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

Abstract

The present invention relates to a liposome composition for use in a method of treating Parkinson's disease. The present invention relates to a liposome composition comprising sphingomyelin and a therapeutically effective amount of monosialotetrahexosyl ganglioside (GM1) in a lipid bilayer wherein a therapeutically effective amount of the liposome composition is administered at most every 4 days in a predominant administration manner, at least three days between each administration; preferably, the medicine is administered every 6 days at most in a main administration mode, and the time interval between every two times of administration is at least 5 days; most preferably, the medicament is administered at most every 7 days in a main administration mode, and the interval between every two administration is at least 6 days.

Description

[0001] This invention relates to liposome compositions for methods of treating Parkinson's disease.

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

[0003] The most prominent signs and symptoms of Parkinson's disease (PD) occur when neurons in the basal ganglia, a brain region that controls movement, are damaged and / or die. Normally, these neurons produce an important brain chemical called dopamine. When neurons die or are damaged, they produce less dopamine, which causes motor problems associated with the disease.

[0004] Although there is no cure for Parkinson's disease (PD), medications, surgery, and other therapies can often alleviate some symptoms. Medications can help treat PD symptoms by increasing dopamine levels in the brain, by acting on other brain chemicals that transmit information between brain cells (such as neurotransmitters), or by helping to control nonmotor symptoms.

[0005] The primary treatment for Parkinson's disease is levodopa (or L-DOPA). Nerve cells use levodopa to produce dopamine to replenish the brain's reduced supply. Typically, levodopa is taken together 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 agitation—and reduces the amount of levodopa needed to improve symptoms.

[0006] Other medications used to treat symptoms of Parkinson's disease include dopamine agonists to stimulate the production of dopamine in the brain, enzyme inhibitors (such as MAO-B inhibitors and COMT inhibitors) to increase the amount of dopamine by slowing down the enzymes that break down dopamine in the brain, amantadine to help reduce involuntary movements, and anticholinergic drugs to reduce tremors and muscle stiffness.

[0007] Certain types of glycolipids have been shown to play a key role in the development and treatment of Parkinson's disease (PD). Glycolipids are lipids containing carbohydrates linked by glycosidic bonds. Their role in living cells is to maintain cell membrane stability and facilitate cell recognition. Sphingolipids are a subtype of glycolipids containing the amino alcohol sphingosine. They occur in the nervous system and other parts of the body, and they are involved in the metabolic and pathological changes associated with PD.

[0008] Ganglioside GM1 (monosialotetrahexosylganglioside, CAS: 37758-47-7) is a glycosphingolipid composed of branched-chain pentasaccharides. The branched-chain pentasaccharide consists of one sialic acid residue, two galactose residues, one N-acetylgalactosamine residue, and a glucose residue linked to N-stearoylsphingosine via a β-bond at the reduced end. GM1 possesses important physiological properties and influences neuronal plasticity and repair mechanisms, as well as the release of neurotrophic factors in the brain (VASQUES, JF et al., Gangliosides in nervous system development, regeneration, and pathologies. Neural Regen. Res. 2023, Vol. 18, No. 1, pp. 81-86). It is an amphiphilic endogenous molecule, abundant in the outer lobules of cell membranes in all mammals, and particularly enriched in the neuronal plasma membrane (accounting for 5% ± 10% of total lipid mass).

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

[0010] Higher gangliosides such as GM1 are present only in the late stages of organogenesis and fetal growth, suggesting their role in guiding the formation of a fully functional and young central nervous system (CNS). From a lipid-based perspective, the possibility of at least partially reintroducing this “young CNS” state 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, pp. 1643-1675).

[0011] GM1 belongs to the α-ganglioside family, and its biosynthesis occurs in the Golgi apparatus. Along with more complex gangliosides, GM1 is enriched in the presynaptic and postsynaptic membranes of synaptic terminals. Besides regulating nuclear calcium... 2＋In addition to homeostasis (through the high affinity binding of gangliosides to sodium / calcium exchangers at the nuclear envelope) and other cellular functions, GM1 also regulates neurotrophic activity by binding to receptors TrkA, TrkB, and GDNF (NEWBURN, EN et al., GM1 ganglioside enhancements Ret signaling in striatum. J. Neurochem. 2014, Vol. 130, No. 4, pp. 541-554).

[0012] The role of GM1 in the development and potential treatment of Parkinson's disease (PD) is of particular interest (CHOWDHURY, S. et al., The Key Role of GM1 Ganglioside in Parkinson's Disease. Biomolecules 2022, Vol. 12, No. 173, pp. 1-9). GM1 deficiency hinders the normal function of cells that depend on sufficient levels of this ganglioside, leading to their gradual degeneration and eventual death. Studies in humans have shown that GM1 deficiency may be a contributing factor to the development of idiopathic PD. A reduction in GM1 in the brains of deceased PD patients has been reported (WU, G. d et al., Deficiency of ganglioside GM1 correlates with Parkinson's disease in mice and humans. J. Neurosci. Res. 2012, Vol. 90, No. 10, pp. 1997-2008). GM1 levels in the brains of healthy individuals have been reported to decrease over time. However, this reduction is particularly significant 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, pp. 1-21).

[0013] Decreased GM1 levels were also observed in the pre-death cerebrospinal fluid (CSF) of PD patients (compared to age-matched controls), although this was not significant (a decrease of 17.4%). Similar serum analyses did indeed find a significant decrease in samples from PD patients (LEDEEN, R. et al., Systemic deficiency of GM1 ganglioside in Parkinson's disease tissues and its relation to the disease etiology. Glycoconj. J. 2022, Vol. 39, No. 1, pp. 75-82).

[0014] It has been reported that transgenic mice of the GM1 family, which are completely or partially lacking gangliosides, exhibit PD features according to behavioral and neuropathological criteria due to the disruption of the B4galnt1 gene (GM2 synthase-B4galnt1). These features 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 has improved these pathological CNS manifestations (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] Numerous clinical studies have explored the therapeutic potential of GM1 in the Parkinson's disease (PD) patient population. The first study was a small, pilot-scale investigation. This was an open-label study in which 10 PD patients received 1000 mg of GM1 via intravenous infusion after the last of their three baseline functional assessments, followed by self-administration of GM1 at a subcutaneous dose of 200 mg / day for 18 weeks. Under these conditions, GM1 gangliosides demonstrated to be safe and well-tolerated. No serious adverse events occurred, and no patients developed elevated anti-GM1 antibody titers (SCHNEIDER, JS et al., GM1 ganglioside treatment of Parkinson's disease: an open pilot study of safety and efficacy. Neurology 1995, Vol. 45, No. 6, pp. 1149-1154).

[0016] This study was followed by a larger, randomized, placebo-controlled trial involving 45 individuals with mild to moderate PD (using the same GM1 treatment regimen as described above). In this study, significant differences between groups in the Unified PD Rating Scale (UPDRS) motor score (p=0.0001) and activities of daily living (ADL) section (non-periodic assessment; p=0.04) were reported at week 16 (SCHNEIDER, JS et al., Parkinson's disease: improved function with GM1 ganglioside treatment in a randomized placebo-controlled study. Neurology 1998, Vol. 50, No. 6, pp. 1630-1636). In an open-label extended follow-up study, at the end of 5 years of GM1 use, the researchers concluded that "long-term use of GM1 in patients with PD is safe and provides certain clinical benefits." Participants typically had lower UPDRS motor scores (assessed during the actual defined “off” period) (SCHNEIDER, JS et al., GM1ganglioside in Parkinson's disease: Results of a five-year open study. J.Neurol. Sci. 2010, Vol. 292, No. 1, pp. 45-51).

[0017] These findings support the initiation of a larger-scale delayed start study. Of the 157 individuals screened, 77 were randomized to either an early start or delayed start group for GM1 treatment. The GM1 treatment regimen was the same as in previous studies. A separate comparison group of 17 individuals was used for comparison with standard care. At week 24, the motor scores of the UPDRS in the early start group significantly improved, while the scores in the delayed start group significantly worsened. The early start group also showed a sustained benefit at weeks 72 and 120 compared to the delayed start group. Both groups showed significant symptom deterioration during the washout period 2 years after GM1 treatment. The study demonstrates that long-term administration of GM1 is safe and well-tolerated, and it reports no increase in anti-GM1 antibody titers in participants (SCHNEIDER, JS et al., A Randomized, Controlled, Delayed Start Trial of GM1Ganglioside in Treated Parkinson's Disease Patients. J Neurol. Sci. 2013, Vol. 324, No. 1, pp. 140-148). Fifteen participants from the early initiation group and fourteen from the delayed initiation group volunteered for the PET(11)C-methylphenidate imaging study (eleven members from the comparison group also participated). Scans were performed on all three groups at baseline, week 24, and 1–2 years later. Results showed a significant slowing of binding loss in several striatal regions in the GM1-treatment group. In some cases, increased binding was detected after GM1 use (SCHNEIDER, JS et al., GM1 ganglioside in Parkinson's disease: Pilot study of effects on dopamine transporter binding. J Neurol. Sci. 2015, Vol. 356, No. 1, pp. 118–123).

[0018] One of the challenges in translating the therapeutic potential of GM1 from preclinical models to clinical applications is its ability to adequately reach the CNS when administered peripherally. CNS penetration is severely limited by the amphiphilic nature of GM1, which hinders pathways across the blood-brain barrier. Therefore, novel methods for delivering GM1 to the CNS are needed for clinical application.

[0019] Furthermore, despite promising signs in GM1 research, certain side effects (adverse events) still hinder its commercialization and need to be overcome. For example, pain, skin irritation, and severe hematoma at the injection site have occurred during GM1 treatment, as the most common route of administration for GM1 is subcutaneous.

[0020] To improve the bioavailability of GM1 in the CNS, liposomes have been proposed as vesicles for drug delivery in the treatment of PD (WO 2019122220 A1 (INNOMEDICA HOLDING AG) 27.06.2019, pp. 7-8). However, therapeutically effective formulations and dosing regimens do not yet exist in the prior art.

[0021] Therefore, one object of the present invention is to address these needs and provide improved treatment for PD. Specifically, the object of the present invention is to reduce the adverse events frequently encountered with frequent subcutaneous administration.

[0022] These problems have been solved by using liposomal GM1 formulations as medicines, which have the features according to the independent claims.

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

[0024] Liposome compositions are compositions containing liposomes in addition to other components, wherein one or more of the other components are encapsulated within liposomes. Liposomes are spherical vesicles having at least one lipid bilayer. Liposomes can also be multivesicular liposomes, where one 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 have the potential to provide controlled release of the encapsulated active pharmaceutical ingredient (API) over a prolonged period by limiting the concentration of the free component in the bloodstream and tissues, and to reduce the side effects of the encapsulated component. Liposomes can also modify tissue distribution and API uptake in a therapeutically advantageous manner and can increase the convenience of treatment by allowing for less frequent drug administration. For example, liposomes can deliver the encapsulated API directly to the disease site. The active ingredient can be released directly from the liposome at the treatment site. Therefore, lower doses of the active ingredient are required, thereby limiting side effects.

[0025] In cases of PD, liposomes containing a suitable lipid membrane composition enhance permeation of the blood-brain barrier. The blood-brain barrier comprises blood vessels that vascularize the CNS and tightly regulate the movement of ions, molecules, and cells between the blood and the CNS. This leads to increased deposition of APIs encapsulated in liposomes within the CNS.

[0026] The preparation of liposome compositions containing GM1 is described in the prior art (WO 2019122220 A1 (INNOMEDICA HOLDING AG) 27.06.2019). Specifically, the liposomes used in the liposome compositions of the present invention have a roundness greater than or equal to 0.95, particularly 0.98-1.00. The roundness of the liposomes in the formulation is determined by cryo-transmission electron microscopy (CryoTEM).

[0027] Lipids derived from the lipid membrane composition used to form the liposomes of the present invention are ubiquitous in healthy brain tissue. The liposomes of the present invention comprise sphingomyelin, which belongs to the group of phospholipids and sphingolipids. It accounts for approximately 10% of brain lipids. Sphingomyelin tends to have the highest concentration in the plasma membrane of biological cells, particularly in the outer lobules.

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

[0029] The API of this invention, GM1, is partially located in the membrane and partially in the aqueous core of the liposome. As previously mentioned, ganglioside GM1 is known in the art to play a significant role in the development and potential treatment of PD. According to the International Union of Pure and Applied Chemistry (IUPAC), the chemical name of 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)oxacyclohexane-2-yl]oxyoxygen Heterocyclohexane-2-yl]oxy-2-[(2R,3S,4R,5R,6R)-4,5-dihydroxy-6-[(E,2R,3S)-3-hydroxy-2-(eicosylamino)eicoseno-4-enoxy]-2-[(hydroxymethyl)oxetane-3-yl]oxy-3-hydroxy-6-[(hydroxymethyl)oxetane-4-yl]oxy-4-hydroxy-6-[(1R,2R)-1,2,3-trihydroxypropyl]oxetane-2-carboxylic acid. GM1 has the following structural formula:

[0030]

[0031] Formula 1

[0032] A therapeutically effective dose of monosialotetrahexosylganglioside (GM1) refers to the amount of GM1 that results in at least one of the following: prevention or delay of the onset of PD symptoms in a subject; improvement of PD symptoms in a subject; prevention or delay of the onset of PD signs in a subject; improvement of PD signs in a subject; or a combination thereof. As understood in the art, symptoms are felt or experienced, such as pain or dizziness. As understood in the art, signs are objective, observable indications of disease, injury, or abnormal physiological state that can be detected during physical examination, review of patient history, or diagnostic procedures. Signs and symptoms are not mutually exclusive.

[0033] These symptoms and signs include unintended or uncontrollable movements such as swaying, stiffness, and difficulty in balance and coordination, as well as difficulty walking and talking. The Movement Disorders Society Unified PD Rating Scale (MDS-UPDRS) is typically used to assess symptoms.

[0034] The efficacy of PD treatment is typically measured using the MDS-UPDRS system. This system comprises four parts: Part I assesses the non-motor aspects of daily living experiences using a questionnaire; Part II assesses the motor aspects of daily living experiences using a questionnaire; Part III conducts a motor examination during a clinical examination (without a questionnaire); and Part IV assesses treatment-induced motor complications using a questionnaire. A higher MDS-UPDRS value indicates a more pronounced disease.

[0035] In the context of this invention, dosage is a measured amount of a therapeutic agent taken once, wherein the measured amount may be administered over a period of time.

[0036] In the context of this invention, a therapeutically effective dose of a liposome composition is a dose of a liposome composition containing a certain amount of GM1 that results in at least one of prevention or delay of symptom onset; or improvement of PD symptoms in a subject.

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

[0038] According to the present invention, a dose of a liposomal composition containing GM1 is administered at most every 4 days via the primary route of administration, with an interval of at least 3 days between each administration; preferably, it is administered at most every 6 days via the primary route of administration, with an interval of at least 5 days between each administration; most preferably, it is administered at most every 7 days via the primary route of administration, with an interval of at least 6 days between each administration. For example, "administered at most every 7 days, with an interval of at least 6 days between each administration" means, for example, if a dose is administered on Monday, the next dose via the primary route of administration will be administered no earlier than the following Monday.

[0039] The primary route of administration may be any route of administration that a person skilled in the art would deem suitable for administering the dose over a reasonable period of time.

[0040] It has been found that liposome compositions containing sphingomyelin as a GM1 carrier system, as described in this invention, result in enhanced pharmacokinetic and therapeutic properties of the encapsulated GM1.

[0041] Unwilling to be bound by theory, the liposome composition according to the present invention increases the delivery of GM1 to the CNS because the liposome composition has higher penetration of the blood-brain barrier than non-liposome GM1. Furthermore, intracellular delivery of GM1 is also improved by encapsulating GM1 in the liposome composition. Non-liposome GM1 remains extracellular to a greater extent due to its lower cell membrane permeability. Therefore, the biodistribution of GM1 is advantageously altered with the liposome composition to increase drug deposition in the CNS.

[0042] Furthermore, not wanting to be bound by theory, it was found that the circulation of liposomal GM1 is prolonged compared to non-liposomal GM1 because liposomal GM1 is metabolized and secreted at a lower rate.

[0043] Due to its high permeability to both the blood-brain barrier and cell membranes, as well as its prolonged circulation time, GM1 requires a smaller amount to achieve the desired pharmacological effects when used in liposomal compositions compared to non-liposomal GM1. This, in turn, means that GM1 needs to be administered at a lower frequency.

[0044] As discussed earlier, when GM1 is administered by injection, particularly subcutaneous injection, and by other types of injection, severe pain, severe hematoma, and skin irritation at the injection site have been observed. It has been found that liposomal compositions enable smaller doses of GM1 per administration and / or lower dosing frequencies, leading to a reduction in the distribution of adverse events in PD patients. The result is less severe or even no pain, hematoma, and skin irritation.

[0045] Surprisingly, the liposome composition of the present invention was found to have a longer serum half-life in venous plasma than that of non-PEGylated (PEG: polyethylene glycol) liposome compositions containing sphingomyelin observed in normal practice. Non-PEGylated liposome compositions containing sphingomyelin for medical applications reported in the literature have serum half-lives 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 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 to prolong the half-life of liposomes in plasma is PEGylation of liposomes (PARK, H. et al., Evolution of drug delivery systems: From 1950 to 2020 and beyond. JCR 2022, Vol. 342, pp. 53-65). However, the liposome composition of the present invention does not require PEGylation of the liposomes and still exhibits a half-life as long as 30 h. This indicates that, using the liposome composition of the present invention, a half-life of a non-PEGylated liposome composition containing sphingomyelin is obtained that would not be expected by those skilled in the art. This results in longer time intervals between dosing administrations without significant loss of therapeutic effect between doses.

[0046] Specifically, it was found that for most treated PD patients, the liposomal GM1 composition needs to be administered at most every 7 days to achieve the desired therapeutic effect throughout the dosing interval. Initial findings suggest that, in some cases, more frequent dosing (e.g., every 4 or 6 days) is beneficial for maintaining therapeutic efficacy between doses. However, this still represents an improvement compared to existing dosing regimens, such as twice daily.

[0047] The advantages, improved efficacy, and reduction in adverse events will be shown in the Examples section below.

[0048] In a preferred embodiment, the composition is administered intravenously as the primary route of administration. It has been found that adverse events can be reduced not only by the smaller amounts of GM1 and lower dosing frequency required as described above, but also by using intravenous injection instead of conventional subcutaneous injection. This results in less pain, hematoma, and skin irritation in PD patients. Furthermore, intravenous administration leads to increased bioavailability compared to other forms of administration because, for example, the gastrointestinal barrier can be bypassed.

[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 a dose is administered within this timeframe compared to a shorter administration period. In the context of this invention, the 45 to 90-minute administration period refers to the first dose. For the second and third intravenously administered doses, even longer administration periods of 90 minutes to 10 hours are recommended to avoid adverse events such as back and neck pain and potential pseudoallergic reactions (e.g., complement activation-related pseudoallergic reactions (CARPA)).

[0050] In a preferred embodiment, administration via the primary route of administration is accompanied by administration via a secondary route of administration between doses of the primary route of administration, wherein the secondary route of administration is preferably oral administration. The pharmacological effect is increased by administering additional doses via the secondary route of administration between doses of the primary route of administration. For example, if the primary route of administration is weekly intravenous administration and the secondary route of administration is daily oral administration of the composition, high levels of GM1 in the body can be maintained. Simultaneously, with such an approach, limitations on quality of life are minimized. Subcutaneous injection of non-liposomal GM1 twice daily, as is common, leads to serious side effects. In contrast, weekly intravenous injection of liposomal GM1 with additional oral doses in between, which does not require the presence of a healthcare professional during oral administration, is significantly more beneficial to the patient and is associated with fewer side effects. Surprisingly, despite the lower total amount and frequency of GM1 administration, this results in better pharmacological outcomes due to the liposomal composition.

[0051] Preferably, the primary and / or secondary dosing regimens comprise equal or increased doses of the liposomal composition containing GM1. In PD patients experiencing such side effects, administering a smaller amount of the liposomal composition in an earlier dosing can counteract potential adverse events that frequently occur in the first few doses. Once fewer side effects are observed in PD patients, the dose can be increased accordingly for subsequent dosings. These side effects of the liposomal composition include back and neck pain, as well as pseudoallergic reactions (e.g., complement activation-related pseudoallergic reactions (CARPA)) or reactions of humoral origin. It should be noted that no clinically relevant adverse events have been observed in the dosing regimens corresponding to the present invention.

[0052] As an alternative to administering a smaller amount of the liposomal composition to mitigate adverse events during intravenous administration, the infusion rate can be adjusted so that a smaller amount of the liposomal composition flows into the body per unit time. Furthermore, both the dosage and rate of administration of the liposomal composition can be adjusted during the same administration process. The choice of dosage and rate of administration depends primarily on patient feedback. For example, if a patient complains of pain in the neck and / or back and / or near the injection site during administration, the dosage and / or rate of administration can be adjusted accordingly. Additionally, the infusion can be interrupted until the symptoms of the adverse event subside.

[0053] In the context of this invention, a dosing regimen is a timetable of therapeutic doses per unit time, including the time between doses, the amount of therapeutic agent per dose, and the time period for administering the dose, for example, via intravenous injection. Several dosing regimens using the same method of administration can be used consecutively to treat a patient, for example, if the time between doses changes. Furthermore, several dosing regimens can be used in parallel in cases involving different methods of administration.

[0054] Preferably, the primary route of administration includes a second dose of the liposome composition containing GM1 at a dose lower than the first dose, and additional third and subsequent doses.

[0055] In a preferred embodiment, the composition is administered regularly every 7 days in the primary administration route, with an interval of 6 days between each administration.

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

[0057] In another preferred embodiment, the liposome composition further comprises cholesterol, preferably a 1:1 molar ratio of sphingomyelin and cholesterol. Liposomes comprising sphingomyelin and cholesterol exhibit enhanced cycle life and CNS bioavailability. They possess improved pharmacokinetic and therapeutic properties. They are biocompatible and biodegradable. Although elevated cholesterol levels may be observed in some cases after administration of the liposomal cholesterol-containing composition, these elevated cholesterol levels will return to normal without any resulting adverse events.

[0058] In a preferred embodiment, a therapeutically effective amount of GM1 in a single dose of the liposomal composition is selected for the primary route of administration, such that it results in a venous plasma concentration of GM1 of 50 μg / ml to 1200 μg / ml, particularly 75 μg / ml to 600 μg / ml, and further particularly 100 μg / ml to 400 μg / ml, achieved within 1 h to 7 h, preferably 3 h to 5 h, and most preferably 4 h after the start of administration. This range of venous plasma concentrations of GM1 has been found to result in optimal pharmacological effects without causing serious side effects. It has been found that for such values ​​of venous plasma concentration of GM1, the GM1 concentration remains significantly higher than baseline even after 96 h (see the Examples section below). This confirms the expectation of a long-term circulating drug.

[0059] In 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 have 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) refers to DLS of samples with lipid concentrations ranging from 20 mg / ml to 30 mg / ml, diluted 1 / 19 in phosphate-buffered saline (PBS) or milliQ H2O to achieve an in-instrument attenuation factor 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 Zetasizer software (version 7.11) from Malvern. Particle size (hydrodynamic diameter) was determined using the Stokes-Einstein equation.

[0061]

[0062] Where k is the Boltzmann constant and T is the absolute temperature. D 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 an autocorrelation function using an appropriate algorithm. Cumulative analysis is a simple method for analyzing the autocorrelation function generated by DLS experiments and producing the average particle size (Z-ave) and polydispersity index (PDI). The calculations are defined in ISO 13321 (1996) and ISO 22412 (2008). The first-order result from the DLS experiments is the intensity distribution of the particle size. The intensity distribution is naturally weighted according to the scattering intensity. The particle size distribution is shown as a graph of the relative intensity of light scattered by the particles (on the Y-axis) versus the logarithmic interval of various particle size grades (on the X-axis). Measurements were performed using a transparent disposable zeta cell with a path length of 10 mm. Typically, but not necessarily, the liposomes in the compositions of this invention will fall within the size range measured by the method described.

[0063] "Measured by CryoTEM" refers to the examination of samples by cryotransmission electron microscopy (CryoTEM). Liposome samples were appropriately diluted, vitrified, and prepared on a grid (Formvar and Carbon) using an accelerating voltage of 200 kV. Images were acquired using a CryoTEM JEOL JEM-2100F, TVIPS TemCam F415MP camera at magnifications of 20,000x, 40,000x, and 80,000x. Particle identification and size determination were performed using semi-automatic image processing with Vironova analyzer software (Vironova, Sweden). In short, a series of random images at the same magnification were imported. Only liposome particles completely within the image boundaries and possessing a distinct membrane were detected. The sphericity, roundness, and unilamellarity of the identified objects were analyzed. All images were batch-processed using the same threshold and settings, accumulating more than 5 to 18 images per sample, corresponding to 1560 to 1178 analyzed particles (6 particles). The average value has a standard deviation of approximately 10 nm.

[0064] Typically, but not necessarily, the liposomes in the formulations of this invention will fall within the numerical range of sizes measured by both methods. The diameter size measured by CryoTEM is typically smaller than the diameter size measured by DLS.

[0065] In the context of this invention, the phrase "and / or" in relation to the measurement of liposome diameter should be understood as an inclusive segment. This means that it includes all liposome compositions, wherein liposomes typically, but not necessarily, meet any one or both of the conditions.

[0066] It has been successfully demonstrated that the liposomes of the present invention, with the stated composition and indicated size, are more stable than those known in the prior art. These smaller-diameter liposomes are less rapidly and to a lesser extent conditioning compared to their larger counterparts, and are less rapidly cleared by the reticuloendothelial system. Furthermore, larger liposomes are more likely to fuse or interact with other liposomes or particles.

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

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

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

[0070] In the context of this invention, an additive is a substance added to a composition along with an API to impart specific qualities to the composition. Additives may have very little or no therapeutic value, but are necessary in the preparation of a particular dosage form. Additives may be used for any one or any combination of the following purposes: providing bulk to the composition; promoting drug absorption or solubility and other pharmacokinetic considerations; assisting in the handling of APIs during manufacturing; providing stability and preventing denaturation.

[0071] In the context of this invention, a carrier (also known as a drug carrier or drug delivery medium) is a substrate used in drug delivery processes 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, an excipient is a substance formulated with an API, including for the purpose of long-term stability, filling a solid dosage form containing a small amount of the active ingredient, or imparting a therapeutic enhancement to the API, such as promoting drug absorption, reducing viscosity, or enhancing solubility or bioavailability.

[0073] In the context of this invention, a diluent (also known as a filler, diluent, or thinner) is a component in a pharmaceutical formulation that lacks pharmacological activity but is pharmaceutically necessary or desirable. It is particularly useful in increasing the volume of an effective drug that is too small to be manufactured or administered for the required dosage.

[0074] In a preferred embodiment, the method of treating PD is selected from reducing tremor, increasing body movement, improving walking speed, improving walking ability, improving motor control, and improving speech changes in patients with PD. The aforementioned complaints are common symptoms in PD patients. Treatment of affected areas of the CNS with the liposome composition according to the invention has shown that these symptoms can be significantly reduced when the severity of symptoms is assessed using the MDS-UPDRS system.

[0075] Another aspect of the invention relates to a liposomal composition for a method of treating Parkinson's disease (PD), said liposomal composition comprising sphingomyelin in a lipid bilayer and a therapeutically effective amount of monosialotetrahexosylganglioside (GM1), wherein the treatment is at least partially reversing PD, and wherein said treatment optionally comprises a dosing regimen in which a therapeutically effective dose of said liposomal composition is administered at most every 4 days as a primary dosing route, with an interval of at least 3 days between each administration; preferably, administration is administered at most every 6 days, with an interval of at least 5 days between each administration; most preferably, administration is administered at most every 7 days, with an interval of at least 6 days between each administration.

[0076] In the context of this invention, at least partial reversal of PD is at least partial reduction of at least one of the symptoms or signs of the disease, wherein at least one of the symptoms or signs is substantially no increase compared to baseline.

[0077] Surprisingly, it was found that treating PD patients with the liposome composition of the present invention over a 24-week period not only halted the negative progression of PD but also significantly reduced symptoms and signs, even when the liposome composition of the present invention was administered at 7-day intervals.

[0078] The present invention will be further explained through the following embodiments. These examples are not intended to limit the scope of the invention in any way.

[0079] The preparation of liposome compositions containing GM1 is described in the prior art (WO2019122220A1 (INNOMEDICA HOLDING AG) 27.06.2019). In particular, the liposomes used in the liposome compositions of the present invention have a roundness greater than or equal to 0.95, especially 0.98-1.00.

[0080] In the examples below, the liposome composition containing GM1 for intravenous administration may be referred to as “TLN” or “Talineuren”.

[0081] Figures 1a-1c Striatal levels of dopamine and its metabolites in MPTP-PD model mice: Dopamine levels (DA) determined by HPLC. Figure 1a ), 3,2-dihydroxyphenylacetic acid level (DOPAC, Figure 1b ) and high vanillic acid levels (HVA, Figure 1c n=5-10, bars represent mean and standard deviation, p-values ​​for one-way ANOVA treatment groups *<0.05, **<0.01;

[0082] Figure 2 Reverse rotation bar test performance in C9orf72 mice: Reverse rotation bar test was performed from P110 to P151 (15 mg / kg saline or Talineuren (TLN), intravenously, every other day, dashed box), n=5-7 / grp. Values ​​represent the mean and the standard error of the mean. Differences were assessed using a two-way ANOVA with Bonferroni multiple comparison test, ****p<0.0001;

[0083] Figure 3 Hematoxylin and eosin (H&E) and NADH staining were performed on the gastrocnemius muscle of C9orf72 mice at P170. Left: H&E staining of the gastrocnemius muscle. Black arrows indicate angled 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 animals / group. Samples were stained and imaged using the same settings.

[0084] Figure 4 Performance of SOD1-G93A mice in the reverse rotation bar test, starting at P110–P121 and P160–P170 (15 mg / kg saline or Talineuren ganglioside (TLN), intravenously, every other day, dashed box), n=4–6 mice / group. Values ​​represent the mean and the standard error of the mean. Differences were assessed using a two-way ANOVA with Bonferroni multiple comparison test, ****p<0.0001;

[0085] Figure 5a +5b: MDS-UPDRS movement in each patient (P1-P7; P9-P12) between baseline and assessment 8 weeks after treatment. Figure 5a ) and total ( Figure 5b Differences in scores. 5a: Changes in UPDRS motor score (“off” standard drug treatment for 12 hours) (R); 5b: Changes in UPDRS total score (S);

[0086] Figure 6a +6b: MDS-UPDRS movement in each patient (P1-P7; P9-P12) between baseline and assessment at 24 weeks post-conclusion. Figure 6a ) and total ( Figure 6b Differences in scores. 6a: Changes in UPDRS motor score (“off” standard drug therapy for 12 h) (R); 6b: Changes in UPDRS total score (S);

[0087] Figure 7 The changes in plasma GM1 concentration (U) over time (T) in all 9 patients in the dose consolidation group, and the mean value at each measurement time point;

[0088] Figure 8a +8b: TLN-DiI uptake in mouse brain cells: 8a: Substantia nigra after 48 h (TLN-DiI, iv, 30 mg / kg). DAPI (nuclear DNA, blue), DiI (dye, red). 8b: Motor cortex after 24 h (TLN-DiI, iv, 15 mg / kg). Scale bar = 50 μm, white arrows indicate DiI-positive cells;

[0089] Figure 9 GM1 levels (V) in the rat brain were measured by LC-MS / MS after once-daily intravenous administration of 12.3 mg / kg body weight of GM1 (free or in the form of Talineuren). (t-test, Welch correction, p-value <0.05), bars represent mean and standard deviation, n=5;

[0090] Figure 10 : Mean score (X) of the Parkinson's Disease Questionnaire-39 (PDQ-39) over time at baseline (time t=0 weeks (w)), 8 weeks after treatment, and 24 weeks after treatment; W: time in weeks (w).

[0091] Talineuren has been studied in preclinical models of many neurological diseases. Efficacy studies have been conducted in animal models of various movement disorders, such as the MPTP-mouse model of PD, and the C9orf72 and Sod1 mouse models of amyotrophic lateral sclerosis.

[0092] Example 1: Talineuren in the PD model

[0093] MPTP (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine) is a neurotoxin. ＋ The prodrug of PD causes permanent symptoms of PD by destroying dopaminergic neurons in the substantia nigra of the brain.

[0094] Studies in a mouse model of PD using MPTP (where dopaminergic neurodegeneration is induced by neurotoxins) yielded mixed results. MPTP was administered twice daily at 4-hour intervals, at a dose of 20 mg / kg in saline, intraperitoneally, for a total dose of 80 mg / kg. The administration volume of MPTP was 10 mL / kg, resulting in a pure MPTP active compound concentration of 1.6 mg / mL (after a reduction in the salt correction factor). MPTP administration began in the morning (8–9 AM).

[0095] Mice received Talineuren orally daily for 14 days (days 1-14). The administration volume was 5 ml / kg. On MPTP administration days (days 1-2), the compound was administered in the evening (6-8 PM). On day 3, the compound was administered in the afternoon (1-2 PM). From days 4-14, the compound was administered starting at 7 AM.

[0096] Figure 1 shows the determination of dopamine (DA) by high performance liquid chromatography (HPLC). Figure 1a ), 3,2-dihydroxyphenylacetic acid (DOPAC), Figure 1b ) and homovanillic acid (HVA, Figure 1c The striatal level was measured. Five to ten subjects were examined in each treatment group; bars represent the mean and standard deviation in wet tissue. Each group received the following treatments:

[0097] A: Fake treatment;

[0098] B: MPTP + PBS;

[0099] C: MPTP + BV (basal vesicles) 30 mg / kg, orally;

[0100] D: MPTP + GM1 30 mg / kg, intraperitoneally;

[0101] E: MPTP + GM1 30 mg / kg, orally;

[0102] F: MPTP + TLN 7.5 mg / kg, orally;

[0103] G: MPTP + TLN 15 mg / kg, orally; and

[0104] H: MPTP + TLN 30 mg / kg, orally

[0105] In the first study, oral administration of Talineuren at doses of 7.5 mg / kg (F) and 15 mg / kg (G) showed partial recovery of dopamine and dopamine metabolite levels in the substantia nigra.

[0106] Example 2: Talineuren in the ALS model

[0107] Amyotrophic lateral sclerosis (ALS) is a progressive neurological disease that affects nerve cells in the brain and spinal cord, leading to loss of muscle control.

[0108] To evaluate its potential therapeutic effects in ALS, two preclinical models with the most frequent mutations in familial ALS were used. In C9orf72 and SOD1 mouse models of amyotrophic lateral sclerosis (ALS), Talineuren gangliosides improved motor performance and reduced ER stress at the cellular level. Liposome uptake was observed in in vitro cultures of cortical neurons, but also in in vivo spinal motor neurons. More importantly, Talineuren ganglioside treatment in the C9orf72 transgenic model of ALS showed a significant effect on motor performance. Motor function and coordination were significantly improved not only when intravenous Talineuren treatment was initiated in the early stages of the disease but also when treatment was initiated in later stages.

[0109] Figure 2 This shows the performance of C9orf72 mice in the reverse rotating bar test. The test is used to assess motor coordination and balance in rodents. The apparatus consists of a motorized circular bar that rotates at a constant or increasing speed. The rodents naturally attempt to remain on the rotating bar to avoid falling onto the platform. Fall latency (L) is measured in seconds. Figure 2 The following curves are shown:

[0110] I: Wild-type mice injected with saline only;

[0111] J: Wild-type mice injected with Talineuren;

[0112] K: C9orf72- mice injected with saline only;

[0113] M: C9orf72- mice injected with Talineuren.

[0114] Treatment began on day 110 after birth (P110) and continued until day 151 (P151). Figure 2 (Dashed box in the image). When intravenous treatment was initiated and continued in the early stages of the disease, prolonged fall latency (L) approaching wild-type (WT) levels was observed in C9orf72 mice injected with Talineuren (M). This high fall latency (L) persisted until the end of P151 treatment. This effect remained highly significant throughout the treatment period. In a hang-string test performed on day 140 (P140), muscle strength was also improved in C9orf72 mice treated with Talineuren gangliosides compared to the C9orf72 saline group.

[0115] Microscopic observation of spinal cord sections supports these changes in the mouse's physical condition. Figure 3 As demonstrated by the downregulation of BiP (a marker of endoplasmic reticulum stress), Talineuren ganglioside treatment alleviates metabolic stress, a marker known to be involved in ALS progression.

[0116] PolyGA, a neuronal inclusion derived from the repetitive amplification of proteins in C9orf72 mutations, is abundant in ALS patients and detected in post-mortem tissues. PolyGA aggregation is associated with unfolded protein responses and triggers behavioral deficits through inflammation and protein sequestration, which may contribute 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, pp. 241-254). In Talineuren-treated animals, large PolyGA aggregates were significantly reduced. 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, pp. 521-534). In preclinical trials, H&E staining of the gastrocnemius muscle at P170 revealed angular fiber clusters and myofibril clusters with central nuclei in untreated C9orf72 mice, indicating myofibril degeneration and denervation, which was not observed in WT mice or C9orf72 mice treated with Talineuren (see [link to article]). Figure 3 (See left-hand image). Additionally, nicotinamide adenine dinucleotide (NADH) staining revealed type I and type II myofibrils in WT mice. In untreated C9orf72 mice, reduced myofibril structure indicated mitochondrial or sarcoplasmic reticulum abnormalities and neurogenic processes (see [link to image]). Figure 3 (See the image on the right). C9orf72 animals that received Talineuren almost all exhibited WT characteristics.

[0117] The therapeutic effect of intravenously administered Talineuren was also observed in a preclinical ALS model with SOD1 mutations. Figure 4 The following curves are shown:

[0118] N: Wild-type mice injected with saline only;

[0119] O: Wild-type mice injected with Talineuren;

[0120] P: C9orf72- mice injected with saline only;

[0121] Q: C9orf72 mice injected with Talineuren.

[0122] The two treatment intervals, P110 to P121 and P161 to P171, are in Figure 4 The following are marked with a dashed box. A 10-day treatment starting at day P110 postnatal resulted in improved spinner performance in mutant animals. Although the effect diminished over time, the treated animals maintained the improved performance. However, a second treatment interval at P160 did not enhance performance.

[0123] Example 3: Clinical Trial Results

[0124] A Phase I clinical trial was conducted as an open-label, single-dose escalation trial (n=3), followed by multiple doses at the maximum appropriate dose (n=9) (Neurologisches Institut Konolfingen; ClinicalTrials.govidentifier: NCT04976127). The primary objective was to demonstrate the safety of intravenous TLN in patients with Parkinson's disease. Secondary objectives were to determine the maximum appropriate dose based on safety profiles and preliminary efficacy, and to determine pharmacokinetic characteristics.

[0125] Dosage escalation

[0126] In all three PD patients, the dose was increased weekly to the maximum expected dose of 720 mg without any serious adverse events, dose delays, or dose changes. Therefore, the maximum appropriate dose for the dose consolidation phase of the trial was determined at the highest dose level (720 mg). This confirmed the expected good tolerability of the drug.

[0127] Dosage consolidation

[0128] Nine additional patients were enrolled in this trial and treated with a defined therapeutic dose for eight weeks. All nine patients completed Talineuren treatment for eight weeks. Serum blood samples were collected at 10 different time points on the first day of Talineuren administration and in the following days to determine the pharmacokinetics of GM1. All samples were analyzed, and the results are presented below.

[0129] Optional extension of treatment

[0130] All patients enrolled in this trial indicated a desire to continue weekly Talineuren treatment because they subjectively experienced benefits from it. Because Talineuren is generally well-tolerated, the trial was extended twice (Extension 1: 8 weeks; Extension 2: 16 weeks). This allowed patients to continue ultimately benefiting from Talineuren while collecting further safety data on the investigational drug with repeated 720 mg doses.

[0131] Safety / Tolerability

[0132] Treatment was generally well tolerated and no serious adverse events occurred. Several patients experienced CARPA (complement activation-related pseudohypersensory reaction; patients experienced neck pain and headache, which rapidly ceased as the infusion rate decreased), but this was not classified as a serious adverse event according to the safety definition of the clinical trial protocol. CARPA is a known phenomenon in investigational nanotechnology drugs, and all CARPA experienced in this study was completely reversible.

[0133] Efficacy after 8 weeks of treatment

[0134] The efficacy analysis after 8 weeks of treatment included data from all patients (11 of 12 patients) who were treated with weekly infusions of 720 mg of the active pharmaceutical ingredient GM1 ganglioside in the liposomal formulation of the present invention throughout the 8-week study. One patient (patient 8) received only 2 weeks of treatment and withdrew from the study.

[0135] For the measurement of therapeutic efficacy, two evaluation methods were considered:

[0136] 1) MDS-UPDRS motor score (Part III). Following the literature (SCHNEIDER, JS 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, pp. 140-148), evaluation was conducted in a "discontinued" medication state, meaning the patient had stopped their standard treatment for 12 hours. Evaluation was performed at two time points: the first was the baseline (BL) measurement before treatment initiation, and the second was one week after the final treatment during the final assessment (FA) at 8 weeks later.

[0137] 2) MDS-UPDRS Total Score. Measured as described in Part III of 1). Parts I, II, and IV correspond to the standard evaluation procedure for MDS-UPDRS using a questionnaire. Evaluations are performed during the first Talineuren treatment (BL) and one week after the last treatment (FA).

[0138] Table 1 shows the data points for the two considered efficacy endpoints (motor score in the "off" state and MDS-UPDRS total score) for each patient based on the trial database. Columns 3 and 6 show the differences between the baseline (BL) assessment and the final assessment (FA) at 8 weeks for each patient. The mean motor score decreased by 6.7 points, and the total score decreased by 12.2 points. The differences were statistically significant at the 1% level using a Wilcoxon signed-rank test for nonparametric paired samples. Clinically, this difference is also considered relevant, as the "minimum clinically important difference" (MCID) improved by approximately 4.3 to 5 points according to the literature.

[0139]

[0140] surface

[0141] Figure 5a and Figure 5b Differences in MDS-UPDRS motor scores (Part III) and total MDS-UPDRS scores between baseline (BL) and final assessment (FA) at 8 weeks are shown in descending order for each patient (P1–P7; P9–P12). Labels on the Y-axis indicate:

[0142] R: Changes in UPDRS exercise score ("off" standard drug treatment for 12 h);

[0143] S: Changes in the overall UPDRS score.

[0144] Efficacy after 24 weeks of treatment

[0145] Due to promising results after 8 weeks of treatment, treatment was continued for a total duration of 24 weeks. Efficacy analysis after 24 weeks of treatment included data from patients P1-P7 and P9-P12, who received weekly infusions of 720 mg of the active pharmaceutical ingredient GM1 ganglioside in the liposomal formulation of this invention.

[0146]

[0147] surface

[0148] Table 2 shows the data points for patients P1-P7 and P9-P12 on both considered efficacy endpoints (motor score and MDS-UPDRS total score in the "off" state). Columns 3 and 6 show the differences between the baseline (BL) assessment and the final assessment (FA) at 24 weeks for each patient. The mean motor score decreased by 7.5 points, and the total score decreased by 12.2 points. These results indicate a further improvement in the MDS-UPDRS motor score (Part III) at 24 weeks of total treatment compared to the results at 8 weeks of total treatment.

[0149] Figure 6a and Figure 6b Differences in MDS-UPDRS motor scores (Part III) and total MDS-UPDRS scores between baseline (BL) and final assessment (FA) at 24 weeks are shown in descending order for each patient (P1–P7; P9–P12). Labels on the Y-axis indicate:

[0150] R: Changes in UPDRS exercise score ("off" standard drug treatment for 12 h);

[0151] S: Changes in the overall UPDRS score.

[0152] Efficacy after 56 weeks of treatment

[0153] Similar to the efficacy analysis after 24 weeks of treatment, for the same patients except for P11, treatment lasted a total of 56 weeks. Again, patients received weekly infusions of 720 mg of the active pharmaceutical ingredient GM1 ganglioside in the liposomal formulation of this invention.

[0154]

[0155] surface

[0156] The data in Table 3 show that even after 56 weeks, these values ​​remained at a low level compared to the baseline.

[0157] Parkinson's Disease Questionnaire (PDQ-39)

[0158] In addition to using the Unified Parkinson's Disease Rating Scale (UPDRS) to determine the severity of Parkinson's disease, the Parkinson's Disease Questionnaire-39 (PDQ-39) was used to analyze the effectiveness of treatment. The questionnaire consists of 39 questions that assess the frequency with which individuals with Parkinson's disease experience difficulties in eight areas of daily living (dimensions): mobility, activities of daily living (ADL), emotional health, stigma, social support, cognition, communication, and physical discomfort. The total score for each area ranges from 0 (patients never have difficulty) to 100 (patients always have difficulty).

[0159] A questionnaire survey was conducted on the 12 patients P1-P12 at baseline (t=0), 8 weeks after treatment, and 24 weeks after treatment (patient P8 did not complete the questionnaire at 24 weeks). At each time point (w, t=0w, 8w, and 24w), the mean values ​​for the eight areas of daily living were calculated for each patient. Based on the results for the 12 patients, the mean, median, maximum, and minimum values ​​were calculated for each time point.

[0160] Figure 10 These values ​​(X) are shown as changing over time (W; in weeks (w)). The mean (circle) shows the PDQ-39 score decreasing from 20.1 at time 0 to 17.2 at 8 weeks, and then to 15.6 at 24 weeks. The median (triangle), maximum (square), and minimum (diamond) show the same trend over time. Particularly strong improvement was observed in the patient with the most severe initial score (patient 6: 33.9). The maximum score at 8 weeks (28.7) and the maximum score at 24 weeks (23.3) both belong to this patient.

[0161] These results indicate that significant improvements were observed in patients treated with the liposomal composition and dosage regimen according to the invention, not only in the MDS-UPDRS assessment procedure but also in the second assessment procedure (PDQ-39 scoring system).

[0162] Pharmacokinetics / Pharmacodynamics

[0163] Samples were collected from nine patients receiving weekly infusions of 720 mg Talineuren (dose consolidation portion) at 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).

[0164]

[0165] surface

[0166] Table 4 shows the time (t) for each of the nine patients to reach peak plasma concentration. 最大 ), highest plasma concentration (c 最大 ) and the half-life of GM1 in plasma (t 1 / 2 In addition, the mean, standard deviation (SD), minimum, median, maximum, coefficient of variance (CV%), geometric mean, and geometric mean of the coefficient of variance are shown. Figure 7 This shows the changes in plasma GM1 concentration over time for all nine patients, as well as the mean value at each measurement time point. The labels on the axes indicate:

[0167] T: Time since infusion began, in hours;

[0168] U: GM1 concentration in ng / ml.

[0169] GM1 was measured in all samples. Most patients reached their peak venous plasma concentrations after 4 hours. The infusion lasted 90 minutes, and no samples were taken at that time point. Therefore, peak plasma concentrations could be reached earlier. Significantly higher GM1 levels were still observed at 96 hours compared to baseline, confirming the expectation of long-term circulating drug use.

[0170] The mean half-life was slightly lower than expected, although a few individual cases were higher (patient 11), with a large standard deviation. With the currently measured mean half-life and a dosing frequency of only once a week, there may be no accumulation in plasma. Therefore, testing different dosing frequencies may be of interest to achieve steady-state conditions. The mean volume of distribution was relatively low and may indicate that GM1 is predominantly found in the blood. It may also mean that GM1 is widely distributed in the central nervous system rather than in other hydrophobic parts of the body.

[0171] Example 4: Brain Penetration

[0172] The delivery of Talineuren to the brain has been evaluated in mice. Fluorescently labeled Talineuren (TLN-DiI) was administered to mice, and dye accumulation was measured after 24 h or 48 h.

[0173] Figure 8a This shows the uptake of TLN-DiI in mouse brain substantia nigra cells after 48 h (TLN-DiI, iv, 30 mg / kg). Figure 8a The left image shows nuclear DNA (DAPI, blue) and DiI dye (red). Figure 8a The right image only shows the DiI dye (red). Figure 8b This shows TLN-DiI uptake in cells of the motor cortex of the brain 24 hours later (TLN-DiI, iv, 15 mg / kg). DiI signaling was detected in cells of the substantia nigra and motor cortex (white arrows).

[0174] In a study using CD-1 rats, the accumulation of the active pharmaceutical ingredient (GM1) of Talineuren in the rat brain was investigated. Each animal received intravenous administration of free (GM1) or liposomal GM1 (TLN) at a dose of 12.3 mg / kg body weight once daily for 4 consecutive days. The level of GM1 in the brain was quantified by liquid chromatography-mass spectrometry / mass spectrometry (LC-MS / MS) 24 h after the last administration.

[0175] Figure 9The study showed GM1 levels (V) in the rat brains four days after treatment. The levels were significantly increased in the group receiving Talineuren (liposomal GM1; TLN) compared to the group receiving free GM1. This indicates that GM1 is delivered to the brain more efficiently when inserted into liposomes.

Claims

1. A liposomal composition for treating Parkinson's disease (PD), the liposomal composition comprising sphingomyelin in a lipid bilayer and a therapeutically effective amount of monosialotetrahexosylganglioside (GM1), wherein the therapeutically effective dose of the liposomal composition is administered at most every 4 days via primary administration, with an interval of at least 3 days between administrations; preferably, administered at most every 6 days via primary administration, with an interval of at least 5 days between administrations; most preferably, administered at most every 7 days via primary administration, with an interval of at least 6 days between administrations.

2. The composition according to claim 1, wherein, The composition is administered intravenously via the primary route of administration.

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

4. The composition according to any one of the preceding claims, wherein, Dosing via the primary route of administration is accompanied by dosing via a secondary route of administration between doses of the primary route of administration, wherein the secondary route of administration is preferably oral administration.

5. The composition according to any one of the preceding claims, wherein, The primary and / or secondary administration routes include dosing regimens of equal or increased doses of the liposome composition containing GM1.

6. The composition according to any one of claims 1 to 4, wherein, The primary administration method includes administering a second dose of the GM1-containing liposome composition at a rate lower than the first dose, and / or administering the second dose at a flow rate lower than the first dose.

7. The composition according to any one of the preceding claims, wherein, The composition is administered regularly every 7 days in the primary route of administration, with an interval of 6 days between each administration.

8. The composition according to any one of the preceding claims, wherein, The therapeutically effective dose of the liposome composition is GM1 of 300 mg to 800 mg, preferably 600 mg to 750 mg, and most preferably about 720 mg.

9. The composition according to any one of the preceding claims, wherein, The liposome composition also contains cholesterol, preferably a sphingomyelin and cholesterol in a 1:1 molar ratio.

10. The composition according to any one of the preceding claims, wherein, The therapeutically effective amount of GM1 in a single dose of the liposomal composition is selected for the primary route of administration such that it results in a venous plasma concentration of GM1 of 50 μg / ml to 1200 μg / ml, particularly 75 μg / ml to 600 μg / ml, and even more particularly 100 μg / ml to 400 μg / ml, achieved within 1 h to 7 h after the start of administration, particularly within 3 h to 5 h, and even more particularly within 4 h.

11. The composition according to any one of the preceding claims, wherein, The liposomes of the liposome composition —Having 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 —Having an average diameter of 10 nm to 50 nm, preferably 20 nm to 50 nm, more preferably 30 nm to 40 nm, as determined by CryoTEM.

12. The composition according to any one of the preceding claims, wherein, The liposome composition comprises a phosphate-buffered saline solution with a pH of about 6.

8.

13. The composition according to any one of the preceding claims, wherein, The liposome composition comprises at least one of pharmaceutically acceptable additives, carriers, excipients, and diluents.

14. The composition according to any one of the preceding claims, wherein, The methods for treating Parkinson's disease (PD) are selected from those that reduce tremors, increase physical activity, improve walking speed, improve walking ability, improve motor control, and improve speech changes in patients with Parkinson's disease (PD).

15. A liposomal composition for a method of treating Parkinson's disease (PD), said liposomal composition comprising sphingomyelin in a lipid bilayer and a therapeutically effective amount of monosialotetrahexosylganglioside (GM1), wherein, The treatment is at least a partial reversal of Parkinson's disease (PD), and the treatment optionally includes a dosing regimen in which a therapeutically effective dose of the liposomal composition is administered at most every 4 days as the primary dosing route, with an interval of at least 3 days between each administration; preferably, administration is administered at most every 6 days, with an interval of at least 5 days between each administration; most preferably, administration is administered at most every 7 days, with an interval of at least 6 days between each administration.

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