Probiotic cocktail medical food composition and its use to slow the progression of Parkinson's disease

Abstract

The present invention provides a composition that can be manufactured into a medical food formulation and applied to improve or alleviate pathological changes associated with Parkinson's disease, such as motor impairment, cognitive degeneration, inflammatory responses, and mitochondrial dysfunction, and can be used for daily nutritional supplementation and to slow disease progression in Parkinson's disease patients or high-risk groups. [Solution] The present invention relates to a probiotic cocktail medical food composition and its use in slowing the progression of Parkinson's disease, wherein the composition comprises (a) at least one probiotic having neuroprotective or metabolic modulating function, and (b) a combination of functional amino acids including four or more essential amino acids.

Description

Technical Field

[0001] The present invention relates to a nutritional composition having a neuroprotective function, particularly a cocktail-type medical food composition containing probiotics and a combination of functional amino acids, and its use for delaying the progression of Parkinson's disease, belonging to the technical field of medical foods and nutritional compositions for the prevention and improvement of neurodegenerative diseases.

Background Art

[0002] Parkinson's disease (PD) is a central nervous system degenerative disease, and the main pathological feature is the gradual degeneration and loss of dopaminergic neurons. Clinically common symptoms include bradykinesia, muscle rigidity, resting tremors, and postural instability, and may be accompanied by cognitive impairment and mood abnormalities. The current treatment methods mainly focus on symptom control with drugs, especially L-DOPA replacement therapy is the most common, but long-term use is likely to cause side effects such as fluctuations in treatment effects and dyskinesia, and cannot fundamentally delay the progression of neurodegenerative diseases. Therefore, developing nutritional intervention strategies with preventive and disease-modifying potential has already become an important and urgent direction of adjuvant treatment.

[0003] According to recent research, the gut microbiota and gut immune function are closely related to the development of PD, and some probiotic strains have potential neuroprotective effects such as the regulation and control of neuroinflammation, antioxidant stress, and maintenance of intestinal barrier function. Probiotics can indirectly participate in the stability of the nervous system by affecting the production of short-chain fatty acids and immunometabolism. However, there are many existing technologies that focus on the effects of single probiotic strains on gut health, and there is still a lack of specific evidence for their application to Parkinson's disease.

[0004] Essential amino acids participate in important physiological processes such as neurotransmitter synthesis, energy metabolism, and muscle protein maintenance, and therefore possess potential physiological functions such as neuromodulation, anti-inflammatory effects, and support for mitochondrial function. In particular, compositions containing various essential amino acids can provide multiple metabolic support pathways and have the potential to be developed into disease-intervention nutritional prescriptions. Existing studies have rudimentarily investigated the supportive effects of such amino acids on neurological function, but they are often presented in the form of general nutritional supplements, and there is a lack of evaluation of combinations and functional verification for specific disease models. In particular, for chronic neurodegenerative diseases such as Parkinson's disease, there is still a lack of mechanism verification and application positioning for systematically investigating specific essential amino acid compositions, and this remains a technological gap.

[0005] Many conventional drug or nutritional interventions target a single target or mechanism of action, resulting in limited effectiveness in controlling disease progression in chronic neurodegenerative diseases with multiple pathogenic mechanisms, such as Parkinson's disease (PD). Recent research suggests that integrating multiple components with different physiological functions and pathways of action, known as "cocktail therapy," can simultaneously regulate and control multiple pathological mechanisms, further enhancing overall improvement and the potential to slow disease progression. While this strategy has already been applied in various fields such as antiviral, cancer, and metabolic diseases, nutritional cocktail compositions designed specifically for Parkinson's disease are still rare. There is a particular lack of complete intervention strategies that have a clear indication, are applicable to medical foods, and whose multiple biological mechanisms of action have been verified in animal models; further development is needed.

[0006] Prior art has focused on combinations of probiotics with herbal medicines, coenzymes, or other nutrients, often applied to general health purposes such as improving physical strength, boosting metabolism, or enhancing cardiovascular function, and rarely proposes applications with clear indications for neurodegenerative diseases such as Parkinson's disease. While some studies or technologies mention the potential influence of probiotics on the central nervous system by modulating vagus nerve or intestinal function, many of the relevant combinations do not contain tryptophan and branched-chain amino acids, and there is a lack of specific data to systematically verify therapeutic effects in animal models of Parkinson's disease, and there is still no clear evidence that they can slow disease progression. Thus, nutritional cocktail formulations for PD still belong to an unsatisfied technological need, and the compositions proposed in this invention possess clear innovation and applicability differences. [Overview of the Initiative] [Problems that the invention aims to solve]

[0007] This invention proposes a probiotic cocktail medical food composition that combines at least one probiotic with neuroprotective or metabolic modulating function with a combination of functional amino acids. Through the synergistic action of these two components, it modulates and controls the pathological mechanisms associated with Parkinson's disease from multiple angles, including improvement of motor and cognitive behavioral disorders, protection of dopaminergic neurons, reduction of neuroinflammation and oxidative pressure, restoration of mitochondrial function in the brain and muscles, and restructuring of the gut microbiota. The related effects have already been validated in multiple layers through 6-OHDA-induced PD animal models, confirming its potential to slow the progression of PD and its suitability as a long-term nutritional intervention prescription. [Means for solving the problem]

[0008] The primary objective of the present invention is to provide a probiotic cocktail medical food composition designed for the progression of Parkinson's disease, comprising a combination of at least one probiotic having neuroprotective or metabolic modulating function and functional amino acids, which, through synergistic action, modulate and control the pathological mechanisms associated with Parkinson's disease from multiple perspectives, further improving motor and non-motor symptoms, protecting dopaminergic neurons, alleviating neuroinflammation and oxidative pressure, improving mitochondrial function, and restructuring the gut microbiota. The composition is suitable as a nutritional intervention with disease-modifying potential, can be applied to daily supplementation for Parkinson's disease patients or high-risk groups, and provides a safe, long-term-use, and multi-target modulating adjunctive therapy option.

[0009] To achieve the above-mentioned objectives, the present invention provides a composition comprising (a) at least one probiotic having a neuroprotective or metabolic modulating function, and (b) a combination of functional amino acids comprising four or more essential amino acids.

[0010] To achieve the above-mentioned objectives, the probiotics having neuroprotective or metabolic modulating functions are selected from the group consisting of Lactobacillus, Lactococcus, Streptococcus, Bifidobacterium, Pediococcus, and Enterococcus genera.

[0011] To achieve the above object, the Lactobacillus genus is selected from the group consisting of L. acidophilus, L. antri, L. brevis, L. casei, L. coleohominis, L. crispatus, L. curvatus, L. fermentum, L. gasseri, L. johnsonii, L. mucosae, L. pentosus, L. plantarum, L. reuteri, L. rhamnosus, L. sakei, L. salivarius, L. paracasei, L. kisonensis, L. paralimentarius, L. perolens, L. apis, L. ghanensis, L. dextrinicus, L. shenzenensis and L. harbinensis.

[0012] To achieve the above object, the probiotics are live bacteria, dead bacteria, pasteurized bacteria, decomposition products, cell-free supernatants, cell wall extracts, exopolysaccharides or their immunostimulatory components.

[0013] To achieve the above object, the essential amino acids are selected from the group consisting of leucine, isoleucine, valine, lysine, methionine, phenylalanine, tryptophan, threonine and histidine.

[0014] To achieve the above object, the composition is a pharmaceutical composition, a nutritional supplement or a health food.

[0015] To achieve the above object, the composition contains a pharmaceutically acceptable carrier, excipient, diluent.

[0016] Another object of the present invention is to provide the use of the composition for preparing a drug for delaying the progression of Parkinson's disease.

[0017] To achieve the above-mentioned objectives, the non-motor symptoms associated with Parkinson's disease include, but are not limited to, cognitive impairment, memory impairment, impaired directionality, intellectual decline, executive function impairment, irritability, altered social behavior, anxiety, mood swings, depression, apathy, sleep disturbances, autonomic dysfunction, olfactory dysfunction, abnormal pain perception, or fatigue.

[0018] To achieve the above-mentioned objectives, the autonomic nervous system dysfunction includes constipation, hypotension, urinary dysfunction, or sweating abnormalities.

[0019] To achieve the above-mentioned objectives, the motor symptoms associated with Parkinson's disease include tremors, muscle rigidity, delayed movement, gait instability, limping, non-autonomous movement disorders, facial expression disorders, and ocular dysfunction.

[0020] To achieve the above-mentioned objectives, the non-autonomous motor abnormalities include difficulty with movement and muscle hypotonia.

[0021] To achieve the above-mentioned objectives, the facial expression disorders include mask-like facies and facial paralysis.

[0022] To achieve the above-mentioned objectives, the ocular dysfunction includes inability to open the eyelids, blepharospasm, decreased tear film secretion, and evaporative dry eye resulting from the blink reflex.

[0023] To achieve the above-mentioned objectives, the composition is used for daily nutritional supplementation in patients with Parkinson's disease or those at high risk of Parkinson's disease to slow disease progression. [Effects of the Invention]

[0024] Overall, the probiotic cocktail medical food composition disclosed by the present invention can be applied to the preparation of nutritional combination preparations for delaying the progression of Parkinson's disease, and can also be applied to the daily nutritional supplementation of Parkinson's disease patients or high-risk groups. Its application effects include improvement of motor dysfunction, improvement of cognitive performance, protection of neurons, reduction of neuroinflammatory response and antioxidant stress, and restoration of mitochondrial energy metabolism and intestinal flora balance, showing potential for systematic disease modification. It is a non-drug nutritional intervention strategy with technological innovation and clinical application value, which can act on multiple mechanisms, multiple targets and safely. It can be expected as an auxiliary solution for early intervention and delaying disease progression of Parkinson's disease, and also has the potential to be applied in the fields of prevention and supportive treatment of other neurodegenerative diseases.

Brief Description of Drawings

[0025] [Figure 1] It is a flowchart of an animal experiment of a PD-induced model. [Figure 2] Behavioral evaluation of the motor function of Parkinson's disease model rats: (A) Apomorphine rotation test in the second week, (B) Apomorphine rotation test in the tenth week, (C) Rotarod test, (D) Grip strength test. [Figure 3] Morris water maze spatial learning and memory test: (A) Test of swimming trajectory diagram, (B) Statistics of escape latency, (C) Stay time in the target quadrant and total swimming distance. [Figure 4] It is immunohistochemical staining of TH-positive dopaminergic neurons in the substantia nigra pars compacta and cell count statistics. [Figure 5] Analysis of gene expression related to inflammation and antioxidant in the striatal region: (A) Tnf-α, (B) Il-6, (C) Gpx, (D) Cat. [Figure 6] Analysis of brain mitochondrial function indexes: (A) OCR, (B) ECAR, (C) Sirt1 gene expression, (D) Pgc-1α gene expression. [Figure 7] Analysis of muscle mitochondrial function indexes: (A) OCR, (B) ECAR, (C) Sirt1 gene expression, (D) Pgc-1α gene expression. [Figure 8]This is an analysis of the relative abundance of the gut microbiota. [Figure 9] This is the result of the LEfSe analysis. [Modes for carrying out the invention]

[0026] All technical and scientific terms used herein have meanings that are commonly understood by those skilled in the art, unless otherwise specified. The materials used in this invention are commercially available materials unless otherwise specified.

[0027] The terms “one” or “one type” do not exclude multiple things; that is, unless the meaning is clearly expressed or required otherwise, a single form of “one,” “one type,” and “the said” should be understood to include multiple subjects. In other words, unless particularly explicitly stated or the semantic of a reference clearly implies the opposite, a single characteristic or limitation referred to in the entire text of the invention should include the corresponding multiple characteristics or limitations, and vice versa. Thus, unless specifically defined, the terms “one,” “one type,” and “the said” are synonymous with “at least one” or “one or more.” For example, when referring to “one type of component,” this includes mixtures of multiple components, etc.

[0028] The terms “approximately” or “about” are acceptable in the food or pharmaceutical industries to compensate for variations in food or medicine, such as differences in content due to manufacturing variations and / or product degradation over time. Allowing any actual changes in food or medicine using the aforementioned terms should be considered in light of the fact that the biometric properties in the subject are equivalent to the enumerated merits requiring the protection of the product.

[0029] The term "composition" refers to any form of composition, and multiple specific components may be selected to be included in the composition along with any further components.

[0030] The term "includes" and similar meanings should be interpreted in an open and inclusive manner as "includes, but does not limit."

[0031] The terms "one embodiment," "a single embodiment," and "one specific embodiment" indicate that one unique feature, attribute, or characteristic, or a group or set of unique features, attributes, or characteristics, are presented in combination with the individual meanings of the present invention in at least one embodiment of the invention. Such meanings appearing throughout this patent specification do not necessarily refer to the same embodiment. These unique features, attributes, or characteristics may be combined in any appropriate manner in one or more embodiments.

[0032] The term "prevention" refers to preventing a disease, disease state, or symptoms, and, after the etiology, disease state, or symptoms of the disease have been improved or eliminated, the need to prevent further increase and spread of recurrence or progression.

[0033] The term "treatment" refers to a therapeutic intervention that can effectively treat a disease, condition, or symptoms, but it can also mean improvement, advancement, control, suppression of progression, prevention of progression, and prevention of recurrence.

[0034] The term "probiotics" refers to microorganisms that contain live microorganisms, that is, microorganisms capable of reproduction. Probiotics are known as microorganisms that, when administered in appropriate amounts based on the technology available at the time of the patent application, provide health benefits to the host.

[0035] The term "probiotic cocktail medical food composition" refers to a complex medical food comprising at least one probiotic and a nutrient with specific physiological functions (e.g., essential amino acids). The composition possesses multiple biological activities and can achieve effects that slow the progression of Parkinson's disease through different mechanisms (e.g., neuroprotection, anti-inflammatory, antioxidant, mitochondrial activation, and gut microbiota regulation). The probiotic cocktail composition as used in this invention is a complex medical nutritional formulation based on a multi-component cooperative mechanism, applicable to adjunctive intervention for neurodegenerative diseases and daily nutritional supplementation.

[0036] The term "probiotics having neuroprotective or metabolic modulatory functions" refers to microbial strains that can modulate neuroinflammation, antioxidant activity, mitochondrial function, or gut microbiota through direct or indirect mechanisms, and that also have protective or stabilizing effects on the nervous system. The probiotics include live bacteria, dead bacteria, degradation products, bacterial supernatant, extracellular polysaccharides, or components having immunomodulatory functions.

[0037] The term "essential amino acid" refers to alpha-amino acids that the human body cannot synthesize on its own, or whose synthesis rate is insufficient to meet physiological needs, and which must be obtained through diet or exogenous supplementation. According to a general nutritional definition, common essential amino acids include leucine, isoleucine, valine, lysine, methionine, phenylalanine, tryptophan, threonine, and histidine.

[0038] The term "slowing the progression of Parkinson's disease" refers to slowing or inhibiting the worsening of the disease's progression through some form of intervention, and includes, but is not limited to, improvement of motor function impairment, reduction of non-motor symptoms, maintenance of neuronal integrity, or improvement of quality of life.

[0039] The term "non-motor symptoms associated with Parkinson's disease" includes, but is not limited to, cognitive impairment, memory impairment, impaired directionality, intellectual decline, impaired executive function, irritability, changes in social behavior, restlessness, mood swings, depression, apathy, sleep disturbances, autonomic dysfunction (e.g., constipation, hypotension, urinary dysfunction, abnormal sweating), decreased olfactory function, abnormal pain perception, and fatigue.

[0040] The term "motor symptoms associated with Parkinson's disease" includes, but is not limited to, tremors, muscle rigidity, delayed movement, gait instability, limping, non-autonomous motor disorders (e.g., dyskin, hypotonia), facial expression disorders (e.g., masked face, facial paralysis), and ocular dysfunction (e.g., inability to open eyelids, blepharospasm, decreased tear film secretion, evaporative dry eye due to blink reflex).

[0041] The term "multi-target" refers to an intervention strategy that simultaneously acts on two or more physiological or molecular mechanisms related to disease pathology, such as simultaneously regulating neuroinflammatory responses, mitochondrial function, antioxidant capacity, neurotransmission regulation, and gut microbiota balance. Compared to single-target interventions, multi-target strategies can enhance the overall benefit of the intervention and contribute to multi-layered regulatory control of disease progression.

[0042] The Lactobacillus reuteri Y7 used in this embodiment is deposited with the Center for Biological Resource Conservation and Research (BCRC) in Taiwan, with deposit number BCRC911162. The aforementioned strain is also deposited with the National Institute of Technology and Evaluation (NITE) Patent Microbial Depository Center (Room 120, 2-5-8 Kazusa-Kamatari, Kisarazu City, Chiba Prefecture, 292-0818 Japan), an international depositary organization in Japan, with deposit number NITE BP-04078.

[0043] The present invention relates to a complex nutritional composition, whose design concept is based on the concept of "cocktail therapy." It integrates a combination of probiotics and functional amino acids having neuroprotective or metabolic modulating functions, and regulates and controls multiple physiological and pathological mechanisms associated with the progression of Parkinson's disease through the cooperative and complementary effects of multiple components. The composition may contain probiotics of one or more origins, in the form of live bacteria, dead bacteria, treated bacterial cells or their derivatives, and may also contain nutrient components, such as essential amino acids, that have potential for regulating neurotransmission, energy metabolism, and anti-inflammatory effects. These can be combined in specific proportions and manufactured into oral dosage forms such as powders, tablets, capsules, and beverages. The composition can be used as a pharmaceutical composition, nutritional supplement, or health food, and its target population covers Parkinson's disease patients or high-risk groups. It can be used for daily nutritional supplementation or as an adjunct intervention to delay disease progression, and has potential disease-modifying effects. Its applications include improving motor function impairment, cognitive degeneration, neuroinflammation, and mitochondrial dysfunction, and it has the potential to regulate and control the health of the gut microbiota and central nervous system through the gut-brain axis mechanism, making it an innovative nutritional intervention with clinical potential.

[0044] In one embodiment, the probiotics include, but are not limited to, any species or mixtures of any of the genera Lactobacillus, Bifidobacterium, Streptococcus, Lactococcus, Pediococcus, and Enterococcus, and the bacterial cells include, but are not limited to, live, dead, pasteurized, degradation products, extracellular polysaccharides, or bacterial supernatant. Any of the above-mentioned probiotics may affect nerve function, and the Lactobacillus reuteri Y7 strain used in the embodiments of the present invention is a preferred embodiment.

[0045] In one embodiment, the essential amino acids are selected from leucine, isoleucine, valine, lysine, methionine, phenylalanine, tryptophan, threonine, and histidine, and can be used individually or in combination depending on different functional needs.

[0046] In one embodiment, the improved motor symptoms may include tremors, rigidity, delayed movement, gait abnormalities, delayed facial expression, eyelid dysfunction, and non-autonomous movement abnormalities. The non-motor symptoms may cover expressions such as anxiety, depression, sleep disorders, gastrointestinal motility abnormalities, and dry eyes.

[0047] In one embodiment, the composition is manufactured into a pharmaceutical composition, nutritional supplement, or health food, and is prepared in the form of a powder, tablet, capsule, or beverage by incorporating pharmaceutically acceptable carriers, excipients, and diluents, which are advantageous for clinical application and long-term intake.

[0048] Example 1: The probiotic cocktail medical food composition showed an improvement effect on Parkinson's disease model rats.

[0049] This example aims to investigate the effect of a probiotic cocktail medical food composition (hereinafter abbreviated as "TMU-01 formulation") containing a specific probiotic (Lactobacillus reuteri Y7) and a combination of functional amino acids on the progression of Parkinson's disease. In the experiment, a 6-OHDA-induced rat Parkinson's disease model was constructed using a typical PD model preparation method, and as shown in Figure 1, the neurotoxin 6-hydroxydopamine (6-OHDA) was injected unilaterally into the medial forebrain bundle (MFB) of rats to induce damage to dopaminergic neurons.

[0050] [1] I. Animal grouping and processing methods The experiment used 18 Sprague-Dawley rats, which were randomly divided into three groups (n=6 in each group): • ND group: This was the normal control group, which did not receive 6-OHDA injections but received forced oral administration with physiological saline. • PD group: This group consisted of Parkinson's disease models who received injections of 6-OHDA into the right MFB followed by forced oral administration of saline solution. • Group M: This was the treatment group, and the model was constructed using the same method as the PD group. The TMU-01 prescription was administered orally daily and irrigated into the stomach. The experiment was conducted during the following time slots (as shown in Figure 1): • Administer a 6-OHDA injection in week 0; • Apomorphine-induced rotational testing will be performed in weeks 2 and 10. • Conduct the rotor rod test, grip strength test, and Morris water maze test in weeks 8-9; • Animals were sacrificed in the 10th week for tissue staining and subsequent analysis.

[0051] 2. Assessment of Behavioral Function

[0052] Apomorphine-induced rotational behavior tests were conducted in weeks 2 and 10. The results showed that the PD group had significantly higher rotational counts than the ND group (as shown in Figures 2A and 2B) in both tests, indicating a clear impairment of their motor function. The M group had significantly lower rotational counts than the PD group, demonstrating that this prescription can effectively improve rotational abnormalities.

[0053] In the rotor rod test, the PD group rats showed a significantly shorter dwell time on the platform, representing a decline in their motor coordination function. The M group, on the other hand, showed significantly better performance in this index than the PD group (as shown in Figure 2C), indicating a certain degree of recovery in their motor stability and balance ability.

[0054] Based on the overall behavioral assessment, this prescription significantly improved motor impairment in a 6-OHDA-induced Parkinson's disease rat model, including reduced rotational behavior, improved motor coordination and muscle strength expression, and demonstrated potential neuroprotective and motor function recovery effects.

[0055] 3. Cognitive function assessment

[0056] To evaluate the impact of this prescription on cognitive function, the Morris water maze test was administered during weeks 8-9 to assess spatial learning and memory abilities.

[0057] As shown in Figures 3A to 3C, the PD group rats exhibited a significantly prolonged escape latency period, a clearly reduced time spent in the target quadrant, and an increased total swimming distance, indicating a clear impairment of their spatial learning ability and memory retrieval ability.

[0058] In contrast, the M group rats were significantly superior to the PD group in indicators such as escape latency and time spent in the target area, demonstrating that this prescription can effectively improve spatial learning and memory abilities in Parkinson's disease, and that their cognitive expression was close to that of the ND group, indicating that this prescription can improve cognitive impairment in Parkinson's disease.

[0059] Overall, according to the Morris water maze results, this prescription has a significant protective effect on spatial learning and memory abilities in the 6-OHDA-induced PD model and helps to slow cognitive degeneration associated with Parkinson's disease.

[0060] IV. Neuron protective effect

[0061] To evaluate the protective effect of this formulation on dopaminergic neurons in the substantia nigra region of the midbrain, animals were sacrificed at week 10, brain samples were collected, and tyrosine hydroxylase (TH) immunohistochemical staining was performed to analyze the distribution and number of positive neurons in the substantia nigra pars compacta (SNpc).

[0062] As shown in Figures 4A and 4B, the number of TH-positive cells in the SNpc region of PD rats was significantly lower than that of the ND group, indicating a loss of dopaminergic neurons and consistent with the neuropathological characteristics of Parkinson's disease.

[0063] The number of TH-positive cells in the M group rats was significantly preserved, with a significantly higher number of positive cells compared to the PD group. This indicates that neuronal damage and loss can be effectively suppressed through intervention with this formulation, demonstrating its dopamine neuroprotective effect.

[0064] These results demonstrate that this prescription can improve behavioral and cognitive expression, significantly preserve dopamine neurons at the tissue level, and significantly increase the number of TH-positive cells. This indicates that the prescription has neuroprotective effects and the potential to slow the pathological progression of Parkinson's disease.

[0065] V. Analysis of inflammation and antioxidant indicators

[0066] To further investigate the modulating effects of this prescription on neuroinflammation and oxidative pressure, rat striatum tissue was collected in week 10, and analyses were performed to enhance the mRNA expression levels of genes related to inflammation and antioxidant activity.

[0067] As shown in Figures 5A and 5B, the expression of the inflammatory factors tumor necrosis factor-α (Tnf-α) and interleukin-6 (Il-6) in PD group rats was significantly elevated compared to the normal control ND group, indicating an increase in the neuroinflammatory response in the Parkinson's disease model. In contrast, the expression levels of the inflammatory factors Tnf-α and Il-6 in M ​​group rats were significantly lower than in the PD group, demonstrating a clear anti-inflammatory effect of this formulation.

[0068] As shown in Figures 5C and 5D, the expression levels of the antioxidant enzymes glutathione peroxidase (Gpx) and catalase (Cat) were significantly lower in the PD group than in the ND group, indicating damage to their antioxidant system. Gpx and Cat gene expression in the M group rats was remarkably higher than in the PD group, demonstrating that this formulation enhances the antioxidant capacity of cells and strengthens the defense mechanisms against oxidative pressure in nerve cells.

[0069] In summary, this formulation possesses a dual effect: simultaneously reducing the expression of neuroinflammatory factors and enhancing the activity of antioxidant enzyme genes, thereby regulating neuroinflammation and oxidative pressure, contributing to neuroprotection and delaying neurodegeneration.

[0070] VI. Analysis of Mitochondrial Function

[0071] To evaluate the effects of this formulation on mitochondrial function, rat striatal and soleus muscle tissues were collected, and metabolic function-related indicators were analyzed, including oxygen consumption rate (OCR), extracellular acidification rate (ECAR), and the expression levels of Sirtuin 1 (Sirt1) and peroxisome proliferator-activated receptor gamma coactivator 1-alpha (Pgc-1α), genes related to mitochondrial biosynthesis.

[0072] As shown in Figures 6A and 6B, OCR and ECAR in the striatum of PD rats were significantly lower than those of the ND group, indicating mitochondrial dysfunction. Mitochondrial function in the M group was significantly restored, with both OCR and ECAR being significantly higher than in the PD group, demonstrating a clear improvement in mitochondrial function.

[0073] Further analysis of genes related to mitochondrial synthesis revealed a clear decrease in the expression levels of Sirt1 and Pgc-1α in the PD group, while the expression levels in the M group rats were significantly increased (as shown in Figures 6C and 6D), further supporting the restoration and enhancement of mitochondrial function.

[0074] Similar results were observed in muscle tissue. As shown in Figures 7A to 7D, in the soleus muscle, OCR, ECAR, Sirt1, and Pgc-1α all decreased in the PD group, while the M group showed significant recovery. This indicates that the formula not only acts on the central nervous system but also has a restorative effect on mitochondrial function in surrounding tissues, demonstrating that the formula can improve mitochondrial health systemically.

[0075] Overall, this formulation effectively improves nerve function damage caused by energy impairment in animal models of Parkinson's disease by restoring mitochondrial energy metabolism and enhancing mitochondrial production, demonstrating its potential for systematic mitochondrial regulation.

[0076] 7. Analysis of the intestinal microbiota

[0077] To evaluate the regulatory effect of this formulation on the composition of the gut microbiota, rat fecal samples were collected in the 10th week of the experiment, and 16S rRNA high-throughput sequencing analysis was performed, along with analysis of the relative abundance of bacterial groups and comparison using LEfSe statistics.

[0078] As shown in Figures 8A to 8C, the structure of the gut microbiota in PD rats underwent significant changes at the phylum, genus, and species levels. A decrease in the relative abundance of probiotics and an increase in various potential pathogens were observed, indicating that the PD model led to an imbalance in the gut microbiota.

[0079] The structure of the gut microbiota in group M rats differed significantly from that of the PD group. This demonstrated that the treatment restored the relative abundance of beneficial bacteria and increased microbial diversity, bringing the bacterial distribution closer to that of the ND group. This indicates that the treatment can reconstruct the damaged gut microecology in a Parkinson's disease model.

[0080] Furthermore, as shown in Figures 9A and 9B, we compared the differences in enteric flora among the three groups via LEfSe analysis and discovered that group M has distinct bacterial characteristics from group PD. This indicates that intervention with this formulation leads to an increase in the dominance of specific beneficial bacterial species, has a good regulatory effect on the composition of enteric flora, and can restore diversity and ecological balance of enteric microorganisms.

[0081] Based on the results described above, the TMU-01 formula has a beneficial regulatory function for the gut microbiota, can restore the diversity and ecological balance of gut microbiota, and can indirectly alleviate neurodegenerative symptoms associated with Parkinson's disease through the gut-brain axis mechanism, potentially providing a regulatory effect on systemic diseases.

[0082] [1] Based on the results of the embodiments of the present invention, the TMU-01 formulation (a cocktail medical food composition containing a combination of probiotics and functional amino acids) can exhibit multiple remarkable physiological improvement effects in an animal model of 6-OHDA-induced Parkinson's disease and has the following technical effects: 1. Improvement of motor function impairment: We demonstrated that apomorphine can effectively reduce apomorphine-induced rotational behavior, improve dwell time on the Rotarod platform and grip strength test performance, and restore motor function in PD model rats (as shown in Figure 2). 2. Improved cognitive function: It improves spatial learning and memory representation in the Morris water maze, shortens the escape latency period, increases the time spent in the target area, and demonstrates a protective effect on spatial memory (as shown in Figure 3). 3. Protection of dopamine neurons: The number of TH-positive cells in the SNpc region significantly increased, demonstrating a neuroprotective effect and slowing the degeneration of dopamine neurons (as shown in Figure 4). 4. Enhanced anti-inflammatory and antioxidant capacity: Reduces the expression of inflammatory factors Tnf-α and Il-6, and increases the expression of antioxidant-related genes Gpx and Cat, helping to stabilize the neural microenvironment (as shown in Figure 5). 5. Restoration of mitochondrial metabolic function: We demonstrated that it improves OCR and ECAR values ​​in the striatum and soleus muscle, promotes the expression of mitochondrial biosynthesis genes Sirt1 and Pgc-1α, and has the effect of restoring mitochondrial function and enhancing energy metabolism (as shown in Figures 6 and 7). 6. Regulate the gut microbiota and restore microecological equilibrium: Significantly improve the relative abundance of probiotics, alleviate the imbalance of enteric bacteria associated with Parkinson's disease, and confirm clear differences in bacterial flora indicators through LEfSe analysis (as shown in Figures 8 and 9).

[0083] Based on the results described above, this formulation possesses multiple regulatory mechanisms, can act simultaneously on the central nervous system and surrounding tissues, and exhibits remarkable improvement effects on behavioral function, neuropathology, molecular labeling, and systemic metabolism in animal models of Parkinson's disease. As a supplemental nutritional therapy with the potential to slow disease progression, this formulation is expected to have high clinical and industrial application value when applied to the fields of preventive health care, early intervention, or disease regulation for Parkinson's disease.

[0084] The innovative aspect of this invention lies in providing a medical food composition combining probiotics and functional amino acids to regulate and control key aspects of Parkinson's disease progression, such as neuronal degeneration, inflammatory responses, mitochondrial dysfunction, and cognitive degeneration. This combination possesses multiple mechanisms of action and functional complementarity, significantly outperforming existing single-component products, thereby slowing the progression of PD pathology from its source. This invention specifically targets Parkinson's disease as its indication, employing a 6-OHDA-induced Parkinson's disease animal model for systematic validation, covering multi-layered indicators including motor behavior, cognitive expression, dopamine neuron integrity, inflammation and antioxidant markers, mitochondrial function, and gut microbiota structure. Experimental results show that this composition significantly improves motor and memory function in Parkinson's disease model animals, protects dopaminergic neurons, reduces neuroinflammation, enhances antioxidant capacity, restores mitochondrial metabolic activity in brain and muscle tissue, and reconstructs the structure of the gut microbiota, primarily composed of beneficial bacteria. The effects described above are closely related to the progression of Parkinson's disease, indicating that this formulation is a non-pharmacological nutritional intervention strategy with disease-modifying potential. Unlike traditional drugs, which only alleviate symptoms, the medical food composition of this invention can serve as a long-term supportive, disease-modifying intervention and is applicable to daily nutritional supplementation for early-stage or high-risk Parkinson's disease patients. Furthermore, this composition can be manufactured in the form of powders, tablets, capsules, or beverages, making it convenient for long-term intake and clinical initiation. It can serve as an adjunct treatment option for Parkinson's disease patients, as well as for preventative nutritional fortification in the general population, possessing high clinical value and industrial application potential.

[0085] Biological material deposit information The Food Industry Development Research Institute, a public interest incorporated foundation, deposited the strain on November 29, 2022, with deposit number BCRC911162. The strain was also deposited at the National Institute of Technology and Evaluation (NITE) Patent Microorganism Depositary Center (2-5-8 Kazusa-Kamatari, Kisarazu City, Chiba Prefecture, 292-0818 Japan, Room 120), on February 9, 2024, with deposit number NITE BP-04078.

Claims

1. A composition comprising (a) a probiotic having at least one neuroprotective or metabolic modulating function, and (b) a combination of functional amino acids including four or more essential amino acids.

2. The composition according to claim 1, wherein the probiotic having neuroprotective or metabolic modulating function is selected from the group consisting of Lactobacillus, Lactococcus, Streptococcus, Bifidobacterium, Pediococcus, and Enterococcus genera.

3. The Lactobacillus genus is L. acidophilus, L. antri, L. Brevis, L. casei, L. coleohominis, L. crispatus, L. curvatus, L. fermentum, L. gasseri, L. Johnsonii, L. mucosae, L. pentosus, L. plantarum, L. reuteri, L. rhamnosus, L. sakei, L. salivarius, L. paracasei, L. kisonensis, L. paralimentarius, L. perolens, L. The composition according to claim 2, selected from the group consisting of L. apis, L. ghanenis, L. dextrinicus, L. shenzhenensis, and L. harbinensis.

4. The composition according to claim 1, wherein the probiotics are live bacteria, dead bacteria, pasteurized bacteria, degradation products, bacterial supernatant, cell wall extracts, extracellular polysaccharides or their immunostimulant components.

5. The composition according to claim 1, wherein the essential amino acids are leucine, isoleucine, valine, lysine, methionine, phenylalanine, tryptophan, threonine, and histidine.

6. The composition according to claim 1, wherein the composition is a pharmaceutical composition, a nutritional supplement, or a health food.

7. The composition according to claim 1, comprising a pharmaceutically acceptable carrier, an excipient, and a diluent.

8. The composition according to claim 1, wherein the composition is a powder, a tablet, a capsule, or a beverage.

9. An application for preparing a drug that slows the progression of Parkinson's disease, using the composition described in claim 1.

10. The use according to claim 9, wherein slowing the progression of Parkinson's disease is to alleviate motor symptoms and / or non-motor symptoms associated with Parkinson's disease.

11. The use of claim 10, which alleviates the non-motor symptoms associated with Parkinson's disease, including cognitive impairment, memory impairment, directionality impairment, intellectual decline, executive function impairment, irritability, changes in social behavior, anxiety, mood swings, depression, apathy, sleep disorders, autonomic nervous system dysfunction, olfactory dysfunction, abnormal pain perception, or fatigue.

12. The use according to claim 11, wherein the autonomic nervous system dysfunction includes constipation, hypotension, urinary dysfunction, or sweating abnormalities.

13. The use of claim 12, which alleviates the motor symptoms associated with Parkinson's disease, including tremor, muscle rigidity, delayed movement, gait instability, limping, non-autonomous movement disorders, facial expression disorders, and ocular dysfunction.

14. The use described in claim 13, wherein the non-autonomous motor abnormality includes difficulty with movement and muscle hypotonia.

15. The use according to claim 13, wherein the facial expression disorder includes a mask-like face and facial paralysis.

16. The use according to claim 13, wherein the ocular dysfunction includes inability to open the eyelids, blepharospasm, decreased tear film secretion, and evaporative dry eye resulting from the blink reflex.

17. The use of claim 9, which slows the progression of Parkinson's disease through improved anti-inflammatory capacity, improved antioxidant capacity, restoration of mitochondrial metabolic function, regulation of the gut microbiota, or reconstruction of the gut microbiological balance.

18. The composition is used for the daily nutritional supplementation of Parkinson's disease patients or high-risk groups to slow the progression of the disease, as described in claim 10.

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