Treatment of inflammation

Abstract

The present disclosure relates to a method of treating neuroinflammation. In one aspect, the disclosure provides for administering to a subject suffering from neuroinflammation an effective amount of at least four herbal components: polygala tenuifolia, astragalus membranaceus, ligusticum wallichii, and angelica sinensis, roots or rhizomes thereof, or extracts thereof.

Description

Technical Field

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

[0002] Neuroinflammation is inflammation of neural tissues such as the brain and spinal cord. Neuroinflammation is a response of the CNS innate immune system that is triggered following insults caused, for example, by injury, infection, exposure to toxins, neurodegenerative disease, aging (e.g., "inflammaging"), or by chemical, mechanical, or surgical intervention to treat the underlying condition or disease.

[0003] Neuroinflammation can be mediated by the production of reactive oxygen species, chemokines and cytokines. These mediators are produced by CNS glial cells, including microglia and astrocytes, as well as endothelial cells and immune cells of peripheral origin. The role of microglia during systemic inflammation is similar to that of macrophages, and is considered to be an important cell type involved in neuroinflammation. In response to damage, microglia can be converted into different functional states, thereby changing its proliferation, morphology (i.e., shortened process), phagocytic activity (clearing foreign organisms and cell debris) and antigen presentation. However, in addition to these beneficial processes in response to damage, microglia also release proinflammatory factors, such as cytokines, chemokines and matrix metalloproteinases (MMPs), which can damage the function of the blood-brain barrier (BBB). In general, the multiphase (beneficial and harmful) effects of neuroinflammation are time-dependent, but the long-term activation of microglia can cause damage to brain tissue and BBB.

[0004] There is currently a lack of effective and safe therapeutic agents that can modulate neuroinflammation to limit or prevent the negative effects of neuroinflammation on brain tissue and the BBB. Therefore, new therapeutic approaches to modulate neuroinflammation are urgently needed. Summary of the invention

[0005] The present disclosure provides a combination of a traditional Chinese herbal medicine product or an extract thereof, wherein the combination reduces the activation of microglia after damage and the secretion of inflammatory cytokines and other neuroinflammatory mediators. Therefore, the combination is particularly effective in preventing and / or controlling neuroinflammation, treating, reducing or preventing symptoms associated with excessive microglia activation, and alleviating the immune, physiological, biochemical and psychological consequences of neuroinflammation (including damage to brain tissue and BBB).

[0006] Thus, according to a first aspect, there is provided a method of treating neuroinflammation in a subject in need thereof, the method comprising administering to the subject an effective amount of an herbal composition comprising at least four herbal components: Polygalae (thin leaf milkwort), Astragali (membranous milkvetch), Ligusticum Chuanxiong (Ligusticum Chuanxiong) and Angelica sinensis (Chinese angelica), their roots or rhizomes, or extracts thereof. As used in this aspect and 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 aspects, the herbal composition comprising four herbal components: Polygala tenuifolia, Astragalus membranaceus, Ligusticum chuanxiong and Angelica sinensis, their roots or rhizomes, or extracts thereof further comprises at least one herbal component selected from the group consisting of Salviae Miltiorrhizae (red sage), Paeoniae Rubra (red peony), flower of Carthamus Tinctorius (safflower), Semen Persicae (peach seed) and Acori Tatarinowii (grassleaf sweetflag), their roots or rhizomes, or extracts thereof.

[0008] In a further aspect, a method of treating neuroinflammation in a subject in need thereof is provided, the method comprising administering to the subject an effective amount of an herbal composition consisting essentially of at least four herbal components: Polygala tenuifolia, Astragalus membranaceus, Ligusticum chuanxiong, Angelica sinensis, their roots or rhizomes, or extracts thereof.

[0009] In another aspect, a method of treating neuroinflammation in a subject in need thereof is provided, the method comprising administering to the subject an effective amount of an herbal composition consisting essentially of at least nine herbal components: Polygala tenuifolia, Astragalus membranaceus, Ligusticum chuanxiong, Angelica sinensis, Salvia miltiorrhiza, Paeonia lactiflora, Carthamus tinctorius flower, Persica persica (peach seed), and Acorus tatarinowii, 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 further comprise one or more pharmaceutically acceptable carriers or excipients.

[0011] In one embodiment of any of the above aspects, the neuroinflammation is associated with an injury, such as a spinal cord injury, a stroke, a traumatic brain injury, a subarachnoid hemorrhage, a subdural hemorrhage, or an epidural hemorrhage.

[0012] In one embodiment of any of the above aspects, 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 abscess. Examples of viral infections include varicella zoster virus (VZV) infection, herpes simplex virus (HSV-1 or HSV-2), SARS-CoV-2 virus or variants thereof, Epstein-Barr virus, hepatitis B virus, hepatitis C virus, human immunodeficiency virus type 1 (HIV-1), human papillomavirus (HPV), human T-lymphotropic virus type 1 (HTLV-1), and Kaposi's sarcoma herpesvirus (KSHV). Exemplary fungi that can cause neuroinflammation include Cryptococcus neoformans, Candida albicans, Histoplasma capsulatum, Coccidioides immitis, Paracoccidioides brasiliensis, Aspergillus species and zygomycetes. Examples of antibacterials that can cause neuroinflammation include Listeria monocytogenes, Borrelia burgdorferi, Neisseria meningitidis, Streptococcus pneumoniae, Staphylococcus aureus, Haemophilus influenzae, Mycobacterium and Brucella species. Neuroinflammation may also be associated with parasitic infections.

[0013] In one embodiment of any of the above aspects, the neuroinflammation is associated with exposure to a toxin.

[0014] In one embodiment of any of the above aspects, neuroinflammation is associated with a neurodegenerative disease such as Alzheimer's disease, Parkinson's disease, Huntington's disease, frontotemporal dementia, amyotrophic lateral sclerosis, multiple sclerosis, 10-12 primary tauopathy, synucleinopathies (i.e., Lewy body dementia and multiple system atrophy), polyglutamine diseases including spinocerebellar ataxia, prion diseases, traumatic brain injury, chronic traumatic encephalopathy, stroke, and spinal cord injury.

[0015] In one embodiment of any of the above aspects, neuroinflammation is associated with a neuropsychiatric disorder, such as depression, stress, post-traumatic stress disorder, schizophrenia, autism spectrum disorder, or epilepsy.

[0016] In one embodiment of any of the above aspects, neuroinflammation is associated with aging (eg, "inflammaging").

[0017] Other conditions that can lead to neuroinflammation include, for example, postoperative cognitive dysfunction, ischemia, brain tumors, chronic pain, headache, migraine, trigeminal neuralgia, cardiovascular disease and related risk factors (e.g. due to obesity, hypertension, diabetes, hyperlipidemia, smoking, etc.), chronic peripheral inflammatory disorders (e.g., rheumatoid arthritis, inflammatory bowel disease, psoriasis, etc.), intestinal microbiota disturbances (i.e., dysbiosis), and herpetic keratitis.

[0018] In one embodiment of any of the above aspects, the neuroinflammation is associated with a chemical intervention to treat the underlying condition or disease. An example of such a chemical intervention is the use of TNF inhibitors to treat autoimmune diseases such as rheumatoid arthritis.

[0019] In one embodiment of any of the above aspects, the neuroinflammation is associated with a mechanical intervention to treat the underlying condition or disease.

[0020] In one embodiment of any of the above aspects, the neuroinflammation is associated with a surgical intervention to treat the underlying condition or disease. Examples of surgical interventions include carotid endarterectomy, carotid angioplasty with or without stents, vertebral angioplasty with or without stents, intracranial angioplasty with or without stents, aneurysm coil embolization, vascular malformation occlusion, intracranial fistula repair, thrombectomy, extracranial-intracranial bypass, open heart surgery, heart surgery requiring a bypass machine, coronary artery bypass, coronary angioplasty with or without stents, valve surgery / valvuloplasty, valvulotomy, patent foramen ovale closure, and cardiac great vessel surgery.

[0021] In one embodiment of any of the above aspects, there is provided administration of an effective amount of the herbal composition, the effective amount being from about 1 mg / kg to about 100 mg / kg.

[0022] In one embodiment of any of the above aspects, a method is provided comprising administering a second agent for treating an underlying condition or disease along with the above herbal composition.

[0023] In one embodiment of any of the above aspects, there is provided administration of an effective amount of the herbal composition to control neuroinflammation, thereby promoting recovery after brain injury.

[0024] 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 brain injury.

[0025] 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, thereby facilitating neurovascular repair.

[0026] 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, thereby promoting recovery after brain injury.

[0027] 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 injury.

[0028] In specific embodiments of any of the above aspects and embodiments, neuroinflammation is treated or reduced in a subject with an underlying condition. In a specific embodiment, the underlying condition is diabetes. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] The accompanying drawings are included to illustrate specific embodiments of the present disclosure and related technologies. The accompanying drawings provide a further understanding of the present disclosure and are incorporated into and constitute a part of the present disclosure. It should be understood that the accompanying drawings illustrate embodiments of the present disclosure and together with the rest of the present disclosure explain the principles of the present disclosure. It should be clear that the accompanying drawings illustrate embodiments of the present disclosure and that a person of ordinary skill in the art can easily understand other embodiments from the accompanying drawings described herein.

[0030] Figure 1 shows a chronological diagram of the experimental protocol for in vitro inflammation studies.

[0031] Figure 2 shows a chronological diagram of the experimental protocol for the in vivo studies.

[0032] FIG. 3 is a table showing the sequences of qPCR (quantitative polymerase chain reaction) primers used.

[0033] Figure 4 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).

[0034] FIG5 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.

[0035] FIG6 shows the effect of MLC1501 on the relative expression of IL-1β and IL-6 after middle cerebral artery occlusion (MCAO)-induced cerebral ischemia.

[0036] Glossary

[0037] This section is intended to provide guidance on the interpretation of the words and phrases (and their grammatical variations where appropriate) set out below.

[0038] Unless otherwise stated, the terms "include" and "comprising" and grammatical variations thereof are intended to represent "open" or "inclusive" language such that the listed elements are included but also permit the inclusion of additional, non-listed elements.

[0039] As used herein, the term "about" in relation to a numerical value means, for example, about ±30%, about ±20%, about ±10%, about ±5% or about ±1% of the numerical value. If necessary, the word "about" may be omitted from the definition of the disclosure herein.

[0040] The words "a", "an" and "the" are used to describe elements and components of the present invention. This is done for convenience only and to give a general sense of the present invention. Unless the context clearly indicates otherwise, the description should be understood to include one or at least one, and the singular also includes the plural. Thus, for example, the term "agent" includes reference to a single agent as well as multiple agents (including mixtures of agents). It should also be noted that the term "or" is generally used in its sense that includes "and / or" unless the context clearly indicates otherwise.

[0041] As used herein, the term "inflammaging" refers to the age-related increase in pro-inflammatory markers in blood and / or tissues, which may contribute to the pathogenesis of age-related diseases.

[0042] As used herein, the term "in vivo" includes reference to the use of a whole living organism. This is in contrast to the term "in vitro", in which a whole living organism is not used. The term "in vitro" is to be understood to include, inter alia, "ex vivo" uses, in which cells, tissues, etc. that do not form part of a whole living organism (e.g., cells or tissues from cell or tissue culture, biopsies, dead organisms, etc.) may be used. Further non-limiting examples of "in vitro" involve the use of cell extracts or lysates.

[0043] The terms "patient" and "subject" are used interchangeably herein, and these terms include reference to any human or non-human animal (preferably a mammal) that is desired to be treated using the present invention. However, it should be understood that "patient" or "subject" does not mean the presence of symptoms. As used herein, the term "mammal" refers to any member of the mammal class, including but not limited to humans and non-human primates, such as chimpanzees and other apes and monkey species; farm animals, such as cattle, sheep, pigs, goats and horses; domestic / companion animals, such as dogs and cats; experimental animals (e.g., rabbits and rodents, such as mice, rats and guinea pigs, etc.). Preferably, the mammal is a human.

[0044] The terms "treatment" and "treating" include any use to rescue a disease state or symptom, prevent the establishment of a disease, or otherwise prevent, hinder, delay or reverse the progression of a disease or other undesirable symptoms. Thus, "treatment" and "treating" include both prophylactic and therapeutic treatments. In the context of "a method for treating neuroinflammation" herein, "treatment" encompasses modulating neuroinflammation to limit or prevent the negative effects of neuroinflammation on brain tissue and the BBB, while limiting the impact on its beneficial effects.

[0045] Throughout this disclosure, certain embodiments may be disclosed in a range format. It should be understood that the description of the range format is only for convenience and brevity, and should not be interpreted as a rigid limitation on the scope of the disclosed range. Therefore, the description of the range should be considered to have specifically disclosed all possible sub-ranges and each numerical value within the range. For example, the description of the range, such as from 1 to 6 should be considered as a sub-range that has been specifically disclosed, such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6, etc., and a single number within the range, such as 1, 2, 3, 4, 5 and 6. This applies regardless of the breadth of the range.

[0046]

[00136] The terms "disease," "disorder," and "condition" are used interchangeably herein unless the context clearly dictates otherwise.

[0047] Unless defined otherwise, 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 belongs. DETAILED DESCRIPTION

[0048] With reference to the following drawings, detailed embodiments and implementations of the claimed subject matter are disclosed in detail in terms of technical content, structural features, and the purpose and effect of realization. It should be understood that the disclosed embodiments and implementations are merely illustrations of the claimed subject matter that can be embodied in various forms. However, the present disclosure can be embodied in many different forms and should not be construed as being limited to the exemplary embodiments and implementations set forth herein. On the contrary, these exemplary embodiments and implementations are provided to make the description of the present disclosure thorough and complete, and to fully convey the scope of the present disclosure to those skilled in the art. Specifically, the terms in the embodiments of the present disclosure are only used to describe the purpose of the present invention and are not used to limit the present disclosure. In the following description, the details of well-known features and technologies may be omitted to avoid unnecessarily obscuring the presented embodiments and implementations.

[0049] The configurations discussed in the following description are non-limiting examples that may vary and are merely recorded to illustrate at least one embodiment and are not intended to limit the scope thereof.

[0050] Microglia are the main resident macrophages of the central nervous system, and account for 5% to 12% of brain cells. Microglia act as cell effectors, and therefore play an important role in the immune response of the central nervous system. Therefore, microglia are essential to the defense and tissue repair process of the body under physiological conditions. However, the proinflammatory immune response driven by microglia is also a key factor in the pathogenesis of several brain diseases. The aggravated activation of microglia (for example, caused by LPS or by ischemia) can provide a release of chemokines and proinflammatory cytokines (such as IL-1 β, IL-6 and TNF-α) to induce harmful inflammatory responses. The release of this cytokine and chemokine can occur through various mediators.

[0051] A key step in the inflammatory response is the activation of inflammasomes, which are intracellular protein complexes that contain pattern recognition receptors and other inflammatory molecules. It is known that the secretion of IL-1β is directly mediated by the formation of inflammasomes, including NLRP3.

[0052] Another mediator of proinflammatory cytokine and chemokine secretion is iNOS. It can be induced by endotoxins (such as LPS) and cytokines. Its induction leads to the production of nitric oxide (NO), a free radical associated with cytotoxic function of microglia, apoptosis and oxidative stress.

[0053] In addition to causing neuronal death, these cytokines and proinflammatory mediators promote damage to the blood-brain barrier and, therefore, promote leukocyte infiltration into the brain.

[0054] Certain herbal compositions are identified herein that limit the release of proinflammatory cytokines, including the cytokines IL-1β, IL-6, and TNF-α. The herbal compositions herein also limit the formation of inflammasomes, including NLRP3 and iNOS. Therefore, the herbal compositions are particularly useful for treating inflammation associated with these cytokines and inflammasomes, including treating neuroinflammation.

[0055] Therefore, in one aspect of the present disclosure, a method for treating neuroinflammation in a subject in need thereof is provided, the method comprising administering to the subject an effective amount of an herbal composition comprising at least four herbal components: Polygala tenuifolia (root of Polygala), Astragalus membranaceus (root of Astragalus), Ligusticum chuanxiong rhizome and Angelica dahurica (root of Angelica sinensis), or extracts thereof.

[0056] In one embodiment of the above aspects, the herbal composition comprises four herbal components: Polygala root (root of Polygala), Astragalus root (root of Astragalus), Ligusticum chuanxiong rhizome and Angelica root (root of Angelica sinensis), and further comprises at least one herbal component selected from the group consisting of Salvia miltiorrhiza root and rhizome (root of Salvia miltiorrhiza), Paeonia rubra root (root of Paeonia rubra), Carthamus tinctorius flower, Prunus persica (peach seed) and Acorus calamus rhizome (rhizome of Acorus calamus), or extracts thereof.

[0057] In another aspect, a method of treating neuroinflammation in a subject in need thereof is provided, the method comprising administering to the subject an effective amount of an herbal composition consisting essentially of at least four herbal components: Polygala tenuifolia (root of Polygala), Astragalus membranaceus (root of Astragalus), Ligusticum chuanxiong rhizome, and Angelica dahurica (root of Angelica sinensis), or extracts thereof.

[0058] In another aspect, a method of treating neuroinflammation in a subject in need thereof is provided, the method comprising administering to the subject an effective amount of an herbal composition consisting essentially of at least nine herbal components: Polygala root (Polygala tenuifolia root), Astragalus root (Astragalus root), Ligusticum chuanxiong rhizome and Angelica acutiloba root (Angelica root), Salvia miltiorrhiza root and rhizome (Salvia miltiorrhiza root), Paeonia rubra root (Paeonia rubra root), Carthamus tinctorius flower, Prunus persica (Peach seed) and Acorus tatarinowii rhizome (Acorus calamus rhizome), or extracts thereof.

[0059] In one embodiment of any of the above aspects, neuroinflammation is treated or reduced in a subject with an underlying condition such as cardiovascular disease, diabetes, obesity, or atherosclerosis. In a specific embodiment, a subject with an underlying condition such as cardiovascular disease, diabetes, obesity, or atherosclerosis is treated with an herbal composition as described above to reduce the risk of damaging effects on the brain caused by neuroinflammation. In a specific embodiment, the underlying condition is diabetes. In a specific embodiment, a diabetic subject is treated with an herbal composition as described above to improve post-stroke recovery after ischemic stroke.

[0060] In one embodiment of any of the above aspects, the herbal composition is a pharmaceutical composition.The pharmaceutical composition may further comprise one or more pharmaceutically acceptable carriers or excipients.

[0061] In one embodiment of any of the above aspects, the herbal composition comprises an extract of the aforementioned herbal components.

[0062] In one embodiment of any one of the above aspects, the herbal composition comprises 5 times the weight / weight ratio of Astragalus root or its extract compared to the weight of each of the other herbal components or extracts thereof present.

[0063] In one embodiment of any of the above aspects, the pharmaceutical composition is MLC901.

[0064] In one embodiment of any of the above aspects, the pharmaceutical composition is MLC1501.

[0065] In one embodiment of any of the above aspects, the neuroinflammation is a result of insult, such as an injury, for example an injury selected from spinal cord injury, stroke, traumatic brain injury, subarachnoid hemorrhage, subdural hemorrhage, or epidural hemorrhage.

[0066] In a specific embodiment of any of the above aspects, the neuroinflammation occurs after an ischemic stroke. In this embodiment, treating a subject with the herbal composition described herein (including treating with MLC901 or treating with MLC1501) provides:

[0067] 1. Increase the rate of functional independence / autonomy recovery from about 3 months to about 24 months after the first administration of the herbal composition. In one aspect, the functional independence / autonomy recovery is due to controlling, regulating, protecting and / or strengthening the blood-brain barrier against the damaging effects of neuroinflammation;

[0068] 2. Reduce the time to achieve functional independence / autonomy by approximately 18 months. In one aspect, functional independence / autonomy is due to controlling, regulating, protecting and / or strengthening the blood-brain barrier to resist the damaging effects of neuroinflammation;

[0069] 3. Accelerate the achievement of functional independence / autonomy recovery. In one aspect, functional independence / autonomy recovery is due to controlling, regulating, protecting and / or strengthening the blood-brain barrier to resist the damaging effects of neuroinflammation;

[0070] 4. Reduced length of hospital stay. In one aspect, the reduction in length of hospital stay is the result of controlling, regulating, protecting and / or strengthening the blood-brain barrier to resist the damaging effects of neuroinflammation;

[0071] 5. Reduce the time and / or cost associated with functional independence / autonomy recovery. In one aspect, functional independence / autonomy recovery is due to controlling, regulating, protecting and / or strengthening the blood-brain barrier to resist the damaging effects of neuroinflammation;

[0072] and / or

[0073] 6. Improved quality of life. In one aspect, the improvement is the result of controlling, regulating, protecting and / or strengthening the blood-brain barrier to resist the damaging effects of neuroinflammation.

[0074] In a specific embodiment of any of the above aspects, neuroinflammation occurs after a trauma-induced neurodegenerative disease, such as a traumatic brain injury (TBI) or a spinal cord injury. In this embodiment, treatment of a subject with the herbal composition described herein, including treatment with MLC901 or treatment with MLC1501, provides a reduction in the severity of symptoms following injury. In a specific embodiment, the reduction in the severity of symptoms following injury is achieved by controlling, regulating, protecting and / or strengthening the blood-brain barrier to resist the damaging effects of neuroinflammation. After TBI, treatment of a subject with the herbal composition described herein, including treatment with MLC901 or treatment with MLC1501, may also provide improved recovery in cognitive areas, such as complex attention and / or executive function, and / or may have a positive impact on anxiety or depression symptoms and / or quality of life. In a specific embodiment, recovery in cognitive areas is achieved by controlling, regulating, protecting and / or strengthening the blood-brain barrier to resist the damaging effects of neuroinflammation.

[0075] In specific embodiments of any of the above aspects, treatment of a subject with the herbal composition described herein, including treatment with MLC901 or treatment with MLC1501, improves mood disorders associated with neurodegenerative diseases. In specific embodiments, improvement of mood disorders is achieved by controlling, regulating, protecting and / or strengthening the blood-brain barrier to resist the damaging effects of neuroinflammation.

[0076] In certain embodiments of any of the above aspects, the neuroinflammation is due to the subject having dementia or Alzheimer's disease. In this embodiment, treating the subject with the herbal composition described herein (including treatment with MLC901 or treatment with MLC1501) provides:

[0077] 1. Improved recovery of cognitive function (e.g., memory, practice, and / or language) within about 3 months to about 48 months after the first administration of the herbal composition. In one aspect, the improved recovery is due to control, regulation, protection, and / or

[0078] Or it may be caused by strengthening the blood-brain barrier to resist the damaging effects of neuroinflammation;

[0079] 2. Slowing disease progression. In one aspect, the slowing of disease progression is due to controlling, regulating, protecting and / or strengthening the blood-brain barrier to resist the damaging effects of neuroinflammation; and / or

[0080] 3. Disease Modifying Effects. In one aspect, the disease modifying effects are due to controlling, regulating, protecting and / or strengthening the blood-brain barrier against the damaging effects of neuroinflammation.

[0081] In one embodiment of any of the above aspects, neuroinflammation is the result of damage, such as infection, for example, infection of the brain or central nervous system (e.g., encephalitis or bacterial, viral or fungal meningitis), or severe infection such as sepsis or abscess. Examples of viral infections include varicella zoster virus (VZV) infection, herpes simplex virus (HSV-1 or HSV-2), SARS-CoV-2 virus or its variants, Epstein-Barr virus, hepatitis B virus, hepatitis C virus, human immunodeficiency virus type 1 (HIV-1), human papillomavirus (HPV), human T-lymphotropic virus type 1 (HTLV-1), and Kaposi's sarcoma herpesvirus (KSHV). Exemplary fungi that can cause neuroinflammation include Cryptococcus neoformans, Candida albicans, Histoplasma capsulatum, Coccidioides immitis, Paracoccidioides brasiliensis, Aspergillus species, 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 may also be associated with parasitic infections.

[0082] In specific embodiments, neuroinflammation is due to systemic inflammation from the subject's peripheral immune system caused by an infection such as COVID or "long COVID." In this embodiment, treatment of the subject with the herbal composition described herein, including treatment with MLC901 or treatment with MLC1501, provides protection against the progression of the neurodegenerative process by controlling, regulating, protecting and / or strengthening the blood-brain barrier against the damaging effects of neuroinflammation.

[0083] In one embodiment of any of the above aspects, the neuroinflammation is a result of exposure to a toxin.

[0084] In one embodiment of any of the above aspects, neuroinflammation is associated with a neurodegenerative disease such as Alzheimer's disease, Parkinson's disease, Huntington's disease, frontotemporal dementia, amyotrophic lateral sclerosis, multiple sclerosis, 10-12 primary tauopathy, synucleinopathies (i.e., Lewy body dementia and multiple system atrophy), polyglutamine diseases including spinocerebellar ataxia, prion diseases, traumatic brain injury, chronic traumatic encephalopathy, stroke, and spinal cord injury.

[0085] In one embodiment, neuroinflammation is associated with a neuropsychiatric disorder such as depression, stress, post-traumatic stress disorder, schizophrenia, autism spectrum disorder, or epilepsy.

[0086] In one embodiment of any of the above aspects, neuroinflammation is associated with aging (eg, "inflammaging").

[0087] Other conditions that can lead to neuroinflammation include postoperative cognitive dysfunction, ischemia, brain tumors, chronic pain, headache, migraine, trigeminal neuralgia, cardiovascular disease and related risk factors (e.g. due to obesity, hypertension, diabetes, hyperlipidemia, smoking, etc.), chronic peripheral inflammatory disorders (e.g. rheumatoid arthritis, inflammatory bowel disease, psoriasis, etc.), gut microbiota disturbances (i.e. dysbiosis), and herpetic keratitis.

[0088] In one embodiment, neuroinflammation is due to systemic inflammation from the subject's peripheral immune system caused by a disturbance (i.e., dysbiosis) in the intestinal microbiota. In this embodiment, treatment of the subject with the herbal composition described herein, including treatment with MLC901 or treatment with MLC1501, provides protection against the progression of the neurodegenerative process by controlling, regulating, protecting and / or strengthening the blood-brain barrier against the damaging effects of neuroinflammation.

[0089] In one embodiment of any of the above aspects, the neuroinflammation is caused by a chemical intervention to treat the underlying condition or disease. An example of such a chemical intervention is the use of TNF inhibitors to treat autoimmune diseases such as rheumatoid arthritis.

[0090] In one embodiment of any of the above aspects, the neuroinflammation is caused by a mechanical intervention to treat the underlying condition or disease.

[0091] In one embodiment of any of the above aspects, the neuroinflammation is caused by a surgical intervention to treat the underlying condition or disease. Examples of surgical interventions include carotid endarterectomy, carotid angioplasty with or without stents, vertebral angioplasty with or without stents, intracranial angioplasty with or without stents, aneurysm coil embolization, vascular malformation occlusion, intracranial fistula repair, thrombectomy, extracranial-intracranial bypass, open heart surgery, heart surgery requiring a bypass machine, coronary artery bypass, coronary angioplasty with or without stents, valve surgery / valvuloplasty, valvulotomy, patent foramen ovale closure, and cardiac great vessel surgery.

[0092] 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.

[0093] In one embodiment of any of the above aspects, a method is provided, comprising administering a second agent that treats the underlying condition or disease in addition to the herbal composition herein that treats neuroinflammation.

[0094] In one embodiment of any of the above aspects, the subject is diagnosed as having neuroinflammation, such as by a healthcare professional.

[0095] 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, such as the levels of one or more cytokines selected from IL-1β, IL-6, and TNF-α.

[0096] In one embodiment of any of the above aspects, the subject is tested for neuroinflammation by measuring the levels of inflammasomes including NLRP3 and / or iNOS.

[0097] In one aspect, a subject confirmed to have neuroinflammation following testing (e.g., by identifying the subject as exhibiting elevated levels of pro-inflammatory chemokines and / or cytokines, such as 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 such as NLRP3 and / or iNOS) is treated with an effective amount of an herbal composition comprising at least four herbal components: Polygala tenuifolia (root of Polygala), Astragalus membranaceus (root of Astragalus), Ligusticum chuanxiong rhizome, and Angelica dahurica (root of Angelica sinensis), or extracts thereof.

[0098] In a further aspect, a subject confirmed to have neuroinflammation following testing (e.g., by identifying the subject as exhibiting elevated levels of pro-inflammatory chemokines and / or cytokines, such as 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 such as NLRP3 and / or iNOS) is treated with an effective amount of an herbal composition comprising at least four herbal components: Polygala root (root of Polygala), Astragalus root (root of Astragalus), Ligusticum chuanxiong rhizome, and Angelica root (root of Angelica sinensis), or extracts thereof, and at least one other herbal component selected from the group consisting of Salvia miltiorrhiza root and rhizome (root of Salvia miltiorrhiza), Paeonia rubra root (root of Paeonia rubra), Carthamus tinctorius flower, Prunus persica (seed of peach), and Acorus tatarinowii rhizome (rhizome of Acorus calamus), or extracts thereof.

[0099] In another aspect, a subject confirmed to have neuroinflammation following testing (e.g., by identifying the subject as exhibiting elevated levels of pro-inflammatory chemokines and / or cytokines, such as 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 such as NLRP3 and / or iNOS) is treated with an effective amount of an herbal composition consisting essentially of at least four of the herbal components: Polygala tenuifolia (root of Polygala), Astragalus membranaceus (root of Astragalus), Ligusticum chuanxiong rhizome, and Angelica dahurica (root of Angelica sinensis), or extracts thereof.

[0100] In yet another aspect, a subject confirmed to have neuroinflammation following testing (e.g., by identifying the subject as exhibiting pro-inflammatory chemokines and / or cytokines, such as 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, such as NLRP3 and / or iNOS) is treated with an effective amount of an herbal composition consisting essentially of at least nine herbal components: Polygala root (Polygala tenuifolia root), Astragalus root (Astragalus root), Ligusticum chuanxiong rhizome, Angelica root (Angelica root), Salvia miltiorrhiza root and rhizome (Salvia miltiorrhiza root), Paeonia rubra root (Paeonia rubra root), Carthamus tinctorius flower, Prunus persica (Peach seed), and Acorus tatarinowii rhizome (Acorus calamus rhizome), or extracts thereof.

[0101] In one embodiment of any of the above aspects, the herbal composition comprises an extract of the aforementioned herbal components.

[0102] In one embodiment of any one of the above aspects, the herbal composition comprises 5 times the weight / weight ratio of Astragalus root or its extract compared to the weight of each of the other herbal components or extracts thereof present.

[0103] In one embodiment of any of the above aspects, when the subject is confirmed to suffer from neuroinflammation after testing, the herbal composition used to treat the subject is MLC901.

[0104] In one embodiment of any of the above aspects, when the subject is confirmed to suffer from neuroinflammation after testing, the herbal composition used to treat the subject is MLC1501.

[0105] In one aspect, a subject at risk for neuroinflammation due to insult (e.g., due to the aforementioned conditions, diseases, injuries, infection, aging, or chemical, mechanical or surgical intervention) is treated to prevent or limit neuroinflammation by administering (before, concurrently with, or after insult) an effective amount of an herbal composition comprising at least four herbal components: Polygala tenuifolia (root of Polygala), Astragalus membranaceus (root of Astragalus), Ligusticum chuanxiong rhizome, and Angelica dahurica (root of Angelica sinensis), or extracts thereof.

[0106] In a further aspect, a subject at risk for neuroinflammation due to insult (e.g., due to the aforementioned conditions, diseases, injury, infection, aging, or chemical, mechanical or surgical intervention) is treated to prevent or limit neuroinflammation by administering (before, simultaneously with, or after insult) an effective amount of a herbal composition comprising at least four herbal components: Polygala root (Polygala tenuifolia root), Astragalus root (Astragalus root), Ligusticum chuanxiong rhizome, and Angelica dahurica root (Angelica root), or extracts thereof, and at least one other herbal component selected from the group consisting of Salvia miltiorrhiza root and rhizome (Danshen root), Paeonia rubra root (Red Peony root), Carthamus tinctorius flower, Prunus persica (Peach seed), and Acorus calamus rhizome (Acorus calamus rhizome), or extracts thereof.

[0107] In another aspect, a subject at risk for neuroinflammation due to insult (e.g., due to the aforementioned conditions, diseases, injuries, infection, aging, or chemical, mechanical or surgical intervention) is treated to prevent or limit neuroinflammation by administering (before, concurrently with, or after insult) an effective amount of an herbal composition consisting essentially of at least four herbal components: Polygala tenuifolia (root of Polygala), Astragalus membranaceus (root of Astragalus), Ligusticum chuanxiong rhizome, and Angelica dahurica (root of Angelica sinensis), or extracts thereof.

[0108] In another aspect, a subject at risk for neuroinflammation due to insult (e.g., due to the aforementioned conditions, diseases, injuries, infection, aging, or chemical, mechanical or surgical intervention) is treated to prevent or limit neuroinflammation by administering (before, simultaneously with, or after the insult) an effective amount of an herbal composition consisting essentially of at least nine herbal components: Polygala root (Polygala tenuifolia root), Astragalus root (Astragalus root), Ligusticum chuanxiong rhizome, Angelica dahurica root (Angelica root), Salvia miltiorrhiza root and rhizome (Salvia miltiorrhiza root), Paeonia rubra root (Paeonia rubra root), Carthamus tinctorius flower, Prunus persica (Peach seed), and Acorus calamus rhizome (Acorus calamus rhizome), or extracts thereof.

[0109] In one embodiment of any of the above aspects, the herbal composition comprises an extract of the aforementioned herbal components.

[0110] In one embodiment of any one of the above aspects, the herbal composition comprises 5 times the weight / weight ratio of Astragalus root or its extract compared to the weight of each of the other herbal components or extracts thereof present.

[0111] In one embodiment of any of the above aspects, when the subject is at risk for neuroinflammation due to injury, the herbal composition for preventing or limiting neuroinflammation is MLC901.

[0112] In one embodiment of any of the above aspects, when the subject is at risk for neuroinflammation due to injury, the herbal composition for preventing or limiting neuroinflammation is MLC1501.

[0113] In one aspect, a subject at risk of developing a disease or condition due to neuroinflammation is treated with an effective anti-inflammatory amount of an herbal composition to prevent or reduce the development of the disease or condition, the herbal composition comprising at least four herbal components: Polygala root (Polygala tenuifolia root), Astragalus root (Astragalus root), Ligusticum chuanxiong rhizome, and Angelica dahurica root (Angelica root), or extracts thereof.

[0114] In another aspect, a subject at risk of developing a disease or condition due to neuroinflammation is treated with an effective anti-inflammatory amount of an herbal composition to prevent or reduce the development of the disease or condition, the herbal composition comprising at least four herbal components: Polygala root (Polygala root), Astragalus root (Astragalus root), Ligusticum chuanxiong rhizome and Angelica root (Angelica root), or extracts thereof, and at least one other herbal component selected from the group consisting of Salvia miltiorrhiza root and rhizome (Danshen root), Paeonia rubra root (Red Peony root), Carthamus tinctorius flower, Prunus persica (Peach seed) and Acorus tatarinowii rhizome (Acorus calamus rhizome), or extracts thereof.

[0115] In another aspect, a subject at risk of developing a disease or condition due to neuroinflammation is treated with an effective anti-inflammatory amount of an herbal composition consisting essentially of at least four herbal components: Polygala tenuifolia (root of Polygala), Astragalus membranaceus (root of Astragalus), Ligusticum chuanxiong rhizome, and Angelica dahurica (root of Angelica sinensis), or extracts thereof, to prevent or reduce the development of the disease or condition.

[0116] In yet another aspect, a subject at risk of developing a disease or condition due to neuroinflammation is treated with an effective anti-inflammatory amount of an herbal composition consisting essentially of at least nine herbal components: Polygala root (Polygala root), Astragalus root (Astragalus root), Ligusticum chuanxiong rhizome, Angelica root (Angelica root), Salvia miltiorrhiza root and rhizome (Salvia root), Paeonia rubra root (Red Peony root), Carthamus tinctorius flower, Prunus persica (Peach seed) and Acorus tatarinowii rhizome (Acorus calamus rhizome), or extracts thereof, to prevent or reduce the development of the disease or condition.

[0117] In one embodiment of any of the above aspects, the herbal composition comprises an extract of the aforementioned herbal components.

[0118] In one embodiment of any one of the above aspects, the herbal composition comprises 5 times the weight / weight ratio of Astragalus root or its extract compared to the weight of each of the other herbal components or extracts thereof present.

[0119] In one embodiment of any of the above aspects, when the subject is at risk of developing a disease or disorder due to neuroinflammation, the herbal composition for preventing or reducing the progression of the disease or disorder is MLC901.

[0120] In one embodiment of any of the above aspects, when the subject is at risk of developing a disease or disorder due to neuroinflammation, the herbal composition for preventing or reducing the progression of the disease or disorder is MLC1501.

[0121] The aforementioned "disease or condition" may be, for example, brain injury, brain swelling, blood-brain barrier leakage, cerebral stroke, transient ischemic attack or neuropsychiatric disorder (eg, depression, stress, post-traumatic stress disorder, schizophrenia, autism spectrum disorder or epilepsy).

[0122] NeuroAid II, referred to herein as "MLC901", is a composition containing extracts of nine herbal components (Astragalus root, Salvia root, Paeonia lactiflora root, Ligusticum chuanxiong rhizome, Angelica root, Carthamus tinctorius, Semen persica, Polygala root, and Acorus calamus rhizome). In MLC901, the Astragalus root extract is present at a weight / weight ratio of 5 times that of each of the other herbal component extracts present. It is currently marketed as an oral treatment to support recovery after stroke.

[0123] MLC1501 is a composition containing extracts of four herbal components (Astragalus root, Chuanxiong rhizome, Polygala root and Angelica root). In MLC1501, the Astragalus root extract is present at a weight / weight ratio of 5 times that of each of the other herbal component extracts present. Human clinical trials are currently underway to assess efficacy in post-stroke recovery.

[0124] MLC901 and MLC1501 and their preparation are described in published PCT application number WO 2017 / 048191A1, the contents of which are incorporated herein by reference in their entirety.

[0125] In one embodiment of any of the above aspects, the herbal composition of the present method consists essentially of at least four herbal components: Polygala root, Astragalus root, Ligusticum chuanxiong rhizome and Angelica root, or extracts thereof, and any 1, 2, 3, 4, or 5 of the following herbal components: Salvia miltiorrhiza root, Paeonia lactiflora root, Carthamus tinctorius, Semen persica and Acorus calamus rhizome, or extracts thereof. Such herbal compositions and their preparation are described in published PCT application numbers WO 2007 / 106049A1, WO 2010 / 053456A1, WO 2010 / 110755A1 and WO 2013 / 141818A1, the contents of which are incorporated herein by reference in their entirety.

[0126] The herbal compositions herein (including MLC901 and MLC1501) are generally extremely effective in treating or alleviating neuroinflammation in subjects who have already suffered from stroke (especially ischemic stroke), thereby contributing to post-stroke recovery. However, it is known that patients with underlying conditions (especially where the underlying conditions have inflammatory components, such as in diabetes) have a poor prognosis for functional recovery after stroke and standard therapy (including rehabilitation therapy). Stroke in diabetic patients can lead to increased mortality, as well as long-term neurological and functional disorders under standard care treatment. Nevertheless, it has been found that the herbal compositions herein (including MLC901 and MLC1501) are effective in contributing to post-stroke recovery in diabetes.

[0127] Therefore, in one aspect of the present disclosure, there is provided a method for treating or reducing neuroinflammation in a diabetic subject after ischemic stroke, the method comprising administering to the subject an effective amount of an herbal composition comprising at least four herbal components: Polygala tenuifolia, Astragalus membranaceus, Ligusticum chuanxiong and Angelica sinensis, their roots or rhizomes, or extracts thereof.

[0128] In one embodiment of the above aspects, the herbal composition comprising four herbal components: Polygala tenuifolia, Astragalus membranaceus, Ligusticum chuanxiong and Angelica sinensis, their roots or rhizomes, or extracts thereof further comprises at least one herbal component selected from the group consisting of Salviae Miltiorrhizae (red sage), Paeoniae Rubra (red peony), flower of Carthamus Tinctorius (safflower), Semen Persicae (peach seed) and Acori Tatarinowii (grassleaf sweetflag), their roots or rhizomes, or extracts thereof.

[0129] In another aspect, a method for treating or reducing neuroinflammation in a diabetic subject after ischemic stroke is provided, the method comprising administering to the subject an effective amount of an herbal composition consisting essentially of at least four herbal components: Polygala tenuifolia, Astragalus membranaceus, Ligusticum chuanxiong, Angelica sinensis, their roots or rhizomes, or extracts thereof.

[0130] In another aspect, a method for treating or reducing neuroinflammation in a diabetic subject after ischemic stroke is provided, the method comprising administering to the subject an effective amount of an herbal composition consisting essentially of nine herbal components: Polygala tenuifolia, Astragalus membranaceus, Ligusticum chuanxiong, Angelica sinensis, Salvia miltiorrhiza, Paeonia lactiflora, Carthamus tinctorius flower, Persica persica (peach seed), and Acorus tatarinowii, their roots or rhizomes, or extracts thereof.

[0131] In one embodiment of any of the above aspects, the herbal composition is a pharmaceutical composition. The pharmaceutical composition may further comprise 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.

[0132] In certain aspects of the present disclosure, a method of treating or reducing neuroinflammation in a diabetic subject following ischemic stroke is provided, the method comprising administering an effective amount of MLC901 to the subject.

[0133] In certain aspects of the present disclosure, there is provided a method of treating or reducing neuroinflammation in a diabetic subject after ischemic stroke, the method comprising administering an effective amount of MLC1501 to the subject.

[0134] In a particular aspect of the present disclosure, a method for treating a diabetic subject after ischemic stroke is provided, the method comprising administering an effective amount of MLC901 to the subject to improve post-stroke recovery. In one embodiment, the subject is receiving standard care after stroke, such as treatment with an antiplatelet agent (e.g., aspirin) and / or rehabilitation therapy. In one embodiment, compared with a diabetic subject who only receives standard care after stroke, a diabetic subject who experiences standard care after stroke and co-administered MLC901 shows an excellent recovery rate. In one embodiment, excellence is demonstrated within about 3, 6, 9, 12, 18, or 24 months after the occurrence of a stroke.

[0135] In a particular aspect of the present disclosure, a method for treating a diabetic subject after ischemic stroke is provided, the method comprising administering an effective amount of MLC1501 to the subject to improve post-stroke recovery. In one embodiment, the subject is receiving standard care after stroke, such as treatment with an antiplatelet agent (e.g., aspirin) and / or rehabilitation therapy. In one embodiment, compared with diabetic subjects who only receive standard care after stroke, diabetic subjects who experience standard care after stroke and co-administered MLC1501 show excellent recovery rates. In one embodiment, excellence is demonstrated within about 3, 6, 9, 12, 18, or 24 months after stroke occurs.

[0136] The pharmaceutical composition herein may optionally include one or more pharmaceutically acceptable additives, carriers and / or diluents. The example of a pharmaceutically acceptable additive includes a pharmaceutically acceptable excipient, a buffer, an adjuvant, a stabilizer, a diluent, a filler, a preservative, a lubricant or other pharmaceutically acceptable materials known to those skilled in the art or as described herein. Examples of suitable pharmaceutical carriers or diluents include phosphate buffered saline solutions, water, emulsions (e.g., oil / water emulsions), various types of wetting agents, sterile solutions, etc. The example of an excipient that can be used includes, for example, sugar, starch, cellulose, gum, protein, dextrin and maltodextrin. Various preparations are well known and discussed in depth in the latest edition of Remington's Pharmaceutical Sciences (Maack Publishing, Easton PA). In at least some embodiments, the pharmaceutical composition described herein includes an excipient, such as dextrin or maltodextrin.

[0137] In one embodiment of any of the above aspects, the composition (e.g., pharmaceutical composition) of the present disclosure may be included in a kit. In addition to the herbal components, the kit may also include instructions for use. The kit may be promoted, distributed, and / or sold as a unit for performing one of the aspects of the present disclosure.

[0138] Generally speaking, the pharmaceutical compositions of the present disclosure can be prepared according to methods known to those of ordinary skill in the art.

[0139] The composition may be, for example, a solution, a suspension, a liquid, chopped herbs, a powder, a paste, aqueous, non-aqueous, or any combination thereof.

[0140] The compositions of the present disclosure (e.g., pharmaceutical compositions) can be administered in liquid, semi-liquid or solid form by any suitable route, such as oral, parenteral, intravenous, subcutaneous, intradermal, intraperitoneal or topical, and formulated in a manner suitable for each route of administration. The term "administering" and variants of the term (including "administering" and "administration") include contacting, administering, delivering or providing the compositions of the present disclosure to an organism or surface by any appropriate means.

[0141] The herbal components may be administered in a therapeutically effective amount (as a single dose or as part of a series of doses). An "effective amount" or "therapeutically effective amount" refers to an amount administered to achieve a physiologically significant effect. An agent is physiologically significant if the agent is present in an amount that produces a detectable change in the physiology of the recipient patient, thereby achieving a beneficial or desired result.

[0142] The exact amount required will vary from subject to subject, depending on factors such as the species being treated, the age, weight and general health of the subject, the condition being treated and the severity of the condition, the mode of administration, the sex of the subject, diet, time and frequency of administration, drug combination, and tolerance / response to therapy.

[0143] 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 once or more (e.g., 1, 2, 3, or 4 times) per day. In a specific embodiment, MLC901 is administered in the form of 2 capsules, taken 3 times per day. In a specific embodiment, MLC1501 is administered in the form of 4 capsules, taken 2 times per day. For patients with dysphagia, the capsule can be opened and the powder diluted in water, which can be drunk directly or injected via a gastric tube.

[0144] In one embodiment of any of the above aspects, the daily dose for a patient is about 500 mg to about 8 g, such as 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 or multiple unit doses (tablets, capsules, etc.) taken on a given day. However, it should be understood that the dose can vary according to the patient's requirements and the severity of the condition being treated, etc.

[0145] The duration of treatment is several days, or more typically several months (e.g., three months), which can be adjusted according to the patient's condition. In one embodiment of any of the above aspects, the treatment lasts for about 12 weeks. In another embodiment of any of the above aspects, the treatment lasts for about 24 weeks. In another embodiment of any of the above aspects, the treatment lasts for about 36 weeks. In another embodiment of any of the above aspects, the treatment lasts for about 48 weeks. In another embodiment of any of the above aspects, the treatment lasts for longer than about 48 weeks.

[0146] 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, such as caused by injury, infection, exposure to toxins, neurodegenerative diseases, aging (e.g., "inflammatory aging"), or by chemical, mechanical, or surgical intervention to treat an underlying condition or disease 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., 1 to 5 days, 1 to 4 days, 1 to 3 days, or 1 to 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, such as within 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 hours after the injury.

[0147] In one embodiment of any of the above aspects, the treatment regimen is designed for acute treatment.

[0148] In one embodiment of any of the above aspects, the treatment regimen is designed for long-term treatment.

[0149] In one embodiment of any of the above aspects, the treatment regimen is designed for post-acute treatment.

[0150] In one embodiment of any of the above aspects, the herbal composition of the present disclosure (e.g., MLC901 or MLC1501) can be administered to a subject in combination with an active agent that treats an underlying condition or disease 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 can be administered at the same time (e.g., simultaneously) or at different times (e.g., sequentially) as the herbal composition of the present disclosure and in different time periods that may be separate or overlapping from each other.

[0151] The one or more additional active agents may be administered by the same or different routes as the herbal composition of the present disclosure. The one or more additional active agents used and the appropriate routes of administration and dosage levels are known to or can be readily determined by those skilled in the art. Generally, as is well known in the medical field, the dosage regimen may depend on a variety of factors, including the patient's size, body surface area, age, the specific compound to be administered, sex, time and route of administration, general health, and other drugs administered simultaneously. Although individual needs vary, it is within the skill of the art to determine the optimal range of effective amounts of each component. Generally, the dosage of the one or more additional active agents will be the same or similar to the dosage administered when the active agent is not used with 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 comprising the herbal composition of the present disclosure, or the one or more additional active agents may be provided separately (i.e., not as part of a composition or kit providing the herbal composition of the present disclosure).

[0152] In addition to the above embodiments, the present disclosure also includes the following specific embodiments:

[0153] Embodiment 1: A herbal composition for treating neuroinflammation in a subject, the herbal composition comprising at least four herbal components: Polygala tenuifolia (root of Polygala), Astragalus membranaceus (root of Astragalus), Ligusticum chuanxiong rhizome and Danggui root (root of Angelica sinensis), or extracts thereof.

[0154] Embodiment 2: The herbal composition for use according to embodiment 1, wherein the herbal composition further comprises at least one herbal component selected from the group consisting of Salvia miltiorrhiza root and rhizome (root of Salvia miltiorrhiza), Paeonia rubra root (root of Paeonia rubra), Carthamus tinctorius flower, Prunus persica (peach seed) and Acorus calamus rhizome (rhizome of Acorus calamus), or extracts thereof.

[0155] Embodiment 3: The herbal composition for use according to Example 1, wherein the herbal composition consists essentially of at least four herbal components: Polygala root (Polygala tenuifolia root), Astragalus root (Astragalus root), Ligusticum chuanxiong rhizome and Angelica acutiloba root (Angelica root), or extracts thereof.

[0156] Embodiment 4: The herbal composition for use according to Example 1, wherein the herbal composition consists essentially of at least nine herbal components: Polygala root (Polygala root), Astragalus root (Astragalus root), Ligusticum chuanxiong rhizome, Angelica root (Angelica root), Salvia miltiorrhiza root and rhizome (Salvia root), Paeonia rubra root (Paeonia rubra root), Carthamus tinctorius flower, Prunus persica (Peach seed) and Acorus calamus rhizome (Acorus calamus rhizome), or extracts thereof.

[0157] Embodiment 5: The herbal composition for use according to any one of embodiments 1 to 4, wherein the herbal composition is a pharmaceutical composition.

[0158] Embodiment 6: The herbal composition for use according to embodiment 5, wherein the pharmaceutical composition further comprises a pharmaceutically acceptable carrier or excipient.

[0159] Embodiment 7: The herbal composition for use according to Example 6, wherein the pharmaceutical composition is composition MLC901. Embodiment 8: The herbal composition for use according to Example 6, wherein the pharmaceutical composition is composition MLC1501. 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.

[0160] 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.

[0161] Embodiment 11: The herbal composition for use according to any one of embodiments 1 to 10, wherein the neuroinflammation is a result of injury.

[0162] 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.

[0163] Embodiment 13: The herbal composition for use according to any one of embodiments 1 to 10, wherein the neuroinflammation is a result of infection.

[0164] Embodiment 14: The herbal composition for use according to embodiment 13, wherein the infection is an infection of the brain or central nervous system (such as encephalitis or bacterial, viral or fungal meningitis).

[0165] Embodiment 15:The herbal composition for use according to Example 13, wherein the infection is a viral infection [e.g., varicella zoster virus (VZV) infection, herpes simplex virus (HSV-1 or HSV-2), SARS-CoV-2 virus or variants thereof, Epstein-Barr virus, hepatitis B virus, hepatitis C virus, human immunodeficiency virus type 1 (HIV-1), human papillomavirus (HPV), human T-lymphotropic virus type 1 (HTLV-1), or Kaposi's sarcoma herpesvirus (KSHV)], a fungal infection [e.g., wherein the fungus is selected from the group consisting of Cryptococcus neoformans, Candida albicans, Histoplasma capsulatum, Coccidioides immitis, Paracoccidioides brasiliensis, Aspergillus species, and Zygomycetes], a bacterial infection [e.g., wherein the bacteria is selected from the group consisting of Listeria monocytogenes, Borrelia burgdorferi, Neisseria meningitidis, Streptococcus pneumoniae, Staphylococcus aureus, Haemophilus influenzae, Mycobacterium, and Brucella species], or a parasitic infection.

[0166] 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.

[0167] Embodiment 17: The herbal composition for use according to any one of embodiments 1 to 10, 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, 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].

[0168] Embodiment 18: The herbal composition for use according to any one of embodiments 1 to 10, wherein the neuroinflammation is associated with a neuropsychiatric disorder (e.g., depression, stress, post-traumatic stress disorder, schizophrenia, autism spectrum disorder or epilepsy).

[0169] Embodiment 19: The herbal composition for use according to any one of embodiments 1 to 10, wherein the neuroinflammation is associated with aging (eg, "inflammaging").

[0170] 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 condition selected from the group consisting of: postoperative cognitive dysfunction, ischemia, brain tumors, chronic pain, headache, migraine, trigeminal neuralgia, cardiovascular disease and related risk factors (e.g. due to obesity, hypertension, diabetes, hyperlipidemia, smoking, etc.), chronic peripheral inflammatory disorders (e.g. rheumatoid arthritis, inflammatory bowel disease, psoriasis, etc.), gut microbiota disturbances (i.e. dysbiosis) and herpetic keratitis.

[0171] Embodiment 21: The herbal composition for use according to any one of embodiments 1 to 10, wherein the neuroinflammation is caused by a chemical intervention to treat the underlying condition or disease. An example of such a chemical intervention is the use of TNF inhibitors to treat autoimmune diseases such as rheumatoid arthritis.

[0172] Embodiment 22: 14. The herbal composition for use according to any one of embodiments 1 to 10, wherein the neuroinflammation is caused by a mechanical intervention to treat the underlying condition or disease.

[0173] Embodiment 23: 14. The herbal composition for use according to any one of embodiments 1 to 10, wherein the neuroinflammation is caused by surgical intervention to treat the underlying condition or disease.

[0174] Embodiment 24: The herbal composition for use according to Example 23, wherein the surgical intervention is selected from the group consisting of carotid endarterectomy, carotid angioplasty with or without stent, vertebral angioplasty with or without stent, intracranial angioplasty with or without stent, aneurysm coil embolization, vascular malformation occlusion, intracranial fistula repair, thrombectomy, extracranial-intracranial bypass, open heart surgery, heart surgery requiring a bypass machine, coronary artery bypass grafting, coronary angioplasty with or without stent, valve surgery / valvuloplasty, valvulotomy, patent foramen ovale closure and cardiac great vessel surgery.

[0175] Embodiment 25: The herbal composition for use according to any one of embodiments 21 to 24, wherein said herbal composition is administered prior to, simultaneously with or after administration of said intervention to treat the underlying condition or disease.

[0176] Embodiment 26: 26. The herbal composition for use according to any one of embodiments 1 to 25, wherein the subject is tested for neuroinflammation and confirmed to have neuroinflammation prior to administering the herbal composition.

[0177] Embodiment 27:The herbal composition for use according to embodiment 26, wherein said testing shows that said subject exhibits elevated levels of pro-inflammatory chemokines and / or cytokines.

[0178] Embodiment 28: The herbal composition for use according to embodiment 26, wherein said testing shows that said 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.

[0179] Embodiment 29: An herbal composition for preventing or limiting neuroinflammation in a subject at risk of neuroinflammation due to injury, the herbal composition comprising at least four herbal components: Polygala tenuifolia (root of Polygala tenuifolia), Astragalus membranaceus (root of Astragalus membranaceus), Ligusticum chuanxiong rhizome and Angelica acutiloba (root of Angelica sinensis), or extracts thereof.

[0180] Embodiment 30: The herbal composition for use according to embodiment 29, wherein the herbal composition further comprises at least one herbal component selected from the group consisting of Salvia miltiorrhiza root and rhizome (root of Salvia miltiorrhiza), Paeonia rubra root (root of Paeonia rubra), Carthamus tinctorius flower, Prunus persica (peach seed) and Acorus calamus rhizome (rhizome of Acorus calamus), or extracts thereof.

[0181] Embodiment 31: The herbal composition for use according to Example 29, wherein the herbal composition consists essentially of at least four herbal components: Polygala tenuifolia (root of Polygala tenuifolia), Astragalus membranaceus (root of Astragalus membranaceus), Ligusticum chuanxiong rhizome and Danggui root (root of Angelica sinensis), or extracts thereof.

[0182] Embodiment 32: An herbal composition for use according to Example 29, wherein the herbal composition consists essentially of at least nine herbal components: Polygala root (Polygala root), Astragalus root (Astragalus root), Ligusticum chuanxiong rhizome, Angelica root (Angelica root), Salvia miltiorrhiza root and rhizome (Salvia root), Paeonia rubra root (Paeonia rubra root), Carthamus tinctorius flower, Prunus persica (Peach seed) and Acorus calamus rhizome (Acorus calamus rhizome), or extracts thereof.

[0183] Embodiment 33: The herbal composition for use according to any one of embodiments 29 to 32, wherein the herbal composition is a pharmaceutical composition.

[0184] Embodiment 34: The herbal composition for use according to embodiment 33, wherein the pharmaceutical composition further comprises a pharmaceutically acceptable carrier or excipient.

[0185] Embodiment 35:The herbal composition for use according to embodiment 34, wherein the pharmaceutical composition is composition MLC901.

[0186] Embodiment 36: The herbal composition for use according to embodiment 34, wherein the pharmaceutical composition is composition MLC1501.

[0187] 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.

[0188] 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.

[0189] Embodiment 39: The herbal composition for use according to any one of embodiments 29 to 38, wherein the damage is caused by a chemical, mechanical or surgical intervention as described in any one of embodiments 21 to 24 to treat the underlying condition or disease.

[0190] Embodiment 40: A herbal composition for treating a subject at risk of developing a disease or condition due to neuroinflammation to prevent or alleviate the development of the disease or condition, the herbal composition comprising at least four herbal components: Polygala tenuifolia (root of Polygala), Astragalus membranaceus (root of Astragalus), Ligusticum chuanxiong rhizome and Angelica acutiloba (root of Angelica sinensis), or extracts thereof.

[0191] Embodiment 41: The herbal composition for use according to embodiment 40, wherein the herbal composition further comprises at least one herbal component selected from the group consisting of Salvia miltiorrhiza root and rhizome (root of Salvia miltiorrhiza), Paeonia rubra root (root of Paeonia rubra), Carthamus tinctorius flower, Prunus persica (peach seed) and Acorus calamus rhizome (rhizome of Acorus calamus), or extracts thereof.

[0192] Embodiment 42: The herbal composition for use according to embodiment 40, wherein the herbal composition consists essentially of at least four herbal components: Polygala tenuifolia (root of Polygala tenuifolia), Astragalus membranaceus (root of Astragalus membranaceus), Ligusticum chuanxiong rhizome and Danggui root (root of Angelica sinensis), or extracts thereof.

[0193] Embodiment 43: An herbal composition for use according to Example 40, wherein the herbal composition consists essentially of at least nine herbal components: Polygala root (Polygala root), Astragalus root (Astragalus root), Ligusticum chuanxiong rhizome, Angelica root (Angelica root), Salvia miltiorrhiza root and rhizome (Salvia root), Paeonia rubra root (Paeonia rubra root), Carthamus tinctorius flower, Prunus persica (Peach seed) and Acorus calamus rhizome (Acorus calamus rhizome), or extracts thereof.

[0194] Embodiment 44: The herbal composition for use according to any one of embodiments 40 to 43, wherein the herbal composition is a pharmaceutical composition.

[0195] Embodiment 45: The herbal composition for use according to embodiment 44, wherein the pharmaceutical composition further comprises a pharmaceutically acceptable carrier or excipient.

[0196] Embodiment 46: The herbal composition for use according to embodiment 45, wherein the pharmaceutical composition is composition MLC901.

[0197] Embodiment 47: The herbal composition for use according to embodiment 45, wherein the pharmaceutical composition is composition MLC1501.

[0198] 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.

[0199] Embodiment 49: The herbal composition for use according to any one of embodiments 1 to 48, wherein the subject suffers from brain damage.

[0200] Embodiment 50: The herbal composition for use according to any one of embodiments 1 to 48, wherein the subject suffers from brain swelling.

[0201] Embodiment 51: The herbal composition for use according to any one of embodiments 1 to 48, wherein the subject suffers from a leaky blood-brain barrier.

[0202] Embodiment 52: The herbal composition for use according to any one of embodiments 1 to 48, wherein the subject suffers from a stroke.

[0203] Embodiment 53: The herbal composition for use according to any one of embodiments 1 to 48, wherein the subject suffers from a neuropsychiatric disorder (e.g., depression, stress, post-traumatic stress disorder, schizophrenia, autism spectrum disorder or epilepsy).

[0204] Embodiment 54: A herbal composition for treating neuroinflammation in a subject suffering from diabetes, the herbal composition comprising Polygala tenuifolia (root of Polygala tenuifolia), Astragalus membranaceus (root of Astragalus membranaceus), Ligusticum chuanxiong rhizome and Danggui root (root of Angelica sinensis), or extracts thereof.

[0205] Embodiment 55:A herbal composition for treating neuroinflammation in a subject with diabetes, the herbal composition comprising Polygala root (root of Polygala), Astragalus root (root of Astragalus), Ligusticum chuanxiong rhizome, Angelica root (root of Angelica sinensis), Salvia miltiorrhiza root and rhizome (root of Salvia miltiorrhiza), Paeonia rubra root (root of Paeonia rubra), Carthamus tinctorius flower, Prunus persica (seed of peach) and Acorus calamus rhizome (rhizome of Acorus calamus), or extracts thereof.

[0206] Embodiment 56: The herbal composition for use according to Example 54, wherein the herbal composition consists essentially of Polygala tenuifolia (root of Polygala tenuifolia), Astragalus membranaceus (root of Astragalus membranaceus), Ligusticum chuanxiong rhizome and Danggui root (root of Angelica sinensis), or extracts thereof.

[0207] Embodiment 57: An herbal composition for use according to Example 55, wherein the herbal composition consists essentially of Polygala root (Polygala root), Astragalus root (Astragalus root), Ligusticum chuanxiong rhizome, Angelica root (Angelica root), Salvia miltiorrhiza root and rhizome (Salvia root), Paeonia rubra root (Paeonia rubra root), Carthamus tinctorius flower, Prunus persica (Peach seed) and Acorus calamus rhizome (Acorus calamus rhizome), or extracts thereof.

[0208] Embodiment 58: The herbal composition for use according to any one of embodiments 54 to 57, wherein the herbal composition is a pharmaceutical composition.

[0209] Embodiment 59: The herbal composition for use according to embodiment 58, wherein the pharmaceutical composition further comprises a pharmaceutically acceptable carrier or excipient.

[0210] Embodiment 60: The herbal composition for use according to embodiment 58, wherein the pharmaceutical composition is composition MLC901.

[0211] Example 61: The herbal composition for use according to embodiment 58, wherein the pharmaceutical composition is composition MLC1501.

[0212] 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.

[0213] 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.

[0214] Embodiment 64:The herbal composition for use according to any one of embodiments 54 to 63, wherein the subject is tested for neuroinflammation and confirmed to have neuroinflammation prior to administering the herbal composition.

[0215] Embodiment 65: The herbal composition for use according to embodiment 64, wherein said testing shows that said subject exhibits elevated levels of pro-inflammatory chemokines and / or cytokines.

[0216] Embodiment 66: The herbal composition for use according to embodiment 64, wherein said testing shows that said 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.

[0217] Embodiment 67: The herbal composition for use according to any one of embodiments 54 to 66, wherein the neuroinflammation is caused by brain injury.

[0218] Embodiment 68: The herbal composition for use according to any one of embodiments 54 to 66, wherein the neuroinflammation is caused by brain swelling.

[0219] Embodiment 69: The herbal composition for use according to any one of embodiments 54 to 66, wherein the neuroinflammation is caused by blood-brain barrier leakage.

[0220] Embodiment 70: The herbal composition for use according to any one of embodiments 54 to 66, wherein the neuroinflammation is caused by cerebral stroke.

[0221] 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).

[0222] The examples below describe a study investigating the effects of MLC1501 on inflammation, and more specifically, its effects on the expression of various cellular markers involved in neuroinflammation. The study was divided into 2 parts:

[0223] 1 / Characterize the effect of MLC1501 after LPS-induced inflammation, focusing on the markers IL-1β, IL-6, TNF-α,

[0224] In vitro expression of NLRP3 and iNOS.

[0225] 2 / Characterize the effects of MLC1501 in a focal cerebral ischemia model, focusing on the in vivo expression of the pro-inflammatory cytokines IL-1β and IL-6.

[0226] The results obtained below for MLC1501 can also be achieved with other herbal compositions of the present disclosure, including MLC901.

[0227] Example 1

[0228] 1) Materials and methods

[0229] a) Cell culture

[0230] The BV-2 microglial cell line is derived from neonatal mouse microglia immortalized with v-raf / v-myc oncogenes (Blasi et al., 1990). It is cultured in DMEM "high glucose" medium supplemented with 5% fetal calf serum (FCS) and 1% penicillin / streptomycin (Gibco, Thermofisher Scientific, USA). The cells are maintained in a humidified incubator at 37°C and in an atmosphere containing 5% CO2. Once the cells reach 80% confluence, they are used for experiments.

[0231] b) In vitro treatment

[0232] Cells were pretreated with MLC1501 at a concentration of 250 μg / mL for 24 hours. For dose-response analysis, eight different concentrations of MLC1501 were selected: 0, 10, 50, 100, 250, 500, 1000, and 2000 μg / mL. c) In vitro inflammation model: LPS stimulation

[0233] In the absence of SVF but in the presence of MLC1501 (250 μg / mL), cells were stimulated with 100 ng / mL LPS (E. coli; O111:B4, Sigma-Aldrich) for 6 hours. The experimental scheme is shown in FIG1 .

[0234] d) Animals

[0235] Male C57Bl / 6JRj mice aged 7 weeks (at the time of receipt) were housed 5 mice per cage in an animal room maintained at an ambient temperature of 21±1° C. The animals had free access to food and water.

[0236] e) In vivo processing

[0237] A single 40 μg / kg dose of MLC1501 diluted in saline (vehicle) was injected intraperitoneally 60 minutes before the onset of ischemia (100 μL bolus). A schematic diagram of the protocol is shown in FIG2 .

[0238] f) MCAO model of ischemic stroke

[0239] The mice treated with vehicle and MLC1501 were subjected to a 60-minute focal ischemia model. The model consists of the following items: occlusion of the right middle cerebral artery (MCAO) for 60 minutes using a 6-0 filament (Doccol, Redlands, CA, USA) coated with silicone, followed by reperfusion (or not reperfusion in some cases). Cerebral blood flow in the target area was monitored by laser Doppler (Perimed, Craponne France) to control the severity of the pattern and the presence of reperfusion. Animals with less than 70% reduction in cerebral blood flow were excluded from the study (<1%). A group of control animals were subjected to a sham operation (filaments were inserted into the carotid artery without causing MCA occlusion). The animals were then placed in a thermostatically controlled recovery chamber (30°C to 31°C) and supplied with synthetic air for 30 hours.

[0240] g) mRNA extraction and quantitative rt-PCR

[0241] Microglia and ipsilateral brain hemisphere were collected 30 hours after each in vitro and in vivo protocol was started. mRNA from microglia and brain tissue was extracted with RNAzol (Euromedex, France), and then reverse transcribed using Superscript IV (Thermofisher Scientific, USA). The expression of mRNA encoding IL-1β, IL-6 and TNF-α genes was then determined by qPCR (SybrGreen 480Roche) using specific primers (Eurogentec, Angers, France) (sequences are shown in Figure 3). The results were normalized by the expression of housekeeping genes GAPDH or RPL0. Values ​​are expressed as 2-ΔΔCT normalized by control. Each sample was repeated.

[0242] h) Statistics and regression

[0243] Data are presented as mean ± SEM, and each sample is the average of 2 replicates. Regression was performed on GraphPad Prism software. Outliers were determined by the ROUT method (Q = 1%). Statistical analysis between different groups of animals was calculated using the Mann & Whitney test. The significance level was set at P < 0.05.

[0244] 2) Results

[0245] a) MLC1501 reduces the relative expression of genes involved in pro-inflammatory responses in an in vitro inflammation model:

[0246] 1 / Characterization of the anti-inflammatory effect of MLC1501 at a concentration of 250 μg / mL on the BV2 microglial cell line after LPS stimulation

[0247] We observed that 6 hours of LPS stimulation induced on cells not treated with MLC1501 significantly increased the mRNA expression levels of proinflammatory cytokine genes IL-1β, IL-6, and TNF-α. The same LPS stimulation also induced an increase in NLRP3 and iNOS gene expression in untreated cells (Figure 4). Compared with vehicle administration, the administration of MLC1501 at a concentration of 250 μg / mL significantly reduced the relative expression of IL-1β, IL-6, TNF-α, NLRP3, and iNOS genes after 6 hours of LPS stimulation (Figure 4).

[0248] 2 / Dose-response analysis of MLC1501 on the expression of three inflammatory markers: IL-1β, IL-6 and TNF-α

[0249] In order to perform dose response analysis, 8 different concentrations of MLC1501 were selected: 0, 10, 50, 100, 250, 500, 1000 and 2000 μg / mL. The administration of MLC1501 resulted in a comprehensive reduction in the relative overexpression of IL-1β, IL-6 and TNF-α in the presence of LPS in a dose-dependent manner. According to the regression performed, the IC50 (half-maximum inhibitory concentration) values ​​of 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).

[0250] (b) MLC1501 reduces the relative expression of pro-inflammatory genes in an in vivo focal ischemia model:

[0251] Focal ischemia induced by MCAO in the vehicle group induced high expression of proinflammatory genes. Administration of MLC1501 60 minutes before the onset of ischemia significantly reduced the relative expression of genes encoding cytokines IL-1β and IL-6 ( FIG. 6 ).

[0252] 3) Conclusion

[0253] The study showed that administration of MLC1501 at a concentration of 250 μg / mL reduced 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 in vitro after LPS stimulation and in vivo after focal cerebral ischemia.

[0254] 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 and patent application herein is not and should not be taken as an admission 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. A method for treating neuroinflammation in a subject, the method comprising administering to the subject an effective amount of an herbal composition comprising Radix Polygalae (root of Polygala), Radix Astragali (root of Astragalus), Rhizome Ligusticum Chuanxiong, and Radix Angelica sinensis (root of Angelica), or extracts thereof.

2. A method for treating neuroinflammation in a subject, the method comprising administering to the subject an effective amount of an herbal composition comprising Polygala root (root of Polygala), Astragalus root (root of Astragalus), Ligusticum chuanxiong rhizome, Angelica root (root of Angelica sinensis), Radix et Rhizome Salviae Miltiorrhizae (root of Salvia miltiorrhiza), Radix Paeoniae Rubra (root of Red Peony), flower of Carthamus Tinctorius (safflower), Semen Persicae (peach seeds), and Rhizome Acori Tatarinowii (rhizome of Acorus tatarinowii), or extracts thereof.

3. A method of treating neuroinflammation in a subject, the method comprising administering to the subject an effective amount of an herbal composition consisting essentially of: Polygala root (Polygala tenuifolia root), Astragalus root (Astragalus root), Ligusticum chuanxiong rhizome and Angelica dahurica root (Angelica root), or extracts thereof.

4. A method of treating neuroinflammation in a subject, the method comprising administering to the subject an effective amount of an herbal composition consisting essentially of: Polygala root (Polygala root), Astragalus root (Astragalus root), Ligusticum chuanxiong rhizome, Angelica root (Angelica root), Salvia miltiorrhiza root and rhizome (Salvia root), Paeonia rubra root (Paeonia rubra root), Carthamus tinctorius flower, Prunus persica (Peach seed) and Acorus calamus rhizome (Acorus calamus rhizome), or extracts thereof.

5. A method for treating neuroinflammation in a subject suffering from diabetes, the method comprising administering to the subject an effective amount of an herbal composition comprising Polygala tenuifolia (root of Polygala), Astragalus membranaceus (root of Astragalus), Ligusticum chuanxiong rhizome and Angelica acutiloba (root of Angelica sinensis), or extracts thereof.

6. A method for treating neuroinflammation in a subject suffering from diabetes, the method comprising administering to the subject an effective amount of an herbal composition comprising Polygala root (root of Polygala), Astragalus root (root of Astragalus), Ligusticum chuanxiong rhizome, Angelica root (root of Angelica sinensis), Salvia miltiorrhiza root and rhizome (root of Salvia miltiorrhiza), Paeonia rubra root (root of Paeonia rubra), Carthamus tinctorius flower, Prunus persica (peach seed) and Acorus calamus rhizome (rhizome of Acorus calamus), or extracts thereof.

7. A method of treating neuroinflammation in a subject suffering from diabetes, the method comprising administering to the subject an effective amount of an herbal composition consisting essentially of: Polygala root (Polygala tenuifolia root), Astragalus root (Astragalus root), Ligusticum chuanxiong rhizome and Angelica dahurica root (Angelica root), or extracts thereof.

8. A method of treating neuroinflammation in a subject suffering from diabetes, the method comprising administering to the subject an effective amount of an herbal composition consisting essentially of: Polygala root (Polygala root), Astragalus root (Astragalus root), Ligusticum chuanxiong rhizome, Angelica root (Angelica root), Salvia miltiorrhiza root and rhizome (Salvia root), Paeonia rubra root (Paeonia rubra root), Carthamus tinctorius flower, Prunus persica (Peach seed) and Acorus calamus rhizome (Acorus calamus rhizome), or extracts thereof.

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

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

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

13. The method of claim 11, wherein the infection is a viral infection [e.g., varicella zoster virus (VZV) infection, herpes simplex virus (HSV-1 or HSV-2), SARS-CoV-2 virus or variants thereof, Epstein-Barr virus, hepatitis B virus, hepatitis C virus, human immunodeficiency virus type 1 (HIV-1), human papillomavirus (HPV), human T-lymphotropic virus type 1 (HTLV-1), or Kaposi's sarcoma herpesvirus (KSHV)], a fungal infection [e.g., wherein the fungus is selected from the group consisting of Cryptococcus neoformans, Candida albicans, Histoplasma capsulatum, Coccidioides immitis, Paracoccidioides brasiliensis brasiliensis, Aspergillus species, and Zygomycetes], bacterial infections [e.g., wherein the bacteria is selected from the group consisting of Listeria monocytogenes, Borrelia burgdorferi, Neisseria meningitidis, Streptococcus pneumoniae, Staphylococcus aureus, Haemophilus influenzae, Mycobacterium, and Brucella species], or parasitic infections.

14. The method of treatment 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, synucleinopathies (i.e., Lewy body dementia and multiple system atrophy), polyglutamine diseases including spinocerebellar ataxia, prion diseases, traumatic brain injury, chronic traumatic encephalopathy, stroke, and spinal cord injury].

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

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

18. A method of treatment according to any one of claims 1 to 8, wherein the neuroinflammation is associated with a disease or condition selected from the group consisting of postoperative cognitive dysfunction, ischemia, brain tumors, chronic pain, headaches, migraines, trigeminal neuralgia, cardiovascular disease and related risk factors (e.g., due to obesity, hypertension, diabetes, hyperlipidemia, smoking, etc.), chronic peripheral inflammatory disorders (e.g., rheumatoid arthritis, inflammatory bowel disease, psoriasis, etc.), intestinal microbiota disturbances (i.e., imbalances), and herpetic keratitis.

19. A method of treatment according to any one of claims 1 to 8, wherein the neuroinflammation is caused by chemical intervention to treat the underlying condition or disease (e.g., use of TNF inhibitors to treat autoimmune diseases such as rheumatoid arthritis).

20. The method of treatment of any one of claims 1 to 8, wherein the neuroinflammation is caused by a mechanical intervention to treat the underlying condition or disease.

21. The method of treatment of any one of claims 1 to 8, wherein the neuroinflammation is caused by surgical intervention to treat an underlying condition or disease.

22. The method of claim 21, wherein the surgical intervention is selected from the group consisting of carotid endarterectomy, carotid angioplasty with or without stents, vertebral angioplasty with or without stents, intracranial angioplasty with or without stents, aneurysm coil embolization, vascular malformation occlusion, intracranial fistula repair, thrombectomy, extracranial-intracranial bypass, open heart surgery, heart surgery requiring a bypass machine, coronary artery bypass grafting, coronary angioplasty with or without stents, valve surgery / valvuloplasty, valvulotomy, patent foramen ovale closure, and great vessel surgery 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. The method of treatment according to claim 23, wherein the pharmaceutical composition further comprises a pharmaceutically acceptable carrier or excipient.

25. The method of treatment according to claim 23, wherein the pharmaceutical composition is composition MLC901.

26. The method of treatment according to claim 23, wherein the pharmaceutical composition is composition MLC1501.

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

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

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

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

32. The method of claim 30, wherein the test shows 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. A method of preventing or limiting neuroinflammation in a subject at risk of neuroinflammation due to injury, the method comprising administering to the subject an effective amount of an herbal composition comprising at least four herbal components: Polygala tenuifolia (root of Polygala), Astragalus membranaceus (root of Astragalus), Ligusticum chuanxiong rhizome, and Angelica dahurica (root of Angelica sinensis), or extracts thereof.

34. The method of claim 33, wherein the herbal composition further comprises at least one herbal component selected from the group consisting of Salvia miltiorrhiza root and rhizome (root of Salvia miltiorrhiza), Paeonia rubra root (root of Paeonia rubra), Carthamus tinctorius flower, Prunus persica (peach seed) and Acorus calamus rhizome (rhizome of Acorus calamus), or extracts thereof.

35. The method of claim 33, wherein the herbal composition consists essentially of: At least four herbal components: Polygala tenuifolia root (Polygala tenuifolia root), Astragalus membranaceus root (Astragalus membranaceus root), Ligusticum chuanxiong rhizome and Danggui root (Angelica sinensis root), or extracts thereof.

36. The method of claim 33, wherein the herbal composition consists essentially of: At least nine herbal components: Polygala root (root of Polygala), Astragalus root (root of Astragalus), Ligusticum chuanxiong rhizome, Danggui root (root of Danggui), Danshen root and rhizome (root of Danshen), Paeonia lactiflora root (root of Paeonia lactiflora), Carthamus tinctorius flower, Prunus persica (seed of peach) and Acorus calamus rhizome (rhizome of Acorus calamus), or extracts thereof.

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

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

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

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

41. The method of 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. 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. The method of any one of claims 33 to 42, wherein the damage is caused by a chemical, mechanical or surgical intervention to treat the underlying condition or disease as described in any one of claims 19 to 22.

44. A method for treating a subject at risk of developing a disease or condition due to neuroinflammation, the method comprising administering to the subject an effective amount of an herbal composition to prevent or reduce the development of the disease or condition, the herbal composition comprising at least four herbal components: Polygala tenuifolia (root of Polygala), Astragalus membranaceus (root of Astragalus), Ligusticum chuanxiong rhizome and Angelica dahurica (root of Angelica sinensis), or extracts thereof.

45. The method of treatment according to claim 44, wherein the herbal composition further comprises at least one herbal component selected from the group consisting of Salvia miltiorrhiza root and rhizome (root of Salvia miltiorrhiza), Paeonia rubra root (root of Paeonia rubra), Carthamus tinctorius flower, Prunus persica (peach seed) and Acorus calamus rhizome (rhizome of Acorus calamus), or extracts thereof.

46. ​​The method of treatment of claim 44, wherein the herbal composition consists essentially of: At least four herbal components: Polygala tenuifolia root (Polygala tenuifolia root), Astragalus membranaceus root (Astragalus membranaceus root), Ligusticum chuanxiong rhizome and Danggui root (Angelica sinensis root), or extracts thereof.

47. The method of treatment of claim 44, wherein the herbal composition consists essentially of: At least nine herbal components: Polygala root (root of Polygala), Astragalus root (root of Astragalus), Ligusticum chuanxiong rhizome, Danggui root (root of Danggui), Danshen root and rhizome (root of Danshen), Paeonia lactiflora root (root of Paeonia lactiflora), Carthamus tinctorius flower, Prunus persica (seed of peach) and Acorus calamus rhizome (rhizome of Acorus 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. The method of treatment according to claim 48, wherein the pharmaceutical composition further comprises a pharmaceutically acceptable carrier or excipient.

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

51. The method of treatment according to claim 48, wherein the pharmaceutical composition is composition MLC1501.

52. The method of treatment of 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. The method of treatment of any one of claims 44 to 52, wherein the disease or disorder is brain damage.

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

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

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

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

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

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

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