(X) over time (W; weeks (w)). The mean values ​​(circles) show the reduction in PDQ-39 scores from 20.1 at time 0 to a score of 17.2 after 8 weeks and to a score of 15.6 after 24 weeks. The median values ​​(triangles), maximum values ​​(squares) and minimum values ​​(diamonds) show the same trend over time. A particularly strong improvement was observed in the patient with the most severe score initially (Patient 6: 33.9). The maximum values ​​both after 8 weeks (28.7) and after 24 weeks (23.3) belong to this patient.

[0120] These results demonstrate that significant improvements are observed not only in the MDS-UPDRS assessment procedure, but also in a second assessment procedure (PDQ-39 scoring system) in patients treated with the liposomal compositions and dose regimens according to the present invention.

[0121] Pharmacokinetics / Pharmacodynamics Samples from nine patients receiving weekly infusions of 720 mg talinuren (dose intensification part) were collected during 10 time points during the first infusion (0 min, 5 min, 30 min, 1 h, 4 h, 8 h, 24 h, 48 h, 72 h, 96 h).

[0122] [Table 4]

[0123] Table 4 shows the time to reach peak plasma concentration (t max ), maximum plasma concentration (c max ) and the half-life of GM1 in plasma (t 1 / 2) are shown. Also shown are the mean, standard deviation (SD), minimum, median, maximum, coefficient of variation (CV%), geometric mean, and geometric mean of the coefficient of variation for these values. Figure 7 shows the GM1 concentration over time in the plasma of all nine patients and the mean values ​​for each measurement time point. The labels on the axes mean the following:

[0124] T: time after initiation of infusion (h); U: GM1 concentration (ng / ml).

[0125] GM1 was measured in all samples. Most patients reached peak intravenous plasma concentrations after 4 hours. The infusion lasted 90 minutes, and no sampling was performed at that time. Therefore, peak plasma concentrations may also be reached earlier. Compared to baseline, significantly higher levels of GM1 were still present after 96 hours, confirming the prediction of a long-circulating drug.

[0126] The mean half-life was slightly lower than expected, but there were some individual high values ​​(patient 11) with a large standard deviation. With the currently measured mean half-life and a frequency of only one dose per week, there may not be any accumulation in plasma. Therefore, it may be interesting to test different dosing frequencies to reach steady-state conditions. The mean volume of distribution was relatively low, which may suggest that GM1 is primarily located in the blood. This may also mean that GM1 is widely distributed in the central nervous system and not in other hydrophobic parts of the body.

[0127] Example 4: Brain penetrance Talinurene delivery to the brain was evaluated in mice. Fluorescently labeled talinurene (TLN-DiI) was administered to mice, and dye accumulation was measured 24 or 48 hours later.

[0128] Figure 8a shows TLN-DiI uptake in cells of the substantia nigra of a mouse brain 48 hours after administration (TLN-DiI, iv, 30 mg / kg). The left image of Figure 8a shows both nuclear DNA (DAPI, blue) and DiI dye (red), while the right image of Figure 8a shows only DiI dye (red). Figure 8b shows TLN-DiI uptake in cells of the motor cortex of a brain 24 hours after administration (TLN-DiI, iv, 15 mg / kg). DiI signals were detected in cells of the substantia nigra as well as the motor cortex (white arrows).

[0129] A study using CD-1 rats investigated the accumulation of talinuren's active pharmaceutical ingredient (GM1) in the rat brain. Each animal received a daily intravenous dose of either free (GM1) or liposomal GM1 (TLN) at 12.3 mg / kg body weight for four consecutive days. Brain GM1 levels were quantified 24 hours after the final dose by liquid chromatography-mass spectrometry / mass spectrometry (LC-MS / MS).

[0130] Figure 9 shows GM1 levels in rat brains (V) after 4 days of treatment. The levels were significantly increased in the talinurene-treated group (liposomal GM1; TLN) compared with the free GM1-treated group. This indicates that GM1 is delivered to the brain more efficiently when it is incorporated into liposomes.

Claims

1. 1. A liposome composition for use in a method for treating Parkinson's disease (PD), the liposome composition comprising sphingomyelin in a lipid bilayer and a therapeutically effective amount of monosialotetrahexosylganglioside (GM1), wherein a therapeutically effective dose of the liposome composition is administered in a primary administration mode with at least three days between each administration and at most every four days, preferably in a primary administration mode with at least five days between each administration and at most every six days, and most preferably in a primary administration mode with at least six days between each administration and at most every seven days.

2. The composition of claim 1 , wherein the composition is administered intravenously in the primary mode of administration.

3. 3. The composition of claim 2, wherein the dose is administered over a period of 45 to 90 minutes, preferably about 1 hour.

4. 10. The composition of any one of the preceding claims, wherein said administration in said primary mode of administration is accompanied by administration in a secondary mode of administration between doses of said primary mode of administration, said secondary mode of administration being preferably oral administration.

5. 10. The composition of any one of the preceding claims, wherein the administration of the primary and / or secondary modes of administration comprises a dosing regimen of equal or increasing doses of the liposomal composition comprising GM1.

6. The composition of any one of claims 1 to 4, wherein the primary administration mode comprises a second dose of the liposome composition containing a lower amount of GM1 than the first dose, and / or the second dose is administered at a lower flow rate than the first dose.

7. 10. The composition of any one of the preceding claims, wherein the composition is administered periodically every 7 days with 6 days between each administration in the primary administration regimen.

8. 10. The composition of any one of the preceding claims, wherein the therapeutically effective dose of GM1 in the liposomal composition is between 300 mg and 800 mg, preferably between 600 mg and 750 mg, most preferably about 720 mg.

9. 10. The composition of any one of the preceding claims, wherein the liposome composition further comprises cholesterol, preferably sphingomyelin and cholesterol in a 1:1 molar ratio.

10. 10. The composition according to any one of the preceding claims, wherein the therapeutically effective amount of GM1 in the liposome composition in the primary administration mode of a single dose is selected to result in a venous plasma GM1 concentration of 50 μg / ml to 1200 μg / ml, particularly 75 μg / ml to 600 μg / ml, more particularly 100 μg / ml to 400 μg / ml, reached within 1 to 7 hours, particularly within 3 to 5 hours, more particularly within 4 hours after the start of administration.

11. The liposomes of the liposome composition are - have an average diameter, as measured by dynamic light scattering, of 10 nm to 70 nm, preferably 30 nm to 70 nm, more preferably 40 nm to 65 nm; and / or A composition according to any one of the preceding claims, having an average diameter of from 10 nm to 50 nm, preferably from 20 nm to 50 nm, more preferably from 30 nm to 40 nm, as measured by CryoTEM.

12. 10. The composition of any one of the preceding claims, wherein the liposome composition comprises phosphate buffered saline at a pH of about 6.

8.

13. 10. The composition of any one of the preceding claims, wherein the liposome composition comprises at least one of a pharmaceutically acceptable additive, carrier, excipient, and diluent.

14. 10. The composition of any one of the preceding claims, wherein the method for treating Parkinson's disease (PD) is selected from reducing tremor, increasing physical movement, increasing walking speed, improving walking ability, improving movement control, and improving speech changes in Parkinson's disease (PD) patients.

15. 1. A liposome composition for use in a method for treating Parkinson's disease (PD), the liposome composition comprising sphingomyelin in a lipid bilayer and a therapeutically effective amount of monosialotetrahexosylganglioside (GM1), wherein the treatment is at least partial reversal of Parkinson's disease (PD), and the treatment optionally comprises a dosing regimen in which a therapeutically effective dose of the liposome composition is administered in a primary administration mode at most every four days with at least three days between each administration, preferably at most every six days with at least five days between each administration, and most preferably at most every seven days with at least six days between each administration.

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