Methods for preventing, treating, and / or diagnosing neurodegenerative diseases

Abstract

The present invention relates to pharmaceutical compositions and diagnostic methods for neurodegenerative diseases, particularly Parkinson's disease. The invention includes compositions comprising anti-Clostridium tetani agents such as tetanus vaccines, anti-tetanus immunoglobulins, and antibiotics. Methods for diagnosis and monitoring Parkinson's disease comprising detecting C. tetani-derived nucleic acids, proteins, or analogues thereof, in biological specimens, are further provided.

Description

METHODS FOR PREVENTING, TREATING, AND / OR DIAGNOSINGNEURODEGENERATIVE DISEASESFIELD OF INVENTION

[0001] The present disclosure pertains to the field of medical sciences and biotechnology, specifically focusing on compositions and methods for the prevention, treatment, diagnosis, and monitoring of neurodegenerative diseases, including but not limited to Parkinson’s disease, by utilizing anti- Clostridium tetani agents and diagnostic biomolecular techniques.BACKGROUND OF THE INVENTION

[0002] Parkinson’s disease (PD) is a neurodegenerative disorder affecting over six million individuals worldwide. PD is characterized by the progressive loss of neurons throughout the peripheral autonomic and central nervous systems. Current diagnosis of PD is based on the presence of rest tremor, bradykinesia, rigidity and postural response abnormalities which are tightly associated with dopaminergic loss. The etiology of PD is poorly understood, with aging, genetic and environmental factors contributing to risk.

[0003] Currently, exercise is the only intervention consistently shown to slow PD progression. No approved therapy directly targets the underlying pathophysiological mechanism of the disease. Several treatment strategies are available to manage symptoms and to potentially slow disease progression. Pharmacological treatments include levodopa, which replenishes dopamine levels, dopamine agonists (e.g., pramipexole, ropinirole) that mimic dopamine, MAO-B inhibitors (e.g., selegiline, rasagiline) and COMT inhibitors (e.g., entacapone) that prolong dopamine action, amantadine, which reduces dyskinesias, anticholinergics, which help control tremors, and adenosine A2A receptor antagonists (e.g., istradefylline) that assist with motor fluctuations. Surgical interventions, such as deep brain stimulation (DBS) targeting the subthalamic nucleus or globus pallidus, provide motor improvement for advanced cases. Disease-modifying strategies, including a-synuclein-targeting antibodies and iron chelation therapy (deferiprone), are under investigation to slow neurodegeneration. While these therapies may improve quality of life, they are not directed to the basic mechanism of the disease and can cause side effects such as motorcomplications, cognitive impairment, psychiatric symptoms, nausea, and cardiovascular issues, necessitating individualized treatment plans.

[0004] Current Parkinson's disease treatments rely on symptomatic relief and clinical assessments for detection but do not address disease progression or underlying mechanisms. There is a need for therapies that modulate pathophysiology, preventive strategies for at-risk individuals, and diagnostic techniques enabling earlier detection. Improvements in these areas may enhance patient outcomes and mitigate the disease's impact.SUMMARY OF THE INVENTION

[0005] The invention addresses a significant unmet need in the field of neurodegenerative diseases. It relates to pharmaceutical compositions and diagnostic methods for neurodegenerative diseases, particularly Parkinson's disease. Uniquely, the invention establishes arAi-Clostridium tetani agents — such as tetanus toxoid vaccines, anti-tetanus immunoglobulins, and related antimicrobials — as preventive and / or disease-modifying therapies for Parkinson’s disease and potentially other neurodegenerative conditions. The invention also discloses the use of molecular or immunohistochemical detection of C. tetani -deriv ed toxins, or analogues thereof, in biological specimens as a diagnostic and / or monitoring approach for disease onset and progression.

[0006] Various objects of the invention are hereby presented in a non-limiting manner:

[0007] According to a first aspect there is provided a pharmaceutical composition comprising a therapeutically effective amount of an arAi-Clostridium tetani (C. tetani) agent and a pharmaceutically acceptable carrier, for use in the prevention and / or treatment of a neurodegenerative disease in a subject in need thereof.

[0008] In some embodiments, the anti-C. tetani agent is a tetanus vaccine or anti- tetanus immunoglobulins.

[0009] In some embodiments, the anti-C. tetani agent is a tetanus vaccine comprising tetanus toxoid.

[0010] In some embodiments, the tetanus vaccine further comprises diphtheria toxoid, pertussis toxoid, or both.

[0011] In some embodiments, the anti-C. tetani agent is anti- tetanus immunoglobulins administered via at least one route selected from the group consisting of intramuscular, intravenous, subcutaneous, intrathecal, intranasal, oral, intrapulmonary, and any combination thereof.

[0012] In some embodiments, the anti-C. tetani agent is an antibiotic therapy selected from the group consisting of benzathine penicillin, cefadroxil, erythromycin, roxithromycin, ofloxacin, levofloxacin, and any combination thereof.

[0013] In some embodiments, the anti-C tetani agent is selected from the group consisting of dipyridamole, eplerenone, verapamil, beclometasone, evolocumab, raloxifene, and any combination thereof.

[0014] In some embodiments, the neurodegenerative disease is selected from the group consisting of Parkinson’s disease, Alzheimer’s disease, multiple sclerosis, amyotrophic lateral sclerosis (ALS), Huntington’s disease, frontotemporal dementia (FTD), Lewy body dementia, multiple system atrophy, progressive supranuclear palsy, corticobasal degeneration, chronic traumatic encephalopathy, spinocerebellar ataxias, Friedreich’s ataxia, spinal muscular atrophy, prion diseases including Creutzfeldt- Jakob disease, fatal familial insomnia, and Gerstmann-Straussler- Scheinker syndrome, Wilson’s disease, Niemann-Pick disease type C, metachromatic leukodystrophy, adrenoleukodystrophy, Batten disease, hereditary spastic paraplegia, and Pelizaeus-Merzbacher disease.

[0015] In some embodiments, the neurodegenerative disease is Parkinson’s disease.

[0016] According to another aspect, there is provided a method for diagnosing Parkinson’s disease in a subject in need thereof, the method comprising detecting C. tetani- m . nucleic acid or protein in a specimen from said subject.

[0017] According to another aspect, there is provided a kit for diagnosing Parkinson’s disease in a subject, the kit comprising: (a) at least one reagent for detecting Clostridium tetani- m& nucleic acid or protein in a specimen obtained from the subject; and (b) instructions for using the kit to diagnose Parkinson’s disease based on the detection of the C. tetani- m . nucleic acid or protein.

[0018] In some embodiments, the level of said C. tetawz-derived nucleic acid or protein is above a predetermined threshold.

[0019] According to another aspect, there is provided a method for monitoring the progression and / or severity of Parkinson’s disease in a subject afflicted with Parkinson’s disease, the method comprising determining the level of C. tetani-Aeri eA nucleic acid or protein in a specimen from the subject and comparing its level to a previously determined level of C. tetani- m . nucleic acid or protein in identical specimen from the same subject.

[0020] In some embodiments, the C. tetani- m . protein is selected from the group consisting of tetanospasmin (tetanus toxin) protein, tetanolysin, a peptide degraded from said proteins, or an analogue thereof having at least 80% homology to said tetanospasmin or tetanolysin.

[0021] In some embodiments, the C. tetani- m& nucleic acid encodes tetanospasmin or tetanolysin or is an analogue having at least 80% homology thereto.

[0022] In some embodiments, the specimen is selected from the group consisting of blood, serum, plasma, cerebrospinal fluid, saliva, mucus, nasal content, stool, sweat, tears, and any combination thereof.

[0023] In some embodiments, detecting is via a PCR detection test of immunohistochemistry-based assay.BRIEF DESCRIPTION OF THE DRAWINGS

[0024] The following detailed description of embodiments of the presently disclosed subject matter refers to accompanying drawings:

[0025] Figures 1A-1B depicts forest plot (Fig. 1A) and contingency tables including regression models (Fig. 1A and Fig. IB), of Tetanus Diphteria (TD) vaccinations according to its timing with regards to the index date;

[0026] Figures 2A-2F demonstrate severity score kernel density plots and regressions;

[0027] Figures 3A-3H illustrate dot plots representing the dose-response relationship correlated with the acquisition of specified antimicrobial agents and food supplements; and,

[0028] Figures 4A-4F depict dot plots that illustrate the correlation between Parkinson's disease (PD) and the quantity of purchases for specific substances: dipyridamole (Figure 4A), eplerenone (Figure 4B), verapamil (Figure 4C), beclometasone (Figure 4D), evolocumab (Figure 4E), and raloxifene (Figure 4F).DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Methods of treating and / or preventing neurodegenerative diseases

[0029] The present invention is based, at least in part, on the surprising finding that anti-clostridium therapy protects against the development of a neurodegenerative disease, such as Parkinson’s disease.

[0030] Clostridium is a genus of anaerobic, spore-forming, Gram-positive bacteria. In some embodiments, the Clostridium is pathogenic Clostridium. In some embodiments, the Clostridium is a neurotoxinproducing clostridia. In some embodiments, the clostridium disclosed herein produces a neurotoxin. In some embodiments, the clostridium is Clostridium tetani.

[0031] Several neurotoxin-producing clostridia are known, including Clostridium tetani, C. botulinum, C. baratii, and C. butyricum. In some embodiments, it was unexpectedly found that a specific therapy against Clostridium tetani is effective in preventing and or treating the neurodegenerative disease.

[0032] According to a first aspect, there is provided a pharmaceutical composition comprising a therapeutically effective amount of an avAi-Clostridium tetani (C. tetani) agent and a pharmaceutically acceptable carrier, for use in the prevention of a neurodegenerative disease in a subject in need thereof.

[0033] According to another aspect, there is provided a pharmaceutical composition comprising a therapeutically effective amount of an anti-C. tetani agent and a pharmaceutically acceptable carrier, for use in the treatment of a neurodegenerative disease in a subject in need thereof.

[0034] In some embodiments, there is provided a method of treating a neurodegenerative disease in a subject in need thereof, the method comprising administering a therapeutically effective amount of an anti-C. tetani agent to a subject afflicted with the neurodegenerative disease.

[0035] In some embodiments, there is provided a method of preventing a neurodegenerative disease in a subject in need thereof, the method comprising detecting a subject suspectable to develop a neurodegenerative disease and administering a therapeutically effective amount of an anti-C. tetani agent.

[0036] As used herein, the term “treatment of a neurodegenerative disease” refers to any therapeutic intervention, approach, or method intended to alleviate, reduce, or manage the clinical symptoms of a neurodegenerative disorder, slow or halt disease progression, or modify or correct theunderlying molecular, cellular, or pathophysiological mechanisms contributing to the disease. The term “prevention of a neurodegenerative disease” refers to any intervention, strategy, or agent that delays the onset, reduces the risk, or lowers the likelihood of developing a neurodegenerative condition, including measures that interrupt or attenuate early pathological changes before the clinical manifestation of the disease.

[0037] As used herein, the term " arAi-Clostridium tetani agent" or "anti-C. tetani agent" refers to any substance that acts to neutralize or inhibit the activity of Clostridium tetani or its secreted toxin(s). Such agents may include, but are not limited to, antibodies, vaccines, antibiotics, and other therapeutics with similar mechanisms of action.

[0038] As used herein, the term "therapeutically effective amount" refers to a quantity of a pharmaceutical composition sufficient to achieve a desired therapeutic or prophylactic effect in the treatment or prevention of a specific disease or condition in a patient in need thereof. The exact amount will vary depending on multiple factors, including the nature and severity of the condition being treated, the patient's age, weight, general health status, and the specific pharmacological properties of the active compound being administered.

[0039] The term "pharmaceutically acceptable carrier" refers to an inert, non- toxic substance that is combined with an active pharmaceutical ingredient to facilitate its administration, improve stability, or enhance its delivery to a target site. Suitable carriers may include, but are not limited to, solvents, dispersion media, coatings, absorption-modifying agents, isotonic solutions, and controlled-release matrices. The choice of carrier depends on the intended route of administration and the physicochemical characteristics of the active compound. For example, parenteral formulations may include saline, buffered solutions, or dextrose, while oral formulations may utilize binders, fillers, or encapsulating agents. In the context of tetanus vaccination, commonly used carriers include aluminum- based adjuvants such as aluminum hydroxide or aluminum phosphate. These adjuvants are not merely carriers; they serve a dual role by enhancing the magnitude and duration of the immune response to the tetanus toxoid. In addition to adjuvants, tetanus vaccines may also include stabilizers and preservatives to ensure product stability, prolong shelf life, and maintain immunogenic efficacy throughout the storage period. Regarding tetanus immunoglobulins, the pharmaceutically acceptable carrier may include isotonic solutions such asnormal saline or buffered saline. The solution must be free of pyrogens and particulate matter, ensuring the sterility and safety of the administration. The immunoglobulins may be formulated with excipients to adjust pH, enhance solubility, and ensure proper osmolarity for the intended delivery route, be it intramuscular or intravenous.

[0040] Clostridium tetani is known to secrete the neurotoxin tetanospasmin (or TeNT) and the hemolysin tetanolysin. In some embodiments, the "ar i-Clostridium tetani agent" targets the neurotoxin tetanospasmin (TeNT).

[0041] Tetanospasmin (TeNT) is a single polypeptide with a molecular weight of approximately 150 kDa that is later cleaved into two chains — a heavy chain (100 kDa) and a light chain (50 kDa) — connected by a disulfide bond. It belongs to the A-B type exotoxin family, where the A (active) subunit is the light chain (L) with enzymatic activity, and the B (binding) subunit is the heavy chain responsible for cell binding and internalization. The light chain functions as a zinc-dependent endopeptidase that cleaves synaptobrevin (VAMP), a vesicle-associated SNARE protein essential for neurotransmitter release. As a result, the release of the inhibitory neurotransmitters GABA and glycine from interneurons in the central nervous system, is blocked. The heavy chain consists of two domains; the N-terminal domain (HN) the facilitates translocation of the light chain into the cytosol, and the C-terminal domain (HC) that binds specifically to neuronal receptors, particularly gangliosides and glycoproteins on motor neurons, enabling uptake by endocytosis and retrograde transport to the spinal cord.

[0042] In some embodiments, the "ax i-Clostridium tetani agent" disclosed herein is any agent that is capable of inhibiting and / or neutralizing the neurotoxin TeNT, including its binding to neuronal receptors, its translocation into the cytosol, or its downstream activity, e.g., cleavage of synaptobrevin. In some embodiments, the " arAi-Clostridium tetani agent" disclosed herein is an agent that targets the light chain of TeNT. In some embodiments, the "axdi-Clostridium tetani agent" is an agent that targets the heavy chain of TeNT. In some embodiments, it was unexpectedly found that there is a beneficial effect for targeting both the light chain and the heavy chain of TeNT. In some embodiments, the " ardi-Clostridium tetani agent" targets the N-terminal of the heavy chain. In some embodiments, the " arAi-Clostridium tetani agent" targets the C-terminal of the heavy chain. In some embodiments, the " arAi-Clostridium tetani agent" targets both the N-terminaland the C-terminal of the heavy chain. In some embodiments, the " axA\-Clostridium tetani agent" targets the full-lengh TeNT. In some embodiments, the "ax i-Clostridium tetani agent" disclosed herein is any agent that actively induces the level of anti-TeNT antibodies against the full-lengh TeNT.

[0043] In some embodiments, the anti-C. tetani agent is a tetanus vaccine comprising tetanus toxoid. Preparation of the tetanus toxoid can be via chemical inactivation of the toxigenic strains of Clostridium tetani. For example, the toxic strains can be grown in liquid media, purified, then treated with formaldehyde to take away the pathogenic properties. Following purification and sterilization, tetanus toxoid can be further combined with an adjuvant, e.g., aluminum or calcium salts.

[0044] The present invention is further based on the unexpected finding that there is a beneficial effect for an arAi-Clostridium tetani agent combined with an agent targeting diphtheria toxin, in preventing and / or treating the neurodegenerative disease. In some embodiments, the beneficial effect is a synergistic effect. In some embodiments, a synergistic effect was observed when tetanus toxoid was combined with diphtheria or pertussis antigens, resulting in enhanced neuroprotection compared to individual administration. In some embodiments, there is a beneficial effect for the arAi-Clostridium tetani agent combined with an agent targeting diphtheria toxin, an agent targeting pertussis toxin, or with both.

[0045] In some embodiments, there is provided a pharmaceutical composition comprising a therapeutically effective amount of tetanus toxoid and diphtheria toxoid for use in the treatment of a neurodegenerative disease in a subject in need thereof. In some embodiments, the pharmaceutical composition for use disclosed herein comprises tetanus toxoid and pertussis antigens. In some embodiments, the pharmaceutical composition disclosed herein comprises tetanus toxoid, diphtheria toxoid, and pertussis antigens.

[0046] The tetanus toxoid is an FDA-approved vaccination given alone or in conjunction with other vaccines. The toxoid antigen is usually used in combination with diphtheria and pertussis antigens in a vaccination form. The DTaP (diphtheria, tetanus, and acellular pertussis) vaccine is a 0.5 mL dose, and it is given intramuscularly (from Rabadi T, Brady MF. Tetanus Toxoid. [Updated 2023 May 1], In: StatPearls [Internet], Treasure Island (FL): StatPearls Publishing; 2025 Jan-. Availablefrom: htps: / / www.ncbi.nlm.nih.gov / books / NBK557415 / ). DTaP and Tdap (reduced diphtheria and pertussis for adolescents and adults) are both combination vaccines against diphtheria, tetanus, and pertussis. The "a" indicates that the pertussis toxoids are acellular, while the lower-case "d" and "p" in "Tdap" indicate smaller concentrations of diphtheria toxoids and pertussis antigens. Other tetanus toxoid combinations include DT (diphtheria and tetanus, for children), and Td (tetanus and reduced diphtheria, for adults). Acellular pertussis vaccines contain pertussis toxoid (PT), filamentous hemagglutinin (FHA), and pertactin (69K). The tetanus toxoid content varies by formulation: pediatric DTaP vaccines typically contain >20 IU of tetanus toxoid per 0.5 mb dose, while adult Td and Tdap formulations contain >5 IU per 0.5 mL. The vaccines can be adjuvanted with aluminum salts (e.g., aluminum hydroxide or aluminum phosphate) to enhance the immune response, but newer vaccines may explore other options. Administration may be by intramuscular injection, usually into the deltoid or anterolateral thigh. A primary vaccination series consists of three doses, followed by booster doses every 10 years to maintain protective antibody levels. For example, US8623380B2 discloses a diphtheria, tetanus and pertussis vaccine comprising a low dose of each of diphtheria toxoid (D), tetanus toxoid (T), pertussis toxin (PT), filamentous haemagglutinin (FHA) and pertactin (69K).

[0047] It is noted that, according to some embodiments, the tetanus vaccine for use disclosed herein may be based on the full-length holotoxin that has been chemically inactivated, e.g., using formaldehyde. This inactivation destroys the toxin’s enzymatic activity while preserving its structure sufficiently to elicit a robust immune response. As a result, the body generates neutralizing antibodies that recognize conformational epitopes across the entire toxin molecule, including both heavy and light chain domains.

[0048] In some embodiments, the pharmaceutical composition for use disclosed herein comprises mRNA- based vaccine against tetanus toxin. For example, the pharmaceutical composition may comprise a multivalent mRNA-DTP vaccine, as described in Wolf MA et al., Multivalent mRNA-DTP vaccines are immunogenic and provide protection from Bordetella pertussis challenge in mice. NPJ Vaccines. 2024 Jun 10;9(l ): 103, herein incorporated reference by its entirety.

[0049] In some embodiments, the pharmaceutical composition for use disclosed herein comprises antitetanus immunoglobulins or Tetanus Immune Globulin (TIG).

[0050] As used herein, the term Tetanus Immune Globulin (TIG) refers to antibodies against TeNT. TIG is usually used both as a prophylactic measure in patients with incomplete tetanus vaccination or as part of the treatment regimen for active tetanus. In some settings, equine-derived antitoxin may be used as an alternative, although it carries a higher risk of hypersensitivity reactions.

[0051] It is understood that the antibodies in TIG may target the binding domain of the c terminal of the heavy chain of TeNT (HC), responsible for the toxin binding to neuronal receptors, as well as the translocation (HN) and enzymatic (LC) domains, which are involved in the toxic action inside cells. The epitopes may include both conformational and linear epitopes across the toxin. In some embodiments, the pharmaceutical composition for use comprises polyclonal antibodies, monoclonal antibodies, or both.

[0052] In some embodiments, the method disclosed herein comprises administering the arAi-Clostridium tetani agent via at least one route selected from the group consisting of intramuscular, intravenous, subcutaneous, intrathecal, intranasal, oral, intrapulmonary, or any combination thereof.

[0053] In some embodiments, the anti-C. tetani agent may comprise small molecule inhibitors that block the translocation or enzymatic activity of TeNT (Zanetti G, Mattar ei A, Lista F, Rossetto O, Montecucco C, Pirazzini M. Novel Small Molecule Inhibitors That Prevent the Neuroparalysis of Tetanus Neurotoxin. Pharmaceuticals (Basel). 2021 Nov 8;14(11):1134, herein incorporated by reference in its entirety).

[0054] In some embodiments, the anti-C. tetani agent comprises antibiotic therapy employed to eliminate C. tetani bacteria and halt further toxin production. Metronidazole is known to be the antibiotic of choice due to its efficacy and lower risk of neurotoxicity compared to older agents. Penicillin G remains a viable alternative, although it is associated with a higher risk of seizure activity. Other antibiotics such as doxycycline, clindamycin, or erythromycin may be used in specific cases, particularly in patients with allergies or contraindications to first-line treatments.

[0055] In some embodiments, the anti-C. tetani agent comprises antibiotic therapy selected from the group consisting of benzathine penicillin, cefadroxil, erythromycin, roxithromycin, ofloxacin, levofloxacin, and any combination thereof.

[0056] According to another aspect there is provided a method for preventing and / or treating a neurodegenerative disease in a subject in need thereof, the method comprising administering atherapeutically effective amount of at least one substance selected from the group consisting of dipyridamole, eplerenone, verapamil, beclometasone, evolocumab, raloxifene, and any combination thereof.

[0057] Dipyridamole, which is known for its antiplatelet and vasodilatory effects, also influences the uptake of adenosine into cells, leading to increased extracellular adenosine levels. Adenosine has neuroprotective properties, including the modulation of neurotransmitter release and reduction of neuronal excitability. Elevated adenosine could therefore protect neurons from the excitotoxic effects of the tetanus toxin.

[0058] Eplerenone is a selective aldosterone receptor antagonist used primarily as a potassium-sparing diuretic. In the context of C. tetani, its protective role might involve the modulation of fluid and electrolyte balance in neuronal tissues, potentially affecting the environment necessary for the neurotoxin’s transport or action. By altering ion transport and cellular hydration, eplerenone could hypothetically impact the ability of the toxin to propagate through neuronal cells.

[0059] Verapamil is a calcium channel blocker that primarily affects cardiac muscle and blood vessels, but it also impacts neuronal calcium channels. By inhibiting these channels, Verapamil reduces the influx of calcium into neurons, which is crucial for neurotransmitter release and neuronal excitability. This reduction in calcium mitigates the synaptic dysfunction caused by the tetanus toxin, which relies on calcium-dependent processes in neurons.

[0060] Beclometasone is a corticosteroid that reduces inflammation by inhibiting multiple inflammatory cytokines and agents. In the setting of tetanus, inflammation can exacerbate neuronal damage. By reducing the inflammatory response, beclometasone lessens the secondary damage caused by the immune response to the infection and toxin.

[0061] Evolocumab is a monoclonal antibody that inhibits Proprotein convertase subtilisin kexin type 9 (PCSK9), a protein that degrades LDL receptors. While its primary use is to lower LDL cholesterol, the mechanism by which it could protect against tetanus toxin might involve the modulation of lipid metabolism and membrane composition in neurons, potentially affecting the toxin's ability to interact with neuronal membranes and enter nerve cells.

[0062] Raloxifene is a selective estrogen receptor modulator (SERM) that exhibits estrogen-like effects on bone and lipid metabolism but acts as an anti-estrogen in breast and uterine tissues. Estrogenhas neuroprotective effects, including the modulation of neurotransmitter systems and enhancement of neuronal survival. Raloxifene might mimic these effects in the nervous system, potentially reducing the susceptibility of neurons to toxin-induced damage.

[0063] In some embodiments, the neurodegenerative disease is selected from the group consisting of Parkinson’s disease, Alzheimer’s disease, multiple sclerosis, amyotrophic lateral sclerosis (ALS), Huntington’s disease, frontotemporal dementia (FTD), Lewy body dementia, multiple system atrophy (MSA), progressive supranuclear palsy (PSP), corticobasal degeneration (CBD), chronic traumatic encephalopathy, spinocerebellar ataxias, Friedreich’s ataxia, spinal muscular atrophy, prion diseases including Creutzfeldt-Jakob disease, fatal familial insomnia, and Gerstmann- Straussler-Scheinker syndrome, Wilson’s disease, Niemann-Pick disease type C, metachromatic leukodystrophy, adrenoleukodystrophy, Batten disease, hereditary spastic paraplegia, and Pelizaeus-Merzbacher disease.

[0064] In some embodiments, the neurodegenerative disease comprises Parkinson’s disease (PD) or atypical parkinsonian disorders.

[0065] Atypical parkinsonian disorders, also known as Parkinson-plus syndromes, are a group of neurodegenerative diseases that share some clinical features with Parkinson’s disease (PD) — such as bradykinesia and rigidity — but differ in underlying pathology, symptom profile, progression, and response to treatment. These include multiple system atrophy (MSA), characterized by autonomic dysfunction and cerebellar or parkinsonian features; progressive supranuclear palsy (PSP), marked by early postural instability, vertical gaze palsy, and axial rigidity; corticobasal degeneration (CBD), which presents with asymmetric motor symptoms, limb apraxia, and cortical sensory loss; and dementia with Lewy bodies (DLB), which combines parkinsonism with early cognitive decline, visual hallucinations, and fluctuating attention. Unlike PD, these disorders typically respond poorly to levodopa, show more rapid progression, and often involve early onset of non-motor symptoms such as cognitive impairment, autonomic failure, or eye movement abnormalities. Diagnosis is based on clinical assessment, supported by neuroimaging and, increasingly, investigational biomarkers.

[0066] In some embodiments, the neurodegenerative disease is PD.

[0067] In some embodiments, there is provided a method of treating PD in a subject in need thereof, the method comprising administering a therapeutically effective amount of an anti-C. tetani agent to a subject afflicted with PD.

[0068] In some embodiments, the method comprising administering a therapeutically effective amount of tetanus toxoid to a subject afflicted with PD.

[0069] In some embodiments, the method comprising administering a therapeutically effective amount of at least one substance selected from the group consisting of dipyridamole, eplerenone, verapamil, beclometasone, evolocumab, raloxifene, or any combination thereof, to a subject afflicted with PD.

[0070] In some embodiments, there is provided a method of preventing PD in a subject in need thereof, the method comprising detecting a subject suspectable to develop PD and administering a therapeutically effective amount of the anti-C. tetani agent.

[0071] The risk of PD is known to be increased among family members, although most cases are sporadic and not directly inherited. Having a first-degree relative (parent or sibling) with PD increases a person’s risk approximately 2-3 times compared to the general population. This increased risk is partly due to shared genetic factors and possibly shared environmental exposures.

[0072] Certain genetic mutations are known to increase the risk of PD, especially in early-onset or familial forms. Mutations in genes such as LRRK2, GBA1, SNCA, PARK2 (parkin), PINK1, and DJ-1 have been linked to PD. Among these, LRRK2 and GBA1 mutations are the most common in familial and even some sporadic cases.

[0073] Early diagnosis in high-risk individuals is an area of active research and may involve olfactory testing, as loss of smell is an early sign, REM sleep behavior disorder (RBD) screening, neuroimaging such as DaTscan to detect dopamine transporter loss, and detection of emerging biomarkers. Misfolded a-synuclein aggregates are a hallmark of PD, primarily found in Lewy bodies within brain tissue, and can be detected in various biofluids. In cerebrospinal fluid (CSF), total a-synuclein levels are often reduced, while detection in blood or plasma remains less reliable. Research is exploring skin or salivary gland biopsies, and seed amplification assays like RT-QuIC are promising for identifying pathological a-synuclein in CSF. Additionally, dopamine metabolites such as HVA (homovanillic acid) in CSF may be decreased, reflecting dopamine loss in PD.Neurofilament light chain (NfL) is elevated in atypical parkinsonian disorders, though not significantly in PD, and may help differentiate PD from other conditions. Lysosomal markers, including mutations in GBA1 and LRRK2, are known genetic risk factors, and related protein markers are under investigation in blood or CSF. Elevated inflammatory markers, such as cytokines and proteins, have been observed in PD patients, and metabolomic and proteomic signatures are being explored to identify complex biomarker patterns in blood, CSF, or urine as part of ongoing research.Methods of diagnosing Parkinson 's Disease

[0074] According to another aspect, a method for diagnosing Parkinson’s disease (PD) in a subject is provided, which involves detecting C. lelani-derwed nucleic acid or protein in a specimen from the subject.

[0075] In some embodiments, the level of C. tetani-derwed nucleic acid or protein is higher than a predetermined threshold.

[0076] In certain embodiments, the predetermined threshold is established by comparing the level of C. lelani-derwed nucleic acid or protein in the subject's specimen with that in a control specimen, which may come from a healthy subject or a subject affected by other neurodegenerative diseases.

[0077] According to another aspect, there is provided a method for monitoring the progression and / or severity of PD in a subject afflicted with PD, the method comprising determining the level of C. tetani-derwed nucleic acid or protein in a specimen from the subject and comparing its level to a previously determined level of C. tetani-derwed nucleic acid or protein in identical specimen from the same subject.

[0078] In some embodiments, the method comprises determining the level of C. tetani-derwed protein. In some embodiments, the C. tetani-derwed protein is selected from the group consisting of tetanospasmin (tetanus toxin) protein, tetanolysin, a peptide degraded from these proteins, or an analogue thereof having at least 80% homology to tetanospasmin or tetanolysin. In some embodiments, the C. tetani-derwed protein is tetanospasmin, or an analogue therefore having at least 80% homology to tetanospasmin. In some embodiments, the analogue protein has at least 80%, at least 85%, at least 90%, at least 95%, or at least 98% homology to tetanospasmin. Each possibility represents a separate embodiment of the present invention.

[0079] The amino acid sequence of tetanospasmin is highly conserved among different strains of C. tetani, with only minor variations observed. The complete sequence is documented in the UniProt database under the accession number P04958 (https: / / www.uniprot.org / uniprotkb / P04958 / entry).

[0080] In some embodiments, the method comprises determining the level of a nucleic acid encoding a Clostridium tetani-demed protein. In certain embodiments, the nucleic acid encodes tetanospasmin and / or tetanolysin; in specific embodiments, it encodes tetanospasmin. The nucleotide sequence of the tetanospasmin (TeNT) gene is available in multiple database entries, with notable differences between them. For example, GenBank accession Ml 8816.1 is an early submission from 1988 that represents the standalone tent gene (3861 bp) from an unspecified C. tetani strain. It was manually annotated and widely used in early molecular biology research and cloning (Eisel U, Jarausch W, Goretzki K, Henschen A, Engels JW, Niemann H. Tetanus toxin gene: nucleotide sequence and expression in Escherichia coli. EMBO J. 1986;5(10):2495- 2502. https: / / www.ncbi.nlm.nih.gov / nuccore / M18816). In contrast, RefSeq NC_004557.1 corresponds to the fully annotated pE88 plasmid from C. tetani strain E88, which includes the tent gene within a ~74 kb plasmid sequence (Bruggemann H, et al. The genome sequence of Clostridium tetani, the causative agent of tetanus disease. Proc Natl Acad Sci U S A. 2003 Dec 9; 100(23): 13113-8. doi: 10.1073 / pnas.1832525100. https : / / www. ncbi. nlm. nih. gov / nuccore / N C_004557.)

[0081] In some embodiments, the method comprises determining the level of a nucleic acid analogue having at least 80% sequence homology to the tetanospasmin gene. In specific embodiments, the analogue nucleic acid has at least 80%, 85%, 90%, 95%, or 98% homology to the tetanospasmin gene. Each degree of homology represents a separate embodiment of the present invention.

[0082] In some embodiments, the specimen is selected from the group consisting of blood, serum, plasma, cerebrospinal fluid (CSF), saliva, mucus, nasal content, stool, sweat, tears, or any combination thereof. In some embodiments, the specimen comprises CSF. In some embodiments, the specimen is CSF.

[0083] Diagnostic tools for detecting Clostridium tetani or its toxins include both molecular and immunoassay-based methods. PCR detection kits such as the Clostridium tetani Detection Kit (kitpcr.com) and the genesig® Advanced Kit (Primerdesign) target the tetanus toxin gene usingreal-time PCR technology. For antibody-based detection, ELISA kits like the Anti-Tetanus Toxoid IgG ELISA Kit (DRG International), Tetanus IgG ELISA Kit (DEIA10378) and Tetanus Toxin IgG ELISA Kit (DEIA1794) (Creative Diagnostics) offer sensitive quantification of anti-tetanus antibodies in serum or plasma. Rapid diagnostic tools such as the SD BIOLINE™ Tetanus Test (Maxanim) provide quick, qualitative results for IgG / IgM detection in blood samples. Additionally, specific antibodies against tetanus toxin are available for use in immunohistochemistry-based assays (Thermo Fisher Scientific), enabling detection of the toxin protein in tissue samples.EXAMPLESExample 1 - Materials and MethodsStudy design

[0084] This study was conducted as an observational analysis in Leumit Health Services (LHS), one of the four national health providers in Israel, providing comprehensive healthcare services to approximately 720,000 members. All Israeli citizens are entitled for comprehensive health insurance and receive a standardized package of health services and medications, as defined by the national “Health Basket” committee. LHS operates a centralized electronic health record (EHR) system, with over two decades of meticulously maintained information on patient demographics, medical diagnoses, healthcare encounters, laboratory test results, and records of prescribed and purchased medications. Diagnoses are documented during medical encounters by the treating physicians using the International Classification of Diseases, Ninth Revision (ICD-9). Diagnoses can be marked as chronic when they pertain to a chronic condition, and these can be updated or closed by the treating physicians during subsequent patient encounters. The reliability of these chronic diagnosis records in Leumit registry has been previously validated, demonstrating high accuracy.

[0085] Eligibility for inclusion in the study was defined as any past or current LHS member with at least five years of LHS membership between years 2003 and 2023. Data extraction was carried out from the LHS central data warehouse in February 2024, encompassing diagnoses, results of laboratory tests, and medication purchases recorded up to December 31, 2023.Cohort definition

[0086] The study cohort comprises eligible PD patients diagnosed for the first time between 45 and 75, alongside a control group matched at a 5: 1 ratio, of individuals with no documented PD. PD patients were identified by the presence of ICD-9 coded 332 diagnosis “Parkinson’s Disease” in the EHR, if recorded by a neurologist or a movement disorder specialist, or recorded by any physician, if accompanied with purchase of Antiparkinsonian medications over a period of more than six months. In order to avoid confusion with overlapping conditions, individuals with a diagnosis indicative of secondary parkinsonism, schizophrenia, pituitary adenoma, restless legs syndrome, or cerebrovascular accident were excluded from the study, as well of individuals with prior purchase of an antipsychotic medication susceptible to induce parkinsonism.

[0087] Controls were precisely matched to the PD patients based on gender, socio-economic status category, and the year of initial enrollment in Leumit Health Services (LHS). For each PD patient, five control individuals were chosen who met these matching criteria and whose birth dates were closest to that of the PD, ensuring no individual was duplicated within the cohort.Diphtheria-Tetanus toxoid vaccinations

[0088] The inventors looked for anti-tetanus vaccinations in the medical history of patients from the study cohort, using both vaccine purchase recorded by LHS pharmacies, and vaccine administration records documented in the EHRs. Most anti-tetanus vaccinations performed in LHS for patients in the cohort were with DT IMOVAX from Sanofi -Pasteur, adult dose, 0.5ml.Data preparation

[0089] Data were extracted from electronic health records and prepared for analyses using scripts developed by Leumit Research Institute in Python 3.11 with Pandas library and T-SQL queries. Prior to analysis patients’ data were deidentified and the patient ID number replaced with an identifier internal to the study. Graphs of this study were produced in Python with seaborn and matplotlib libraries.Statistical analysis

[0090] Statistical analysis was performed in R version 4.3. Unless specified otherwise, Fisher’s exact test was used to compare categorical variables and the two samples t-test to compare continuous variables across groups. Conditional logistic models were fit to assess the association of vaccination timing and covariates across matched groups. Linear regression models were fit toY1assess the relationship between linear variables such as disease severity and relative disease severity and explanatory variables. Pearson correlation analyses were performed to assess doseresponse relationship.Machine Learning Model

[0091] Gradient boosting model for predicting disease activity score was built with lightgbm python library following exploration of model parameters space performed with FLAML / AutoML using 5-fold cross-validation.Example 2 - ResultsThe study cohort

[0092] The inventors performed a large-scale observational study in Leumit Health Services (LHS), a nationwide health organization in Israel with over 22 years of centrally maintained electronic health records (EHR). Using rigorous criteria, they selected 1446 patients who received a PD diagnosis between the ages of 45 and 75, avoiding the rare cases occurring in very young age (<45), where disease is heavily influenced by genetic or environmental factors, and older patients (>75), where non-specific motor symptoms make the clinical diagnosis of PD less reliable. They used the earliest diagnosis date or antiparkinsonian medication purchase as the index date, and selected 7230 control individuals matching in a 5:1 ratio the PD patients. Control individuals, were assigned the same index dates as their respective cases, so that the same depth of recorded EHR history was available in the two groups.

[0093] Table 1 presents the demographic and clinical comparison of the two at the index date. The age, gender and socioeconomic distribution of the two groups are very similar. In both groups 55.6% were male, with an average age of 65.7 ± 7.2. Clinical characteristics also appear to be similar, with the notable exception of smoking status, with PD patients being less likely to be active smokers: Odds Ratio for current smoking (OR=0.591; P<0.001), consistent with literature.Table 1. Demographic and clinical characteristics of the study cohort at index dateVaccination is associated with decreased risk of PD occurrence

[0094] The last line of Table 1 displays a striking difference between the groups for TD vaccination status: only 1.52% of patients with PD had a record of vaccination before the index date, compared to 2.99% in the control group (OR=0.50, P=0.001).

[0095] A characteristic feature of a disease occurring because of waning antibody protection is a progressive increase in the disease rate with time elapsing since vaccination. Therefore, the relation between time elapsed since last vaccination and PD risk has been assessed.

[0096] PD risk in vaccinated patients is associated with time elapsed since vaccination

[0097] Figs. 1A-1C displays comparisons between the case and control groups according to the timing of vaccination. As the forest plot shows clearly, there is a time-dependent protection effect of the TD vaccination. The odds-ratio for PD occurring within two years after last TD vaccination is 0.00 (P=0.006), within 5 years, it is 0.17 (P=0.003). The odds increase to 0.26 between 5 to 10 years (P=0.004). Between 10 and 15 years post- vaccination, there is still a trend for protection, but the odds difference loses statistically significance. After 15 years post- vaccination the trend is for increased risk, not statistically significant.

[0098] If not getting a TD vaccination was to be a confounder of some early symptoms of PD (e.g. people with early signs of PD, before diagnosis, may decrease their activity and hence reduce opportunities to get wounded, and subsequently get vaccination), then one would expect that this pattern would continue and even amplify after the index date. But this does not occur: we observe an opposite trend, of increased odds for PD among those who got a vaccine after the index date (OR=1.33, P=0.39). This reversing time relationship strongly indicates that lack of TD vaccination is unlikely to be a confounder for early PD.

[0099] Having shown that PD occurrence is highly associated with time elapsed since last TD vaccination, we proceeded to verify, in the few patients who were diagnosed with PD after vaccination, or who were vaccinated once PD was diagnosed, whether the disease course was affected by vaccination. Vaccinated PD patients have slower rates of disease progression

[0100] In order to follow the disease course, the inventors developed a method to accurately assess disease severity during the follow-up period. Parkinson’s disease is characterized by progressive course of motor and functional deterioration as shown by Hoehn & Yahr’s clinical staging of the laterality and axial symptoms (Hoehn, M.M.; Yahr, M.D. Parkinsonism. Neurology 1967, 17, 427.), and by the Unified Parkinson’s Disease Rating Scale (UPDRS) (Ramaker, C.; Marinus, J.; Stiggelbout, A.M.; van Hilten, B.J. Systematic Evaluation of Rating Scales for Impairment and Disability in Parkinson’s Disease. Mov. Disord. 2002, 17, 867-876.) Symptoms are typically treated by anti-PD medications that are increased over time as the disease progresses in dosage and potency.

[0101] Having records of medications purchased by each patient, the inventors could use the annual medication consumption to train a machine learning model, that would assess disease severity for each patient and year of disease. For this purpose, a gradient boosting model was trained by cross- validation folds, utilizing yearly medication consumption per catalog entry as training variables, together with the gender of the patient (since disease course is affected by gender), to estimate time elapsed since disease onset, set as the target variable. The output of the model is a PD severity score expressed in a scale analogous to years of disease (e.g. a patient with a severity score of 6 in a given year has purchased medications that are typically used by a PD patient in the 6th year of the disease). Using this model, the inventors computed severity scores for 8,793 PD patient years of disease, of which 201 were from patients with prior vaccination record.

[0102] Fig. 2A displays a kernel density plot of these scores vs. the actual disease duration, along with a linear regression line. The calculated severity scores correlate very nicely with the actual year of the disease. The Pearson correlation coefficient, r is 0.443, with a P-value under 10-200. Of note, a quadratic regression model (a polynomial model of degree 2) was slightly more informative, exhibiting a lower Akaike Information Criterion (AIC) and a higher R2 compared to the linear model (see Fig. 2B).

[0103] Fig. 2C displays the severity scores over time, according to vaccination status. Disease severity in vaccinated patients is significantly lower than in non-vaccinated patients. Linear regression models show vaccination status is significantly associated with reduced severity, both in a univariable model (PO.OOl), and in a multivariable model adjusted for age, gender and smoking status (P<0.001). Here again the quadratic regression is more informative, so a quadratic model is used to plot disease severity according to time elapsed since PD diagnosis. Fig. 2D shows that, even though disease severity was roughly similar at disease onset, disease progression was significantly slower in the vaccinated group. The regression curve shows that the average severity in vaccinated patients at the seventh year of the disease was similar to the average severity score of unvaccinated patients who were at the third year of the disease.

[0104] It was further found out, that even among PD patients, disease course was less severe postvaccination than in PD patients with no vaccination record.Disease progression is associated to time elapsed since vaccination

[0105] The inventors have surprisingly found that PD progresses more slowly in patients who have been vaccinated than in those who did not. A further indication that TD vaccine affects PD progression would be to find, focusing on the vaccinated patients, an effect that is more pronounced when closer to the date of vaccination, even after correcting for time since diagnosis. For this analysis, which requires comparison of patients who have experienced varying durations of the disease, “relative PD severity scores” were calculated, which are obtained by subtracting the mean severity score of patients at the same disease year from the severity scores.

[0106] Fig. 2E displays these relative PD severity scores according to time since diagnosis, divided by vaccination status, showing once again that vaccinated individuals have lower disease severity (P<0.001). Fig. 2F follows the relative disease severity of vaccinated patients according to time elapsed since last vaccination. The relative disease severity decrease is inversely correlated with time elapsed since vaccination (P=0.03).Disease severity is affected by antimicrobial treatments

[0107] If clostridia present in the patients’ microbiome were actually involved in PD pathology, then PD disease severity would be affected by antimicrobial treatments that kill these bacteria. Havingcalculated PD disease severity scores, the inventors can use them to assess whether antimicrobial medications are actually associated with changes in disease severity.

[0108] For this analysis, forty classes of medications were identified for which varying consumption rates were observed between PD patients and control patients in the years prior to the index date, which is a potential signal for an effect on disease occurrence. For each of these classes, and for each year of disease in a PD patient, the inventors calculated a variable that reflects whether medications of the class were purchased by the patient over the three preceding years. Starting with a multivariable regression model that includes age, gender, and smoking status, a stepwise approach was employed to incrementally select the 10 medication classes most strongly associated with the relative PD severity scores. Table 2 presents the results of this regression. Not surprisingly, the tetanus toxoid vaccine had the strongest effect on disease severity, purchase of the vaccine in the preceding three years significantly reduced the relative disease severity (-1.11, P=0.0003).Table 2. Multivariable regression model examining the impact of medication purchased in the three preceding years on relative PD disease severity

[0109] Several antibiotic agents displayed a substantial and significant effect in reducing disease severity: benzathine penicillin, which is an intramuscular formulation of penicillin, displayed a significantreducing effect on disease severity (-0.69, P=0.003). Benzathine penicillin is the classical treatment for tetanus (Smith, J.W.G. Penicillin in Prevention of Tetanus. Br. Med. J. 1964, 2, 1293), its sustained release formulation is likely to deplete the clostridial population producing the toxin, explaining its beneficial effect on disease severity. Likewise, Cefadroxil, a first generation cephalosporine structurally close to penicillin displayed a significant decreasing effect on disease severity (-1.02, P=0.003). Two macrolide antibiotics also displayed a beneficial effect on disease severity: erythromycin (-0.74, P=0.0002) and roxithromycin (-0.13, P=0.008), as well as two compounds of the fluoroquinolone family: ofloxacin (-0.45, P<0.001), and levofloxacin (-0.22, P=0.026).

[0110] On the other hand, Clindamycin is associated with a significant and substantial increase in disease severity (+0.40, P=0.002). Clindamycin is primarily used to treat infections caused by susceptible anaerobic bacteria, but it is notorious for its risk of causing Clostridium difficile colitis, when Clostridium bacteria inherently resistant to clindamycin colonize the human colon (Duffy, C.R.; Huang, Y.; Andrikopoulou, M.; Stern-Ascher, C.N.; Wright, J.D.; Goffman, D.; D’Alton, M.E.; Friedman, A.M. Clindamycin, Gentamicin, and Risk of Clostridium Difficile Infection and Acute Kidney Injury During Delivery Hospitalizations. Obstet. Gynecol. 2020, 135, 59-67). The observed effect of increasing severity of PD is consistent with a competitive advantage granted by this antibiotic to clostridia, over susceptible anaerobes present in the microbiome, enabling proliferation of C. tetani in a manner similar to C. difficile proliferation following Clindamycin treatment.

[0111] Interestingly, the inventors also unexpectedly found strong apposing effects for two laxative agents commonly used to treat constipation in PD patients. Macrogol (polyethylene glycol), was significantly associated with increased disease severity (+0.34, P0.001), while ispaghula (Psyllium), a dietary fiber, was associated with decreased disease severity (-0.42, P=0.0004). Since these two compounds primary affect the gut environment, these opposite effects are strongly suggestive of a pivotal role played by an organism which resides in the intestinal microbiome.

[0112] Then it was tested whether a dose-response relationship could be detected, by looking for a correlation between the number of purchases by each patient during the three preceding years and the relative PD severity score. Figs. 3A-3H display scatter plots and correlation analyses for theidentified compounds. For most compounds the inventors found a significant dose-response relationship strengthening the direction of the association detected by the regression. Since the regression was based only on a binary variable for each compound, and did not account for the number of purchases, a significant dose-response relationship provides an independent corroboration of the effect detected for these medications.

[0113] Having identified C. tetani as the causative agent of Parkinson’s Disease, the inventors further identified, through an innovative algorithm they have developed, six molecules that display strong neuroprotective activity against neuronal damage present in Parkinson’s Disease. For each of these molecules; dipyridamole, eplerenone, verapamil, beclometasone, evolocumab, raloxifene, both a strong reducing effect on disease severity, and a dose-response relationship, have been identified (Figs. 4A-4F) Conclusion

[0114] This large-scale, real-life population study provides compelling evidence for the protective effect of diphtheria-tetanus toxoid vaccination on PD risk and its rate of progression. The significant associations observed between time elapsed since vaccination and both PD occurrence and rate of disease progression are all supporting the interpretation that the anti-tetanus vaccine provides protection against PD, and that this protection is waning over time. In addition, disease progression is significantly associated, in a dose-dependent manner, to antibiotics that affect clostridia growth negatively (beta-lactam, macrolides, fluroquinolones), or positively (clindamycin), strongly supporting the involvement of actual clostridium bacteria as the causative agent of disease.

Claims

CLAIMSWhat is claimed is:

1. A pharmaceutical composition comprising a therapeutically effective amount of an anti- Clostridium tetani (C. tetani) agent and a pharmaceutically acceptable carrier, for use in the prevention and / or treatment of a neurodegenerative disease in a subject in need thereof.

2. The pharmaceutical composition of claim 1, wherein said anti-C. tetani agent is a tetanus vaccine or anti- tetanus immunoglobulins.

3. The pharmaceutical composition of claim 2, wherein said anti-C. tetani agent is a tetanus vaccine comprising tetanus toxoid.

4. The pharmaceutical composition of claim 3, wherein said tetanus vaccine further comprises diphtheria toxoid, pertussis toxoid, or both.

5. The pharmaceutical composition of claim 2, wherein said anti-C. tetani agent is antitetanus immunoglobulins administered via at least one route selected from the group consisting of intramuscular, intravenous, subcutaneous, intrathecal, intranasal, oral, intrapulmonary, and any combination thereof.

6. The pharmaceutical composition of claim 2, wherein said anti-C. tetani agent is an antibiotic therapy selected from the group consisting of benzathine penicillin, cefadroxil, erythromycin, roxithromycin, ofloxacin, levofloxacin, and any combination thereof.

7. The pharmaceutical composition of claim 1, wherein said anti-C. tetani agent is selected from the group consisting of dipyridamole, eplerenone, verapamil, beclometasone, evolocumab, raloxifene, and any combination thereof.

8. The pharmaceutical composition of any one of claims 1 to 7, wherein said neurodegenerative disease is selected from the group consisting of Parkinson’s disease, Alzheimer’s disease, multiple sclerosis, amyotrophic lateral sclerosis (ALS), Huntington’s disease, frontotemporal dementia (FTD), Lewy body dementia, multiple system atrophy, progressive supranuclear palsy, corticobasal degeneration, chronic traumatic encephalopathy, spinocerebellar ataxias, Friedreich’s ataxia, spinal muscular atrophy, prion diseases including Creutzfeldt-Jakob disease, fatal familial insomnia, and Gerstmann-Straussler-Scheinker syndrome, Wilson’s disease, Niemann-Pick disease typeC, metachromatic leukodystrophy, adrenoleukodystrophy, Batten disease, hereditary spastic paraplegia, and Pelizaeus-Merzbacher disease.

9. The pharmaceutical composition of claim 8, wherein said neurodegenerative disease is Parkinson’s disease.

10. A method for diagnosing Parkinson’s disease in a subject in need thereof, the method comprising detecting C. lelani-derwed nucleic acid or protein in a specimen from said subject.

11. The method of claim 10, wherein the level of said C. lelani-derwed nucleic acid or protein is above a predetermined threshold.

12. A method for monitoring the progression and / or severity of Parkinson’s disease in a subject afflicted with Parkinson’s disease, the method comprising determining the level of C. lelani-derwed nucleic acid or protein in a specimen from said subject and comparing said level to a previously determined level of C. lelani-derwed nucleic acid or protein in said specimen from said subject.

13. The method of any one of claims 10 to 12, wherein said C. lelani-derwed protein is selected from the group consisting of tetanospasmin (tetanus toxin) protein, tetanolysin, a peptide degraded from said proteins, or an analogue thereof having at least 80% homology to said tetanospasmin or tetanolysin.

14. The method of any one of claims 10 to 13, wherein said C. tetani-der ed nucleic acid encodes tetanospasmin or tetanolysin or is an analogue having at least 80% homology thereto.

15. The method of any one of claims 10 to 14, wherein said specimen is selected from the group consisting of blood, serum, plasma, cerebrospinal fluid, saliva, mucus, nasal content, stool, sweat, tears, and any combination thereof.

16. The method of any one of claims 10 to 15, wherein said detecting is via a PCR detection test of immunohistochemistry-based assay.

17. A kit for diagnosing Parkinson’s disease in a subject, the kit comprising: (a) at least one reagent for detecting Clostridium tetani-demed nucleic acid or protein in aspecimen obtained from the subject; and (b) instructions for using the kit to diagnose Parkinson’s disease based on the detection of said C. ieiani-derwed nucleic acid or protein.

18. The kit of claim 17, wherein the reagent for detecting Clostridium tetani-derived nucleic acid comprises a set of primers and / or probes for polymerase chain reaction (PCR) amplification.

19. The kit of claim 17, wherein the reagent for detecting Clostridium tetani- derived protein comprises an antibody specific for said protein.

Images