A medical food formulation supplementing with a probiotic cocktail can extend the stability of L-DOPA administration in Parkinson's disease patients and reduce the occurrence of side effects from long-term administration.

Abstract

This formulation can be applied to Parkinson's disease patients to improve the stability of levodopa (L-DOPA) in the body and its availability to the central nervous system. This formula can regulate the gut microbiota, strengthen the intestinal barrier function, reduce systemic inflammation and oxidative pressure, further improve motor and non-motor dysfunction, and delay or reduce the occurrence of side effects of long-term L-DOPA treatment, such as dysregulation. [Solution] The present invention discloses a probiotic cocktail medical food formulation that comprises at least one probiotic and can selectively combine one or more amino acids.

Description

Technical Field

[0001] The present invention relates to a probiotic catheter medical food formulation having a medical assistance function, and particularly to a medical food formulation for improving treatment stability and side effects associated with long-term use of L-DOPA by Parkinson's disease patients. This formulation contains at least one probiotic and selectively contains amino acid components, which can regulate the intestinal flora, improve the metabolic stability of L-DOPA, further enhance the effect, and reduce side effects.

Background Art

[0002] Parkinson's disease (PD) is a chronic neurodegenerative disease characterized by the decline of central dopamine neurons and is commonly seen in middle-aged and elderly people. Its main clinical symptoms include movement disorders (such as tremors, rigidity, slow movements) and non-motor symptoms (such as constipation, cognitive impairment, sleep disorders, etc.).

[0003] Currently, the forefront drug for the clinical treatment of Parkinson's disease is levodopa (L-DOPA). As a precursor of dopamine, it can effectively supplement the content of dopamine in the brain and can significantly improve the patient's movement symptoms in the short term. However, L-DOPA has obvious treatment limitations: (1) Long-term use is likely to cause fluctuations in the treatment effect (on-off phenomenon), (2) It can cause abnormal involuntary movements (L-DOPA-induced dyskinesia, LID), (3) L-DOPA has a fast metabolism in the periphery, and the effective concentration actually entering the central nervous system decreases, (4) Long-term use may also worsen non-motor symptoms.

[0004] In addition to L-DOPA, current treatment methods also include the use of drugs such as COMT inhibitors, MAO-B inhibitors, and dopamine receptor agonists, but there are also problems of side effects and drug resistance, and the stability of L-DOPA and the risk of side effects cannot be effectively solved.

[0005] Recent studies suggest that the gut microbiota can be involved in the metabolism and absorption of L-DOPA, and certain bacterial species (e.g., Clostridium and Enterococcus) can break down L-DOPA early in the gut, reducing its efficiency in entering the bloodstream and brain. Furthermore, intestinal barrier dysfunction and chronic inflammation exacerbate the progression of Parkinson's disease and the side effects of treatment.

[0006] Therefore, regulating the intestinal environment and improving the bacterial flora composition and barrier integrity is expected to be an innovative approach to enhancing the stability and safety of L-DOPA. With current technology, the design of comprehensive supportive therapies for this strategy is still insufficient. In particular, there is a lack of innovative supportive intervention strategies that are safe, can be used long-term, and have intestinal regulatory functions, especially in terms of enhancing the therapeutic effect of L-DOPA, stabilizing its drug metabolism, and mitigating side effects. There is an urgent need to develop supportive therapies that are suitable for long-term use, highly safe, and have a comprehensive effect. [Overview of the project] [Problems that the invention aims to solve]

[0007] The primary objective of the present invention is to provide a probiotic cocktail medical food formulation that can slow the peripheral metabolic rate of L-DOPA in the body of Parkinson's disease patients, thereby increasing its stability and availability to the central nervous system, enhancing therapeutic effects, and reducing side effects associated with long-term treatment. The formulation of the present invention can also improve the stability and integrity of the intestinal bacterial community and further modulate inflammatory responses and neurological function. [Means for solving the problem]

[0008] To achieve the above objective, the present invention proposes an application for producing a composition that alleviates or improves the symptoms of Parkinson's disease, wherein the composition comprises at least one probiotic having intestinal regulatory or neuroprotective function.

[0009] To achieve the aforementioned inventive objective, the composition further comprises a combination of functional amino acids, which includes four or more essential amino acids.

[0010] To achieve the aforementioned inventive objectives, this application further includes extending the stability of administration of drugs effective for Parkinson's disease and reducing side effects of administration.

[0011] To achieve the aforementioned inventive objectives, extending the stability of administration of this Parkinson's disease-effective drug and reducing its side effects would involve increasing the central availability of levodopa (L-DOPA), delaying abnormal involuntary movements (L-DOPA-induced dyskinesia, LID), improving non-motor symptoms, reducing inflammatory and oxidative stress responses, strengthening the integrity of the intestinal barrier, or improving the balance of the intestinal bacterial community, non-motor symptoms, and cognitive symptoms.

[0012] To achieve the aforementioned objective of the invention, the drug effective for Parkinson's disease is selected from the group consisting of levodopa (L-DOPA), dopamine agonists, MAO-B inhibitors, DOPA decarboxylase inhibitors, and COMT inhibitors.

[0013] To achieve the aforementioned inventive objectives, the probiotics are selected from the group consisting of the genera Lactobacillus, Lactococcus, Streptococcus, Bifidobacterium, Pediococcus, and Enterococcus.

[0014] To achieve the aforementioned inventive objective, the Lactobacillus genus is selected here from the group consisting of L. acidophilus, L. antri, L. brevis, L. casei, L. coleohominis, L. crispatus, L. curvatus, L. fermentum, L. gasserri, 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.

[0015] To achieve the aforementioned inventive objectives, the probiotics here include live bacteria, dead bacteria, pasteurized bacteria, degradation products, bacterial supernatant, cell wall extracts, extracellular polysaccharides, or their immunostimulant components.

[0016] To achieve the aforementioned inventive objective, the drug effective in treating Parkinson's disease is levodopa (L-DOPA).

[0017] To achieve the aforementioned inventive objectives, the essential amino acids are selected from leucine, isoleucine, valine, lysine, methionine, phenylalanine, tryptophan, threonine, and histidine.

[0018] To achieve the aforementioned objectives of the invention, the non-motor symptoms here include cognitive impairment, gastrointestinal dysfunction, or sleep disorders.

[0019] To achieve the aforementioned inventive objectives, this composition improves Parkinson's disease behavioral function and delays L-DOPA-induced abnormal involuntary movements through overall gut-brain axis regulation and neuroprotective effects.

[0020] To achieve the aforementioned objectives of the invention, this composition hereby improves motor and non-motor symptoms of Parkinson's disease and reduces L-DOPA side effects by regulating the intestinal bacterial flora, reducing systemic inflammation, and improving the central availability of L-DOPA.

[0021] To achieve the aforementioned inventive objective, here, Parkinson's disease refers to a person in the prodromal, early, or late stages of Parkinson's disease, or a person who has not received treatment.

[0022] Another object of the present invention is to provide a composition for improving Parkinson's disease symptoms, extending the stability of administration of drugs effective for Parkinson's disease, and reducing side effects of administration, comprising (a) at least one probiotic bacterium having intestinal regulatory or neuroprotective function, and (2) a combination of functional amino acids comprising four or more essential amino acids.

[0023] To achieve the aforementioned objective of the invention, the drug effective in treating Parkinson's disease is selected from the group consisting of levodopa (L-DOPA), dopamine agonists, MAO-B inhibitors, DOPA decarboxylase inhibitors, and COMT inhibitors.

[0024] To achieve the aforementioned inventive objective, the drug effective in treating Parkinson's disease is levodopa (L-DOPA).

[0025] To achieve the aforementioned inventive objectives, the probiotic is selected from the genera Lactobacillus, Bifidobacterium, or Streptococcus.

[0026] To achieve the above-described object of the invention, here, this essential amino acid is selected from leucine, isoleucine, valine, lysine, methionine, phenylalanine, tryptophan, threonine, and histidine.

[0027] To achieve the above-described object of the invention, here, this branched-chain amino acid is selected from silamic acid, isosilamic acid, and valinic acid.

[0028] Another object of the present invention is to provide a pharmaceutical composition for improving Parkinson's disease symptoms, which comprises (a) a drug effective for treating Parkinson's disease, (b) at least one probiotic bacterium having a tumor agent intestinal regulation or neuroprotective function, and (c) a combination of functional amino acids containing four or more essential amino acids.

[0029] To achieve the above-described object of the invention, here, this drug effective for treating Parkinson's disease is L-DOPA.

[0030] To achieve the above-described object of the invention, here, this probiotic is L. reuteri.

[0031] To achieve the above-described object of the invention, here, this essential amino acid is selected from leucine, isoleucine, valine, lysine, methionine, phenylalanine, tryptophan, threonine, and histidine.

[0032] To achieve the above-described object of the invention, this pharmaceutical composition can extend the stability of the administration of a drug effective for Parkinson's disease and reduce the administration side effects. [Effects of the Invention]

[0033] As described above, the present invention provides a probiotic cocktail medical food formulation, a composition or pharmaceutical composition mainly consisting of a combination of probiotics and functional amino acids, which can effectively improve motor and non-motor symptoms in Parkinson's disease (PD) patients through the regulation of the gut-brain axis and neuroprotective mechanisms, and can significantly improve the stability and utilization of therapeutic drugs such as levodopa (L-DOPA) in the central nervous system. This composition can alleviate abnormal involuntary movement (LID) side effects associated with L-DOPA, and can further delay the progression of the disease by reducing inflammation and oxidative pressure, strengthening the intestinal barrier function, and regulating the intestinal bacterial flora. Compared to conventional single-pharmacological therapies, the present invention offers the advantage of regulating multiple physiological functions, is safe in composition, is applicable long-term, substantially contributes to the effectiveness of Parkinson's disease treatment and improves the quality of life for patients, and has high potential for industrial application. [Brief explanation of the drawing]

[0034] [Figure 1] Explanation of the timeline for experimental animals. The overall flow of 6-OHDA induction, prescribed administration, and behavioral testing (Example 1) is shown. [Figure 2] Immunohistochemical evaluation of dopaminergic neurons in the striatum and substantia nigra pars compacta (SNpc) after 8 weeks of treatment. (A) TH immunostaining images, (B) Quantitative results of TH-positive neurons. One-way ANOVA and Tukey's post hoc test were used for statistical analysis. #p<0.05 indicates a significant difference between the PD group and the NC group, and *p<0.05 indicates a significant difference between the Y group and the PD group. [Figure 3] PD rat motor function tests. (A) Rotarod behavior test, (B) grip strength test, (C) gait test. Data are expressed as mean ± SD, and the statistical method is the same as in Figure 2. [Figure 4]PD rat non-motor function tests. (A) Fecal volume, (B) Fecal water content, (C) Expression in Morris water maze, (DF) Analysis of intestinal adhesion gene expression levels of zo1, occ, and claudin. [Figure 5] Results of (A) expression of inflammatory cell hormones IL-6, IL-1β, and TNF-α, and (B) expression of oxidative pressure-related markers (ROS, SOD, and GPx) in rat serum from each group. [Figure 6] Relative abundance and diversity analysis of the gut microbiota after 8 weeks of treatment. (A) Genus-level microbiota distribution, (B) α-diversity analysis (Shannon and Simpson indices), (C) Principal coordinate analysis (PCoA) diagram. [Figure 7] Bacterial species showing significant differences between groups based on LEfSe and metaSeq analysis identification. (A) Heatmap based on LEfSe analysis, (B) Clear bacterial flora histogram from metaSeq analysis. [Figure 8] Explanation of the time axis of experimental animals (Example 2-1). [Figure 9] TH immunohistochemical analysis of dopaminergic neurons in the substantia nigra pars compacta (SNpc) after 8 weeks of treatment. (A) TH immunostaining diagram, (B) Quantitative results of TH-positive cells. [Figure 10] Motor function test results of PD rats. (AB) Rotarod expression, (C) Grip strength test. [Figure 11] Analysis of L-DOPA utilization rates in different treatment groups. (AC) Serum L-DOPA concentration, (DF) Serum dopamine concentration, (G) Ratio of L-DOPA to DA concentration. [Figure 12] Colon inflammation-related gene expression analysis. (A) TLR4, (B) NF-κB, (C) TNF-α, (D) IL-6. [Figure 13] Colon-adherent gene expression analysis. (A)ZO-1, (B)Occludin, (C)Claudin-1. [Figure 14] Explanation of the time axis for experimental animals (Example 2-2). [Figure 15] AIMs (Ambient Movement Instincts) behavioral scoring method and time axis. [Figure 16] Aiminess Independence Score (AIM score). [Figure 17] Cognitive function analysis of the Morris water maze test. (A) Trajectory diagram of the test representative, (B) Quantification of test duration, (C) Quantification of total distance traveled. [Figure 18] L-DOPA stability and metabolic analysis. (A) L-DOPA concentration, (B) DA concentration, (C) L-DOPA to DA ratio. [Figure 19] Relative abundance of enteric bacterial flora in each group. (A) Firmicutes / Bacteroidota ratio, (B) door-level, (C) genus-level distribution. [Figure 20] Bacterial diversity analysis. (A) α-diversity analysis (Shannon and Simpson), (B) PCoA principal coordinate analysis. [Figure 21] LEfSe analysis results. (A) Bar graph of LDA scores for bacterial communities with significant differences, (B) Heatmap of bacterial communities from different groups. [Modes for carrying out the invention]

[0035] 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.

[0036] The terms “one” or “one kind” do not exclude multiple things. That is, unless the meaning is clearly expressed or required, a single type of “one,” “one kind,” or “this” should be understood to include multiple subjects. In other words, unless specifically expressed or unless the meaning of the cited term 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. Therefore, unless specifically defined, the terms “one” and “one kind” are synonymous with “this” and “at least one” or “one or more.” For example, when referring to “one component,” this includes mixtures of multiple components, etc.

[0037] The terms "approximately" or "about" are acceptable in the pharmaceutical industry to compensate for variations in drug content, such as differences in content due to manufacturing variations and / or product degradation over time. Allowing any actual drug modification with these terms should be done considering that the bioproperties of the product being evaluated in subjects are equivalent to the enumerated merits requiring protection of the product.

[0038] The term "composition" means any type of composition, and several specific components are selectively included in this composition along with any further components.

[0039] The term "includes" should be interpreted in an open and inclusive way, meaning "includes, but does not limit."

[0040] The terms "one embodiment," "a single embodiment," and "a specific embodiment" mean that one particular feature, attribute, or characteristic, or a group of particular features, attributes, or characteristics, is included in at least one embodiment of the present invention in combination with the individual vocabulary. Such meanings appearing throughout this patent specification do not necessarily mean the same embodiment. Furthermore, these particular features, attributes, or characteristics may be combined in any suitable manner in one or more embodiments.

[0041] The term "medically acceptable" means a compound or mixture for the manufacture of a pharmaceutical composition that is generally safe, non-toxic, and not biologically or otherwise undesirable, and this includes compounds or mixtures suitable for medicinal use in the human body.

[0042] The term "prevention" means preventing a disease, condition, or symptom, and after the etiology, condition, or symptom of the disease has been improved or eliminated, further increase and spread of recurrence or progression must be prevented.

[0043] The term "treatment" refers to a therapeutic intervention that can effectively treat a disease, condition, or symptom, but it can also mean improvement, advancement, control, suppression of progression, prevention of progression, prevention of relapse, etc.

[0044] 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 at the technological level at the time of the patent application, provide health benefits to the host.

[0045] The term "probiotics with intestinal regulatory function" refers to microorganisms that promote the balance of the intestinal microbial community, improve intestinal barrier function, reduce intestinal inflammatory responses, and improve gastrointestinal function, and includes, but is not limited to, genera of Lactobacillus, Lactococcus, Streptococcus, Bifidobacterium, Pediococcus, and Enterococcus.

[0046] The term "therapeutic dose" means that when administering a drug to a subject to treat or prevent a target disease (e.g., Parkinson's disease), the dose is sufficient to achieve the expected therapeutic, palliative, preventive, or ameliorative effect, but does not necessarily mean that all symptoms will be completely resolved.

[0047] The term "neuroprotective function" refers to physiological effects that can mitigate nerve deterioration, reduce nerve cell death, or promote nerve repair and reconstruction.

[0048] The term "abnormal involuntary movements (L-DOPA-induced dyskinesia, LID)" refers to involuntary, rhythmic, or pendulum-like movements that appear in Parkinson's disease patients after long-term use of L-DOPA, and is a common but difficult-to-control side effect of the drug.

[0049] The term "gut-brain axis" refers to the bidirectional regulatory pathway between the gut microbiota, the intestines, and the central nervous system, and is involved in multiple mechanisms including nerve, immune, and metabolic processes.

[0050] The term "central nervous system availability" refers to the extent to which an active drug component enters the central nervous system (e.g., the brain), and this parameter has a significant impact on the onset of drug efficacy.

[0051] The term "intestinal barrier integrity" refers to the functional integrity of the intestinal epithelial cell layer and its tight junctions, which effectively prevents harmful substances (such as endotoxins and pathogens) from entering and circulating within the body's systems, thereby maintaining the physiological stability of the intestinal environment and immune barrier function.

[0052] The term "intestinal microbiota balance" refers to a state where the diversity and relative proportions of intestinal probiotics and other symbiotic bacteria are stable, positively impacting host health, immune regulation, and metabolic function. Microbiota imbalances may be associated with many neurodegenerative diseases, including Parkinson's disease.

[0053] The term "non-motor symptoms" refers to other systemic symptoms commonly seen in Parkinson's disease patients besides motor impairments, and includes, but is not limited to, emotional disturbances, cognitive impairments, autonomic dysfunction (e.g., gastrointestinal disorders, urinary disorders), sleep disturbances, pain, and fatigue.

[0054] The term "improvement of cognitive symptoms" refers to enhancing a patient's cognitive function, including memory, attention, executive function, and language comprehension. For Parkinson's disease patients, cognitive symptoms are a common progression of the disease and significantly impact their quality of life.

[0055] The term "essential amino acids" 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 are selected from leucine, isoleucine, valine, lysine, methionine, phenylalanine, tryptophan, threonine, and histidine.

[0056] The present invention relates to a probiotic cocktail medical food formulation for improving or alleviating symptoms of Parkinson's disease, wherein the probiotic cocktail medical food formulation comprises at least one probiotic having intestinal regulatory or neuroprotective functions, and may further comprise serotonic acid and at least one branched-chain amino acid, such as silaminic acid, isocillamic acid, and valic acid. When used as an adjunctive therapy in combination with drugs effective for Parkinson's disease, this composition improves the central nervous system availability of the drugs, extends the stability of their efficacy, and reduces associated side effects, such as abnormal involuntary movements (L-DOPA-induced dysregulation, LID), cognitive impairment, gastrointestinal dysfunction, and sleep disorders.

[0057] In one embodiment, a composition for alleviating Parkinson's disease symptoms is provided, which comprises at least one probiotic having intestinal regulatory or neuroprotective function. This probiotic can indirectly improve brain neurotransmission by regulating the intestinal bacterial community and strengthening the intestinal barrier, and can further alleviate motor or non-motor symptoms in patients with Parkinson's disease.

[0058] In one embodiment, the composition further comprises a combination of functional amino acids.

[0059] In one embodiment, this composition can be used in combination with drugs effective for Parkinson's disease to enhance the stability of their efficacy and mitigate side effects associated with long-term medication. These side effects include abnormal involuntary movements (L-DOPA-induced dyskinesia, LID), sleep disorders, gastrointestinal dysfunction, and mood disorders.

[0060] In one embodiment, the mechanism of action of this composition includes improved central availability of L-DOPA, delayed LID development, reduced inflammation and oxidative pressure, improvement of non-motor symptoms (e.g., cognitive impairment), and maintenance of the balance of the intestinal microbiota. This comprehensive mechanism of action indicates that the composition has the potential for multiple targeted adjuvant therapies.

[0061] In one embodiment, the drug effective for Parkinson's disease is selected from levodopa (L-DOPA), dopamine agonists, MAO-B inhibitors, DOPA decarboxylase inhibitors, and COMT inhibitors.

[0062] In one embodiment, the probiotics are selected from any of the following bacterial species or mixtures: Lactobacillus, Bifidobacterium, Streptococcus, Lactococcus, Pediococcus, and Enterococcus. The bacterial cells may be live, dead, pasteurized, decomposition products, extracellular polysaccharides, or bacterial supernatant.

[0063] In one embodiment, the essential amino acid is selected from leucine, isoleucine, valine, lysine, methionine, phenylalanine, tryptophan, threonine, and histidine.

[0064] In one embodiment, this composition may further provide neuroprotective effects by regulating the overall gut-brain axis, improving cognitive and motor function expression, and potentially delaying disease progression as an adjunct strategy to long-term L-DOPA treatment.

[0065] In one embodiment, the composition may be manufactured in the form of a powder, capsule, liquid, or fermented food, depending on clinical needs, and may be used as a routine health or drug adjunct prescription, or administered in a stepwise manner in combination with drugs to adjust the amount of L-DOPA used and reduce the risk of tolerance.

[0066] The Lactobacillus reuteri Y7 used in this embodiment is deposited with the Center for Biological Resource Conservation and Research (BCRC), a Taiwanese foundation, with deposit number BCRC 911162. The aforementioned strain is also deposited with the International Patent and Biodiversity Preservation Center (IPOD, NITE), a Japanese international depositary organization located at Room 120, 2-5-8 Kazusa-Kamatari, Kisarazu City, Chiba Prefecture, 292-0818 Japan, with deposit number NITE BP-04078.

[0067] Example 1: Evaluation of the effect of probiotic Y7 on improving PD motor and non-motor impairments.

[0068] This study aims to investigate whether the probiotic Y7 can improve motor and non-motor deficits in a rat model of Parkinson's disease (PD). The experiment involved inducing dopaminergic neuronal damage by unilaterally injecting the medial forebrain bundle (MFB) of rats with the neurotoxin 6-hydroxydopamine (6-OHDA), as shown in Figure 1, using a typical PD model preparation method. Eighteen male Sprague-Dawley rats were selected and randomly divided into a normal control group (NC group), a PD model group (PD group), and a probiotic Y7 intervention group (Y group). The Y group received daily oral intragastric administration of probiotic Y7 once daily for nine consecutive weeks. Behavioral tests were performed at weeks 8-9, and tissue and serum samples were collected and analyzed.

[0069] Two weeks after modeling, the rats in the PD model group clearly exhibited a behavior of turning towards the lesion on an automated rotation test device after receiving apomorphine via intraperitoneal injection, confirming the success of the model creation.

[0070] To evaluate the neuroprotective effects on dopaminergic neurons, samples were taken and subjected to tyrosine hydroxylase (TH) immunohistochemical staining, and the number of surviving dopaminergic neurons in the striatum and substantia nigra pars compacta (SNpc) was analyzed. The results showed a clear decrease in TH-positive neurons in the PD group, but a clear recovery in the Y group (Figures 2A, 2B), indicating that the probiotic Y7 has neuroprotective potential.

[0071] In terms of motor function, the Rotarod test results showed that the Y group could significantly increase the distance traveled along the axis of rotation (Figure 3A), and their muscle strength during the grip strength test was also clearly higher than that of the PD group (Figure 3B). Gait analysis (CatWalk) showed a clear increase in the average walking speed of the Y group (Figure 3C), explaining that probiotic Y7 can effectively improve motor impairments caused by PD.

[0072] In terms of non-motor functions, the fecal volume and water content of rats in group Y were significantly improved (Figures 4A, 4B), reflecting improved intestinal function. In the Morris water maze test, the latency period for searching for hidden plateaus was shortened in group Y, indicating improved spatial learning and memory abilities (Figure 4C). Furthermore, the levels of genes associated with close binding (zo1, occludin, claudin) in the rat colon were significantly elevated in group Y (Figures 4D-4F), suggesting that intestinal barrier function can be strengthened.

[0073] Serum inflammation and oxidative pressure indicators also improved. Compared to the PD group, the Y group showed a significant decrease in IL-6, IL-1β, and TNF-α levels (Figure 5A), a decrease in reactive oxygen species (ROS) concentrations, and a significant increase in the activity of antioxidant enzymes, such as superoxide dismutase (SOD) and glutathione peroxidase (GPx) (Figure 5B), indicating a reduction in systemic inflammation and oxidative pressure.

[0074] Further analysis of the enteric bacterial flora structure revealed that the Y7 group showed changes in relative abundance distribution at the genus level and exhibited a prominent group in the bacterial flora structure in the diversity index (Shannon, Simpson) and principal coordinate analysis (PCoA) diagrams (Figure 6). LEfSe and metaSeq statistical analysis indicated that Y7 significantly altered the composition of dominant and recessive bacteria (Figure 7), suggesting its potential for regulating the enteric bacterial flora and maintaining ecological equilibrium.

[0075] In summary, probiotic Y7 is a gut-brain axis functional composition with potential for clinical application, possessing multiple mechanisms of action including neuroprotection, anti-inflammatory effects, and regulation of the intestinal barrier and bacterial flora, and can simultaneously improve both motor and non-motor symptoms in PD rats.

[0076] Example 2-1: Enhancement of L-DOPA therapeutic stability and central nervous system protective effects by probiotic cocktail formulation.

[0077] This embodiment includes a combination of a specific probiotic (Lactobacillus reuteri Y7) and functional amino acids. The purpose is to evaluate the adjunctive effects of this probiotic cocktail medical food formulation ("TMU-01 formulation") on the treatment of Parkinson's disease (PD), particularly whether it can improve the stability and central availability of L-DOPA and whether it has neuroprotective effects. The experiment used a typical PD animal model in which dopamine neuronal damage was induced by unilateral injection of 6-hydroxydopamine (6-OHDA) into the medial forebrain bundle (MFB) of rats. Thirty male Sprague-Dawley rats were selected and randomly divided into six groups: a normal control group (NC group), a PD model group (PD group), an L-DOPA treatment group (PL group), an L-DOPA + benserazide group (PLI group), an L-DOPA + probiotic Y7 group (PLY group), and an L-DOPA + Y7 + TMU-01 group (PLM group). Benserazide is an aromatic L-amino acid decarboxylase inhibitor that can help improve the efficiency of L-DOPA's entry into the central nervous system by preventing its premature conversion to dopamine in the periphery. All prescriptions were administered orally via gastric injection once daily for nine consecutive weeks. Modeling was performed in week 0, behavioral testing was conducted from week 8 to week 9, and sacrificial sampling was performed in week 10 (Figure 8).

[0078] TH immunohistochemical staining showed a significant decrease in the number of TH-positive neurons in the substantia nigra pars compacta (SNpc) of the PD group, and only a small number remained in the PL group. Conversely, all three groups—PLI, PLY, and PLM—recovered a significant number of neurons, indicating that the PLM group was the most effective (Figures 9A, 9B).

[0079] In terms of athletic performance, the Rotarod test showed that the running distance and fall time were significantly longer in the PLY and PLM groups than in the PD and PL groups (Figures 10A, 10B). The grip strength test results showed that the PLM group exhibited the most significant muscle strength recovery (Figure 10C), indicating that the muscle strength of the PLM group was clearly superior to that of the other treatment groups.

[0080] Serum analysis showed that L-DOPA concentrations were significantly higher in all three groups (PLI, PLY, and PLM) than in the PL group (Figure 11C), but DA concentrations were relatively lower (Figure 11F), indicating improved L-DOPA metabolic stability. The L-DOPA / DA ratio improved in all three groups (PLI, PLY, and PLM), with the most significant improvement in PLM (Figure 11G), reflecting its highest central nervous system utilization rate.

[0081] Inflammation-related analyses revealed that inflammatory pathway genes in the PD group's colon, such as TLR4 (Figure 12A), NF-κB (Figure 12B), TNF-α (Figure 12C), and IL-6 (Figure 12D), were clearly elevated. However, these all showed a decreasing trend after treatment with PLI, PLY, and PLM, with the inhibition in the PLM group being the most pronounced.

[0082] In terms of the intestinal barrier, the expression of ZO-1, Occludin, and Claudin-1 genes was relatively high in both the PLM and PLY groups (Figures 13A-13C), with the PLM group showing the most significant improvement, indicating a clear improvement in intestinal integrity.

[0083] Summary: The results of this embodiment demonstrate that a probiotic cocktail medical food formulation can increase the bioavailability of L-DOPA, protect neurons, improve motor expression, inhibit inflammation, and strengthen the intestinal barrier. Here, the PLM group showed the most comprehensive effects and is demonstrated to be usable as an effective adjunct to L-DOPA treatment.

[0084] Example 2-2: Delay in long-term therapeutic side effects and improvement of bacterial flora with probiotic formulations.

[0085] This study aims to evaluate whether a probiotic cocktail food formulation can delay the onset of side effects associated with long-term L-DOPA treatment in Parkinson's disease, and to examine its effects on behavioral expression and central nervous system biochemical indicators. The PD rat model shown in Figure 14 was prepared using a typical method, and dopaminergic neuronal damage was induced by unilateral injection of the neurotoxin 6-hydroxydopamine (6-OHDA) into the medial forebrain bundle (MFB) of rats. For the experiment, 30 male Sprague-Dawley rats were selected and randomly divided into four groups: a normal control group (NC), a PD model + L-DOPA + Benserazide group (PDD), a PDD + probiotic Y7 group (PDDY), and a PDD + TMU-01 group (PDDM). Each group was perfused daily for 9 weeks (Figure 14).

[0086] At week 8, an Abnormal Involuntary Movements (AIMs) test was performed, and behavioral score analysis was conducted based on the Abnormal Involuntary Movements score sheet (Figure 15). The experimental results showed that the AIMs scores of rats in the PDDM group were significantly lower than those of the PDD group that received L-DOPA + Benserazide treatment alone, demonstrating that this treatment can effectively mitigate L-DOPA-induced side effects (Figure 16).

[0087] In the Morris water maze test, the PDDM group performed best in finding plateau latency and total swimming distance, and was shown to be superior to the other groups in terms of cognitive function representation, and to have the potential to significantly improve spatial learning and memory (Figures 17A-17C).

[0088] In terms of L-DOPA metabolic stability, the PDDM group had the highest serum L-DOPA concentration and L-DOPA / DA ratio (Figures 18A-18C), indicating that it can effectively stabilize L-DOPA concentrations in the body and enhance its potential for central nervous system entry and utilization.

[0089] Microbial analysis showed that the PDDM group was superior in terms of genus and door-level probiotic recovery (Figures 19A-19C), diversity index (Figure 20A), and principal coordinate analysis grouping (Figure 20B). LEfSe and LDA analysis results supported the significant changes in its microbiome characteristics (Figures 21A-B). Overall, the superiority of the PDDM group in L-DOPA metabolic stability and regulation of the gut microbiota indicates that this TMU-01 formulation possesses multiple physiological regulatory functions and therapeutic potential.

[0090] Summary: The probiotic cocktail medical food formulation PDDM exhibits significant therapeutic benefits in a rat model of L-DOPA-induced Parkinson's disease with movement disorders. PDDM effectively delays abnormal involuntary movements (AIMs) caused by long-term L-DOPA use, improves spatial memory and learning ability, stabilizes L-DOPA metabolism, and increases its bioavailability. Furthermore, PDDM promotes the balance of the gut microbiota and the restoration of probiotics, further supporting its potential as an adjunctive therapy for Parkinson's disease and highlighting the importance of gut health in PD treatment.

[0091] As described above, the probiotic cocktail medical food formulation according to the present invention has been demonstrated, based on the results of a series of animal experiments, to have a clear and comprehensive adjunctive effect in improving motor and non-motor symptoms associated with Parkinson's disease, and in enhancing the stability and safety of L-DOPA treatment. Probiotic Y7 alone can improve behavioral coordination, muscle strength, and gait expression in rats in a PD animal model, and can have a clear effect in digestive function, intestinal barrier integrity, and spatial memory ability. At the same time, Y7 has been shown to effectively inhibit systemic inflammatory responses and oxidative pressure, promote the expression of adherent proteins in the intestinal tract and the reconstruction of the probiotic bacterial community, and have a holistic regulatory function on the gut-brain axis. Furthermore, it was inferred that Y7, when used in combination with L-DOPA and after the addition of specific amino acids, can not only protect the survival of dopaminergic neurons in the central nervous system, but also significantly improve the concentration and stability of L-DOPA in serum and maintain a relatively high L-DOPA-to-dopamine concentration ratio, thus promoting the cross-section of the drug to the blood-brain injury wall and improving drug utilization. This probiotic cocktail medical food formulation can alleviate abnormal involuntary movements (LID) caused by long-term L-DOPA use and improve cognitive decline issues. Both its delayed effect on movement disorders and its maze learning expression are superior to those of the monotherapy group.

[0092] The innovative aspect of this invention lies in its focus on intestinal regulation, incorporating amino acids that influence neurotransmission and energy metabolism through probiotics with neuroprotective and anti-inflammatory effects, thereby constructing an adjunctive therapy with multi-target, multi-stage intervention capabilities. Compared to conventional drug designs that target only drug metabolism or receptor action, this invention focuses on improving the overall gut-brain axis environment, overcoming key bottlenecks such as limitations in L-DOPA therapeutic efficacy, frequent side effects, and insufficient patient tolerance. Therefore, this composition provides a highly safe, long-term-use nutritional intervention strategy that complements and synergizes with current L-DOPA therapies. It can be applied as an adjunctive therapy for clinically ill Parkinson's disease patients, or used as a preventive health measure to slow disease progression, reduce drug dosages, and improve patients' quality of life, possessing high practical value and clinical application prospects.

[0093] 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 with the Japan National Institute of Advanced Industrial Science and Technology, International Patent and Biotechnology Center (IPOD, NITE) (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. An application for producing a composition that alleviates or improves the symptoms of Parkinson's disease, wherein the composition comprises at least one probiotic having intestinal regulatory or neuroprotective function.

2. The use according to claim 1, wherein the composition further comprises a combination of functional amino acids including four or more essential amino acids.

3. The use according to claim 1, further comprising extending the stability of administration of a drug effective for Parkinson's disease and reducing side effects of administration.

4. The use of the drug effective for Parkinson's disease described above, which extends the stability of administration and reduces side effects, is to increase the central availability of levodopa (L-DOPA), delay abnormal involuntary movements (L-DOPA-induced dyskinesia, LID), improve non-motor symptoms, reduce inflammation and oxidative stress responses, strengthen the integrity of the intestinal barrier, or improve the balance of the intestinal bacterial community, non-motor symptoms, and cognitive symptoms, as described in claim 1.

5. The use according to claim 3, wherein the drug effective for Parkinson's disease is selected from the group consisting of levodopa (L-DOPA), dopamine agonists, MAO-B inhibitors, DOPA decarboxylase inhibitors, and COMT inhibitors.

6. The use according to claim 1, wherein the probiotic is selected from the group consisting of Lactobacillus, Lactococcus, Streptococcus, Bifidobacterium, Pediococcus, and Enterococcus genera.

7. 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 use according to claim 1, selected from the group consisting of L. apis, L. ghanenis, L. dextrinicus, L. shenzhenensis, and L. harbinensis.

8. The use according to claim 1, wherein the probiotic is live bacteria, dead bacteria, pasteurized bacteria, degradation products, bacterial supernatant, cell wall extract, extracellular polysaccharide or its immunostimulant component.

9. The use described in claim 3, wherein the drug effective in treating Parkinson's disease is levodopa (L-DOPA).

10. The use according to claim 1, wherein the essential amino acid is selected from leucine, isoleucine, valine, lysine, methionine, phenylalanine, tryptophan, threonine, and histidine.

11. The use according to claim 4, wherein the non-motor symptoms include cognitive impairment, gastrointestinal dysfunction, or sleep disorders.

12. The composition, through overall gut-brain axis regulation and neuroprotective effects, improves Parkinson's disease behavioral function and delays L-DOPA-induced abnormal involuntary movements, as described in claim 1.

13. The composition, in its use according to claim 1, achieves improvement of motor and non-motor symptoms of Parkinson's disease and reduction of L-DOPA side effects by regulating the intestinal bacterial flora, reducing systemic inflammation, and improving the central availability of L-DOPA.

14. The use according to claim 1, wherein the Parkinson's disease is in the prodromal, early or late stage of Parkinson's disease, or the person has not received treatment.

15. A composition for improving the symptoms of Parkinson's disease, extending the stability of administration of drugs effective against Parkinson's disease, and reducing side effects of administration, comprising: (a) at least one probiotic bacterium having intestinal regulatory or neuroprotective function; and (2) a combination of functional amino acids including four or more essential amino acids.

16. The composition according to claim 15, wherein the agent effective in treating Parkinson's disease is selected from the group consisting of levodopa (L-DOPA), dopamine agonists, MAO-B inhibitors, DOPA decarboxylase inhibitors, and COMT inhibitors.

17. The composition according to claim 15, wherein the drug effective in treating the aforementioned Parkinson's disease is levodopa (L-DOPA).

18. The composition according to claim 15, wherein the probiotic is selected from the genera Lactobacillus, Bifidobacterium, or Streptococcus.

19. The composition according to claim 15, wherein the probiotics are live bacteria, dead bacteria, pasteurized bacteria, degradation products, bacterial supernatant, cell wall extracts, extracellular polysaccharides or their immunostimulant components.

20. The composition according to claim 15, wherein the essential amino acid is selected from leucine, isoleucine, valine, lysine, methionine, phenylalanine, tryptophan, threonine, and histidine.

21. A pharmaceutical composition comprising (a) an agent effective in treating Parkinson's disease, (b) at least one probiotic bacterium having tumor-regulating or neuroprotective functions, and (c) a combination of functional amino acids containing four or more essential amino acids.

22. The pharmaceutical composition according to claim 21, wherein the drug effective in treating Parkinson's disease is levodopa (L-DOPA).

23. The pharmaceutical composition according to claim 22, wherein the probiotic is L. reuteri.

24. The pharmaceutical composition according to claim 22, wherein the essential amino acid is selected from leucine, isoleucine, valine, lysine, methionine, phenylalanine, tryptophan, threonine, and histidine.

25. The pharmaceutical composition according to claim 21, wherein the pharmaceutical composition can extend the stability of administration of a drug effective against Parkinson's disease and reduce side effects of administration.

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