Treatment of Alzheimer's disease with allogenic mesenchymal stem cells

Abstract

This provides methods to alleviate the symptoms of Alzheimer's disease (AD). [Solution] Composition and method for treating Alzheimer's disease with allogenic mesenchymal stem cells. The treatment method involves administering a composition of allogenic mesenchymal stem cells to a subject in need, and the effectiveness of the treatment method can be determined by measuring specific biomarkers and improving cognitive function.

Description

[Technical Field]

[0001] [Cross-reference of related applications] This application relates to and claims the benefits of U.S. Provisional Patent Application No. 63 / 075,686, “Therapy of Alzheimer’s Disease by Allogenic Mesenchymal Stem Cells,” filed on 8 September 2020, No. 63 / 134,535, “Therapy of Alzheimer’s Disease by Allogenic Mesenchymal Stem Cells,” filed on 6 January 2021, and No. 63 / 173,960, “Therapy of Alzheimer’s Disease by Allogenic Mesenchymal Stem Cells,” filed on 12 April 2021, the entire contents of which are incorporated herein by reference in their entirety.

[0002] [Field] This application relates to methods and compositions for the treatment of Alzheimer's disease in subjects requiring treatment for Alzheimer's disease. Some embodiments relate to compositions comprising a therapeutically effective amount of allogeneic mesenchymal stem cells (MSCs) used to alleviate symptoms of Alzheimer's disease, such as increased systemic inflammation. Other embodiments relate to a treatment method in which a subject suffering from symptoms of Alzheimer's disease is administered a therapeutically effective amount of a composition comprising MSCs. The effectiveness of these treatments is evaluated by measuring the concentration of specific biomarkers in the subject after administration of the MSC-containing composition, examining changes in brain activity or morphology, and determining whether cognitive function has improved after treatment.

[0003] [background] Alzheimer's disease (AD) is characterized by a complex pathogenesis involving diverse mechanisms in addition to β-amyloid deposition and neurofibrillary entanglement [1]. There is a growing recognition that pro-inflammatory states contribute to the subsequent development of dementia [2-4]. In this regard, pro-inflammatory cytokines are abundant near amyloid deposits and neurofibrillary entanglement [5], and there is a link between systemic inflammation and β-amyloid accumulation [4]. AD is further characterized by neurovascular disorders that contribute to adverse outcomes [6]. The resulting blood-brain barrier (BBB) ​​disruption [7-10] impairs endothelial exchange, which may lead to inefficient clearance and accumulation of AβP in the brain [11, 12].

[0004] Due to the complex nature of AD progression, predicting the onset and progression of AD using biomarkers remains difficult. While β-amyloid deposition and neurofibrillary tangle concentrations can be used to diagnose or predict the onset of AD, studies have shown that some individuals with significant amyloid deposition and neurofibrillary tangles at autopsy, and deemed worthy of an AD diagnosis, have no history of dementia. Currently approved treatments for Alzheimer's disease (rivastigmine, donepezil, memantine, galantamine, tacrine) offer only minimal benefits as they are primarily symptomatic treatments; furthermore, no approved therapies can effectively stop, reverse, or prevent AD. The consistent failure of initially promising lead compounds has resulted in the lack of approval of new AD drugs for over a decade. Recently, the failures of the anti-amyloid monoclonal antibodies solanezumab (Ely Lily) and aducanumab (Biogen / Eisai) in mild to moderate Alzheimer's disease (AD) and mild cognitive impairment (MCI) have been identified. A common theme in these failures is that they target a single pathological feature of AD.

[0005] Addressing these neuropathological features of AD simultaneously could offer therapeutic benefits and lead to new therapeutic strategies. Medicinal signaling cells (MSCs, also known as mesenchymal stem cells) are pluripotent cells (in vitro) with a multifaceted mechanism of action (MOA), including anti-inflammatory properties, the ability to improve vascular function, and the promotion of endogenous tissue repair and regeneration [13, 14]. MSCs can migrate to inflammatory and injury sites, making them potentially effective targets for neuroinflammatory sites in AD. MSCs can also modulate the host stem cell niche through paracrine activity and heterocellular junction, promoting endogenous repair and regeneration

[15] . Finally, MSCs are immune-evading / immunely privileged, allogenically usable, and possess an acceptable safety profile in clinical trials. These immune-privileged / immunely evasive properties mean that MSCs have undetectable levels of major histocompatibility complex class II (MHC-II) molecules and low levels of MHC-I, making them a potentially readily available "off-the-shelf" therapeutic option for a broad patient population.

[0006] Several preclinical data support the efficacy of MSCs in AD. In animal models, MSCs cross the blood-brain barrier, promote neurogenesis, inhibit β-amyloid deposition and promote clearance, reduce apoptosis, promote hippocampal neurogenesis, improve dendritic morphology, and improve behavioral and spatial memory performance [18-20]. These beneficial effects were associated with reduced inflammation, increased Aβ degradation factors and Aβ clearance, decreased hyperphosphorylated tau, and elevated surrogate-activated microglia markers. These benefits appear to be at least partially due to Aβ-inducible MSC release by chemoattractants that recruit surrogate microglia to the brain to reduce Aβ deposition

[21] . MSCs have been reported to be effective in young AD model mice before Aβ accumulation, resulting in a significant reduction in Aβ deposition in the brain and a significant increase in presynaptic protein expression

[22] . Interestingly, these effects persisted for at least two months, suggesting that MSCs may be potentially effective as an interventional treatment for prodromal AD.

[0007] Therefore, this application aims not only to provide a method for treating AD that includes the use of a composition containing MSCs, but also to provide a method for accurately measuring the potential safety of MSCs and evaluating their efficacy in alleviating AD symptoms in subjects who require them.

[0008] [overview] The object of this application is to provide a method for treating or alleviating Alzheimer's disease (AD), comprising the step of administering a therapeutic dose of allogenic MSCs to a subject in need to alleviate the symptoms of AD and / or treat the progression of AD. Another object of this application is to provide novel biomarkers for diagnosing and evaluating the progression of AD and the effectiveness of treatment methods. These biomarkers may be changes in size in areas of the patient's brain, such as the amygdala, cortical nuclei, hippocampus, hippocampal microregions, and / or corticoamygdaloid transition.

[0009] In some embodiments, novel biomarkers for diagnosing and evaluating the progression of AD may be changes in cytokine concentrations, where the cytokines may be IL-4, IL-6, IL-8, IL-10, IL-12p70, IL-17, sIL-2Rα, or combinations thereof. In preferred embodiments, cytokine concentrations increase in the serum, plasma, cerebrospinal fluid, or blood of subjects suffering from and needing AD symptoms after administration of allogenic MSCs to the subject. The increase in cytokine concentration may be in the range of 0% to 10%, 0.5% to 10%, 1.0% to 10%, 3% to 10%, 5% to 10%, 7% to 10%, greater than 0% to less than 10%, 10% to 50%, 20% to 50%, 30% to 50%, or greater than 50%. In a preferred embodiment, the cytokine concentration reaches and maintains a different concentration level from the concentration level before MSC administration to the target requiring it, and then increases to a stable concentration level that does not decrease beyond 0%-10%, 0%-5%, or 0%-1%.

[0010] In other embodiments, novel biomarkers for diagnosing and evaluating the progression of AD may be changes in the concentration of neuron-related molecules or peptides, where the neuron signaling molecules or peptides may be tau, phosphotau, Aβ-38, Aβ-40, Aβ-42, NFL, or combinations thereof. In preferred embodiments, the concentration of Aβ-38, Aβ-40, or Aβ-42 increases in the serum, plasma, or blood of a subject suffering from AD symptoms and requiring it, after administration of allogenic MSCs to the subject. The increase in Aβ-38, Aβ-40, or Aβ-42 concentration may be in the range of 0% to 10%, 0.5% to 10%, 1.0% to 10%, 3% to 10%, 5% to 10%, 7% to 10%, greater than 0% to less than 10%, 10% to 50%, 20% to 50%, 30% to 50%, or greater than 50%. In a preferred embodiment, the concentration of Aβ-38, Aβ-40, or Aβ-42 reaches and maintains a different concentration level from the concentration level before the administration of MSCs to the subject requiring it, and then increases to a stable concentration level that does not decrease beyond 0% to 10%, 0% to 5%, or 0% to 1%.

[0011] In other embodiments, the concentration of tau, phosphotau, or NFL decreases in the serum, plasma, cerebrospinal fluid, or blood of a subject suffering from AD symptoms and requiring it after administration of allogenic MSCs to the subject. The decrease in tau, phosphotau, or NFL concentration may be in the range of 0% to 10%, 0.5% to 10%, 1.0% to 10%, 3% to 10%, 5% to 10%, 7% to 10%, greater than 0% to 10%, 10% to 50%, 20% to 50%, 30% to 50%, or greater than 50%. In preferred embodiments, the concentration of tau, phosphotau, or NFL reaches and maintains a concentration level different from the concentration level before administration of MSCs to the subject requiring it, and then decreases to a stable concentration level that does not rise above 0% to 10%, 0% to 5%, or 0% to 1%.

[0012] In other embodiments, novel biomarkers for diagnosing and evaluating the progression of AD may be changes in the concentration of inflammatory signaling molecules, which may be pro-BNP, TNF-α, or a combination thereof. In preferred embodiments, the concentration of TNF-α or pro-BNP decreases in the serum, plasma, cerebrospinal fluid, or blood of a subject suffering from AD symptoms and in need of it, after administration of allogenic MSCs to the subject. The decrease in TNF-α or pro-BNP concentration may be in the range of 0% to 10%, 0.5% to 10%, 1.0% to 10%, 3% to 10%, 5% to 10%, 7% to 10%, greater than 0% to 10%, 10% to 50%, 20% to 50%, 30% to 50%, or greater than 50%. In a preferred embodiment, the concentration of TNF-α or pro-BNP reaches and maintains a different concentration level from the concentration level before MSC administration to the subject in need, and then decreases to a stable concentration level that does not rise above 0%-10%, 0%-5%, or 0%-1%.

[0013] In some embodiments, novel biomarkers for diagnosing and evaluating the progression of AD may be changes in VEGF concentration and other vascular-related biomarkers. In preferred embodiments, VEGF concentration increases in the serum, plasma, cerebrospinal fluid, or blood of a subject suffering from AD symptoms and in need of it, after administration of allogenic MSCs to the subject. The increase in VEGF concentration may be in the range of 0% to 10%, 0.5% to 10%, 1.0% to 10%, 3% to 10%, 5% to 10%, 7% to 10%, greater than 0% to 10%, 10% to 50%, 20% to 50%, 30% to 50%, or greater than 50%. In preferred embodiments, VEGF concentration increases to a stable concentration level that does not decrease by more than 0% to 10%, 0% to 5%, or 0% to 1%, once it reaches and maintains a different concentration level than the concentration level before administration of MSCs to the subject in need of it.

[0014] Allogenic MSCs may also be LOMECEL-B® cells, which are Longeveron formulations of allogenic human mesenchymal stem cells. Further uses and formulations of useful stem cells, including LOMECEL-B® branded mesenchymal cells, can be found in the following U.S. Patent Application Publications, all of which are incorporated herein by reference: U.S. Patent Application No. 20190038742A1; U.S. Patent Application No. 20190290698A1; and U.S. Patent Application No. 20200129558A1. [Brief explanation of the drawing]

[0015] [Figure 1] This figure illustrates a phase 1, double-blind, randomized, placebo-controlled clinical trial conducted to determine the efficacy of lomesel-B in the treatment of patients diagnosed with mild Alzheimer's disease (AD). [Figure 2] This diagram shows the experimental groups used in the Phase 1 clinical trial and how the subjects were divided into each experimental group. [Figure 3A] Figure 3A shows the MMSE scores of three experimental groups (20 × 10⁶ Lomesel-B, 100 × 10⁶ Lomesel-B, and placebo) over a 6-month period. [Figure 3B] Figure 3B shows the changes in ADAS-cog scores over a 6-month period in three experimental groups (20 × 10⁶ Lomesel-B cells, 100 × 10⁶ Lomesel-B cells, and placebo). [Figure 3C] Figure 3C shows the changes in TMT-A scores over a 6-month period in three experimental groups (20 × 10⁶ Lomesel-B cells, 100 × 10⁶ Lomesel-B cells, and placebo). [Figure 3D] Figure 3D shows the changes in TMT-B scores over a 6-month period in three experimental groups (20 × 10⁶ Lomesel-B cells, 100 × 10⁶ Lomesel-B cells, and placebo). [Figure 3E] Figure 3E shows the changes in GDS points over a 6-month period in three experimental groups (20 × 10⁶ Lomesel-B cells, 100 × 10⁶ Lomesel-B cells, and placebo). [Figure 4A] Fig. 4A is a diagram showing changes in the subject version of QOL-AD scores over 6 months for three experimental groups (20×10^6 cells of Lomecel-B, 100×10^6 cells of Lomecel-B and placebo). [Figure 4B] Fig. 4B is a diagram showing changes in the caregiver version of QOL-AD scores over 6 months for three experimental groups (20×10^6 cells of Lomecel-B, 100×10^6 cells of Lomecel-B and placebo). [Figure 4C] Fig. 4C is a diagram showing changes in ADCS-ADL scores over 6 months for three experimental groups (20×10^6 cells of Lomecel-B, 100×10^6 cells of Lomecel-B and placebo). [Figure 4D] Fig. 4D is a diagram showing changes in ADRQL scores over 6 months for three experimental groups (20×10^6 cells of Lomecel-B, 100×10^6 cells of Lomecel-B and placebo). [Figure 5A] Fig. 5A is a diagram showing changes in relative VEGF concentration in serum over 6 months for three experimental groups (20×10^6 cells of Lomecel-B, 100×10^6 cells of Lomecel-B and placebo). [Figure 5B] Fig. 5B is a diagram showing changes in relative IL-4 concentration in serum over 6 months for three experimental groups (20×10^6 cells of Lomecel-B, 100×10^6 cells of Lomecel-B and placebo). [Figure 5C] Fig. 5C is a diagram showing changes in relative IL-6 concentration in serum over 6 months for three experimental groups (20×10^6 cells of Lomecel-B, 100×10^6 cells of Lomecel-B and placebo). [Figure 5D] Fig. 5D is a diagram showing changes in relative sIL-2Rα concentration in serum over 6 months for three experimental groups (20×10^6 cells of Lomecel-B, 100×10^6 cells of Lomecel-B and placebo). [Figure 5E] Fig. 5E is a diagram showing changes in relative IL-10 concentration in serum over 6 months for three experimental groups (20×10^6 cells of Lomecel-B, 100×10^6 cells of Lomecel-B and placebo). [Figure 5F] Figure 5F shows the relative changes in serum IL-12 concentration over a 6-month period in three experimental groups (20 × 10⁶ Lomecell-B, 100 × 10⁶ Lomecell-B, and placebo). [Figure 6A] Figure 6A shows the changes in brain volume in the left hippocampal region over a 6-month period in three experimental groups (20 × 10⁶ Lomesel-B cells, 100 × 10⁶ Lomesel-B cells, and placebo). [Figure 6B] Figure 6B shows the changes in brain volume in the right hippocampal region over a 6-month period in three experimental groups (20 × 10⁶ Lomesel-B cells, 100 × 10⁶ Lomesel-B cells, and placebo).

[0016] [Detailed explanation] There is only one FDA-approved disease-modifying intervention for Alzheimer's disease (AD) (aducanumab), but this is controversial and may slow the progression of dementia in a subpopulation of AD patients. Other FDA-approved AD treatments are symptomatic only and do not alter disease progression. The onset and development of AD are still not fully understood, and this uncertainty is the source of much controversy in this field. In fact, while the majority of researchers in the field of Alzheimer's disease onset and treatment acknowledge that AβP accumulation is involved in disease progression, it is still unknown whether AβP accumulation is the cause of Alzheimer's disease, or whether it is simply a result of dysregulation of other cellular pathways as a consequence of aging.

[0017] Patients with Alzheimer's disease (AD) are known to trigger irregular immune responses. Indeed, the presence of abundant pro-inflammatory cytokines near amyloid deposits and neurofibrillary entanglements has suggested a link between systemic inflammation and β-amyloid accumulation. In light of this evidence, those skilled in the art have often been skeptical about the role of the immune system in the development of AD, given the lack of a direct correlation between the inhibition of pro-inflammatory cytokines and the reduction of AβP accumulation in humans.

[0018] Therefore, we were surprised to discover that the use of compositions containing allogenic MSCs can combat the symptoms of AD. When subjects suffering from AD symptoms were treated with compositions containing allogenic stem cells, we found that the morphology of the subjects' brains improved and the expression of biomarkers related to anti-inflammatory and vascular repair was promoted. We also found that allogenic MSCs can promote the improvement of neuroinflammation and vascular function in subjects suffering from AD symptoms. These findings are surprising because those skilled in the art had generally refrained from using MSCs in the treatment of AD due to the ambiguity surrounding the pathogenesis of AD, as well as the fact that MSCs cannot directly target β-amyloid and have a short residence time in the human body, leading to the expectation that using MSCs would not be effective. Furthermore, those skilled in the art expected that the efficacy of AD treatment would be low because they believed that due to their large size, they could not cross the blood-brain barrier and reach the sites of inflammation and injury.

[0019] Another advantage of using MSCs in the treatment of AD is that they do not involve targeting a single pathway or a biomarker such as AβP accumulation. Instead, by using MSCs in the treatment of AD, multiple pathways can be targeted at once, which can stop or significantly slow the progression of AD.

[0020] Following the remarkable findings described above, one aspect of this application relates to a method for treating or alleviating the symptoms of Alzheimer's disease, comprising the step of administering a composition containing allogenic MSCs to a subject suffering from symptoms of Alzheimer's disease.

[0021] In some embodiments, a method for treating AD or alleviating the symptoms of AD further includes the step of measuring the concentration of a biomarker in a subject suffering from symptoms of AD before and / or after administration of a composition comprising allogenic MSCs.

[0022] In other embodiments, a method for treating AD or alleviating the symptoms of AD further includes the step of measuring the cognitive function of a subject before and / or after administration of a composition comprising allogenic MSCs.

[0023] In some embodiments, the MSC used in the treatment method is Lomecel-B (trademark) MSC.

[0024] In other embodiments, the biomarker is a cytokine such as IL-4, IL-6, IL-8, IL-10, IL-12p70, IL-17, sIL-2Rα, or a combination thereof.

[0025] In a preferred embodiment, cytokine concentrations increase in the serum, plasma, cerebrospinal fluid, or blood of a subject suffering from AD symptoms and in need of it, after administration of allogenic MSCs to the subject. The increase in cytokine concentration may be in the range of 0% to 10%, 0.5% to 10%, 1.0% to 10%, 3% to 10%, 5% to 10%, 7% to 10%, greater than 0% to 10%, 10% to 50%, 20% to 50%, 30% to 50%, or greater than 50%. In a preferred embodiment, cytokine concentrations reach and are maintained at a concentration level different from the concentration level before administration of MSCs to the subject in need, and then increase to a stable concentration level that does not decrease by more than 0% to 10%, 0% to 5%, or 0% to 1%.

[0026] In other embodiments, the biomarker is a neuron-related molecule or peptide such as tau, phosphotau, Aβ-38, Aβ-40, Aβ-42, NFL, or a combination thereof.

[0027] In a preferred embodiment, the concentration of Aβ-38, Aβ-40, or Aβ-42 increases in the serum, plasma, or blood of a subject suffering from AD symptoms and in need of it, after administration of allogenic MSCs to the subject. The increase in the concentration of Aβ-38, Aβ-40, or Aβ-42 may be in the range of 0% to 10%, 0.5% to 10%, 1.0% to 10%, 3% to 10%, 5% to 10%, 7% to 10%, greater than 0% to 10%, 10% to 50%, 20% to 50%, 30% to 50%, or greater than 50%. In another embodiment, the concentration of Aβ-38, Aβ-40, or Aβ-42 increases to a stable concentration level that does not decrease by more than 0% to 10%, 0% to 5%, or 0% to 1%, once it reaches and maintains a different concentration level than the concentration level before administration of MSCs to the subject in need of it.

[0028] In other embodiments, the concentration of tau, phosphotau, or NFL decreases in the serum, plasma, cerebrospinal fluid, or blood of a subject suffering from AD symptoms and requiring it, after administration of allogenic MSCs to the subject. The decrease in tau, phosphotau, or NFL concentration may be in the range of 0% to 10%, 0.5% to 10%, 1.0% to 10%, 3% to 10%, 5% to 10%, 7% to 10%, greater than 0% to 10%, 10% to 50%, 20% to 50%, 30% to 50%, or greater than 50%. In other embodiments, the concentration of tau, phosphotau, or NFL reaches and maintains a concentration level different from the concentration level before administration of MSCs to the subject requiring it, and then decreases to a stable concentration level that does not increase by more than 0% to 10%, 0% to 5%, or 0% to 1%.

[0029] In other embodiments, the biomarker is an inflammation signaling molecule such as pro-BNP, TNF-α, or a combination thereof.

[0030] In a preferred embodiment, the concentration of TNF-α or pro-BNP decreases in the serum, plasma, cerebrospinal fluid, or blood of a subject suffering from AD symptoms and in need of it, after administration of allogenic MSCs to the subject. The decrease in TNF-α or pro-BNP concentration may be in the range of 0% to 10%, 0.5% to 10%, 1.0% to 10%, 3% to 10%, 5% to 10%, 7% to 10%, greater than 0% to 10%, 10% to 50%, 20% to 50%, 30% to 50%, or greater than 50%. In other embodiments, the concentration of TNF-α or pro-BNP reaches and maintains a different concentration level from the concentration level before administration of MSCs to the subject in need of it, and then decreases to a stable concentration level that does not increase by more than 0% to 10%, 0% to 5%, or 0% to 1%.

[0031] In some embodiments, the biomarker is VEGF or other vascular-related biomarkers.

[0032] In a preferred embodiment, the concentration of VEGF increases in the serum, plasma, cerebrospinal fluid, or blood of a subject suffering from AD symptoms and in need of it, after administration of allogenic MSCs to the subject. The increase in VEGF concentration may be in the range of 0% to 10%, 0.5% to 10%, 1.0% to 10%, 3% to 10%, 5% to 10%, 7% to 10%, greater than 0% to 10%, 10% to 50%, 20% to 50%, 30% to 50%, or greater than 50%. In a preferred embodiment, the concentration of VEGF reaches and maintains a different concentration level from the concentration level before administration of MSCs to the subject in need of it, and increases to a stable concentration level that does not decrease by more than 0% to 10%, 0% to 5%, or 0% to 1%.

[0033] In other embodiments, a method for treating AD or alleviating the symptoms of AD further includes the step of determining a change in the size of a target brain region after administration of a composition comprising allogenic MSCs. The target brain region that changes in size may be the amygdala, cortical nucleus, hippocampus, or other structure.

[0034] In another embodiment, a method for treating AD or alleviating the symptoms of AD further includes the step of examining the cerebrospinal fluid of the subject before and after administration of a composition comprising allogenic MSCs.

[0035] In another embodiment, a method for treating AD or alleviating the symptoms of AD further includes the step of examining the serum of the subject before and after administration of a composition comprising allogenic MSCs.

[0036] In another embodiment, a method for treating AD or alleviating the symptoms of AD further includes the step of examining the plasma of the subject before and after administration of a composition comprising allogenic MSCs.

[0037] In some embodiments, a method for treating AD or alleviating the symptoms of AD further includes the step of determining whether changes have occurred in the target corticoamygdal junction after administration of a composition comprising allogenic MSCs.

[0038] In other embodiments, the composition is 20 × 10 6 Individual MSC, 100 x 10 6 Individual MSCs, or 20 × 10 6 100 x 10 6 It may contain any of the following MSCs:

[0039] [Examples] Example 1: A double-blind Phase I clinical trial evaluating the efficacy of MSCs in the treatment of AD symptoms. Experimental design: The Phase 1 trial was a double-blind, randomized, and placebo-controlled trial (Figure 1), registered on ClinicalTrials.gov (NCT02600130), and monitored by a single Institutional Review Board, an independent Data and Safety Monitoring Board (DSMB), an independent clinical monitor, and the Food and Drug Administration (FDA) based on an Investigational New Drug Application (IND). All subjects and caregivers consented to participate in the trial. Subject screening consisted of a three-step process: a clinical assessment for the possibility of mild AD, MRI to rule out confounding factors, and an amyloid tracer PET scan to confirm a mild AD diagnosis. Enrolled subjects received low-dose lomesel-B [2.0 × 10⁶]. 7 Cells ("20M")), high dose of Lomesel-B [(1.0 × 10 8 Participants were randomly assigned to receive either a single infusion of cells ("100M") or a placebo. Upon completion of enrollment, 33 participants were enrolled (compared to an expected 30), including all eligible participants who had attended the screening. The infusion day was defined as day 0. Follow-up surveys were conducted at weeks 2, 4, 13, 26, 39, and 52 post-infusion.

[0040] Romesel-B and placebo: Lomecel-B is a formulation prepared by culturing and growing allogenic MSCs (Medical Stem Cells) collected from healthy young adult donors in accordance with Federal Regulation 1271 code, under current Good Manufacturing Practices (cGMP) and the FDA-approved IND's Chemical, Manufacturing, and Control (CMC) section. The placebo consisted of a vehicle (PlasmaLyte-A containing 1% human serum albumin) in which Lomecel-B MSCs were resuspended. Lomecel-B and placebo were prepared in identically labeled and identically-looking infusion bags and delivered by peripheral intravenous infusion in an outpatient setting.

[0041] Clinical assessment: Clinical assessments were performed at baseline and at weeks 2, 13, 26, 39, and 52, but the MMSE

[24] was administered at the screening visit (eligibility criteria) instead of at the baseline visit. The clinical assessments used were the 11-item Alzheimer's Disease Assessment Scale-Cognitive Subscale (ADAS-Cog), Trail Making Test Parts A and B (TMT-A and TMT-B), Neuropsychiatric Inventory (NPI), short version of the Geriatric Depression Scale (GDS), ADCS-ADL, Alzheimer's Disease-Related Quality of Life (ADRQL), a caregiver self-assessment questionnaire developed by the American Medical Association, and the patient version and caregiver version of QOL-AD.

[0042] Biomarkers: Assays for vascular endothelial growth factor (VEGF), D-dimer, N-terminal pro-B-type natriuretic peptide, transforming growth factor-β1, C-reactive protein, interleukin (IL-)5, IL-17, and soluble IL-2Rα (sIL-2Rα) were performed by a central laboratory (Cenetron Diagnostics: Austin, Texas). High-sensitivity electrochemiluminescence immunoassays for IL-1β, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12p70, IL-13, tumor necrosis factor α (TNF-α), TNF-α-stimulated gene 6, interferon gamma, amyloid beta (Aβ) peptide 1-38 (Aβ 38 ), Aβ 40 , Aβ 42 , total tau, phospho-tau T181, and neurofilament light chain (NfL) were performed by Longeveron using the MESO QuickPlex SQ 120 system (Meso Scale Diagnostics, LLC: Rockville, Maryland). CSF collection was optional in this safety study, and samples were limited, so formal statistical analysis could not be performed.

[0043] Brain MRI was performed at screening, and at weeks 13, 26, 39, and 52 to assess safety (including ARIA), and was further used to assess structural brain changes.

[0044] Screening PET includes florbetaben (18 F ) was used (Life Molecular Imaging: Boston, Massachusetts). Patients who had a positive amyloid tracer PET scan prior to screening were permitted to enroll without requiring this scan (if they met the enrollment criteria).

[0045] Statistical analysis: The sample size was selected to detect adverse events (AEs) occurring in 5% or more of the study population with a 79% probability. The analysis was performed by an independent third-party statistical group. Statistical tests were performed at a 0.05 significance level using two-tailed tests, and 95% confidence intervals were calculated as needed. No adjustments were made for multiple analyses.

[0046] The primary endpoint was the activation of a Bayesian-motivated safety termination rule for the occurrence of a serious adverse event (SAE) (defined as a serious adverse event occurring under treatment, or TE-SAE) within the first 30 days after infusion. The threshold was calculated based on an estimated TE-SAE rate of 10.0%, and the termination rule was activated when the TE-SAE rate exceeded 40%. The termination rule had a power of 91% with a 19% probability of a Type I error.

[0047] Additional safety assessments included the following, and AEs and SAEs were evaluated throughout the study. Clinical laboratory tests (hematology, blood chemistry, coagulation, and urinalysis) were performed at screening, baseline, and infusion visits, as well as at weeks 4, 13, 26, 39, and 52. Physical and neurological examinations were also performed. Electrocardiograms (ECGs) were performed at screening and infusion visits, as well as at weeks 4 and 52.

[0048] Overall follow-up compliance was 100% up to week 13 post-infusion and 85% up to week 26 [13 out of 15 patients (87%) in the low-dose lomesel-B group, 8 out of 10 patients (80%) in the high-dose lomesel-B group, and 7 out of 8 patients (88%) in the placebo group]. Subsequently, follow-up compliance declined, with 5 patients (33%) in the low-dose lomesel-B group, 6 patients (60%) in the high-dose lomesel-B group, and 2 patients (13%) in the placebo group discontinuing before the 52-week follow-up visit (overall compliance of 61% at 52 weeks). Of the 13 discontinuances, 6 (46%) occurred during the COVID-19 pandemic. Therefore, efficacy is only provided up to week 26.

[0049] Research population: Fifty subjects were screened at four clinical sites, and 33 (66%) were enrolled and randomized between November 3, 2016 and September 19, 2019 (Figure 2). The two main reasons for screening failure were negative amyloid tracer PET scan (52% of screening failures) or concurrent MRI findings (29% of screening failures). Enrolled subjects received a single intravenous infusion of 20M lomesel-B (N=15), 100M lomesel-B (N=10), or placebo (N=8).

[0050] Baseline population statistics are shown in Table 1. The mean age was 71.2 ± 8.4 years, and 48.5% were female. At least 19 subjects (57.6%) carried at least one ApoE4 allele (2 subjects declined genetic testing).

[0051] [Table 1] TIFF2026143531000003.tif207149TIFF2026143531000004.tif118149

[0052] Primary evaluation criteria - Safety: The primary endpoint was the activation of the TE-SAE cessation rule. The cessation rule was never activated, and the primary safety endpoint was met. Only one TE-SAE occurred, in the 100M lomesel-B group, resulting in a 24-hour hospitalization due to back pain on day 27 post-infusion (Table 2), but this was determined to be unrelated to the study drug. The incidence of AEs within 30 days post-infusion (AEs occurring under treatment, i.e., TE-AEs) in the lomesel-B group was no different from that in the placebo group (16.0% of subjects in the combined lomesel-B group compared to 25.0% of subjects in the placebo group, p<0.1606).

[0053] [Table 2]

[0054] There were no adverse events (AEs) or serious adverse events (SAEs) determined to be related to the study drug. The incidence of SAEs in the study was lower in each Lomecel-B treatment group compared to the placebo group (37.5% of subjects in the placebo group compared to 16.0% of subjects in the combined Lomecel-B group). However, three of these SAEs occurred before infusion (all in the placebo group). There was one death during the study, which occurred 144 days after infusion in the 100M Lomecel-B group. The incidence of AEs was lower in the Lomecel-B groups compared to placebo (87.0% of subjects in the placebo group compared to 60.0% of subjects in the combined Lomecel-B group). Only one subject experienced a severe AE (back pain), which was in the high-dose Lomecel-B group.

[0055] The infusion was not interrupted or prematurely discontinued, and there were no associated adverse events (AEs) or adverse emergencies (SAEs). MRI evaluation revealed no reported ARIA events. Hematological, coagulation, blood chemistry, vital signs, urinalysis, and ECG data were assessed by independent pharmacovigilance monitors and DSMBs, and no trends or causes of concern were observed.

[0056] Neurocognitive and neuropsychiatric assessment: Neurocognitive and neuropsychiatric assessments were evaluated as pre-specified secondary endpoints. In the 20M Romesel-B group, the decline in MMSE scores was significantly slower compared to placebo (Figure 3A). The placebo group's score decreased by 2.99 ± 1.12 points at week 13 (p=0.0337; two-sided 95% CI -5.84 to -0.31). In contrast, the 20M Romesel-B group showed no significant change from baseline, and the difference from placebo at week 13 was significantly higher (better) by 2.69 ± 1.39 points (p=0.0182; two-sided 95% CI 0.51 to 4.97). In the 100M Romesel-B group, a trend toward a decrease in MMSE scores was shown from baseline, but not statistically significant, and there was no statistically significant difference compared to placebo.

[0057] In ADAS-cog-11, the placebo group showed a tendency towards worsening (increase) (Figure 3B). Although the lomesel-B group appeared more stable, these results were not statistically significant compared to the placebo group.

[0058] No significant change from baseline was observed in any of the TMT-A treatment groups, and there was no difference between the lomesel-B treatment group and the placebo group (Figure 3C). For TMT-B, the placebo group showed a worsening trend (longer completion time), while both lomesel-B treatment groups showed an improving trend, but the difference was not statistically significant (Figure 3D).

[0059] In the GDS study, no significant difference from baseline was observed in any of the treatment groups, nor in the Lomesel-B group compared to placebo (Figure 3E).

[0060] Assessment of quality of life and activities of daily living: In the patient version of QOL-AD, the 20M Lomecel-B group showed a significant improvement of 3.85 ± 1.943 points compared to placebo at week 26 (p=0.0444; two-sided 95% CI 0.13~9.12) (Figure 4A). There was no significant difference between the 100M Lomecel-B and placebo groups, and there was no significant change from baseline in either group. In the caregiver version of QOL-AD, all groups showed an improving trend, with the placebo group showing a significant change from baseline at week 2 (3.9 ± 4.61 points; p=0.0491; 95% CI 0.02~7.73) (Figure 4B). Compared to the change in placebo, there was no significant change from baseline in the Lomecel-B group.

[0061] In ADCS-ADL, the placebo group showed a significant decrease (worsening) of 9.27 ± 2.782 points at week 26 (p=0.0211; two-sided 95% CI -17.83 to -2.10) (Figure 4C). This change was significantly 6.95 ± 3.46 points at week 26 compared to the 20M lomesel-B group (p=0.0118; 95% CI 1.99 to 13.94). Similarly, this difference was also significant 6.96 ± 3.125 points at week 26 compared to the change in placebo in the combined lomesel-B group (p=0.0080; two-sided 95% CI 2.26 to 13.67). None of the lomesel-B groups showed a decrease from baseline.

[0062] In the ADRQL, the 100M lomesel-B group showed a significant improvement (increase) from baseline at week 2 (4.33 ± 5.88 points; p = 0.0449; 95% CI 0.12~8.54). The placebo group also showed significant increases at week 13 (2.21 ± 8.44 points; p = 0.0308; 95% CI 0.53~8.04) and week 26 (4.28 ± 4.49 points; p = 0.0225; 95% CI 0.97~8.83) (Figure 4D). The 20M lomesel-B group showed no significant change from baseline, and none of the lomesel-B groups showed a significant change compared to placebo.

[0063] Serum-based biomarkers: Post-treatment vascular biomarkers were significantly higher in the lomesel-B group compared to the placebo group. For VEGF, the placebo group showed a significant decrease up to 26 weeks compared to both the 20M lomesel-B (p<0.0128) and 100M lomesel-B (p<0.0012) groups (Figure 5A). Similarly, IL-4 was significantly reduced in the placebo group compared to both the 20M (p<0.0054) and 100M lomesel-B (p<0.0180) groups (Figure 5B). IL-6 was also significantly reduced in the placebo group compared to the 100M lomesel-B (p<0.0014) (Figure 5C). A significant increase in D-dimer levels was observed in the group receiving 100M of lomesel-B compared to placebo (Figure 5D), but no significant increase was observed in the group receiving 20M of lomesel-B compared to placebo.

[0064] Post-treatment anti-inflammatory biomarkers were significantly higher in the lomesel-B group compared to placebo. sIL-2Rα was significantly increased in the 100M lomesel-B group compared to placebo (p<0.0049) (Figure 5E). The 20M lomesel-B group significantly increased IL-10 (p<0.0349) (Figure 5F) and IL-12 (p<0.0015) (Figure 9E) compared to placebo.

[0065] Aβ 38 , Aβ 40 , and Aβ 42 Serum levels tended to be higher in the group treated with lomesel-B compared to placebo (Table 3).

[0066] [Table 3]

[0067] Hippocampal volume measurement: Brain volume measurements revealed a significant increase in left hippocampal volume at week 13 in the 100M Romecel-B group compared to the placebo group (p=0.0311) (Figure 6A). By week 26, this increase had diminished and was no longer statistically significant compared to placebo. The 20M Romecel-B group showed no significant difference compared to placebo. In contrast, no significant change was observed in the right hippocampus with or without Romecel-B compared to placebo (Figure 6B). In this analysis, hippocampal size was normalized to hippocampal sulcus volume to correct for differences in cranial size.

[0068] result: The key new findings from this placebo-controlled trial are that intravenous infusion of lomesel-B is safe and well-tolerated in patients with mild Alzheimer's disease (AD), potentially improving neurocognition and quality of life in treated patients and inducing biologically plausible changes in serum biomarkers. Furthermore, the trial revealed important insights into cellular dose and duration of effect, suggesting that lower doses may be more effective than higher doses. Taken together, the results of this trial pave the way for future large-scale clinical trials that will be enhanced to detect clinical efficacy endpoints.

[0069] This study is supported by preclinical results and is based on a robust pathophysiological therapeutic theory addressing the neuroinflammatory and vascular hypothesis of AD etiology. Considering the well-characterized anti-inflammatory and vascular effects of MSCs, we designed a placebo-controlled trial to evaluate them in mild AD.

[0070] Based on some evidence from pre-specified indicators, lomesel-B is recommended as a disease-modifying intervention for AD through improvements in all studied efficacy domains, including neurocognitive and neuropsychological, QOL and ADL, and biomarkers. In the clinical efficacy domain, the 20M lomesel-B group showed significant benefits compared to placebo in MMSE, patient QOL-AD, and ADRQL; however, this result is secondary and should be treated with caution. More importantly, none of the lomesel-B groups showed significant deterioration from baseline in any clinical assessment, which differs from the placebo group and further supports the safety of lomesel-B.

[0071] Regarding biomarkers, we detected significant changes in circulating biomarkers in two categories: vascular (VEGF, IL-4, IL-6) and anti-inflammatory (IL-4, IL-10, IL-12, and sIL-2Rα). These changes were primarily dose-dependent, with placebo leading to a decrease, a 100M dose showing the greatest significant increase, and a 20M dose being intermediate or similar to 100M.

[0072] Changes in vascular biomarkers are consistent with neurovascular improvements. VEGF, which showed significant changes, possesses neuroprotective and neuroregenerative effects and is actively associated with the increase in hippocampal volume observed in this study. IL-4 is a multifaceted cytokine that modulates vascular function, cell proliferation, and apoptosis, reduces pro-inflammatory profiles in various cell types including microglia, and induces BDNF production from astrocytes. IL-4 can also improve long-term potentiation (LTP) induced by Aβ inhibition by suppressing Aβ-induced upregulation of IL-1β through M1 microglia activation. IL-4 also facilitates the clearance of oligomeric Aβ peptides by increasing the expression of the Aβ-degrading enzyme CD10 in microglia. Furthermore, IL-4 can activate the M2 microglial phenotype, which promotes neurogenesis and oligodendrogenesis and is positively correlated with left hippocampal volume in patients with mild cognitive impairment. In vivo injection of IL-4 into the APP23 AD mouse model reduced Aβ levels and significantly improved memory impairment. Furthermore, IL-6 is a multifaceted cytokine that exhibits beneficial effects under exercise conditions, possesses angiogenesis-promoting and osteogenic activity, and can protect against glucose toxicity via VEGF signaling.

[0073] The increase in anti-inflammatory biomarkers in the lomesel-B treatment group is consistent with a reduction in systemic inflammation and neuroinflammation. Since neuroinflammation appears to be necessary for the development of dementia, the increase in the anti-inflammatory cytokine profile is consistent with improved clinical assessment. IL-10 has well-established anti-inflammatory properties. IL-12 has context-dependent anti-inflammatory and pro-inflammatory activity and induces IL-10 expression as part of its anti-inflammatory role. In the context of AD, IL-12 is significantly lower in the CSF of AD patients compared to normal subjects, and in this study, both IL-10 and IL-12 increased after lomesel-B treatment. At the peak of the anti-inflammatory roles of sIL-2Rα and IL-4, these results suggest a synergistic anti-inflammatory effect in response to lomesel-B treatment.

[0074] Circulating levels of Aβ peptide tended to be higher in the group treated with Lomesel-B compared to placebo. 42 This represents a moderate decrease in the preclinical / prodromal AD stage, and in AD, Aβ 40 and Aβ 42 This shows an even larger, significant decrease. The trend in Aβ observed in the lomesel-B group is consistent with the improvement in cognitive status observed in patients.

[0075] Finally, neurogenesis is significantly reduced in adults with Alzheimer's disease (AD). The increase in hippocampal volume appears to be consistent with the increased neurogenesis and improvements in other efficacy domains in these patients.

[0076] References 1. Sheppard O, Coleman M. Alzheimer's Disease: Etiology, Neuropathology and Pathogenesis. In: Huang X, editor. Alzheimer's Disease: Drug Discovery. Brisbane (AU)2020. p. 6. 2. Lue LF, Brachova L, Civin WH, Rogers J. Inflammation, A beta deposition, and neurofibrillary tangle formation as correlates of Alzheimer's disease neurodegeneration. J Neuropathol Exp Neurol. 1996;55(10):1083-8. Epub 1996 / 10 / 01. PubMed PMID: 8858005. 3. Pimplikar SW. Neuroinflammation in Alzheimer’s disease: from pathogenesis to a therapeutic target. J Clin Immunol. 2014;34 Suppl 1:S64-9. Epub 2014 / 04 / 09. doi: 10.1007 / s10875-014-0032-5. PubMed PMID: 24711006. 4. Heneka MT, Carson MJ, El Khoury J, Landreth GE, Brosseron F, Feinstein DL, et al. Neuroinflammation in Alzheimer’s disease. Lancet Neurol. 2015;14(4):388-405. Epub 2015 / 03 / 21. doi: 10.1016 / S1474-4422(15)70016-5. PubMed PMID: 25792098. 5. Monson NL, Ireland SJ, Ligocki AJ, Chen D, Rounds WH, Li M, et al. Elevated CNS inflammation in patients with preclinical Alzheimer’s disease. J Cereb Blood Flow Metab. 2014;34(1):30-3. doi: 10.1038 / jcbfm.2013.183. PubMed PMID: 24149932; PubMed Central PMCID: PMC3887357. 6. Steinman J, Sun HS, Feng ZP. Microvascular Alterations in Alzheimer’s Disease. Front Cell Neurosci. 2020;14:618986. doi: 10.3389 / fncel.2020.618986. PubMed PMID: 33536876; PubMed Central PMCID: PMCPMC7849053. 7. Marques F, Sousa JC, Sousa N, Palha JA. Blood-brain-barriers in aging and in Alzheimer’s disease. Mol Neurodegener. 2013;8:38. doi: 10.1186 / 1750-1326-8-38. PubMed PMID: 24148264; PubMed Central PMCID: PMCPMC4015275. 8. Zhao Z, Nelson AR, Betsholtz C, Zlokovic BV. Establishment and Dysfunction of the Blood-Brain Barrier. Cell. 2015;163(5):1064-78. doi: 10.1016 / j.cell.2015.10.067. PubMed PMID: 26590417; PubMed Central PMCID: PMCPMC4655822. 9. Tarasoff-Conway JM, Carare RO, Osorio RS, Glodzik L, Butler T, Fieremans E, et al. Clearance systems in the brain-implications for Alzheimer disease. Nat Rev Neurol. 2015;11(8):457-70. doi: 10.1038 / nrneurol.2015.119. PubMed PMID: 26195256; PubMed Central PMCID: PMCPMC4694579. 10. Erickson MA, Banks WA. Blood-brain barrier dysfunction as a cause and consequence of Alzheimer’s disease. J Cereb Blood Flow Metab. 2013;33(10):1500-13. doi: 10.1038 / jcbfm.2013.135. PubMed PMID: 23921899; PubMed Central PMCID: PMCPMC3790938. 11. Hayashi S, Sato N, Yamamoto A, Ikegame Y, Nakashima S, Ogihara T, et al. Alzheimer disease-associated peptide, amyloid beta40, inhibits vascular regeneration with induction of endothelial autophagy. Arterioscler Thromb Vasc Biol. 2009;29(11):1909-15. Epub 2009 / 10 / 10. doi: 10.1161 / ATVBAHA.109.188516. PubMed PMID: 19815818. 12. Klohs J. An Integrated View on Vascular Dysfunction in Alzheimer’s Disease. Neurodegener Dis. 2019;19(3-4):109-27. doi: 10.1159 / 000505625. PubMed PMID: 32062666. 13. Oliva AA, McClain-Moss L, Pena A, Drouillard A, Hare JM. Allogeneic mesenchymal stem cell therapy: A regenerative medicine approach to geroscience. Aging Med (Milton). 2019;2(3):142-6. doi: 10.1002 / agm2.12079. PubMed PMID: 31667462; PubMed Central PMCID: PMCPMC6820701. 14. Pittenger MF, Discher DE, Peault BM, Phinney DG, Hare JM, Caplan AI. Mesenchymal stem cell perspective: cell biology to clinical progress. NPJ Regen Med. 2019;4:22. doi: 10.1038 / s41536-019-0083-6. PubMed PMID: 31815001; PubMed Central PMCID: PMCPMC6889290. 15. Eggenhofer E, Luk F, Dahlke MH, Hoogduijn MJ. The life and fate of mesenchymal stem cells. Front Immunol. 2014;5:148. doi: 10.3389 / fimmu.2014.00148. PubMed PMID: 24904568; PubMed Central PMCID: PMC4032901. 16. Lalu MM, McIntyre L, Pugliese C, Fergusson D, Winston BW, Marshall JC, et al. Safety of cell therapy with mesenchymal stromal cells (SafeCell): a systematic review and meta-analysis of clinical trials. PloS one. 2012;7(10):e47559. doi: 10.1371 / journal.pone.0047559. PubMed PMID: 23133515; PubMed Central PMCID: PMCPMC3485008. 17. Karantalis V, Schulman IH, Balkan W, Hare JM. Allogeneic cell therapy: a new paradigm in therapeutics. Circ Res. 2015;116(1):12-5. doi: 10.1161 / CIRCRESAHA.114.305495. PubMed PMID: 25552688; PubMed Central PMCID: PMCPMC4411634. 18. Neves AF, Camargo C, Premer C, Hare JM, Baumel BS, Pinto M. Intravenous administration of mesenchymal stem cells reduces Tau phosphorylation and inflammation in the 3xTg-AD mouse model of Alzheimer’s disease. Experimental neurology. 2021:113706. doi: 10.1016 / j.expneurol.2021.113706. PubMed PMID: 33757765. 19. Kim S, Chang KA, Kim J, Park HG, Ra JC, Kim HS, et al. The preventive and therapeutic effects of intravenous human adipose-derived stem cells in Alzheimer’s disease mice. PloS one. 2012;7(9):e45757. doi: 10.1371 / journal.pone.0045757. PubMed PMID: 23049854; PubMed Central PMCID: PMC3458942. 20. Shin JY, Park HJ, Kim HN, Oh SH, Bae JS, Ha HJ, et al. Mesenchymal stem cells enhance autophagy and increase beta-amyloid clearance in Alzheimer disease models. Autophagy. 2014;10(1):32-44. doi: 10.4161 / auto.26508. PubMed PMID: 24149893. 21. Lee JK, Schuchman EH, Jin HK, Bae JS. Soluble CCL5 derived from bone marrow-derived mesenchymal stem cells and activated by amyloid beta ameliorates Alzheimer’s disease in mice by recruiting bone marrow-induced microglia immune responses. Stem Cells. 2012;30(7):1544-55. doi: 10.1002 / stem.1125. PubMed PMID: 22570192. 22. Bae JS, Jin HK, Lee JK, Richardson JC, Carter JE. Bone marrow-derived mesenchymal stem cells contribute to the reduction of amyloid-beta deposits and the improvement of synaptic transmission in a mouse model of pre-dementia Alzheimer’s disease. Curr Alzheimer Res. 2013;10(5):524-31. doi: 10.2174 / 15672050113109990027. PubMed PMID: 23036020.

Claims

1. A method for alleviating symptoms of Alzheimer's disease (AD) in a subject requiring it, comprising the step of administering a composition containing a therapeutically effective amount of allogenic mesenchymal stem cells (MSCs) to the subject.

2. A method for treating Alzheimer's disease (AD) or inhibiting the progression of AD, comprising the step of administering a composition containing a therapeutically effective amount of allogenic mesenchymal stem cells (MSCs) to the subject.

3. The method according to claim 1 or 2, further comprising the step of measuring the concentration of one or more biomarkers in a subject suffering from symptoms of AD before and after administration of the composition containing allogenic MSCs.

4. The method according to any one of claims 1 to 3, further comprising the step of measuring the cognitive function of a subject suffering from symptoms of AD before and after administration of the composition containing allogenic MSCs.

5. The method according to claim 3 or 4, wherein the biomarker comprises a cytokine selected from the group consisting of IL-4, IL-6, IL-8, IL-10, IL-12p70, IL-17, sIL-2Rα, or a combination thereof.

6. The method according to claim 5, wherein the concentration of the cytokine increases in the serum, plasma, cerebrospinal fluid, or blood of a subject requiring the treatment after administration of the composition containing a therapeutically effective amount of allogenic MSCs.

7. The method according to claim 6, wherein the increase in the concentration of the cytokine is 0.5% to 10%, 5% to 10%, 10% to 50%, or more than 50%.

8. The method according to any one of claims 3 to 7, wherein the biomarker further comprises a neuron-related molecule or peptide selected from the group consisting of tau, phosphotau, Aβ-38, Aβ-40, Aβ-42, NFL, or a combination thereof.

9. The method according to claim 8, wherein the concentration of Aβ-38, Aβ-40, or Aβ-42 increases in the serum, plasma, or blood of a subject requiring the treatment after administration of the composition containing a therapeutically effective amount of allogenic MSCs.

10. The method according to claim 9, wherein the concentration of Aβ-38, Aβ-40, or Aβ-42 increases by 0.5% to 10%, 5% to 10%, 10% to 50%, or more than 50%.

11. The method according to claim 8, wherein the concentration of tau, phosphotau, or NFL decreases in the serum, plasma, cerebrospinal fluid, or blood of a subject requiring the treatment after administration of the composition comprising a therapeutically effective amount of allogenic MSCs.

12. The method according to claim 11, wherein the concentration of tau, phosphotau, or NFL decreases by 0.5% to 10%, 5% to 10%, 10% to 50%, or more than 50%.

13. The method according to any one of claims 3 to 12, wherein the biomarker further comprises an inflammation signaling molecule such as pro-BNP, TNF-α, or a combination thereof.

14. The method according to claim 13, wherein the concentration of pro-BNP or TNF-α decreases in the serum, plasma, cerebrospinal fluid or blood of a subject requiring it after administration of the composition comprising a therapeutically effective amount of allogenic MSC.

15. The method according to claim 14, wherein the concentration of the pro-BNP or TNF-α decreases by 0.5% to 10%, 5% to 10%, 10% to 50%, or more than 50%.

16. The method according to any one of claims 3 to 15, wherein the biomarker further comprises VEGF.

17. The method according to claim 16, wherein the concentration of VEGF increases in the serum, plasma, cerebrospinal fluid, or blood of a subject requiring it after administration of the composition containing a therapeutically effective amount of allogenic MSCs.

18. The method according to claim 17, wherein the concentration of VEGF decreases to 0.5% to 10%, 5% to 10%, 10% to 50%, or more than 50%.

19. The method according to any one of claims 3 to 18, further comprising the step of determining a change in the size of a region in the target brain after administration of the composition comprising allogenic MSCs.

20. The method according to claim 19, wherein the region in the target brain whose size changes after administration of the composition is selected from the group consisting of the amygdala, cortical nuclei, hippocampus, hippocampal microregions, and / or corticoamygdal junction.

21. The method according to any one of claims 3 to 20, further comprising the step of determining whether a change occurs in the target corticoamygdal junction after administration of the composition containing allogenic MSCs.

22. The above composition is 20 × 10 ６ The method according to any one of claims 3 to 21, comprising MSCs.

23. The above composition is 100 × 10 ６ The method according to any one of claims 3 to 21, comprising MSCs.

24. The method according to any one of claims 3 to 23, further comprising the step of examining the cerebrospinal fluid of the subject before and after administration of the composition containing allogenic MSCs.

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