Smallpox vaccine and stem cells for treating disease

Abstract

Described herein are methods and compositions for treating an inflammatory disease or an infectious disease in a subject in need thereof by administering a poxvirus and stem cells to the subject, wherein the disease is not a cancer. For example, the disease can be a chronic inflammatory disease (e.g., an autoimmune disease).

Description

Background Technology

[0001] Inflammatory diseases, such as autoimmune diseases, are caused by chronic inflammation in the subjects. These diseases can lead to symptoms ranging from mild discomfort to severe reactions, and even death.

[0002] Infectious diseases are caused by organisms such as bacteria, viruses, fungi, or parasites. While infections are typically treated with antibiotics, antiviral drugs, antifungal drugs, antiprotozoal drugs, and antihelmintic drugs, pathogens are becoming increasingly resistant to these drugs. Other pathogens currently have no known treatments.

[0003] New methods are needed to treat these diseases. Summary of the Invention

[0004] This article describes methods and compositions for treating a disease in subjects by administering poxvirus and stem cells to subjects in need, wherein the disease is not cancer.

[0005] In one aspect, a method for treating a chronic inflammatory disease in a subject is provided. The method includes administering a poxvirus to the subject, wherein the disease is not cancer. In an embodiment, the poxvirus is administered in a therapeutically effective amount, for example, an amount sufficient to treat a chronic inflammatory disease.

[0006] In one aspect, a method is provided for treating an infectious disease and / or its symptoms in a subject. The method includes administering a poxvirus to the subject. In an embodiment, the poxvirus is administered in a therapeutically effective amount, for example, an amount sufficient to treat the infectious disease and / or its symptoms. In an embodiment, the disease is not cancer.

[0007] In one aspect, a method is provided for preventing an infectious disease and / or its symptoms in a subject. The method includes administering a poxvirus to the subject. In an embodiment, the poxvirus is administered in a therapeutically effective amount, for example, an amount sufficient to prevent the infectious disease and / or its symptoms. In an embodiment, the disease is not cancer.

[0008] In one aspect, a method for preventing a cytokine storm in a subject is provided. The method includes administering a poxvirus to the subject. In an embodiment, the poxvirus is administered in a therapeutically effective amount, for example, an amount sufficient to prevent a cytokine storm. In an embodiment, the cytokine storm is caused by an infection / infectious disease.

[0009] In one aspect, a method for treating a disease characterized by chronic inflammation is provided. The method includes administering a poxvirus and stem cells to a subject, wherein the disease is not cancer. In one embodiment, the poxvirus is administered in a therapeutically effective amount, e.g., an amount sufficient to treat the disease. In another embodiment, the stem cells are administered in a therapeutically effective amount, e.g., an amount sufficient to treat the disease.

[0010] In the implementation scheme, poxvirus and stem cells are in the same (single) composition.

[0011] In one aspect, a method is provided for converting chronic inflammation into acute inflammation in a subject with this need. The method includes administering a poxvirus to the subject, wherein the condition is not cancer. In an embodiment, the poxvirus is administered in a therapeutically effective amount, for example, an amount sufficient to convert chronic inflammation into acute inflammation. In an embodiment, the method also includes treating the acute inflammation. In an embodiment, treating the acute inflammation includes administering a known treatment for acute inflammation to the subject.

[0012] In one aspect, a composition comprising stem cells and optionally a poxvirus is provided, wherein the poxvirus comprises a recombinant polynucleotide encoding a therapeutic molecule. In one embodiment, the therapeutic molecule treats a chronic inflammatory disease. In another embodiment, the therapeutic molecule is an anti-inflammatory molecule.

[0013] In the implementation plan, the disease is a chronic inflammatory disease. In the implementation plan, the chronic inflammatory disease is an autoimmune disease. In the implementation plan, the chronic inflammatory disease is asthma, chronic peptic ulcer, tuberculosis, arthritis, periodontitis, ulcerative colitis, Crohn's disease, sinusitis, active hepatitis, atherosclerosis, dermatitis, inflammatory bowel disease (IBS), systemic lupus erythematosus, fibromyalgia, type 1 diabetes, psoriasis, multiple sclerosis, Addison's disease, Grave's disease, Sjögren's syndrome (…). Syndrome, Hashimoto's thyroiditis, myasthenia gravis, vasculitis, pernicious anemia, or celiac disease.

[0014] In one implementation, the inflammatory disease is transplant rejection, Dupytren's contracture, peyronies, periodontitis, endometriosis, hepatitis, glomerulonephritis, atherosclerosis, cardiovascular disease, arthritis (e.g., osteoarthritis, rheumatoid arthritis, or psoriatic arthritis), inflammatory encephalopathy (including post-stroke, encephalitis), atherosclerosis, traumatic injury, infection, and / or shock. In one implementation, the inflammatory disease is chronic obstructive pulmonary disease (COPD), such as emphysema, chronic bronchitis, or refractory (irreversible) asthma.

[0015] In the implementation plan, inflammatory diseases are enterocutaneous fistulas, chronic radiation injuries (which result in inflammatory tissue defects, such as radiation cystitis or radiation enteritis), duodenal ulcers, or chronic inflammatory diseases of the central nervous system, such as post-stroke neuroinflammation, schizophrenia, autism, addiction, chronic traumatic encephalopathy, or vaccine-induced neurotoxicity.

[0016] In one implementation scheme, the autoimmune disease is myasthenia gravis (MG), Hashimoto's thyroiditis, vasculitis, Gray's disease, psoriasis, chronic inflammatory demyelinating polyneuropathy (CIDP), Guillain-Barré syndrome, type 1 diabetes, lupus, multiple sclerosis, rheumatoid arthritis, Addison's disease, Sjögren's syndrome, celiac disease, myositis, ankylosing spondylitis, or scleroderma.

[0017] In one embodiment, the infectious disease is caused by bacteria, a virus, or a fungus. In one embodiment, the infectious disease is caused by a virus. In one embodiment, the virus is rhinovirus, coronavirus, influenza, or respiratory syncytial virus. In one embodiment, the coronavirus is severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2).

[0018] In one implementation, an infectious disease causes or is capable of causing a cytokine storm in the subject. In another implementation, an inflammatory disease causes or is capable of causing a cytokine storm in the subject.

[0019] In one implementation, the stem cells comprise recombinant polynucleotides. In another implementation, the recombinant polynucleotides encode a therapeutic molecule. In yet another implementation, the poxvirus comprises recombinant polynucleotides. In yet another implementation, the recombinant polynucleotides encode a therapeutic molecule.

[0020] In one embodiment, the therapeutic molecule treats a disease. In another embodiment, the therapeutic molecule is a cytokine, a therapeutic antibody, a therapeutic fusion protein, RNA, a peptide, or a polypeptide. In another embodiment, the cytokine is an anti-inflammatory cytokine. In yet another embodiment, the cytokine is selected from interleukin (IL)-1 receptor antagonists, IL-4, IL-6, IL-10, IL-11, IL-13, IFN-α, and transforming growth factor-β. In the implementation scheme, the therapeutic molecule is selected from Orencia, Humira, Kinetet, Cimzia, Enbrel, Simponi, Remicade, Taltz, Tysabri, Rituxan, Cosentyx, Actemra, Stelara, Entyvio, Simulect, Zinbryta, and Orthoclone OKT3.

[0021] In this embodiment, the therapeutic molecule improves the treatment of a disease. For example, the therapeutic molecule can be a receptor that promotes the uptake of a therapeutic agent by cells expressing the therapeutic molecule. In another example, the therapeutic molecule can be an antigen recognized by the therapeutic agent. In yet another example, the therapeutic molecule can be an enzyme used by cells to produce a therapeutic agent (e.g., a steroid). In this embodiment, the therapeutic agent is a pharmaceutical agent that treats a disease.

[0022] In one embodiment, the method further includes administering a therapeutic agent to a subject. In another embodiment, the therapeutic agent is in the same composition as the poxvirus and stem cells. In yet another embodiment, the therapeutic agent is administered separately from the poxvirus and stem cells. In one embodiment, the therapeutic agent is a drug for treating a disease. In another embodiment, the therapeutic agent is selected from Orencia, Humira, Kinetetin, Cimzia, Enbrel, Simponi, Remicade, Taltz, Tysabri, Rituxan, Cosentyx, Actemra, Stelara, Entyvio, Simulect, Zinbryta, and Orthoclone OKT3.

[0023] In one implementation, poxvirus and optional stem cells are administered to the subject via intravenous injection, intraperitoneal injection, intrathecal injection, intraventricular injection, intra-articular injection, intraventricular injection, intrapleural injection, intraparencymal injection, or intraocular injection. In another implementation, poxvirus and optional stem cells are administered directly to the area affected by the disease. In yet another implementation, poxvirus and optional stem cells are administered via guided delivery, such as MRI-guided delivery.

[0024] In this implementation plan, the stem cells are autologous to the subject. In this implementation plan, the stem cells are allogeneic to the subject. In this implementation plan, the subject is human. In this implementation plan, the subject is a non-human animal. In this implementation plan, the subject is a domesticated animal. In this implementation plan, the subject is a companion animal.

[0025] In the implementation scheme, the poxvirus is a vaccinia virus. In the implementation scheme, the vaccinia virus is selected from Dryvax, ACAM1000, ACAM2000, Lister, EM63, LIVP, Tian Tan, Copenhagen, Western Reserve, Modified Vaccinia Ankara (MVA), New York City Board of Health, Dairen, Ikeda, LC16M8, Western Reserve Copenhagen, Tashkent, Tian Tan, Wyeth, IHD-J and IHD-W, Brighton, Dairen I, and Connaught strains. In the implementation scheme, the vaccinia virus is ACAM1000 or ACAM2000. In the implementation scheme, the vaccinia virus is a New York City Board of Health strain. In the implementation scheme, the poxvirus is an attenuated virus.

[0026] In the implementation plan, the stem cells are selected from adult stem cells, embryonic stem cells, fetal stem cells, mesenchymal stem cells, neural stem cells, totipotent stem cells, pluripotent stem cells, multipotent stem cells, oligopotent stem cells, unipotent stem cells, adipose stromal cells, endothelial stem cells, induced pluripotent stem cells, bone marrow stem cells, umbilical cord blood stem cells, adult peripheral blood stem cells, myoblast stem cells, juvenile small stem cells, skin fibroblast stem cells, and combinations thereof. In the implementation plan, the stem cells are derived from the subject to be treated with the composition.

[0027] In this embodiment, the stem cell is a modified stem cell. In this embodiment, the modified stem cell expresses (is modified to express) a heterologous protein. In this embodiment, the heterologous protein is a therapeutic molecule, a receptor that promotes the uptake of a therapeutic agent, an antigen recognized by the therapeutic agent, or an enzyme involved in the production of the therapeutic agent. In this embodiment, the therapeutic agent is a drug for treating a disease. Invention Details

[0029] After reading this description, it will become apparent to those skilled in the art how the invention can be practiced in various alternative embodiments and applications. However, not all various embodiments of the invention will be described herein. It should be understood that the embodiments presented herein are merely illustrative and not limiting. Therefore, this detailed description of various alternative embodiments should not be construed as limiting the scope or breadth of the invention as described below.

[0030] Before disclosing and describing the present invention, it should be understood that the aspects described below are not limited to specific compositions, methods of preparing such compositions, or uses thereof, as these can certainly vary. It should also be understood that the terminology used herein is for the purpose of describing specific aspects only and is not intended to be limiting.

[0031] The detailed description of the invention has been divided into various sections for the convenience of the reader, and the disclosures in any section can be combined with the disclosures in another section. Titles or subtitles may be used in the specification for the reader's convenience, but this is not intended to affect the scope of the invention.

[0032] I. Definition

[0033] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. In this specification and the following claims, numerous terms will be referenced and defined as having the following meanings:

[0034] The terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the invention. Unless the context clearly indicates otherwise, the singular forms “an,” “a,” and “the” as used herein are intended to include the plural forms as well.

[0035] "Optional" or "optionally" means that the event or situation described below may or may not occur, and the description includes instances of the event or situation occurring and instances of it not occurring.

[0036] When the term "about" is used before numerical designations (e.g., temperature, time, amount, concentration, and such names, including ranges), it indicates an approximate value that can vary (+) or (-) 10%, 5%, 1%, or any subrange or subvalue between these values. Preferably, when used in relation to dosage, the term "about" means that the dosage can vary by + / - 10%.

[0037] The terms "comprising" or "including" mean that a composition and method include the listed elements but do not exclude other elements. When used to define compositions and methods, "consisting substantially of..." should mean excluding other elements that are of any significance to the composition for the stated purpose. Therefore, a composition consisting substantially of the elements defined herein does not exclude other materials or steps that do not substantially affect the essential and novel features of the claimed invention. "Constitutes..." should mean excluding other ingredients and substantial method steps beyond trace amounts. Embodiments defined by each of these transitional terms are within the scope of this invention.

[0038] The terms "disease" or "condition" refer to the survival status or health condition of a patient or subject who can be treated with the compounds or methods provided herein. The disease can be an autoimmune disease. The disease can be an inflammatory disease. The disease can be an infectious disease.

[0039] As used herein, the term "inflammatory disease" refers to a disease or condition characterized by abnormal inflammation (e.g., elevated levels of inflammation compared to a control group, such as a healthy person without the disease). The term "chronic inflammatory disease" refers to a persistent or recurrent inflammatory disease. Examples of chronic inflammatory diseases include autoimmune diseases, arthritis, rheumatoid arthritis, psoriatic arthritis, juvenile idiopathic arthritis, multiple sclerosis, systemic lupus erythematosus (SLE), myasthenia gravis, juvenile diabetes mellitus, type 1 diabetes mellitus, Guillain-Barré syndrome, Hashimoto's encephalitis, Hashimoto's thyroiditis, ankylosing spondylitis, psoriasis, Sjögren's syndrome, vasculitis, glomerulonephritis, autoimmune thyroiditis, Behcet's disease, Crohn's disease, ulcerative colitis, bullous pemphigoid, sarcoidosis, ichthyosis, and Graves' ophthalmopathy. ophthalmopathy, inflammatory bowel disease, Addison's disease, vitiligo, asthma, allergic asthma, acne vulgaris, celiac disease, chronic prostatitis, inflammatory bowel disease, pelvic inflammatory disease, reperfusion injury, ischemia-reperfusion injury, stroke, sarcoidosis, transplant rejection, interstitial cystitis, atherosclerosis, scleroderma, and atopic dermatitis.

[0040] In the implementation plan, inflammatory diseases are enterocutaneous fistulas, chronic radiation injuries (which result in inflammatory tissue defects, such as radiation cystitis or radiation enteritis), duodenal ulcers, or chronic inflammatory diseases of the central nervous system, such as post-stroke neuroinflammation, schizophrenia, autism, addiction, chronic traumatic encephalopathy, or vaccine-induced neurotoxicity.

[0041] In one implementation scheme, inflammatory diseases include transplant rejection, Dupytren's contracture, Peronis disease, periodontitis, endometriosis, hepatitis, glomerulonephritis, atherosclerosis, cardiovascular disease, arthritis (e.g., osteoarthritis, rheumatoid arthritis, or psoriatic arthritis), inflammatory encephalopathy (including post-stroke, encephalitis), atherosclerosis, traumatic injury, infection, and / or shock. In another implementation scheme, an inflammatory disease is chronic obstructive pulmonary disease (COPD), such as emphysema, chronic bronchitis, or refractory (irreversible) asthma.

[0042] As used in this article, the term “autoimmune disease” refers to a disease or condition in which a subject’s immune system produces an abnormal immune response to a substance that would not normally elicit an immune response in a healthy subject. Examples of autoimmune diseases that can be treated with the compounds, pharmaceutical compositions, or methods described herein include acute disseminated encephalomyelitis (ADEM), acute necrotizing hemorrhagic leukoencephalitis, Addison's disease, agammaglobulinemia, alopecia areata, amyloidosis, ankylosing spondylitis, anti-GBM / anti-TBM nephritis, antiphospholipid syndrome (APS), autoimmune angioedema, autoimmune aplastic anemia, autoimmune autonomic dysfunction, autoimmune hepatitis, autoimmune hyperlipidemia, autoimmune immunodeficiency, autoimmune inner ear disease (AIED), autoimmune myocarditis, autoimmune oophoritis, autoimmune pancreatitis, autoimmune retinopathy, autoimmune thrombocytopenic purpura (ATP), autoimmune thyroid disease, autoimmune urticaria, axonal or neuronal neuropathy, Balo's disease, and Behcet's disease. Diseases including bullous pemphigoid, cardiomyopathy, Castleman's disease, celiac disease, Chagas disease, chronic fatigue syndrome, chronic inflammatory demyelinating polyneuropathy (CIDP), chronic relapsing multifocal osteomyelitis (CRMO), Churg-Strauss syndrome, cicatricial pemphigoid / benign mucosal pemphigoid, Crohn's disease, Cogans syndrome, cold agglutinin disease, congenital heart block, Coxsackievirus myocarditis, CREST disease, primary mixed cryoglobulinemia, demyelinating neuropathy, herpetic dermatitis, dermatomyositis, Dervex disease (neuromyelitis optica), discoid lupus, Dressler's syndrome, endometriosis, eosinophilic esophagitis, eosinophilic fasciitis, erythema nodosum, experimental allergic encephalomyelitis, and Evans syndrome. Fibromyalgia, fibrosomnitis, giant cell arteritis (temporal arteritis), giant cell myocarditis, glomerulonephritis, pulmonary hemorrhage-nephritis syndrome (Goodpasture's syndrome)Gram-positive syndrome, granulomatous polyangiitis (GPA) (formerly known as Wegener's granulomatosis), Graves' disease, Guillain-Barré syndrome, Hashimoto's encephalitis, Hashimoto's thyroiditis, hemolytic anemia, Henoch-Schonleinpurpura, herpes gestationis, hypogammaglobulinemia, idiopathic thrombocytopenic purpura (ITP), IgA nephropathy, IgG4-related sclerosis, immunomodulatory lipoprotein, inclusion body myositis, interstitial cystitis, juvenile arthritis, juvenile diabetes mellitus (type 1 diabetes), juvenile myositis, Kawasaki syndrome, Lambert-Eaton syndrome Syndrome, leukocytic granulomatosis, lichen planus, lichen sclerosus, woody conjunctivitis, linear IgA disease (LAD), lupus (SLE), Lyme disease, chronic diseases, Meniere's disease, microscopic polyangiitis, mixed connective tissue disease (MCTD), Mooren's ulcer, Mucha-Habermann disease, multiple sclerosis, myasthenia gravis, myositis, narcolepsy, neuromyelitis optica (Devi's disease), neutropenia, ocular cicatricial pemphigoid, optic neuritis, relapsing rheumatism, PANDAS (pediatric autoimmune neuropsychiatric disorder with streptococcal infection), paraneoplastic cerebellar degeneration, paroxysmal nocturnal hemoglobinuria (PNH), Parry-Romberg syndrome, Parsonnage-Turner syndrome. Symptoms of cilia flats (peripheral uveitis), pemphigus, peripheral neuropathy, perivenous encephalomyelitis, pernicious anemia, POEMS syndrome, polyarteritis nodosa, type I, II and III autoimmune polyglandular syndrome, polymyalgia rheumatica, polymyositis, post-myocardial infarction syndrome, post-pericardiotomy syndrome, progesterone dermatitis, primary biliary cirrhosis, primary sclerosing cholangitis, psoriasis, psoriatic arthritis, idiopathic pulmonary fibrosis, pyoderma gangrenosa, simple erythropoiesis, Raynaud's phenomenon, reactive arthritis, reflex sympathetic dystrophy, Reiter's syndrome, relapsing polychondritis, restless extremities syndrome, retroperitoneal fibrosis, rheumatic fever, rheumatoid arthritis, sarcoidosis, Schmidt's syndromesyndrome), scleritis, scleroderma, Sjögren's syndrome, sperm and testis autoimmunity, stiff-person syndrome, subacute bacterial endocarditis (SBE), Susac's syndrome, sympathetic ophthalmia, Takayasu'sarteritis, temporal arteritis / giant cell arteritis, thrombocytopenic purpura (TTP), Tolosa-Hunt syndrome, transverse myelitis, type 1 diabetes mellitus, ulcerative colitis, undifferentiated connective tissue disease (UCTD), uveitis, vasculitis, bullous dermatitis, vitiligo, or Wegener's granulomatosis (i.e., granulomatous polyangiitis (GPA)).

[0043] In the implementation plan, inflammatory diseases are enterocutaneous fistulas, chronic radiation injuries (which result in inflammatory tissue defects, such as radiation cystitis or radiation enteritis), duodenal ulcers, or chronic inflammatory diseases of the central nervous system, such as post-stroke neuroinflammation, schizophrenia, autism, addiction, chronic traumatic encephalopathy, or vaccine-induced neurotoxicity.

[0044] In one implementation scheme, the autoimmune disease is myasthenia gravis (MG), Hashimoto's thyroiditis, vasculitis, Gray's disease, psoriasis, chronic inflammatory demyelinating polyneuropathy (CIDP), Guillain-Barré syndrome, type 1 diabetes, lupus, multiple sclerosis, rheumatoid arthritis, Addison's disease, Sjögren's syndrome, celiac disease, myositis, ankylosing spondylitis, or scleroderma.

[0045] The term "treating" or "treatment" refers to any indicator of success in the treatment or improvement of an injury, disease, lesion, or condition, including any objective or subjective parameters such as reduction; relief; decrease in symptoms or make the injury, lesion, or condition more tolerable for the patient; slowing of the rate of deterioration or decline; making the endpoint of deterioration less debilitating; or improving the patient's physical or mental health. Treatment or improvement of symptoms can be based on objective or subjective parameters; including the results of physical examination, neuropsychiatric examination, and / or psychiatric evaluation. Variations of the term "treatment" and its verb form can include prevention of an injury, lesion, condition, or disease. In this implementation, treatment is prevention. In this implementation, treatment does not include prevention.

[0046] As used herein, “treating” or “treatment” (and as is well understood in the art) also broadly includes any means used to obtain a beneficial or desired outcome (including clinical outcomes) in relation to the condition of a subject. Beneficial or desired clinical outcomes may include, but are not limited to, alleviating or improving one or more symptoms or conditions, reducing the extent of the disease, stabilizing (i.e., not worsening) the state of the disease, preventing the spread or diffusion of the disease, delaying or slowing the progression of the disease, improving or alleviating the state of the disease, reducing recurrence of the disease, and remission, whether partial or complete, detectable or undetectable. In other words, as used herein, “treatment” includes any cure, improvement, or prevention of the disease. Treatment may prevent the onset of the disease; inhibit the spread of the disease; alleviate the symptoms of the disease (e.g., eye pain, seeing halos around light sources, red eyes, very high intraocular pressure), completely or partially eliminate the root cause of the disease, shorten the duration of the disease, or a combination of these.

[0047] As used herein, “treating” and “treatment” include prophylactic treatment. Treatment methods include administering a therapeutically effective amount of an active agent to a subject. Administration may consist of a single administration or may include a series of administrations. The duration of treatment depends on various factors, such as the severity of the condition, the patient's age, the concentration of the active agent, the activity of the composition used in the treatment, or a combination thereof. It should also be understood that the effective dose of the agent used for treatment or prevention may be increased or decreased during a particular treatment or prevention regimen. Changes in dose can occur and become apparent using standard diagnostic assays known in the art. In some cases, prolonged administration may be required. For example, the composition may be administered to the subject in a dose and duration sufficient to treat the patient. In embodiments, treatment is not prophylactic treatment.

[0048] The term "prevention" refers to a reduction in the incidence of disease symptoms in patients. As mentioned above, prevention can be complete (no detectable symptoms) or partial, such that fewer symptoms are observed than would be present without treatment.

[0049] "Patient," "subject," or "subject in need" refers to a living organism that suffers from or is susceptible to a disease or condition that can be treated by administration of the pharmaceutical composition provided herein. Non-limiting examples include humans, other mammals, bovine animals, rats, mice, dogs, monkeys, goats, sheep, cows, deer, and other non-mammalian animals. In some embodiments, the patient is a human. In some embodiments, the person is a pediatric patient. In some embodiments, the patient is a domesticated animal (e.g., goats, sheep, cows, horses, etc.). In some embodiments, the patient is a companion animal, including but not limited to canines, felines, rodents (mice, rats, gerbils, hamsters, guinea pigs, chinchillas, etc.), rabbits, ferrets, etc.

[0050] "Effective amount" means an amount sufficient to enable a compound to achieve its intended purpose (e.g., to achieve the effect of the compound, to treat a disease, or to alleviate one or more symptoms of a disease or condition) relative to the absence of the compound. An example of "effective amount" is an amount sufficient to help treat, prevent, or alleviate one or more symptoms of a disease, which may also be called "therapeutic effective amount". "Amelioration" of one or more symptoms (and its grammatical equivalent) means a reduction in the severity or frequency of the symptoms, or the elimination of the symptoms. The exact amount will depend on the purpose of treatment and can be determined by a person skilled in the art using known techniques (see, for example, Lieberman, Pharmaceutical Dosage Forms (Vols. 1–3, 1992); Lloyd, The Art, Science and Technology of Pharmaceutical Compounding (1999); Pickar, Dosage Calculations (1999); and Remington: The Science and Practice of Pharmacy, 20th Edition, 2003, Gennaro, ed., Lippincott, Williams & Wilkins).

[0051] As is well known in the art, therapeutically effective doses for humans can also be determined from animal models. For example, human doses can be formulated to achieve the levels found to be effective in animals. As described herein, human doses can be adjusted by monitoring efficacy and adjusting the dose upward or downward. Adjusting the dose according to the methods described herein and other methods to achieve maximum efficacy in humans is entirely within the capabilities of a person skilled in the art.

[0052] As used herein, the term "therapeutic effective dose" refers to the amount of a therapeutic agent sufficient to improve the condition as described above. For example, for a given parameter, a therapeutic effective dose will show an increase or decrease of at least 5%, 10%, 15%, 20%, 25%, 40%, 50%, 60%, 75%, 80%, 90%, or at least 100%. Therapeutic efficacy can also be expressed as an increase or decrease in "multiples." For example, the effect of a therapeutic effective dose can be at least 1.2 times, 1.5 times, 2 times, 5 times, or more than that of the control.

[0053] Dosage can vary depending on the patient's needs and the composition used. In the context of this disclosure, the dose administered to the patient should be sufficient to achieve a beneficial therapeutic response in the patient over time. The magnitude of the dose will also depend on the presence, nature, and extent of any adverse side effects. The appropriate dose for a particular situation is determined within the skill level of the practitioner. Generally, a dose less than the optimal dose of the composition is used at the start of treatment. Thereafter, the dose is increased in small increments until the optimal effect for the situation is achieved. Dosage and intervals can be adjusted individually to provide a level of administered composition effective for the specific clinical indication for which treatment is being received. This will provide a treatment regimen commensurate with the severity of the individual's disease state.

[0054] As used herein, the term "administration" means oral administration to a subject, as a suppository, local contact administration, intravenous administration, parenteral administration, intraperitoneal administration, intramuscular administration, intralesional administration, intrathecal administration, intrathecal administration, intraventricular administration, intrapleural administration, intraparenchymal administration, intranasal administration, or subcutaneous administration, or administration via implantation of a sustained-release device, such as a micro-osmotic pump. Administration can be made via any route, including parenteral and transmucosal administration (e.g., buccal, sublingual, palatal, gingival, nasal, vaginal, rectal, or percutaneous administration). Parenteral administration includes, for example, intravenous, intramuscular, microarterial, intradermal, subcutaneous, intraperitoneal, intraventricular, and intracranial administration. Other delivery methods include, but are not limited to, the use of liposomal formulations, intravenous infusion, etc. Administration also includes direct administration, such as direct administration to the site of inflammation. Direct administration can be via guided delivery, such as magnetic resonance imaging (MRI) guided delivery. In embodiments, administration does not include the administration of any active agent other than the described active agent.

[0055] "Co-administration" means administering the compositions described herein concurrently with, just before, or immediately after the administration of one or more other therapies. The compositions provided herein can be administered alone or co-administered to a patient. Co-administration means administering the compositions simultaneously or sequentially, either separately or in combination (of more than one composition). Therefore, these formulations may also be combined with other active substances when needed.

[0056] A stem cell is a cell characterized by its ability to self-renew through mitotic division and its potential to differentiate into tissues or organs. In mammalian stem cells, embryonic stem cells (ES cells) and adult stem cells (e.g., HSCs, IPSCs) can be distinguished. Embryonic stem cells reside in the blastocyst and produce embryonic tissues, while adult stem cells reside in adult tissues and are used for tissue regeneration and repair.

[0057] The term "infection" or "infectious disease" refers to a disease or condition caused by an organism such as bacteria, viruses, fungi, or any other pathogenic microorganism. In an embodiment, an infectious disease is caused by a pathogenic bacterium. Pathogenic bacteria are bacteria that cause disease (e.g., in humans). In an embodiment, an infectious disease is a bacterial-associated disease (e.g., tuberculosis caused by Mycobacterium tuberculosis). Non-restrictive bacterial-associated diseases include pneumonia, which may be caused by bacteria such as Streptococcus and Pseudomonas; or foodborne illnesses, which may be caused by bacteria such as Shigella, Campylobacter, and Salmonella. Bacterial-associated diseases also include tetanus, typhoid fever, diphtheria, syphilis, and leprosy. In the implementation plan, the infectious disease is bacterial vaginosis (i.e., bacterial overgrowth that alters the vaginal microbiota, displacing lactobacilli that maintain a healthy vaginal microbiota) (e.g., yeast infection or trichomoniasis); bacterial meningitis (i.e., bacterial inflammation of the meninges); bacterial pneumonia (i.e., bacterial infection of the lungs); urinary tract infection; bacterial gastroenteritis; or bacterial skin infection (e.g., impetigo or cellulitis). In one implementation, the infectious disease is an infection with Campylobacter jejuni, Enterococcus faecalis, Haemophilus influenzae, Helicobacter pylori, Klebsiella pneumoniae, Legionella pneumophila, Neisseria gonorrhoeae, Neisseria meningitides, Staphylococcus aureus, Streptococcus pneumoniae, or Vibrio cholerae. In another implementation, the infection is caused by spirochetes or Treponema pallidum, for example, an infection associated with Lyme disease.

[0058] In its usual and conventional sense, the term "immune response" and similar terms refer to an organism's response to protect itself from disease. This response can be carried out by either the innate or adaptive immune system, as is well known in the art.

[0059] As used herein, the term "vaccine" refers to any type of biological agent that contributes to or induces an active immune response against a specific disease or pathogen. Such biological agents may include, but are not limited to, antigens derived from pathogenic mediators or portions thereof. Such biological agents may also be in the form of attenuated live preparations, including live, weakened, or modified pathogenic mediators or pathogens; or may be present in the form of inactivated or inactivated pathogenic mediators or pathogens. Alternative forms of such biological agents include, but are not limited to, subunit forms, toxoid forms, conjugate forms, DNA and recombinant vector forms, or any suitable forms that may be developed or utilized in the future to induce an active immune response against them.

[0060] It should be noted that in some implementations, although the term "vaccine" is used herein, the vaccine does not need to provide significant immunity against smallpox (or any other pathogen) as long as it is effective against the disease described herein. For example, a vaccine can be any immunogenic or infectious composition that treats a disease. In some cases, the term is used to identify certain materials or compositions, rather than necessarily the ability of the material or composition to provide, for example, immunity against smallpox. The virus can be derived from any strain of virus, including, for example, one or more strains listed below and elsewhere herein, including those that are not part of an approved or intended vaccine.

[0061] As used herein, “virus” refers to any one of a large group of entities known as viruses. Typically, viruses consist of a protein coat surrounding a core of genetic material, RNA or DNA, but lack a semi-permeable membrane and can only grow and reproduce within living cells. Viruses used in the methods provided herein include, but are not limited to, poxviruses, adenoviruses, herpes simplex virus, Newcastle disease virus, vesicular stomatitis virus, mumps virus, influenza virus, measles virus, reovirus, human immunodeficiency virus (HIV), hantavirus, myxoma virus, cytomegalovirus (CMV), lentiviruses, and any plant or insect virus.

[0062] As used in this article, "heterologous nucleic acid" refers to nucleic acid, DNA, or RNA that has been introduced into a virus or cell (or the ancestor of a cell). Such heterologous nucleic acids may contain gene sequences and operable regulatory elements. For example, heterologous nucleic acids may contain selection marker genes, suicide genes, or genes that express useful protein products that are not expressed endogenously or are expressed at low levels endogenously.

[0063] As used herein, when referring to the application of poxvirus and cells, the term "concurrently" means that they are applied to each other within 48 hours. In some embodiments, poxvirus and cells are applied to each other within 36 hours, 24 hours, 12 hours, 10 hours, 8 hours, 6 hours, 4 hours, 2 hours, or 1 hour.

[0064] As used in this article, the terms "autologous," "autologous cell," or "autologous transplantation" indicate that the cell donor and recipient are the same individual. Conversely, the terms "allogeneic," "allogeneic cell," or "allogeneic transplantation" indicate that the cell donor and recipient are different individuals of the same species.

[0065] As used in this article, "adipose tissue" refers to body adipose tissue, a loose connective tissue primarily composed of adipocytes. In addition to adipocytes, adipose tissue also contains the cellular stromal vascular fraction (SVF), including preadipocytes, fibroblasts, vascular endothelial cells, various immune cells, and regenerative stem cells.

[0066] As used in this article, "adipose-derived stem cells" or "adult stem cells derived from adipose tissue (ADASC)" refers to stem cells that can be isolated from adipose tissue; these are pluripotent stem cells capable of differentiating into various cell types. Adipose-derived stem cells have been found to be comparable to, and even superior to, bone marrow stem cells in terms of cell differentiation potential, angiogenesis, and anti-inflammatory effects.

[0067] "Pharmaceutically acceptable excipients" and "pharmaceutically acceptable carriers" refer to substances that facilitate administration of the active agent to a subject and / or absorption of the active agent by the subject, and can be included in the compositions of this disclosure without causing significant adverse toxicological effects on the patient. Non-limiting examples of pharmaceutically acceptable excipients include water, NaCl, physiological saline solutions, lactated Ringer's solution, normal sucrose, normal glucose, binders, fillers, disintegrants, lubricants, coatings, sweeteners, flavoring agents, saline solutions (e.g., Ringer's solution), alcohols, oils, gelatin, carbohydrates such as lactose, amylose or starch, fatty acid esters, hydroxymethyl cellulose, polyvinylpyrrolidone, and colorants, etc. Such preparations can be sterilized, and if necessary, can be mixed with adjuvants such as lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure, buffers, colorants, and / or aromatic substances, etc., which do not react adversely with the compounds of this disclosure. Those skilled in the art will recognize that other pharmaceutical excipients are also useful in this disclosure.

[0068] II. Poxvirus

[0069] Smallpox virus is the cause of smallpox. Unlike the smallpox virus, vaccinia virus does not cause systemic disease in immunocompetent individuals under normal circumstances, and therefore it has been used as a live vaccine for immunization against smallpox. Due to the successful use of vaccinia virus vaccination worldwide, smallpox has been eradicated as a natural disease. Routine smallpox vaccination has been discontinued for many years, except for those at high risk of poxvirus infection (e.g., laboratory workers). Although the United States discontinued routine childhood immunization against smallpox in 1972, the use of smallpox vaccine for pediatric application is generally considered safe.

[0070] Attenuated strains derived from pathogenic viruses can be used to produce live vaccines. Non-limiting examples of viral strains already used as smallpox vaccines include, but are not limited to, Lister (also known as Elstree), the New York City Department of Health (“NYCBOH strain”), Dairen, Ikeda, LC16M8, Western Reserve (WR), Copenhagen, Tashkent, Tian Tan, Wyeth, IHD-J and IHD-W, Brighton, Ankara, MVA, Dairen I, LIPV, LC16MO, LIVP, WR 65-16, EM63, and Connaught strains. In some embodiments, the smallpox vaccine disclosed herein is an attenuated vaccinia virus New York City Department of Health (NYCBOH) strain. In some embodiments, the vaccinia virus NYCBOH strain may be ATCC VR-118 or CJ-MVB-SPX.

[0071] In some implementation schemes, the smallpox vaccine is non-attenuated.

[0072] In some implementations, the smallpox vaccine is selected from Dryvax, ACAM1000, ACAM2000, Lister, EM63, LIVP, Tian Tan, Copenhagen, Western Reserve, or Modified Vaccinia Ankara (MVA). In some implementations, the smallpox vaccine does not lack any genes present in one or more of these strains.

[0073] In some implementations, the smallpox vaccine is a replicating virus. In other implementations, the smallpox vaccine is a replication-defective virus.

[0074] III. Stem Cells

[0075] Following systemic administration, the virus undergoes significant elimination and / or neutralization. Stem cells serve as a medium to shield the disclosed smallpox vaccine from the influence of humoral and cellular immune components in the bloodstream. See, for example, U.S. Patent No. 10,105,436, the entire contents of which are incorporated herein by reference.

[0076] Accordingly, stem cells can be used as a medium for delivering poxviruses in vivo to disease sites in a subject. In some embodiments, the poxvirus is mixed with stem cells to prevent the immune system from clearing the virus before it reaches the desired site. Therefore, in some embodiments, a method for treating a disease in a subject is disclosed herein, comprising simultaneously administering poxvirus and stem cells to the subject. In some embodiments, the poxvirus does not contain heterologous nucleic acid. In some embodiments, the method further includes simultaneously administering a genetically engineered virus to the subject. Some embodiments relate to methods for preparing a stem cell and poxvirus composition prior to administration.

[0077] In some implementations, the mediator stem cells are autologous stem cells. In other cases, they are non-autologous or allogeneic.

[0078] In some embodiments, the stem cells are selected from adult stem cells, embryonic stem cells, fetal stem cells, mesenchymal stem cells, neural stem cells, totipotent stem cells, pluripotent stem cells, multipotent stem cells, oligopotent stem cells, unipotent stem cells, adipose stromal cells, endothelial stem cells, induced pluripotent stem cells, bone marrow stem cells, umbilical cord blood stem cells, adult peripheral blood stem cells, myoblast stem cells, juvenile small stem cells, skin fibroblast stem cells, and combinations thereof. In some embodiments, the modified stem cells are umbilical cord-derived mesenchymal-like cells. In some embodiments, the umbilical cord-derived mesenchymal-like cells are IMMSTEM cells. TM Cells. In some embodiments, stem cells are adipose-derived stromal cells. One or more of the cells listed above may be specifically excluded from certain compositions and methods.

[0079] In some embodiments, the stem cells are modified. Particularly in some embodiments, the modified stem cells are adult stem cells (ASCs). In some embodiments, the modified stem cells are transformed with a viral vector. In some embodiments, the modified stem cells are transformed with a lentivirus or retrovirus. In some embodiments, the modified stem cells are transformed with a recombinant virus. In some embodiments, the modified stem cells are transiently transfected with artificial chromosomes, viruses, or plasmid DNA. In some embodiments, the virus is an oncolytic virus. In some embodiments, the virus is a vaccinia virus. In some embodiments, the virus is a replicating oncolytic vaccinia virus (VACV). In some embodiments, the modified stem cells are capable of localizing to disease sites in the subject. In some embodiments, the modified stem cells are autologous. In some embodiments, the modified stem cells are allogeneic.

[0080] IMMSTEM TMThe cells are umbilical cord-derived mesenchymal-like cells with pluripotent differentiation capacity, characterized by unique surface markers and the production of growth factors. Compared to other stem cell sources, IMMSTEM... TM IMMSTEM cells possess numerous advantages, including ease of collection, higher proliferation rates, extremely low immunogenicity, and the ability to differentiate into tissues representing all three germ layer components. Compared to other mesenchymal stem cell (MSC) subtypes, IMMSTEM... TM Cells exhibited upregulated anti-inflammatory and migration capabilities due to a "cytokine initiation" step prior to application. IMMSTEM TM The cells are derived from the human umbilical cord, which is retrieved immediately after delivery from a full-term woman. To stimulate a stress response, the cells are cultured with interferon-γ for approximately 48 hours. In some embodiments, the culture time with IFN-γ can be from 1 to 72 hours, or any value or subrange thereof.

[0081] In some implementations, stem cells are infected with poxviruses. Without being bound by theory, it is believed that poxvirus infection of stem cells can cause the cells to produce additional poxviruses, which will then be targeted to the affected areas.

[0082] In some implementations, the stem cells are not infected with poxvirus. In these implementations, the poxvirus-free stem cells have been modified to express therapeutic molecules. In these implementations, poxvirus-infected stem cells and uninfected stem cells modified to express therapeutic molecules are administered to a subject. Without being bound by theory, it is believed that the infected stem cells will target the poxvirus to the affected area, while the stem cells expressing the therapeutic molecule will target the therapeutic molecule to the affected area.

[0083] U.S. Patent No. 10,105,436, the entire contents of which are incorporated herein by reference, describes poxviruses, including smallpox vaccines, that can be used in the methods and compositions described herein.

[0084] Adipose matrix vascular grade

[0085] In some embodiments, this document discloses a method for treating a disease in a subject, comprising administering poxvirus and adipose-derived stromal vascular fraction (SVF) to the subject simultaneously, wherein the disease is not cancer. In some embodiments, the adipose-derived SVF is autologous. In some embodiments, the adipose-derived SVF is administered to the subject within approximately 24 hours after adipose tissue is obtained from the subject. In some embodiments, the adipose-derived SVF can be administered at any time after its collection and for up to approximately 48 hours after its collection, or at any point in time or within a time range thereafter. In some embodiments, the smallpox vaccine is administered via intravenous, intraperitoneal, intrathecal, intraventricular, intra-articular, intraventricular, intrapleural, intraparenchymal, or intraocular injection or intradermal injection, or any suitable delivery method thereof.

[0086] Adipose tissue can serve as an alternative to bone marrow as a source of stem cells and offers numerous benefits, including: a) extracting adipose-derived cells is a simpler and less invasive procedure compared to bone marrow extraction; b) adipose tissue contains a higher concentration of mesenchymal stem cells (MSCs) compared to bone marrow; c) the number of MSCs derived from adipose tissue does not decrease with age and therefore can serve as an autologous cell source for all patients; and d) in addition to MSCs with therapeutic potential, adipose tissue is also a source of unique cell populations, including endothelial cells, regulatory T cells, and monocytes / macrophages.

[0087] MSCs exhibit weak immunogenicity and immunomodulatory activity, characteristics that are conserved in MSCs from different tissues. It is believed that this weak immunogenicity allows for the survival and activity of allogeneic MSCs during therapeutic application.

[0088] SVF derived from whole fat aspiration reduces the need for extensive processing of internal cells, thereby minimizing the number of steps that could introduce contamination (Kurita et al., Plast. Reconstr. Surg. 2008, 121:1033-1041; discussion 1042-1033; Yoshimura et al., Aesthetic Plast. Surg. 2008, 32:48-55; discussion 56-47). The safety of fat-derived cell application is supported by autologous fat grafting, a common procedure in aesthetic medicine (Hang-Fu et al., Aesthetic Plast. Surg. 1995, 19:427-437). Each of the aforementioned references is incorporated herein by reference in its entirety.

[0089] In some embodiments, fat-derived SVFs are obtained using methods and knowledge known to those skilled in the art. In some embodiments, they are obtained via TIMEMACHINE.TM The device removes fat-derived SVF from the subject. In some embodiments, fat-derived SVF is removed from the subject using a 2.5 to 3 mM cannula. In some embodiments, one or more of the following devices may be used: a multi-functional station of PNC, CHA Biotech Cha-Station, Cytori Celution800 / CRS system, and Medi-Khan's Lipokit equipped with MaxStem.

[0090] In some embodiments, poxvirus and adipose-derived SVF are administered simultaneously. In some embodiments, poxvirus and adipose-derived SVF are administered concurrently. In some embodiments, poxvirus and adipose-derived SVF are administered concurrently via an administration medium. In some embodiments, poxvirus and adipose-derived SVF are administered concurrently via intravenous, intraperitoneal, intrathecal, intra-articular, intraocular, intraventricular, intrapleural, intraparenchymal, or intradermal injection, or any suitable delivery method thereof, through a container (e.g., a syringe).

[0091] U.S. Patent No. 10,105,436, the entire contents of which are incorporated herein by reference, describes stem cells and SVFs that can be used in the methods and compositions described herein, including methods for obtaining / preparing them.

[0092] IV. Instructions for Use

[0093] Inflammatory diseases

[0094] On one hand, this article provides a method for treating chronic inflammatory diseases in subjects with this need, the method comprising administering a poxvirus (e.g., smallpox virus, such as a smallpox vaccine) and stem cells to the subject, wherein the disease is not cancer. In an embodiment, the chronic inflammatory disease is an autoimmune disease. In an embodiment, the chronic inflammatory disease is asthma, chronic peptic ulcer, tuberculosis, arthritis, periodontitis, ulcerative colitis, Crohn's disease, sinusitis, active hepatitis, atherosclerosis, dermatitis, inflammatory bowel disease (IBS), systemic lupus erythematosus, fibromyalgia, type 1 diabetes, psoriasis, multiple sclerosis, Addison's disease, Grave's disease, Sjögren's syndrome, Hashimoto's thyroiditis, myasthenia gravis, vasculitis, pernicious anemia, and celiac disease. In one implementation plan, inflammatory diseases include transplant rejection, Dupytren's contracture, peyronies, periodontitis, endometriosis, hepatitis, glomerulonephritis, arthritis (e.g., osteoarthritis, rheumatoid arthritis, or psoriatic arthritis), inflammatory encephalopathy (including post-stroke encephalitis), and atherosclerosis. In another implementation plan, autoimmune diseases include myasthenia gravis (MG), Hashimoto's thyroiditis, vasculitis, Gray's disease, psoriasis, chronic inflammatory demyelinating polyneuropathy (CIDP), Guillain-Barré syndrome, type 1 diabetes, lupus, multiple sclerosis, rheumatoid arthritis, Addison's disease, Sjögren's syndrome, celiac disease, myositis, ankylosing spondylitis, or scleroderma.

[0095] In the implementation scheme, the inflammatory disease is an intestinal fistula, chronic radiation injury (which results in inflammatory tissue defects, such as radiation cystitis or radiation enteritis), duodenal ulcer, or a chronic inflammatory disease of the central nervous system, such as post-stroke neuroinflammation, schizophrenia, autism, addiction, chronic traumatic encephalopathy, or vaccine-induced neurotoxicity.

[0096] In one implementation, the chronic inflammatory disease is transplant rejection, Dupytren's contracture, peyronies, periodontitis, endometriosis, hepatitis, glomerulonephritis, atherosclerosis, cardiovascular disease, arthritis (e.g., osteoarthritis, rheumatoid arthritis, or psoriatic arthritis), inflammatory encephalopathy (including post-stroke, encephalitis), atherosclerosis, traumatic injury, infection, and / or shock. In one implementation, the inflammatory disease is chronic obstructive pulmonary disease (COPD), such as emphysema, chronic bronchitis, or refractory (irreversible) asthma.

[0097] In one implementation scheme, the autoimmune disease is myasthenia gravis (MG), Hashimoto's thyroiditis, vasculitis, Gray's disease, psoriasis, chronic inflammatory demyelinating polyneuropathy (CIDP), Guillain-Barré syndrome, type 1 diabetes, lupus, multiple sclerosis, rheumatoid arthritis, Addison's disease, Sjögren's syndrome, celiac disease, myositis, ankylosing spondylitis, or scleroderma.

[0098] In the implementation plan, inflammatory diseases are enterocutaneous fistulas, chronic radiation injuries (which result in inflammatory tissue defects, such as radiation cystitis or radiation enteritis), duodenal ulcers, or chronic inflammatory diseases of the central nervous system, such as post-stroke neuroinflammation, schizophrenia, autism, addiction, chronic traumatic encephalopathy, or vaccine-induced neurotoxicity.

[0099] In the implementation plan, the disease is an infectious disease, traumatic injury, and / or shock.

[0100] In the implementation scheme, the therapeutic molecule (therapeutic agent) is administered to a subject. Preferably, the therapeutic molecule (therapeutic agent) treats the disease. The therapeutic molecule (therapeutic agent) may be administered as part of a poxvirus / stem cell composition and / or separately. When the therapeutic molecule (therapeutic agent) is administered as part of a poxvirus / stem cell composition, the therapeutic molecule (therapeutic agent) may be a separate component of the composition. Alternatively (or additionally), the therapeutic molecule may be expressed by stem cells and / or encoded by a poxvirus.

[0101] In the implementation plan, the therapeutic molecules (therapeutic agents) are the therapeutic agents listed in Table 1:

[0102] Table 1: Chronic Inflammatory Diseases and Therapeutic Agents

[0103]

[0104]

[0105]

[0106] In the implementation plan, the therapeutic molecule (therapeutic agent) is selected from Orencia, Humira, Kinetet, Cimzia, Enbrel, Simponi, Remicade, Taltz, Tysabri, Rituxan, Cosentyx, Actemra, Stelara, Entyvio, Simulect, Zinbryta, and Orthoclone OKT3.

[0107] In the implementation scheme, the therapeutic molecules include TGFβ, HGF, LIF, VEGF, EGF, BDNF and / or NGF, or fragments thereof.

[0108] In the implementation scheme, the therapeutic molecule is an antibiotic. Antibiotics are well known in the art. The antibiotic can be any antibiotic. A skilled clinician can determine which antibiotic should be used based on the type of infection and other criteria. Non-limiting examples of antibiotics are actinomycin, bacitracin, colistin, polymyxin B, brevicin, polymyxins, bacitracin derivatives, glycopeptides, etc.

[0109] In one aspect, this document provides a method for converting chronic inflammation into acute inflammation in a subject with this need. The method includes administering a poxvirus to the subject, wherein the condition is not cancer. In an embodiment, the poxvirus is administered in a therapeutically effective amount, for example, an amount sufficient to convert chronic inflammation into acute inflammation. In an embodiment, the method also includes treating the acute inflammation. In an embodiment, treating the acute inflammation includes administering a known treatment for acute inflammation to the subject.

[0110] In one embodiment, the disease is interstitial cystitis. Without being bound by theory, anti-proliferative factor (APF) is believed to inhibit bladder cell proliferation by regulating the production of cell adhesion proteins and growth factors. In one embodiment, the therapeutic agent expressed by a virus and / or stem cells is a polypeptide that blocks APF expression and / or activity. In one embodiment, the therapeutic agent expressed by a virus and / or stem cells is a polypeptide that blocks NF-κB activity. In one embodiment, the therapeutic agent expressed by a virus and / or stem cells is a polypeptide that blocks aberrant NF-κB activity. In one embodiment, the therapeutic agent is HB-EGF or a fragment or variant thereof. See, for example, Kim et al., BJU Int. Feb. 2009; 103(4):541–546, the entire contents of which are incorporated herein by reference.

[0111] In this implementation, the disease is atherosclerosis. Without being bound by theory, it is predicted that inducing eNOS on the endothelial surface will alleviate atherosclerosis and may even reverse plaque formation. In one implementation, the therapeutic agent expressed by a virus and / or stem cells is a peptide that induces eNOS.

[0112] In one implementation, an infectious disease causes or is capable of causing a cytokine storm in the subject. In one implementation, poxvirus and optionally stem cells are administered to treat the cytokine storm.

[0113] Infectious diseases

[0114] In one aspect, this article provides a method for treating an infectious disease in a subject with this need. The method includes administering a poxvirus to the subject. In this implementation, the disease is not cancer.

[0115] Infectious diseases can be caused by any organism known to be capable of infecting a subject. In embodiments, infectious diseases are caused by bacteria, viruses, parasites, protozoa, worms, or fungi. In embodiments, infectious diseases are caused by bacteria. In embodiments, infectious diseases are caused by viruses. In embodiments, infectious diseases are caused by parasites. In embodiments, infectious diseases are caused by fungi. In embodiments, infectious diseases can be caused by one or more media listed in U.S. Patent No. 8,715,677 and / or U.S. Patent Publication No. 2019 / 0381160, the entire contents of which are incorporated herein by reference.

[0116] In this implementation, the virus is a virus that infects humans. In this implementation, the virus causes respiratory infections, gastrointestinal infections, foodborne illnesses, skin infections, or sexually transmitted infections. In this implementation, the virus is a rhinovirus, coronavirus, influenza, or respiratory syncytial virus. In this implementation, the virus is a coronavirus. In this implementation, the coronavirus is Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2).

[0117] In this implementation, stem cells are administered to a subject. The stem cells can be any type of stem cell, such as those described herein. In this implementation, the stem cells are mesenchymal stem cells. In this implementation, the stem cells are adipose-derived stem cells.

[0118] In the implementation scheme, the therapeutic molecule (therapeutic agent) is administered to a subject. Preferably, the therapeutic molecule (therapeutic agent) treats a disease or symptoms of a disease. The therapeutic molecule (therapeutic agent) may be administered as part of a poxvirus / stem cell composition and / or separately. When the therapeutic molecule (therapeutic agent) is administered as part of a poxvirus / stem cell composition, the therapeutic molecule (therapeutic agent) may be a separate component of the composition. Alternatively (or additionally), the therapeutic molecule may be expressed by stem cells and / or encoded by a poxvirus.

[0119] In the implementation scheme, the therapeutic molecule is an antibiotic. Antibiotics are well known in the art. The antibiotic can be any antibiotic. A skilled clinician can determine which antibiotic should be used based on the type of infection and other criteria. Non-limiting examples of antibiotics are actinomycin, bacitracin, colistin, polymyxin B, brevicin, polymyxins, bacitracin derivatives, glycopeptides, etc.

[0120] In one embodiment, the therapeutic molecule is an antiviral molecule. In another embodiment, the therapeutic molecule is an antifungal molecule. In yet another embodiment, the therapeutic molecule is an antiparasitic molecule. In one embodiment, the therapeutic molecule is a vaccine. In another embodiment, the therapeutic molecule comprises molecules with anti-inflammatory and / or nutritional activity. In another embodiment, the therapeutic molecule is TGFβ, HGF, LIF, VEGF, EGF, BDNF, and / or NGF, or fragments thereof. In yet another embodiment, the therapeutic molecule is a molecule that treats or prevents cytokine storms, such as leukemia suppressor factor (LIF).

[0121] In one implementation, an infectious disease causes or is capable of causing a cytokine storm in the subject. In another implementation, poxvirus and, optionally, stem cells are administered to treat the cytokine storm.

[0122] Cancer patients

[0123] In one aspect, this article provides a method for treating or preventing inflammatory or infectious diseases in subjects with cancer. Cancer patients often have compromised immune function, for example, due to cancer treatments (chemotherapy, radiation therapy, immunotherapy, etc.). Patients with compromised immune function are more susceptible to pathogens that cause infectious diseases.

[0124] In one implementation, the method includes administering poxvirus to a subject. In another implementation, inflammatory diseases are treated. In yet another implementation, inflammatory diseases are prevented. In one implementation, infectious diseases are treated. In yet another implementation, infectious diseases are prevented. In one implementation, stem cells are administered. Any method or composition described herein can be used to treat a subject.

[0125] Without being bound by theory, it is believed that administering poxvirus as described herein, optionally in conjunction with stem cells, will result in infection of tumor cells in a patient by the poxvirus. In one embodiment, the poxvirus encodes a therapeutic molecule. In another embodiment, stem cells express a therapeutic molecule. In yet another embodiment, the therapeutic molecule treats or prevents inflammatory diseases. In yet another embodiment, the therapeutic molecule treats or prevents infectious diseases. In yet another embodiment, the therapeutic molecule is a vaccine for a disease. In yet another embodiment, the therapeutic molecule is an antibiotic. In yet another embodiment, the therapeutic molecule is an antiviral molecule. In yet another embodiment, the therapeutic molecule is an antifungal molecule. In yet another embodiment, the therapeutic molecule is an antiparasitic molecule.

[0126] In the implementation plan, infecting tumor cells with poxviruses causes the tumor cells to express therapeutic molecules. Without being bound by theory, it is believed that the expression of therapeutic molecules by tumor cells leads to an increase in the amount of therapeutic molecules delivered to the subject, allowing for the treatment or prevention of disease.

[0127] application

[0128] In one implementation scheme, poxvirus and / or stem cells are administered to a subject via intravenous injection, intraperitoneal injection, intrathecal injection, intraventricular injection, intra-articular injection, intracerebral injection, intrapleural injection, intraparenchymal injection, or intraocular injection. In another implementation scheme, poxvirus and optional stem cells are administered directly to the disease-affected area. In yet another implementation scheme, poxvirus and optional stem cells are administered via direct injection. In yet another implementation scheme, poxvirus and optional stem cells are administered via MRI-guided delivery.

[0129] In the implementation plan, the stem cells are autologous to the subject. That is, the stem cells are derived from the patient to be treated. For example, adipose-derived stromal vascular fractions can be taken from the patient, from which stem cells are harvested or otherwise derived.

[0130] In the implementation plan, the stem cells are allogeneic to the subject. In the implementation plan, the allogeneic stem cells are derived from a subject other than the patient. In the implementation plan, the allogeneic stem cells are derived from a cell line.

[0131] In the implementation plan, the subjects are humans.

[0132] In some embodiments, poxvirus and stem cells are administered simultaneously. In some embodiments, poxvirus and stem cells are administered simultaneously via an administration medium. In some embodiments, poxvirus and stem cells are administered simultaneously via intravenous, intraperitoneal, intrathecal, intracardiac, intra-articular, intraocular, intraventricular, intrapleural, intraparenchymal, or intradermal injection, or any suitable delivery method thereof, through a container (e.g., a syringe).

[0133] The effective dosage of each treatment method disclosed herein can vary depending on various factors, including but not limited to the specific treatment method employed, the compound or pharmaceutical composition, the administration method, the condition being treated, and / or the severity of the condition. Therefore, the dosage regimen of the combinations of the present invention is selected based on multiple factors, including the route of administration and the patient's renal and hepatic function. A physician, clinician, or veterinarian with ordinary skills can easily determine and prescribe an effective amount of a single active ingredient required to prevent, counteract, or halt the progression of a condition. To achieve optimal concentration of the active ingredient and ensure its efficacy without toxicity, the regimen needs to be developed based on the bioavailability kinetics of the active ingredient at the target site.

[0134] The amount of poxvirus administered to an average-sized adult can be, for example, 1 x 10^6. 2 Up to 1x10 10 Each plaque forming unit, 1x10 3 Up to 1x10 8 Each plaque forming unit, 1x10 4 Up to 1x10 6 Each plaque formation unit, or any value or subrange in between. As a concrete example, approximately 2.5 x 10-1 could be used. 5 Each empty spot forming unit.

[0135] It should be understood that the implementation schemes described herein are not limited to vaccination or the administration of vaccines themselves, but also involve generating an immune response or reaction to disease-related antigens. While the terms "vaccine," "vaccination," or other similar terms are used for convenience, it should be understood that these implementation schemes also involve immune compositions, immunogenic compositions, the generation of an immune response, immunization, etc., where absolute preventive immunity is not required or generated. For example, implementation schemes mentioning vaccination may also involve generating or assisting in the generation of an immunogenic or immune response against an antigen, regardless of whether that response results in absolute eradication or immunity against the disease to be treated.

[0136] U.S. Patent No. 10,105,436, the entire contents of which are incorporated herein by reference, describes methods for applying, preparing, storing, and using compositions comprising poxvirus and stem cells, which can be used in the methods and compositions described herein.

[0137] V. Composition

[0138] On the one hand, this article provides compositions comprising a poxvirus (e.g., smallpox virus, such as a smallpox vaccine) and optional stem cells, wherein the poxvirus comprises a recombinant polynucleotide, wherein the recombinant polynucleotide encodes a therapeutic molecule.

[0139] On the one hand, this article provides compositions comprising poxvirus (e.g., smallpox virus, such as smallpox vaccine) and stem cells, wherein the stem cells comprise recombinant polynucleotides, wherein the recombinant polynucleotides encode therapeutic molecules.

[0140] In one implementation scheme, the therapeutic molecule treats a chronic inflammatory disease. In another implementation scheme, the therapeutic molecule treats an autoimmune disease. In yet another implementation scheme, the therapeutic molecule treats an infectious disease, traumatic injury, or shock. In this implementation scheme, the therapeutic molecule is a cytokine, a therapeutic antibody, a therapeutic fusion protein, an antibiotic, RNA, a nucleotide, a peptide, or a polypeptide. In this implementation scheme, the cytokine is selected from interleukin (IL)-1 receptor antagonists, IL-4, IL-6, IL-10, IL-11, IL-13, IFN-α, and transforming growth factor-β.

[0141] On one hand, this document provides compositions comprising poxviruses (e.g., smallpox virus, such as smallpox vaccine), optional stem cells, and therapeutic agents. In one embodiment, the poxvirus comprises a recombinant polynucleotide, wherein the recombinant polynucleotide encodes a therapeutic molecule. In one embodiment, the stem cells comprise a recombinant polynucleotide, wherein the recombinant polynucleotide encodes a therapeutic molecule.

[0142] In the implementation plan, the therapeutic agents are those listed in Table 1. Specifically, the therapeutic agents are abatacept (Orencia), adalimumab (Humira), anaspirin (Kineret), cetuzumab (Cimzia), etanercept (Enbrel), golimumab (Simponi), infliximab (Remicade), taltz, nateximab (Taltz), rituximab (Rituxan), secukinumab (Cosentyx), tocilizumab (Actemra), stelara, entyvio, baliximab (Simulect), daxizumab (Zinbryta), or morocumab (Orthoclone OKT3).

[0143] In the implementation plan, RNA is antisense RNA, siRNA, RNA vaccine, miRNA, or RNA interference (RNAi).

[0144] Antibiotics are well-known in this field. Skilled clinicians can determine which antibiotics to use based on the type of infection and other standard assays.

[0145] Stem cells can be any type of stem cell, especially those described in this article.

[0146] The poxvirus can be any poxvirus, especially the poxvirus described herein. In this implementation, the poxvirus is an attenuated virus.

[0147] Methods for preparing pharmaceutical compositions comprising the relevant therapeutic means disclosed herein are known in the art and are apparent from the prior art and from known standard references such as Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, Pa., 18th edition (1990), the entire contents of which are incorporated herein by reference.

[0148] In some embodiments, the compositions disclosed herein comprise a pharmaceutically acceptable carrier. As used herein, the term "pharmaceuticalally acceptable carrier" refers to a solvent, diluent, preservative, dispersant or suspending agent, isotonic agent, thickener or emulsifier, solid binder, and lubricant suitable for a particular dosage form. Those skilled in the art are familiar with the various different carriers that can be used to formulate pharmaceutical compositions and with their preparation techniques (see Remington's Pharmaceutical Sciences, edited by Gennaro, Mack Publishing, Easton, Pa., 1995; the entire contents of which are incorporated herein by reference). Pharmaceutically acceptable carriers may include, but are not limited to, Ringer's solution, isotonic saline, starch, potato starch, sugars, glucose, astragalus gum powder, malt, gelatin, talc, cellulose and its derivatives, ethyl cellulose, sodium carboxymethyl cellulose, cellulose acetate excipients, cocoa butter, suppository waxes, agar, alginic acid, oils, cottonseed oil, peanut oil, safflower oil, sesame oil, olive oil, soybean oil, corn oil, glycols, propylene glycol, esters, ethyl laurate, ethyl oleate, buffers, aluminum hydroxide, magnesium hydroxide, phosphate buffer, pyrogen-free water, ethanol, other non-toxic and compatible lubricants, sodium dodecyl sulfate, magnesium stearate, colorants, releasing agents, coating agents, sweeteners, flavoring agents, and aroma agents. Pharmaceutically acceptable carriers may also include preservatives and antioxidants. One or more of the above substances may be specifically excluded from the compositions and methods of some embodiments.

[0149] The poxvirus-containing compositions disclosed herein may include adjuvants. Optionally, the composition may include one or more compounds having adjuvant activity. Adjuvants are nonspecific stimulants of the immune system that enhance the host's immune response to a vaccine. Examples of adjuvants known in the art are Freund's complete and incomplete adjuvants, vitamin E, nonionic block polymers, muramyl dipeptides, ISCOM (immunostimulatory complex), saponins, mineral oils, vegetable oils, and carbopol. Adjuvants particularly suitable for mucosal application are, for example, Escherichia coli heat-sensitive toxin (LT) or cholera toxin (CT). Other suitable adjuvants are, for example, aluminum hydroxide, aluminum phosphate or aluminum oxide, oil emulsions (e.g., Bayol). Or Marcol (Oil emulsions, saponins, or vitamin E solubilizers). One or more of the above substances may be specifically excluded from the compositions and methods of some embodiments.

[0150] U.S. Patent No. 10,105,436, the entire contents of which are incorporated herein by reference, describes methods for applying, preparing, storing, and using compositions comprising poxvirus and stem cells, which can be used in the methods and compositions described herein.

[0151] It should be understood that the examples and embodiments described herein are for illustrative purposes only, and those skilled in the art will make various modifications or changes based on these examples and embodiments, which will be included within the spirit and scope of this application and the appended claims. All publications, patents, and patent applications cited herein are incorporated herein by reference in their entirety for whatever purpose. Example

[0152] Example 1: Preparation of ACAM2000 vaccine

[0153] Before reconfiguration, the vaccine vials should be removed from the cold storage and placed at room temperature. Remove the flip-top seals from the vaccine vials and diluent vials, wipe each rubber stopper with an isopropyl alcohol swab, and allow it to dry completely.

[0154] Using aseptic technique and a 1mL sterile syringe fitted with a 25 x 5 / 8" needle, transfer 0.3mL of diluent to the vaccine vial. Gently vortex the vial to mix, preferably avoiding contact with the rubber stopper. The reconstituted vaccine should be a clear to slightly turbid, colorless to pale yellow liquid, free of unwanted substances. Before administration, visually inspect the reconstituted vaccine for particulate matter and discoloration. If particulate matter or discoloration is observed, discontinue use and dispose of the vial safely.

[0155] After reconstitution, each vial contains approximately 2.5–12.5 x 10⁷ plaque-forming units (PFU) of live vaccinia virus. After reconstitution, the ACAM2000 vaccine can be used within 6–8 hours if kept at room temperature (20–25°C, 68–77°F). Unused reconstituted ACAM2000 vaccine can be stored in a refrigerator (2–8°C, 36–46°F) for up to 30 days, after which it should be discarded as biohazardous material. Personnel preparing and administering the vaccine should wear surgical or protective gloves to avoid contact with skin, eyes, or mucous membranes. Vaccine vials, their stoppers, diluents, syringes, venting needles used for reconstitution, needles used for administration, and any materials that have come into contact with the vaccine should be discarded in leak-proof, puncture-proof biohazard containers. These containers should then be disposed of appropriately.

[0156] When applying the vaccine, gently vortex the vial to mix, preferably avoiding contact with the rubber stopper. Using aseptic technique and a 1mL sterile syringe fitted with a 25 x 5 / 8 inch needle, transfer the entire contents of the vial to a labeled 20cc syringe containing the SVF fraction. Gently vortex the SVF-vaccine mixture to thoroughly mix and incubate at 37°C for 2 to 4 hours.

[0157] Example 2: Acquisition and preparation of adipose matrix vascular fractions

[0158] The patient received local anesthesia consisting of 0.5% lidocaine and epinephrine (1:400,000), titrated to pH 7.4 with 8.4% HCO3 (generally 5 cc of HCO3 is added to a total volume of 60 cc), and aseptic preparation was performed. Then, using, for example, TIME-MACHINE... TM The device, fat processor (syringe), and 2.5-3mm cannula are used to perform liposuction on the patient. Bacitracin ointment and adhesive bandages are applied to the wound, which is then secured with a pressure bandage.

[0159] Prepare SVF (ADSC) in a closed system according to the following scheme:

[0160] a. will The collected fat was placed into a 60cc disposable sterile fat processing syringe.

[0161] b. Centrifuge the fat at 2800 rpm for 3 minutes.

[0162] c. Removal of free fatty acids and debris (locally / in the blood) via the TP-109 closed system.

[0163] d. Transfer 25cc of concentrated fat to an SVF treatment syringe.

[0164] e. Add 25cc of preheated (38℃) T- Time Machine Accelerator (GMP-grade collagenase; Roche), containing 12.5 Wunsch units.

[0165] f. Incubate at 38°C for 30-45 minutes.

[0166] g. Centrifuge at 200 x g for 4 minutes

[0167] h. Remove the supernatant except for the bottom 3 to 10 cc.

[0168] i. Add 50cc of D5LR as washing buffer to remove collagenase residue, and centrifuge at 200x g for 4 minutes.

[0169] j. Repeat 2 more times, for a total of 3 washes.

[0170] k. Remove all supernatant, leaving 3 to 10 cc of precipitate for collection—this is the stromal vascular fraction.

[0171] 1. Filter the SVF into a 20cc syringe through a 100-micron filter.

[0172] m. Collect SVF samples and identify cell number and viability, and confirm that there are no clumps or debris.

[0173] n. Aliquots of each cell suspension are reserved for endotoxin testing and sterile staining. SVF injection will only proceed after confirming that the endotoxin test result is less than or equal to 5 EU / kg / hr and the Gram staining result is negative.

[0174] o. Resuspend the cells in 20 ml of physiological saline. Aspirate the cell suspension into a syringe through an 18-gauge needle for injection. Up to 100 million live cells will be used for injection.

[0175] p. Then, the syringe is placed in a sealed sample bag labeled with the patient's name and medical record number for transport to the operating room for injection.

[0176] Example 3: Application of Fat SVF and Smallpox Vaccine

[0177] Application method:

[0178] Intravenous: Non-amplified, autologous matrix vascular fraction (SVF) containing up to 100 million cells, extracted from up to 500 ml of fat aspirate and purified by collagenase digestion and a series of washing steps, is incubated with vaccinia virus and delivered in a 20 ml volume via intravenous injection.

Claims

1. The use of a combination of attenuated poxvirus and stem cells in the preparation of a medicament for treating a chronic inflammatory disease in subjects of need, wherein the chronic inflammatory disease is not cancer. The stem cells and / or poxviruses contained in the poxviruses contained recombinant polynucleotides, wherein the recombinant polynucleotides encoded therapeutic molecules for treating the chronic inflammatory disease.

2. Use of the combination of attenuated poxvirus and stem cells in the preparation of a medicament for treating an infectious disease in subjects in need, wherein the infectious disease is not cancer. The stem cells and / or poxviruses contained in the poxviruses contained recombinant polynucleotides, wherein the recombinant polynucleotides encoded therapeutic molecules for treating the infectious diseases.

3. The use according to claim 1, wherein the chronic inflammatory disease is an autoimmune disease.

4. The use as described in claim 1 or 3, wherein the chronic inflammatory disease is selected from asthma, chronic peptic ulcer, tuberculosis, arthritis, periodontitis, ulcerative colitis, Crohn's disease, sinusitis, active hepatitis, atherosclerosis, dermatitis, inflammatory bowel disease (IBS), systemic lupus erythematosus, fibromyalgia, type 1 diabetes, psoriasis, multiple sclerosis, Addison's disease, Grave's disease, Sjögren's syndrome, Hashimoto's thyroiditis, myasthenia gravis, vasculitis, pernicious anemia, or celiac disease.

5. The use according to claim 3, wherein the autoimmune disease is myasthenia gravis (MG), Hashimoto's thyroiditis, vasculitis, Gray's disease, psoriasis, chronic inflammatory demyelinating polyneuropathy (CIDP), Guillain-Barré syndrome, type 1 diabetes, lupus, multiple sclerosis, rheumatoid arthritis, Addison's disease, Sjögren's syndrome, celiac disease, myositis, ankylosing spondylitis, or scleroderma.

6. The use as described in claim 1 or 3, wherein the chronic inflammatory disease is transplant rejection, Dupytren's contracture, peyronies, periodontitis, endometriosis, hepatitis, glomerulonephritis, atherosclerosis, cardiovascular disease, arthritis (e.g., osteoarthritis, rheumatoid arthritis, or psoriatic arthritis), inflammatory encephalopathy (including post-stroke, encephalitis), atherosclerosis, traumatic injury, infection, chronic obstructive pulmonary disease (COPD), and / or shock.

7. The use according to claim 1 or 3, wherein the inflammatory disease is an intestinal fistula, chronic radiation injury (which results in inflammatory tissue defects, such as radiation cystitis or radiation enteritis), duodenal ulcer, or a chronic inflammatory disease of the central nervous system, such as post-stroke neuroinflammation, schizophrenia, autism, addiction, chronic traumatic encephalopathy, or vaccine-induced neurotoxicity.

8. The use according to claim 6, wherein the COPD is emphysema, chronic bronchitis or refractory (irreversible) asthma.

9. The use according to claim 2, wherein the infectious disease is caused by bacteria, viruses or fungi.

10. The use according to claim 9, wherein the infectious disease is caused by a virus.

11. The use as claimed in claim 10, wherein the virus is rhinovirus, coronavirus, influenza, or respiratory syncytial virus.

12. The use according to claim 11, wherein the coronavirus is severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2).

13. The use according to any one of claims 1-12, wherein the poxvirus is a vaccinia virus.

14. The use as claimed in claim 13, wherein the vaccinia virus is selected from Dryvax, ACAM1000, ACAM2000, Lister, EM63, LIVP, Tian Tan, Copenhagen, Western Reserve, modified vaccinia virus Ankara (MVA), New York City Board of Health, Dairen, Ikeda, LC16M8, Western Reserve Copenhagen, Tashkent, Wyeth, IHD-J and IHD-W, Brighton, Dairen I and Connaught strains.

15. The use as claimed in claim 14, wherein the vaccinia virus is ACAM1000 or ACAM2000.

16. The use as claimed in claim 14, wherein the vaccinia virus is a New York City Department of Health strain.

17. The use according to any one of claims 1-16, wherein the therapeutic molecule is a cytokine, a therapeutic antibody, a therapeutic fusion protein, an antibiotic, RNA, a receptor that promotes the uptake of a therapeutic agent, an antigen recognized by the therapeutic agent, or an enzyme used by the cell to produce the therapeutic agent.

18. The use according to any one of claims 1-16, wherein the therapeutic molecule is selected from anti-TNF antibodies, T-cell receptor-directed antibodies, IL-2 receptor-directed antibodies, and interferons.

19. The use of any one of claims 1-16, wherein the therapeutic molecule is selected from abatacept (Orencia), adalimumab (Humira), anaphylactin (Kineret), cetuzumab (Cimzia), etanercept (Enbrel), golimumab (Simponi), infliximab (Remicade), taltz (Taltz), nateximab (Tysabri), rituximab (Rituxan), cosentyx (Cosentyx), tocilizumab (Actemra), stelara (Stelara), entyvio (Entyvio), baliximab (Simulect), daxizumab (Zinbryta), and morocumab (Orthoclone OKT3).

20. The use according to any one of the preceding claims, wherein the treatment further comprises administering a therapeutic agent to the subject.

21. The use according to claim 20, wherein the therapeutic agent is a medicine for treating chronic inflammatory diseases.

22. The use according to claim 20 or 21, wherein the therapeutic agent is selected from 5-aminosalicylate / salt, corticosteroids, azathioprine, mercaptopurine, cyclosporine, bronchodilators, roflumilast, statins, fibrates, beta-blockers, ACE inhibitors, diuretics, aspirin, calcium channel blockers, collagenase, verapamil, topical antibacterial agents, penicillin, antibiotics, hormones, hepatitis A vaccine, hepatitis B vaccine, immunosuppressants, cyclophosphamide, NSAIDs, analgesics, anesthetics, steroids, proton pump inhibitors, antiviral drugs, anticonvulsants, blood thinners, antihypertensive drugs, and adrenaline.

23. The use as claimed in claim 20 or 21, wherein the therapeutic agent is selected from abatacept (Orencia), adalimumab (Humira), anaphylactin (Kineret), cetuzumab (Cimzia), etanercept (Enbrel), golimumab (Simponi), infliximab (Remicade), taltz (Taltz), nateximab (Tysabri), rituximab (Rituxan), cosentyx (Cosentyx), tocilizumab (Actemra), stelara (Stelara), entyvio (Entyvio), baliximab (Simulect), daxizumab (Zinbryta), and morocumab (Orthoclone OKT3).

24. The use according to claim 20, wherein the therapeutic agent is a medicine for treating viral infection or its symptoms.

25. The use according to claim 1, wherein the stem cells are selected from adult stem cells, embryonic stem cells, fetal stem cells, mesenchymal stem cells, neural stem cells, totipotent stem cells, pluripotent stem cells, multipotent stem cells, oligopotent stem cells, unipotent stem cells, adipose stromal cells, endothelial stem cells, induced pluripotent stem cells, bone marrow stem cells, umbilical cord blood stem cells, adult peripheral blood stem cells, myoblast stem cells, juvenile small stem cells, skin fibroblast stem cells, and combinations thereof.

26. The use according to claim 25, wherein the stem cell is an adipose stem cell.

27. The use according to any one of claims 1-26, wherein the stem cell is a stem cell line or is derived from a stem cell line.

28. The use according to any one of claims 1-27, wherein the stem cells are modified stem cells.

29. The use of claim 28, wherein the modified stem cells express heterologous proteins.

30. The use of claim 29, wherein the heterologous protein is a therapeutic molecule.

31. The use according to claim 30, wherein the therapeutic molecule treats the disease.

32. The use as described in claim 30 or 31, wherein the therapeutic molecule is a cytokine, a therapeutic antibody, a therapeutic fusion protein, an antibiotic, RNA, a receptor that promotes the uptake of a therapeutic agent, an antigen recognized by the therapeutic agent, or an enzyme used by the cell to produce the therapeutic agent.

33. The use according to any one of claims 30-32, wherein the therapeutic molecule is selected from anti-TNF antibodies, T-cell receptor-directed antibodies, IL-2 receptor-directed antibodies, and interferons.

34. The use according to any one of claims 30-33, wherein the therapeutic molecule is selected from abatacept (Orencia), adalimumab (Humira), anaphylactin (Kineret), cetuzumab (Cimzia), etanercept (Enbrel), golimumab (Simponi), infliximab (Remicade), taltz (Taltz), nateximab (Tysabri), rituximab (Rituxan), cosentyx (Cosentyx), tocilizumab (Actemra), stelara (Stelara), entyvio (Entyvio), baliximab (Simulect), daxizumab (Zinbryta), and morocumab (Orthoclone OKT3).

35. The use according to any one of claims 1-34, wherein the poxvirus and / or the stem cells are administered to the subject by intravenous, intraperitoneal, intrathecal, intraventricular, intrapleural, intraparenchymal, intracardiac, intraarticular, or intraocular injection.

36. The use according to any one of claims 1-34, wherein the poxvirus and / or the stem cells are applied directly to the area affected by the disease.

37. The use as claimed in claim 36, wherein the poxvirus and / or stem cells are administered via MRI-guided delivery.

38. The use of any of the preceding claims, wherein the stem cells are autologous to the subject.

39. The use according to any one of claims 1-37, wherein the stem cells are allogeneic to the subject.

40. The use of any of the preceding claims, wherein the subject is a human.

41. The use according to any one of claims 1-39, wherein the subject is a domesticated animal.

42. The use according to any one of claims 1-39, wherein the subject is a companion animal.

43. The use according to claim 42, wherein the subject is a canine.

44. A composition comprising stem cells and an attenuated poxvirus, wherein the poxvirus comprises a recombinant polynucleotide, wherein the recombinant polynucleotide encodes a therapeutic molecule capable of treating inflammatory or infectious diseases.

45. The composition of claim 44, wherein the therapeutic molecule treats an inflammatory disease.

46. ​​The composition of claim 44 or 45, wherein the therapeutic molecule is a cytokine, a therapeutic antibody, a therapeutic fusion protein, RNA, an antibiotic, a receptor that promotes the uptake of the therapeutic agent, an antigen recognized by the therapeutic agent, or an enzyme used by the cell to produce the therapeutic agent.

47. The composition of claim 44 or 45, wherein the therapeutic molecule is selected from anti-TNF antibodies, T-cell receptor-directed antibodies, IL-2 receptor-directed antibodies, and interferons.

48. The composition of claim 44 or 45, wherein the therapeutic molecule is selected from abatacept (Orencia), adalimumab (Humira), anaphylactin (Kineret), cetuzumab (Cimzia), etanercept (Enbrel), golimumab (Simponi), infliximab (Remicade), taltz (Taltz), nateximab (Tysabri), rituximab (Rituxan), cosentyx (Cosentyx), tocilizumab (Actemra), stelara (Stelara), entyvio (Entyvio), baliximab (Simulect), daxizumab (Zinbryta), and morocumab (Orthoclone OKT3).

49. The composition of any one of claims 44-48, wherein the poxvirus is a vaccinia virus.

50. The composition of claim 49, wherein the vaccinia virus is selected from Dryvax, ACAM1000, ACAM2000, Lister, EM63, LIVP, Tian Tan, Copenhagen, Western Reserve, modified vaccinia virus Ankara (MVA), New York City Board of Health, Dairen, Ikeda, LC16M8, Western Reserve Copenhagen, Tashkent, Wyeth, IHD-J and IHD-W, Brighton, Dairen I and Connaught strains.

51. The composition of claim 50, wherein the vaccinia virus is ACAM1000 or ACAM2000.

52. The composition of claim 50, wherein the vaccinia virus is a New York City Department of Health strain.

53. The composition of any one of claims 44-52, wherein the stem cells comprise a recombinant polynucleotide, wherein the recombinant polynucleotide encodes a second therapeutic molecule.

54. The composition of claim 53, wherein the second therapeutic molecule is selected from abatacept (Orencia), adalimumab (Humira), anaphylactin (Kineret), cetuzumab (Cimzia), etanercept (Enbrel), golimumab (Simponi), infliximab (Remicade), taltz (Taltz), nateximab (Tysabri), rituximab (Rituxan), cosentyx (Cosentyx), tocilizumab (Actemra), stelara (Stelara), entyvio (Entyvio), baliximab (Simulect), daxizumab (Zinbryta), and morocumab (Orthoclone OKT3).

55. The composition of any one of claims 44-54, wherein the stem cells are selected from adult stem cells, embryonic stem cells, fetal stem cells, mesenchymal stem cells, neural stem cells, totipotent stem cells, pluripotent stem cells, multipotent stem cells, oligopotent stem cells, unipotent stem cells, adipose stromal cells, endothelial stem cells, induced pluripotent stem cells, bone marrow stem cells, umbilical cord blood stem cells, adult peripheral blood stem cells, myoblast stem cells, juvenile small stem cells, skin fibroblast stem cells, and combinations thereof.

56. The composition of any one of claims 44-55, wherein the stem cells are modified stem cells.

57. The composition of claim 56, wherein the modified stem cells express heterologous proteins.

58. The composition of any one of claims 44-57, wherein the stem cells are derived from a subject to be treated with the composition.

59. The composition of any one of claims 44-58, wherein the stem cells are derived from humans.

60. The composition of any one of claims 44-58, wherein the stem cells are derived from domesticated animals.

61. The composition of any one of claims 44-58, wherein the stem cells are derived from companion animals.

62. The composition of claim 61, wherein the stem cells are derived from a canine animal subject.

63. The use according to any one of claims 1-43, wherein the stem cells maintain their pluripotency or their ability to self-renew when combined with the medicament.

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