Treating inflammation

Abstract

The present disclosure relates to a method of treating neuroinflammation, which in one aspect provides administering to a subject with neuroinflammation an effective amount of at least four herbal ingredients: Polygalae (Primarinus officinalis), Astragali (Astragalus membranaceus), Ligusticum Chuanxiong (Cnidium rhizome), and Angelica sinensis (Angelica sinensis), their roots or rhizomes, or extracts thereof.

Description

[Technical Field]

[0001] The present disclosure relates to methods for treating, controlling, preventing, and / or reducing the risk of inflammation in the central nervous system (CNS) by administering to a subject an herbal composition comprising a combination of traditional Chinese herbal medicines or extracts thereof. [Background technology]

[0002] Neuroinflammation is inflammation of nervous tissue, such as the brain and spinal cord. Neuroinflammation is an innate immune system response of the CNS that is triggered after insults caused by, for example, injury, infection, exposure to toxins, neurodegenerative disease, aging (e.g., "inflammage"), or resulting from chemical, mechanical, or surgical interventions to treat underlying conditions or diseases.

[0003] Neuroinflammation can be mediated by the production of reactive oxygen species, chemokines, and cytokines. These mediators are produced by CNS glia, including microglia and astrocytes, as well as endothelial cells and peripherally derived immune cells. Acting similarly to macrophages during systemic inflammation, microglial cells are thought to be a key cell type involved in neuroinflammation. In response to injury, microglia have the ability to transition to different functional states, altering their proliferation, morphology (i.e., shortened processes), phagocytic activity (removal of foreign organisms and cellular debris), and antigen presentation. However, in addition to these beneficial processes in response to injury, microglial cells also release pro-inflammatory factors, such as cytokines, chemokines, and matrix metalloproteinases (MMPs), which can impair blood-brain barrier (BBB) ​​function. Overall, the multiphasic (beneficial and detrimental) roles of neuroinflammation are time-dependent, but prolonged activation of microglia causes damage to brain tissue and the BBB.

[0004] There is a lack of effective and safe therapeutic agents that can modulate neuroinflammation to limit or prevent its adverse effects on brain tissue and the BBB. Therefore, new therapeutic methods for modulating neuroinflammation are urgently needed. Summary of the Invention [Problem to be solved by the invention]

[0005] The present disclosure provides a combination of traditional Chinese herbal medicines or extracts thereof, which combination reduces microglial activation and secretion of inflammatory cytokines and other mediators of neuroinflammation after injury, and is therefore particularly effective in preventing and / or controlling neuroinflammation, treating, reducing, or preventing symptoms associated with microglial overactivation, and mitigating the immune, physiological, biochemical, and psychological consequences of neuroinflammation, including damage to brain tissue and the BBB. [Means for solving the problem]

[0006] Thus, according to a first aspect, there is provided a method of treating neuroinflammation in a subject in need thereof, comprising administering to the subject an effective amount of an herbal composition comprising at least four herbal ingredients: Polygalae (Primarinus officinalis), Astragali (Astragalus membranaceus), Ligusticum chuanxiong (Cnidium rhizome), and Angelica sinensis (Angelica sinensis), their roots or rhizomes, or extracts thereof. As used in this aspect, as well as the following aspects and embodiments, "treating" has the meaning defined in the subsequent section entitled "Glossary," unless specifically stated below to have a different meaning.

[0007] In one embodiment of the above aspect, the herbal composition comprising four herbal ingredients, Polygalae (Princess bush clover), Astragali (Astragali), Ligusticum Chuanxiong, and Angelica sinensis (Angelica sinensis), their roots or rhizomes, or extracts thereof, further comprises at least one herbal composition selected from the group consisting of Salviae Miltiorrhizae (Red sage), Paeoniae Rubra (Red peony), Carthamus Tinctorius (Safflower) flower, Semen Persicae (Peach (Prunus Persica) seed), and Acori Tatarinowii (Stone iris), their roots or rhizomes, or extracts thereof.

[0008] In a further aspect, there is provided a method of treating neuroinflammation in a subject in need thereof, comprising administering to the subject an effective amount of an herbal composition consisting essentially of at least four herbal components: Polygalae (Primarinus officinalis), Astragali (Astragalus membranaceus), Ligusticum Chuanxiong, and Angelica sinensis (Angelica sinensis), their roots or rhizomes, or extracts thereof.

[0009] In another aspect, there is provided a method of treating neuroinflammation in a subject in need thereof, comprising administering to the subject an effective amount of an herbal composition consisting essentially of at least nine herbal ingredients: Polygalae (Princess bush clover), Astragali (Astragali root), Ligusticum Chuanxiong (Cnidium rhizome), Angelica sinensis (Angelica sinensis), Salviae Miltiorrhizae (Red sage), Paeoniae Rubra (Red peony), Carthamus tinctorius (Safflower) flower, Semen persicae (Peach (Prunus persica) seed), and Acori tatarinowii (Stone iris), their roots or rhizomes, or extracts thereof.

[0010] In one embodiment of any of the above aspects, the herbal composition is a pharmaceutical composition. The pharmaceutical composition may also include one or more pharmaceutically acceptable carriers or excipients. In one embodiment of any of the above aspects, the neuroinflammation is associated with an injury such as a spinal cord injury, stroke, traumatic brain injury, subarachnoid hemorrhage, subdural hemorrhage, or epidural hemorrhage.

[0011] In one embodiment of any of the above aspects, the neuroinflammation is associated with an infection, such as an infection of the brain or central nervous system (e.g., encephalitis or bacterial, viral, or fungal meningitis), or a severe systemic infection, such as sepsis or an abscess. Examples of viral infections include varicella-zoster virus (VZV), herpes simplex virus (HSV-1 or HSV-2), SARS-CoV-2 or a variant thereof, Epstein-Barr virus, hepatitis B virus, hepatitis C virus, human immunodeficiency virus type 1 (HIV-1), human papillomavirus (HPV), human T-cell lymphotropic virus type 1 (HTLV-1), and Kaposi's sarcoma herpesvirus (KSHV). Examples of fungi that can cause neuroinflammation include Cryptococcus neoformans, Candida albicans, Histoplasma capsulatum, Coccidioides immitis, Paracoccidioides brasiliensis, Aspergillus spp., and zygomycetes. Examples of bacteria that can cause neuroinflammation include Listeria monocytogenes, Borrelia burgdorferi, Neisseria meningitidis, Streptococcus pneumoniae, Staphylococcus aureus, Haemophilus influenzae, Mycobacterium, and Brucella species. Neuroinflammation can also be associated with parasitic infections.

[0012] In one embodiment of any of the above aspects, the neuroinflammation is associated with exposure to a toxin. In one embodiment of any of the above aspects, neuroinflammation is associated with neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, Huntington's disease, frontotemporal dementia, amyotrophic lateral sclerosis, multiple sclerosis, 10-12 primary tauopathies, synucleinopathies (i.e., dementia with Lewy bodies and multiple system atrophy), polyglutamine diseases including spinocerebellar ataxias, prion diseases, traumatic brain injury, chronic traumatic encephalopathy, stroke, and spinal cord injury.

[0013] In one embodiment of any of the above aspects, the neuroinflammation is associated with a neuropsychiatric condition such as depression, stress, post-traumatic stress disorder, schizophrenia, autism spectrum disorder, or epilepsy.

[0014] In one embodiment of any of the above aspects, the neuroinflammation is associated with aging (e.g., "inflammagenesis"). Other disorders that can cause neuroinflammation include, for example, postoperative cognitive dysfunction, ischemia, brain tumors, chronic pain, headache, migraine, trigeminal neuralgia, cardiovascular disease and associated risk factors (e.g., due to obesity, hypertension, diabetes, hyperlipidemia, smoking, etc.), chronic peripheral inflammatory conditions (e.g., rheumatoid arthritis, inflammatory bowel disease, psoriasis, etc.), perturbation of the gut microbiota (i.e., dysbiosis), and herpes keratitis.

[0015] In one embodiment of any of the above aspects, the neuroinflammation is associated with chemical intervention to treat the underlying disease or disorder. One example of such chemical intervention is the use of TNF inhibitors to treat autoimmune diseases such as rheumatoid arthritis.

[0016] In one embodiment of any of the above aspects, the neuroinflammation is associated with a mechanical intervention to treat an underlying disease or disorder. In one embodiment of any of the above aspects, the neuroinflammation is associated with surgical intervention to treat the underlying disease or disorder. Examples of surgical intervention include carotid endarterectomy, carotid angioplasty with or without stenting, vertebral angioplasty with or without stenting, intracranial angioplasty with or without stenting, aneurysm coiling, occlusion of vascular malformations, repair of intracranial fistulas, thrombectomy, extracranial-intracranial bypass, open-heart surgery, cardiac surgery requiring bypass surgery, coronary artery bypass grafting, coronary angioplasty with or without stenting, valve surgery / valvuloplasty, valvotomy, closure of patent foramen ovale, and surgery on the great vessels of the heart.

[0017] In one embodiment of any of the above aspects, there is provided administration of an effective amount of the herbal composition that is from about 1 mg / kg to about 100 mg / kg. In one embodiment of any of the above aspects, there is provided a method comprising administering a second agent for treating the underlying disease or disorder together with the aforementioned herbal composition.

[0018] In one embodiment of any of the above aspects, there is provided administration of an effective amount of the herbal composition to control neuroinflammation and promote recovery after a brain lesion. In one embodiment of any of the above aspects, there is provided administration of an effective amount of the herbal composition to protect the brain from the damaging effects of neuroinflammation and promote recovery after a brain lesion.

[0019] In one embodiment of any of the above aspects, there is provided administration of an effective amount of the herbal composition to modulate the inflammatory cascade to promote neurovascular repair. In one embodiment of any of the above aspects, there is provided administration of an effective amount of the herbal composition to provide a barrier against the damaging effects of neuroinflammation to promote recovery after a brain lesion.

[0020] In one embodiment of any of the above aspects, there is provided administration of an effective amount of the herbal composition to strengthen the blood-brain barrier against the damaging effects of neuroinflammation and promote recovery after brain lesions. In certain embodiments of any of the above aspects and embodiments, neuroinflammation is treated or alleviated in a subject with an underlying disease. In one specific embodiment, the underlying disease is diabetes mellitus.

[0021] The accompanying drawings are included to illustrate specific embodiments of the present disclosure and related technology. The drawings provide a further understanding of the present disclosure and are incorporated in and constitute a part of this disclosure. It should be understood that the drawings illustrate implementations of the present disclosure and, together with the remainder of the disclosure, explain the principles of the present disclosure. The drawings illustrate embodiments of the present disclosure, and it will be apparent to those skilled in the art that other embodiments can be readily understood from the drawings described herein. [Brief explanation of the drawings]

[0022] [Figure 1] FIG. 1 shows the time course of the experimental protocol for in vitro inflammation testing. [Figure 2] FIG. 1 shows the time course of the experimental protocol for in vivo studies. [Figure 3] 1 is a table showing the sequences of the qPCR (quantitative polymerase chain reaction) primers used. [Figure 4] FIG. 1 shows the effect of MLC1501 (defined below) on the relative expression of interleukin-1β (IL-1β), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), NOD-, LRR-, and pyrin domain-containing protein 3 (NLRP3), and inducible nitric oxide synthase (iNOS) genes after stimulation of immortalized mouse microglial cell line (BV2) cells with lipopolysaccharide (LPS). [Figure 5] FIG. 1 shows the dose-response effect of MLC1501 on the LPS-induced increase in the relative expression of IL-1β, IL-6, and TNF-α on BV2 cells. [Figure 6] FIG. 1 shows the effect of MLC1501 on the relative expression of IL-1β and IL-6 after middle cerebral artery occlusion (MCAO)-induced cerebral ischemia. DETAILED DESCRIPTION OF THE INVENTION

[0023] Glossary This section is intended to provide guidance on the interpretation of the words and phrases listed below (and their grammatical variations, where appropriate).

[0024] Unless otherwise noted, the terms "comprising" and "comprise," and grammatical variations thereof, are intended to express "open" or "inclusive" language, including the recited elements and allowing for the inclusion of additional, unrecited elements.

[0025] As used herein, the term "about" when used in reference to a numerical value means, for example, about ±30%, about ±20%, about ±10%, about ±5%, or about ±1% of the numerical value. Where appropriate, the word "about" may be omitted from the definitions of this disclosure.

[0026] The words "a," "an," and "the" are used to describe elements and components of the present invention. This is merely for convenience and to convey a general sense of the invention. This description is to be construed as including one or at least one, and singular forms also include plural forms, unless the context clearly dictates otherwise. Thus, for example, the term "agent" includes reference to a single agent as well as multiple agents (including agent mixtures). It should also be noted that the term "or" is normally used in its sense to include "and / or" unless the context clearly dictates otherwise.

[0027] As used herein, the term "inflammaging" refers to the age-associated increase in pro-inflammatory markers in blood and / or tissues that may contribute to the pathogenesis of age-associated diseases. As used herein, the term "in vivo" includes the use of a living whole organism. This is in contrast to the term "in vitro," which does not use a whole organism. The term "in vitro" is understood to include "ex vivo" uses, which include, inter alia, the use of cells, tissues, etc. that do not form part of a whole organism (e.g., cells or tissues from cell or tissue culture, biopsies, dead organisms, etc.). A further non-limiting example of "in vitro" relates to the use of cell extracts or lysates.

[0028] The terms "patient" and "subject" are used interchangeably herein and include reference to any human or non-human animal (preferably a mammalian subject) that one desires to treat using the present invention. However, it will be understood that "patient" or "subject" does not imply the presence of a condition. The term "mammal," as used herein, refers to any animal belonging to the class Mammalia, including, but not limited to, humans and non-human primates, such as apes and monkeys, including chimpanzees; livestock, such as cows, sheep, pigs, goats, and horses; pet / companion animals, such as dogs and cats; and laboratory animals (e.g., rabbits and rodents, such as mice, rats, and guinea pigs). Preferably, the mammal is a human.

[0029] The terms "treatment" and "treating" include any use that in any way corrects a disease state or symptom, prevents the establishment of a disease, or prevents, hinders, slows, or reverses the progression of a disease or other undesirable symptoms. Thus, "treatment" and "treating" include prophylactic and therapeutic treatments. In the context of "methods of treating neuroinflammation" herein, "treating" encompasses modulating neuroinflammation to limit or prevent its adverse effects on brain tissue and the BBB, while limiting its impact on its beneficial effects.

[0030] Throughout this disclosure, certain embodiments may be disclosed in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the disclosed ranges. Thus, the description of a range should be considered to have specifically disclosed not only each individual numerical value within that range but also all possible subranges. For example, a description of a range such as 1 to 6 should be considered to have specifically disclosed subranges such as 1 to 3, 1 to 4, 1 to 5, 2 to 4, 2 to 6, 3 to 6, etc., as well as each individual numerical value within that range, e.g., 1, 2, 3, 4, 5, and 6. This applies regardless of the breadth of the range.

[0031] Unless the context clearly indicates otherwise, the terms "disease," "disorder," and "condition" may be used interchangeably herein. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains.

[0032] Detailed embodiments and implementations of the claimed subject matter are disclosed in detail herein, along with technical matters, structural features, objectives achieved, and advantages, with reference to the accompanying drawings. It should be understood that the disclosed embodiments and implementations are merely exemplary of the claimed subject matter, which may be embodied in various forms. However, the present disclosure may be embodied in many different forms and should not be construed as being limited to the exemplary embodiments and implementations described herein. Rather, these exemplary embodiments and implementations are provided so that the description of the present disclosure will be thorough and complete, and the scope of the disclosure will be fully conveyed to those skilled in the art. In particular, the terms used in the embodiments of the present invention are intended to describe the purpose of particular embodiments and not to limit the present disclosure. In the following description, details of well-known features and techniques may be omitted to avoid unnecessarily obscuring the presented embodiments and implementations.

[0033] The configurations described in the following description are non-limiting examples that may be modified and are cited merely to illustrate at least one embodiment and are not intended to limit the scope thereof.

[0034] Microglia are the main resident macrophages in the central nervous system, accounting for 5–12% of brain cells. Microglia act as cellular effectors and thus play a key role in the immune response of the central nervous system. Therefore, microglia are important in the body's defense and tissue repair processes under physiological conditions. However, pro-inflammatory immune responses driven by microglia are also important contributors to the pathogenesis of several brain diseases. Exacerbated activation of microglia, for example, by LPS or ischemia, can induce harmful inflammatory responses by resulting in the release of chemokines and pro-inflammatory cytokines such as IL-1β, IL-6, and TNF-α. This release of cytokines and chemokines can occur through various intermediates.

[0035] One key step in the inflammatory response is the activation of inflammasomes, intracellular protein complexes containing pattern recognition receptors and other inflammatory molecules. IL-1β secretion is known to be directly mediated by the formation of inflammasomes containing NLRP3.

[0036] Another mediator of pro-inflammatory cytokine and chemokine secretion is iNOS, which can be induced by both endotoxins (such as LPS) and cytokines. Its induction leads to the production of nitric oxide (NO), a free radical associated with microglial cytotoxic function, apoptosis, and oxidative stress.

[0037] In addition to causing neuronal cell death, these cytokines and pro-inflammatory mediators promote damage to the blood-brain barrier and thus leukocyte infiltration into the brain. Certain herbal compositions are identified herein that limit the release of pro-inflammatory cytokines, including cytokines IL-1β, IL-6, and TNF-α. The herbal compositions herein also limit the formation of inflammasomes, including NLRP3 and iNOS. Thus, the herbal compositions are particularly useful for treating inflammation involving these cytokines and inflammasomes, including treating neuroinflammation.

[0038] Accordingly, in one aspect of the present disclosure, there is provided a method of treating neuroinflammation in a subject in need thereof, comprising administering to the subject an effective amount of an herbal composition comprising at least four herbal ingredients, Radix Polygalae (Primula Root), Radix Astragali (Astragalus Root), Rhizome Ligusticum Chuanxiong (Cnidium Rhizome), and Radix Angelica sinensis (Angelica Root), or extracts thereof.

[0039] In one embodiment of the above aspect, the herbal composition comprises four herbal ingredients, Polygala Root (Radix Polygalae) (Root of Japanese Bush Clovers), Astragalus Root (Radix Astragali) (Root of Astragali Root), Cnidium Rhizome (Rhizome Ligusticum Chuanxiong), and Angelica Root (Radix Angelica sinensis) (Root of Chinese Angelica), and further comprises at least one herbal ingredient selected from the group consisting of Salvia miltiorrhizae (Red Sage Root), Radix Paeoniae Rubra (Red Peony Root), Carthamus Tinctorius (Safflower) Flower, Peach Seed (Semen Persicae) (Peach (Prunus Persica) Seed), and Rhizome Acori Tatarinowii (Rhizome of Japanese Calamus) or extracts thereof.

[0040] In a further aspect, there is provided a method of treating neuroinflammation in a subject in need thereof, comprising administering to the subject an effective amount of an herbal composition consisting essentially of at least four herbal ingredients, Radix Polygalae (Primula Root), Radix Astragali (Astragalus Root), Rhizome Ligusticum Chuanxiong (Cnidium Rhizome), and Radix Angelica sinensis (Angelica Root), or extracts thereof.

[0041] In another embodiment, at least nine herbal ingredients, Polygala Root (Radix Polygalae) (Hibaki Root), Astragalus Root (Radix Astragali) (Astragalus Root), Cnidium Rhizome (Rhizome Ligusticum Chuanxiong) and Angelica Root (Radix Angelica sinensis) (Angelica Root), Salvia Root and Rhizome (Radix et Rhizome Salviae Miltiorrhizae) (Red Sage Root), Radix Paeoniae Rubra (Red Peony Root), Carthamus Tinctorius (Safflower) Flower, Peach Seed (Semen Persicae) (Peach (Prunus Persica) Seed), and Rhizome Acori In one embodiment, a method of treating neuroinflammation in a subject in need thereof is provided, comprising administering to the subject an effective amount of an herbal composition consisting essentially of Acanthus tatarinowii (the rhizome of Calamus sieboldii) or an extract thereof.

[0042] In one embodiment of any of the above aspects, neuroinflammation is treated or alleviated in a subject with an underlying disease, such as cardiovascular disease, diabetes, obesity, or atherosclerosis. In one specific embodiment, a subject with an underlying disease, such as cardiovascular disease, diabetes, obesity, or atherosclerosis, is treated with the above-described herbal composition to reduce the risk of damaging effects on the brain caused by neuroinflammation. In one specific embodiment, the underlying disease is diabetes mellitus. In one specific embodiment, a diabetic subject is treated with the above-described herbal composition to improve post-stroke recovery after an ischemic stroke.

[0043] In one embodiment of any of the above aspects, the herbal composition is a pharmaceutical composition. The pharmaceutical composition may also include one or more pharmaceutically acceptable carriers or excipients. In one embodiment of any of the above aspects, the herbal composition comprises an extract of the herbal ingredients described above.

[0044] In one embodiment of any of the above aspects, the herbal composition comprises 5 times the weight to weight ratio of Radix Astragali or an extract thereof compared to the weight of each of the other herbal ingredients or extracts thereof present.

[0045] In one embodiment of any of the above aspects, the pharmaceutical composition is MLC901. In one embodiment of any of the above aspects, the pharmaceutical composition is MLC1501. In one embodiment of any of the above aspects, the neuroinflammation is the result of an injury such as an injury selected from a spinal cord injury, stroke, traumatic brain injury, subarachnoid hemorrhage, subdural hemorrhage, or epidural hemorrhage.

[0046] In certain embodiments of any of the above aspects, the neuroinflammation occurs after an ischemic stroke. In this embodiment, treatment of a subject with the herbal compositions described herein, including treatment with MLC901 or treatment with MLC1501, provides: 1. An increased rate of functional independence / autonomy restoration between about 3 months and about 24 months after the initial administration of the herbal composition. In one aspect, the restoration of functional independence / autonomy results from the control, modulation, protection, and / or strengthening of the blood-brain barrier against the damaging effects of neuroinflammation; 2. A reduction in time to achieve restoration of functional independence / autonomy of approximately 18 months. In one aspect, restoration of functional independence / autonomy results from control, modulation, protection, and / or strengthening of the blood-brain barrier against the damaging effects of neuroinflammation; 3. Acceleration in achieving restoration of functional independence / autonomy. In one aspect, restoration of functional independence / autonomy results from the control, modulation, protection, and / or strengthening of the blood-brain barrier against the damaging effects of neuroinflammation; 4. Reduced length of hospital stay. In one aspect, reduced time spent in the hospital is a result of controlling, modulating, protecting, and / or strengthening the blood-brain barrier against the damaging effects of neuroinflammation; 5. Reduction in time and / or costs associated with regaining functional independence / autonomy. In one aspect, regaining functional independence / autonomy results from controlling, regulating, protecting, and / or strengthening the blood-brain barrier against the damaging effects of neuroinflammation; and / or 6. Improved quality of life. In one aspect, the improvement results from controlling, regulating, protecting, and / or strengthening the blood-brain barrier against the damaging effects of neuroinflammation.

[0047] In certain embodiments of any of the above aspects, neuroinflammation occurs after a trauma-induced neurodegenerative disease, such as traumatic brain injury (TBI) or spinal cord injury. In this embodiment, treatment of a subject with the herbal compositions described herein, including treatment with MLC901 or treatment with MLC1501, provides a reduction in the severity of symptoms following injury. In certain embodiments, the reduction in the severity of symptoms following injury is achieved by regulating, modulating, protecting, and / or strengthening the blood-brain barrier against the damaging effects of neuroinflammation. Following TBI, treatment of a subject with the herbal compositions described herein, including treatment with MLC901 or treatment with MLC1501, may also provide improved recovery of cognitive domains, such as complex attention and / or executive function, and / or may have a positive effect on anxiety or depression symptoms and / or quality of life. In certain embodiments, recovery of cognitive domains is achieved by regulating, modulating, protecting, and / or strengthening the blood-brain barrier against the damaging effects of neuroinflammation.

[0048] In certain embodiments of any of the above aspects, treatment of a subject with the herbal compositions described herein, including treatment with MLC901 or treatment with MLC1501, ameliorates mood disorders associated with neurodegenerative diseases. In certain embodiments, amelioration of mood disorders is achieved by regulating, modulating, protecting, and / or strengthening the blood-brain barrier against the damaging effects of neuroinflammation.

[0049] In certain embodiments of any of the above aspects, the neuroinflammation is due to a subject suffering from dementia or Alzheimer's disease. In this embodiment, treatment of a subject with the herbal compositions described herein, including treatment with MLC901 or treatment with MLC1501, provides: 1. Improved recovery of cognitive function (e.g., memory, habits, and / or language) within about 3 months to about 48 months after initial administration of the herbal composition. In one aspect, the improved recovery results from regulating, modulating, protecting, and / or strengthening the blood-brain barrier against the damaging effects of neuroinflammation; 2. Slowing of disease progression. In one aspect, the slowing of disease progression results from regulating, modulating, protecting, and / or strengthening the blood-brain barrier against the damaging effects of neuroinflammation; and / or 3. Disease-Modifying Effects. In one aspect, the disease-modifying effects result from regulating, modulating, protecting, and / or strengthening the blood-brain barrier against the damaging effects of neuroinflammation.

[0050] In one embodiment of any of the above aspects, the neuroinflammation is the result of an injury, e.g., an infection, e.g., of the brain or central nervous system (e.g., encephalitis or bacterial, viral, or fungal meningitis), or a severe infection, e.g., sepsis or an abscess. Examples of viral infections include varicella-zoster virus (VZV), herpes simplex virus (HSV-1 or HSV-2), SARS-CoV-2 or a variant thereof, Epstein-Barr virus, hepatitis B virus, hepatitis C virus, human immunodeficiency virus type 1 (HIV-1), human papillomavirus (HPV), human T-cell lymphotropic virus type 1 (HTLV-1), and Kaposi's sarcoma herpesvirus (KSHV). Examples of fungi that can cause neuroinflammation include Cryptococcus neoformans, Candida albicans, Histoplasma capsulatum, Coccidioides immitis, Paracoccidioides brasiliensis, Aspergillus spp., and zygomycetes. Examples of bacteria that can cause neuroinflammation include Listeria monocytogenes, Borrelia burgdorferi, Neisseria meningitidis, Streptococcus pneumoniae, Staphylococcus aureus, Haemophilus influenzae, Mycobacterium, and Brucella species. Neuroinflammation can also be associated with parasitic infections.

[0051] In certain embodiments, neuroinflammation results from systemic inflammation from the subject's peripheral immune system caused by an infectious disease such as COVID or "long COVID." In this embodiment, treatment of a subject with the herbal compositions described herein, including treatment with MLC901 or treatment with MLC1501, provides protection against the progression of the neurodegenerative process by regulating, modulating, protecting, and / or strengthening the blood-brain barrier against the damaging effects of neuroinflammation.

[0052] In one embodiment of any of the above aspects, the neuroinflammation is the result of exposure to a toxin. In one embodiment of any of the above aspects, neuroinflammation is associated with neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, Huntington's disease, frontotemporal dementia, amyotrophic lateral sclerosis, multiple sclerosis, 10-12 primary tauopathies, synucleinopathies (i.e., dementia with Lewy bodies and multiple system atrophy), polyglutamine diseases including spinocerebellar ataxias, prion diseases, traumatic brain injury, chronic traumatic encephalopathy, stroke, and spinal cord injury.

[0053] In one embodiment, neuroinflammation is associated with a neuropsychiatric condition such as depression, stress, post-traumatic stress disorder, schizophrenia, autism spectrum disorder, or epilepsy. In one embodiment of any of the above aspects, the neuroinflammation is associated with aging (e.g., "inflammagenesis").

[0054] Other disorders that can cause neuroinflammation include postoperative cognitive dysfunction, ischemia, brain tumors, chronic pain, headache, migraine, trigeminal neuralgia, cardiovascular disease and associated risk factors (e.g., due to obesity, hypertension, diabetes, hyperlipidemia, smoking, etc.), chronic peripheral inflammatory conditions (e.g., rheumatoid arthritis, inflammatory bowel disease, psoriasis, etc.), perturbation of the gut microbiota (i.e., dysbiosis), and herpes keratitis.

[0055] In one embodiment, neuroinflammation arises due to systemic inflammation from the subject's peripheral immune system caused by perturbation of the gut microbiota (i.e., dysbiosis). In this embodiment, treatment of a subject with the herbal compositions described herein, including treatment with MLC901 or treatment with MLC1501, provides protection against the progression of the neurodegenerative process by regulating, modulating, protecting, and / or strengthening the blood-brain barrier against the damaging effects of neuroinflammation.

[0056] In one embodiment of any of the above aspects, the neuroinflammation results from chemical intervention to treat the underlying disease or disorder. One example of such chemical intervention is the use of TNF inhibitors to treat autoimmune diseases such as rheumatoid arthritis.

[0057] In one embodiment of any of the above aspects, the neuroinflammation results from a mechanical intervention to treat an underlying disease or disorder. In one embodiment of any of the above aspects, the neuroinflammation results from surgical intervention to treat an underlying condition or disease. Examples of surgical intervention include carotid endarterectomy, carotid angioplasty with or without stenting, vertebral angioplasty with or without stenting, intracranial angioplasty with or without stenting, aneurysm coiling, occlusion of vascular malformations, repair of intracranial fistulas, thrombectomy, extracranial-intracranial bypass, open-heart surgery, cardiac surgery requiring bypass surgery, coronary artery bypass grafting, coronary angioplasty with or without stenting, valve surgery / valvuloplasty, valvotomy, closure of patent foramen ovale, and surgery on the great vessels of the heart.

[0058] In one embodiment of any of the above aspects, the effective amount of the herbal composition for treating neuroinflammation is from about 1 mg / kg to about 100 mg / kg. In one embodiment of any of the above aspects, there is provided a method comprising administering the herbal composition herein for treating neuroinflammation and a second agent for treating the underlying disease or disorder.

[0059] In one embodiment of any of the above aspects, the subject is diagnosed with neuroinflammation, such as by a healthcare professional. In one embodiment of any of the above aspects, the subject is tested for neuroinflammation by measuring the levels of pro-inflammatory chemokines and / or cytokines, for example, the levels of one or more cytokines selected from IL-1β, IL-6, and TNF-α.

[0060] In one embodiment of any of the above embodiments, the subject is tested for neuroinflammation by measuring levels of inflammasomes comprising NLRP3 and / or iNOS. In one aspect, a subject who is confirmed after testing to have neuroinflammation (e.g., by identifying the subject as exhibiting elevated levels of pro-inflammatory chemokines and / or cytokines, e.g., elevated levels of one or more cytokines selected from IL-1β, IL-6, and TNF-α, and / or elevated levels of one or more inflammasomes, e.g., NLRP3 and / or iNOS) is treated with an effective amount of an herbal composition comprising at least four herbal ingredients, Polygalae Root (Radix Polygalae), Astragali Root (Radix Astragali), Rhizome Ligusticum Chuanxiong, and Angelica Root (Radix Angelica sinensis), or extracts thereof.

[0061] In a further aspect, subjects who are confirmed to have neuroinflammation after testing (e.g., by identifying the subject as exhibiting elevated levels of pro-inflammatory chemokines and / or cytokines, e.g., elevated levels of one or more cytokines selected from IL-1β, IL-6, and TNF-α, and / or elevated levels of one or more inflammasomes, e.g., NLRP3 and / or iNOS) are treated with a combination of four herbal ingredients, Polygala Root (Radix Polygalae) (Root of Japanese Bush Clover), Radix Astragali (Root of Astragali), Cnidium Rhizome (Rhizome Ligusticum Chuanxiong), and Radix Angelica Sinensis (Root of Chinese Angelica) or extracts thereof, and Radix Salviae Miltiorrhizae (Red Sage Root), Radix Paeoniae Radix (Sesame Root), and Radix Paeoniae Radix (Sesame Root), or extracts thereof. and at least one additional herbal ingredient selected from the group consisting of Red Peony Root (Rubra), Carthamus Tinctorius (Safflower) Flower, Semen Persicae (Peach (Prunus Persica) Seed), and Rhizome Acori Tatarinowii (Rhizome of the Japanese Calamus) or extracts thereof.

[0062] In another aspect, a subject who is confirmed after testing to have neuroinflammation (e.g., by identifying the subject as exhibiting elevated levels of pro-inflammatory chemokines and / or cytokines, e.g., elevated levels of one or more cytokines selected from IL-1β, IL-6, and TNF-α, and / or elevated levels of one or more inflammasomes, e.g., NLRP3 and / or iNOS) is treated with an effective amount of an herbal composition consisting essentially of at least four herbal ingredients, Polygala Root (Radix Polygalae) (Root of Japanese Bush Clover), Radix Astragali (Root of Astragali), Rhizome Ligusticum Chuanxiong, and Radix Angelica sinensis (Root of Angelica Root), or extracts thereof.

[0063] In yet another aspect, subjects who are confirmed to have neuroinflammation after testing (e.g., by identifying the subject as exhibiting elevated levels of pro-inflammatory chemokines and / or cytokines, e.g., elevated levels of one or more cytokines selected from IL-1β, IL-6, and TNF-α, and / or elevated levels of one or more inflammasomes, e.g., NLRP3 and / or iNOS) are those who have been identified as having ...) and / or elevated The patient is treated with an effective amount of an herbal composition consisting essentially of (a) red peony root (Rubra), (b) safflower (Carthamus Tinctorius) flowers (Safflower), (c) peach seeds (Semen Persicae) (Peach (Prunus Persica) seeds), and (d) iris rhizome (Rhizome Acori Tatarinowii) (Rhizome of the Stone Calamus) or extracts thereof.

[0064] In one embodiment of any of the above aspects, the herbal composition comprises an extract of the herbal ingredients described above. In one embodiment of any of the above aspects, the herbal composition comprises 5 times the weight to weight ratio of Radix Astragali or an extract thereof compared to the weight of each of the other herbal ingredients or extracts thereof present.

[0065] In one embodiment of any of the above aspects where the subject is confirmed to have neuroinflammation after testing, the herbal composition used to treat the subject is MLC901. In one embodiment of any of the above aspects, where the subject is confirmed to have neuroinflammation after testing, the herbal composition used to treat the subject is MLC1501.

[0066] In one aspect, a subject at risk for neuroinflammation as a result of injury (e.g., due to the aforementioned conditions, disease, injury, infection, age, or chemical, mechanical, or surgical intervention) is treated to prevent or limit neuroinflammation by administering (before, at the time of, or after the injury) an effective amount of an herbal composition comprising at least four herbal ingredients: Radix Polygalae (Primula Root), Radix Astragali (Astragalus Root), Rhizome Ligusticum Chuanxiong (Cnidium Rhizome), and Radix Angelica sinensis (Angelica Root), or extracts thereof.

[0067] In a further aspect, subjects at risk for neuroinflammation as a result of injury (e.g., due to the aforementioned conditions, disease, injury, infection, age, or chemical, mechanical, or surgical intervention) are treated with four herbal ingredients, Polygala Root (Radix Polygalae) (Root of the Japanese bush clover), Astragalus Root (Radix Astragali) (Root of Astragali), Cnidium Rhizome (Rhizome Ligusticum Chuanxiong), and Angelica Root (Radix Angelica sinensis) (Root of the Chinese angelica tree) or extracts thereof, in combination with Danshen Root and Rhizome (Radix et Rhizome Salviae Miltiorrhizae) (Red sage root), Radix Paeoniae Rubra (Red peony root), Carthamus Tinctorius (Safflower) flower, and Peach Seed (Semen Persicae) (Peach) The subject is treated to prevent or limit neuroinflammation by administering (before, at the time of, or after the injury) an effective amount of an herbal composition comprising: (a) a herbal composition selected from the group consisting of: (b) a herbal composition containing: (c) a herbal composition containing: (d) a herbal composition containing: (e) a herbal composition containing: (f) a herbal composition containing: (f) a herbal composition containing: (g ...

[0068] In another aspect, a subject at risk for neuroinflammation as a result of injury (e.g., due to the aforementioned conditions, disease, injury, infection, age, or chemical, mechanical, or surgical intervention) is treated to prevent or limit neuroinflammation by administering (before, at the time of, or after the injury) an effective amount of an herbal composition consisting essentially of at least four herbal ingredients: Radix Polygalae (Primula Root), Radix Astragali (Astragalus Root), Rhizome Ligusticum Chuanxiong (Cnidium Rhizome), and Radix Angelica sinensis (Angelica Root), or extracts thereof.

[0069] In yet another aspect, a subject at risk for neuroinflammation as a result of injury (e.g., due to the aforementioned conditions, disease, injury, infection, age, or chemical, mechanical, or surgical intervention) is receiving a neuroinflammation treatment containing at least nine herbal ingredients: Polygalae Root (Radix Polygalae) (Root of the Japanese bush clover), Astragalus Root (Radix Astragali) (Root of Astragali), Cnidium Rhizome (Rhizome Ligusticum Chuanxiong), Angelica Root (Radix Angelica sinensis) (Root of the Chinese angelica tree), Salvia Root and Rhizome (Radix et Rhizome Salviae Miltiorrhizae) (Red sage root), Radix Paeoniae Rubra (Red peony root), Carthamus Tinctorius (Safflower) flower, Peach Seed (Semen Persicae) (Peach (Prunus Persica) seed), and Rhizome Acori The present invention relates to a method for treating neuroinflammation in a subject, the method comprising administering (before, at the time of, or after injury) an effective amount of an herbal composition consisting essentially of A. tatarinowii (the rhizome of the calamus plant) or an extract thereof to the subject to prevent or limit neuroinflammation.

[0070] In one embodiment of any of the above aspects, the herbal composition comprises an extract of the herbal ingredients described above. In one embodiment of any of the above aspects, the herbal composition comprises 5 times the weight to weight ratio of Radix Astragali or an extract thereof compared to the weight of each of the other herbal ingredients or extracts thereof present.

[0071] In one embodiment of any of the above aspects where the subject is at risk of neuroinflammation as a result of injury, the herbal composition used to prevent or limit neuroinflammation is MLC901. In one embodiment of any of the above aspects where the subject is at risk of neuroinflammation as a result of injury, the herbal composition used to prevent or limit neuroinflammation is MLC1501.

[0072] In one aspect, a subject at risk of developing a disease or condition as a result of neuroinflammation is treated with an effective anti-inflammatory amount of an herbal composition comprising at least four herbal ingredients, Radix Polygalae (Primula Root), Radix Astragali (Astragalus Root), Rhizome Ligusticum Chuanxiong, and Radix Angelica sinensis (Angelica Root), or extracts thereof, to prevent or alleviate the onset of the disease or condition.

[0073] In a further aspect, a subject at risk of developing a disease or condition as a result of neuroinflammation may be administered a combination of four herbal ingredients, Polygala Root (Radix Polygalae) (Root of the Japanese bush clover), Astragalus Root (Radix Astragali) (Root of Astragali), Cnidium Rhizome (Rhizome Ligusticum Chuanxiong), and Angelica Root (Radix Angelica sinensis) (Root of the Chinese angelica tree) or extracts thereof, with Salvia Root and Rhizome (Radix et Rhizome Salviae Miltiorrhizae) (Red sage root), Paeoniae Root (Radix Paeoniae Rubra) (Red peony root), Carthamus Tinctorius (Safflower) flower, Peach Seed (Semen Persicae) (Peach (Prunus Persica) seed), and Rhizome Acori (Rhizome Acori) ... and at least one additional herbal ingredient selected from the group consisting of: Calamus tatarinowii (rhizome of the calamus plant) or extracts thereof.

[0074] In another aspect, a subject at risk of developing a disease or condition as a result of neuroinflammation is treated with an effective anti-inflammatory amount of an herbal composition consisting essentially of at least four herbal ingredients, Radix Polygalae (Primula Root), Radix Astragali (Astragalus Root), Rhizome Ligusticum Chuanxiong, and Radix Angelica sinensis (Angelica Root), or extracts thereof, to prevent or alleviate the onset of the disease or condition.

[0075] In yet another aspect, a subject at risk of developing a disease or condition as a result of neuroinflammation is administered a combination of at least nine herbal ingredients, Polygalae Root (Radix Polygalae) (Root of the Japanese bush clover), Astragali Root (Radix Astragali) (Root of Astragali), Cnidium Rhizome (Rhizome Ligusticum Chuanxiong), Angelica Root (Radix Angelica sinensis) (Root of the Chinese angelica tree), Danshen Root and Rhizome (Radix et Rhizome Salviae Miltiorrhizae) (Red sage root), Radix Paeoniae Rubra (Red peony root), Carthamus Tinctorius (Safflower) flower, Semen Persicae (Peach seed), and Rhizome Acori (Rhizome Acori) to prevent or alleviate the onset of the disease or condition. The patient is treated with an effective anti-inflammatory amount of a herbal composition consisting essentially of Araceae (Rhizome of Calamus tatarinowii) or an extract thereof.

[0076] In one embodiment of any of the above aspects, the herbal composition comprises an extract of the herbal ingredients described above. In one embodiment of any of the above aspects, the herbal composition comprises 5 times the weight to weight ratio of Radix Astragali or an extract thereof compared to the weight of each of the other herbal ingredients or extracts thereof present.

[0077] In one embodiment of any of the above aspects where the subject is at risk of developing a disease or condition as a result of neuroinflammation, the herbal composition used to prevent or alleviate the onset of the disease or condition is MLC901.

[0078] In one embodiment of any of the above aspects where the subject is at risk of developing a disease or condition as a result of neuroinflammation, the herbal composition used to prevent or alleviate the onset of the disease or condition is MLC1501.

[0079] The "disease or condition" may be, for example, brain injury, brain swelling, leakage of the blood-brain barrier, stroke, transient ischemic attack, or a neuropsychiatric disorder (e.g., depression, stress, post-traumatic stress disorder, schizophrenia, autism spectrum disorder, or epilepsy).

[0080] NeuroAid II, referred to herein as "MLC901," is a composition containing extracts of nine herbal ingredients (Astragalus membranaceus root (Radix Astragali), Danshen (Radix Salvia miltiorrhizae), Paeonia rubra root (Radix Paeoniae rubra), Cnidium rhizome (Rhizoma Ligusticum Chuanxiong), Angelica sinensis root (Radix Angelica sinensis), Safflower (Carthamus Tinctorius), Peach (Prunus Persica), Polygala root (Radix Polygalae), and Paeonia suffruticosa rhizome (Rhizoma Acori Tatarinowii)). In MLC901, Astragalus membranaceus root (Radix Astragali) extract is present at five times the weight-to-weight ratio of each of the other herbal ingredient extracts present. It is currently marketed as an oral treatment to support recovery after stroke.

[0081] MLC1501 is a composition containing extracts of four herbal ingredients (Astragali root (Radix Astragali), Cnidium rhizome (Rhizoma Ligusticum Chuanxiong), Polygala root (Radix Polygalae), and Angelica sinensis root (Radix Angelica sinensis). In MLC1501, Astragali root (Radix Astragali) extract is present at five times the weight-to-weight ratio of each of the other herbal ingredient extracts present. It is currently undergoing human clinical trials to evaluate its effectiveness in post-stroke recovery.

[0082] MLC901 and MLC1501, and their preparation, are described in published PCT application WO 2017 / 048191 A1, the contents of which are incorporated herein by reference in their entirety.

[0083] In one embodiment of any of the above aspects, the herbal composition of the methods of the present invention consists essentially of four herbal ingredients, Polygalae root (Radix Polygalae), Astragalus membranaceus root (Radix Astragali), Cnidium rhizome (Rhizome Ligusticum Chuanxiong), and Angelica quince root (Radix Angelica sinensis), or extracts thereof, in combination with one, two, three, four, or five herbal ingredients, Danshen root (Radix Salvia Miltiorrhizae), Watercress root (Radix Paeoniae Rubra), Safflower (Carthamus Tinctorius), Peach (Prunus Persica), and Red Pepper rhizome (Rhizoma Acori tatarinowii), or extracts thereof. Such herbal compositions and their preparation are described in published PCT applications WO 2007 / 106049 A1, WO 2010 / 053456 A1, WO 2010 / 110755 A1, and WO 2013 / 141818 A1, the contents of which are incorporated by reference herein in their entireties.

[0084] The herbal compositions herein containing MLC901 and MLC1501 are generally highly effective in treating or alleviating neuroinflammation in subjects who have had a stroke, particularly an ischemic stroke, and aid in post-stroke recovery. However, patients with underlying conditions, particularly those with an inflammatory component such as diabetes mellitus, are known to suffer from poor prognosis for functional recovery after stroke and standard treatment, including rehabilitation therapy. Stroke in diabetic subjects results in increased mortality and long-term neurological and functional impairment with standard treatment. Nevertheless, the herbal compositions herein containing MLC901 and MLC1501 have been found to effectively aid post-stroke recovery in diabetic subjects.

[0085] Thus, in one aspect of the present disclosure, there is provided a method of treating or alleviating neuroinflammation in a diabetic subject following an ischemic stroke, comprising administering to the subject an effective amount of an herbal composition comprising at least four herbal ingredients: Polygalae (Primarinus officinalis), Astragali (Astragalus membranaceus), Ligusticum Chuanxiong (Cnidium rhizome), and Angelica sinensis (Angelica sinensis), their roots or rhizomes, or extracts thereof.

[0086] In one embodiment of the above aspect, the herbal composition comprising four herbal ingredients, Polygalae (Princess bush clover), Astragali (Astragali), Ligusticum Chuanxiong, and Angelica sinensis (Angelica sinensis), their roots or rhizomes, or extracts thereof, further comprises at least one herbal composition selected from the group consisting of Salviae Miltiorrhizae (Red sage), Paeoniae Rubra (Red peony), Carthamus Tinctorius (Safflower) flower, Semen Persicae (Peach (Prunus Persica) seed), and Acori Tatarinowii (Stone iris), their roots or rhizomes, or extracts thereof.

[0087] In a further aspect, there is provided a method of treating or alleviating neuroinflammation in a diabetic subject following an ischemic stroke, comprising administering to the subject an effective amount of an herbal composition consisting essentially of four herbal ingredients: Polygalae (Primarinus officinalis), Astragali (Astragalus membranaceus), Ligusticum Chuanxiong (Cnidium Root), and Angelica sinensis (Angelica sinensis), their roots or rhizomes, or extracts thereof.

[0088] In another aspect, there is provided a method of treating or alleviating neuroinflammation in a diabetic subject following an ischemic stroke, comprising administering to the subject an effective amount of an herbal composition consisting essentially of nine herbal ingredients Polygalae (Plumeriana tenuifolia), Astragali (Astragali root), Ligusticum Chuanxiong (Cnidium rhizome), Angelica sinensis (Angelica sinensis), Salviae Miltiorrhizae (Red sage), Paeoniae Rubra (Red peony), Carthamus Tinctorius (Safflower) flower, Semen Persicae (Peach seed), and Acori Tatarinowii (Stone iris), their roots or rhizomes, or extracts thereof.

[0089] In one embodiment of any of the above aspects, the herbal composition is a pharmaceutical composition. The pharmaceutical composition may also include one or more pharmaceutically acceptable carriers or excipients. In a specific embodiment, the herbal composition is MLC901. In a specific embodiment, the herbal composition is MLC1501.

[0090] In certain aspects of the present disclosure, there is provided a method of treating or alleviating neuroinflammation in a diabetic subject following an ischemic stroke, comprising administering to the subject an effective amount of MLC901. Certain aspects of the present disclosure provide methods for treating or alleviating neuroinflammation in a diabetic subject following an ischemic stroke, comprising administering to the subject an effective amount of MLC1501.

[0091] Certain aspects of the present disclosure provide methods for treating a diabetic subject after an ischemic stroke, comprising administering to the subject an effective amount of MLC901 to improve post-stroke recovery. In one embodiment, the subject is receiving post-stroke standard treatment, such as treatment with an antiplatelet agent (e.g., aspirin) and / or rehabilitation therapy. In one embodiment, diabetic subjects receiving post-stroke standard treatment with co-administered MLC901 exhibit superior recovery rates compared to diabetic subjects receiving post-stroke standard treatment alone. In one embodiment, the superiority is demonstrated within about 3, 6, 9, 12, 18, or 24 months after the stroke occurred.

[0092] Certain aspects of the present disclosure provide methods for treating a diabetic subject after an ischemic stroke, comprising administering to the subject an effective amount of MLC1501 to improve post-stroke recovery. In one embodiment, the subject is receiving post-stroke standard treatment, such as treatment with an antiplatelet agent (e.g., aspirin) and / or rehabilitation therapy. In one embodiment, diabetic subjects receiving post-stroke standard treatment with co-administered MLC1501 exhibit superior recovery rates compared to diabetic subjects receiving post-stroke standard treatment alone. In one embodiment, the superiority is demonstrated within about 3, 6, 9, 12, 18, or 24 months after the stroke occurred.

[0093] The pharmaceutical compositions of the present invention may optionally comprise one or more pharmaceutically acceptable additives, carriers, and / or diluents. Examples of pharmaceutically acceptable additives include pharmaceutically acceptable excipients, buffers, adjuvants, stabilizers, diluents, fillers, preservatives, lubricants, or other pharmaceutically acceptable substances known to those of skill in the art or described herein. Examples of suitable pharmaceutical carriers or diluents include phosphate-buffered saline, water, emulsions (such as oil / water emulsions), various types of wetting agents, sterile solutions, etc. Examples of excipients that can be used include, for example, sugars, starches, celluloses, gums, proteins, dextrins, and maltodextrins. Various formulations are generally known and are described in detail in the latest edition of Remington's Pharmaceutical Sciences (Maack Publishing, Easton PA). In at least some embodiments, the pharmaceutical compositions described herein include an excipient such as, for example, dextrin or maltodextrin.

[0094] In one embodiment of any of the above aspects, the compositions (e.g., pharmaceutical compositions) of the present disclosure may be included in a kit. The kit may also include instructions for use in addition to the herbal ingredients. The kit may be advertised, distributed, and / or sold as a unit for performing one of the aspects of the present disclosure.

[0095] In general, the pharmaceutical compositions of the present disclosure may be prepared according to methods known to those skilled in the art. The composition may be, for example, a solution, a suspension, liquid, chopped herbs, powder, a paste, aqueous, non-aqueous, or any combination thereof.

[0096] Compositions (e.g., pharmaceutical compositions) of the present disclosure may be administered by any suitable route, such as orally, parenterally, intravenously, subcutaneously, intradermally, intraperitoneally, or topically, in liquid, semi-liquid, or solid form, and are formulated in a manner appropriate for each administration route. The term "administering" and variations thereof, including "administer" and "administration," include contacting, applying, delivering, or providing a composition of the present disclosure to an organism or surface by any suitable means.

[0097] Herbal ingredients may be administered in therapeutically effective amounts (single dose or as part of multiple doses). "Effective amount" or "therapeutically effective amount" means the amount administered to achieve physiological significance. An agent is physiologically significant when present in an amount that produces a detectable change in the physiology of the recipient patient such that a beneficial or desired result is achieved.

[0098] The exact amount required will vary from subject to subject, depending on factors such as the species being treated, the subject's age, weight, and general health, the condition being treated and its severity, the mode of administration, the subject's sex, diet, the time and frequency of administration, drug combinations, and tolerance / response to treatment.

[0099] In one embodiment of any of the above aspects, the composition (e.g., pharmaceutical composition) of the present disclosure is administered as a capsule taken one or more times daily (e.g., 1, 2, 3, or 4 times). In a particular embodiment, MLC901 is administered in the form of two capsules taken three times daily. In a particular embodiment, MLC1501 is administered in the form of four capsules taken twice daily. For patients with swallowing difficulties, the capsules may be opened and the powder diluted with water and can be swallowed directly or injected through a stomach tube.

[0100] In one embodiment of any of the above aspects, the daily dose for a patient is from about 500 mg to about 8 g, for example, from about 1 g to about 8 g (including about 1 g, 2 g, 3 g, 4 g, 5 g, 6 g, 7 g, or 8 g). A "daily dose" can be a single unit dose (e.g., tablet, capsule, etc.) taken on a particular day, or multiple unit doses. However, it should be understood that dosages may vary depending on, for example, the requirements of the patient and the severity of the condition being treated.

[0101] The duration of treatment is adaptive based on the patient's condition, be it a few days or more typically several months (e.g., 3 months). In one embodiment of any of the above aspects, treatment lasts for about 12 weeks. In another embodiment of any of the above aspects, treatment lasts for about 24 weeks. In another embodiment of any of the above aspects, treatment lasts for about 36 weeks. In another embodiment of any of the above aspects, treatment lasts for about 48 weeks. In another embodiment of any of the above aspects, treatment lasts for more than about 48 weeks.

[0102] The timing of administration of the first dose of the herbal composition of the present disclosure (e.g., MLC901 or MLC1501) should generally occur as soon as possible after an injury, e.g., caused by injury, infection, exposure to a toxin, neurodegenerative disease, aging (e.g., "inflammage-aging"), or due to chemical, mechanical, or surgical intervention to treat an underlying disease or disorder that may cause neuroinflammation. For example, the first dose can be administered within 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 days (e.g., within 1-5 days, 1-4 days, 1-3 days, or 1-2 days) after the injury. In one embodiment, the first dose of the herbal composition of the present disclosure (e.g., MLC901 or MLC1501) is administered within 24 hours after the injury, e.g., within 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 hours after the injury.

[0103] In one embodiment of any of the above aspects, the treatment regimen is designed for acute treatment. In one embodiment of any of the above aspects, the treatment regimen is designed for chronic treatment. In one embodiment of any of the above aspects, the treatment regimen is designed for post-acute treatment.

[0104] In one embodiment of any of the above aspects, the herbal composition of the present disclosure (e.g., MLC901 or MLC1501) may be administered to a subject in combination with an active agent for treating an underlying disease or disorder and / or in combination with an additional anti-inflammatory therapy (e.g., one or more anti-inflammatory agents). The one or more additional active agents may be administered at the same time (e.g., simultaneously) as the herbal composition of the present disclosure, or at different times (e.g., sequentially), and over different time periods that may be separate from or overlapping with each other.

[0105] The one or more additional active agents may be administered via the same route as the herbal composition of the present disclosure or via a different route. The one or more additional active agents and the appropriate route of administration and dosage level are known to, or can be easily determined by, those skilled in the art. Typically, as is well known in the medical field, the administration regimen may depend on various factors, including the patient's size, body surface area, age, the specific compound being administered, sex, time and route of administration, general health, and other concurrently administered medications. While individual needs vary, determining the optimal range of effective amounts of each component is within the skill of one of ordinary skill in the art. Typically, the dosage of the one or more additional active agents is the same or similar to the amount that would be administered if the agent were used without the herbal composition of the present disclosure. When the herbal composition of the present disclosure is administered with one or more additional active agents, the one or more additional active agents may be provided in a composition or kit that includes the herbal composition of the present disclosure, or the one or more additional active agents may be provided separately (i.e., not part of a composition or kit that provides the herbal composition of the present disclosure).

[0106] In addition to the embodiments described above, the present disclosure includes the following specific embodiments. Embodiment 1: A herbal composition comprising at least four herbal ingredients, Polygala Root (Radix Polygalae) (Root of Lady's Bush), Astragalus Root (Radix Astragali), Cnidium Rhizome (Rizome Ligusticum Chuanxiong), and Angelica Root (Radix Angelica sinensis) (Root of Angelica sinensis) or extracts thereof for use in treating neuroinflammation in a subject. Embodiment 2: The herbal composition for use according to embodiment 1, wherein the herbal composition further comprises at least one herbal ingredient selected from the group of Radix et Rhizome Salviae Miltiorrhizae (red sage root), Radix Paeoniae Rubra (red peony root), Carthamus Tinctorius (safflower), Semen Persicae (peach seed), and Rhizome Acori Tatarinowii (iris rhizome), or an extract thereof. Embodiment 3: The herbal composition for use according to embodiment 1, wherein the herbal composition consists essentially of at least four herbal ingredients, Polygala Root (Radix Polygalae) (Root of Lady's Bush), Astragalus Root (Radix Astragali), Cnidium Rhizome (Rizome Ligusticum Chuanxiong), and Angelica Root (Radix Angelica sinensis) (Root of Angelica sinensis) or extracts thereof. Embodiment 4: The herbal composition for use according to embodiment 1, wherein the herbal composition consists essentially of at least nine herbal ingredients, Polygala Root (Radix Polygalae) (Root of Japanese bush clover), Astragalus Root (Radix Astragali) (Root of Astragali), Cnidium Rhizome (Rizome Ligusticum Chuanxiong), and Angelica Root (Radix Angelica sinensis) (Root of Chinese angelica), Salvia Root and Rhizome (Radix et Rhizome Salviae Miltiorrhizae) (Red sage root), Radix Paeoniae Rubra (Red peony root), Carthamus Tinctorius (Safflower) flower, Peach Seed (Semen Persicae) (Peach (Prunus Persica) seed), and Rhizome Acori Tatarinowii (Rhizome of Japanese iris) or extracts thereof.

[0107] Embodiment 5: The herbal composition for use according to any one of embodiments 1 to 4, wherein the herbal composition is a pharmaceutical composition. Embodiment 6: The herbal composition for use according to embodiment 5, wherein the pharmaceutical composition also comprises a pharmaceutically acceptable carrier or excipient.

[0108] Embodiment 7: The herbal composition for use according to embodiment 6, wherein the pharmaceutical composition is composition MLC901. Embodiment 8: The herbal composition for use according to embodiment 6, wherein the pharmaceutical composition is composition MLC1501.

[0109] Embodiment 9: The herbal composition for use according to any one of embodiments 1 to 8, wherein the effective amount of the herbal composition is from about 1 mg / kg to about 100 mg / kg. Embodiment 10: The herbal composition for use according to any one of embodiments 1 to 9, wherein the method comprises administering a second anti-inflammatory agent.

[0110] Embodiment 11: The herbal composition for use according to any one of embodiments 1 to 10, wherein the neuroinflammation is the result of an injury. Embodiment 12: The herbal composition for use according to embodiment 11, wherein the injury is selected from the group consisting of spinal cord injury, stroke, traumatic brain injury, subarachnoid hemorrhage, subdural hemorrhage or epidural hemorrhage.

[0111] Embodiment 13: The herbal composition for use according to any one of embodiments 1 to 10, wherein the neuroinflammation is a result of an infection. Embodiment 14: The herbal composition for use according to embodiment 13, wherein the infection is an infection of the brain or central nervous system (e.g., encephalitis or bacterial, viral or fungal meningitis).

[0112] Embodiment 15: The infection is a viral infection (e.g., a varicella-zoster virus (VZV) infection, a herpes simplex virus (HSV-1 or HSV-2), SARS-CoV-2 virus or a variant thereof, Epstein-Barr virus, hepatitis B virus, hepatitis C virus, human immunodeficiency virus type 1 (HIV-1), human papillomavirus (HPV), human T-cell lymphotropic virus type 1 (HTLV-1), or Kaposi's sarcoma herpesvirus (KSHV)), a fungal infection (e.g., a fungus such as Cryptococcus neoformans, Candida albicans, Histoplasma capsulatum, Coccidioides immitis, Paracoccidioides spp., or ... brasiliensis, Aspergillus spp. and zygomycetes], bacterial infection (for example, the bacteria is selected from the group consisting of Listeria monocytogenes, Borrelia burgdorferi, Neisseria meningitidis, Streptococcus pneumoniae, Staphylococcus aureus, Haemophilus influenzae, Mycobacterium and Brucella spp.) or parasitic infection.

[0113] Embodiment 16: The herbal composition for use according to any one of embodiments 1 to 10, wherein the neuroinflammation is a result of exposure to a toxin. Embodiment 17: The herbal composition for use according to any one of embodiments 1 to 10, wherein the neuroinflammation is associated with neurodegenerative diseases (e.g., Alzheimer's disease, Parkinson's disease, Huntington's disease, frontotemporal dementia, amyotrophic lateral sclerosis, multiple sclerosis, 10-12 primary tauopathies, synucleinopathies (i.e., dementia with Lewy bodies and multiple system atrophy), polyglutamine diseases including spinocerebellar ataxias, prion diseases, traumatic brain injury, chronic traumatic encephalopathy, stroke and spinal cord injury).

[0114] Embodiment 18: The herbal composition for use according to any one of embodiments 1 to 10, wherein the neuroinflammation is associated with a neuropsychiatric condition (e.g., depression, stress, post-traumatic stress disorder, schizophrenia, autism spectrum disorder or epilepsy).

[0115] Embodiment 19: The herbal composition for use according to any one of embodiments 1 to 10, wherein neuroinflammation is associated with aging (e.g., "inflammagenesis"). Embodiment 20: The herbal composition for use according to any one of embodiments 1 to 10, wherein the neuroinflammation is associated with a disease or disorder selected from the group consisting of post-operative cognitive dysfunction, ischemia, brain tumor, chronic pain, headache, migraine, trigeminal neuralgia, cardiovascular disease and associated risk factors (e.g., due to obesity, hypertension, diabetes, hyperlipidemia, smoking, etc.), chronic peripheral inflammatory conditions (e.g., rheumatoid arthritis, inflammatory bowel disease, psoriasis, etc.), perturbation of the gut microbiota (i.e., dysbiosis), and herpes keratitis.

[0116] Embodiment 21: The herbal composition for use according to any one of embodiments 1 to 10, wherein the neuroinflammation results from a chemical intervention to treat an underlying disease or disorder. An example of such a chemical intervention is the use of TNF inhibitors to treat autoimmune diseases such as rheumatoid arthritis.

[0117] Embodiment 22: The herbal composition for use according to any one of embodiments 1 to 10, wherein the neuroinflammation results from an underlying disease or mechanical intervention to treat the disease. Embodiment 23: The herbal composition for use according to any one of embodiments 1 to 10, wherein the neuroinflammation results from an underlying disease or surgical intervention to treat the disease.

[0118] Embodiment 24: The herbal composition for use according to embodiment 23, wherein the surgical intervention is selected from the group consisting of carotid endarterectomy, carotid angioplasty with or without stenting, vertebral angioplasty with or without stenting, intracranial angioplasty with or without stenting, aneurysm coiling, occlusion of vascular malformations, repair of intracranial fistulas, thrombectomy, extracranial-intracranial bypass, open heart surgery, cardiac surgery requiring bypass surgery, coronary artery bypass grafting, coronary angioplasty with or without stenting, valve surgery / valvuloplasty, valvulotomy, closure of patent foramen ovale, and surgery on the great vessels of the heart.

[0119] Embodiment 25: The herbal composition for use according to any one of embodiments 21 to 24, wherein the herbal composition is administered before, simultaneously with, or after the administration of an intervention to treat the underlying disease or disorder.

[0120] Embodiment 26: The herbal composition for use according to any one of embodiments 1 to 25, wherein the subject is tested for and confirmed to have neuroinflammation prior to administering the herbal composition.

[0121] Embodiment 27: The herbal composition for use according to embodiment 26, wherein testing shows that the subject presents high levels of pro-inflammatory chemokines and / or cytokines. Embodiment 28: The herbal composition for use according to embodiment 26, wherein testing shows that the subject presents elevated levels of one or more pro-inflammatory chemokines selected from IL-1β, IL-6 and TNF-α and / or elevated levels of one or more inflammasomes selected from NLRP3 and iNOS.

[0122] Embodiment 29: A herbal composition comprising at least four herbal ingredients, Radix Polygalae (Primula Root), Radix Astragali (Astragalus Root), Rhizome Ligusticum Chuanxiong, and Radix Angelica sinensis (Angelica Root), or extracts thereof, for use in preventing or limiting neuroinflammation in a subject at risk of neuroinflammation as a result of injury. Embodiment 30: The herbal composition for use according to embodiment 29, wherein the herbal composition further comprises at least one herbal ingredient selected from the group of Radix et Rhizome Salviae Miltiorrhizae (red sage root), Radix Paeoniae Rubra (red peony root), Carthamus Tinctorius (safflower), Semen Persicae (peach seed), and Rhizome Acori Tatarinowii (iris rhizome), or an extract thereof. Embodiment 31: A herbal composition for use according to embodiment 29, consisting essentially of at least four herbal ingredients, Polygala Root (Radix Polygalae) (Root of Lady's Bush), Astragalus Root (Radix Astragali), Cnidium Rhizome (Rizome Ligusticum Chuanxiong), and Angelica Root (Radix Angelica sinensis) (Root of Angelica Root), or extracts thereof. Embodiment 32: The herbal composition for use according to embodiment 29, wherein the herbal composition consists essentially of at least nine herbal ingredients, Polygala Root (Radix Polygalae) (Root of Japanese Bush Clover), Astragalus Root (Radix Astragali) (Root of Astragali), Cnidium Rhizome (Rizome Ligusticum Chuanxiong) and Angelica Root (Radix Angelica sinensis) (Root of Chinese Angelica), Salvia Root and Rhizome (Radix et Rhizome Salviae Miltiorrhizae) (Red Sage Root), Radix Paeoniae Rubra (Red Peony Root), Carthamus Tinctorius (Safflower) Flower, Peach Seed (Semen Persicae) (Peach (Prunus Persica) Seed), and Rhizome Acori Tatarinowii (Rhizome of Stone Calamus) or extracts thereof.

[0123] Embodiment 33: The herbal composition for use according to any one of embodiments 29 to 32, wherein the herbal composition is a pharmaceutical composition. Embodiment 34: The herbal composition for use according to embodiment 33, wherein the pharmaceutical composition also comprises a pharmaceutically acceptable carrier or excipient.

[0124] Embodiment 35: The herbal composition for use according to embodiment 34, wherein the pharmaceutical composition is composition MLC901. Embodiment 36: The herbal composition for use according to embodiment 34, wherein the pharmaceutical composition is composition MLC1501.

[0125] Embodiment 37: The herbal composition for use according to any one of embodiments 29 to 36, wherein the effective amount of the herbal composition is from about 1 mg / kg to about 100 mg / kg. Embodiment 38: The herbal composition for use according to any one of embodiments 29 to 37, wherein the method comprises administering a second anti-inflammatory agent.

[0126] Embodiment 39: The herbal composition for use according to any one of embodiments 29 to 38, wherein the injury results from chemical, mechanical or surgical intervention for treating the underlying disease or disorder according to any one of embodiments 21 to 24.

[0127] Embodiment 40: A herbal composition comprising at least four herbal ingredients, Radix Polygalae (Primula Root), Radix Astragali (Astragalus Root), Rhizome Ligusticum Chuanxiong, and Radix Angelica sinensis (Angelica Root), or extracts thereof, for use in treating a subject at risk of developing a disease or condition as a result of neuroinflammation, to prevent or alleviate the onset of the disease or condition.

[0128] Embodiment 41: The herbal composition for use according to embodiment 40, wherein the herbal composition further comprises at least one herbal ingredient selected from the group consisting of Radix et Rhizome Salviae Miltiorrhizae (red sage root), Radix Paeoniae Rubra (red peony root), Carthamus Tinctorius (safflower) flower, Semen Persicae (peach (Prunus Persica) seed), and Rhizome Acori Tatarinowii (iris rhizome) or extracts thereof.

[0129] Embodiment 42: The herbal composition for use according to embodiment 40, wherein the herbal composition consists essentially of at least four herbal ingredients, Polygalae Root (Radix Polygalae), Astragali Root (Radix Astragali), Rhizome Ligusticum Chuanxiong, and Angelica Root (Radix Angelica sinensis) or extracts thereof.

[0130] Embodiment 43: The herbal composition for use according to embodiment 40, wherein the herbal composition consists essentially of at least nine herbal ingredients, Polygala Root (Radix Polygalae) (Root of Japanese Bush Clovers), Astragalus Root (Radix Astragali) (Root of Astragalus Root), Cnidium Rhizome (Rhizome Ligusticum Chuanxiong), Angelica Root (Radix Angelica sinensis) (Root of Chinese Angelica), Salvia Root and Rhizome (Radix et Rhizome Salviae Miltiorrhizae) (Red Sage Root), Radix Paeoniae Rubra (Red Peony Root), Carthamus Tinctorius (Safflower) Flower, Peach Seed (Semen Persicae) (Peach (Prunus Persica) Seed), and Rhizome Acori Tatarinowii (Rhizome of Japanese Calamus) or extracts thereof.

[0131] Embodiment 44: The herbal composition for use according to any one of embodiments 40 to 43, wherein the herbal composition is a pharmaceutical composition. Embodiment 45: The herbal composition for use according to embodiment 44, wherein the pharmaceutical composition also comprises a pharmaceutically acceptable carrier or excipient.

[0132] Embodiment 46: The herbal composition for use according to embodiment 45, wherein the pharmaceutical composition is composition MLC901. Embodiment 47: The herbal composition for use according to embodiment 45, wherein the pharmaceutical composition is composition MLC1501.

[0133] Embodiment 48: The herbal composition for use according to any one of embodiments 40 to 47, wherein the effective amount of the herbal composition is from about 1 mg / kg to about 100 mg / kg. Embodiment 49: The herbal composition for use according to any one of embodiments 1 to 48, wherein the subject has a brain injury.

[0134] Embodiment 50: The herbal composition for use according to any one of embodiments 1 to 48, wherein the subject has brain swelling. Embodiment 51: The herbal composition for use according to any one of embodiments 1 to 48, wherein the subject has a leaky blood-brain barrier.

[0135] Embodiment 52: The herbal composition for use according to any one of embodiments 1 to 48, wherein the subject has had a stroke. Embodiment 53: The herbal composition for use according to any one of embodiments 1 to 48, wherein the subject has a neuropsychiatric disorder (e.g., depression, stress, post-traumatic stress disorder, schizophrenia, autism spectrum disorder or epilepsy).

[0136] Embodiment 54: A herbal composition comprising Radix Polygalae (Primitive bush clover root), Radix Astragali (Astragalus root), Rhizome Ligusticum Chuanxiong, and Radix Angelica sinensis (Angelica root) or extracts thereof for use in treating neuroinflammation in a subject with diabetes mellitus.

[0137] Embodiment 55: A herbal composition comprising Polygalae Root (Radix Polygalae) (Root of Japanese Bush Clover), Astragali Root (Root of Astragali Root), Cnidium Rhizome (Rhizome Ligusticum Chuanxiong), Angelica Sinensis Root (Root of Chinese Angelica), Salviae Miltiorrhizae Root and Rhizome (Red Sage Root), Paeoniae Rubra Root (Red Peony Root), Carthamus Tinctorius (Safflower) Flower, Peach Seed (Semen Persicae) (Peach (Prunus Persica) Seed), and Rhizome Acori Tatarinowii (Rhizome of Japanese Calamus) or extracts thereof for use in treating neuroinflammation in a subject with diabetes mellitus.

[0138] Embodiment 56: The herbal composition for use according to embodiment 54, wherein the herbal composition consists essentially of Radix Polygalae (Primitive bush clover root), Radix Astragali (Astragalus root), Rhizome Ligusticum Chuanxiong, and Radix Angelica sinensis (Angelica root) or extracts thereof.

[0139] Embodiment 57: The herbal composition for use according to embodiment 55, wherein the herbal composition consists essentially of Polygalae root (Radix Polygalae) (root of Japanese bush clover), Astragalus membranaceus root (Radix Astragali) (root of Astragali), Cnidium rhizome (Rhizome Ligusticum Chuanxiong), Angelica sinensis root (Radix Angelica sinensis) (root of Chinese angelica), Salvia root and rhizome (Radix et Rhizome Salviae Miltiorrhizae) (red sage root), Radix Paeoniae Rubra (red peony root), Carthamus Tinctorius (safflower) flower, Peach seed (Semen Persicae) (peach (Prunus Persica) seed), and Rhizome Acori Tatarinowii (rhizome of Aconite) or extracts thereof.

[0140] Embodiment 58: The herbal composition for use according to any one of embodiments 54 to 57, wherein the herbal composition is a pharmaceutical composition. Embodiment 59: The herbal composition for use according to embodiment 58, wherein the pharmaceutical composition also comprises a pharmaceutically acceptable carrier or excipient.

[0141] Embodiment 60: The herbal composition for use according to embodiment 58, wherein the pharmaceutical composition is composition MLC901. Embodiment 61: The herbal composition for use according to embodiment 58, wherein the pharmaceutical composition is composition MLC1501.

[0142] Embodiment 62: The herbal composition for use according to any one of embodiments 54 to 61, wherein the effective amount of the herbal composition is from about 1 mg / kg to about 100 mg / kg. Embodiment 63: The herbal composition for use according to any one of embodiments 54 to 62, wherein the method comprises administering a second anti-inflammatory agent.

[0143] Embodiment 64: The herbal composition for use according to any one of embodiments 54 to 63, wherein the subject is tested for and confirmed to have neuroinflammation prior to administering the herbal composition.

[0144] Embodiment 65: The herbal composition for use according to embodiment 64, wherein testing shows that the subject presents high levels of pro-inflammatory chemokines and / or cytokines. Embodiment 66: The herbal composition for use according to embodiment 64, wherein testing shows that the subject presents elevated levels of one or more pro-inflammatory chemokines selected from IL-1β, IL-6 and TNF-α and / or elevated levels of one or more inflammasomes selected from NLRP3 and iNOS.

[0145] Embodiment 67: The herbal composition for use according to any one of embodiments 54 to 66, wherein the neuroinflammation is caused by brain injury. Embodiment 68: The herbal composition for use according to any one of embodiments 54 to 66, wherein the neuroinflammation is caused by brain swelling.

[0146] Embodiment 69: The herbal composition for use according to any one of embodiments 54 to 66, wherein the neuroinflammation is caused by leakage of the blood-brain barrier. Embodiment 70: The herbal composition for use according to any one of embodiments 54 to 66, wherein the neuroinflammation is caused by stroke.

[0147] Embodiment 71: The herbal composition for use according to any one of embodiments 54 to 66, wherein the neuroinflammation is caused by a neuropsychiatric disorder (e.g., depression, stress, post-traumatic stress disorder, schizophrenia, autism spectrum disorder or epilepsy).

[0148] The following example describes a study to investigate the effect of MLC1501 on inflammation, and more specifically, its effect on the expression of various cellular markers involved in neuroinflammation. The study was divided into two parts: 1 / Characterization of the effects of MLC1501 after LPS-induced inflammation, focusing on the in vitro expression of the markers IL-1β, IL-6, TNF-α, NLRP3 and iNOS.

[0149] 2 / Characterization of the effects of MLC1501 in a model of focal cerebral ischemia, focusing on the in vivo expression of the pro-inflammatory cytokines IL-1β and IL-6. The results obtained below for MLC1501 may also be achieved with other herbal compositions of the present disclosure, including MLC901. [Example]

[0150] Example 1 1) Materials and Methods a) Cell culture The BV-2 microglial cell line was derived from neonatal mouse microglia immortalized with the v-raf / v-myc oncogene (Blasi et al., 1990). It was cultured in DMEM "high glucose" medium supplemented with 5% fetal calf serum (FCS) and 1% penicillin / streptomycin (Gibco, Thermofisher Scientific, USA). Cells were maintained in a humidified incubator at 37°C in an atmosphere containing 5% CO2. As soon as the cells reached 80% confluence, they were used for experiments.

[0151] b) In vitro treatment Cells were pretreated with MLC1501 at a concentration of 250 μg / mL for 24 hours. Eight different concentrations of MLC1501 were selected for dose-response analysis: 0, 10, 50, 100, 250, 500, 1000, and 2000 μg / mL.

[0152] c) In vitro inflammation model: LPS stimulation Cells are stimulated with 100 ng / mL of LPS (E. coli; O111:B4, Sigma-Aldrich) for 6 h in the absence of SVF but in the presence of MLC1501 (250 μg / mL). The experimental protocol is shown in Figure 1.

[0153] d)Animals Seven-week-old male C57Bl / 6JRj mice (at the time of receipt) were housed five per cage in an animal house maintained at an ambient temperature of 21±1° C. Animals had free access to food and drink.

[0154] e) In vivo treatment A single dose of 40 μg / kg MLC1501 (100 μL bolus) diluted in saline (vehicle) was injected intraperitoneally 60 min before the onset of ischemia. A schematic diagram of the protocol is shown in Figure 2.

[0155] f) MCAO model of ischemic stroke A 60-minute focal ischemia model was performed on vehicle- and MLC1501-treated mice. The model consisted of occlusion of the right middle cerebral artery (MCAO) with a silicone-coated 6-0 filament (Doccol, Redlands, CA, USA) for 60 minutes, followed by reperfusion (or, in some cases, no reperfusion). Cerebral blood flow within the region of interest was monitored by a laser Doppler (Perimed, Craponne, France) to control the severity of the pattern and the presence of reperfusion. Animals with a reduction in cerebral blood flow of less than 70% were excluded from the study (<1%). Sham surgery was performed on a control group of animals (insertion of a filament into the carotid artery without causing occlusion of the MCA). The animals were then housed in a thermostatically controlled recovery chamber (30–31°C) supplied with synthetic air for 30 hours.

[0156] g) mRNA extraction and quantitative rt-PCR Microglial cells and the ipsilateral brain hemisphere were collected 30 hours after the initiation of each in vitro and in vivo protocol. mRNA from microglial cells and brain tissue was extracted with RNAzol (Euromedex, France) and then reverse transcribed using Superscript IV (Thermofisher Scientific, USA). Expression of mRNA encoding IL-1β, IL-6, and TNF-α genes was then determined by qPCR (SybrGreen 480 Roche) using specific primers (Eurogentec, Angers, France) (sequences detailed in Figure 3). Results were normalized by the expression of housekeeping genes, GAPDH, or RPL0. Values ​​are expressed as 2-ΔΔCT normalized by the control. Duplicates were performed for each sample.

[0157] h) Statistics and Regression Data are presented as mean ± SEM, with each sample being the average of two replicates. Regression was performed using GraphPad Prism software. Outliers were determined by the ROUT method (Q = 1%). Statistical analysis between different animal groups was calculated using the Mann-Whitney test. The significance level was set at P < 0.05.

[0158] 2) Results a) MLC1501 reduces the relative expression of genes involved in the pro-inflammatory response in an in vitro inflammation model: Characterization of the anti-inflammatory effects of MLC1501 at a concentration of 250 μg / mL on BV2 microglial cell line after 1 / LPS stimulation We observed that 6 hours of LPS stimulation in cells not treated with MLC1501 resulted in a strong increase in the expression levels of mRNA encoding the pro-inflammatory cytokine genes IL-1β, IL-6, and TNF-α. The same LPS stimulation also induced increased expression of the NLRP3 and iNOS genes in untreated cells (Figure 4). Administration of MLC1501 at a concentration of 250 μg / mL resulted in a significant decrease in the relative expression of IL-1β, IL-6, TNF-α, NLRP3, and iNOS genes 6 hours after LPS stimulation compared with vehicle administration (Figure 4).

[0159] Dose-response analysis of MLC1501 on the expression of two / three inflammatory markers: IL-1β, IL-6, and TNF-α Eight different concentrations of MLC1501 were selected for dose-response analysis: 0, 10, 50, 100, 250, 500, 1000, and 2000 μg / mL. Administration of MLC1501 completely reduced the relative overexpression of IL-1β, IL-6, and TNF-α in a dose-dependent manner in the presence of LPS. According to the regression analysis, the IC50 (median inhibitory concentration) values ​​for IL-1β, IL-6, and TNF-α were 41.28 ± 1.50 μg / mL, 93.43 ± 2.61 μg / mL, and 34.43 ± 1.64 μg / mL, respectively (Figure 5).

[0160] (b) MLC1501 reduced the relative expression of pro-inflammatory genes in an in vivo focal ischemia model: MCAO-induced focal ischemia in the vehicle group induced high expression of pro-inflammatory genes. Administration of MLC1501 60 min before the onset of ischemia resulted in a significant decrease in the relative expression of genes encoding the cytokines IL-1β and IL-6 (Figure 6).

[0161] 3) Conclusion This study demonstrated that administration of MLC1501 at a concentration of 250 μg / mL resulted in a reduction in inflammation by decreasing the expression of mRNA encoding genes involved in harmful inflammation: IL-1β, IL-6, TNF-α, NLRP3, and iNOS. This phenomenon was observed both in vitro after LPS stimulation and in vivo after focal cerebral ischemia.

[0162] All references, articles, publications, patents, patent publications, and patent applications cited herein are incorporated by reference in their entirety for all purposes. However, the mention of any reference, article, publication, patent, patent publication, or patent application herein is not, and should not be construed as, an acknowledgment or any form of suggestion that they constitute valid prior art or form part of the common general knowledge in any country in the world.

Claims

1. 1. A method for treating neuroinflammation in a subject, comprising administering to the subject an effective amount of an herbal composition comprising Radix Polygalae (Philippine bush clover root), Radix Astragali (Astragalus root), Rhizome Ligusticum Chuanxiong (Cnidium rhizome), and Radix Angelica sinensis (Angelica sinensis root), or extracts thereof.

2. 1. A method of treating neuroinflammation in a subject, comprising administering to a subject a compound selected from the group consisting of Polygala Root (Radix Polygalae) (root of bush clover), Astragalus Root (Radix Astragali) (root of astragalus), Cnidium Rhizome (Rhizome Ligusticum Chuanxiong), Angelica Root (Radix Angelica sinensis) (root of Chinese angelica), Salvia Root and Rhizome (Radix Salviae Miltiorrhizae) (red sage root), Radix Paeoniae Rubra (red peony root), and Carthamus tinctorius (safflower) root.

1. A method for treating neuroinflammation in a subject, comprising administering to said subject an effective amount of an herbal composition comprising flowers of Safflower (Safflower) (Salf Vulgare), Semen Persicae (Peach (Prunus Persica) seeds), and Rhizome Acorii Tatarinowii (Rhizome of the Calamus) or extracts thereof.

3. 1. A method for treating neuroinflammation in a subject, comprising administering to said subject an effective amount of an herbal composition consisting essentially of Radix Polygalae (Philippine bush clover root), Radix Astragali (Astragalus root), Rhizome Ligusticum Chuanxiong (Cnidium rhizome), and Radix Angelica sinensis (Angelica root), or extracts thereof.

4. 1. A method of treating neuroinflammation in a subject, comprising administering to a subject a compound selected from the group consisting of Polygala Root (Radix Polygalae) (root of bush clover), Astragalus Root (Radix Astragali) (root of astragalus), Cnidium Rhizome (Rhizome Ligusticum Chuanxiong), Angelica Root (Radix Angelica sinensis) (root of Chinese angelica), Salvia Root and Rhizome (Radix Salviae Miltiorrhizae) (red sage root), Radix Paeoniae Rubra (red peony root), and Carthamus tinctorius (safflower) root.

1. A method for treating neuroinflammation in a subject comprising administering to said subject an effective amount of an herbal composition consisting essentially of Safflower (Safflower) flowers (Safflower) (Semen Persicae) (Peach (Prunus Persica) seeds), and Rhizome Acorii Tatarinowii (Rhizome of the Calamus) or extracts thereof.

5. 1. A method for treating neuroinflammation in a subject with diabetes mellitus, comprising administering to the subject an effective amount of an herbal composition comprising Radix Polygalae (Philippine bush clover root), Radix Astragali (Astragalus root), Rhizome Ligusticum Chuanxiong (Cnidium rhizome), and Radix Angelica sinensis (Angelica sinensis root), or extracts thereof.

6. 1. A method for treating neuroinflammation in a subject with diabetes mellitus, comprising administering to a subject a compound selected from the group consisting of Polygala Root (Radix Polygalae) (root of bush clover), Astragalus Root (Radix Astragali) (root of astragalus), Cnidium Rhizome (Rhizome Ligusticum Chuanxiong), Angelica Root (Radix Angelica sinensis) (root of Chinese angelica), Salvia Root and Rhizome (Radix Salviae Miltiorrhizae) (red sage root), Radix Paeoniae Rubra (red peony root), and Carthamus tinctorius (safflower) root.

1. A method for treating neuroinflammation in a subject with diabetes mellitus, comprising administering to said subject an effective amount of an herbal composition comprising Safflower (Safflower) Flower (Sesame) (Peach) Seed (Semen Persicae) (Peach (Prunus Persica) Seed), and Rhizome Acorii Tatarinowii (Rhizome of the Calamus) or extracts thereof.

7. 1. A method for treating neuroinflammation in a subject with diabetes mellitus, comprising administering to said subject an effective amount of an herbal composition consisting essentially of Radix Polygalae (Philippine bush clover root), Radix Astragali (Astragalus root), Rhizome Ligusticum Chuanxiong (Cnidium rhizome), and Radix Angelica sinensis (Angelica root), or extracts thereof.

8. 1. A method for treating neuroinflammation in a subject with diabetes mellitus, comprising administering to a subject a compound selected from the group consisting of Polygala Root (Radix Polygalae) (root of bush clover), Astragalus Root (Radix Astragali) (root of astragalus), Cnidium Rhizome (Rhizome Ligusticum Chuanxiong), Angelica Root (Radix Angelica sinensis) (root of Chinese angelica), Salvia Root and Rhizome (Radix Salviae Miltiorrhizae) (red sage root), Radix Paeoniae Rubra (red peony root), and Carthamus tinctorius (safflower) root.

1. A method for treating neuroinflammation in a subject with diabetes mellitus, comprising administering to said subject an effective amount of an herbal composition consisting essentially of Safflower (Safflower) flowers (Safflower) (Semen Persicae) (Peach (Prunus Persica) seeds), and Rhizome Acorii Tatarinowii (Rhizome of the Calamus) or extracts thereof.

9. The method of any one of claims 1 to 8, wherein the neuroinflammation is the result of an injury.

10. 10. The method of claim 9, wherein the injury is selected from the group consisting of spinal cord injury, stroke, traumatic brain injury, subarachnoid hemorrhage, subdural hemorrhage, or epidural hemorrhage.

11. The method of any one of claims 1 to 8, wherein the neuroinflammation is the result of an infection.

12. 12. The method of claim 11, wherein the infection is an infection of the brain or central nervous system (e.g., encephalitis or bacterial, viral, or fungal meningitis).

13. The infectious disease may be a viral infection [e.g., varicella-zoster virus (VZV) infection, herpes simplex virus (HSV-1 or HSV-2), SARS-CoV-2 virus or a variant thereof, Epstein-Barr virus, hepatitis B virus, hepatitis C virus, human immunodeficiency virus type 1 (HIV-1), human papillomavirus (HPV), human T-cell lymphotropic virus type 1 (HTLV-1), or Kaposi's sarcoma herpesvirus (KSHV)], a fungal infection [e.g., a fungus such as Cryptococcus neoformans, Candida albicans, or Histoplasma capsulatum], or a fungal infection [e.g., a fungus such as Cryptococcus neoformans, Candida albicans, or Histoplasma capsulatum]. capsulatum, Coccidioides immitis, Paracoccidioides brasiliensis, Aspergillus spp., and zygomycetes], bacterial infections (e.g., the bacteria is selected from the group consisting of Listeria monocytogenes, Borrelia burgdorferi, Neisseria meningitidis, Streptococcus pneumoniae, Staphylococcus aureus, aureus, Haemophilus influenzae, Mycobacterium and Brucella species] or a parasitic infection.

14. The method of any one of claims 1 to 8, wherein the neuroinflammation is the result of exposure to a toxin.

15. The method of any one of claims 1 to 8, wherein the neuroinflammation is associated with a neurodegenerative disease (e.g., Alzheimer's disease, Parkinson's disease, Huntington's disease, frontotemporal dementia, amyotrophic lateral sclerosis, multiple sclerosis, 10-12 primary tauopathy, synucleinopathy (i.e., dementia with Lewy bodies and multiple system atrophy), polyglutamine diseases including spinocerebellar ataxia, prion diseases, traumatic brain injury, chronic traumatic encephalopathy, stroke, and spinal cord injury).

16. 9. The method of any one of claims 1 to 8, wherein the neuroinflammation is associated with a neuropsychiatric condition (e.g., depression, stress, post-traumatic stress disorder, schizophrenia, autism spectrum disorder, or epilepsy).

17. The method of any one of claims 1 to 8, wherein the neuroinflammation is associated with aging (e.g., "inflammagenesis").

18. 9. The method of any one of claims 1 to 8, wherein the neuroinflammation is associated with a disease or disorder selected from the group consisting of postoperative cognitive dysfunction, ischemia, brain tumor, chronic pain, headache, migraine, trigeminal neuralgia, cardiovascular disease and associated risk factors (e.g., due to obesity, hypertension, diabetes, hyperlipidemia, smoking, etc.), chronic peripheral inflammatory conditions (e.g., rheumatoid arthritis, inflammatory bowel disease, psoriasis, etc.), perturbation of the gut microbiota (i.e., dysbiosis), and herpes keratitis.

19. 9. The method of treatment of any one of claims 1 to 8, wherein the neuroinflammation results from an underlying disease or chemical intervention to treat a disease (e.g., the use of TNF inhibitors to treat autoimmune diseases such as rheumatoid arthritis).

20. The method of any one of claims 1 to 8, wherein the neuroinflammation results from an underlying disease or mechanical intervention to treat the disease.

21. The method of any one of claims 1 to 8, wherein the neuroinflammation results from an underlying disease or surgical intervention to treat a disease.

22. 22. The method of treatment of claim 21, wherein the surgical intervention is selected from the group consisting of carotid endarterectomy, carotid angioplasty with or without stenting, vertebral angioplasty with or without stenting, intracranial angioplasty with or without stenting, aneurysm coiling, occlusion of vascular malformations, repair of intracranial fistulas, thrombectomy, extracranial-intracranial bypass, open heart surgery, cardiac surgery requiring bypass surgery, coronary artery bypass grafting, coronary angioplasty with or without stenting, valve surgery / valvuloplasty, valvulotomy, closure of patent foramen ovale, and surgery on the great vessels of the heart.

23. A method of treatment according to any one of claims 1 to 22, wherein the herbal composition is a pharmaceutical composition.

24. 24. The method of claim 23, wherein the pharmaceutical composition also comprises a pharmaceutically acceptable carrier or excipient.

25. 24. The method of claim 23, wherein the pharmaceutical composition is composition MLC901.

26. 24. The method of claim 23, wherein the pharmaceutical composition is composition MLC1501.

27. 27. The method of treatment according to any one of claims 1 to 26, wherein the effective amount of the herbal composition is from about 1 mg / kg to about 100 mg / kg.

28. 28. The method of treatment of any one of claims 1 to 27, wherein the method comprises administering a second anti-inflammatory agent in addition to the herbal composition.

29. 23. The method of treatment according to any one of claims 19 to 22, wherein the herbal composition is administered before, simultaneously with, or after the administration of an intervention to treat the underlying disease or disorder.

30. 30. The method of treatment of any one of claims 1 to 29, wherein the subject is tested for and confirmed to have neuroinflammation prior to administering the herbal composition.

31. 31. The method of treatment of claim 30, wherein the test indicates that the subject exhibits high levels of pro-inflammatory chemokines and / or cytokines.

32. 31. The method of treatment of claim 30, wherein the test indicates that the subject exhibits elevated levels of one or more pro-inflammatory chemokines selected from IL-1β, IL-6, and TNF-α and / or elevated levels of one or more inflammasomes selected from NLRP3 and iNOS.

33. 1. A method of preventing or limiting neuroinflammation in a subject at risk of neuroinflammation as a result of injury, comprising administering to the subject an effective amount of an herbal composition comprising at least four herbal ingredients: Radix Polygalae (Philippine bush clover root), Radix Astragali (Astragalus root), Rhizome Ligusticum Chuanxiong (Cnidium rhizome), and Radix Angelica sinensis (Angelica sinensis root), or extracts thereof.

34. 34. The method of claim 33, wherein the herbal composition further comprises at least one herbal ingredient selected from the group consisting of Radix et Rhizome Salviae Miltiorrhizae (red sage root), Radix Paeoniae Rubra (red peony root), Carthamus Tinctorius (safflower), Semen Persicae (peach seed), and Rhizome Acorii Tatarinowii (calamus rhizome), or an extract thereof.

35. 34. The method of claim 33, wherein the herbal composition consists essentially of at least four herbal ingredients: Polygala Root (Radix Polygalae) (Root of Queen Elizabeth II), Astragalus Root (Radix Astragali), Cnidium Rhizome (Rhizome Ligusticum Chuanxiong), and Angelica Root (Radix Angelica sinensis) (Root of Angelica Root), or extracts thereof.

36. The herbal composition comprises at least nine herbal ingredients, Polygala Root (Radix Polygalae) (Hakubai Root), Astragalus Root (Radix Astragali) (Astragalus Root), Cnidium Rhizome (Rhizome Ligusticum Chuanxiong), Angelica Root (Radix Angelica sinensis) (Angelica Root), Salvia Root and Rhizome (Radix et Rhizome Salviae Miltiorrhizae) (Red Sage Root), Radix Paeoniae Rubra (Red Peony Root), Safflower (Carthamus tinctorius) and 34. The method of claim 33, wherein the active ingredient consists essentially of safflower (Safflower) flowers (Safflower) (Salvia tinctorius), peach seeds (Semper persicae) (Prunus persica) and Rhizome acorii tatarinowii (Rhizome of the Japanese iris) or extracts thereof.

37. The method according to any one of claims 33 to 36, wherein the herbal composition is a pharmaceutical composition.

38. 38. The method of claim 37, wherein the pharmaceutical composition also comprises a pharmaceutically acceptable carrier or excipient.

39. 38. The method of claim 37, wherein the pharmaceutical composition is composition MLC901.

40. 38. The method of claim 37, wherein the pharmaceutical composition is composition MLC1501.

41. 41. The method according to any one of claims 33 to 40, wherein the effective amount of the herbal composition is from about 1 mg / kg to about 100 mg / kg.

42. 42. The method of any one of claims 33 to 41, wherein the method comprises administering a second anti-inflammatory agent in addition to the herbal composition.

43. 43. The method of any one of claims 33 to 42, wherein the injury results from chemical, mechanical or surgical intervention to treat the underlying disease or disorder of any one of claims 19 to 22.

44. 1. A method for treating a subject at risk of developing a disease or condition as a result of neuroinflammation, comprising administering to the subject an effective amount of an herbal composition comprising at least four herbal ingredients: Radix Polygalae (Philippine bush clover root), Radix Astragali (Astragalus root), Rhizome Ligusticum Chuanxiong (Cnidium rhizome), and Radix Angelica sinensis (Angelica sinensis root), or extracts thereof, to prevent or alleviate the onset of the disease or condition.

45. 45. The method of claim 44, wherein the herbal composition further comprises at least one herbal ingredient selected from the group consisting of Radix et Rhizome Salviae Miltiorrhizae (red sage root), Radix Paeoniae Rubra (red peony root), Carthamus Tinctorius (safflower) flower, Semen Persicae (peach (Prunus Persica) seed), and Rhizome Acorii Tatarinowii (Rhizome of the Japanese Calamus) or extracts thereof.

46. 45. The method of claim 44, wherein the herbal composition consists essentially of at least four herbal ingredients: Radix Polygalae (Philippine bush clover root), Radix Astragali (Astragalus root), Rhizome Ligusticum Chuanxiong (Cnidium rhizome), and Radix Angelica sinensis (Angelica sinensis root), or extracts thereof.

47. The herbal composition comprises at least nine herbal ingredients, Polygala Root (Radix Polygalae) (Hakubai Root), Astragalus Root (Radix Astragali) (Astragalus Root), Cnidium Rhizome (Rhizome Ligusticum Chuanxiong), Angelica Root (Radix Angelica sinensis) (Angelica Root), Salvia Root and Rhizome (Radix et Rhizome Salviae Miltiorrhizae) (Red Sage Root), Radix Paeoniae Rubra (Red Peony Root), Safflower (Carthamus tinctorius) and 45. The method of claim 44, wherein the active ingredient consists essentially of Safflower (Safflower) flowers (Safflower) (Semen Persicae) (Peach (Prunus Persica) seeds), and Rhizome Acorii Tatarinowii (Rhizome of the Calamus) or extracts thereof.

48. A method of treatment according to any one of claims 44 to 47, wherein the herbal composition is a pharmaceutical composition.

49. 49. The method of treatment of claim 48, wherein the pharmaceutical composition also comprises a pharmaceutically acceptable carrier or excipient.

50. 49. The method of treatment of claim 48, wherein the pharmaceutical composition is composition MLC901.

51. 49. The method of claim 48, wherein the pharmaceutical composition is composition MLC1501.

52. 52. The method of treatment according to any one of claims 44 to 51, wherein the effective amount of the herbal composition is from about 1 mg / kg to about 100 mg / kg.

53. 53. The method of treatment according to any one of claims 44 to 52, wherein the disease or condition is brain injury.

54. 53. The method of any one of claims 44 to 52, wherein the disease or condition is brain swelling.

55. 53. The method of treatment of any one of claims 44 to 52, wherein the disease or condition is a leakage of the blood-brain barrier.

56. 53. The method of treatment according to any one of claims 44 to 52, wherein the disease or condition is stroke.

57. 53. The method of any one of claims 44 to 52, wherein the disease or condition is a neuropsychiatric disorder (e.g., depression, stress, post-traumatic stress disorder, schizophrenia, autism spectrum disorder, or epilepsy).

58. 58. The method of treatment according to any one of claims 1 to 57, comprising administering to the subject an effective amount of the herbal composition prior to injury.

59. 59. The method of treatment of claim 58, wherein the herbal composition is composition MLC901.

60. 59. The method of treatment of claim 58, wherein the herbal composition is composition MLC1501.

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