CN116583274A - Methods and pharmaceutical compositions for the treatment and prevention of microbial infectious diseases and related inflammatory disorders - Google Patents

Abstract

Pharmaceutical compositions and methods useful in the treatment and prevention of various infections, diseases and conditions are provided. Preferably, the composition and method can be used for preventing and treating aging and aging-related diseases and uses (or methods) thereof. Preferably, the compositions and methods are useful for the treatment and prevention of microbial infectious diseases and related inflammatory disorders and uses (or methods) thereof. The pharmaceutical composition according to the present invention comprises a therapeutic agent X or a pharmaceutically acceptable salt thereof; a therapeutic agent Y or a pharmaceutically acceptable salt thereof; and at least one pharmaceutically acceptable excipient, wherein the therapeutic Sexual agent X is a non-steroidal anti-inflammatory drug and the therapeutic agent Y is a fatty acid oxidation inhibitor.

Description

Methods and methods for treating and preventing microbial infectious diseases and related inflammatory disorders pharmaceutical composition

Cross References to Related Applications

This application claims the title "METHODS AND COMPOSITIONS FOR THETREATMENT AND PROPHYLAXIS OF MICROBIAL INFECTIOUS DISEASES AND ASSOCIATEDINFLAMMATORY DISORDERS," filed December 14, 2020. priority and benefit as of the filing date of U.S. Provisional Application No. 63/124,905, which is hereby incorporated by reference in its entirety. This application also claims U.S. Provisional Application Number entitled "METHODS AND COMPOSITIONS FORTHE TREATMENT AND PROPHYLAXIS OF AGING AND ASSOCIATED DISEASES," filed January 20, 2021 63/139,337, which is hereby incorporated by reference in its entirety.

technical field

This patent specification relates to the field of treatment and prevention of diseases and infections. More particularly, this patent specification relates to the use of small molecules to prevent or treat aging and related diseases and the use of small molecules to prevent or treat microbial infections.

Background technique

There are many diseases and infections that affect humans and other mammals. This includes infectious diseases and related inflammatory disorders as well as aging and related diseases.

Microbial infectious diseases are the combined process of invasion of the organism by infectious agents, replication of these agents, and host tissue responses to these agents, including inflammation, adaptive immune responses, and sometimes cytokine storms. Examples of microorganisms include viruses, bacteria, fungi, protozoa and parasites. Microorganisms replicate within host cells and can produce toxins that cause disease. Microbial infections tend to show varying degrees of severity in different infected individuals. In mild cases, infection may manifest as symptoms such as fever, chills, fatigue, cough, and diarrhea. In severe cases, it may manifest as dyspnea, rapid heart rate, tremor, chest pain, acute lung injury (ALI), and acute respiratory distress syndrome (ARDS) , cytokine release syndrome (CRS), thrombosis, tissue inflammation, tissue damage, acute organ failure, sepsis, and even death. As already discussed, the severity of microbial infection depends largely on the host response. Therefore, it will be important to target microbial infections by modulating the host's endogenous systems by modulating the host's internal environment.

As mentioned, microbial infections can be caused by viruses. Viral infections can be caused by DNA or RNA viruses, such as members of the Myoviridae, Brachyphageidae, Longobacteriidae, Heterpesviridae, Herpesviridae (including Human Herpesviruses, and Varicella-Zoster Viruses) , Malocoherpesviridae (Malocoherpesviridae), Limophageidae, Archaphageidae, Adenoviridae, Ampullaviridae (Ampullaviridae), Vesciviridae, African Swine Fever Viridae (including African Swine Fever Virus), Baculoviridae Rhabdoviridae, Cicaudaviridae, Rhabdoviridae, Covering Phages, Spindle Viridae, Spheroviridae, Trichoviridae, Hytrosaviridae, Iridoviridae, Maseilleviridae, Mimiviridae, Nudiviridae, Nimaviridae, Pandoraviridae, Papillomaviridae, Phytoviridae, Budophilic Phagedae, PolyDNA Viruses (Polydnaviruses), Polyomaviridae (including Simian virus 40, JC virus, BK virus), Poxviridae (including vaccinia and smallpox), Sphaerolipoviridae, Stratoviridae, Turi Viridae (Turriviridae), Dinovirus (Dinodnavirus), Halotermovirus, Rhizidovirus (Rhizidovirus), Coronaviridae, Picornaviridae, Caliciviridae, Flaviviridae, Togaviridae, Bornasviridae, Filoviridae, Paramyxoviridae, Pneumoviridae, Rhabdoviridae, Arenaviridae, Bunyaviridae, Orthomyxoviridae, or Deltaviruses. Viral infections can further be caused by: Coronavirus, Poliovirus, Rhinovirus, Hepatitis A, Hepatitis B virus, Norwalk virus, Yellow fever virus, West Nile virus, Human immunodeficiency virus, Hepatitis C virus , Dengue virus, Zika virus, Rubella virus, Ross River virus, Sindbis virus, Chikungunya virus, Borna disease virus, Ebola virus, Marburg virus, Measles virus, Mumps virus, Nipah virus, Hendra virus, Newcastle disease virus, human respiratory syncytial virus, rabies virus, Lassa virus, Hantaan virus, Crimean-Congo hemorrhagic fever virus, influenza, or hepatitis D virus.

The severity of viral infection can be divided into mild, moderate, severe and critical according to clinical manifestations. Current antiviral therapies include antibody cocktails or drugs that target key processes in the viral life cycle, such as viral entry, viral uncoating, viral replication, viral synthesis, viral assembly, or viral shedding. These above-mentioned strategies tend to target specific viruses or specific classes of viruses, thus limiting their application in the face of different virus outbreaks.

Aging is associated with the progressive degeneration of tissues, which negatively affects the structure and function of vital organs, and is one of the most important known risk factors for most chronic diseases. Given that the proportion of the world's population >60 years of age is expected to double in the next four decades, the increased incidence of chronic age-related diseases will place a huge burden on healthcare resources.

Today, we are beginning to understand aging as a ubiquitous and complex phenomenon caused by environmental, stochastic, genetic and epigenetic events in different cells and tissues and their interplay throughout life. A common feature of aging tissue and age-related diseases is chronic inflammation. "Inflammatory aging" describes low-grade, chronic, systemic inflammation during aging in the absence of overt infection ("sterile" inflammation), and is a very important risk factor for morbidity and mortality in older adults. There is overwhelming epidemiological evidence that a mild inflammatory state revealed by elevated levels of inflammatory biomarkers is associated with and predicts many aging phenotypes—eg, body composition, energy production and utilization, metabolic homeostasis, immune aging and changes in neuronal health.

Therefore, targeting pathways that control age-associated inflammation across multiple systems may be beneficial in the elderly or in the treatment of age-related disorders or syndromes.

Accordingly, there is a need for new methods and compositions for the treatment and prevention of microbial infectious diseases and related inflammatory disorders. There is also a need for new methods and compositions for the treatment and prevention of aging and related diseases.

Contents of the invention

The present application provides pharmaceutical compositions and methods of using pharmaceutical combinations useful for the treatment and prevention of various aging-related disorders and syndromes, as well as for improving host responses, preventing and treating infectious diseases and their associated inflammatory disorders.

According to one aspect consistent with the principle of the present invention, a pharmaceutical composition is provided, which can be used for preventing and treating infectious diseases. According to another aspect, the pharmaceutical composition is useful for preventing and treating aging and aging-related diseases.

In some embodiments, the pharmaceutical composition may comprise therapeutic agent X or a pharmaceutically acceptable salt thereof; therapeutic agent Y or a pharmaceutically acceptable salt thereof; and at least one pharmaceutically acceptable excipient, wherein the therapeutic agent X is a non-steroidal anti-inflammatory drug and the therapeutic agent Y is a fatty acid oxidation inhibitor. The pharmaceutical compositions according to the invention are useful in methods of treating one or more conditions in a subject in need thereof, including the prevention and treatment of infectious diseases and the prevention and treatment of aging-related diseases and syndromes. The method of treating a condition in a subject in need thereof may comprise administering to the subject an effective amount of a pharmaceutical composition comprising therapeutic agent X or a pharmaceutically acceptable salt thereof; therapeutic agent Y or a pharmaceutically acceptable salt thereof; a pharmaceutically acceptable salt; and at least one pharmaceutically acceptable excipient, wherein the therapeutic agent X is a non-steroidal anti-inflammatory drug and the therapeutic agent Y is a fatty acid oxidation inhibitor.

In a further embodiment, in the pharmaceutical composition, therapeutic agent X and therapeutic agent Y are contained in a single dosage form.

In a further embodiment, in the pharmaceutical composition, therapeutic agent X and therapeutic agent Y are present in separate dosage forms.

In a further embodiment, the therapeutic agent X is selected from at least one of the following: COX inhibitors, salicylates, ibuprofen, indomethacin, flurbiprofen, phenoxy ibuprofen, Naproxen, nabumetone, piroxicam, phenylbutazone, diclofenac, fenoprofen, ketoprofen, ketorolac, tetrachlorofenoic acid, sulindac, and tometine . In some embodiments, agent X is salicylate or a derivative thereof. In some embodiments, Agent X is 2-acetyloxybenzoic acid.

In a further embodiment, the therapeutic agent Y is selected from at least one of the following: CPT inhibitors, carnitine biosynthesis inhibitors, 3-ketoacyl-CoA thiolase inhibitors, etamoxir, Hydroxyphenylglycine, perhexiline, meldonium, trimetazidine, ethoxycarnitine, aminocarnitine, or a phosphonooxy derivative of carnitine. In some embodiments, agent Y is 1-[(2,3,4-trimethoxyphenyl)methyl]piperazine.

In some embodiments, the weight ratio of therapeutic agent X to therapeutic agent Y of the pharmaceutical composition may be between 1:1 and 10:1, including 1:1, 2:1, 3:1, 4:1 , 5:1, 6:1, 7:1, 8:1, 9:1 or 10:1. In a further embodiment, the mass ratio of therapeutic agent X to therapeutic agent Y may be between 1:1 and 10:1, including 1:1, 2:1, 3:1, 4:1, 5:1 1, 6:1, 7:1, 8:1, 9:1 or 10:1.

In some embodiments, pharmaceutical compositions can be administered in oral dosage forms. In a further embodiment, the pharmaceutical composition may be administered in injectable form.

In some embodiments, therapeutic agent X and therapeutic agent Y are administered simultaneously. In some embodiments, therapeutic agent X and therapeutic agent Y are contained in a single dosage form. In some embodiments, the single pharmaceutical dosage form is an oral dosage form. In a further embodiment, the single dosage form is an injectable form.

In some embodiments, therapeutic agent X and therapeutic agent Y are administered separately. In some embodiments, therapeutic agent X is administered before therapeutic agent Y. In some embodiments, therapeutic agent X is administered after therapeutic agent Y. In some embodiments, therapeutic agent X and therapeutic agent Y are administered orally separately. In some embodiments, therapeutic agent X and therapeutic agent Y are administered separately by injection.

Description of drawings

Some embodiments of the invention are illustrated as examples and are not limited by the numbers in the figures, where like reference numerals may indicate similar elements and where:

Figure 1A – Figure 1A shows a scheme for microbial disease modeling and drug delivery.

Figure 1B - Figure 1B shows that drug combinations X and Y prevent microbial LPS-induced death in mice.

Figure 1C - Figure 1C shows drug combinations X and Y ameliorate microbial LPS-induced lung inflammation in mice.

Figure 2 - Figure 2 shows that drug combinations X and Y reduce the expression of the hepatitis C virus co-receptor CD36.

Figure 3 - Figure 3 shows that drug combinations X and Y reduce the expression of the HCV co-receptor CD81.

Figure 4 - Figure 4 shows that drug combinations X and Y reduce the expression of the hepatitis A virus co-receptor HAVCR1.

Figure 5 – Figure 5 shows that drug combinations X and Y reduce the expression of the hepatitis A virus co-receptor HAVCR2

Figure 6 - Figure 6 shows that drug combinations X and Y reduce the expression of the coronavirus co-receptor ACE2.

Figure 7 – Figure 7 shows that the drug combination X and Y reduces the expression of the coronavirus co-receptor DPP4

Figure 8 - Figure 8 shows that drug combination X and Y reduces the expression of the viral co-receptor integrin alpha(v).

Figure 9 - Figure 9 shows that drug combination X and Y reduces the expression of the viral co-receptor integrin β1.

Figure 10 - Figure 10 shows that drug combination X and Y reduces the expression of the viral co-receptor integrin β3.

Figure 11 - Figure 11 shows that drug combination X and Y reduces the expression of the viral co-receptor integrin β5.

Figure 12 - Figure 12 shows that drug combination X and Y reduces the expression of viral co-receptor integrin β6.

Figure 13A - Figure 13A shows drug combination X and Y inhibits senescence biomarkers and reduces senescent cells in aged liver tissue.

Figure 13B - Figure 13B shows that drug combinations X and Y suppress senescent biomarkers and reduce senescent cells in aged muscle tissue.

Figure 13C - Figure 13C shows drug combination X and Y inhibits aging biomarkers and reduces senescent cells in aged skin tissue.

Figure 14 - Figure 14 shows drug combinations X and Y accelerate tissue repair in aged mice with chronic wounds.

Figure 15 - Figure 15 shows drug combinations X and Y accelerate hair growth in aged mice with alopecia.

Figure 16 - Figure 16 shows that drug combinations X and Y improve tissue fibrosis in aged mice.

Figure 17 - Figure 17 shows that drug combination X and Y ameliorates inflammatory aging and inflammation-induced death in mice.

Figure 18A - Figure 18A shows drug combinations X and Y reverse inflammatory aging-associated and immunosenescence-associated gene signatures.

Figure 18B - Figure 18B shows drug combinations X and Y reverse inflammatory aging-associated and immunosenescence-associated gene signatures.

Figure 18C - Figure 18C shows drug combinations X and Y reverse inflammatory aging-associated and immunosenescence-associated gene signatures.

Figure 19A - Figure 19A shows that drug combinations X and Y promote oxidative muscle growth.

Figure 19B - Figure 19B shows that drug combinations X and Y promote oxidative muscle growth.

Figure 19C - Figure 19C shows that drug combinations X and Y promote oxidative muscle growth.

Figure 19D - Figure 19D shows that drug combinations X and Y promote oxidative muscle growth.

Figure 20 - Figure 20 shows that drug combinations X and Y induce many myokines/adipokines, just like physical exercise.

Figure 21A - Figure 21A shows that X and Y directly bind to the mitochondrial protein UQCRH according to thermal proteome analysis using the TMTpro 16plex kit and Q-Exactive HF-X mass spectrometry (Thermo Fisher).

Figure 21B - Figure 21B shows that X and Y directly bind to the mitochondrial protein NDUFS6 according to thermal proteomic analysis using the TMTpro 16plex kit and Q-Exactive HF-X mass spectrometry (Thermo Fisher).

Figure 21C - Figure 21C shows that X and Y directly bind to the mitochondrial protein COX7A2 according to thermal proteome analysis using the TMTpro 16plex kit and Q-Exactive HF-X mass spectrometry (Thermo Fisher).

Figure 22 – Figure 22 shows that X and Y synergistically reduce ATP levels within minutes.

Figure 23 - Figure 23 shows that X and Y synergistically reduce acetyl-CoA and protein acetylation.

Figure 24 - Figure 24 shows that X and Y synergistically increase Pink1 for Parkin-driven mitophagy according to immunostaining with Abcam ab23707 antibody.

Figure 25A - Figure 25A shows that X and Y synergistically increase levels of mitochondria-targeted myoblast proteins.

Figure 25B - Figure 25B shows that X and Y synergistically increase levels of mitochondria-targeted myotubes.

Figure 26 - Figure 26 shows that X and Y synergistically increase mitochondrial DNA levels according to qRT-PCR of mitochondrial genomic NADH dehydrogenases ND1 and ND4 relative to nuclear genomic B2M.

Figure 27 - Figure 27 shows that X and Y synergistically increase IREla-mediated unfolded protein response (UPR) according to RNAseq (Illumina HiSeq).

Figure 28A - Figure 28A shows that X and Y synergistically reduce mitochondrial reactive oxygen species (ROS).

Figure 28B - Figure 28B shows that X and Y synergistically reduce mitochondrial reactive oxygen species (ROS).

Figure 29A - Figure 29A shows that X and Y synergistically increase the activity of multiple cytokine pathways, including axon guidance factors (Figure 17), suggesting that drug combination X and Y delay aging by stimulating the neuroendocrine system and mimicking calorie restriction.

Figure 29B - Figure 29B shows that X and Y synergistically increase the activity of multiple cytokine pathways, including GDNF (Figure 17), suggesting that drug combination X and Y delay aging by stimulating the neuroendocrine system and mimicking calorie restriction.

Figure 29C - Figure 29C shows that X and Y synergistically increase the activity of multiple cytokine pathways, including NGF (Figure 17), suggesting that drug combination X and Y delay aging by stimulating the neuroendocrine system and mimicking calorie restriction.

Figure 30A - Figure 30A shows that drug combination X and Y synergistically increases the expression of stem cell pathways (e.g. Hippo, Notch-Hes-Hey, HIF1a and Integrin-MAPK) according to RNAseq (Hiseq), indicating increased stemness and regenerative capacity.

Figure 30B - Figure 30B shows that drug combination X and Y synergistically increase the expression of stem cell pathways (e.g. Hippo, Notch-Hes-Hey, HIF1a and Integrin-MAPK) according to RNAseq (Hiseq), indicating increased stemness and regenerative capacity.

Figure 30C - Figure 30C shows that drug combination X and Y synergistically increases the expression of stem cell pathways (e.g. Hippo, Notch-Hes-Hey, HIF1a and Integrin-MAPK) according to RNAseq (Hiseq), indicating increased stemness and regenerative capacity.

Figure 30D - Figure 30D shows that drug combination X and Y synergistically increases the expression of stem cell pathways (e.g. Hippo, Notch-Hes-Hey, HIF1a and Integrin-MAPK) according to RNAseq (Hiseq), indicating increased stemness and regenerative capacity.

Figure 30E - Figure 30E shows that drug combination X and Y synergistically increases the expression of stem cell pathways (e.g. Hippo, Notch-Hes-Hey, HIF1a and Integrin-MAPK) according to RNAseq (Hiseq), indicating increased stemness and regenerative capacity.

Figure 31 - Figure 31 shows that after 2 rounds of X and Y transdermal treatments, total body weight and waist circumference (indicators of aging-induced central obesity) in cynomolgus monkeys were both significantly reduced by about 10-20%, while calf circumference (skeletal muscle indicators of growth and regeneration) increased by about 8%.

Figure 32A - Figure 32A shows that plasma levels of the neuroendocrine cytokine BDNF (R&D Systems ELISA) in cynomolgus monkeys were also significantly increased after treatment with the drug combination X and Y, but either X alone or Y alone or vehicle Does not have this effect.

Figure 32B - Figure 32B shows that plasma levels of the neuroendocrine cytokine LIF (R&D Systems ELISA) in cynomolgus monkeys were also significantly increased following treatment with the drug combination X and Y, but X alone or Y alone or vehicle had no such effect .

Figure 33A - Figure 33A shows that no toxicity related to drug combination X and Y was observed based on clinical hematology or clinical blood chemistry (Figure 21 ).

Figure 33B - Figure 33B shows that no toxicity related to drug combination X and Y was observed based on clinical hematology or clinical blood chemistry (Figure 21 ).

Figure 34 – Figure 34 shows that X and Y promote liver regeneration in 1-year-old diabetic mice with age-associated steatohepatitis, as Ki67 (ab16667) relative to X alone or Y alone or vehicle Increased proliferation of hepatocytes indicated by immunostaining was observed.

Figure 35 - Figure 35 shows that drug combination X and Y reduces beta islet fibrosis in the pancreas of age-associated pancreatitis due to type 2 diabetes according to Masson's trichrome staining.

Figure 36A - Figure 36A shows that drug combinations X and Y increase the growth and proliferation of beta islet cells according to Ki67 (Ab16667) immunostaining

Figure 36B - Figure 36B shows that drug combination X and Y increase the growth and proliferation of beta islet cells according to Ki67 (Ab16667) immunostaining

Figure 37A - Figure 37A shows that drug combination X and Y increases beta islet cell growth and proliferation, thereby promoting islet beta islet regeneration according to qRT-PCR using primer pairs Pax6, Mafa and Pdx1 from Origene.

Figure 37B - Figure 37B shows that drug combination X and Y increases beta islet cell growth and proliferation, thereby promoting islet beta islet regeneration according to qRT-PCR using primer pair Ins1 and Ins2 from Origene.

Figure 38 - Figure 38 shows that drug combination X and Y promote senescence hair regrowth and wound healing by 64 days.

Figure 39 - Figure 39 shows that X and Y increase protein levels of myosin heavy chain (MHC; Sigma-Aldrich-M4276 antibody) in the quadriceps muscle of 1-year-old diabetic mice, thereby promoting skeletal muscle regeneration.

Figure 40 - Figure 40 shows that drug combination X and Y significantly increased total body weight compared to aged control animals with muscle wasting and sarcopenia, indicating that X and Y reversed sarcopenia.

Figure 41 - Figure 41 shows that daily oral gavage of the drug combination X and Y over 3 weeks resulted in a significant increase in tracheal bleomycin (3mg/kg in 50ul phosphate) compared to either X alone or Y alone or vehicle. Aging-associated pulmonary fibrosis was reduced in 1-year-old mice treated with saline-buffered saline or PBS) to mimic chronic obstructive pulmonary disease (COPD) and idiopathic pulmonary fibrosis (IPF).

Figure 42 - Figure 42 shows that drug combinations X and Y restore the estrous cycle in most female rats (PCOS rat model) based on vaginal smear analysis.

Detailed ways

The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. As used herein, the term "and/or" includes any and all combinations of one or more of the associated listed items. As used herein, the singular forms "a, an" and "the" are intended to include plural as well as singular forms unless the context clearly dictates otherwise. It will be further understood that when used in this specification, the terms "comprises and/or comprising" indicate the presence of stated features, steps, operations, elements, and/or components, but do not exclude The presence or addition of one or more other features, steps, operations, elements, components, and/or groups thereof.

Unless otherwise defined, all terms (including technical and scientific terms) used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. It will be further understood that terms such as those defined in commonly used dictionaries should be interpreted as meanings consistent with their meanings in relevant technical field documents and this disclosure, and will not be interpreted in an idealized or overly formal sense, except in This is clearly defined.

The term "microorganism" refers to microscopic organisms including viruses, bacteria, fungi, protozoa, algae, amoebae, and slime molds, as well as other potentially pathogenic organisms.

The term "infection" refers to the invasion of the body tissues of an organism by pathogenic agents, their reproduction, and the response of host tissues to the infectious agents and the toxins they produce.

The term "aging-related disease" refers to diseases and degenerative diseases whose incidence increases with age.

The term "inflammatory aging" refers to the invasion of pathogenic agents into the body tissues of an organism, their reproduction, and the response of host tissues to infectious agents and the toxins they produce.

The term "NSAID" or "nonsteroidal anti-inflammatory drug" is used interchangeably herein to refer to a member of a class of drugs that inhibit cyclooxygenase (COX) activity to relieve pain, reduce fever, and prevent drug clots. It is known in the art that not all inflammatory disorders can be treated with NSAIDs.

The terms "fatty acid oxidation" and "FAO" are used interchangeably herein to refer to the biochemical process of breaking down fatty acids into acetyl-CoA units. In some embodiments, the FAO is located in the mitochondria of the cell. In some embodiments, the FAO is in the peroxisomes of the cell. In some embodiments, the FAO rate-limiting step is catalyzed by carnitine palmitoyltransferase (CPT).

As used herein, "treatment" or "treating" is a method used to obtain beneficial or desired results, including clinical results. For purposes of the present invention, beneficial or desired clinical outcomes include, but are not limited to, one or more of the following: reduction of one or more symptoms caused by the disease, reduction of the extent of the disease, stabilization of the disease or condition ( For example, to prevent or delay the progression of a disease or condition), to prevent or delay the spread of a disease or condition, to prevent or delay the onset or recurrence of a disease or condition, to delay or slow the progression of a disease or condition, to ameliorate a disease state, to provide Remission (whether partial or total), reducing the dose of one or more other drugs needed to treat a disease or condition, delaying the progression of a disease or condition, improving quality of life and/or prolonging survival. "Treatment" also includes alleviation of the pathological consequences of a disease or condition. The methods of the present application utilize any one or more of these therapeutic aspects.

As used herein, the terms "prevention and prophylaxis" refer to measures used to prevent or prevent a disease, disorder or condition from occurring in a subject at risk of disease but not yet diagnosed. Prophylaxis (and an effective prophylactic dose) can be demonstrated in population studies, e.g., an amount effective to prevent or prevent a given disease or medical condition is effective in reducing morbidity in treated subjects relative to untreated control subjects. method.

The terms "individual", "subject" and "patient" are used interchangeably herein to describe mammals, including humans. A subject includes, but is not limited to, a human, bovine, ovine, porcine, equine, feline, canine, rodent, or primate. In some embodiments, the individual is a human. In some embodiments, the individual has a disease or condition. In some embodiments, the individual is in need of treatment.

As understood in the art, an "effective amount" refers to an amount of a composition sufficient to produce a desired therapeutic outcome. For therapeutic use, beneficial or desired results include, for example, reduction of one or more symptoms (biochemical, histological, and/or behavioral) caused by a disease or disorder, including complications thereof that arise during the development of the disease or disorder and intermediate pathological phenotypes, increasing the quality of life of those individuals with the disease or condition, reducing the dose of other drugs needed to treat the disease or condition, enhancing the effect of another drug, delaying the progression of the disease or condition, and/or prolong the survival of patients.

The term "pharmaceutically acceptable salt" means a salt (a salt with a counterion having acceptable mammalian safety for a given dosage regimen) that is acceptable for administration to a patient, such as a mammal. Such salts can be derived from pharmaceutically acceptable inorganic or organic bases, and pharmaceutically acceptable inorganic or organic acids. "Pharmaceutically acceptable salt" refers to a pharmaceutically acceptable salt of a compound derived from various organic and inorganic counterions as known in the art, and by way of example only, sodium, potassium, calcium, magnesium, Ammonium, tetraalkyl, etc.; and when the molecule contains basic functionality, salts of organic or inorganic acids such as hydrochloride, hydrobromide, formate, tartrate, benzenesulfonate, methanesulfonate , acetate, maleate, oxalate, etc.

The term "salt thereof" refers to a compound formed when a proton of an acid is replaced by a cation such as a metal cation or an organic cation or the like. Where applicable, the salt is a pharmaceutically acceptable salt, although this is not essential for salts of intermediate compounds which are not intended to be administered to a patient. For example, salts of compounds of the invention include those wherein the compound is protonated with an inorganic or organic acid to form a cation and the conjugate base of the inorganic or organic acid is the anionic component of the salt.

The terms "X and Y" or "X+Y" or "XY" are used interchangeably herein to refer to a compound comprising therapeutic agent X or a pharmaceutically acceptable salt thereof and therapeutic agent Y or a pharmaceutically acceptable salt thereof. Salt combination.

Therapeutic agent X or a pharmaceutically acceptable salt thereof and therapeutic agent Y may be administered together or sequentially.

In describing the present invention, it is to be understood that numerous techniques and steps are disclosed. Each of these has its own benefits and each can also be used in combination with one or more, or in some cases all, of the other disclosed techniques. Therefore, for the sake of clarity, this description will avoid unnecessarily repeating every possible combination of individual steps. However, the description and claims should be understood as such combinations are fully within the scope of the invention and claims.

As used in this application, the term "about" or "approximately" refers to a range of values within plus or minus 20% of the indicated figure. Furthermore, as used in this application, the term "substantially" means that the actual value is within about 10% of the actual expected value, especially within about 5% of the actual expected value, especially with respect to any variables, elements or limitations specified herein. Within about 1% of the actual expected value.

Novel methods and compositions for the treatment and prevention of aging and related diseases and for the treatment and prevention of microbial infectious diseases and related inflammatory disorders are discussed herein. In the following description, for purposes of explanation, numerous specific details are set forth in order to provide a thorough understanding of the present invention. It will be apparent, however, to one skilled in the art that the present invention may be practiced without these specific details.

This disclosure is to be considered as an example of the invention and is not intended to limit the invention to the specific embodiments illustrated in the following figures or description.

The invention will now be described by way of example and with reference to the accompanying drawings representing preferred and alternative embodiments. According to an embodiment of the present invention, there is provided a composition of matter useful for treating various diseases, conditions and infections. In some embodiments, the composition of matter may include a pharmaceutical composition having: therapeutic agent X or a pharmaceutically acceptable salt thereof; therapeutic agent Y or a pharmaceutically acceptable salt thereof; and at least one pharmaceutical An acceptable excipient wherein the therapeutic agent X is a non-steroidal anti-inflammatory drug and the therapeutic agent Y is a fatty acid oxidation inhibitor. In some embodiments, therapeutic agent X is salicylic acid or a derivative thereof (eg, salicylic acid). In some embodiments, therapeutic agent X is 2-acetyloxybenzoic acid. In some embodiments, the therapeutic agent Y is 1-[(2,3,4-trimethoxyphenyl)methyl]piperazine. According to another embodiment of the present invention, there is provided a method of treating one or more conditions, including the prevention and treatment of infectious diseases and the prevention and treatment of aging-related diseases and syndromes, in a subject in need thereof. treat. The method of treating a condition in a subject in need thereof may comprise administering to the subject an effective amount of a pharmaceutical composition comprising therapeutic agent X or a pharmaceutically acceptable salt thereof; therapeutic agent Y or a pharmaceutically acceptable salt thereof; a pharmaceutically acceptable salt; and at least one pharmaceutically acceptable excipient, wherein the therapeutic agent X is a non-steroidal anti-inflammatory drug and the therapeutic agent Y is a fatty acid oxidation inhibitor.

Pharmaceutical compositions and methods comprising administering pharmaceutical compositions according to the invention are effective in treating or preventing microbial infections.

In some embodiments, the pharmaceutical composition comprising therapeutic agent X or a pharmaceutically acceptable salt thereof and therapeutic agent Y or a pharmaceutically acceptable salt thereof can effectively prevent and/or treat viral infection by modulating the immune system . In some preferred embodiments, the pharmaceutical composition comprising therapeutic agent X or a pharmaceutically acceptable salt thereof and therapeutic agent Y or a pharmaceutically acceptable salt thereof can regulate immune regulatory factors, such as but not limited to: ACVR1 , ACVR1B, ACVR2A, ACVR2B, ACVRL1, AMH, AMHR2, BMP2, BMP7, BMPR1A, BMPR1B, BMPR2, CCL11, CCL15, CCL17, CCL19, CCL2, CCL20, CCL21, CCL22, CCL23, CCL24, CCL27, CCL28, CCL3, CCL4 , CCL5, CCL7, CCL8, CCR1, CCR10, CCR2, CCR3, CCR4, CCR5, CCR6, CCR7, CCR8, CCR9, CD27, CD40, CD40LG, CD70, CLCF1, CNTF, CNTFR, CRLF2, CSF1, CSF1R, CSF2, CSF2RA , CSF2RB, CSF3, CSF3R, CTF1, CX3CL1, CX3CR1, CXCL10, CXCL11, CXCL12, CXCL13, CXCL14, CXCL16, CXCL6, CXCL9, CXCR1, CXCR2, CXCR3, CXCR4, CXCR5, CXCR6, EDA, EDA2R, EDAR, EGF , EGFR , EPO, EPOR, FAS, FASLG, FLT1, FLT3, FLT3LG, FLT4, GDF5, GH1, GHR, HGF, IFNA1, IFNA10, IFNA17, IFNA2, IFNA4, IFNA5, IFNA6, IFNA7, IFNA8, IFNAR1, IFNAR2, IFNB1, IFNE , IFNG, IFNGR1, IFNGR2, IFNK, IFNL3, IFNLR1, IL10, IL10RA, IL10RB, IL11, IL11RA, IL12A, IL12B, IL12RB1, IL12RB2, IL13, IL13RA1, IL15, IL15RA, IL17A, IL17B, IL17RA, IL17RB, IL18, IL18R1 , IL18RAP, IL19, IL1A, IL1B, IL1R1, IL1R2, IL1RAP, IL2, IL20, IL20RA, IL20RB, IL21, IL21R, IL22, IL22RA1, IL22RA2, IL23A, IL23R, IL24, IL25, IL2RA, IL2RB, IL2RG, IL3, IL4 , IL4R, IL5, IL5RA, IL6, IL6R, IL6ST, IL7, IL7R, IL9, IL9R, INHBA, INHBB, INHBC, INHBE, KDR, KIT, KITLG, LEP, LEPR, LIF, LIFR, LTA, LTB, MET, MPL , NGFR, OSM, OSMR, PDGFA, PDGFB, PDGFC, PDGFRA, PDGFRB, PLEKHO2, PPBP, PRL, PRLR, RELT, TGFB1, TGFB2, TGFB3, TGFBR1, TGFBR2, TNFA, TNFRSF10B, TNFRSF10C, TNFRSF10D, TNFRSF11A, TNFRSF11B, TNF RSF12A , TNFRSF13B, TNFRSF13C, TNFRSF14, TNFRSF17, TNFRSF18, TNFRSF19, TNFRSF1A, TNFRSF1B, TNFRSF21, TNFRSF25, TNFRSF4, TNFRSF8, TNFRSF9, TNFSF10, TNFSF11, TNFSF12, TNFSF13, TN FSF13B, TNFSF14, TNFSF15, TNFSF18, TNFSF4, TNFSF8, TNFSF9, TPO , TSLP, VEGFA, VEGFB, VEGFC, VEGFD, XCL1, XCR1. In another aspect, the present invention provides a method of treating a condition in a subject in need thereof by modulating one or more of the above-mentioned immunomodulatory factors, the method comprising administering to the subject an effective amount of a pharmaceutical composition.

In some embodiments, a pharmaceutical composition comprising therapeutic agent X or a pharmaceutically acceptable salt thereof and therapeutic agent Y or a pharmaceutically acceptable salt thereof can effectively prevent and/or treat viral infection by modulating inflammation. In some preferred embodiments, the pharmaceutical composition comprising therapeutic agent X or a pharmaceutically acceptable salt thereof and therapeutic agent Y or a pharmaceutically acceptable salt thereof can reduce the expression of factors, such as but not limited to: AKT1 , AKT2, AKT3, BTK, CASP8, CCL3, CCL4, CCL5, CD14, CD180, CD40, CD80, CD86, CHUK, CISH, CTNNAL1, CUEDC2, CXCL10, CXCL11, CXCL8, CXCL9, CYLD, FADD, FBXW5, FOS, GM -CSF, IFNA1, IFNA10, IFNA13, IFNA14, IFNA16, IFNA17, IFNA2, IFNA21, IFNA4, IFNA5, IFNA6, IFNA7, IFNA8, IFNAR1, IFNAR2, IFNB1, IFNG, IKBKB, IKBKE, IKBKG, IL12A, IL12B, IL18, IL1B , IL6, IL8, IRAK1, IRAK2, IRAK3, IRAK4, IRF3, IRF5, IRF7, JUN, LBP, LY96, MAP2K1, MAP2K2, MAP2K3, MAP2K4, MAP2K6, MAP2K7, MAP3K7, MAP3K8, MAPK1, MAPK10, MAPK11, MAPK12, MAPK1 3 , MAPK14, MAPK3, MAPK8, MAPK9, MBL2, MIR105-1, MIR6502, MIR718, MIR98, MIRLET7E, MIRLET7I, MLST8, MYD88, NFKB1, NFKB2, NFKBIA, OTUD5, PELI1, PELI2, PELI3, PIK3CA, PIK3CB, PIK3CD, PIK3CG , PIK3R1, PIK3R2, PIK3R3, PIK3R5, PLK1, PROBE, PTPN6, RAC1, RBCK1, RELA, RIPK1, RNF216, RNF31, RNF41, SARM1, SFTPA2, SFTPD, SIGIRR, SMAD6, SOCS1, SPP1, SQSTM1, STAT1, SYK, TAB1 , TAB2, TAB3, TBK1, TICAM1, TICAM2, TIFA, TIRAP, TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8, TLR9, TMED7, TNFA, TNFAIP3, TOLLIP, TRAF3, TRAF6, TRAFD1, TREM1, USP7 , ZMYND11. In another aspect, the present invention provides a method of treating a condition in a subject in need thereof by reducing the expression of one or more of the aforementioned factors, the method comprising administering to the subject an effective amount of a pharmaceutical composition.

In some embodiments, a pharmaceutical composition comprising therapeutic agent X or a pharmaceutically acceptable salt thereof and therapeutic agent Y or a pharmaceutically acceptable salt thereof can effectively prevent and/or treat viruses by reducing viral receptors Infect. In some preferred embodiments, the pharmaceutical composition comprising therapeutic agent X or a pharmaceutically acceptable salt thereof and therapeutic agent Y or a pharmaceutically acceptable salt thereof can reduce the expression of factors, such as but not limited to: ACE2 , CD4, CD36, CD81, CLDN1, CXCR4, DPP4, HAVCR1, HAVCR2, ITGAV, ITGB1, ITGB3, ITGB5, ITGB6, MAG, SLC1A4, SLC1A5, SLC7A1, SLC20A1, SLC20A2. In another aspect, the present invention provides a method of treating a condition in a subject in need thereof by reducing the expression of one or more of the aforementioned factors, the method comprising administering to the subject an effective amount of a pharmaceutical composition.

In some embodiments, the pharmaceutical composition comprising therapeutic agent X or a pharmaceutically acceptable salt thereof and therapeutic agent Y or a pharmaceutically acceptable salt thereof can effectively prevent and or treat diseases caused by viruses, bacteria, fungi, Microbial infections caused by protozoa, or parasites. In a further embodiment, a method of effectively preventing and/or treating microbial infection caused by a virus, bacterium, fungus, protozoa or parasite may comprise administering to the subject an effective amount of a pharmaceutical composition.

In the case of a viral infection, it may be caused by a DNA virus. In some embodiments, DNA viruses may include, but are not necessarily limited to, members of the following: Adenoviridae, Papovaviridae, Parvoviridae, Anoroviridae, Pleolipoviridae, Myoviruses family, short-tailed bacteriophages, long-tailed bacteriophages, heteroherpesviridae, herpesviridae (including human herpesviruses, and varicella-zoster virus), Malocoherpesviridae (Malocoherpesviridae), fat hair bacteriophages, ancient bacteriophages Family, Adenoviridae, Ampullaviridae, Vesiviridae, African Swine Fever Viridae (including ASFV), Baculoviridae, Sikandaviridae, Rhabdoviridae, Covering Phage Family , small spindle-shaped bacteriophage family, spheroviridae, titriviridae, mast sialoviridae, iridoviridae, Maseilleviridae (Maseilleviridae), mimiviridae, nudeviridae, linear virus family (Nimaviridae), Pandoraviridae, Papillomaviridae, Phytoviridae, Blastoviridae, Polydnaviruses, Polyomaviridae (including Simian Virus 40, JC Virus, BK Virus), Poxviruses family (including vaccinia and smallpox), globosaviridae, tomiviridae, tuuriviridae, Dinovirus, halotermovirus, or Ryzevirus.

Viral infections can also be caused by double-stranded RNA viruses, positive-sense RNA viruses, negative-sense RNA viruses, retroviruses, or combinations thereof. Viral infections can further be caused by: Coronaviridae, Picornaviridae, Reoviridae, Caliciviridae, Flaviviridae, Togaviridae, Bornaviridae, Silk Viridae, Paramyxoviridae, Pneumoviridae, Rhabdoviridae, Arenaviridae, Bunyaviridae, Astroviridae Orthomyxoviridae, Arteriviridae , hepatitis virus, retroviridae virus, cauliviridae virus, hepadnaviridae virus, or delta virus. Viral infections can further be caused by: Coronavirus, Poliovirus, Rhinovirus, Hepatitis A, Norwalk Virus, Yellow Fever Virus, West Nile Virus, Hepatitis C Virus, Dengue Virus, Zika Virus, Rubella Virus , Ross River virus, Sindbys virus, Chikungunya virus, Borna disease virus, Ebola virus, Marburg virus, measles virus, mumps virus, Nigeria Pavirus, Hendra virus, Newcastle disease virus, human respiratory syncytial virus, rabies virus, Lassa virus, Hantaan virus, Crimean-Congo hemorrhagic fever virus, influenza, or hepatitis D virus. In some preferred embodiments, the viral infection is caused by a coronavirus. Examples of coronaviruses include, but are not limited to, alphacoronavirus 1, human coronavirus 229E, human coronavirus NL63, longwing bat coronavirus 1, longwing bat coronavirus HKU8, porcine epidemic diarrhea virus, chrysanthemum Bat-coronavirus HKU2, Bat-coronavirus 512, Betacoronavirus 1 (bovine coronavirus, human coronavirus OC43), hedgehog-coronavirus 1, human coronavirus HKU1, Middle East Respiratory Syndrome-related coronavirus Virus, murine coronavirus, bat coronavirus HKU5 of the genus Drosophila, bat coronavirus HKU9 of the genus Drosophila, severe acute respiratory syndrome-associated coronavirus (SARS-CoV, SARS-CoV-2), bat coronavirus HKU4, avian coronavirus, beluga coronavirus SW1, nightingale coronavirus HKU11 and porcine coronavirus HKU15. In some preferred embodiments, the viral infection is caused by SARS-CoV2.

In other embodiments, the infection can be bacterial in nature. Bacteria causing bacterial infections may include, but are not necessarily limited to, Acinetobacter species, Actinomycetes species, Actinomyces species, Aeromonas species, Aeromonas species species, Rhodospora spp., Alcaligenes spp., Bacillus spp., Bacteroides spp., Bartonella spp., Bifidobacterium spp., Bordetella spp., Borrelia spp., Brucella spp. Chlamydia sp., Burkholderia sp., Campylobacter sp., Capnocytophaga sp., Chlamydia sp., Citrobacter sp., Coxiella sp., Corynebacterium sp., Clostridium sp. , Eikenella spp., Enterobacter spp., Escherichia spp., Enterococcus spp., Ehlichia spp., Epidermophyton spp., Erysipelothrix spp., Eubacterium spp. species, Francisella spp., Fusobacterium spp., Gardnerella spp., Geminococcus spp., Haemophilus spp., Helicobacter spp., Kingella spp., Klebsiella spp., milk Bacillus spp., Lactococcus spp., Listeria spp., Leptospira spp., Legionella spp., Leptospira spp., Leuconostoc spp., Mansonella spp., Microsporum spp., Micrococcus spp., Moraxella spp., Morganella spp., Kinesiella spp., Micrococcus spp., Mycobacterium spp., Mycoplasma spp., Nocardia spp., Neisseria spp., Pasteurelaa spp., Pediococcus spp., Peptostreptococcus spp., Pityrophyton spp., Pseudomonas spp., Prevotella spp., Porphyromonas spp., Proteus spp., Proteus spp. Widensella sp., Pseudomonas sp., Propionibacterium sp., Rhodococcus sp., Rickettsia sp., Rhodococcus sp., Serratia sp., Stenotrophomonas sp. , Salmonella spp., Serratia spp., Shigella spp., Staphylococcus spp., Streptococcus spp., Spirillum spp., Streptobacter spp., Treponema spp., Tropheryma spp. species, Trichophyton spp., Ureaplasma spp., Veillonella spp., Vibrio spp., Yersinia spp., Xanthomonas spp., or combinations thereof.

In other embodiments, the infection may be fungal and may be caused by fungi such as, but not necessarily limited to Aspergillus, Blastomyces, Candidiasis, Coccidioidomycosis, Cryptococcus neoformans, Cryptococcus gatti ), species Histoplasma spp. (e.g. Histoplasma capsulatus), Pneumocystis spp. (e.g. Pneumocystis jirovecii), Staphylococcus sp. (e.g. Staphylococcus nigeri), Mucroymcosis ), Sporotrichum sp., Fungal ocular infectious tinea, Umbilium sp., Cladomyces sp., Geotrichum sp., Saccharomyces sp., Hansenula sp., Candida sp., Kluyveromyces sp. , Debariae spp., Pichia spp., Penicillium spp., Cladosporium spp., Thylacia spp., or combinations thereof.

In other embodiments, the infection may be caused by protozoa, such as Euglenozoa, Heterolobosea, Diplomonadida, Amoebozoa, Blastocystic, Apicomplexa ) or a combination thereof.

In other embodiments, the infection may be caused by a parasite such as, but not necessarily limited to, Trypanosoma cruzi (Chagas disease), Trypanosoma brucei gambiae, Trypanosoma brucei Rhodesia, Leishmania brasiliensis , Leishmania infantum, Leishmania mexicana, Leishmania major (L.major), Leishmania tropicalis, Leishmania donovani, Naegleria fowleri, Giardia intestinalis ( Lambertia, Giardia duodenum), canthamoebacastellanii, Balamuthiamadrillaris, Dysentery amoebae, Blastocystichominis, Babesia parvum, Cryptosporidium, Cyclosporidium, Plasmodium falciparum, Interstitium Plasmodium vivax, Plasmodium ovale, Plasmodium malariae, and Toxoplasma gondii, or a combination thereof.

In some embodiments, pharmaceutical compositions and methods comprising administering pharmaceutical compositions according to the invention are capable of treating and/or preventing related inflammatory disorders associated with cytokine and immune dysfunction, such as but not limited to CRS, ARDS, ALI , AIDS, asthma, chronic peptic ulcer, tuberculosis, rheumatoid arthritis, periodontitis, ulcerative colitis, Crohn's disease, sinusitis, encephalitis, myelitis, meningitis, arachnoiditis, neuritis, Lacrimal gland inflammation, scleritis, episcleritis, keratitis, retinitis, chorioretinitis, blepharitis, conjunctivitis, uveitis, otitis externa, otitis media, labyrinthitis, mastoiditis, carditis, endocardium Inflammation, myocarditis, pericarditis, vasculitis, arteritis, phlebitis, capillary vasculitis, sinusitis, rhinitis, pharyngitis, laryngitis, tracheitis, bronchitis, bronchiolitis, pneumonia, pleurisy, mediastinitis, stomatitis, Gingivitis, gingivostomatitis, glossitis, tonsillitis, sialadenitis, parotitis, cheilitis, pulpitis, jaw inflammation, esophagitis, gastritis, gastroenteritis, enteritis, colitis, enterocolitis, twelve Enteritis, ileitis, appenditis, appendicitis, proctitis, hepatitis, ascending cholangitis, cholecystitis, pancreatitis, peritonitis, dermatitis, folliculitis, cellulitis, hidradenitis, arthritis, dermatomyositis, myositis, Synovitis, tenosynovitis, bursitis, enthesitis, fasciitis, bursitis, epicondylitis, tendonitis, panniculitis, osteochondritis, osteitis, osteomyelitis, spondylitis, periostitis, chondritis , nephritis, glomerulonephritis, pyelonephritis, ureteritis, cystitis, urethritis, oophoritis, salpingitis, endometritis, parauterine tissue inflammation, cervicitis, vaginitis, vulvitis, mastitis, orchitis , epididymitis, prostatitis, seminal vesiculitis, balanitis, posthitis, balanoposthitis, chorioamnionitis, umbilical cord inflammation, omphalitis, insulitis, hypophysitis, thyroiditis, parathyroiditis, adrenalitis, lymphatic vessels inflammation or lymphadenitis.

In some embodiments, pharmaceutical compositions and methods comprising administering a pharmaceutical composition according to the invention are capable of treating and/or preventing aging-related diseases, the pharmaceutical composition comprising a therapeutic agent X or a pharmaceutically acceptable salt thereof and treating Sexual agent Y or a pharmaceutically acceptable salt thereof.

In some embodiments, pharmaceutical compositions and methods comprising therapeutic agent X or a pharmaceutically acceptable salt thereof and therapeutic agent Y or a pharmaceutically acceptable salt thereof are effective in preventing and/or treating aging by promoting regeneration related diseases.

In some embodiments, pharmaceutical compositions and methods comprising therapeutic agent X or a pharmaceutically acceptable salt thereof and therapeutic agent Y or a pharmaceutically acceptable salt thereof are effective in preventing and or treating aging by reversing fibrosis related diseases.

In some embodiments, pharmaceutical compositions and methods comprising therapeutic agent X or a pharmaceutically acceptable salt thereof and therapeutic agent Y or a pharmaceutically acceptable salt thereof are effective in preventing and or treating aging-related disease. The effects of therapeutic agent X and therapeutic agent Y can be assessed by testing the therapeutic agents against relevant cell and tissue models, some of which are detailed in the listed examples. Alternatively, the effect of therapeutic agent X and therapeutic agent Y can be assessed by testing the therapeutic agents against relevant animal models, some of which are described in the Examples.

In some embodiments, pharmaceutical compositions and methods comprising therapeutic agent X or a pharmaceutically acceptable salt thereof and therapeutic agent Y or a pharmaceutically acceptable salt thereof can improve insulin sensitivity or prevent or treat insulin resistance. effectively prevent and or treat age-related diseases. In some preferred embodiments, the pharmaceutical composition comprising therapeutic agent X or a pharmaceutically acceptable salt thereof and therapeutic agent Y or a pharmaceutically acceptable salt thereof can improve glucose tolerance by improving insulin sensitivity.

In some embodiments, pharmaceutical compositions and methods comprising therapeutic agent X or a pharmaceutically acceptable salt thereof and therapeutic agent Y or a pharmaceutically acceptable salt thereof are effective in preventing and or treating aging and Aging-related diseases. In some embodiments, the pharmaceutical composition comprising therapeutic agent X or a pharmaceutically acceptable salt thereof and therapeutic agent Y or a pharmaceutically acceptable salt thereof can effectively prevent or treat or reduce chronic inflammation or inflammatory aging Prevention and or treatment of aging and aging-related diseases. In some preferred embodiments, pharmaceutical compositions and methods comprising therapeutic agent X or a pharmaceutically acceptable salt thereof and therapeutic agent Y or a pharmaceutically acceptable salt thereof can reduce the expression of factors such as but not limited to : AKT1, AKT2, AKT3, BTK, CASP8, CCL3, CCL4, CCL5, CD14, CD180, CD40, CD80, CD86, CHUK, CISH, CTNNAL1, CUEDC2, CXCL10, CXCL11, CXCL8, CXCL9, CYLD, FADD, FBXW5, FOS , GM-CSF, IFNA1, IFNA10, IFNA13, IFNA14, IFNA16, IFNA17, IFNA2, IFNA21, IFNA4, IFNA5, IFNA6, IFNA7, IFNA8, IFNAR1, IFNAR2, IFNB1, IFNG, IKBKB, IKBKE, IKBKG, IL12A, IL12B, IL18 , IL1B, IL6, IL8, IRAK1, IRAK2, IRAK3, IRAK4, IRF3, IRF5, IRF7, JUN, LBP, LY96, MAP2K1, MAP2K2, MAP2K3, MAP2K4, MAP2K6, MAP2K7, MAP3K7, MAP3K8, MAPK1, MAPK10, MAPK11, MAPK12 , MAPK13, MAPK14, MAPK3, MAPK8, MAPK9, MBL2, MIR105-1, MIR6502, MIR718, MIR98, MIRLET7E, MIRLET7I, MLST8, MYD88, NFKB1, NFKB2, NFKBIA, OTUD5, PELI1, PELI2, PELI3, PIK3CA, PIK3CB , PIK3CD , PIK3CG, PIK3R1, PIK3R2, PIK3R3, PIK3R5, PLK1, PROBE, PTPN6, RAC1, RBCK1, RELA, RIPK1, RNF216, RNF31, RNF41, SARM1, SFTPA2, SFTPD, SIGIRR, SMAD6, SOCS1, SPP1, SQSTM1, STAT1, SYK , TAB1, TAB2, TAB3, TBK1, TICAM1, TICAM2, TIFA, TIRAP, TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8, TLR9, TMED7, TNFA, TNFAIP3, TOLLIP, TRAF3, TRAF6, TRAFD1, TREM1 , USP7, ZMYND11.

In some embodiments, pharmaceutical compositions and methods comprising therapeutic agent X or a pharmaceutically acceptable salt thereof and therapeutic agent Y or a pharmaceutically acceptable salt thereof are effective in preventing and or treating aging-related diseases, such as Alzheimer's disease, Parkinson's disease, dementia atherosclerosis, cardiovascular disease, type II diabetes, cancer, arthritis, rheumatoid arthritis, periodontitis, cataract, osteoporosis, fibrosis, cirrhosis , idiopathic pulmonary fibrosis, cardiac fibrosis, uterine fibrosis, scarring, joint fibrosis, chronic kidney disease, Crohn's disease, keloids, myelofibrosis, retroperitoneal fibrosis, scleroderma, sclerosis, Chronic wounds (eg, diabetic foot ulcers), chronic skin fibrosis, skin fibrosis, skin aging, nonalcoholic steatohepatitis (NASH), hair loss, tissue atrophy, menopause, primary ovarian insufficiency, endometrial hyperplasia , adenomyosis and sarcopenia.

In some preferred embodiments, the pharmaceutical composition and method comprising therapeutic agent X or a pharmaceutically acceptable salt thereof and therapeutic agent Y or a pharmaceutically acceptable salt thereof can effectively prevent and or treat fibrosis. It is widely accepted in the art that fibrosis, inflammation-induced fibrosis, inflammation and inflammatory aging can be simulated in animals, for example by administering bleomycin or liposomes to animal models (e.g., mice or rats). polysaccharides.

In some preferred embodiments, the pharmaceutical composition and method comprising therapeutic agent X or a pharmaceutically acceptable salt thereof and therapeutic agent Y or a pharmaceutically acceptable salt thereof can effectively prevent and or treat pulmonary fibrosis . Non-limiting examples of pulmonary fibrosis include, but are not limited to, chronic obstructive pulmonary disease (COPD), idiopathic pulmonary fibrosis (IPF), interstitial lung disease (ILD).

In some preferred embodiments, the pharmaceutical composition and method comprising therapeutic agent X or a pharmaceutically acceptable salt thereof and therapeutic agent Y or a pharmaceutically acceptable salt thereof can effectively prevent and or treat hair loss or hair loss. Non-limiting examples of hair loss include, but are not limited to, alopecia areata, alopecia totalis, and alopecia universalis, and androgenetic alopecia.

In some preferred embodiments, the pharmaceutical composition and method comprising therapeutic agent X or a pharmaceutically acceptable salt thereof and therapeutic agent Y or a pharmaceutically acceptable salt thereof can effectively prevent and or treat liver cirrhosis.

In some preferred embodiments, the pharmaceutical composition and method comprising therapeutic agent X or a pharmaceutically acceptable salt thereof and therapeutic agent Y or a pharmaceutically acceptable salt thereof can effectively prevent and or treat NASH.

In some preferred embodiments, the pharmaceutical composition and method comprising therapeutic agent X or a pharmaceutically acceptable salt thereof and therapeutic agent Y or a pharmaceutically acceptable salt thereof can effectively prevent and or treat type II diabetes .

In some preferred embodiments, the pharmaceutical composition and method comprising therapeutic agent X or a pharmaceutically acceptable salt thereof and therapeutic agent Y or a pharmaceutically acceptable salt thereof can effectively prevent and or treat chronic wounds.

In some preferred embodiments, the pharmaceutical composition and method comprising therapeutic agent X or a pharmaceutically acceptable salt thereof and therapeutic agent Y or a pharmaceutically acceptable salt thereof can effectively prevent and/or treat female-specific Aging-related diseases such as but not limited to: menopause, primary ovarian insufficiency, endometrial hyperplasia, adenomyosis, polycystic ovary syndrome.

In some preferred embodiments, pharmaceutical compositions and methods comprising therapeutic agent X or a pharmaceutically acceptable salt thereof and therapeutic agent Y or a pharmaceutically acceptable salt thereof are effective in preventing and/or treating sarcopenia disease.

In some preferred embodiments, the pharmaceutical composition and method comprising therapeutic agent X or a pharmaceutically acceptable salt thereof and therapeutic agent Y or a pharmaceutically acceptable salt thereof can effectively prevent and or treat sarcopenia . Sarcopenia, a disorder characterized by loss of skeletal muscle mass and function, is highly prevalent in older adults, especially older men. Therefore, an increase in muscle mass can be seen as a way to improve or treat this condition.

In some embodiments, the present invention may comprise a pharmaceutical composition comprising: therapeutic agent X or a pharmaceutically acceptable salt thereof; therapeutic agent Y or a pharmaceutically acceptable salt thereof; and at least one pharmaceutically acceptable An excipient where the therapeutic agent X is a non-steroidal anti-inflammatory drug and the therapeutic agent Y is a fatty acid oxidation inhibitor is acceptable. In a further embodiment, the present invention may include a method of treating a condition in a subject in need thereof, the method comprising administering to the subject an effective amount of a pharmaceutical composition having: therapeutic agent X or its pharmaceutically acceptable salt; therapeutic agent Y or a pharmaceutically acceptable salt thereof; and at least one pharmaceutically acceptable excipient, wherein the therapeutic agent X is a non-steroidal anti-inflammatory drug and the therapeutic agent Y is Fatty acid oxidation inhibitors.

In some embodiments, in the pharmaceutical composition, therapeutic agent X, or a pharmaceutically acceptable salt thereof, and therapeutic agent Y, or a pharmaceutically acceptable salt thereof, are contained in a single dosage form.

In a further embodiment, in the pharmaceutical composition, therapeutic agent X or a pharmaceutically acceptable salt thereof and therapeutic agent Y or a pharmaceutically acceptable salt thereof are present in separate dosage forms.

In some embodiments, the therapeutic agent X is selected from at least one of the following: COX inhibitors, salicylates, ibuprofen, phenoxyibuprofen, naproxen, diclofenac, celecoxib, Mefenamic acid, etoroxib, indomethacin, ketorolac, tetraclofenac, sulindac, and tolmetin. In some embodiments, therapeutic agent X is salicylate or a derivative thereof. In some embodiments, therapeutic agent X is 2-acetyloxybenzoic acid. In some embodiments, the therapeutic agent Y is selected from at least one of the following: CPT inhibitors, carnitine biosynthesis inhibitors, 3-ketoacyl-CoA thiolase inhibitors, etamoxir, hydroxy Phenylglycine, perhexiline, meldonium, 1-(2,3,4-trimethoxybenzyl)piperazine dihydrochloride, trimetazidine, ethoxycarnitine, aminocarnitine, or carnitine Phosphonooxy derivatives of bases. In some preferred embodiments, the therapeutic agent Y is 1-(2,3,4-trimethoxybenzyl)piperazine dihydrochloride.

In some embodiments, the mass ratio of therapeutic agent X to therapeutic agent Y is between 1:1 and 10:1, including 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, or 10:1. In some preferred embodiments, the mass ratio of therapeutic agents is 5:1 or 6:1 or 7:1.

In some embodiments, the amount of therapeutic agent X can range anywhere from 1 mg/kg to 100 mg/kg or more per day and the amount of therapeutic agent Y can range from 0.25 mg/kg to 100 mg/kg or more per day within any range. The exact amount will vary depending on the particular disease indication and the physiology of the individual. In some embodiments, therapeutic agent X and therapeutic agent Y may be administered once in n days, where n ranges from 1 to 14. In some embodiments, therapeutic agent X and therapeutic agent Y may be administered once a day or multiple times a day. In some embodiments, therapeutic agent X and therapeutic agent Y can be administered daily, every two days, every three days, every four days, every five days, every six days, every week, etc.

In some embodiments, therapeutic agent X and therapeutic agent Y are administered simultaneously. In some embodiments, therapeutic agent X and therapeutic agent Y are contained in a single dosage form. In some embodiments, possible routes of administration of therapeutic agent X and therapeutic agent Y in a single dosage form include, but are not limited to: oral, sublingual, buccal, nasal, inhalation, intratracheal, intravenous, intraarterial, coronary Intrathecal, intramuscular, intraperitoneal, intramyocardial, transendocardial, transepicardial, subcutaneous, transdermal, vaginal, rectal, or aural. In some preferred embodiments, therapeutic agent X and therapeutic agent Y are administered orally or intravenously or subcutaneously in a single dosage form.

In some embodiments, therapeutic agent X and therapeutic agent Y are administered separately. In some embodiments, therapeutic agent X is administered before therapeutic agent Y. In some embodiments, therapeutic agent X is administered after therapeutic agent Y. In some embodiments, possible routes of administration of Therapeutic Agent X and Therapeutic Agent Y in separate dosage forms include, but are not limited to: oral, sublingual, buccal, nasal, inhalation, intratracheal, intravenous, intraarterial, coronary Intrathecal, intramuscular, intraperitoneal, intramyocardial, transendocardial, transepicardial, subcutaneous, transdermal, vaginal, rectal, or aural. In some embodiments, therapeutic agent X and therapeutic agent Y are administered orally or intravenously in separate dosage forms.

In some embodiments, therapeutic agent X and therapeutic agent Y are administered orally separately. In some embodiments, therapeutic agent X and therapeutic agent Y are administered separately by injection.

Pharmaceutical compositions for parenteral injection include aqueous or nonaqueous solutions, dispersions, suspensions or emulsions, pharmaceutically acceptable sterile or non-sterile, and for use in sterile injectable solutions or dispersions Reconstituted powder.

In some embodiments, and for more efficient distribution, the compounds of the invention can be entrapped in prolonged controlled-release or targeted delivery systems, such as polymer matrices, liposomes, and microspheres.

In a preferred embodiment, the pharmaceutical composition of the present invention may be a solid composition for oral administration, specific examples of which include tablets, granules, fine granules, capsules, powders and pills. In addition to the therapeutic agent X or the pharmaceutically acceptable salt of the therapeutic agent Y or the combination thereof, the solid oral composition of the present invention can also add excipients, binders, lubricants, etc., and can be formulated into corresponding form. Examples of excipients that can be used include lactose, corn starch, crystalline cellulose, sucrose, glucose, mannitol, sorbitol and calcium carbonate. Examples of the binder include hydroxypropylcellulose, hydroxyethylcellulose, hypromellose, hydroxyethylethylcellulose, hydroxyethylmethylcellulose, polyvinylpyrrolidone, and polyvinylalcohol. Examples of lubricants include magnesium stearate, stearic acid, palmitic acid, calcium stearate and talc. Such formulations are described in detail in a number of sources well known and readily available to those skilled in the art. For example, Remington's The Science and Practice of Pharmacy, edited by Allen, Loyd V., Jr, 22nd ed., describes the preparation of formulations that can be used in conjunction with the present invention.

The following examples are intended purely as illustrations of the application and thus should not be considered limiting of the invention in any way. The following examples and detailed description are offered by way of illustration and not limitation.

Example 1: LPS-induced inflammation

Lipopolysaccharide (LPS), a common antigen expressed by many pathogenic microorganisms, has severe effects on the host response and cannot be treated with NSAIDs alone. In particular, LPS intraperitoneal treatment, a reliable model of pathogen-induced CRS, also recapitulates many features of human ARDS, including increased production of pro-inflammatory cytokines in the lung, high levels of immune infiltration, and changes in endothelial and vascular integrity. lost.

Adult DIO mice, 6-8 weeks old, were fed a high fat diet (Jackson Labs) for 12 weeks before starting any experiments. DIO mice received 5 mg/kg LPS (Sigma) intraperitoneally. Mice fed normal chow served as controls. Mice injected with PBS vehicle served as drug-treated controls. Mice were sacrificed 6 hours after stimulation with LPS (Fig. 1A). Lungs were frozen in liquid nitrogen for molecular assays or fixed in formalin for histological examination. Histological results showed severe pulmonary congestion, alveolar thickening, hyaline membrane formation, and inflammatory infiltration in the lung tissues of all LPS-treated mice, but these changes were more pronounced in DIO mice. Compared with normal mice, the lungs of DIO mice exhibited more severe signs of ALI, such as increased lung protein content, cell number in bronchoalveolar lavage fluid (BALF), and lactate dehydrogenase (LDH) in BALF content shown. These results indicated that diet-induced obese (DIO) mice were more susceptible to LPS-induced lung injury (Fig. 1B).

To test the effect on mortality and survival, LPS was injected intraperitoneally at a dose of 30 mg/kg. The mortality of mice was recorded every 12 hours for 3 days after LPS injection (n=12). LPS-treated DIO mice began to die within 16 hours, and about 80% of the mice died within 40 hours. In contrast, normal mice treated with LPS lived longer, with only about 40 percent dying within 40 hours. These results indicated that diet-induced obese (DIO) mice were more likely to die from LPS-induced inflammation (Fig. 1B).

Further assessment of inflammatory cell infiltration into the lungs revealed higher lung myeloperoxidase (MPO) activity and neutrophil numbers in the BALF in DIO mice compared with normal mice. TNF-α levels in BALF and lung tissue of DIO mice were also much higher than those of normal mice treated with LPS. These results suggest that obesity exacerbates LPS-induced pneumonia (Fig. 1C).

To test whether prophylactic treatment with drugs X and Y could block the pathogenesis of ALI, we performed a rescue experiment using intraperitoneal injections of X and Y. In this experiment, in the examples listed later, X is 2-acetyloxybenzoic acid (CAS No.: 50-78-2) and Y is 1-[(2,3,4-trimethoxybenzene yl)methyl]piperazine (CAS No.: 5011-34-7). Histological analysis revealed improved lung structure in DIO mice treated prophylactically with drug combination XY 2 weeks prior to LPS injection compared to DIO mice treated with X alone, Y alone, or PBS vehicle 2 weeks prior to LPS injection significantly improved. These results indicated that drug combination XY improved ALI in DIO mice (Fig. 1B).

Survival analysis further showed that DIO mice treated prophylactically with drug combination XY 2 weeks prior to LPS injection had improved survival compared to DIO mice treated with X alone, Y alone, or PBS vehicle 2 weeks prior to LPS injection. significantly improved. These results indicated that drug combination XY prevented ALI-induced death in DIO mice (Fig. 1B).

In addition, compared with DIO mice treated with X alone, Y alone, or PBS vehicle 2 weeks before LPS injection, edema forming proteins, cell numbers, LDH content and neutrophil count as well as MPO activity were significantly attenuated in BALF. These results indicated that drug combination XY ameliorated LPS-induced pneumonia (Fig. 1C).

Example 2: Viral co-receptors

To test whether drug treatment could effectively reduce the expression of viral co-receptors with potential efficacy in reducing viral infection, we performed RNAseq analysis of various organs after XY treatment for 3 days. Briefly, total RNA was purified by ethanol precipitation and sent for RNAseq. A paired-end library was constructed and sequenced using the Nova-PE150 platform (Illumina). Figures 2 and 3 show that drug combination XY reduces the expression of HCV co-receptors CD36 and CD81. Figures 4 and 5 show that the drug combination XY reduces the expression of the hepatitis A virus co-receptors HAVCR1 and HAVCR2. Figure 6 shows that the drug combination XY reduces the expression of the co-receptor ACE2 of SARS-CoV and SARS-CoV-2 and the co-receptor of MERS coronavirus DPP4. Figures 7-12 show that drug combination XY reduces the expression of Integrin α(v), Integrin β1, Integrin β3, Integrin β5, Integrin β6, which are co-receptors for adenovirus, parvovirus and cytomegalovirus. These results suggest that drug combination XY can attenuate the spread of viral infectious diseases.

Example 3: Senescent cells in aged mammalian tissues

Markers of mammalian cells exhibiting senescence are positive staining for senescence-associated-β-galactosidase (SA-β-gal), as well as protein expression of p16INK4A and senescence-associated heterochromatin markers such as H3K9me3. To test whether drugs X and Y can reduce tissue aging in aged mice, we injected 20 mg/kg of X, 4 mg/kg of Y, or 20 mg/kg intraperitoneally into 2-year-old aged mice (n=10/group). kg X and 4 mg/kg Y, relative to DMSO vehicle control, once daily for 30 days. Liver, muscle and skin tissue sections were stained for SA-β-gal activity using the Aging β-galactosidase Staining Kit (Cell Signaling Technology) according to the manufacturer's protocol. Protein expression of pl6INK4A and H3K9me3 was assessed by densitometric quantification of Western blots of liver, muscle and skin tissue samples. The results showed that only the drug combination X and Y significantly reduced all three biomarkers of aging in aged liver, muscle and skin tissues (P<0.01), but neither X nor Y alone ( 13A-13C). This suggests that drug combinations X and Y are able to reduce senescent cells and/or reverse aging.

Example 4: Skin Wound Healing and Tissue Repair During Aging

Mammals exhibit a decreased rate of tissue repair with aging. One animal model of this phenomenon is the delayed healing of skin wounds in aged db/db mice compared to young mice. Chronic wounds were generated in db/db mice by making full thickness 6 mm diameter excisional wounds on the dorsal skin of 9 month old mice. We used a well-described murine full-thickness excision wound model (Loh et al., 2009). Briefly, two circular full-thickness wounds were made on the dorsal skin of mice using a 6 mm disposable biopsy punch (Delasco). Silicone wound splints (Grace BioLabs) were closed with 4-0 nylon sutures to prevent skin contractures. Wounds were covered with sterile occlusive dressings and monitored daily. Monitor boundaries with frequent application of permanent markers. Photographs were taken at various time points throughout the experiment. Wounds remained open 20 days after trauma, sometimes more than 3 months, as expected for chronic wounds. To test whether drugs X and Y can treat chronic wounds in aged mice, we injected 20 mg/kg of X or 4 mg/kg of Y, or 20 mg of Both X at 4 mg/kg and Y at 4 mg/kg were administered once daily for 30 days relative to the DMSO vehicle control.

Quantification of the dorsal wound area showed that only the drug combination X and Y significantly accelerated tissue repair in aged mice with chronic wounds relative to the DMSO vehicle control (P<0.01), but neither X alone nor Y alone had this effect (Figure 14).

Example 5: Hair loss and hair regrowth during aging

A person exhibits hair loss or hair loss as they age, most notably in men exhibiting androgenetic alopecia or senile alopecia. Inflammation and aging, or inflammatory aging, are risk factors for hair loss. The animal model for this phenomenon is delayed hair regrowth in inflammatory-aged ob/ob mice following depilation. After removing back hair from 8-week-old mice using depilatory cream (day 0), 3 wild-type mice and 3 inflammatory-aged mice were monitored regularly until the depilated area was mostly covered with hair. The regrowth can be seen as dark hairs on pinkish-white shaved skin. Hair regrowth areas were quantified using the NIH ImageJ software program (NTIS, Springfield, VA). When the hair cycle of wild-type mice reaches the second telogen phase at 8 weeks of age, depilation induces a rapid transition to anagen at 1 week post-depilation and full hair growth at 4 weeks post-depilation in wild-type mice regeneration. In contrast, only the first signs of anagen were observed at 6 weeks after depilation in inflammatory-aged mice. To test whether drugs X and Y can ameliorate hair regeneration defects in inflammatory aging mice, we intraperitoneally injected 20 mg/kg of X or 4 mg/kg of Y in inflammatory aging mice (n=10/group) after depilation, Or both X at 20 mg/kg and Y at 4 mg/kg once daily for 30 days versus DMSO vehicle control. Quantification of dorsal skin areas with hair regrowth showed that only drug combination X and Y significantly accelerated hair regrowth after depilation in inflammatory aging mice relative to DMSO vehicle control (P<0.01), but not X alone or Y alone. Effect (Figure 15).

Example 6: Tissue Fibrosis and Tissue Degeneration During Aging

Aging mammals also exhibit a reduced ability to regress fibrosis, leading to tissue scarring and irreparable organ damage, such as in idiopathic pulmonary fibrosis (IPF), liver fibrosis, and cardiac fibrosis. The animal model of IPF is bleomycin-induced lung injury in mice. Young and aged mice were anesthetized with intraperitoneal injections of ketamine (100 mg/kg) and xylazine (10 mg/kg), followed by intratracheal administration of bleomycin (1.25 U/kg) to induce lung injury, or administration of saline (total volume 50 μL). Fibrosis was quantified by measuring fibrotic areas in lung tissue sections with Masson's trichrome staining. The animal model for liver fibrosis and cardiac fibrosis is the inflammatory aging mouse model, ob/ob mice fed a high-fat, high-glucose, high-cholesterol diet for 90 days (Research Labs). In order to test whether drugs X and Y can improve lung fibrosis in bleomycin-injured mice, and liver and heart fibrosis in inflammatory aging mice, we treated bleomycin-treated mice (n=10/group) and inflammatory aging mice (n=10/group) were injected intraperitoneally with 20 mg/kg of X, or 4 mg/kg of Y, or both 20 mg/kg of X and 4 mg/kg of Y, relative to the DMSO vehicle control, Once daily for 30 days. Quantification of fibrosis by Masson's trichrome staining showed that only drug combination X and Y significantly reversed fibrosis in inflammatory aged mice relative to DMSO vehicle control (P<0.01), but X alone or Y alone did not This effect (Figure 16).

Example 7: Inflammation and Death

Aging, especially instances of inflammatory aging, is also associated with an increased risk of death from septic shock or pro-inflammatory cytokine release syndrome. This is partly due to inflammatory aging-induced immune senescence, resulting in a dysfunctional immune response, which leads to multi-organ failure and death. The animal model for this phenomenon is LPS endotoxin-induced septic shock in aged mice. For lethality studies, LPS (Sigma-Aldrich) was injected intraperitoneally at a dose of 30 mg/kg. Mortality of mice was recorded every 12 hours for 3 days after LPS challenge in each group. To test whether drugs X and Y could prophylactically ameliorate the lethal effect of LPS-induced sepsis, we injected 20 mg/kg of X or 4 mg/kg of Y intraperitoneally into 2-year-old mice (n=10/group), or Both X at 20 mg/kg and Y at 4 mg/kg were given once daily for 30 days relative to the DMSO vehicle control before they were challenged with LPS. Quantification of survival over time showed that only the drug combination X and Y significantly improved survival after LPS challenge relative to the DMSO vehicle control (P<0.01), but neither X nor Y alone had this effect ( Figure 17).

Example 8: Gene signatures related to immune aging

Aging, especially chronic inflammatory aging, leads to immunosenescence, the reduction of naive T cells and the dysfunction of immune cell subsets with aging. When immune challenged, this results in an abnormal surge of Th17- and Th1-driven pro-inflammatory responses at the expense of Treg- and Th2-driven anti-inflammatory responses, which can lead to fatal cytokine release syndrome and/or sepsis. To test how the drug combination of X and Y affects the immune response, we isolated peripheral blood mononuclear cells from mice treated daily for 30 days with 20 mg/kg of X and 4 mg/kg of Y versus DMSO vehicle control (PBMC), and their RNA was extracted with Trizol (Invitrogen) for RNAseq (Illumina). Following Gene Set Enrichment Analysis (GSEA; Broad Institute), our results show that drug combination X and Y inhibit Th1 pro-inflammatory signature, Th1-associated/pro-Th17 cytokine IL2 signature, Th1/Th17-associated cells Factor IFNG signature, and pan-activated Th-associated cytokine IL3/IL5/GM-CSF signature (Figures 18A and 18B). In contrast, X and Y promoted the Th2/Treg-associated anti-inflammatory cytokine profile of IL4, IL10 and IL13 (Fig. 18C).

Example 9: Mimicking Exercise to Inhibit Inflammatory Aging

Physical muscle building is known to ameliorate the effects of aging, including chronic inflammatory aging and immune aging. To test how drug combinations X and Y affect the effects of physical exercise, type I slow-twitch muscle fibers were immunostained on frozen sections of the quadriceps muscles of inflamed aged mice (Fig. 19A) and examined by fluorescence microscopy (Carl Zeiss )). The results showed that drug combination X and Y more than doubled the type I muscle fiber area (P<0.01), indicating that drug combination X and Y mimicked and enhanced the effect of endurance exercise ( FIG. 19B ). Furthermore, quantification of muscle fiber cross-sectional area revealed that drug combination X and Y increased the proportion of muscle fibers whose area exceeded the 100-pixel threshold (P<0.01), suggesting that drug combination X and Y induced muscle hypertrophy like physical exercise (Fig. 19C). Finally, grip strength was measured as the average of 6 maximum peak strength measurements generated on a grip dynamometer (Bioseb) and normalized to their body weight. The results showed that drug combination X and Y significantly increased the grip strength of inflammatory aging mice (P<0.01), indicating that drug combination X and Y increased limb muscle strength like physical exercise ( FIG. 19D ).

Example 10: Mimicking Exercise Induces Secreted Factors to Suppress Inflammatory Aging

Physical exercise is known to ameliorate the effects of aging, including chronic inflammatory aging and immunosenescence, by promoting the secretion of pro-metabolic and immunomodulatory cytokines. To test how drug combination X and Y affect the effect of physical exercise through such secreted factors, the muscle of inflammatory aging mice was analyzed using RNAseq, and the serum of inflammatory aging mice was analyzed using ELISA (R&D Corporation). The results showed that drug combination X and Y significantly increased multiple secreted factors, including myokines (Figure 20), including Angptl8, Angptl3, Angptl4, adiponectin, BDNF, IGF1, IL6, irisin and LIF (P<0.01), Drug combinations X and Y are shown to affect aging and inflammatory aging at least in part by mimicking the effects of physical exercise and inducing a number of secreted factors including myokines.

Example 11: Mimicking calorie restriction-induced anti-aging gene pathways

Calorie restriction is known to ameliorate the effects of aging by altering cellular bioenergetic stress and promoting increased mitochondrial activity, unfolded protein response (UPR), secretion of beneficial cytokines, and stem cell regeneration. To test how drug combination X and Y mimic the effects of calorie restriction (CR), we analyzed the metabolism of primary human skeletal muscle cells treated with X (100 mg/L) and Y (2 mg/L) at passage 15, compared with X alone (100mg/L) or Y alone (2mg/L). The results showed that X and Y directly interacted with the mitochondrial proteins UQCRH, NDUFS6 and COX7A2 according to thermal proteomic analysis using the TMTpro 16plex kit and Q-Exactive HF-X mass spectrometry (Thermo Fisher; Figures 21A-21C ). Combined, synergistically reduces ATP levels within minutes (Fig. 22), synergistically reduces acetyl-CoA and protein acetylation (Fig. 23), and synergistically increases Parkin-driven mitophagy according to Ab23707 antibody immunostaining of Pink1 ( Figure 24), synergistically increased mitochondrial targeting protein levels (Figure 25A and 25B), according to qRT-PCR, relative to the nuclear genome B2M synergistically increased mitochondrial DNA levels for mitochondrial genomes ND1 and ND4 (Figure 26), according to RNAseq (100 million Minda HiSeq) and gene set enrichment analysis (GSEA) synergistically increased IRE1a-mediated unfolded protein response (UPR) (Figure 27) and synergistically decreased mitochondrial reactive oxygen species (ROS; Figures 28A and 28B). According to gene set enrichment analysis (GSEA) (Fig. 29A-29C), X and Y synergistically increase the activity of multiple cytokine pathways, including NGF, GDNF and axon guidance factor, suggesting that the drug combination X and Y stimulate the neuroendocrine system And simulate calorie restriction to delay aging. Finally, drug combination X and Y synergistically increased the expression of stem cell pathways (e.g., Hippo, Notch-Hes-Hey, HIF1a, and Integrin-MAPK) according to RNAseq (Hiseq) and gene set enrichment analysis (GSEA) (Fig. 30A–30E), indicating increased stemness and regenerative capacity. These beneficial anti-aging effects were not observed with either X alone or Y alone or vehicle.

Example 12: Induction of anti-aging, pro-regenerative responses in vivo

Application of drug combination X and Y in cynomolgus monkeys increases neuroendocrine cytokines like calorie restriction, thereby increasing antiaging-associated muscle regeneration and reducing aging-associated central obesity. By dissolving X and Y in DURO-TAK87-2677PSA (Pressure Sensitive Adhesive) solution, and coating it on the release film (Scotchpak ^TM 1022), then baking and drying it with the back film (Scotchpak ^TM 1109) Laminated, we constructed pressure-sensitive adhesive dispersed patches for controlled transdermal release of drug X and Y every 7 days. After 2 rounds of transdermal drug treatment, we found that total body weight and waist circumference (indicators of aging-induced central obesity) in cynomolgus monkeys both decreased significantly by about 10-20%, while calf circumference (indicators of skeletal muscle growth and regeneration) An increase of about 8% (Fig. 31). Importantly, plasma levels of their neuroendocrine cytokines BDNF and LIF (R&D Systems ELISA) were also significantly increased after treatment with the drug combination X and Y, but not X alone or Y alone or vehicle ( 32A and 32B). No toxicity was observed based on clinical hematology or clinical blood chemistry (Figures 33A and 33B).

Drug combination X (20 mg/kg) and Y (4 mg/kg) relative to X alone (20 mg/kg) or Y alone (4 mg/kg) or vehicle (phosphate-buffered saline or DMSO in PBS) Daily oral gavage over 6 weeks leads to increased antiaging-associated tissue regeneration in 1-year-old diabetic mice. In their livers with age-associated steatohepatitis, X and Y promoted liver regeneration as observed by increased hepatocyte proliferation indicated by Ki67 (Ab16667) immunostaining (Figure 34). In their pancreas with age-related pancreatitis due to type 2 diabetes, X and Y reduced beta islet fibrosis according to Masson's trichrome staining (Fig. 35) and immunostaining according to Ki67 (Ab16667) (Fig. 36A and 36B) and qRT-PCR targeting Pax6, Mafa, Pdx1 and Ins using primers from Origen (Figure 37A and 37B) increased beta islet cell growth and proliferation, thereby promoting pancreatic beta islet regeneration. In their skin, X and Y promoted senescent hair regrowth and wound healing at 64 days after shaving and wounding with a dorsal skin punch (10 mm diameter) (Figure 38). In their quadriceps muscles, X and Y increased myosin heavy chain (MHC; Sigma-Aldrich - M4276 antibody) protein levels (Fig. 39), thereby promoting skeletal muscle regeneration. Furthermore, total body weight was significantly increased compared to aged control animals with muscle atrophy and sarcopenia, indicating that X and Y reverse sarcopenia (Figure 40). These beneficial anti-aging effects were not observed with either X alone or Y alone or the vehicle control.

Daily oral gavage of the drug combination X and Y over 3 weeks resulted in significantly higher levels of intratracheal bleomycin (3 mg/kg in 50 ul phosphate-buffered saline or PBS) compared to X alone or Y alone or vehicle. ) in 1-year-old mice treated to mimic chronic obstructive pulmonary disease (COPD) and idiopathic pulmonary fibrosis (IPF), decreased age-related pulmonary fibrosis. Masson's trichrome staining and ImageJ (NIH) quantification of the percentage of fibrotic area indicated that the level of lung fibrosis was significantly reduced in mice treated with X and Y (Figure 41). These beneficial anti-aging effects were not observed with either X alone or Y alone or the vehicle control. Daily oral gavage of drug combination X and Y over 6 weeks resulted in treatment with subcutaneous dehydroepiandrosterone (DHEA; 60 mg/kg body weight) and ad libitum feeding of high fat- Restoration of the estrous cycle in adult female rats fed a high-sugar-high-cholesterol diet to mimic premature ovarian failure in polycystic ovary syndrome (PCOS). Vaginal smear analysis showed that most of the female rats treated with X and Y returned to the estrous cycle (Figure 42). These beneficial anti-aging effects were not observed with X alone or Y alone or vehicle controls in which all rats were frozen in estrus or proestrus.

While the invention has been illustrated and described herein with reference to preferred embodiments and specific examples thereof, it will be apparent to those of ordinary skill in the art that other embodiments and examples can perform a similar function and/or achieve a similar result. All such equivalent embodiments and examples are within the spirit and scope of the invention, are thereby contemplated and intended to be covered by the appended claims.

Claims (30)

1. A composition of matter for use in preventing or treating a microbial infection, the composition of matter comprising a combination of a therapeutic agent X and acceptable salts thereof and a therapeutic agent Y and acceptable salts thereof, wherein the therapeutic agent X is a non-steroidal anti-inflammatory drug and the therapeutic agent Y is a fatty acid oxidation inhibitor.

2. A composition of matter for use in the prevention or treatment of an inflammatory disorder associated with a microbial infection, the composition of matter comprising a combination of a therapeutic agent X and acceptable salts thereof and a therapeutic agent Y and acceptable salts thereof, wherein the therapeutic agent X is a non-steroidal anti-inflammatory drug and the therapeutic agent Y is a fatty acid oxidation inhibitor.

3. The composition of matter of any one of claims 1 and 2, for use in the prevention or treatment of microbial infections and related inflammatory disorders by modulating host responses.

4. A composition of matter as claimed in any one of claims 1 to 3 for use in the prevention or treatment of microbial infections and related inflammatory disorders by modulating the host immune system.

5. A composition of matter as claimed in any one of claims 1 to 3 for use in the prevention or treatment of microbial infections and related inflammatory disorders by modulating host inflammation.

6. A composition of matter as claimed in any one of claims 1 to 3 for use in the prevention or treatment of microbial infections and related inflammatory disorders by reduction of viral receptors.

7. The composition of matter of any one of claims 1 to 6, for use in the prevention or treatment of an inflammatory disorder selected from the group consisting of: CRS, ARDS, ALI, AIDS, asthma, chronic peptic ulcer, tuberculosis, rheumatoid arthritis, periodontitis, ulcerative colitis, crohn's disease, sinusitis, encephalitis, myelitis, meningitis, arachnoiditis, neuritis, dacryocystitis, scleritis, episcleritis, keratitis, retinitis, chorioretinitis, blepharitis, conjunctivitis, uveitis, otitis externa, otitis media, labyrinthine, mastoiditis, cardiac inflammation, endocarditis, myocarditis, pericarditis, vasculitis, arteritis, phlebitis, capillary vasculitis, sinusitis, rhinitis, pharyngitis, laryngitis, tracheitis, bronchitis, bronchiolitis, pneumonia, pleurisy, mediastinitis, stomatitis, gingivitis, glossitis, tonsillitis, salivary gland inflammation, parotitis, cheilitis, pulpitis, jaw inflammation, esophagitis, gastritis, gastroenteritis, enteritis, colitis, enterocolitis duodenitis, ileitis, cecitis, appendicitis, proctitis, hepatitis, ascending cholangitis, cholecystitis, pancreatitis, peritonitis, dermatitis, folliculitis, cellulitis, sweat gland inflammation, arthritis, dermatomyositis, myositis, synovitis, tenosynovitis, bursitis, attachment point inflammation, fasciitis, cystitis, epicondylitis, tendinitis, panniculitis, osteochondritis, osteomyelitis, spondylitis, tendinitis, inflammation of the liver, and the like periostitis, chondritis, nephritis, glomerulonephritis, pyelonephritis, ureteritis, cystitis, urethritis, oophoritis, salpingitis, endometritis, parahysteritis, cervicitis, colpitis, vulvitis, mastitis, orchitis, epididymitis, prostatitis, seminal vesicle, balanitis, dermatitis of the package, balanitis, chorioamniosis, umbilical cord inflammation, islet inflammation, pituitary inflammation, and the like, thyroiditis, parathyroid, adrenalitis, lymphangitis or lymphadenitis.

8. The composition of matter of any one of claims 1 to 7 for use in the prevention or treatment of a microbial infection caused by a bacterium, virus, fungus, parasite or protozoan.

9. The composition of matter of any one of claims 1 to 8 for use in the prevention or treatment of a microbial infection caused by a virus.

10. The composition of matter of any one of claims 1 to 9, for use in the prevention or treatment of a microbial infection caused by a member of the group consisting of: adenoviridae, papovaviridae, parvoviridae, dactyloviridae, polymorphous enveloped viridae, myoviridae, brachytheraviridae, longueviridae, isopsorviridae, herpesviridae (including human herpesviruses, and varicella zoster viruses), maroviridae, lipophagosphaidae, archaeolidae, adenoviridae, bottle viridae, vesicular viridae, african swine fever viridae (including african swine fever viruses), baculoviruses, chikataviridae, rhabdoviridae, cover phage, spiny phage, globular viridae, trichoviridae, mastadeviridae, iridoviridae, mosaic viridae, pseudoviridae, nude viridae, linear viridae, panaviridae, papillomaviridae algae DNA virus family, budding phage family, polyndna virus, polyomavirus family (including simian virus 40, JC virus, BK virus), poxvirus family (including vaccinia and smallpox virus), globoviridae, multi-layer virus family, californicae, dinonodna virus, salix end protein virus genus, or rez virus, coronaviridae, picornaviridae, reoviridae, calicivviridae, flaviviridae, togaviridae, vitroneviridae, filoviridae, paramyxoviridae, pneumoviridae, rhabdoviridae, arenaviridae, bunyaviridae, astroviridae, orthomyxoviridae, arteriviridae, hepaciviridae, retrovirus, caulifloviridae, hepadnaviridae or delta virus, or combinations thereof.

11. The composition of matter of any one of claims 1 to 10, wherein the viral infection is caused by: coronavirus, cytomegalovirus, poliovirus, rhinovirus, hepatitis A, hepatitis B, norwalk virus, yellow fever virus, west Nile virus, hepatitis C virus, dengue virus, zika virus, rubella virus, ross river virus, sindbis virus, chikungunya virus, boerna disease virus, ebola virus, marburg virus, measles virus, mumps virus, nipa virus, hendela virus, newcastle disease virus, human respiratory syncytial virus, human immunodeficiency virus, rabies virus, lassa virus, hantaan virus, critiferam-Congo hemorrhagic fever virus, influenza, or hepatitis D virus.

12. The composition of matter of any one of claims 1 to 11, wherein therapeutic agent X or an acceptable salt thereof and therapeutic agent Y or an acceptable salt thereof are contained in a single dosage form.

13. The composition of matter of any one of claims 1 to 12, wherein therapeutic agent X or an acceptable salt thereof and therapeutic agent Y or an acceptable salt thereof are contained in separate dosage forms.

14. The composition of matter of any one of claims 1 to 13, wherein the mass ratio of therapeutic agent X to therapeutic agent Y is 1:1 to 10:1, including 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, or 10:1.

15. The composition of matter of any one of claims 1 to 14, wherein the route of administration to a subject comprises oral, sublingual, buccal, nasal, inhalation, intratracheal, intravenous, intraarterial, intracoronary, intrathecal, intramuscular, intraperitoneal, intramyocardial, endocardial, epicardial, subcutaneous, transdermal, vaginal, rectal or otic.

16. The composition of matter of any one of claims 1 to 15, wherein therapeutic agent X is selected from at least one of the following: COX inhibitors, salicylates, ibuprofen, phenoxyibuprofen, naproxen, diclofenac, celecoxib, mefenamic acid, etoricoxib, indomethacin, ketorolac, tetrachlorofenac, sulindac, and tolmetin.

17. The composition of matter of any one of claims 1 to 15, wherein therapeutic agent Y is selected from at least one of the following: CPT inhibitors, carnitine biosynthesis inhibitors, 3-ketoacyl CoA thiolase inhibitors, etorick house, hydroxyphenylglycine, perhexiline, mildrozide, trimetazidine, ethoxycarnitine, aminocarnitine or phosphonooxy derivatives of carnitine.

18. A composition of matter for preventing or treating aging or an aging-related disorder, the composition of matter comprising a combination of a therapeutic agent X and an acceptable salt thereof and a therapeutic agent Y and an acceptable salt thereof, wherein the therapeutic agent X is a non-steroidal anti-inflammatory drug and the therapeutic agent Y is a fatty acid oxidation inhibitor.

19. A composition of matter for use in aging or aging-related disorders by promoting regeneration, the composition of matter comprising a combination of a therapeutic agent X and acceptable salts thereof and a therapeutic agent Y and acceptable salts thereof, wherein the therapeutic agent X is a non-steroidal anti-inflammatory drug and the therapeutic agent Y is a fatty acid oxidation inhibitor.

20. A composition of matter for aging or aging-related disorders by reversing fibrosis, the composition of matter comprising a combination of a therapeutic agent X and acceptable salts thereof and a therapeutic agent Y and acceptable salts thereof, wherein the therapeutic agent X is a non-steroidal anti-inflammatory drug and the therapeutic agent Y is a fatty acid oxidation inhibitor.

21. A composition of matter for use in aging or aging-related disorders by reversing aging, the composition of matter comprising a combination of a therapeutic agent X and an acceptable salt thereof and a therapeutic agent Y and an acceptable salt thereof, wherein the therapeutic agent X is a non-steroidal anti-inflammatory drug and the therapeutic agent Y is a fatty acid oxidation inhibitor.

22. A composition of matter for use in aging or aging-related disorders by improving insulin sensitivity, the composition of matter comprising a combination of a therapeutic agent X and an acceptable salt thereof and a therapeutic agent Y and an acceptable salt thereof, wherein the therapeutic agent X is a non-steroidal anti-inflammatory drug and the therapeutic agent Y is a fatty acid oxidation inhibitor.

23. The composition of matter of claims 18 to 22 for preventing or treating aging or aging-related disorders by modulating inflammation.

24. The composition of matter of claims 18 to 23, which prevents or treats aging and aging-related disorders selected from the group consisting of: alzheimer's disease, parkinson's disease, dementia atherosclerosis, cardiovascular diseases, type II diabetes, cancer, arthritis, rheumatoid arthritis, periodontitis, cataracts, osteoporosis, fibrosis, cirrhosis, idiopathic pulmonary fibrosis, scarring, joint fibrosis, chronic kidney disease, crohn's disease, keloids, myelofibrosis, retroperitoneal fibrosis, scleroderma, cirrhosis, chronic wounds, diabetic foot ulcers, skin aging, non-alcoholic steatohepatitis (NASH), menopause, hair loss, and sarcopenia.

25. The composition of matter of any one of claims 18 to 28, wherein therapeutic agent X or an acceptable salt thereof and therapeutic agent Y or an acceptable salt thereof are contained in a single dosage form.

26. The composition of matter of any one of claims 18 to 29, wherein therapeutic agent X or an acceptable salt thereof and therapeutic agent Y or an acceptable salt thereof are contained in separate dosage forms.

27. The composition of matter of any one of claims 18 to 30, wherein the mass ratio of therapeutic agent X to therapeutic agent Y is 1:1 to 10:1, including 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, or 10:1.

28. The composition of matter of any one of claims 18 to 30, wherein the route of administration to the subject comprises oral, sublingual, buccal, nasal, inhalation, intratracheal, intravenous, intraarterial, intracoronary, intrathecal, intramuscular, intraperitoneal, intramyocardial, endocardial, epicardial, subcutaneous, transdermal, vaginal, rectal or otic.

29. The composition of matter of any one of claims 18 to 28, wherein therapeutic agent X is selected from at least one of: COX inhibitors, salicylates, ibuprofen, phenoxyibuprofen, naproxen, diclofenac, celecoxib, mefenamic acid, etoricoxib, indomethacin, ketorolac, tetrachlorofenac, sulindac, and tolmetin.

30. The composition of matter of any one of claims 18 to 29, wherein therapeutic agent Y is selected from at least one of the following: CPT inhibitors, carnitine biosynthesis inhibitors, 3-ketoacyl CoA thiolase inhibitors, etorick house, hydroxyphenylglycine, perhexiline, mildrozide, trimetazidine, ethoxycarnitine, aminocarnitine or phosphonooxy derivatives of carnitine.