Treatment of traumatic encephalopathy by fibroblasts and therapeutic adjuvants

Abstract

To provide methods for treating neurological disorders by stimulating regenerative and anti-inflammatory activity of fibroblasts.SOLUTION: A method of increasing regenerative activity of a population of fibroblasts comprises exposing the population to one or more agents capable of inhibiting NF-κB. In specific cases, methods are utilized to treat or prevent central nervous system injury, such as chronic injuries including chronic traumatic encephalopathy.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] (Reference to Related Application) This application was filed on September 9, 2019, and is incorporated herein by reference in its entirety. This application claims priority to U.S. Provisional Application No. 62 / 897,429.

[0002] (Technical field) The disclosed embodiments are directed to at least one of the following fields: cell biology, molecular biology, neurology, physiology, biochemistry, It encompasses the fields of immunology and medicine. [Background technology]

[0003] Many pathologies are the result of inflammation, which is the body's response to injury or infection. The main events involved in the inflammatory process include the loss of blood supply to the site of injury or infection. Increased capillary permeability due to endothelial cell retraction; and increased capillary permeability due to increased blood flow from the capillaries to the periphery There is migration of leukocytes into tissues. Leukocytes enter the circulatory system partially due to increased capillary permeability. This allows larger molecules and cells to leave the cell that they normally cannot. This allows the passage of immune and leukocyte soluble mediators across the endothelium, White blood cells, primarily neutrophil polymorphonuclear (PMN) leukocytes, are able to reach the site of injury or infection. White blood cells (also known as neutrophils or PMNS) and macrophages undergo a process known as chemotaxis. At the site of inflammation, tissue damage and complement activation induce C5a Complement activation products also induce the release of chemotactic peptides such as phagocytes, mast cells, and It causes degranulation of leukocytes and basophils, smooth muscle contraction, and increased vascular permeability. The process by which inflammation and immune responses move from the bloodstream to the extravascular space involves a complex but coordinated series of events. At extravascular sites of infection or tissue injury, leukocytes and / or endothelial cells Bacterial endotoxins, which activate and cause one or both of these cell types to become adhesive, Antibody fragments of complement or interleukin-1 (DL-1), interleukin-6 (IL-6) Signals such as proinflammatory cytokines, including tumor necrosis factor (TNF), are produced. Initially, cells become temporarily adherent (evident by rolling), and then Afterwards, such cells become firmly adherent (evident by stickiness). Leukocytes migrate across the endothelial cell surface, diapedesis between endothelial cells, and enter the subendothelial matrix. Leukocytes migrate through the vascular network to the site of inflammation or immune response. Although leukocyte-endothelial interactions are necessary for host defense against foreign antigens and organisms, leukocyte-endothelial interactions are often This can have detrimental consequences for the host, for example, by inhibiting leukocyte adhesion and migration across endothelial cells. The globules release oxidants, proteolytic enzymes, and cytokines, which damage the endothelial cells. They directly damage the endothelial cells or cause functional damage to them. Leukocytes contribute to further tissue damage by releasing various inflammatory mediators. Furthermore, single leukocytes adhering within the capillary lumen or leukocytes within larger vessels Aggregation of spheres causes microvascular obstruction and ischemia. Leukocyte-mediated vascular and tissue injury occurs in acute and chronic allograft rejection, vasculitis, rheumatoid and other forms of inflammatory-based arthritis, inflammation skin diseases, adult respiratory distress syndrome, myocardial infarction, shock, stroke, organ transplants, crush injury and It has been implicated in the pathogenesis of a wide variety of clinical disorders, including ischemia-reperfusion syndromes such as limb retransplantation.

[0004] Many other serious clinical conditions involve underlying inflammatory processes in humans, e.g., multiple sclerosis MS (Multiple Skeletons and Multiple Myeloma) is an inflammatory disease of the central nervous system. In MS, circulating white blood cells cause inflammation. It infiltrates the damaged cerebral endothelium and damages the myelin, resulting in impaired nerve conduction and paralysis. In the case of concussion, these head injuries cause a buildup of damage that triggers inflammation.

[0005] A variety of anti-inflammatory drugs are currently available for use in treating conditions involving an underlying inflammatory process. However, their effectiveness varies widely and remains significant. There is a significant unmet clinical need for treatments with limited or no efficacy. Therapies available include those with unrelated side effects, such as: The present disclosure provides solutions to these problems. Summary of the Invention

[0006] administration of fibroblasts and minocycline and / or its analogs to the recipient Useful for stimulating regenerative and / or anti-inflammatory activity in cells, tissues, and / or organs of animals. Means, methods, and compositions of matter are disclosed. In one embodiment, fibroblasts are to treat or prevent a menstrual disorder, wherein the fibroblasts are administered in combination with minocycline and and / or analogs thereof, and combinations thereof with one or more regenerative cytokines to reduce inflammatory activity. At least some amount in the recipient effective to enhance the ability to induce production In some embodiments, minocycline (and The combination of fibroblasts with hydroxybenzoates and / or their analogs provides a therapeutic benefit for acute central nervous system injury such as stroke. In other embodiments, minocycline and / or analogs thereof are administered subsequently. For example, together with fibroblasts for the treatment of chronic injuries such as chronic traumatic encephalopathy (CTE). is administered.

[0007] The foregoing has outlined the features and technical advantages of the present disclosure in order that the detailed description that follows may be better understood. Those points have been outlined rather broadly. Additional features and advantages which form the subject of the claims of this specification. The disclosed concepts and specific embodiments are intended to carry out the same objectives of the present design. Those skilled in the art will recognize that the present invention may be readily utilized as a basis for modifying or designing other structures to achieve the same. It should be understood by those skilled in the art that such equivalent constructions are within the scope of the appended claims. It should be understood by those skilled in the art that the present invention does not depart from the spirit and scope of the invention as described herein. The novel features believed characteristic of the designs disclosed herein provide further objectives and The following claims, both as to construction and method of operation, together with their advantages, should be considered in connection with the accompanying drawings. However, the figures are for illustrative and descriptive purposes only and will be better understood from the following description. It should be expressly understood that these figures are provided for purposes of illustration only and are not intended as a definition of the limits of the present disclosure. It is. [Brief explanation of the drawings]

[0008] For a more complete understanding of the present disclosure, reference is now made to the following descriptions taken in conjunction with the accompanying drawings, in which:

[0009] [Figure 1] Figure 1 shows the synergistic effect of fibroblasts and minocycline in suppressing inflammation as measured by IL-1β. Bars from left to right are control, fibroblasts, minocycline, and the combination of fibroblasts and minocycline.

[0010] [Figure 2] Figure 2 shows the synergistic effect of fibroblasts and minocycline in suppressing inflammation as measured by TNF-α. Bars from left to right are control, fibroblasts, minocycline, and the combination of fibroblasts and minocycline.

[0011] [Figure 3] Figure 3 shows the synergistic effect of fibroblasts and minocycline in suppressing inflammation as measured by IL-6. Bars from left to right are control, fibroblasts, minocycline, and the combination of fibroblasts and minocycline.

[0012] [Figure 4] Figure 4 demonstrates that fibroblasts enhance the ability of minocycline to induce T regulatory cells. DETAILED DESCRIPTION OF THE INVENTION

[0013] I. Definition

[0014] Unless otherwise defined, all technical and scientific terms used herein are defined by the principles of the present disclosure. The terms "terms" and "terms" have the same meaning as commonly understood by one of ordinary skill in the art to which they pertain. Any methods and materials similar or equivalent to those described herein may be used in the practice or testing of this disclosure. Although any method and material may be used, preferred methods and materials are described. The nomenclature utilized in connection with, and techniques for, chemical synthesis and synthesis is well known in the art. , which are commonly used. Specific experimental techniques not specifically defined are generally , according to conventional methods well known in the art and as cited and discussed throughout this specification. This is done as described in various general and more specific references. To facilitate this discussion, the following terms are defined below:

[0015] The term "neuroprotection" refers to the ability to reduce, stop, or ameliorate neuronal damage; Protective and regenerative for nerve tissues affected by nerve damage, such as in suspected neurodegenerative diseases Alzheimer's disease (AD), age-related memory impairment, This includes the reduction of neuronal cell death or loss of function in diseases such as mild cognitive impairment and vascular dementia. The term may be used in conjunction with known methods, including biomarkers, PET imaging, etc. These neurodegenerative diseases are associated with neurodegenerative disorders that can be diagnosed by For an example of how to determine ent effects in Alzheimer's and other neu "rodogenic diseases by MRI and MRS" See NMR Biomed 2006 Oct;19(6):655-668.

[0016] A "pharmaceutically acceptable" excipient is one that does not cause excessive harmful side effects commensurate with a reasonable benefit / risk ratio. can be used with humans and / or animals without adverse reactions (such as toxicity, irritation, and allergic reactions) It is a suitable excipient for use in

[0017] A "safe and effective amount," as used in the present invention, is a dose that is consistent with a reasonable benefit / risk ratio. The desired treatment can be achieved without undue adverse side effects (e.g., toxicity, irritation, or allergic response) associated with the administration of the compound. The amount of a component sufficient to produce a therapeutic response.

[0018] In some embodiments, a "therapeutically effective amount" is an amount effective to produce a desired therapeutic response. A safe and effective amount of the ingredients, for example, to prevent or treat neurodegeneration, memory loss, and / or dementia. It refers to the amount that is effective to improve the condition.

[0019] "Allogeneic," as used herein, refers to cells of the same species that are genetically distinct from the host's cells. refers to cells.

[0020] As used herein, "autologous" refers to cells derived from the same subject. The term "graft" as used herein refers to the transfer of tissue to the tissue of interest in vivo through contact with existing cells of the tissue. Refers to the process of incorporating stem cells into tissue.

[0021] "Approximately" or "about" as used herein applies to one or more values of interest. When used, the term "approximately" or "about" refers to a value similar to the stated reference value. In this embodiment, the term "approximately" or "about" is used unless otherwise specified or clear from the context. Unless otherwise specified, the values are in either direction (greater or less) of the stated reference value. 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 1 2%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or A range of numbers that falls within a smaller range (provided that the number does not represent 100% of the possible values). This does not apply if it exceeds the limit.)

[0022] "Carrier" or diluent: As used herein, the terms "carrier" and "diluent" refer to a pharmaceutical Pharmaceutically acceptable (e.g., safe and non-toxic for human administration) compounds useful in preparing formulations ) refers to a carrier or diluent. Exemplary diluents include sterile water, bacteriostatic water for injection (BWFI), pH buffer solutions (e.g., phosphate buffered saline), sterile saline, Ringer's solution, or Dextromethorphan Contains Strohs solution.

[0023] Dosage Form: As used herein, the terms "dosage form" and "unit dosage form" refer to a dosage form that is administered to a patient to be treated. Each unit is a physically discrete unit of a therapeutic agent for producing a desired therapeutic effect. However, the total dosage of the composition may vary depending on the individual patient. It will be understood that such decisions will be made by the attending physician within the scope of sound medical judgment.

[0024] Dosing regimen: A "dosing regimen" (or "therapeutic regimen") is a medical treatment regimen as that term is used herein. When used, unit doses administered individually to a subject, typically separated by a period of time In some embodiments, a given therapeutic agent is a set of (typically two or more) In some embodiments, the dosage is If the regimen includes multiple doses, each separated from each other by the same length of time, In some embodiments, the dosing regimen may include multiple doses and at least two intervals separating the individual doses. In some embodiments, the therapeutic agent is administered continuously for a predetermined period of time. In some embodiments, the therapeutic agent is administered once daily (QD) or twice daily (BID). ) is administered.

[0025] The term "culture-expanded population" refers to cells whose numbers have been increased by cell division in vitro. This term applies equally to stem cell populations and non-stem cell populations, including fibroblasts. It can be used.

[0026] The term "passaging" refers to the process of transferring a portion of cells from one culture vessel to a new one. vinegar.

[0027] The term "cryopreservation" refers to the preservation of cells for long-term storage in a cryoprotectant at low temperatures. Refers to...

[0028] The term "master cell bank" refers to a collection of cryopreserved cells. The bank may contain fibroblasts, stem cells, non-stem cells, and / or a mixture of stem and non-stem cells. Any cells may be obtained from a master cell bank and / or may be deposited in the

[0029] The disclosure provides a method for the production of T regulatory cells using minocycline as a means of activating T regulatory cells. Stimulating and / or inducing proliferation of and / or inducing de novo production of These include means to "program" the immune system to suppress autoimmunity via

[0030] II. Embodiment

[0031] The present disclosure provides a method for the treatment of any animal, including humans, dogs, cats, horses, etc., from injury and / or disease. The present invention encompasses the treatment and prevention of brain-related medical conditions, including those for mammals. The disclosed methods treat or prevent neurological damage. The medical condition may be a neurological disorder. It has the potential to treat or prevent all types of brain injury, including traumatic brain injury. Injuries may include hematoma, hemorrhage, concussion, edema, or a mixture of these. Types of Traumatic Brain Injury Types include cerebral contusion, second impact syndrome, Coup-Contrecoup brain injury, Shaken baby syndrome and / or penetrating injury.

[0032] A medical condition may be the result of a single injury or repeated injuries. Harm may be due to physical contact, including as a result of work and / or sports. The condition may be a neurological disorder. Any injury may be present before any symptoms appear. It could have occurred at any time during an individual's life, including years, months, days, or weeks. In certain embodiments, the medical condition is chronic traumatic encephalopathy (CTE), which includes Pugilistica dementia is included. Individuals who play soccer, boxing, Recreational sports include wrestling, soccer, hockey, lacrosse, and basketball. The individual may be an athlete, including those involved in sports or professional activities. Construction workers, first responders, warehouse workers, and others who have suffered medical injuries such as head injuries related to their jobs It may have a condition.

[0033] Individuals should take precautions before being exposed to environments that pose a risk of head injury, such as sports stadiums, construction sites, etc. The disclosed treatments may be provided, whether before and / or after a head injury. Any administration of the therapy may be a single or multiple administrations. Any duration between doses may be utilized, including on the order of several years. Treatment is given within 1 to 60 minutes of injury, within 1 to 24 hours of injury, within 1 to 4 weeks of injury, Provided to individuals within 1 to 12 months of injury or within 2 years or more after injury.

[0034] In some cases, the fibroblasts are exposed to one or more anti-inflammatory agents prior to use to enhance the therapeutic activity of the fibroblasts. In certain embodiments, one or more anti-inflammatory agents (e.g., NF-κB inhibitors) are co-administered with the fibroblasts to enhance the therapeutic activity of the fibroblasts. In one embodiment, the administered fibroblasts are allogeneic fibroblasts, and the co-administered anti-inflammatory agent is minocycline and / or its analogs (e.g., tigecycline). Any of the compositions encompassed herein can be provided to individuals at risk of head injury, such as before playing sports or entering a dangerous work environment.

[0035] In one aspect, the disclosure provides a method for treating a cancer cell line comprising administering to a patient a therapeutically effective amount of at least one compound selected from the group consisting of fibroblasts and minocycline (and / or an analog thereof), and and / or low-dose interleukin-2 and one or more other immunomodulatory agents. Administration of a combination of cells was used to generate stable regulatory T cells ( This disclosure deals, in part, with the differentiation of T cells into Tregs (Tregs). Administration of α-glucan analogs has been shown to be effective in healthy animals and animals suffering from neurological problems such as chronic traumatic encephalopathy. This is based on the observation that the IL-16 receptor agonist has the ability to regulate the number of T regulatory cells. The authors report the use of minocycline to stimulate T regulatory cells, as well as the expansion of T regulatory cells. The results, detailed later, showed that minocycline induces significant immunosuppressive activity. Instead, it induces a state of "active immune tolerance." Indeed, T regulatory cells play a key role in the progression of remission after cessation of therapeutic drugs. Based on the fact that tolerance-inducing processes can induce a state of "infectious tolerance" that maintains itself Therefore, it is reasonable to consider that the present findings are significantly different from previous findings. Such maintenance of T regulatory cells has been previously described by numerous researchers [1-4]. Harnessing the newly discovered properties of minocycline to stimulate cells could be used to treat a wide variety of diseases. As the utility of this approach becomes apparent, it is possible to develop therapeutic compounds such as One likely advantage to the use of steroids is reduced tolerance (e.g., decreased steroid activity) in human (or other) subjects. As used herein, the term "nicotine" refers to an increased risk of side effects (e.g., increased toxicity and side effects). "Potential T cells" are lymphocytes that typically originate from the thymus and express the T cell receptor. Naive T cells typically undergo basic development in the bone marrow and then undergo further differentiation into positive and negative T cells in the thymus. However, naive T cells have not yet encountered their cognate antigen in the periphery. The terms "activate" and / or "differentiate" refer to the process by which naive T cells undergo differentiation in vivo. At least four different T cell types characterized by distinct expression profiles and functions It refers to the process of inducing one of the lineages to develop further. " and / or "differentiated" means that it is identified as a specific activated / differentiated lineage. As can be seen, previously naive cells are now induced to express specific genes. The term can also refer to the cell that develops as a result of cell division into many progeny cells. These cells are produced from proliferation into cells and carry identifiable markers for specific activation / differentiation lineages. Therefore, "activating naive T cells" can refer to cells that of differentiated T cells from the initial naive T cells as well as the initial T cells after transcription is induced It can refer to the production of an expanded cell population.

[0036] For any embodiment, a method for producing naive T cells activated into the desired differentiated T cells is provided. The minimum amount of minocycline (and / or its analogs) required for It is possible.

[0037] In one embodiment, minocycline (and / or its analogs) is administered to naive T To treat naive T cells differentiated into cells with increased FoxP3 expression compared to normal T cells, The term "FoxP3" refers to "forkhead box P3" or "skeleton box P3." The FoxP3 protein is a transcription factor that regulates transcription. It belongs to the head / winged helix family. In regulatory T cell model systems, FoxP3 occupies the promoters of genes involved in regulatory T cell function and regulates T cell receptor activa- tion It may suppress the transcription of important genes following somatic stimulation. Therefore, FoxP3 Regulatory T cells are involved in peripheral antigen tolerance and generally promote defense against inflammatory responses. FoxP3 protein is known to be a master regulator in the development of Treg cells. Examples of proteins include human (Entrez#:50943; RefSeq(mRNA): NM_001114377; RefSeq (amino acid): NP_001107849) and and mouse (Entrez#:20371; RefSeq(mRNA):NM_00119 9347; RefSeq (amino acid): NP_001186276). Other FoxP3 proteins and gene homologs are known for vertebrates, and their Expression can be easily determined. As used herein, the term "increase" refers to the initial increase in expression during differentiation. derived from the same individual (or individual of the same species) as the original naive T cells Detectably greater than expression levels in naive T cells like other naive T cells The expression level of the FoxP3 transcription factor. Increased expression indicates the underlying foxp3 gene. Routine and established methods known in the art for determining levels of transcription or functional FoxP3 in offspring are used. The determination can be made using the methods described above.

[0038] In one embodiment, naive T cells are differentiated into T regulatory cells (Tregs). "e.g." refers to a lineage of T cells that promotes or maintains tolerance to an antigen, typically a self-antigen. Tregs were previously called "suppressor T cells," but Tregs are generally considered to be As mentioned above, Tr Treg cells are typically characterized by positive or increased expression of FoxP3. It is also characterized by additional positive or increased expression of CD4 and CD25. Thus, in one embodiment, Tregs are CD4+, CD25+ and FoxP3+ expressing Tregs. It is characterized by:

[0039] In another embodiment, the contact of naive T cells induces "Th17" inflammation by differentiated T cells. For example, contact of naive T cells results in inhibition of RORγT expression or This results in a decrease in the Th17 (pro-inflammatory) phenotype of activated T cells normally involved in mucosal immunity. ) is a phenotypic marker, and in one embodiment herein, naive T cells are cultured The contact may be in vitro in a culture medium. Typically, the culture medium supports T cell viability and The medium also contains factors that are generally known to promote differentiation into the desired lineage. The antibody may contain additional components known to promote T cell activation toward the immune system. Such additional components are often called "distortion" components. Distortion factors can also be present in other microorganisms. Bioflora metabolites (e.g., short-chain fatty acids, bile acids, polysaccharide A), dietary compounds (e.g., n3 Polyunsaturated fatty acids, retinoic acid, and other vitamin derivatives (VitD, VitC, etc.) ), polyphenols, quercetin, resveratrol, NSAIDS, TGF-β, I Other distortion factors useful for this purpose include IL-10, rapamycin, and IL-2. , as known in the art, curcumin, metformin / AMPK activators The present invention includes agonists, PI3 kinase / Akt inhibitors, and PPAR agonists. reported that minocycline inhibits the ability of a "skewing factor" to generate enhanced numbers of T regulatory cells. It teaches increasing force.

[0040] In another embodiment, the present disclosure provides a method of producing Treg cells. In one embodiment, the method includes contacting naive T cells in vitro with minocycline. wherein the minocycline is, as described above, a component of a standard culture medium (e.g., naive T cells can be contacted with the Tregs as an additive (additive). This method allows for the differentiation of Tregs into their differentiated Treg state. The method may include further culturing and / or expanding the activated T cells in a culture medium.

[0041] As described below, the inventors have demonstrated in vivo efficacy using the disclosed minocycline. In vitro induced Tregs ("iTregs") are induced Tregs generated using existing technology. We have demonstrated that iTregs have novel characteristics compared to iTregs. showed that iTregs arising from the application of TDMMs such as indole are in a “pro-inflammatory” environment. We demonstrated that this generated stable iTregs that did not revert to the Th17 phenotype even after treatment with iTregs. In another aspect, the present disclosure provides induced T regulatory cells (iTregs). e.g., is produced by the methods described herein. In some embodiments, iT Reg is generated by contacting naive T cells with minocycline. Reg is the first T cell after activation or one or more cell divisions after the first T cell. In some embodiments, the progeny cells may be in a differentiated state after expansion has occurred. iTregs are more likely to commit to the Treg lineage compared to iTregs induced using conventional means. For example, IL-4, IL-6, and IL-23 all exhibit increased stability in This impairment is known to reduce the stability of typical Tregs. Therefore, the iTreg lineage is regulated by IL-4, IL-6, and IL-23. In some embodiments, iTregs are less susceptible to induced instability due to CT. LA4, CD62L, CD25, higher Foxp3, α4β7, and / or CCR9 The relative increase in expression distinguishes them from typical Tregs, which can be detected by routine testing. can be easily determined.

[0042] In another aspect, the present disclosure provides a method for treating rheumatoid arthritis by administration of minocycline (and / or analogs thereof). In a specific embodiment, this provides a method for increasing the stability of Treg cells. Proinflammatory cytokines and signaling, e.g., IL-4, IL-6, and IL-2 3, etc., which alters the Treg-specific expression profile, resulting in decreased sensitivity of Tregs. Treg cells can be generated by the novel methods described herein or by methods already known in the art. The Tregs may be induced Tregs (iTregs), such as those produced by existing methods. Alternatively, Tregs can be naturally occurring Tregs (nTregs). "Tregs" refers to Tregs that exist in vivo without prior in vitro intervention or transfer, and typically Typically, the thymus is obtained from a human. This method involves the use of a Tre The Treg population was isolated and the Treg population was already ex vivo expanded and Tregs were then isolated. Exposure to nocycline, or a precursor, prodrug, analog, or acceptable salt thereof In some cases, the target population may be subjected to in vitro , iTreg populations produced by the novel methods described herein, Tregs are those already exposed to minocycline, or its precursors, prodrugs, or acceptable salts. and may or may not have further exposures.

[0043] In another aspect, the present disclosure provides a method for reducing, preventing, ameliorating, or reducing inflammation in a subject in need thereof. and / or methods for treating inflammation-related diseases. One method for using isolated or ex vivo / in vivo differentiated Treg cells as part of a recombinant T cell therapy The method of this embodiment is the same as that described immediately above, i.e., the method of using naive T cells. iTr produced by contacting In some embodiments, the subject is treated with an anti-inflammatory drug, such as steroids, to treat excessive or harmful inflammation. In some embodiments, the subject is suffering from or susceptible to an allergic disease. Inflammatory bowel disease, colitis, NSAID enteropathy / ulceration, psoriasis, rheumatism, graft vs. Host disease, lupus, multiple sclerosis, etc., or susceptible to them. In some embodiments, the subject has a tumor that is a target of indole, such as mTor, stat3, or akt characterized by the role of erk, jnk, stat5, and / or smad2 / 3 Additionally or alternatively, the subject has or is susceptible to a disease. They can suffer from harmful inflammation due to cancer or infection from microbial or parasitic pathogens. The e.g., may be formulated for administration by any suitable route according to known standards and methods. For example, iTregs can be administered intraperitoneally (IP), intravenously (IV), topically, parenterally, intradermally, or transdermally. , orally (e.g., via liquid or pill), inhaled (e.g., intranasal mist), and other suitable In some embodiments, administration is to a mucosal area of a subject. (e.g., direct administration into the digestive tract).

[0044] In some embodiments, the method comprises administering in vivo Tr as described herein. In one such embodiment, the subject is administered an effective amount of minocycline. administration of cyclohexyl 1,2-diol, or a precursor, prodrug, analog, or acceptable salt thereof Minocycline (and / or its analogs) can be administered via any suitable route. For example, minocycline and / or analogs may be administered intraperitoneally (IP), intravenously (IV), or intravenously. (IV), topical, parenteral, intradermal, transdermal, oral (e.g., via liquid or pill), rectal, or by the respiratory (e.g., intranasal mist) route. Therefore, minocycline can be taken, for example, via a liquid or pill, and Facilitates delivery to the intestinal tract.

[0045] In some embodiments of the present disclosure, a drug (e.g., minocycline or minocycline) phosphorus prodrugs) are administered systemically to achieve therapeutically effective plasma concentrations in patients. However, oral drug dosage forms, including those containing minocycline, are therapeutically delivered. To achieve effective systemic concentrations, several obstacles must be overcome. Tetracyclines are generally highly lipophilic. Their limited water solubility is due to This limits the amount of tetracycline available for absorption in the gastrointestinal tract. Minocycline, like other tetracyclines, is substantially absorbed from the human gastrointestinal tract. Finally, if patients avoid taking oral medications or if oral dosage forms are unable to deliver the full dose, and do not remain in the gastrointestinal tract long enough to release and achieve therapeutic concentrations, causing the patient to experience nausea or If vomiting occurs, the oral bioavailability of any product will be further reduced. From the above viewpoint, it is desirable to administer the drug to the pancreas via an administration route that does not depend on absorption from the gastrointestinal tract of mammals. One or more medical conditions that respond to tetracyclines, including pancreatic cancer, pancreatitis, pain, nausea, or appetite stimulation For the treatment of certain conditions, a therapeutically effective amount of a tetracycline (e.g., minocycline or minocycline) is administered. It is desirable to deliver a compound (a cyclohexyl 1-hydroxybenzoate prodrug) systemically to a mammal in need thereof. One parenteral route of administration for systemic delivery of minocycline is transdermal administration. In addition, the epidermis and dermis of many mammals, such as humans and guinea pigs, are permeable to the stratum corneum. Contains enzymes capable of metabolizing the active agent in the skin of mammals, such as humans. The resulting metabolic processes result in the delivery of pharmaceutically effective amounts of the compound into the systemic circulation of mammals in need thereof. It can be utilized to deliver tracyclines (e.g., minocycline). Described in the literature are prodrugs of tetracyclines (e.g., minocycline prodrugs). and tetracycline prodrugs that can be administered transdermally to mammals (e.g., humans). The composition contains a drug, so that the metabolites resulting from metabolism in the skin are tetrahydrofuran. Systemically for the treatment of cyclin-responsive medical conditions (e.g., pancreatitis and pancreatic cancer) Unfortunately, due to its highly lipophilic nature, Iclin is hardly absorbed through membranes such as the skin of mammals, including humans. Thus, a therapeutically effective amount of minocycline can be administered to a mammal in need thereof within a reasonable time frame. However, the success of transdermal administration over an adequate surface area is substantially limited.

[0046] The use of minocycline for the suppression of inflammation has been previously described in the art. The present invention provides a means to utilize the anti-inflammatory effects of minocycline for the stimulation of Treg cells. Once Treg cells are generated, the present invention provides a method for expanding such Treg cells. This teaches that it can be done.

[0047] The present invention provides a means of utilizing minocycline to prevent unwanted immune responses. For example, in pregnancy, "tolerogenic antigen presentation" occurs only via the indirect pathway of antigen presentation. [5] Another pathway for selective tolerogen production in pregnancy is the stimulation of Treg cells. , which has been shown to be essential for successful pregnancy [6]. In this study, to generate antigen-presenting cells from fibroblasts, transfection with MHC or MHC-like molecules was performed. The present invention teaches the modification of fibroblasts by transfection, where antigen-presenting cells are used for therapeutic purposes. When administered to a host at a sufficient concentration and frequency, it can induce antigen-specific tolerance. The series shows depletion of tumor-specific T cells, while T cells with specificity for other antigens It has been shown that tumors and tumor-associated cells are spared this risk [7-1 0]. This allows cancer to make the host generally immune responsive, while This is why Treg cells can selectively induce "Lee's holes." Furthermore, Treg cells are likely involved in tumor immunity. It has been demonstrated that the immune system actively suppresses anti-tumor T cells as a "backup" mechanism for immune evasion. At the clinical level, tumors are known to have a prognostic value due to their ability to inhibit peripheral T cell activity [11-13]. Many studies have linked this to poor performance [14-16]. Therefore, in one embodiment of the present disclosure, , the use of molecules that stimulate the generation of Tregs, as well as molecules that expand the generated Tregs. In one embodiment, fibroblasts are administered to enhance Treg generation. In addition, the cells are transfected with one or more autoantigens along with interleukin-2. In some embodiments, interleukin-2 is used to enhance the in vivo expansion of Tregs. It is administered physically.

[0048] In one embodiment, tolerance is induced to an autoantigen that is part of CTE initiation and progression. The present inventors believe that CTE has an autoimmune component, and that suppressing this component may be beneficial for fibroblast therapy. We believe that this could promote the efficacy of the IFN-γ-γ receptor. A natural example of tolerance that is utilized by the present disclosure is oral tolerance. The detected antigen is then transformed into antigen-specific TGF-β-producing cells (sometimes called "Th3") [17- This process induces the production of immune cells

[19] and Treg cells [20, 21]. Ingestion of antigens containing antigen II

[22] specifically stimulates both T cells and B cells. It has been shown that the suppression of responses is induced [23,24]. Effector cell loss / anergy is associated with antigen presentation in a tolerogenic manner.

[0025] Oral administration of autoantigens has been reported to induce disease remission in animal models of RA

[26] , multiple sclerosis

[27] , and type 1 diabetes

[0028] . Furthermore, rheumatoid arthritis

[29] has been shown to be a potential therapeutic target for rheumatoid arthritis. , autoimmune uveitis

[30] , and multiple sclerosis

[31] . Clinical trials have shown the efficacy of oral immune tolerance. Common molecules and mechanisms are at work in both. Therefore, natural means of inducing tolerance are Tolerogenicity, which produces molecules similar to those found in the physiological state of tolerance induction. In some embodiments, oral tolerance is achieved by administering "universal donor" cells containing The autoantigen-transfected fibroblasts of the invention can be used together, for example, in the treatment of type 1 diabetes. If the patient is treated, the patient is given minocycline and a diabetic specific drug such as GAD65. In one embodiment, cells transfected with a specific autoantigen are administered, and the cells express IL-10. The cells may be transfected with a tolerogenic molecule such as α-tolerogenic β ... Orally delivered GAD65 may be utilized to enhance the inflammatory process. The present invention provides an autoantigen combined with a molecule involved in oral tolerance development, such as TGF-β. This paper teaches transfection of cells by

[0049] The present disclosure provides evidence that administration of minocycline and other compounds associated with inhibiting inflammation can enhance the ability of fibroblasts to This includes the previously unexpected finding that it can potently enhance regenerative activity. In some embodiments, the dose of minocycline is determined based on the individual's needs, the individual's condition, and adjusted based on the underlying disease. 10-100 mg (or approximately 1 mg-400 mg, About 10mg-300mg, about 10mg-150mg, about 10mg-120mg, about 10m g-100mg, about 20mg-400mg, about 20mg-300mg, about 20mg-20 0mg, about 30mg-400mg, about 30mg-300mg, about 30mg-200mg, About 30mg-100mg, about 50mg-400mg, about 50mg-300mg, about 50m g-200mg, about 50mg-100mg, about 10mg-90mg, about 10mg-80m g, about 10mg-70mg, about 10mg-60mg, about 10mg-50mg, etc.) Icrin and about 10-400 mg (e.g., about 50-200 mg, about 10-300 mg, about 10-200mg, about 10-150mg, about 10-100mg, about 10-90mg, about 10-80mg, approximately 10-70, approximately 10-60, approximately 10-50mg, approximately 20-400mg , about 20-300, about 20-200mg, about 20-100mg, about 20-90mg, about 3 0-500mg, about 30-400mg, about 30-300mg, about 30-200mg, about 3 0-100mg, etc., about 40-500mg, about 40-400mg, about 40-300mg, about 40-200mg, about 40-100mg, about 50-500mg, about 50-400mg, about Between minocycline (50-300 mg, approximately 50-100 mg, approximately 50-80 mg, etc.) A variety of dosages can be used, including:

[0050] In one embodiment of the present disclosure, minocycline has therapeutically relevant activity These drugs are used based on their properties known in the art. For example, minocycline It has been shown that this antibiotic can inhibit activation of the Minocycline has been shown to protect hippocampal neurons against global ischemia in repts. showed that CA1 pyramidal neuron survival was 10.5% when treatment was initiated 12 hours before ischemia. The effect of doxycycline on ischemia was increased to 77% when treatment was initiated after 30 minutes of ischemia, and to 71% when treatment was initiated after 30 minutes of ischemia. The survival rates before and after treatment with minocycline were 57% and 47%, respectively. The α-aminobutyric acid (NADPH)-induced activation of microglia and the appearance of NADPH-diaphorase-reactive cells were also observed. However, induction of glial acidic fiber protein, a marker of astrogliosis, was not affected. Minocycline treatment for 4 days did not affect the activation of microglia after ischemia. 70% reduction in mRNA induction of the spase interleukin-1 beta converting enzyme Similarly, minocycline-treated animals showed significantly increased expression of inducible nitric oxide synthase mRNA. The protective effect of minocycline against inflammation-associated neuronal cell death was Minocycline (0.02 microm) is glutamic acid 500 microm or cain In mixed spinal cord (SC) cultures treated with 100 microm of phosphate for 24 hours, Treatment with these excitotoxins significantly increased survival in mice in separate studies. It induces dose-dependent microglial proliferation, which is associated with interleukin-1β (IL-1β) This was associated with increased release of ATP, followed by increased release of lactate dehydrogenase (LDH). The toxicity was enhanced when microglial cells were cultured on SC cultures. Excitotoxin-induced microglial proliferation and increased release of nitric oxide (NO) metabolites and IL-1β. Excitotoxins also induced microglial proliferation in pure microglial cultures and inhibited NO metabolism. These responses were inhibited by minocycline. In both SC and pure microglial cultures, excitotoxins induce p38 mitogen activation exclusively in microglia. Minocycline activated p38 MAPK in SC cultures. Inhibits p38 MAPK activation in the culture medium and inhibits the p38 MAPK inhibitor SB Treatment with 203580 increased neuronal survival but not p44 / 42 MAPK inhibitors. The inhibitor PD98059 did not increase the expression of guanylate β-glucan. Glutamate induces transient activation of p38 MAPK, which is suppressed by minocycline.

[33] One interesting feature of minocycline is that it can be used at low concentrations. One study showed that nanomolar concentrations of minocycline inhibited neurons in mixed spinal cord cultures. It has been shown that NMDA treatment alone protects against NMDA excitotoxicity. Microglial proliferation was induced by the addition of extra microglial cells onto these cultures. Minocycline abolished these responses to NMDA. Minocycline also inhibited NMDA-induced neurite outgrowth in pure microglial cultures. It suppressed the proliferation of microglial cells and the increased release of IL-1β and nitric oxide caused by inflammatory bowel disease. Furthermore, minocycline inhibits the p38 mitogen-activated protein kinase activity in microglial cells. inhibits NMDA-induced activation of MAPK, and p44 / 42 MAPK inhibitors A specific p38 MAPK inhibitor reduced NMDA toxicity

[32] .

[0051] In one embodiment of the present disclosure, minocycline is activated by activated microglia. The fibroblasts induce a therapeutic effect on the brain in the absence of chronic inflammation induced by the To enable this, microglial activation may be induced before, simultaneously with, or after administration of fibroblasts. This is because microglial activation has been shown to be a major cause of It is useful in conditions such as CTE, which has been shown to be effective in various aspects of CTE, such as depression. It has been shown to be associated with various pathological conditions.

[0052] In some embodiments, minocycline is administered to patients receiving fibroblasts. In this context, it is administered to modify the interaction between T cells and microglia [46 Modulation of T cell activity may be desirable to enhance the survival of allogeneic fibroblasts. Alternatively, modulation of T cell activity may be a trophic factor that enhances the activity of fibroblasts. It can be used to produce trophic factors.

[0053] The study of neuropathologies such as CTE has shown that many preclinical observations suggest that neuroprotective approaches may also be beneficial. The majority of evidence strongly suggests that it may have an effect on cell death. This suggests that several pathological pathways leading to Simultaneous targeting of different pathways should be a rational approach to treatment. In one embodiment of the present disclosure, minocillin, an anti-inflammatory drug that blocks microglial activation, is used. Antibacterial agents with apoptotic and anti-inflammatory properties, riluzole, glutamate antagonists and A three-drug cocktail consisting of nimodipine, a voltage-dependent calcium channel blocker, was used. Evidence that fibroblast activity can be increased by ATP in a mouse model of amyotrophic lateral sclerosis (AMLS) It is provided herein that the compound exerted significant neuroprotection in

[0054] For use within the scope of the present disclosure, minocycline is a semisynthetic tetracycline derivative. It is widely used in humans because it effectively crosses the blood-brain barrier and has relatively few side effects. Minocycline prevents microglial activation and reduces the induction of caspase-1. This reduces the levels of the mature inflammatory cytokine IL-1β and promotes mitochondrial It is suggested that the inhibitory effect of cytochrome c release from α-glucan is neuroprotective. [59-68]. Furthermore, minocycline, doxycycline, and their non- Antibiotic derivatives (chemically modified tetracyclines) inhibit matrix metalloproteinases, Nitric oxide synthase, protein tyrosine nitration, cyclooxygenase-2 and pro It has been shown to inhibit staglandin E2 production. Recent studies performed with glial cultures have shown that minocycline inhibits excitotoxin-induced microenvironmental responses. It was demonstrated that it may also have neuroprotective effects through inhibition of glial activation. Icrin mediates glutamate and kinase activity of p38 MAPK, which is activated only in microglia. In some embodiments of the present invention, minocycline inhibits niacin-induced activation. , and antiglutaminergic drugs such as riluzole, a glutamate antagonist. currently approved for the treatment of ALS with only a modest effect on survival. It is the only drug that can 1) N.Eng.J.MED. 344,1688-1699) In two controlled clinical trials Riluzole has been shown to extend the survival of ALS patients by 3 to 6 months. The exact mechanism of action of riluzole is unclear. The mechanism is unclear, but it is likely due to inhibition of glutamate release and blockage of sodium channels. through activation or inactivation and / or activation of G protein-coupled transduction pathways in the CNS. The disruption of excitatory amino acids (EAA) is thought to be involved. When tested as monotherapy in Increased survival (Gurney, ME, et al. (1996) Ann. Neurol. . 39,147-157).

[0055] In some embodiments of the present disclosure, minocycline induces tolerogenic dendritic cells in vivo. 69], and tolerogenic dendritic cells are used to induce T regulatory cells. The T regulatory cells are used to suppress inflammation and enhance the activity of transplanted fibroblasts. do.

[0056] For administration of minocycline or a derivative thereof, in some embodiments, the present disclosure When a drug of the present disclosure or a combination drug of a drug of the present disclosure and another drug is used for the above purpose, It is generally administered systemically or locally in oral or parenteral form.

[0057] The dosage varies depending on age, weight, symptoms, therapeutic effect, administration method, treatment period, etc. Usually, the dose is 1 to 100 mg per adult, administered orally once or several times a day, or The dose ranges from 0.1 to 50 mg per person, and can be administered once or several times a day, once or several times a week, or in the form of a sustained-release preparation. Parenteral administration: once or several times every three months, or continuously administered intravenously for one to 24 hours a day Of course, the dose may vary depending on various conditions as mentioned above, so it may be necessary to administer a dose less than the above range. In some cases, a dose not exceeding the above range may be sufficient, and in other cases, it may be necessary to administer the drug in a dose exceeding the above range. When administering the drug of the present invention or a combination drug of the drug of the present invention with other drugs, the drug of the present invention is administered As internal solid or liquid formulations for oral administration, or as injections, subcutaneous or Used in intramuscular injections, topical preparations, suppositories, eye drops, inhalants, and medical device-containing preparations. Solid formulations for internal use for oral administration include tablets, pills, capsules, Powders, granules, etc. are included. Hard capsules and soft capsules are included in capsules. In such solid oral preparations, one or more active substances are used as such or in combination with a filler (e.g., lactose, mannitol, glucose, microcrystalline cellulose, starch, etc.), binder - (hydroxypropyl cellulose, polyvinylpyrrolidone, aluminometasilicate magnesium, etc.), digestive aids (calcium cellulose glycolate, etc.), lubricants (stearic acid, Magnesium phosphate, etc.), stabilizers, solubilizing aids (glutamic acid, aspartic acid, etc.) The mixture is mixed with other ingredients such as cereals, dairy products, and pharmaceutical preparations, and then used. This is coated with a coating agent (sucrose, gelatin, hydroxypropyl cellulose, hydrolyzed or coated with two or more It may contain a further capsule of an absorbent material such as gelatin. It can be enjoyed.

[0058] Liquid formulations for internal use for oral administration include pharmaceutically acceptable solutions, These liquid preparations include suspensions, emulsions, syrups, elixirs, etc. , one or more commonly used diluents (purified water, ethanol, mixed solutions of these, etc.) The liquid preparation dissolves, suspends or emulsifies the active ingredient. It may contain additives such as flavoring agents, sweeteners, flavoring agents, fragrances, preservatives, and buffering agents. Examples of topical dosage forms for use include ointments, gels, creams, ointments, Adhesive preparations, liniments, sprays, inhalants, sprays, aerosols, eye drops, nasal drops, etc. In addition, they can be sealed with biodegradable polymers and used in medical devices (surgical sutures, fracture treatments, etc.). These contain one or more active ingredients and are used in the conventional They are prepared by known methods or based on commonly used chemical formulas. In addition to the diluents used, sprays and inhalants contain stabilizers such as sodium bisulfite, and Buffers that can provide tonicity, such as sodium chloride, sodium citrate, and It may contain a tonicity agent such as enoic acid. Methods for producing the spray are described, for example, in U.S. Pat. Nos. 2,868,691 and 3,095,355. Solutions, suspensions, and emulsions that are dissolved or suspended in a solvent before use Injectables include solid injections and injections for parenteral administration. Injectables contain one or more active ingredients. These injections are administered by dissolving, suspending or emulsifying in a solvent. It can be injected into localized areas of the muscles, subcutaneous tissue, brain, joints, bones and other organs, or into a vascular catheter with a needle. It can be administered directly using a catheter, etc. Examples of solvents include distilled water for injection, Physiological saline, vegetable oil, propylene glycol, polyethylene glycol, ethanol, etc. In addition, such injections are safe and effective. Stabilizer, solubilizing agent (glutamic acid, aspartic acid, polysorbate 80 (registered trademark) These may contain additives such as suspending agents, emulsifying agents, buffers, preservatives, etc. These may be sterilized at the final stage. They are manufactured by aseptic processing or by aseptic manipulation. The agent may be prepared and dissolved in sterile or aseptic distilled water for injection or other solvents before use. It is possible. [Example]

[0059] The following examples are included to demonstrate preferred embodiments of the invention. The following techniques have been found by the inventors to work well in the practice of the present invention: and therefore can be considered to constitute a preferred mode for carrying out the invention. However, those skilled in the art will understand that, in view of the present disclosure, , and in specific embodiments disclosed and similar results can be obtained, are not intended to be construed as falling outside the spirit and scope of the present invention. It will be understood that many variations are possible without departing from the scope of the invention.

[0060] Example 1 Synergistic effect of fibroblasts and minocycline in suppressing inflammation Three concentrations of lipopolysaccharide (which activates TLR4, the receptor associated with CTE) were used. To mimic TLR4 activation, plate a 96-well plate with adherent monocytes at confluence. Fibroblasts alone, minocycline alone, and fibroblasts plus minocycline were added. The evaluation of inflammatory cytokines was tested: IL-1β (Fig. 1), TNF-α (Fig. 2), and IL-6 (Fig. 3). In Fig. 4, the fibroblasts induce T regulatory cells. It has been demonstrated that it enhances the potency of icrine.

[0061] References All patents and publications mentioned in the specification are incorporated by reference in their entirety to which this invention pertains. All patents and publications are incorporated by reference. All such references are incorporated herein by reference to the same extent as if specifically and individually indicated to be incorporated. will be incorporated into

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[0130] Although the present disclosure and its advantages have been described in detail, the scope of the present invention is defined by the appended claims. Various changes, substitutions, and alterations may be made herein without departing from the spirit and scope of the present design. It is understood that the scope of the present application is limited to the Any disclosures expressly incorporated herein by reference are limited to particular embodiments of the process, machine, manufacture, composition of matter, means, methods and steps. As those skilled in the art will readily understand from this disclosure, perform substantially the same function or achieve substantially the same result as the corresponding embodiment described in Any now existing or later developed process, machine, manufacture, composition of matter, or technique that accomplishes the Any stage, method, or step may be utilized in accordance with the present disclosure. The appended claims include within their scope such processes, machines, manufacture, compositions of matter, means, or methods. or steps.

Claims

1. A method of increasing the regenerative activity of a fibroblast population, comprising exposing said population to an NF-κB inhibitor.

2. A method according to claim 1, wherein the one or more NF-κB inhibitors suppress the ability of fibroblasts to produce one or more inflammatory cytokines.

3. A method according to claim 2, wherein the inflammatory cytokine is TNF-α.

4. A method according to claim 2 or 3, wherein the inflammatory cytokine is IL-1β.

5. A method according to any one of claims 2 to 4, wherein the inflammatory cytokine is IL-6.

6. A method according to any one of claims 1 to 5, wherein the one or more NF-κB inhibitors are administered together with an AMPK activator to an individual in need thereof.

7. The method described in claim 6, wherein the AMPK activator is metformin.

8. A method according to any one of claims 1 to 7, wherein the one or more NF-κB inhibitors are administered to an individual in need thereof before, simultaneously with, or after administration of the fibroblasts.

9. A method according to any one of claims 1 to 8, wherein the fibroblasts are obtained from an autologous, allogeneic or xenogeneic source for an individual in need.

10. The method according to any one of claims 1 to 9, wherein the one or more NF-κB inhibitors are selected from the group consisting of oxytetracycline, demeclocycline, methacycline, doxycycline, chlortetracycline, tetracycline, sansicycline, kerocaldin, 6-demethyl-6-deoxy-4-dedimethylaminotetracycline, tetracyclino-pyrazole, 7-chloro-4-dedimethylaminotetracycline, 4-hydroxy-4-dedimethylaminotetracycline, 12 α-Deoxy-4-dedimethylaminotetracycline, 5-hydroxy-6α-deoxy-4-dedimethylaminotetracycline, 4-dedimethylamino-12α-deoxyanhydrotetracycline, 7-dimethylamino-6-demethyl-6-deoxy-4-dedimethylaminotetracycline, tetracyclinonitrile, 4-oxo-4-dedimethylaminotetracycline 4,6-hemiketal, 4-oxo-11αC1-4-dedimethylaminotetracycline-4,6- Hemiketal, 5α,6-anhydro-4-hydrazono-4-dedimethylaminotetracycline, 4-hydroxyimino-4-dedimethylaminotetracyclines, 4-hydroxyimino-4-dedimethylamino-5α,6-anhydrotetracyclines, 4-amino-4-dedimethylamino-5α,6-anhydrotetracycline, 4-methylamino-4-dedimethylaminotetracycline, 4-hydrazono-11α-chloro-6-deoxy-6-demethyl-6-methylene-4 -dedimethylaminotetracycline, tetracycline quaternary ammonium compounds, anhydrotetracycline betaines, 4-hydroxy-6-methylpretetramids, 4-ketotetracyclines, 5-ketotetracyclines, 5α,11α-dehydrotetracyclines, 11αC1-6,12-hemiketal tetracyclines, 11αC1-6-methylenetetracyclines, 6,13-diol tetracyclines, 6-benzylthiomethylenetetracyclines, 7,11α-dichloro-6-fluoromethyl-6-deoxytetracyclines, 6-fluoro(α)-6-demethyl-6-deoxytetracyclines, 6-fluoro(β)-6-demethyl-6-deoxytetracyclines, 6-α-acetoxy-6-demethyltetracyclines, 6-β-acetoxy-6-demethyltetracyclines, 7,13-epithiotetracyclines, oxytetracyclines, pyrazolotetracyclines, 11α halogenated tetracyclines, 12α formyl and other tetracycline esters, 5,12α ester tetracyclines, 10,12α-diester tetracyclines The method is selected from lacyclines, isotetracycline, 12α-deoxyanhydrotetracyclines, 6-demethyl-12α-deoxy-7-chloroanhydrotetracyclines, B-nortetracyclines, 7-methoxy-6-demethyl-6-deoxytetracyclines, 6-demethyl-6-deoxy-5α-epitetracyclines, 8-hydroxy-6-demethyl-6-deoxytetracyclines, monardene, chromocycline, 5α-methyl-6-demethyl-6-deoxytetracyclines, 6-oxatetracyclines, 6-thiatetracyclines, and combinations thereof.

11. A method as described in claim 1, wherein the fibroblast population and the one or more inhibitors are provided to an individual in need thereof, optionally together with one or more additional agents that enhance the regenerative activity of the fibroblasts.

12. The method according to claim 11, wherein the additional agent is selected from the group consisting of a compound that removes protein accumulation (geldanamycin, etc.), an anti-inflammatory agent (glucocorticoid, nonsteroidal anti-inflammatory drug (ibuprofen, aspirin, etc.), omega-3 fatty acid (EPA, DHA, etc.), dexanabionol, etc.), a compound that increases energy available to cells (creatine, creatine phosphate, dichloroacetic acid, nicotinamide, riboflavin, carnitine, etc.), an antioxidant (plant extract (ginkgo biloba, etc.), coenzyme Q10, vitamin E (α-tocopherol), vitamin C (ascophyllum), etc.), and the like. inducing atherosclerosis; and / or rheumatoid arthritis. In some embodiments, the therapeutic agent is selected from the group consisting of steroids (such as steroids), anti-inflammatory drugs ...

13. A method according to any one of claims 1 to 12, wherein the fibroblasts are plastic-adherent.

14. A method according to any one of claims 1 to 13, wherein the fibroblasts express CD105.

15. A method according to any one of claims 1 to 14, wherein the fibroblasts express CD73.

16. A method for treating or preventing a neuropathy in an individual, the method comprising the step of administering to an individual having or at risk of neuropathy an effective amount of fibroblasts and one or more NF-κB inhibitors.

17. The method according to claim 16, wherein the one or more NF-κB inhibitors are selected from the group described in claim 10.

18. A method according to claim 16 or 17, wherein the fibroblasts are administered to an individual before, simultaneously with or after administration of the one or more inhibitors.

19. A method according to any one of claims 16 to 18, wherein the individual is a professional or recreational athlete or an individual engaged in an occupation involving a risk of head trauma.

20. A method according to any one of claims 16 to 19, wherein the individual is further administered one or more agents selected from the group described in claim 12.

21. A method according to any one of claims 16 to 20, wherein the neurological disorder is a central nervous system injury, a chronic injury, an acute injury, or a disease.

22. A method as described in claim 21, wherein the chronic injury is chronic traumatic encephalopathy.

23. A method according to any one of claims 16 to 22, wherein the neurological disorder is Alzheimer's disease (AD), age-related memory impairment, mild cognitive impairment, vascular dementia, acute spinal cord injury, amyotrophic lateral sclerosis (ALS), ataxia, Bell's palsy, brain tumor, cerebral aneurysm, epilepsy, convulsion, Guillain-Barré syndrome, meningitis, multiple sclerosis, muscular dystrophy, Parkinson's disease, migraine, stroke, encephalitis, myasthenia gravis, or a combination thereof.

24. A method for suppressing inflammation in an individual, comprising providing to said individual a population of fibroblasts and a therapeutically effective amount of one or more NF-κB inhibitors.

25. The method according to claim 24, wherein the one or more NF-κB inhibitors are selected from the group described in claim 10.

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