Alkaline phosphatase for use in the treatment of neurodegenerative disorders

Abstract

The present invention relates to the treatment of neurodegenerative diseases, a group of chronic progressive disorders characterized by the gradual loss of neurons or neuronal function in discrete regions of the central nervous system (CNS). In particular, the present invention relates to alkaline phosphatase (AP) for use in the treatment or prevention of mammals suffering from or at risk of neurodegenerative disorders caused by reduced peroxisome proliferator-activated receptor gamma coactivator 1 (PGC1) activity.

Description

Technical Field

[0001] Specification The present invention relates to the treatment of a group of chronic progressive disorders characterized by neurodegenerative diseases, i.e., the gradual loss of neurons or neuronal function in separate regions of the central nervous system (CNS). Specifically, the present invention relates to alkaline phosphatase (AP) for use in the treatment or prevention of mammals suffering from or at risk of neurodegenerative disorders caused by reduced peroxisome proliferator-activated receptor gamma coactivator 1 (PGC1) activity.

Background Art

[0002] Neurodegenerative diseases are a group of chronic progressive disorders characterized by the gradual loss of neurons in separate regions of the central nervous system (CNS), and include, for example, Alzheimer's disease (AD), Parkinson's disease (PD), amyotrophic lateral sclerosis (ALS), multiple sclerosis (MS), and neurodegeneration after stroke. As used herein, frailty, an age-related syndrome, and progressive disorders associated with neurological deterioration are also included. Frailty is a comorbidity of diseases and disorders, in which the body gradually loses its inherent reserves and becomes vulnerable to dramatic and sudden changes in health, including neurodegeneration, triggered by seemingly minor events, such as mild infections or changes in medication or environment. Frailty is related to the aging process and is generally characterized by problems such as muscle weakness and fatigue. The mechanisms underlying their progressive nature remain unknown, although substantial evidence has described inflammatory mechanisms common to various neurodegenerative diseases. PD is a neurodegenerative disease for which there is currently no treatment. PD is characterized by a number of frequently occurring comorbidities (most of which are also seen in frailty), including reduced bone formation, reduced muscle mass, higher intestinal permeability, worsening of intestinal dysbiosis, higher blood-brain barrier (BBB) permeability, reduced spatial memory, increased low-grade inflammation, and reduced insulin sensitivity, in addition to the well-known motor symptoms of rigidity and tremor.

[0003] Parkinson's disease (PD) is the second most common neurodegenerative disease after Alzheimer's disease (AD) and is the most common motor disorder. Currently, approximately 2% of the population over 60 years of age are affected. Prominent clinical features include motor symptoms such as bradykinesia, tremors, rigidity, and postural instability, as well as non-motor-related symptoms such as olfactory deficiency, autonomic dysfunction, depression, cognitive impairment, and sleep disturbances. Like AD, PD is a proteinopathy characterized by the accumulation and aggregation of misfolded α-synuclein. Neuropathological features include intracellular inclusions containing α-synuclein called Lewy bodies and Lewy neurites, as well as the loss of dopaminergic neurons in the substantia nigra of the midbrain and other brain regions. The loss of dopaminergic neurons is not the only neuropathological change in PD, as it also results in increased microglial activation and increased astroglial and lymphocyte infiltration. Increased astroglial cells and increased number of dystrophic astrocytes in postmortem tissue from the brains of PD patients have also been reported. Multiple pieces of evidence suggest that inflammatory mediators induced from non-neuronal cells, including microglia, modulate the progression of neuronal cell death (or loss) in Parkinson's disease (PD).

[0004] Frailty and Parkinson's disease (PD) share a common characteristic: decreased activity in the PGC / AMPK / Sirt pathway. This pathway integrates the animal's primal stress response, influencing aging and increasing the risk of frailty. The PGC / AMPK / Sirt anti-stress pathway is activated by non-lethal levels of environmental cellular stress. Under stressful conditions, more energy is consumed to regulate homeostasis, so most environmental stressors ultimately decrease intracellular energy levels (AMP / ATP ratio). A decrease in the AMP / ATP ratio activates the AMPK enzyme, which phosphorylates various target molecules, resulting in an increase in the NAD / NADH ratio. An increase in the NAD / NADH ratio induces the activity of a class of proteins called sirtuins (Sirt). When AMPK and Sirt are activated together under stress, the PGC1 complex is activated. In its activated form, this complex activates the transcription of genes encoding proteins involved in the cell's anti-stress defense. This includes the formation of antioxidant enzymes, the formation of new mitochondria, and tighter closure of intercellular gap junctions. Interestingly, these high-stress conditions also induce uncoupling proteins (UCPs). These further reduce intracellular AMP / ATP levels, providing a positive feedback mechanism that allows cells to respond quickly to high-stress events through appropriate anti-stress responses.

[0005] Furthermore, PD and frailty share various common comorbidities and exhibit defects in the same biochemical pathway, suggesting that deregulation of this pathway plays a role in the pathogenesis of PD. The involvement of reduced activity of the PGC / AMPK / Sirt pathway in causing PD is further reinforced by experiments using resveratrol, which is known to activate this specific pathway, and in this case, resveratrol has the opposite effect on aging (i.e., frailty) and PD. The effects on comorbidities attributable to PD or frailty are similar, but these effects are the opposite of those seen with exercise or treatment using resveratrol. To further demonstrate that deregulation of the PGC / AMPK / Sirt pathway is involved in PD, the association with PD was studied using animal models in which the PGC / AMPK / Sirt pathway was reduced. Spontaneous hypertensive rats (SH rats) are a Wistar rat strain with spontaneous hypertension (Okamoto, 1963), and these animals show a significant reduction in the activity of the PGC / AMPK / Sirt pathway. These animals also exhibit all PD-related comorbidities. Spontaneously hypertensive rats suffer not only from elevated blood pressure but also from decreased bone mass, decreased muscle mass, shortened lifespan, altered microflora, and insulin resistance. All of these also occur in frailty, which is also associated with the PGC1 pathway. While these rats may develop PD, they do not exhibit the predicted rigidity or tremor, but in fact show a 50% reduction in Th-positive cells in their substantia nigra, an effect counteracted by exercise. This suggests that these rats suffer from a syndrome similar to early-stage PD. Taken together, these data indicate that a defective PGC / AMPK / Sirt pathway is involved in causing PD and increasing the risk of frailty.

[0006] In Parkinson's disease (PD), the enteric nervous system is affected well before motor symptoms appear, and the disease appears to spread from there. These neurons are close to the contents of the intestinal lumen, functionally separated from the intestine by a layer of intestinal cells. When this layer is damaged in PD patients, the enteric neurons are exposed to bacterial toxins such as LPS. This exposure to bacterial toxins is further amplified by the fact that Parkinson's disease-like patients suffer from dysbiosis of the microflora, resulting in more Gram-negative bacteria and thus more LPS production. If the primordial PGC / AMPK / Sirt anti-stress pathway is damaged as it is in PD, this may be sufficient to cause Parkinson's disease, as LPS further inhibits the PGC1 / AMPK / Sirt anti-stress pathway. In addition, studies conducted with Pink1 knockout mice have shown that asymptomatic Pink1 knockout mice became highly susceptible to PD symptoms after infection with Gram-negative (LPS-producing) bacteria. Mechanistically, these mice demonstrated a specific immune response against dopaminergic neurons under these conditions, and it was shown that Parkinson's disease-like symptoms induced by these bacteria could be alleviated by L-DOPA. Dependence on gut bacteria for the development of Parkinson's disease-like symptoms was also observed in alpha-synuclein overexpressing animals. Taken together, this evidence suggests that both genogenic and toxin-induced PD originate in the gut, and that gut bacteria strongly contribute to the development of PD.

[0007] In Parkinson's disease (PD), the PGC / AMPK / Sirt pathway is inhibited by genetic changes or exotoxins. Since this pathway controls intestinal barrier function, this increases exposure to bacterial toxins. Both of these changes activate the (pro-inflammatory) NFκB pathway in the body. The NFκB pathway is a common target for both bacterial toxins and the PGC / AMPK / Sirt pathway; bacterial toxins activate the NFκB pathway by activating TLR receptors, while the PGC / AMPK / Sirt pathway has its activity reduced. In PD, this leads to an imbalance in the activation of this pathway, resulting in pro-inflammatory changes in the body, which likely contribute to the neurological damage observed in PD. A crucial aspect of research into healing the neurodegeneration occurring in PD is finding targets that can be targeted with drugs. Directly targeting the NFκB or PGC / AMPK / Sirt pathway with drugs is difficult because both pathways are deeply embedded in the patient's overall physiology, making it challenging to select drugs with an effective and safe range of activity. Therefore, a natural approach that restores the natural balance of PD is preferable.

[0008] Considering the foregoing, in the art there is a need for the treatment of neurodegenerative diseases characterized by the gradual loss of neurons in different regions of the central nervous system (CNS), more preferably for the treatment of PD and / or frailty, and a need to find targets from which drugs can be administered for treatment that provide sustained and efficient treatment, and in PD and / or frailty, a need to restore the natural balance with respect to the NFκB or PGC / AMPK / Sirt pathway associated with the development of neurodegenerative diseases, more preferably PD and / or frailty. [Prior art documents] [Non-patent literature]

[0009] [Non-Patent Document 1] YAO, Chen, et al. "LRRK2-mediated neurodegeneration and dysfunction of dopaminergic neurons in a Caenorhabditis elegans model of Parkinson's disease", Neurobiology of disease, 2010, 40.1: 73~81 [Non-Patent Document 2] Leveille et al, 2020, Molecular Metabolism 34, 72~84 [Non-Patent Document 3] Yuan Luo et al. in Methods of Behavior Analysis in Neuroscience. 2nd edition, Chapter 16, "Caenorhabditis elegans Model for Initial Screening and Mechanistic Evaluation of Potential New Drugs for Aging and Alzheimer's Disease" [Non-Patent Document 4] Bergmeyer HU (1974) Methods of Enzymatic Analysis, 2nd edition, p496, Academic Press, New York. [Non-Patent Document 5] Waqar Ahmand et al. 2017, Molecular Neurobiology, 54, 5427~5439 [Overview of the project] [Problems that the invention aims to solve]

[0010] One objective of the present invention is, among other objectives, to address the aforementioned needs in the art. The objectives of the present invention are, among other objectives, satisfied by the invention outlined in the appended claims. [Means for solving the problem]

[0011] Specifically, the above-mentioned objectives, among other objectives, are, in particular, alkaline phosphatase for use in the treatment or prevention of a mammal suffering from or at risk of a neurodegenerative disorder caused by a decrease in peroxisome proliferator-activated receptor gamma coactivator 1 (PGC1) activity compared to a healthy individual, according to a first aspect of the present invention, wherein the treatment comprises administering to the mammal a therapeutically effective amount of alkaline phosphatase that inhibits neurodegeneration or neurodegradation by preventing a decrease in PGC1 activity. Administration of alkaline phosphatase (AP) results in a reduction or prevention of neurodegeneration or neurodegradation, thereby reducing or preventing neurodegenerative disorders, including Parkinson's disease, Alzheimer's disease, and frailty.

[0012] Experiments demonstrate that treatment with alkaline phosphatase protects genetically modified Caenorhabditis elegans (Parkinson's disease-like nematodes) from dopaminergic neurodegeneration. A G2019S LRRK2 nematode model was used, as described in YAO, Chen, et al., "LRRK2-mediated neurodegeneration and dysfunction of dopaminergic neurons in a Caenorhabditis elegans model of Parkinson's disease," Neurobiology of Disease, 2010, 40.1: 73-81. In this animal model, the native LRRK gene of the nematode was removed and replaced with a human G2019S LRRK2 variant that causes Parkinson's disease in humans. In addition, the inventors demonstrate that alkaline phosphatase extends the lifespan of these Parkinson's disease-like nematodes. Neuronal susceptibility to Parkinson's disease-causing mutations is evolutionarily conserved in the PGC / AMPK / Sirt pathway, and the effect of alkaline phosphatase on lifespan is evolutionarily conserved because the anti-stress PGC / AMPK / Sirt pathway involved is an evolutionarily conserved pathway in mammals. Its effects include the following targets: AMP / ATP ratio, AMPK, NAD / NADH ratio, SIRT-1, PGC1, and the uncoupling protein UCP. All of these targets together regulate the anti-stress response of a common cell. PGC1 refers to a family of transcriptional coactivators that coordinately regulate metabolic pathways and biological processes in a tissue-specific manner. The PGC1 family consists of various family members, including at least PGC1 alpha and PGC1 beta, and further subfamily members, of which PGC1 alpha 4 may be an example. In various organs, different family members and subtypes are often expressed in a coordinated manner to enable precise metabolic control of various organs, as already described by Leveille et al, 2020, Molecular Metabolism 34, 72-84.

[0013] Furthermore, a memory test using the genetically modified nematode CL2355 tested the therapeutic effect of AP on Alzheimer's disease (AD). This experiment utilized a genetically modified nematode strain (CL2355), as already described by Yuan Luo et al. in Methods of Behavior Analysis in Neuroscience, 2nd edition, Chapter 16, "Caenorhabditis elegans Model for Initial Screening and Mechanistic Evaluation of Potential New Drugs for Aging and Alzheimer's Disease." CL2355 expresses human A-beta (Aβ) in a temperature-dependent manner in all of its neurons; that is, A-beta expression only begins when the animal's growth temperature shifts from 16°C to 25°C. Due to the expression of human A-beta, these animals develop memory impairment corresponding to the progression of AD disease in humans, which is also associated with A-beta expression in neurons and memory loss. The results show that treatment of nematodes with 111, 333, and 1000 IU / drop of AP statistically significantly increased memory in CL2355 animals in a dose-dependent manner compared to buffer-treated controls. Therefore, treatment with AP counteracts memory loss in a dose-specific manner.

[0014] The cellular anti-stress response mediated by PGC1 is shown in Figure 4. With small ATP deficiencies (-) or AMP excesses (+), AMPK is activated, resulting in increased mitochondrial ATP production. In more severe ATP deficiencies, CPT1 is activated by AMPK, burning fatty acids released from stored fats. If ATP requirements remain high and CPT1 remains activated, intracellular NAD / NADH levels increase. This parameter signifies an increased oxidative (stress) state, which activates Sirt1. Sirt1 deacetylates PGC1, AMPK phosphorylates PGC1, and PRMT1 methylates PGC1, thereby activating PGC1 to recruit the nuclear factor PPAR delta. Activated PGC1 and PPAR delta bind to the promoter regions of genes involved in protecting cells from metabolic and redox stress, enhancing their transcription and providing a positive feedback loop for AMP / ATP and further anti-stress responses. This anti-stress response can be activated by alkaline phosphatase by increasing the AMP / ATP ratio (activating AMPK) and by detoxifying LPS (inhibiting AMPK activity). This indirectly activates PGC1, which plays a crucial role in causing Parkinson's disease.

[0015] Peroxisome proliferator-activated receptor gamma coactivator 1 (PGC1) acts as a stress sensor in cancer cells and can be activated by nutritional deficiencies, exercise, and oxidative damage. PGC1 influences mitochondrial respiration, reactive oxygen species defense systems, and fatty acid metabolism by interacting with specific transcription factors, affecting the regulation of both carbohydrate and lipid metabolism. PGC1 is one of the nuclear factors activated by AMPK activation and (like AMPK) plays a crucial role in anti-stress biochemistry. PGC1 acts as an essential link connecting metabolic regulation, redox control, and inflammatory pathways, making it an interesting therapeutic target for neurodegenerative diseases, such as Parkinson's disease (PD). AMPK is an indirect target for the treatment of PD because it affects the activation of PGC1 in cells. Parkinson's disease and other neurodegenerative diseases involve dysregulation of the PGC1 nuclear factor. Currently, there are no direct activators of PGC1. AP is used to activate PGC1 by activating and regulating AMPK; i.e., AP-mediated PGC1 activation is indirect. This allows the body to naturally protect itself from direct overactivation of PGC1, for example, by modulating AMP formation through adenosine kinase and other biochemical pathways.

[0016] The mechanism of action of exercise mimics (which include biological factors that can improve human and animal endurance without the need for training) has shown that the same biochemical pathway is triggered by these exercise mimics, just as it is by exercise itself, and that this same pathway is also important for achieving extended healthy lifespan. Activation of the AMPK / Sirt / PGC1 / PPAR complex was found to be crucial in this regard. By comparing the biochemistry of lifespan with that of Parkinson's disease, it became clear that PD (like frailty) is likely caused by a decrease in PGC1 activity. Alkaline phosphatase protects against neurodegeneration in human Parkinson's disease by activating the PGC1 anti-stress pathway, and the protective effect of alkaline phosphatase against dopaminergic neurodegeneration is evolutionarily conserved. This is further supported by the fact that differential gene expression in PD is largely regulated by PGC1, PGC1 expression is lower in PD patients, aging (which enhances PD and frailty-related disorders) reduces PGC1 expression, exercise (which inhibits PD neurodegeneration) activates PGC1, and a decrease in PGC1 in animals leads to dopaminergic neurodegeneration. For example, neurodegeneration in PD is caused by PGC1 inactivation, resulting in a defect in the cellular anti-stress response. AP activates the cellular anti-stress response, thereby reducing neurodegeneration. [Modes for carrying out the invention]

[0017] In a preferred embodiment, the present invention relates to an alkaline phosphatase for use in the treatment or prevention of a mammal suffering from or at risk of a neurodegenerative disorder, wherein the neurodegenerative disorder is selected from the group consisting of Alzheimer's disease (AD), Parkinson's disease (PD), frailty-related disorders, amyotrophic lateral sclerosis (ALS), multiple sclerosis (MS), and neurodegeneration as a result of stroke, preferably Parkinson's disease, Alkinson's disease, or frailty-related disorders. Examples of neurodegenerative diseases targeted by AP include AD, ALS, neurodegeneration as a result of stroke, and myalgic encephalomyelitis. Pathological features of AD in the brain include extracellular amyloid plaques containing aggregated and cleaved products of amyloid precursor protein (APP) and intracellular neurofibrillary tangles (NFTs), resulting from a hyperphosphorylated form of microtubule-associated protein tau. Evidence of the inflammatory response in AD includes changes in microglial morphology from branched (quiescent) to amoeboid (active), and astrogliosis surrounding senile plaques (manifested by an increase in the number, size, and motility of astrocytes). While the precise pathophysiological mechanisms underlying neurodegeneration in ALS remain unclear, common pathological features include the presence of ubiquitin immunoreactive cytoplasmic inclusions in degenerated neurons, followed by a strong inflammatory response. Marked neuroinflammation can readily be observed in pathological sites of the CNS and spinal cord. Typically, inflammation in ALS is characterized by gliosis and the accumulation of numerous activated microglia and astrocytes. MS is an autoimmune disease characterized by inflammation, demyelination, and axonal degeneration in the CNS, more specifically by the infiltration of lymphocytes and antibody-producing plasma cells into the perivascular regions of the brain and spinal cord white matter, an increase in microglia and astrocytes, and demyelination. Frailty is a part of the aging process in which the body gradually loses its natural reserves, making it vulnerable to dramatic and rapid changes in health, including neurodegeneration, which can be triggered by seemingly minor events such as mild infections or changes in medication or environment. Frailty is generally characterized by problems such as muscle weakness and fatigue.

[0018] In a preferred embodiment, the present invention relates to an alkaline phosphatase for use in the treatment or prevention of a mammal, preferably a human, suffering from or at risk of neurodegenerative disorders, wherein preventing a decrease in PGC1 activity involves increasing the activation of adenosine monophosphate-activated protein kinase (AMPK) by the alkaline phosphatase, thereby promoting an anti-stress response. The alkaline phosphatase activates AMPK by inducing AMP production when stressed adjacent cells release ATP into the cellular environment. This ATP is metabolized to adenosine by the extracellular alkaline phosphatase. Consequently, target cells take up the adenosine formed by the alkaline phosphatase, phosphorylate it to AMP intracellularly, thereby activating AMPK. Alternatively, adenosine formed from ATP by the alkaline phosphatase activates purinergic surface receptors on target cells, thereby also activating AMPK. Furthermore, AP also detoxifies LPS, a pro-inflammatory molecule that inhibits AMPK activity in target cells.

[0019] In a preferred embodiment, the present invention relates to an alkaline phosphatase for use in the treatment or prevention of neurodegenerative disorders in mammals suffering from or at risk thereof, wherein the therapeutically effective dose is 6 to 1350 U / day / kg, preferably 25 to 750 U / day / kg, more preferably 50 to 500 U / day / kg. AP activity is defined in (glycine) units / ml, as disclosed in Bergmeyer HU (1974) Methods of Enzymatic Analysis, 2nd edition, p496, Academic Press, New York. One unit is the amount of enzyme capable of converting 1 μM of para-nitrophenol phosphate (PNPP) per minute at 25°C and pH 9.6 (glycine buffer). Experiments using AP have been conducted in nematodes, and these experiments have shown that AP significantly extends lifespan at 1000 U compared to an untreated group. The mean lifetimes of untreated and treated nematodes with 1000 U of AP were found to be 14.93 + 0.52 days and 17.37 + 0.92 days, respectively. When the AP dose was reduced to 200 U, the effect of AP was still observed, although it was not as pronounced. Based on these results, it is estimated that the predicted therapeutic effective dose of AP for mammals, more specifically for humans, is between 500 and 100,000 U per person per day, preferably between 5,000 and 30,000 U (average body weight approximately 75 kg), or the therapeutic effective dose of AP for humans is between 6.7 and 1333.3 U / day / kg, preferably between 66.7 and 400 U / day / kg.

[0020] In a preferred embodiment, the present invention relates to an alkaline phosphatase for use in the treatment or prevention of mammals suffering from or at risk of neurodegenerative disorders, wherein the alkaline phosphatase is a tissue-specific ectophosphatase selected from the group consisting of intestinal AP (IAP), placental ALP (PALP), and hepatic AP (LAP), preferably IAP or PALP. PD can be treated using intestinal alkaline phosphatase (IAP). Tissue-nonspecific ectophosphatases may also be suitable for use in the treatment of the present invention. PD begins with decreased activity of the PGC / AMPK / Sirt pathway. This decreased activity may have a genetic cause, but toxins, such as LPS, also decrease the activity of this pathway. SH rats, which exhibit decreased PGC / AMPK / Sirt pathway activity, also show a potent decrease in intestinal alkaline phosphatase levels. Activators of the PGC / AMPK / Sirt pathway, including oleic acid and curcumin, increase IAP expression in the gut. IAP has a significant effect on the intestinal microflora, and decreased alkaline phosphatase activity leads to dysbiosis of the microflora in PD. As observed in PD, decreased activity of the PGC / AMPK / Sirt pathway leads to downregulation of tight junctions. In the gut, this increases exposure to bacterial toxins, including LPS, resulting in immune activation through exposure of TLR receptors in the gut immune system. In the brain, this increases the permeability of the blood-brain barrier to inflammatory toxins and circulating immune cells. Increased exposure of neural tissue to bacterial toxins increases alpha-synuclein expression. This expression promotes LPS-induced blood-brain barrier leakage and activation of the immune system targeting the brain in PD.

[0021] According to a preferred embodiment, the present invention relates to an alkaline phosphatase for use in the treatment or prevention of a mammal suffering from or at risk of a neurodegenerative disorder, wherein the treatment comprises intravenous, parenteral or oral administration, preferably oral administration. For oral administration, IAP is preferred, and for parenteral use, PALP is more preferred. Oral alkaline phosphatase can be a treatment for PD because the treatment using it promotes the growth of commensal bacteria in the intestine, promotes the closure of the intestinal barrier, and detoxifies various bacterial toxins including LPS and ATP. As a result, AP normalizes the intestinal flora, closes the leaky gap junctions in the intestine, reduces neuroinflammation induced by these changes and by activating the efferent nerve terminals of the vagus nerve, thereby preventing the development of PD. The present invention is supported by the further observation that orally administered IAP can be used for the treatment of PD. Studies have shown that the adaptive immune response leading to PD is controlled by Th17 cells. Th17 cells mature in the intestine only in the presence of a suitable pro-inflammatory intestinal microflora, and this maturation changes in response to luminal ATP in the intestine. Since IAP rapidly dephosphorylates luminal ATP, it will also inhibit Th17 cell activation. Thus, orally administered IAP plays an important role in steering the adaptive immune response and can prevent the development of PD.

[0022] Furthermore, as shown above, the PGC / AMPK / Sirt pathway is affected in PD. This pathway is also involved in lifespan extension. The results of the present inventors show that administration of IAP to nematodes significantly extended their lifespan, suggesting that IAP can activate the systemic PGC / AMPK / Sirt pathway by acting only in the intestine. Oral AP improves the symptoms of metabolic syndrome, which is a known important risk factor for PD. The gut microbiota of PD patients modified by probiotics has been shown to have a positive effect on PD symptoms, as well as insulin sensitivity and plasma triglycerides. However, probiotics often have limited effects on gut microbiota. The effect of treatment with AP is preferred because it is persistent and more natural, with a high likelihood of restoring normal microbiota, while the effect of treatment with probiotics varies depending on the exact strain or mixture of strains of bacteria administered.

[0023] According to a preferred embodiment, the present invention relates to an alkaline phosphatase for use in the treatment or prevention of a mammal suffering from or at risk of a neurodegenerative disorder, which is a recombinant alkaline phosphatase, preferably a recombinant mammalian alkaline phosphatase, more preferably a human recombinant alkaline phosphatase. Preferably, the phosphatase used in the composition of the present invention is compatible with a therapeutic intervention that is expected to assist in the treatment of a human using the composition of the present invention, for example, comprising recombinant human alkaline phosphatase. However, other combinations can also be used, for example, using the treatment of a human using the composition of the present invention comprising a non-human native or non-human recombinant alternative alkaline phosphatase, such as alkaline phosphatase derived from bovine or porcine intestine.

[0024] In a preferred embodiment, the present invention relates to an alkaline phosphatase for use in the treatment or prevention of mammals suffering from or at risk of neurodegenerative disorders, wherein the alkaline phosphatase is one or more selected from the group consisting of a biologically active fragment or derivative of an alkaline phosphatase, a synthetic alkaline phosphatase derivative, and a chemical-pharmaceutically small molecule that exhibits functional alkaline phosphatase activity, preferably a biologically active fragment or derivative of an alkaline phosphatase. The biologically active fragment or derivative of an alkaline phosphatase enables recovery and protects the functional properties and integrity of the blood-brain barrier, the intracerebral lymphoid system, and the functional neurosupport properties of the microglia and astroglial systems of the central nervous system.

[0025] In a preferred embodiment, the present invention relates to an alkaline phosphatase for use in the treatment or prevention of a mammal suffering from or at risk of a neurodegenerative disorder, wherein the treatment or prevention includes delaying the onset of the neurodegenerative disorder, or attenuating or preventing its progression. Acute-onset neurodegenerative disorders (e.g., stroke) and chronic neurodegenerative disorders (e.g., AD, PD, MS) involve persistent inflammatory responses. Each of these disorders is distinguished by disease-specific mechanisms for inducing the inflammatory response. The distinct pathways for inducing inflammation and the specific anatomical locations in which these processes occur are likely determinants of the specific pathological features of each neurodegenerative disease. However, it is noteworthy that, once inflammation is induced, there appears to be a significant convergence of mechanisms leading to amplification of the inflammatory response, neurotoxicity, and neuronal cell death. Activation of innate immune cells in the CNS, e.g., microglia and astrocytes, is one of the common components of neuroinflammation. In affected central nervous system systems (CNS), the interaction between damaged neurons and dysregulated, hyperactivated microglia creates a severe self-propagating cycle that leads to uncontrolled, prolonged inflammation, and this cycle drives the chronic progression of neurodegenerative diseases.

[0026] In a preferred embodiment, the present invention relates to an alkaline phosphatase for use in the treatment or prevention of a mammal suffering from or at risk of a neurodegenerative disorder, wherein the treatment comprises attenuating the inflammatory response of the mammal suffering from the neurodegenerative disorder. In many inflammatory conditions, AP has been shown to safely and effectively target inflammatory mechanisms, as well as those contributing to the pathogenesis of various neurodegenerative disorders. Since neurodegenerative disorders are chronic diseases, their prevention and treatment are likely to require long-term treatment and thus impose requirements for high safety levels. Clinical studies conducted with AP have not shown any signs of adverse activity in patients. Furthermore, in repeated dose-toxicity studies in various animal species immune to this protein, animals have shown tolerance to high-dose intravenous injection of AP once daily. Therefore, the inventors predict that AP can be safely applied in patients with neurodegenerative disorders.

[0027] In a preferred embodiment, the present invention relates to an alkaline phosphatase for use in the treatment or prophylaxis of mammals suffering from or at risk of neurodegenerative disorders, wherein the treatment or prophylaxis involves promoting the activation of an anti-inflammatory cytokine selected from the group consisting of IL-1, IL-4, IL-6, IL-10, IL-11, and IL-13, preferably IL-11. Non-neuronal microglia cells, as with mononuclear leukocytes, exert immunomodulatory functions similar to those of macrophages in circulation, and are activated and inactivated by pro-inflammatory and anti-inflammatory factors. LPS and ischemic inflammatory conditions are resolved by AP activity. Preliminary in vitro studies in mouse BV2 microglia cells demonstrate that AP distorts the activation profile of ATP-stimulated microglia in favor of the M2 anti-inflammatory phenotype, as inferred from the selective increase in the expression of the anti-inflammatory M2 marker cytokine IL-10 (results not shown). AP also reduces the M1 pro-inflammatory activation of LPS-stimulated microglia by decreasing the mRNA expression levels of the pro-inflammatory M1 marker cytokines TNF-α, IL-6, and IL-Iβ.

[0028] In a further embodiment, the present invention relates to a method for inhibiting neurodegradation by preventing a decrease in PGC1 activity in a mammal, the method comprising the step of administering a therapeutically effective amount of the above-defined alkaline phosphatase to the mammal.

[0029] In another embodiment, the present invention relates to the use of alkaline phosphatases disclosed herein for the preparation of pharmaceuticals for the prevention of neurodegenerative disorders in mammals at risk due to reduced PGC1 activity compared to healthy individuals.

[0030] The present invention is further described in detail by the following embodiments and figures. [Brief explanation of the drawing]

[0031] [Figure 1]The survival curve of the gene-transfected LRRK2-G2019S nematode is shown. The lifespan of nematode mutants carrying the human G2019S LRRK2 mutation can be extended by increasing the amount of externally added alkaline phosphatase (200 U(A) and 1000 U(B) AP). Based on this observation, since the effect of alkaline phosphatase on lifespan is evolutionarily conserved, it is predicted that alkaline phosphatase will extend the lifespan of worms and mammals under standard survival conditions (similar pathways, similar mechanisms). [Figure 2] The percentage of dopaminergic (DA) neurons that survived (intact DA neurons %) in adult LRRK2-G2019S nematodes grown in the absence and presence of 1000U(A) or 200U(B) of AP is shown over time at days 1, 4, 7, and 12. [Figure 3] Fluorescence microscopy images of GFP-tagged DA neurons in adult LRRK2-G2019S nematodes grown in the absence and presence of 200U of AP are shown. [Figure 4]This diagram illustrates the regulation of the cellular anti-stress response by PGC1. The regulators within the ellipse are involved in the cellular transcriptional stress response, with PGC1 at its very center. With small ATP deficiencies (-) or AMP excesses (+), AMPK is activated, resulting in increased mitochondrial ATP production. In more severe ATP deficiencies, CPT1 is activated by AMPK, burning fatty acids released from stored fats. If ATP requirements remain high and CPT1 remains activated, intracellular NAD / NADH levels will increase. This parameter signifies an increased oxidative (stress) state, which activates Sirt1. Sirt1 deacetylates PGC1, AMPK phosphorylates PGC1, and PRMT1 methylates PGC1, thereby activating PGC1 to recruit the nuclear factor PPAR delta. Activated PGC1 and PPAR delta bind to the promoter regions of genes involved in protecting cells from metabolic and redox stress, enhancing their transcription and providing a positive feedback loop for AMP / ATP and further anti-stress responses. This anti-stress response can be activated by alkaline phosphatase by increasing the AMP / ATP ratio (activating AMPK) and by detoxifying LPS (inhibiting AMPK activity). This indirectly activates PGC1, which plays a crucial role in causing Parkinson's disease. [Figure 5]The results of memory tests in trained nematodes CL2122 (control group) and CL2355 (Alzheimer's disease model group) after treatment with different doses of AP or metformin (positive control) are shown. Memory is expressed as a chemotactic index, taking into account the respective treatments in CL2122 and CL2355 animals. The chemotactic index is calculated as the number of animals in the ethanol spot minus the number of animals in the non-spotted area, divided by the total number of animals, as determined in the memory test (see Example 2 for details). Memory in CL2355, which expresses A-beta, i.e., human A-beta resulting in severe memory loss, was close to zero. CL2122 does not express A-beta and serves as the control group. AP shows a dose-response that takes into account the improvement in memory in nematode CL2355. Treatment with 111, 333, and 1000 IU / drop of AP infectant statistically significantly increased memory in CL2355 animals in a dose-dependent manner compared to buffer-treated controls. Metformin-treated animals also showed a statistically significant increase in memory in this study. [Examples]

[0032] (Example 1) In vivo effects of AP on the lifespan and neurodegeneration of the genetically modified LRRK2-G2019S nematode. Here, we investigated the effects of AP on the protection of dopaminergic neurons in an animal model of Parkinson's disease (PD). In this study, we assessed the effects of AP on lifespan and neurodegeneration using a transgenic LRRK2-G2019S nematode. We used the G2019S LRRK2 nematode model as described in YAO, Chen, et al., "LRRK2-mediated neurodegeneration and dysfunction of dopaminergic neurons in a Caenorhabditis elegans model of Parkinson's disease," Neurobiology of Disease, 2010, 40.1: 73-81. In this animal model, the native LRRK gene of the nematode was removed and replaced with the human G2019S LRRK2 variant, which causes Parkinson's disease in humans. The nematode contains the AAK, Sir2.1, and MDT15 / NHR49 complex, which are orthologs of AMPK, Sirt1, and PGC1, respectively. AAK and Sir2.1 (and naturally AMPK and Sirt1 as well) are enzymes that alter the activity of other proteins. This ultimately activates a protein complex in the cell nucleus that triggers DNA transcription of anti-stress genes. We used this genetically modified nematode to assess the effect of AP on the survival of dopaminergic neurons.

[0033] The following experiments were conducted. A lifespan experiment was performed to determine the effectiveness of AP in extending the lifespan of genetically modified LRRK2-G2019S nematodes grown in live fungal colonies. Furthermore, a neurodegeneration experiment was conducted to test the effectiveness of AP in protecting dopaminergic neurons from degeneration during aging and frailty-related symptoms in genetically modified nematodes grown in live fungal colonies. Subsequently, the above two experiments were repeated to confirm the effects of AP on both lifespan extension and neuroprotection in genetically modified LRRK2-G2019S nematodes. In addition, microscopic images of dopaminergic neurons were taken during the neurodegeneration experiment.

[0034] Nematode growth media and agar plates were prepared as follows. An agar solution was prepared by dissolving 3 g of NaCl, 17 g of agar, and 2.5 g of peptone in 975 ml of redistilled water using a stirring rod and stirring plate. The agar solution was autoclaved at 121°C for 30 minutes using a liquid cycle with 500 ml of distilled water via a dispenser tube. The agar solution was cooled to 75°C and simultaneously stirred on a stirring plate. Using sterilization techniques, 1 ml of 4 ml of 1 M MgSO4, 1 ml of 2 ml of 1 M CaCl, 1 ml of 5 mg / ml cholesterol, and 1 ml of 4 ml of 1 M KPO4 were added to the agar solution. For 35 mm plates, 1 ml of FUdR (75 μM stock solution) was also added to the agar solution. For 35 mm plates, 4 ml of FUdR and agar solution were dispensed into each plate to provide FUdR+ plates. For 60 mm plates, 8 ml of agar solution was dispensed into each plate. The plates were stored at room temperature for two days, then covered with a plastic tray sterilized with 70% EtOH to allow the agar to solidify completely.

[0035] E. coli OP50 solution was prepared by collecting E. coli OP50 from a pre-inoculated agar plate by scraping the pipette tip across the bacterial flora using a sterile pipette tip. The pipette tip was incubated overnight in 1 liter of LB broth at 37°C. The E. coli solution was stored in a 4°C cooler until inoculation onto the agar plate. Using the E. coli OP50 solution, 50 μl was pipetteed onto a 35 mm plate, and 150 μl onto a 60 mm plate. The inoculated plates were covered with a sterilized plastic tray with 70% EtOH and stored at room temperature until the bacterial flora developed. The inoculated plates containing the E. coli flora were stored in a 4°C cooler until use.

[0036] Alkaline phosphatase (AP) solution was prepared by dissolving AP in a buffer solution consisting of 20 mM Tris (pH 7.8), 5 mM MgCl2, and 0.1 mM ZnCl2 (AP buffer), resulting in a concentration of 50,000 units per ml. The solution was divided into fixed volumes of 100 μl and stored at 4°C.

[0037] Before initiating any assay, the age of the transgenic LRRK2-G2019S nematodes was age-synchronized to induce development from eggs after alkaline bleaching, thereby matching the age of the nematode samples. To do this, 10 μl of bleaching solution (25 μl of 5M NaOH, 100 μl of 8% bleach, and 375 μl of redistilled water) was pipetteed onto a 60 mm plate away from the microbial community. Fifteen L4 stage nematodes were selected from the cultured population and placed in the bleaching solution on the 60 mm plate using a sterile platinum wire. Further bleaching solution was added as evaporated solution from the plate until all eggs were released. This procedure was repeated in a second spot on the same 60 mm plate. The eggs were allowed to hatch and grow for two days, and 30 hatched nematodes were transferred to each 35 mm FUdR+ plate at the start of each assay.

[0038] 35 mm FUdR+ plates were divided into three groups: one treated with 4 μl AP (200 U) solution, one with 20 μl AP (1000 U) solution, and a control group. Treatment plates were processed by pipetting the respective volumes of AP solution onto the microbial colony of the plate and immediately transferring the nematodes. The plates were tilted to spread the solution across the colony and ensure that the solution coated the entire colony. Separate control groups were maintained alongside each treatment group, scored, and transferred on the same day. All plates were dried for 15 minutes after solution application before transferring the nematodes.

[0039] Lifetime assay Each experiment consisted of three treatment plates containing either 20 μl or 4 μl of AP, and three control plates. Nematodes were scored and moved daily until oviposition ceased. Nematodes were scored every 1–3 days, and moved every 2–4 days after oviposition ceased. Nematodes were scored as dead if they did not move in response to light contact using a sterile platinum wire, or if they were alive but could not be moved (e.g., trapped under agar), or appeared to have died due to an abnormal cause, such as hatching inside the nematode's body. This continued until no nematodes remained.

[0040] In this study, two independent groups of transgenic LRRK2-G2019S nematodes were treated with either 200 U or 1000 U of AP. Based on the results, survival curves were obtained for transgenic LRRK2-G2019S nematodes grown in the absence of 1000 U of AP (Figure 1A) and 200 U of AP (Figure IB). This confirmed that 1000 U of AP significantly extended the lifespan. The mean lifespans of untreated nematodes and those treated with 1000 U of AP were found to be 14.93 + 0.52 days and 17.37 + 0.92 days, respectively. When the AP dose was lower than 200 U, the effect of AP was significantly reduced. The mean lifespans of untreated nematodes and those treated with 200 U of AP were found to be 14.34 + 0.50 days and 14.64 + 0.38 days, respectively.

[0041] AP activity is defined in (glycine) units / ml, as disclosed in Bergmeyer HU (1974) Methods of Enzymatic Analysis, 2nd edition, p496, Academic Press, New York. From these results, it is estimated that the predicted therapeutic effective dose of AP for mammals, more specifically humans, is between 500 and 100,000 U per person per day, preferably between 5,000 and 30,000 (average body weight approximately 75 kg), or the therapeutic effective dose of AP for humans is between 6.7 and 1333.3 U / day / kg, preferably between 66.7 and 400 U / day / kg.

[0042] Neurodegeneration assay Each experiment was initiated using four treatment plates containing either 20 μl or 4 μl of AP, and four control plates. The viable nematodes were moved daily until oviposition ceased, and then every 2–4 days thereafter. The number of viable dopaminergic (DA) neurons in 10 representative nematodes from each group was counted using a fluorescence microscope on day 1 or 2 of the experiment, and then every 3–4 days from four different days until sample collection. If fewer than 10 nematodes were viable on the final day, the viable nematodes were scored, and the remaining 10 were marked as dead and containing no viable neurons. To score the neurons, adhesive plastic binding-reinforced rings were attached to the surface of the microscope glass slides. Two μl of 1x mounting solution (10x stock solution: 10 mg of 1% tricaine and 1 mg of 0.1% tetramizole dissolved in water) and four μl of redistilled water were pipetteed into the center of the adhesive plastic ring on a glass slide. Ten nematodes from the treatment group samples were transferred to the solution on the slide. A coverslip was placed on the slide, and the nematodes were immobilized with the mounting solution. The coverslip was then attached to the slide using superglue. If the nematodes were not immobilized with the mounting solution within five minutes, an additional 2 μl of mounting solution was added. Further mounting solution was added every five minutes until the nematodes were successfully immobilized.

[0043] GFP-tagged dopaminergic neurons from living nematodes were observed under a fluorescence microscope. The number of healthy DA neurons was counted using the fluorescence microscope. The number of deficient and unhealthy neurons was also recorded. Neurons were considered unhealthy if their cell bodies appeared shrunken, or if their axons were damaged or beaded. This was repeated for control samples. Dopaminergic neurons were scored for signs of degeneration resulting from cell body deficity and shrunkenness or neurite damage. The number of intact neurons was plotted against the age of the nematodes in each group (Figure 2).

[0044] Two independent groups of transgenic LRRK2-G2019S nematodes treated with either 200U (Figure 2A) or 1000U (Figure 2B) of AP demonstrated that AP protects dopaminergic neurons from age-dependent degeneration in both 200U and 1000U doses. AP significantly enhanced the survival of dopaminergic neurons measured at adult days 4 (p<0.001) and 7 (p<0.001) with 200U of AP, and at adult day 7 (p<0.001) with 1000U of AP. At 7 days postna (equivalent to approximately 50 years of age in humans), approximately 50% of dopaminergic neurons were damaged in these mutant nematodes, which corresponds to the early onset of Parkinson's disease in nematodes and is comparable to the situation observed in Parkinson's disease patients.

[0045] Consistent with the DA neuron counts described above, AP attenuated the age-dependent decline in GFP signaling in DA neurons of LRRK2-G2019S nematodes, indicating enhanced DA neuron survival after AP treatment. Representative images of DA neurons in LRRK2-G2019S nematodes treated with 200 U of AP on adult days 1, 4, and 7 are shown in Figure 3.

[0046] Discussion The experiment showed that the lifespan of transgenic nematodes was extended by the addition of 1000 U of AP, supporting the finding that AP promotes healthy aging and reduces frailty-related disorders in a nematode model of Parkinson's disease (PD). These data support the finding that AP can be used to activate anti-stress biochemistry, thereby regulating lifespan and anti-stress pathways, and thus neuroprotection should occur. Furthermore, the experiment showed that AP protects dopaminergic neurons from age-dependent degeneration in transgenic LRRK2-G2019S nematodes. This in vivo effect was statistically significant at both 200 U / plate and 1000 U / plate. The protective effect of AP on dopaminergic neurons in the same neurodegeneration model is comparable to or better than the protective effects of drugs (GW5074, sorafenib, and AdoCbl) that were previously reported to be effective in this PD nematode model. These results confirmed the beneficial effects of AP on lifespan and neurodegeneration in the transgenic LRRK2-G2019S nematode, a model of Parkinson's disease. 1000 U of AP was shown to extend lifespan, while 200 U and 1000 U of AP were shown to protect against dopaminergic neurodegeneration occurring in the transgenic LRRK2-G2019S nematode. Higher doses of AP appear to be required for lifespan extension, while neuroprotective effects can be detected at relatively lower doses.

[0047] (Example 2) In vivo effect of AP on Alzheimer's disease memory tests in genetically modified CL2355 nematodes In further experiments, the therapeutic effect of AP on Alzheimer's disease (AD) was tested. This experiment utilized a genetically modified nematode strain (CL2355), as already described by Yuan Luo et al. in Methods of Behavior Analysis in Neuroscience, 2nd edition, Chapter 16, "Caenorhabditis elegans Model for Initial Screening and Mechanistic Evaluation of Potential New Drugs for Aging and Alzheimer's Disease." CL2355 expresses human A-beta (Aβ) in a temperature-dependent manner in all of its neurons; that is, A-beta expression only begins when the animals' growth temperature shifts from 16°C to 25°C. Due to the expression of human A-beta, these animals develop memory impairment equivalent to the progression of AD disease in humans, which is also associated with A-beta expression in neurons and memory loss. The nematode strain CL2122 was used as a control strain. This strain shares the same genetic background as CL2355, but it completely fails to express A-beta when the temperature shifts.

[0048] This study investigates the effect of alkaline phosphatase (AP) on A-beta-induced memory loss in CL2355 nematodes. Metformin, an AMPK activator known to enhance memory in A-beta-expressing CL2355 animals, was used as a positive control, as disclosed by Waqar Ahmand et al. 2017, Molecular Neurobiology, 54, 5427-5439. After treatment with either the test compound, AP, or metformin, the animals were starved for 1 hour. Subsequently, the animals were placed on a standard NGM agar plate for 1 hour in the presence or absence of butanone (training condition) or in the absence of butanone (untrained) to train them. After this time, the animals were starved again for 1 hour and prepared for testing of memory retention.

[0049] Preparation of exposure plates for test compounds NGM agar plates, which would be used to expose animals to the test compound, were prepared four days before the memory test and stored in the dark at 4°C. To prepare the test compound-containing plates, 10 drops of 3x concentrated standard OP50 bacterial culture (50 µl) were pipetteed into each 10 cm diameter NGM plate. After these droplets dried, the test compound was added.

[0050] To administer a negative control, AP was dissolved in 20 µl of buffer. The buffer consisted of 20 mM Tris buffer, 5 mM MgCl23, and 0.1 mM ZnCl2. AP was tested at three different dose levels: 2.2, 6.6, and 20 µl of AP solution (50,000 IU / ml) corresponding to 111, 333, and 1000 IU of AP. Metformin was administered in a solution of 2.5 µl of 1.5 mM aqueous solution.

[0051] Exposure of nematodes to test compounds Approximately 1000 animals synchronized at stage L4 (grown at 16°C) were exposed to the test compound for 24 hours at 16°C by pipetting the animals onto plates containing the test compound, as described above. After these 24 hours, the animals were placed on unused plates and the test compound and incubated at 25°C for a further 24 hours. Memory retention was then determined by conducting a memory retention test as described below.

[0052] Memory retention test, chemotactic index To prepare the NGM agar plate for the memory test, pipette two drops of 1µl of 1M sodium azide onto a 10cm diameter agar plate, placing them approximately 6cm apart in the center of the plate. The presence of azide will paralyze animals that approach these spots. Then, pipette 1µl of ethanol onto one of the azide droplets. On the other azide droplet, pipette 1µl of 10% butanone solution in ethanol. The plate to be used for the memory test is now ready.

[0053] Subsequently, approximately 300 insects to be tested were released, specifically by dropping sodium azide droplets (containing either butanol or ethanol) approximately 4.5 cm away from each of the agar plate's lower or upper edges, taking the middle of the plate as a starting point. If the insects remembered that the bait was marked with butanol, they would move towards the butanol droplets, but would become immobilized there due to the presence of azide. If the insects did not remember, a similar number of animals would move towards the ethanol droplets as they did towards the butanol droplets. This also occurs if the animals were not trained with butanol. After allowing the animals to explore the plates containing butanol and ethanol for one hour, memory was quantified using the chemotactic index. The chemotactic index is calculated by subtracting the number of animals in the ethanol spots from the number of animals in the butanol spots and dividing by the total number of animals. Figure 5 shows the results of the memory test, expressing memory as the chemotactic index, taking into account the respective treatments for CL2122 and CL2355 animals.

[0054] conclusion Controlled experiments conducted showed that metformin and AP did not increase the normal chemotactic index of either trained or untrained CL2122 animals. This indicates that the normal memory of the control animals was not affected by either treatment. In CL2355, the chemotactic index was close to zero for animals trained with butanone. This indicates that the memory of these animals was almost completely lost. Treatment of insects with 111, 333, and 1000 IU / drop of AP statistically and dose-dependently increased the memory of CL2355 animals compared to buffered controls. Metformin-treated animals also showed a statistically significant increase in memory in this study. Therefore, treatment with AP counteracts memory loss in a dose-specific manner. The effect of AP on memory loss is somewhat less than that of metformin, which is known to act on neurodegeneration. This difference is likely due to the fact that insects absorb metformin within 48 hours, while AP is not absorbed and is excreted within a few minutes. After treatment with the test substance (AP or metformin), the entire learning process and memory test section is performed without these test substances. This test duration is approximately 3 hours, and during this period, metformin is more likely to be absorbed into the body, allowing it to last longer and provide a longer-lasting protective effect compared to AP.

Claims

1. A pharmaceutical composition comprising alkaline phosphatase for use in the treatment or prevention of a mammal suffering from or at risk of suffering from a neurodegenerative disorder caused by a decrease in peroxisome proliferator-activated receptor gamma coactivator 1 (PGC1) activity compared to a healthy individual, wherein the treatment comprises administering to the mammal a therapeutically effective amount of alkaline phosphatase that inhibits neurodegeneration by preventing a decrease in PGC1 activity, and the neurodegenerative disorder is Parkinson's disease.

2. A pharmaceutical composition comprising alkaline phosphatase for use according to claim 1, wherein preventing the decrease in PGC1 activity involves increasing the activation of adenosine monophosphate-activated protein kinase (AMPK) by alkaline phosphatase, thereby promoting an anti-stress response.

3. A pharmaceutical composition comprising alkaline phosphatase for use according to claim 1 or 2, wherein the therapeutically effective dose is 6 to 1350 U / day / kg.

4. A pharmaceutical composition comprising an alkaline phosphatase for use according to any one of claims 1 to 3, wherein the alkaline phosphatase is a tissue-specific ectophosphatase selected from the group consisting of intestinal AP (IAP), placental ALP (PALP), and hepatic AP (LAP).

5. A pharmaceutical composition comprising alkaline phosphatase for use according to any one of claims 1 to 4, wherein the treatment includes intravenous, parenteral, or oral administration.

6. A pharmaceutical composition comprising an alkaline phosphatase for use according to any one of claims 1 to 5, wherein the alkaline phosphatase is a recombinant alkaline phosphatase.

7. A pharmaceutical composition comprising an alkaline phosphatase for use according to any one of claims 1 to 6, wherein the alkaline phosphatase is one or more selected from the group consisting of biologically active fragments or derivatives of alkaline phosphatase, synthetic alkaline phosphatase derivatives, and chemically-pharmaceutically small molecules that exhibit functional alkaline phosphatase activity.

8. A pharmaceutical composition comprising alkaline phosphatase for use according to any one of claims 1 to 7, wherein the treatment or prevention includes delaying the onset of the neurodegenerative disorder, or reducing or preventing its progression.

9. A pharmaceutical composition comprising alkaline phosphatase for use according to any one of claims 1 to 8, wherein the treatment comprises reducing the inflammatory response in a mammal suffering from a neurodegenerative disorder.

10. Use of an alkaline phosphatase or a pharmaceutical composition comprising an alkaline phosphatase according to any one of claims 1 to 9 for the preparation of a pharmacopoeia for the prevention of a neurodegenerative disorder in a mammal at risk due to reduced PGC1 activity compared to a healthy individual, wherein the neurodegenerative disorder is Parkinson's disease.

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