Probiotic cocktail medical food composition and application thereof to postpone progress of Parkinson's disease

Abstract

The invention relates to a probiotic cocktail medical food composition and application thereof in delaying the progress of Parkinson's disease. The composition comprises (a) at least one probiotic with a neuroprotection or metabolism regulation function; (b) a functional amino acid combination, wherein the functional amino acid combination comprises four or more kinds of essential amino acid. The composition can be prepared into a medical food formula, is suitable for improving or relieving pathological changes such as dyskinesia, cognitive degeneration, inflammatory response and granulocyte function imbalance related to the Parkinson's disease, and can be used for daily nutrition supplement and disease progress delay of patients with the Parkinson's disease or high-risk groups.

Description

[0001] This application claims priority to U.S. Patent Application No. 63 / 705,596, filed on October 10, 2024, the entire contents of which are incorporated herein by reference. Technical Field

[0002] This invention relates to the field of nutritional composition technology, specifically to a probiotic cocktail medical food composition and its use in delaying the progression of Parkinson's disease. Background Technology

[0003] Parkinson's disease (PD) is a neurodegenerative disease characterized by the gradual degeneration and loss of dopaminergic neurons. Common clinical symptoms include bradykinesia, muscle rigidity, resting tremor, and postural instability, and may be accompanied by cognitive impairment and mood abnormalities. Current treatment primarily involves medication to control symptoms, with levodopa (L-DOPA) supplementation being the most common approach. However, long-term use can lead to fluctuating efficacy and side effects such as dyskinesia, and it cannot fundamentally delay the progression of neurodegeneration. Therefore, developing nutritional intervention strategies with preventative and disease-modifying potential has become an important and urgent direction for adjunctive therapy.

[0004] Recent studies have indicated that the gut microbiota and intestinal immune function are closely related to the development of Parkinson's disease (PD), and some probiotic strains have potential neuroprotective effects such as regulating neuroinflammation, resisting oxidative stress, and maintaining intestinal barrier function. Probiotics can indirectly participate in the stability of the nervous system by influencing the production of short-chain fatty acids and immune metabolism. However, existing technologies mostly focus on the benefits of single probiotic strains for gut health, and there is still a lack of concrete evidence for their application in Parkinson's disease.

[0005] Furthermore, essential amino acids, due to their involvement in crucial physiological processes such as neurotransmitter synthesis, energy metabolism, and muscle protein maintenance, possess potential physiological functions including neuroregulation, anti-inflammation, and mitochondrial function support. In particular, compositions containing multiple essential amino acids, offering support for multiple metabolic pathways, have the potential to be developed into disease-interventional nutritional formulations. While existing research has preliminarily explored the auxiliary benefits of these amino acids on neurological function, these studies have mostly been presented in the form of general nutritional supplements, lacking evaluation and functional validation of combinations designed for specific disease patterns. Especially for chronic neurodegenerative diseases such as Parkinson's disease, there is currently a lack of systematic exploration of the mechanistic validation and application positioning of specific essential amino acid compositions, representing a technological gap.

[0006] Traditional drug or nutritional interventions often target single targets or mechanisms of action, offering limited effectiveness in controlling the course of chronic neurodegenerative diseases with multiple pathogenic mechanisms, such as Parkinson's disease (PD). Recent research indicates that integrating multiple components with different physiological functions and pathways of action—the so-called "cocktail therapy"—can simultaneously regulate multiple pathological mechanisms, thereby enhancing overall improvement and potentially delaying disease progression. This strategy has been applied in various fields, including antiviral, cancer, and metabolic diseases. However, nutritional cocktail compositions designed specifically for Parkinson's disease are still rare, particularly lacking comprehensive intervention programs with clearly defined indications, applicability to medical foods, and validated multiple biological mechanisms of action through animal models. Further development is needed.

[0007] Existing technologies primarily focus on combining probiotics with traditional Chinese medicine, coenzymes, or other nutrients for general health purposes such as improving physical strength, promoting metabolism, or strengthening cardiovascular function. Few applications specifically target neurodegenerative diseases like Parkinson's disease. While some studies or technologies mention that probiotics may affect the central nervous system by regulating the vagus nerve or intestinal function, these combinations often lack tryptophan and branched-chain amino acids, and there is a lack of concrete data on systematic efficacy verification using animal models of Parkinson's disease. Therefore, there is no clear evidence to support that they can delay disease progression. Thus, a nutritional cocktail formulation for PD remains an unmet technical need, and the composition proposed in this invention possesses clear innovation and application differentiation. Summary of the Invention

[0008] The primary objective of this invention is to provide a probiotic cocktail medical food composition and its use in delaying the progression of Parkinson's disease. It relates to a probiotic cocktail medical food composition designed to target the progression of Parkinson's disease. This composition comprises at least one probiotic with neuroprotective or metabolic regulatory functions and a combination of functional amino acids. Through the synergistic effect of these two components, it regulates the pathological mechanisms associated with Parkinson's disease at multiple levels, thereby improving motor and non-motor symptoms, protecting dopaminergic neurons, reducing neuroinflammation and oxidative stress, enhancing mitochondrial function, and reshaping the gut microbiota structure. This composition is suitable as a nutritional intervention with disease-modifying potential and can be applied as a daily supplement for Parkinson's disease patients or high-risk groups, providing a safe, long-term usable adjunctive treatment option with multi-target regulatory capabilities.

[0009] To achieve the above objectives, the present invention provides a composition comprising (a) at least one probiotic having neuroprotective or metabolic regulatory functions; and (b) a functional amino acid combination comprising four or more essential amino acids.

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

[0011] To achieve the above objectives, the Lactobacillus genus is selected from L. acidophilus, L. antri, and L.

[0012] 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.

[0013] paralimentarius, L.perolens, L.apis, L.ghanensis, L.dextrinicus, L.

[0014] The group consisting of shenzenensis and L. harbinensis.

[0015] To achieve the above objectives, the probiotic may be a live bacterium, a dead bacterium, a bacterium that has undergone low-temperature sterilization, a lysis product, a bacterial supernatant, a cell wall extract, an extracellular polysaccharide, or an immunostimulatory component thereof.

[0016] To achieve the above objectives, the essential amino acid is selected from the group consisting of leucine, isoleucine, valine, lysine, methionine, phenylalanine, tryptophan, threonine, and histidine.

[0017] To achieve the above objectives, the composition is a pharmaceutical composition, a nutritional supplement, or a health food.

[0018] To achieve the above objectives, the composition includes a pharmaceutically acceptable carrier, excipient, and diluent.

[0019] To achieve the above objectives, the composition is in the form of powder, tablets, capsules, or beverage.

[0020] Another object of the present invention is to provide the use of the composition as described above for the preparation of a medicament for delaying the progression of Parkinson's disease.

[0021] To achieve the above objectives, the delay in the progression of Parkinson's disease is to reduce Parkinson's disease-related motor symptoms and / or reduce Parkinson's disease-related non-motor symptoms.

[0022] To achieve the above objectives, the reduction of nonmotor symptoms associated with Parkinson's disease includes cognitive impairment, memory impairment, disorientation, cognitive decline, executive dysfunction, irritability, changes in interpersonal behavior, anxiety, mood swings, depression, apathy, sleep disorders, autonomic dysfunction, olfactory dysfunction, abnormal pain perception, or fatigue.

[0023] To achieve the above objectives, the autonomic nervous system dysfunction may include constipation, low blood pressure, urinary disorders, or abnormal sweating.

[0024] To achieve the above objectives, the reduction of Parkinson's disease-related motor symptoms includes tremor, muscle rigidity, bradykinesia, gait instability, foot dragging, involuntary movement disorders, facial expression disorders, and eye dysfunction.

[0025] To achieve the above objectives, the involuntary movement disorder includes difficulty in movement and insufficiency of muscle tone.

[0026] To achieve the above objectives, the facial expression disorder includes mask face and facial paralysis.

[0027] To achieve the above objectives, the ocular dysfunction includes inability to open or close the eyelids, blepharospasm, reduced tear film secretion, and evaporative dry eye caused by non-blinking reflex.

[0028] To achieve the above objectives, delaying the progression of Parkinson's disease can be achieved by enhancing anti-inflammatory capabilities, enhancing antioxidant capabilities, restoring mitochondrial metabolic function, adjusting the gut microbiota, or rebuilding the gut microecological balance.

[0029] To achieve the above objectives, the composition is used as a daily nutritional supplement for patients with Parkinson's disease or high-risk groups to delay disease progression.

[0030] In summary, the probiotic cocktail medical food composition disclosed in this invention can be used to prepare nutritional formulations for delaying the progression of Parkinson's disease, and can also be used as a daily nutritional supplement for Parkinson's patients or high-risk groups. Its benefits include improving motor dysfunction, enhancing cognitive performance, protecting neurons, reducing neuroinflammatory responses and oxidative stress, and restoring mitochondrial energy metabolism and gut microbiota balance, demonstrating its potential to modify systemic diseases. It possesses technological innovation and clinical application value, representing a multi-mechanism, multi-target, safe, and accessible non-pharmacological nutritional intervention strategy. It is expected to serve as an adjunct solution for early intervention and disease progression delay in Parkinson's disease, and also has the potential to be extended to the prevention and supportive treatment of other neurodegenerative diseases.

[0031] Therefore, this invention proposes a probiotic cocktail medical food composition, combining at least one probiotic with neuroprotective or metabolic regulatory functions and a functional amino acid combination. Through the synergistic effect of the two, it regulates the pathological mechanisms related to Parkinson's disease from multiple perspectives, including improving motor and cognitive behavioral disorders, protecting dopaminergic neurons, reducing neuroinflammation and oxidative stress, restoring mitochondrial function in the brain and muscles, and remodeling the gut microbiota structure. The related efficacy has been validated at multiple levels using a 6-OHDA-induced PD animal model, confirming its potential to delay PD progression and its suitability as a long-term nutritional intervention formulation. Attached Figure Description

[0032] Figure 1 Flowchart of PD-induced animal model experiments.

[0033] Figure 2 Motor function and behavioral assessment of rats with Parkinson's disease model: (A) Apomorphine rotation test at week 2; (B) Apomorphine rotation test at week 10; (C) Rotarod balance test; (D) Grip strength test.

[0034] Figure 3 Morris Water Maze Spatial Learning and Memory Test: (A) Test swimming trajectory map; (B) Escape latency statistics; (C) Target quadrant dwell time and total swimming distance.

[0035] Figure 4 Immunohistochemical staining and cell count of TH-positive dopaminergic neurons in the substantia nigra compacta.

[0036] Figure 5 Analysis of the expression of genes related to inflammation and antioxidant activity in the striatum: (A) Tnf-α; (B) Il-6; (C) Gpx; (D) Cat.

[0037] Figure 6Analysis of mitochondrial functional indicators in the brain: (A) OCR; (B) ECAR; (C) Sirt1 gene expression; (D) Pgc-1α gene expression.

[0038] Figure 7 Analysis of functional indicators of myomitochondria: (A) OCR; (B) ECAR; (C) Sirt1 gene expression; (D) Pgc-1α gene expression.

[0039] Figure 8 This study analyzed the relative abundance of gut microbiota.

[0040] Figure 9 LEfSe analysis results Detailed Implementation

[0041] All technical and scientific terms used in this specification, unless otherwise defined, have the meanings commonly understood by one of ordinary skill in the art; the materials used in this invention, unless otherwise specified, are commercially available and readily available.

[0042] The terms "a" or "an" do not exclude a plurality; that is, unless the meaning clearly indicates or requires otherwise, the single-form terms "a," "an," and "the" should be understood to include a plurality of objects. In other words, unless otherwise expressly stated or the meaning of the reference clearly implies the opposite, references to a single characteristic or limitation throughout this invention should include the corresponding plurality of characteristics or limitations, and vice versa. Therefore, unless otherwise defined, the terms "a," "an," "the," "at least one," or "one or more" have the same meaning. For example, when referring to "an ingredient," it includes a mixture of multiple ingredients, etc.

[0043] The terms "about" or "approximately" allow for variations permitted in the food or pharmaceutical industry, and for variations inherent in food or pharmaceutical products, such as differences in content due to manufacturing changes and / or product degradation over time. This term allows for any variations in the actual food or drug to be considered in relation to the biological characteristics of the evaluated product in a subject, equivaling to the enumerated advantages of the claimed product.

[0044] The term "composition" refers to any type of composition in which multiple specific ingredients may be selectively incorporated, along with any further ingredients.

[0045] The term "include" and similar semantics should be interpreted in an open and encompassing manner as "including but not limiting".

[0046] The terms "an embodiment," "one embodiment," "a particular embodiment," etc., indicate a specific feature, attribute, or characteristic, or a group of specific features, attributes, or characteristics combined with individual semantics, presented in at least one embodiment of the invention. Such semantics appearing throughout this specification do not necessarily refer to the same embodiment. Furthermore, these specific features, attributes, or characteristics may be combined in any suitable manner in one or more embodiments.

[0047] The term "prevention" refers to the prevention of disease, condition, or symptoms, as well as the prevention of recurrence or further growth and spread of disease, condition, or symptoms after initial improvement or after initial exclusion of the cause, condition, or symptoms.

[0048] The term "treatment" refers to a therapeutic intervention that can effectively treat a disease, condition, or symptom; however, treatment can also refer to improvement, enhancement, control, control of progression, prevention of progression, and prevention of recurrence.

[0049] The term "probiotics" refers to microorganisms that contain live microorganisms, that is, microorganisms that can reproduce. Probiotics were recognized as microorganisms that, when administered in appropriate amounts, would confer health benefits to the host at the time the patent was filed.

[0050] The term "probiotic cocktail medical food composition" refers to a complex medical food containing at least one probiotic and nutrients with specific physiological functions (such as essential amino acids). This composition possesses multiple biological activities and can delay the progression of Parkinson's disease through various mechanisms (such as neuroprotection, anti-inflammation, anti-oxidation, mitochondrial activation, and gut microbiota regulation). The probiotic cocktail composition described in this invention is a complex medical nutritional formula based on a multi-component synergistic mechanism, suitable for adjunctive intervention and daily nutritional supplementation in neurodegenerative diseases.

[0051] The term "probiotics with neuroprotective or metabolic regulatory functions" refers to strains of microorganisms that can protect or stabilize the nervous system by directly or indirectly regulating neuroinflammation, antioxidation, mitochondrial function, or gut microbiota. These probiotics can be live bacteria, dead bacteria, lysate, bacterial supernatant, extracellular polysaccharides, or components with immunomodulatory functions.

[0052] The term "essential amino acids" refers to α-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 general nutritional definitions, common essential amino acids include leucine, isoleucine, valine, lysine, methionine, phenylalanine, tryptophan, threonine, and histidine.

[0053] The term "delaying the progression of Parkinson's disease" refers to slowing down or inhibiting the progression of Parkinson's disease through some kind of intervention, including but not limited to improving motor dysfunction, reducing non-motor symptoms, maintaining neuronal integrity, or improving quality of life.

[0054] The term "nonmotor symptoms associated with Parkinson's disease" includes, but is not limited to, cognitive impairment, memory impairment, disorientation, mood swings, cognitive decline, executive dysfunction, irritability, changes in interpersonal behavior, anxiety, mood swings, depression, apathy, sleep disorders, autonomic nervous system abnormalities (such as constipation, low blood pressure, urinary disorders, and abnormal sweating), and decreased sense of smell, abnormal pain perception, and fatigue.

[0055] The term "Parkinson's disease-related motor symptoms" includes, but is not limited to, tremor, muscle rigidity, bradykinesia, gait instability, dragging feet, involuntary movement disorders (such as dyskinesia, dystonia), facial expression disorders (such as mask-like face, facial paralysis), and ocular dysfunction (such as inability to open or close eyelids, blepharospasm, reduced tear film secretion, and evaporative dry eye caused by non-blinking reflex).

[0056] The term "multi-target" refers to an intervention strategy that simultaneously targets two or more physiological or molecular mechanisms related to disease pathology, such as simultaneously regulating neuroinflammatory responses, mitochondrial function, antioxidant capacity, neurotransmission regulation, or gut microbiota balance. Compared to single-target interventions, multi-target strategies can improve the overall effectiveness of interventions and contribute to the multi-level regulation of disease progression.

[0057] The *Lactobacillus reuteri* Y7 strain used in this embodiment is deposited at the Bioresource Conservation and Research Center (BCRC) in Taiwan, with accession number BCRC911162. This strain is also deposited at the International Patent Collection Center (IPOD, NITE) in Japan (Room 120, 2-5-8 Kamisamaashi, Kisarazu City, Chiba Prefecture, Japan 292-0818, Japan), with accession number NITE BP-04078.

[0058] This invention relates to a complex nutritional composition designed based on the concept of "cocktail therapy." It integrates probiotics with neuroprotective or metabolic regulatory functions with functional amino acids. Through the synergistic and complementary effects of multiple components, it regulates multiple physiological and pathological mechanisms associated with the progression of Parkinson's disease. The composition may contain probiotics from one or more sources, in the form of live bacteria, dead bacteria, processed bacterial cells, or their derivatives. It may also be combined with nutrients with potential to regulate nerve conduction, energy metabolism, and anti-inflammation, such as essential amino acids, and formulated into palatable dosage forms such as powders, tablets, capsules, and beverages in specific proportions. This composition can be used as a pharmaceutical composition, nutritional supplement, or health food, suitable for 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 dysfunction, cognitive decline, neuroinflammation and mitochondrial dysfunction, and it may regulate gut microbiota and central nervous system health through the gut-brain axis mechanism, making it an innovative nutritional intervention program with clinical application potential.

[0059] In one embodiment, the probiotics include, but are not limited to, any species of Lactobacillus, Bifidobacterium, Streptococcus, Lactococcus, Pediococcus, and Enterococcus, or a mixture thereof. The bacterial cells include, but are not limited to, live bacteria, dead bacteria, bacteria sterilized at low temperatures, lysis products, extracellular polysaccharides, or bacterial supernatant. All of the above-mentioned probiotics may affect nerve function; the Lactobacillus reuteri Y7 strain used in this embodiment is a preferred embodiment.

[0060] In one embodiment, the essential amino acid is selected from leucine, isoleucine, valine, lysine, methionine, phenylalanine, tryptophan, threonine, and histidine, and can be used alone or in combination according to different functional requirements.

[0061] In one embodiment, the improved motor symptoms may include tremor, rigidity, bradykinesia, gait abnormalities, blank facial expression, eyelid dysfunction, and involuntary movement abnormalities; while the non-motor symptoms may include anxiety, depression, sleep disorders, abnormal gastrointestinal motility, and dry eyes.

[0062] In one embodiment, the composition can be formulated into a pharmaceutical composition, nutritional supplement or health food, and prepared into powder, tablet, capsule or beverage form with appropriate pharmaceutically acceptable carriers, excipients or diluents to facilitate clinical application and long-term intake.

[0063] Example 1: The Effect of a Probiotic Cocktail Medical Food Composition on the Improvement of Parkinson's Disease Model in Rats This example aims to investigate the effect of a probiotic cocktail medical food composition containing a specific probiotic (Lactobacilus reuteriY7) and a functional amino acid combination (hereinafter referred to as "TMU-01 formulation") on the progression of Parkinson's disease. The experiment used a classic PD model preparation method to establish a 6-OHDA-induced rat Parkinson's disease model, such as... Figure 1 As shown, unilateral injection of the neurotoxin 6-hydroxydopamine (6-OHDA) into the medial forebrain bundle (MFB) of rats induced damage to dopaminergic neurons.

[0064] I. Animal Grouping and Treatment

[0065] Eighteen Sprague-Dawley rats were used in the experiment and randomly divided into three groups (n=6 in each group):

[0066] •ND group: normal control group, which did not receive 6-OHDA injection and received physiological saline by gavage;

[0067] ●PD group: Parkinson's disease model group, the right side of the MFB was injected with 6-OHDA and then received normal saline by gavage;

[0068] ●M group: The treatment group, the model was established in the same way as the PD group, and the TMU-01 formula was administered orally by gavage daily.

[0069] The experimental schedule is as follows (e.g.) Figure 1 As shown):

[0070] ● 6-OHDA injection was performed in week 0;

[0071] ● Apomorphine-induced rotational tests were performed in weeks 2 and 10;

[0072] • The Rotarod test, grip strength test, and Morris water maze test will be conducted in weeks 8 and 9.

[0073] ● Animals were sacrificed in week 10 for tissue staining and subsequent analysis.

[0074] II. Behavioral Function Assessment

[0075] Apomorphine-induced rotational behavior tests were conducted at weeks 2 and 10. Results showed that the PD group had significantly more rotations than the ND group in both tests (e.g., ...). Figure 2 China A and Figure 2 As shown in Figure B), the motor function of the group was significantly impaired; while the number of rotations in the M group was significantly lower than that in the PD group, indicating that this formula can effectively improve abnormal rotational behavior.

[0076] In the Rotarod balance test, the PD group rats had a significantly shorter time on the platform, indicating a decline in their motor coordination function; the M group, on the other hand, performed significantly better than the PD group in this indicator (e.g., Figure 2 As shown in Figure C, this indicates that its motion stability and balance have recovered to some extent.

[0077] Overall behavioral assessment results showed that this formula could significantly improve motor dysfunction in a 6-OHDA-induced Parkinson's disease rat model, including reducing rotational behavior and improving motor coordination and muscle strength, demonstrating its potential neuroprotective and motor function recovery benefits.

[0078] III. Cognitive Function Assessment

[0079] To assess the effect of this formula on cognitive function, the Morris water maze test was conducted in weeks 8 to 9 to evaluate spatial learning and memory abilities.

[0080] like Figure 3 China A to Figure 3 As shown in Figure C, the PD group rats exhibited a significantly prolonged escape latency, a significantly reduced time spent in the target quadrant, and an increased total swimming distance, indicating that their spatial learning ability and memory recall ability were significantly impaired.

[0081] In comparison, the M group rats showed significantly better performance than the PD group in terms of escape latency and time spent in the target area. This formula can effectively improve spatial learning and memory abilities caused by Parkinson's disease, and its cognitive performance is close to that of the ND group, indicating that this formula can improve cognitive impairment caused by Parkinson's disease.

[0082] Overall, the Morris water maze results showed that this formula has a significant protective effect on spatial learning and memory abilities in 6-OHDA-induced PD models, and helps to delay Parkinson's disease-related cognitive decline.

[0083] IV. Neuroprotective Effect

[0084] 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, and brain samples were collected for tyrosine hydroxylase (TH) immunohistochemical staining to analyze the distribution and number of TH-positive neurons in the substantia nigra pars compacta (SNpc).

[0085] like Figure 4 China A and Figure 4 As shown in Figure B, the number of TH-positive cells in the SNpc region of the PD group rats was significantly lower than that in the ND group, indicating the loss of dopaminergic neurons, which is consistent with the neuropathological characteristics of Parkinson's disease.

[0086] The number of TH-positive cells in the M group rats was significantly preserved, and the number of positive cells was significantly higher than that in the PD group. This indicates that the intervention of this formula can effectively inhibit neuronal damage and loss, showing a dopamine neuroprotective effect.

[0087] These results demonstrate that, in addition to improving behavioral and cognitive performance, this formula can also significantly preserve dopamine neurons at the tissue level, with a significant increase in the number of TH-positive cells, indicating that this formula has a neuroprotective effect and the potential to delay the pathological progression of Parkinson's disease.

[0088] V. Analysis of Inflammation and Antioxidant Indicators

[0089] To further investigate the regulatory effects of this formulation on neuroinflammation and oxidative stress, striatal tissue was collected from rats at week 10 for mRNA expression analysis of pro-inflammatory and antioxidant genes.

[0090] like Figure 5 China A and Figure 5As shown in Figure B, compared with the normal control ND group, the PD group rats showed significantly increased levels of inflammatory factors tumor necrosis factor-α (Tnf-α) and interleukin-6 (Il-6), indicating an increased neuroinflammatory response in the Parkinson's disease model. In contrast, the M group rats showed significantly lower levels of inflammatory factors Tnf-α and Il-6 than the PD group, indicating that this formula has a significant anti-inflammatory effect.

[0091] like Figure 5 C and Figure 5 As shown in Figure D, the expression levels of the antioxidant enzymes glutathione peroxidase (Gpx) and catalase (Cat) in the PD group were significantly lower than those in the ND group, indicating that their antioxidant system was impaired. The expression of Gpx and Cat genes in the M group rats was significantly higher than that in the PD group, indicating that this formula can enhance the antioxidant capacity of cells and strengthen the defense mechanisms of nerve cells against oxidative stress.

[0092] In summary, this formula can simultaneously reduce the expression of neuroinflammatory factors and enhance the activity of antioxidant enzyme genes, thus possessing the dual benefits of regulating neuroinflammation and oxidative stress, which helps protect nerves and delay neurodegeneration.

[0093] VI. Mitochondrial Functional Analysis

[0094] To assess the effect of this formulation on mitochondrial function, striatum and soleus muscle tissues from rats were collected for analysis of metabolic function-related indicators, including oxygen consumption rate (OCR), extracellular acidification rate (ECAR), and expression levels of mitochondrial biosynthesis-related genes Sirtuin 1 (Sirt1) and Peroxisome proliferator-activated receptor gammacoactivator 1-alpha (Pgc-1α).

[0095] like Figure 6 China A and Figure 6 As shown in Figure B, the OCR and ECAR in the striatum of rats in the PD group were significantly lower than those in the ND group, indicating mitochondrial dysfunction. Mitochondrial function in the M group was significantly restored, with both OCR and ECAR being significantly higher than those in the PD group, indicating a significant improvement in mitochondrial function.

[0096] Further analysis of genes related to mitochondrial synthesis revealed a significant 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 (e.g., ...). Figure 6 C and Figure 6 As shown in Figure D, this further supports the recovery and enhancement of mitochondrial function.

[0097] Furthermore, similar results have been observed in muscle tissue, such as... Figure 7 China A to Figure 7 As shown in Figure D, in the soleus muscle, the OCR, ECAR, Sirt1, and Pgc-1α levels in the PD group decreased, while those in the M group significantly increased. This indicates that the formula not only acts on the central nervous system but also has a repairing effect on the mitochondrial function of peripheral tissues. The formula can improve mitochondrial health systemically.

[0098] In summary, this formula can effectively improve neurological dysfunction caused by energy disorders in animal models of Parkinson's disease by restoring mitochondrial energy metabolism and enhancing mitochondrial production, and has the potential for systemic mitochondrial regulation.

[0099] VII. Intestinal flora analysis

[0100] To evaluate the regulatory effect of this formulation on the composition of the gut microbiota, rat fecal samples were collected at week 10 of the experiment for 16S rRNA high-throughput sequencing analysis, and the results were compared with the LEfSe statistical method by relative abundance analysis of the microbiota.

[0101] like Figure 8 China A to Figure 8 As shown in Figure C, the gut microbiota structure of PD group rats was significantly altered at the phylum, genus, and species levels. The relative abundance of probiotics decreased, and an increase in various potential pathogens was observed, indicating that the PD model leads to gut microbiota imbalance.

[0102] The intestinal flora structure of rats in group M was significantly different from that in group PD, and it had the effect of restoring the relative abundance of beneficial bacteria and improving microbial diversity. Its flora distribution was close to that of group ND, indicating that this formula can reconstruct the damaged intestinal microecology in the Parkinson's disease model.

[0103] In addition, such as Figure 9 China A and Figure 9 As shown in Figure B, LEfSe analysis compared the differences in gut microbiota among the three groups and found that group M had gut microbiota characteristics that were clearly distinguishable from group PD. This indicates that the intervention of this formula can lead to an increase in the dominance of specific beneficial bacteria, showing that it has a good regulatory effect on the composition of gut microbiota and can restore gut microbial diversity and ecological balance.

[0104] The results show that the TMU-01 formula has a good gut microbiota regulation function, which can restore gut microbial diversity and ecological balance. It may indirectly alleviate the neurodegenerative symptoms associated with Parkinson's disease through the gut-brain axis mechanism, providing potential systemic disease regulation benefits.

[0105] The results of the embodiments of the present invention show that the TMU-01 formulation (a cocktail medical food composition containing probiotics and a combination of functional amino acids) can exhibit multiple significant physiological improvement effects in a 6-OHDA-induced Parkinson's disease animal model, and has the following technical effects:

[0106] 1. Improvement of motor dysfunction: Effectively reduces apomorphine-induced rotational behavior, improves Rotarod platform dwell time and grip strength test performance, showing that it can restore motor function in PD model rats (e.g., Figure 2 (As shown).

[0107] 2. Enhanced cognitive function: Improved spatial learning and memory performance in the Morris water maze, shortened escape latency, increased time spent in the target area, demonstrating a protective effect of spatial memory (e.g., Figure 3 (As shown).

[0108] 3. Protection of dopamine neurons: The number of TH-positive cells in the SNpc region was significantly increased, indicating a neuroprotective effect and delaying the degeneration of dopamine neurons (e.g., Figure 4 (As shown).

[0109] 4. Enhanced anti-inflammatory and antioxidant capabilities: Reduces the expression of inflammatory factors Tnf-α and Il-6, and enhances the expression of antioxidant-related genes Gpx and Cat, helping to stabilize the neural microenvironment (e.g., Figure 5 (As shown).

[0110] 5. Restoring mitochondrial metabolic function: It increases OCR and ECAR values ​​in the striatum and soleus muscle, and promotes the expression of mitochondrial biosynthesis genes Sirt1 and Pgc-1α, demonstrating a restorative effect on mitochondrial function and enhanced energy metabolism (e.g., Figure 6 and Figure 7 (As shown).

[0111] 6. Regulating gut microbiota and restoring microecological balance: Significantly increases the relative abundance of probiotics, improves the gut microbiota imbalance associated with Parkinson's disease, and confirms significant differences in microbiota indicators (such as...) through LEfSe analysis. Figure 8 and Figure 9 (As shown).

[0112] The results above demonstrate that this formula possesses multiple regulatory mechanisms, acting simultaneously on the central nervous system and peripheral tissues. It exhibits significant improvements in behavioral function, neuropathology, molecular markers, and systemic metabolism in animal models of Parkinson's disease. This formula holds promise as an adjunctive nutritional therapy with the potential to delay disease progression, and can be applied to preventative healthcare, early intervention, or disease management for Parkinson's disease, possessing high clinical and industrial application value.

[0113] The innovation of this invention lies in providing a medical food composition combining probiotics and functional amino acids to specifically regulate key aspects related to the progression of Parkinson's disease, such as neurodegeneration, inflammatory responses, mitochondrial dysfunction, and cognitive decline. This combination exhibits multiple mechanisms of action and complementary functions, significantly outperforming existing products containing only single ingredients, thus delaying the pathological progression of PD from its source. This invention uses Parkinson's disease as a specific indication and employs a 6-OHDA-induced Parkinson's disease animal model for systematic validation, covering multiple levels of indicators including motor behavior, cognitive performance, dopaminergic neuron integrity, inflammatory and antioxidant markers, mitochondrial function, and gut microbiota structure. Experimental results show that this composition can significantly improve motor and memory function in Parkinson's disease model animals, protect dopaminergic neurons, reduce neuroinflammation, enhance antioxidant capacity, restore mitochondrial metabolic activity in brain and muscle tissue, and rebuild a gut microbiota structure dominated by beneficial bacteria. The aforementioned efficacy is closely related to the progression of Parkinson's disease, indicating that this formulation represents a non-pharmacological nutritional intervention strategy with disease-modifying potential. Compared to traditional drugs that only alleviate symptoms, the medical food composition of this invention can serve as a long-term supportive disease-modifying intervention, suitable for daily nutritional supplementation for early-stage Parkinson's disease patients or high-risk groups. Furthermore, this composition can be formulated as powder, tablets, capsules, or beverages for convenient long-term intake and clinical implementation. Besides serving as an adjunctive treatment option for Parkinson's disease patients, it is also suitable for preventative nutritional fortification in the general population, possessing high clinical translational value and industrial application potential.

[0114] Biomaterial storage

[0115] The strain was deposited at the Food Industry Development Research Institute Foundation on November 29, 2022, with deposit number BCRC911162. It was also deposited at the National Institute of Advanced Industrial Science and Technology, Japan, Patent Biological Collection Center (IPOD, NITE) (Room 2-5-8120, Kazusa-Kamazutari, Kisarazu City, Chiba Prefecture 292-0818, Japan), on February 9, 2024, with deposit number NITE BP-04078.

Claims

1. A composition, characterized in that, The product comprises (a) at least one probiotic with neuroprotective or metabolic regulatory functions; and (b) a functional amino acid combination comprising four or more essential amino acids.

2. The composition according to claim 1, characterized in that, The probiotic strains with neuroprotective or metabolic regulatory functions are selected from a group consisting of Lactobacillus, Lactococcus, Streptococcus, Bifidobacterium, Pediococcus, and Enterococcus.

3. The composition according to claim 2, characterized in that, The Lactobacillus species mentioned are selected from 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. The group consisting of .rhamnosus, L.sakei, L.salivarius, L.paracasei, L.kisonensis, L.paralimentarius, L.perolens, L.apis, L.ghanensis, L.dextrinicus, L.shenzenensis and L.harbinensis.

4. The composition according to claim 1, characterized in that, The probiotics are live bacteria, dead bacteria, bacteria that have undergone low-temperature sterilization, lysis products, bacterial supernatant, cell wall extracts, extracellular polysaccharides, or their immunostimulatory components.

5. The composition according to claim 1, characterized in that, The essential amino acid is selected from leucine, isoleucine, valine, lysine, methionine, phenylalanine, tryptophan, threonine, and histidine.

6. The composition according to claim 1, characterized in that, The composition is a pharmaceutical composition, nutritional supplement, or health food.

7. The composition according to claim 1, characterized in that, The composition includes pharmaceutically acceptable carriers, excipients, and diluents.

8. The composition according to claim 1, characterized in that, The composition is in the form of powder, tablets, capsules, or beverage.

9. Use of the composition of claim 1 in the preparation of a medicament for delaying the progression of Parkinson's disease.

10. The use according to claim 9, characterized in that, The delay in the progression of Parkinson's disease refers to the reduction of Parkinson's disease-related motor symptoms and / or the reduction of Parkinson's disease-related non-motor symptoms.

11. The use according to claim 10, characterized in that, The relief of nonmotor symptoms associated with Parkinson's disease includes cognitive impairment, memory impairment, disorientation, cognitive decline, executive dysfunction, irritability, changes in interpersonal behavior, anxiety, mood swings, depression, apathy, sleep disorders, autonomic dysfunction, olfactory dysfunction, abnormal pain perception, or fatigue.

12. The use according to claim 11, characterized in that, The autonomic nervous system dysfunction includes constipation, low blood pressure, urinary disorders, or abnormal sweating.

13. The use according to claim 12, characterized in that, The reduction of Parkinson's disease-related motor symptoms includes tremor, muscle rigidity, bradykinesia, gait instability, foot dragging, involuntary movement disorders, facial expression disorders, and eye dysfunction.

14. The use according to claim 13, characterized in that, The involuntary movement abnormalities include difficulty moving and muscle dysregulation.

15. The use according to claim 13, characterized in that, The facial expression disorder includes mask face and facial paralysis.

16. The use according to claim 13, characterized in that, The ocular dysfunctions include inability to open and close the eyelids, blepharospasm, reduced tear film secretion, and evaporative dry eye caused by non-blinking response.

17. The use according to claim 9, characterized in that, Delaying the progression of Parkinson's disease can be achieved by enhancing anti-inflammatory capabilities, boosting antioxidant capacity, restoring mitochondrial metabolic function, adjusting gut microbiota, or rebuilding gut microecological balance.

18. The use according to claim 10, characterized in that, The composition is intended for daily nutritional supplementation for patients with Parkinson's disease or at high risk of developing Parkinson's disease to delay disease progression.

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