CN117298247A

CN117298247A - Use of MDP analogues in the manufacture of a medicament for the treatment of inflammatory bowel disease

Info

Publication number: CN117298247A
Application number: CN202210708930.7A
Authority: CN
Inventors: 周宏伟; 何肖龙; 黄祯荷; 胡梦瑶; 高杰
Original assignee: Southern Medical University Zhujiang Hospital
Current assignee: Southern Medical University Zhujiang Hospital
Priority date: 2022-06-22
Filing date: 2022-06-22
Publication date: 2023-12-29

Abstract

The present application relates to the use of MDP analogues in inflammatory bowel disease. The application discloses MDP analogues for the first time, which have effective anti-inflammatory effect and inhibition effect on inflammatory cytokine production, can effectively treat and prevent inflammatory bowel diseases, and provide a new drug treatment scheme for clinical treatment of IBD.

Description

Use of MDP analogues in the manufacture of a medicament for the treatment of inflammatory bowel disease

Technical Field

The application relates to the field of biological medicine, in particular to application of MDP analogues in preparation of medicines for treating inflammatory bowel diseases.

Background

Inflammatory bowel disease (inflammatory bowel disease, IBD) is an autoimmune bowel disease characterized by chronic inflammatory reactions and gastrointestinal epithelial lesions, the pathogenesis of which is not yet defined, and is characterized by relief of intestinal inflammation alternating with recurrence, mainly including ulcerative colitis (Ulcerative Colitis, UC) and Crohn's Disease (CD). UC is a chronic nonspecific colon inflammation, and lesions mainly involve mucous membranes of the colon, and the range of the lesions is from the far-end colon, so that the lesions can develop proximally and even involve the whole colon; the clinical manifestations are diarrhea, abdominal pain and mucopurulent bloody stool. CD is a chronic granulomatous inflammation, lesions may involve various parts of the gastrointestinal tract but are well developed in the terminal ileum and right half colon; the clinical manifestations are abdominal pain, diarrhea, intestinal obstruction (Peixoto Armando, ACG Case Reports Journal,2017,4 (1): e 46).

At present, no medicine capable of completely curing IBD exists, and the main purpose of the medicine for treating is to relieve symptoms and improve the life quality of patients clinically. Therapeutic agents include aminosalicylic acid formulations, glucocorticoids, immunosuppressants and various biological agents (e.g., TNF-alpha mab, etc.). The above drugs have disadvantages such as the susceptibility of aminosalicylic acid drugs to drug resistance in patients, recurrence of glucocorticoid after withdrawal, high cost of TNF antibody production and treatment, susceptibility to failure after prolonged use, and susceptibility to infection in patients (Paolo Gionchetti, dig Liver Dis,2017,49 (6): 604-17). Therefore, finding a safer and more effective anti-colitis drug is one of the hot spots of current IBD research, and is currently not meeting the urgent clinical demands.

Disclosure of Invention

The mechanism of IBD production is not known, the intracellular recognition receptor NOD2 is a key receptor in the development process of IBD, and its main ligand is bacterial cell wall peptidoglycan conserved fragment Muramyl Dipeptide (MDP), and after MDP is combined with NOD2, the signal pathways NF- κb and MAPK are activated, so that inflammatory mediator release is promoted, immune response is regulated, and colitis is relieved. MDP is a water-soluble small molecule, however, and has poor cell permeability and is easily cleared by the body, so that a more suitable therapeutic method for drugs needs to be found.

The inventors of the present application have found for the first time that Mifamurtide unexpectedly can show an effective effect of treating and preventing inflammatory bowel diseases, has an effective anti-inflammatory effect and an inhibitory effect on inflammatory cytokines (such as IL-6) and provides a new drug treatment scheme for clinical treatment of IBD.

In one aspect, the application provides an application of Mifamurtide in preparing a medicament for treating and/or preventing inflammatory bowel disease.

In certain embodiments, wherein the inflammatory bowel disease comprises ulcerative colitis and crohn's disease.

In certain embodiments, wherein the inflammatory bowel disease comprises sodium dextran sulfate (DSS) -induced Inflammatory Bowel Disease (IBD).

In another aspect, the present application provides the use of Mifamurtide in the manufacture of a medicament for inhibiting secretion of an inflammatory factor in the gut.

In certain embodiments, wherein the intestinal inflammatory factor is to include IL-1β, IL-1α, TNF α, GM-CSF, IL-6, and/or IL-17.

In certain embodiments, the primary active ingredient of the medicament is Mifamurtide.

In certain embodiments, the only active ingredient of the medicament is Mifamurtide.

In certain embodiments, wherein the medicament is formulated for administration by oral, inhalation, nasal spray, injection, topical, eye drop, rectal and/or vaginal administration.

In certain embodiments, wherein the medicament is formulated as a solid dosage form, a solution dosage form, a nebulizer aerosol dosage form, an injection dosage form, an ointment dosage form, a skin patch dosage form, a film dosage form, a soft capsule dosage form, or a suppository dosage form.

In certain embodiments, wherein the medicament is formulated for administration by oral, inhalation, nasal spray, injection, topical, eye drop, rectal and/or vaginal administration and in solid dosage forms, solution dosage forms, nebulizer aerosol dosage forms, injection dosage forms, ointment dosage forms, skin patch dosage forms, film dosage forms, soft capsule dosage forms, suppository dosage forms.

In certain embodiments, the medicament is formulated as tablets, capsules, granules, suppositories, ointments, patches, water injection and sustained and controlled release agents by oral, parenteral route or implantation.

In certain embodiments, the medicament is formulated as a pulmonary or nasal inhalation aerosol, metered dose aerosol or dry powder inhaler.

In certain embodiments, the medicament is formulated as a liquid injection or infusion.

In another aspect, the present application provides a medicament for treating inflammatory bowel disease comprising Mifamurtide or a pharmaceutically acceptable salt, ester, hydrate thereof, and optionally a pharmaceutically acceptable carrier.

In certain embodiments, wherein the medicament inhibits secretion of an inflammatory factor in the gut.

In certain embodiments, the effective dose of Mifamurtide or a pharmaceutically acceptable salt, ester, hydrate thereof is 0.01mg/kg to 100mg/kg.

Other aspects and advantages of the present application will become readily apparent to those skilled in the art from the following detailed description. Only exemplary embodiments of the present application are shown and described in the following detailed description. As those skilled in the art will recognize, the present disclosure enables one skilled in the art to make modifications to the disclosed embodiments without departing from the spirit and scope of the invention as described herein. Accordingly, the drawings and descriptions herein are to be regarded as illustrative in nature and not as restrictive.

Drawings

The specific features of the invention related to this application are set forth in the appended claims. The features and advantages of the invention that are related to the present application will be better understood by reference to the exemplary embodiments and the drawings that are described in detail below. The brief description of the drawings is as follows:

FIGS. 1A-1D show that different concentrations of Mifamurtide can improve DSS-induced inflammatory bowel disease. (a) Mifamurtide relieves weight loss; (B) Mifamurtide improves colon length reduction; (C) Mifamurtide reduces colon histopathological scores; (D) Mifamurtide reduces inflammatory factor IL-6 levels. Wherein P is equal to or less than 0.05, P is equal to or less than 0.01, and P is equal to or less than 0.001.

Mifamurtide, shown in FIGS. 2A-2C, for different routes of administration, improved DSS-induced inflammatory bowel disease. (2A) Mifamurtide relieves weight loss; (2B) Mifamurtide improves colon length reduction; (2C) Mifamurtide reduces colon histopathological scores; wherein P is equal to or less than 0.05, P is equal to or less than 0.01, and P is equal to or less than 0.001.

Fig. 3A to 3B show the comparison of the therapeutic effects of intraperitoneal injection of Mifamurtide and MDP: (3A) weight loss; (3B) serum FITC-dextran; wherein P is equal to or less than 0.05, P is equal to or less than 0.01, and P is equal to or less than 0.001.

Fig. 4A to 4C show HE staining results of different organs of mice after administration of high concentrations of Mifamurtide: (4A) liver; (4B) kidneys; (4C) small intestine.

Detailed Description

Further advantages and effects of the invention of the present application will become apparent to those skilled in the art from the disclosure of the present application, from the following description of specific embodiments.

Definition of terms

In the present application, the term "inflammatory bowel disease" (inflammatory bowel disease; IBD) generally refers to a refractory disease in which chronic inflammation and/or ulceration occurs in the mucosa of the intestinal tract, and as a representative example, the inflammatory bowel disease may be ulcerative colitis (Ulcerative colitis), crohn's disease, collagenous colitis, lymphocolitis, ischemic colitis, convertible colitis, behcet's syndrome, non-defined colitis, hemorrhagic rectal ulceration or ileal cystitis, but is not limited thereto.

In the present application, the term "ulcerative colitis" generally refers to a disease that continuously forms ulcers (erosion) or persistent inflammation of the mucosa of the large intestine, ulcers affecting the mucosal epithelium and the mucosa lamina propria (lamina) of the colon and rectum, 95% of ulcerative colitis affecting the rectum, and continuously spreads in the circumferential direction in the portion closer to the large intestine. Symptoms of ulcerative colitis include abdominal pain, bloody stool, mucobloody stool, diarrhea, and in severe cases, systemic symptoms such as fever, weight loss, anemia, etc.

In the present application, the term "crohn's disease" generally refers to a disease in which lesions such as ulcers are discontinuously formed in any part of the digestive tract from the oral cavity to the anus, and in addition to abdominal pain, diarrhea, bloody stool, in severe cases, symptoms such as fever, weight loss, general fatigue, anemia, and the like are also present.

In the present application, the term "intestinal inflammatory factor" generally refers to a pro-inflammatory cytokine that induces and/or promotes an inflammatory response in the intestinal tract. In some embodiments, the intestinal inflammatory factor comprises IL-1β, IL-1α, TNF α, IL-6, IL 17, GM-CSF, or any combination thereof.

In the present application, the term "muramyl dipeptide" (MDP, N-acetyl muramyl-L-alanyl-D-isoglutamine) generally refers to a synthetic immunoreactive peptide consisting of an N-acetyl muramic acid attached to a short amino acid chain of L-Ala-D-isogin. It was first identified in bacterial cell wall peptidoglycan (bacterial cell wall peptidoglycan) as the active component in Freund's Complete Adjuvant (FCA). In 1974, MDP was found to be one of the most effective and most widely used adjuvants in animal experimental models, the smallest structure required for efficacy of FCA. Muramyl dipeptide are well known for their immunomodulatory properties. Muramyl dipeptide is the smallest bioactive component of the bacterial cell wall. Muramyl dipeptide derivatives have been shown to exhibit significant immunomodulatory properties via one of PRR (pattern recognition receptor), nucleotide binding oligomerization domain 2 (N0D 2) receptors (Girardin S. Et al, 2003.J Biol Chem.27811.8869_72;F.Coulombe et al, 2012P1 et S0 ne,7 (5): arc ID e 36734). Muramyl dipeptide activates macrophages and other cells of the immune system to kill cancer cells (I.Jakopin, 2013.Current Medicinal Chemistry,20 (16): 2068-2079; ogawa et al, 2011.CurrBioact Compd.7 (3): 180-197), however, muramyl dipeptide has also been reported to be pyrogenic in nature. In order to reduce its pyrogenic effect, a series of MDP derivatives have been designed, synthesized and tested so far with the aim of enhancing specific functions while inhibiting pyrogenicity. There are many reported inventions or publications related to the synthesis of muramyl dipeptide and its derivatives (Namba et al, 1997. Vaccine,15,:405-13;W01996001645;US 4395399;US 7173107 B2)

In the present application, the term "analogue" generally refers to a second organic or inorganic molecule having a similar or identical function to, and being structurally similar to, the first organic or inorganic molecule in the context of a non-protein analogue. In some embodiments, the analog of Muramyl Dipeptide (MDP) may be the fatty acid derivative of MDP, MTP-PE. In other embodiments, the analog of MDP may be the amino acid derivative threonyl-MDP of MDP.

In this application, the term "pharmaceutically acceptable" generally refers to one or more non-toxic substances that do not interfere with the effectiveness of the biological activity of the active ingredient. Such formulations may generally contain salts, buffers, preservatives, compatible carriers and optionally other therapeutic agents. Such pharmaceutically acceptable formulations may also generally comprise compatible solid or liquid fillers, diluents or encapsulating materials suitable for administration to humans. For pharmaceutical use, the salt should be a pharmaceutically acceptable salt, but non-pharmaceutically acceptable salts may be conveniently used to prepare pharmaceutically acceptable salts, and they are not excluded from the scope of the present invention. Such pharmacologically and pharmaceutically acceptable salts include, but are not limited to, salts prepared from the following acids: hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, maleic acid, acetic acid, salicylic acid, citric acid, boric acid, formic acid, malonic acid, succinic acid, and the like. Pharmaceutically acceptable salts may also be prepared as alkali metal salts or alkaline earth metal salts, such as sodium, potassium or calcium salts.

In this application, the term "pharmaceutically acceptable carrier" generally refers to any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like, that are compatible with pharmaceutical administration. The use of such media and agents for pharmaceutically active substances is well known in the art. Any conventional medium or agent, unless incompatible with the active compound, is contemplated for use in the compositions of the present invention. Pharmaceutically acceptable carriers can also be included as pharmaceutically acceptable salts, wherein the term "pharmaceutically acceptable salts" includes salts of the active compounds prepared with relatively non-toxic acids or bases, depending on the particular substituents found on the compounds described herein. When the compounds of the present invention contain relatively acidic functional groups, base-addition salts can be obtained in a neat manner or in a suitable inert solvent by contacting the neutral form of such compounds with a sufficient amount of the desired base. Examples of pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amino or magnesium salts or similar salts. When the compounds of the present invention contain relatively basic functional groups, acid addition salts can be obtained in a neat manner or in a suitable inert solvent by contacting the neutral form of such compounds with a sufficient amount of the desired acid. Examples of pharmaceutically acceptable acid addition salts include those derived from inorganic acids such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, monohydrocarbonic acid, phosphoric acid, monohydrogenphosphoric acid, dihydrogenphosphoric acid, sulfuric acid, monohydrogensulfuric acid, hydroiodic acid or phosphorous acid and the like, and from relatively non-toxic organic acids such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, sebacic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, methanesulfonic acid and the like. Also included are salts of amino acids, such as arginine salts and the like, and salts of organic acids, such as glucuronic acid or galacturonic acid and the like (see, e.g., berge et al, "Pharmaceutical Salts", journal of Pharmaceutical Science,1977, 66, 1-19). Certain specific compounds of the invention contain basic and acidic functionalities that enable the compounds to be converted into base or acid addition salts. Supplementary active compounds are incorporated into the compositions.

Detailed Description

Mifamurtide (MTP-PE)

Mifamurtide (MTP-PE) is an artificially synthesized lipophilic Muramyl Dipeptide (MDP) analogue, namely, liposome-encapsulated muramyl tripeptide phosphatidylethylamine alcohol (N-Acetylmuramyl-alanyl-sn-glyco-3-phosphoethano lamine), which has better lipophilicity than MDP and is mainly used for treating osteosarcoma as an immune adjuvant therapy, and recently has been shown to enhance insulin sensitivity of obese mice through NOD2-IRF4 and improve glucose tolerance of high-fat diet fed mice (Cell metabolism,2017,25 (5): 1063-1074.e3).

Mifamurtide has the chemical structural formula:

pharmaceutical preparation

As known to those skilled in the art, the term "pharmaceutically acceptable carrier" as used herein includes any and all solvents, dispersion media, coatings, surfactants, antioxidants, preservatives (e.g., antibacterial, antifungal agents), isotonic agents (isotonic agents), absorption delaying agents, salts, preservatives, pharmaceutical stabilizers, binders, excipients, disintegrants, lubricants, sweeteners, flavoring agents, dyes, and the like, as well as combinations thereof (see, e.g., remington's PharmaceuticalSciences, 18 th edition, mack Printing Company,1990, pages 1289-1329). Unless any conventional carrier is incompatible with the active ingredient, its use in a therapeutic or pharmaceutical composition will be contemplated.

The therapeutic formulations of the present application may be prepared for storage in the form of tablets, capsules, granules, pellets, drop pills, oral liquids, water injections, powder injections, infusion solutions, ointments, gels or microemulsions by mixing Mifamurtide or a pharmaceutically acceptable salt, ester, hydrate thereof with an optional physiologically acceptable carrier, excipient or stabilizer (Remington's Pharmaceutical Sciences, 16 th edition, osol, A. Editions (1980)). Acceptable carriers, excipients, or stabilizers are nontoxic to recipients at the dosages and concentrations employed, and include buffers such as phosphate, citrate, histidine, and other organic acids; antioxidants (including ascorbic acid and methionine); preservatives (e.g., octadecyl benzyl dimethyl ammonium chloride, hexa methyl ammonium chloride, benzalkonium bromide, phenol, butanol or benzyl alcohol, alkyl parabens Jin Zhiru methyl or propyl parabens, catechol, resorcinol, cyclohexanol, 3-pentanol and m-cresol); a low molecular weight (less than about 10 residues) polypeptide; proteins, such as serum albumin, gelatin or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrins; chelating agents such as EDTA; sugars such as sucrose, mannose, trehalose or sorbose; salt-forming counterions such as sodium; metal complexes (e.g., zn-protein complexes) and/or nonionic surfactants such as TWEEN TM, PLURONICS TM, or polyethylene glycol (PEG).

For example, the pharmaceutical formulations of the present application may comprise excipients that maximize the permeability of the pharmaceutically active ingredient (MTP-PE) in the ileum, including, but not limited to: sodium caprate, sodium laurate, sodium palmitate, SNAC, chitosan and derivatives thereof, fatty acids, fatty acid vinegar, polyethers, bile salts, hydroxylase inhibitors, antioxidants and/or nitric oxide donors comprising nitric oxide donor groups covalently linked to various active pharmaceutical ingredients.

For example, the pharmaceutical formulations of the present application may comprise excipients that maximize the permeability of the active pharmaceutical compound in the colon, including, but not limited to: sodium caprate, sodium laurate, sodium palmitate, SNAC, chitosan and derivatives thereof, fatty acids, fatty acid vinegar, polyethers, bile salts, hydroxylase inhibitors, antioxidants and/or nitric oxide donors comprising nitric oxide donor groups covalently linked to various active pharmaceutical ingredients.

For example, the pharmaceutical formulations of the present application may contain excipients that enhance the therapeutic efficacy of the active pharmaceutical ingredient in the ileum and colon, including, but not limited to: absorption limiting agents, essential oils, natural plant extracts such as neem oil, ion exchange resins, bacterially degradable coupling linkers such as azo bonds, polysaccharides such as amylose, guar gum, pectin, chitosan, inulin, cyclodextrin, chondroitin sulfate, dextran, guar gum and locust bean gum, inhibitors of nuclear factor κb, acids such as fumaric acid, citric acid, etc., and modified forms thereof.

For example, the pharmaceutical formulations of the present application may contain excipients or other active agents or other ingredients to enhance systemic bioavailability after absorption in the small intestine, including: outflow pump inhibitors, including but not limited to PgP pump inhibitors; and metabolic inhibitors, including but not limited to cytochrome P4503A inhibitors.

For example, the pharmaceutical formulations of the present application may comprise excipients capable of reducing systemic side effects associated with small intestine absorption, including, but not limited to: antioxidants, such as curcuminoids, flavonoids, or more specifically, curcumin, beta-carotene, alpha-tocopherol, ascorbate, or lazarooid.

The pharmaceutically active ingredient (MTP-PE) can be in dissolved form or in readily soluble liquid, semi-liquid or solid form.

The pharmaceutical formulations herein may also contain more than one active compound, typically those active compounds having complementary activities that do not adversely affect each other, as desired for the particular indication being treated. Such molecules are suitably present in an amount effective for the intended purpose.

The active ingredient may also be embedded in microcapsules (e.g., hydroxymethyl cellulose microcapsules or gelatin microcapsules and poly (methyl methacrylate) microcapsules), colloidal drug delivery systems (e.g., liposomes, albumin microspheres, microemulsions, nanoparticles and nanocapsules), or emulsions, respectively, prepared, for example, by coacervation techniques or interfacial polymerization. Such techniques are disclosed in Remington's Pharmaceutical Sciences, 16 th edition, osol, a. Editions (1980).

The pharmaceutical formulation to be used for in vivo administration must be sterile. This is easily achieved by filtration through sterile filtration membranes.

Sustained release articles can be prepared. Suitable examples of sustained-release preparations include solid hydrophobic polymeric semipermeable matrices containing Mifamurtide or pharmaceutically acceptable salts, esters, hydrates thereof, which matrices are in the form of shaped articles (e.g., films or microcapsules). Examples of sustained-release matrices include polyesters, hydrogels (e.g., poly (2-hydroxyethyl methacrylate) or poly (vinyl alcohol), polylactic acid, copolymers of L-glutamic acid and gamma-ethyl-glutamate, nondegradable ethylene-vinyl acetate, degradable lactic-glycolic acid copolymers such as LUPRON DEPOTTM (injectable microspheres composed of lactic-glycolic acid copolymer and leuprorelin acetate), and poly-D- (-) -3-hydroxybutyric acid.

The pharmaceutically active ingredient (MTP-PE) of the present application may be a micro-or nano-sized particle. The pharmaceutically active ingredient may be in a soluble form. Alternatively, the pharmaceutically active ingredient may be in crystalline form or the pharmaceutically active ingredient may be in amorphous form.

The pharmaceutically active ingredient (MTP-PE) can be released along the gastrointestinal tract in a form that maximizes systemic absorption. For example, the pharmaceutically active ingredient may be released along the gastrointestinal tract in a form that maximizes lymphatic absorption. Alternatively, the pharmaceutically active ingredient may be released along the gastrointestinal tract in a form that maximizes blood brain barrier absorption. Alternatively, the pharmaceutically active ingredient may be released along the gastrointestinal tract in a form that maximizes pre-systemic absorption (pre-systemic absorption). Alternatively, the pharmaceutically active ingredient may be released along the gastrointestinal tract in a form that maximizes local gastrointestinal activity. Alternatively, the pharmaceutically active ingredient may be released along the gastrointestinal tract in a form that maximizes the activity of the gastrointestinal lumen. Alternatively, the pharmaceutically active ingredient may be released along the gastrointestinal tract in a form that maximizes chronotherapy. In all cases, the release of the active pharmaceutical ingredient is such that it is in a soluble form or readily soluble in the local GIT environment upon release.

In the present application, the pharmaceutically active ingredient (MTP-PE) may remain in the stomach area for a prolonged period of time.

In the present application, the pharmaceutically active ingredient (MTP-PE) may be in two or more forms, in the form of solid microcapsules soluble in the small intestine or in the form of pre-dissolved forms released in the colon and/or ileum.

In this application, the pharmaceutically active ingredient (MTP-PE) may be in a form that enhances dissolution and is readily absorbed upon release in the colon.

Method

Methods of treating IBD provided herein comprise administering to a patient in need of IBD treatment a composition comprising an effective amount of Mifamurtide or a pharmaceutically acceptable salt, ester, hydrate thereof.

Examples of the method of administration of the drug of the present application include oral administration and parenteral administration, and examples of the method of parenteral administration include intravascular administration (intra-arterial administration, intravenous administration, etc.), intramuscular administration, subcutaneous administration, intradermal administration, intraperitoneal administration, nasal administration, pulmonary administration, transdermal administration, etc., but are not limited thereto.

In some embodiments, the route of administration comprises oral, injection, implant, topical, spray, inhalation, or a combination thereof.

In some embodiments, the route of administration is gastrointestinal. For example, the route of administration may be oral. For another example, the route of administration may be intragastric. For another example, the route of administration may be transrectal implantation.

The dosing schedule may be determined by a user of the method, such as a prescribing physician. In one embodiment, the administration is once daily. In another embodiment, the administration is twice daily. In another embodiment, the administration is three times per day. In another embodiment, once a day, once every other day, once every third day, once every fourth day, 1 time a week, 2 times a week, 3 times a week, or 4 times a week.

In certain embodiments, mifamurtide or a pharmaceutically acceptable salt, ester, hydrate thereof is administered once every four weeks or once every 0, 2 and four weeks, followed by once every four weeks for a month period (4 weeks), or two, three, or six months or 12 months, or 18 months, or 24 months, or a lifetime administration of the patient. In certain embodiments, the treatment is self-administered by the patient.

In certain embodiments, the patient self-administers using an automatic injection device that contains a prefilled syringe. Various self-injection devices, including auto-injector devices, are known and commercially available in the art.

In certain embodiments, a near-flat dose of Mifamurtide or a pharmaceutically acceptable salt, ester, hydrate thereof is administered to a patient. A near flat dose is a specific amount of Mifamurtide or a pharmaceutically acceptable salt, ester, hydrate thereof administered to each patient, regardless of weight. Depending on the type and severity of the disease, a near flat dose of Mifamurtide between about 0.5mg and 500mg, or a pharmaceutically acceptable salt, ester, hydrate thereof, is administered to the patient, which may be one or more separate injections or infusions or administrations. Such a near flat dose may be administered subcutaneously, orally, inhaled or implanted. In certain embodiments, the near flat dose is about 1mg, or about 5mg, or about 10mg, or about 20mg, or about 50mg, or about 100mg, or about 200mg, or about 300mg, or about 400mg, or about 450mg, about 100mg, or about 200mg, or about 300mg, or about 400mg, or about 450mg.

In some embodiments of the present invention, in some embodiments, mifamurtide or a pharmaceutically acceptable salt, ester, hydrate thereof may be present in an amount of 0.01mg/Kg, 0.05mg/Kg, 0.1mg/Kg, 0.15mg/Kg, 0.2mg/Kg, 0.25mg/Kg, 0.3mg/Kg, 0.35mg/Kg, 0.4mg/Kg, 0.45mg/Kg, 0.5mg/Kg, 0.55mg/Kg, 0.6mg/Kg, 0.65mg/Kg, 0.7mg/Kg, 0.75mg/Kg, 0.8mg/Kg, 0.85mg/Kg, 0.9mg/Kg, 0.95mg/Kg, 1mg/Kg, 1.5mg/Kg, 2mg/Kg, 2.5mg/Kg, 3.5mg/Kg, 4mg/Kg, 4.5mg/Kg, 5mg/Kg, 5.5mg/Kg, 6mg/Kg, 6.5mg/Kg, 7.8 mg/Kg, 8.9 mg/Kg, 0.95mg/Kg, 1mg/Kg, 2.5mg/Kg, 3.5mg/Kg 9.5mg/Kg, 10mg/Kg, 10.5mg/Kg, 11mg/Kg, 11.5mg/Kg, 12mg/Kg, 12.5mg/Kg, 13mg/Kg, 13.5mg/Kg, 14mg/Kg, 14.5mg/Kg, 15mg/Kg, 15.5mg/Kg, 16mg/Kg, 16.5mg/Kg, 17mg/Kg, 17.5mg/Kg, 18mg/Kg, 18.5mg/Kg, 19mg/Kg, 20mg/Kg, 21mg/Kg, 22mg/Kg, 23mg/Kg, 24mg/Kg, 25mg/Kg, 26mg/Kg, 27mg/Kg, 28mg/Kg, 29mg/Kg, 30mg/Kg, 35mg/Kg, 40mg/Kg, 45mg/Kg, 50mg/Kg, 60mg/Kg, 70mg/Kg, 80mg/Kg, 90mg/Kg, 100mg/Kg, A dosage of 150mg/Kg, 200mg/Kg, 250mg/Kg, 300mg/Kg, 350mg/Kg, 400mg/Kg, 450mg/Kg, 500mg/Kg, 550mg/Kg, 600mg/Kg, 650mg/Kg, 700mg/Kg, 750mg/Kg, 800mg/Kg, 950mg/Kg, 1000mg/Kg is administered to the subject.

By following the instructions on the label, the clinical staff will be able to determine the appropriate dosage for the individual subject. The formulation and dosing regimen of the commercially available second therapeutic compound and other compounds in combination or simultaneous administration with Mifamurtide or a pharmaceutically acceptable salt, ester, hydrate thereof may be used according to the manufacturer's instructions or determined empirically by the skilled practitioner.

In this application, the terms "subject" and "patient" are used interchangeably and refer generally to mammals, such as human patients and non-human primates, as well as laboratory animals, such as rabbits, rats, and mice, among others. Animals include all vertebrates, e.g., mammals and non-mammals, such as dogs, cats, sheep, cows, pigs, rabbits, chickens, and the like. A preferred subject for practicing the methods of treatment of the invention is a human. Subjects in need of treatment include those who have had an ocular vascular disease or disorder and who are prone to develop such disorders.

In this application, the term "effective amount" or "effective dose" generally refers to an amount sufficient to achieve, or at least partially achieve, a desired effect. A "therapeutically effective amount" or "therapeutically effective dose" of a drug or therapeutic agent is generally any amount of drug that, when used alone or in combination with another therapeutic agent, promotes regression of the disease (as evidenced by a decrease in severity of symptoms of the disease, an increase in the frequency and duration of disease asymptomatic periods, or prevention of damage or disability due to the disease). "prophylactically effective amount" or "prophylactically effective dose" of a drug generally refers to an amount of a drug that inhibits the progression or recurrence of a disease when administered alone or in combination with another therapeutic agent to a subject at risk of disease progression or recurrence. The ability of a therapeutic or prophylactic agent to promote regression of a disease or inhibit the progression or recurrence of a disease can be assessed using a variety of methods known to those of skill in the art, such as in a human subject during a clinical trial, in an animal model system to predict efficacy in humans, or by assaying the activity of the agent in an in vitro assay.

In one embodiment of the combination therapy, mifamurtide or a pharmaceutically acceptable salt, ester, hydrate thereof of the present application is administered simultaneously or sequentially with one or more other active ingredients. For example, mifamurtide or a pharmaceutically acceptable salt thereof may be administered simultaneously or sequentially with another pharmaceutical composition for use as a medicament for treating IBD. The additional pharmaceutical composition may be a drug that the patient may have prescribed (e.g., an existing standard or care patent drug).

In this application, the term "treatment" generally refers to: (i) Preventing the occurrence of a disease, disorder, or condition in a patient who may be susceptible to the disease, disorder, and/or condition, but has not been diagnosed with the disease; (ii) Inhibiting the disease, disorder or condition, i.e., inhibiting its development; and (iii) alleviating the disease, disorder or condition, i.e., causing regression of the disease, disorder and/or condition and/or symptoms associated with the disease, disorder and/or condition.

In this application, the terms "comprising," "including," "having," "can," "containing," and variants thereof are intended to be open-ended transitional phrases, terms, or words that do not preclude the possibility of additional actions or structures. The singular forms "a", "an" and "the" include plural referents unless the context clearly dictates otherwise. The present disclosure also contemplates other embodiments that "comprise", "consist of" and "consist essentially of the embodiments or elements presented herein.

Except in the operating examples, or where otherwise indicated, all numbers expressing quantities of ingredients or reaction conditions used herein are to be understood as being modified in all instances by the term "about". The term "about" when used in connection with a percentage may mean ± 5%, e.g., 1%, 2%, 3% or 4%.

The application specifically discloses the following embodiments:

use of mifamurtide in the manufacture of a medicament for the treatment and/or prophylaxis of inflammatory bowel disease.

2. The use according to embodiment 1, wherein the inflammatory bowel disease comprises ulcerative colitis and crohn's disease.

3. The use of any one of embodiments 1-2, wherein the inflammatory bowel disease comprises sodium dextran sulfate (DSS) induced Inflammatory Bowel Disease (IBD).

Use of mifamurtide in the manufacture of a medicament for inhibiting secretion of an inflammatory factor in the gut.

5. The use according to embodiment 4, wherein the intestinal inflammatory factor is to include IL-1 beta, IL-1 alpha, TNF alpha, GM-CSF, IL-6, and/or IL-17.

6. The use according to any one of embodiments 1-5, wherein the main active ingredient of the medicament is Mifamurtide.

7. The use according to any one of embodiments 1-6, wherein the only active ingredient of the medicament is Mifamurtide.

8. The use according to any one of embodiments 1-7, wherein the medicament is formulated for administration by oral, inhalation, nasal spray, injection, topical, eye drops, rectal and/or vaginal.

9. The use of any one of embodiments 1-8, wherein the medicament is formulated as a solid dosage form, a solution dosage form, a nebulizer aerosol dosage form, an injection dosage form, an ointment dosage form, a skin patch dosage form, a film dosage form, a soft capsule dosage form, or a suppository dosage form.

10. The use according to any one of embodiments 1-9, wherein the medicament is formulated for administration by oral, inhalation, nasal spray, injection, topical, eye drop, rectal and/or vaginal administration and in solid dosage forms, solution dosage forms, nebulizer aerosol dosage forms, injection dosage forms, ointment dosage forms, skin patch dosage forms, film dosage forms, soft capsule dosage forms, suppository dosage forms.

11. The use according to any one of embodiments 1-10, wherein the medicament is formulated as a tablet, capsule, granule, suppository, ointment, patch, injection solution, and sustained and controlled release agent by oral, parenteral route or implantation.

12. The use according to any one of embodiments 1-11, wherein the medicament is formulated as a pulmonary or nasal inhalation aerosol, metered dose aerosol or dry powder inhaler.

13. The use according to any one of embodiments 1-12, wherein the medicament is formulated as a liquid injection or infusion.

14. A medicament for the treatment of inflammatory bowel disease comprising Mifamurtide or a pharmaceutically acceptable salt, ester, hydrate thereof, and optionally a pharmaceutically acceptable carrier.

15. The medicament of embodiment 14, wherein the inflammatory bowel disease comprises ulcerative colitis and Crohn's disease.

16. The medicament of any of embodiments 14-15, wherein the inflammatory bowel disease comprises sodium dextran sulfate (DSS) induced Inflammatory Bowel Disease (IBD).

17. The medicament of any of embodiments 14-16, wherein the medicament inhibits secretion of an inflammatory factor in the gut.

18. The medicament of any of embodiments 14-17, wherein the intestinal inflammatory factor is to include IL-1 β, IL-1 a, tnfa, GM-CSF, IL-6 and/or IL-17.

19. The medicament according to any of embodiments 14-18, wherein the main active ingredient of the medicament is Mifamurtide.

20. The medicament according to any of embodiments 14-19, whose sole active ingredient is Mifamurtide.

21. The medicament of any of embodiments 14-20, wherein the medicament is formulated for administration by oral, inhalation, nasal spray, injection, topical, eye drops, rectal and/or vaginal administration.

22. The medicament of any of embodiments 1-8, wherein the medicament is formulated as a solid dosage form, a solution dosage form, a nebulizer aerosol dosage form, an injection dosage form, an ointment dosage form, a skin patch dosage form, a film dosage form, a soft capsule dosage form, or a suppository dosage form.

23. The medicament of any of embodiments 14-22, wherein the medicament is formulated for administration by oral, inhalation, nasal spray, injection, topical, eye drop, rectal and/or vaginal administration and is in a solid dosage form, a solution dosage form, a nebulizer aerosol dosage form, an injection dosage form, an ointment dosage form, a skin patch dosage form, a film dosage form, a soft capsule dosage form, a suppository dosage form.

24. The medicament of any of embodiments 14-23 formulated as tablets, capsules, granules, suppositories, ointments, patches, water injection and sustained and controlled release agents by oral, parenteral route or implantation.

25. The medicament of any of embodiments 14-24 formulated as a pulmonary or nasal inhalation aerosol, metered dose aerosol, or dry powder inhaler.

26. The medicament of any of embodiments 14-25, formulated as a liquid injection or infusion.

27. The medicament according to any of embodiments 14-26, wherein the effective dose of Mifamurtide or a pharmaceutically acceptable salt, ester, hydrate thereof is 0.01mg/kg to 100mg/kg.

28. A method of treating inflammatory bowel disease, the method comprising administering to a subject in need thereof an effective amount of Mifamurtide.

29. The method of embodiment 28, wherein the inflammatory bowel disease comprises ulcerative colitis and crohn's disease.

30. The method of any one of embodiments 28-29, wherein the inflammatory bowel disease comprises sodium dextran sulfate (DSS) induced Inflammatory Bowel Disease (IBD).

31. A method of inhibiting secretion of an inflammatory factor in the gut, the method comprising administering to a subject in need thereof an effective amount of Mifamurtide.

32. The method of embodiment 31, wherein the intestinal inflammatory factor is to include IL-1 β, IL-1 a, tnfa, GM-CSF, IL-6, and/or IL-17.

33. The method of any one of embodiments 28-32, comprising administering Mifamurtide as the primary active ingredient to a subject.

34. The method of any one of embodiments 28-33, comprising administering Mifamurtide as the sole active ingredient to a subject.

35. The method of any one of embodiments 28-33, wherein the method further comprises administering to the subject an additional therapeutic agent.

36. The method of any one of embodiments 28-35, wherein the method comprises administration by oral, inhalation, nasal spray, injection, topical, eye drops, rectal, and/or vaginal.

Without intending to be limited by any theory, the following examples are meant to illustrate the drugs and uses of the present application, and the like, and are not intended to limit the scope of the invention of the present application.

Examples

Example 1 DSS acute enteritis model verifies the therapeutic Effect of Mifamurtide

(1) Establishment and pharmaceutical intervention of DSS enteritis

8-10 week old C57BL/6 female mice were randomly divided into groups and given different concentrations and modes of administration (gavage or intraperitoneal injection) of Mifamurtide and MDP. The experimental group drinks 3% DSS for 1-8 days and intervenes with the medicines for 1-3 days; the control group was given pure water and 1% dmso was injected intraperitoneally for 1-3 days. The experimental group lavage amount is: mifamurtide 12.5ug/g (approximately equal to 5 ug/gMDP); the intraperitoneal injection amount is as follows: mifamurtide (1 ug/g, 4ug/g, 12.5 ug/g), MDP (5 ug/g). The molding was continued for 9 days, and mice were treated on day 9.

(2) Evaluation of severity of enteritis

The evaluation index of the enteritis severity comprises weight change, colon length and pathological tissue score as follows: the body weight of the mice is monitored and recorded every day, and the stool characters and hematochezia are observed. After the molding is finished, the mice are sacrificed, colon tissues are taken out, and the length of the colon tissues is measured and recorded. Colorectal tissue was additionally HE stained. HE sections were scored for intestinal mucosal lesions and intestinal inflammation by two independent pathologists (unaware of the treatment). All scores were performed using an optimized composite scoring system, including tissue damage (0-3 points), infiltration of lamina propria inflammatory cells (0-3 points).

(3) Detection of intestinal permeability of mice with DSS enteritis

The mice in the experimental group and the control group are respectively euthanized by 600mg/kg of gastric lavage macromolecular fluorescent substance FITC-dextran after four hours, and blood is collected by heart blood sampling. After the blood is kept stand for 2 hours, the blood is centrifuged at 3000rpm for 5 minutes, serum is taken, the serum is diluted to proper concentration, the 96-well plate is added, and the multi-well detection is carried out under the condition of 485nm of excitation light and 528nm of emitted light.

(4) Serum inflammatory cytokine detection for mice with DSS enteritis

Serum samples of mice of each group were diluted and the IL-6 level in the serum of the mice was detected by a double antibody sandwich method according to the mouse-IL-6 kit instructions of proteontech.

As shown in fig. 1A-1D, different concentrations of Mifamurtide all improved DSS-induced inflammatory bowel disease. Mifamurtide at each dose was able to alleviate weight loss (FIG. 1A), improve colon length loss (FIG. 1B), reduce colon histopathological score (FIG. 1C), and decrease inflammatory factor IL-6 levels (FIG. 1D) compared to the DSS group.

As shown in fig. 2A to 2C, both intraperitoneal injection and oral administration of Mifamurtide can improve inflammatory bowel disease, and Mifamurtide in different ways has therapeutic effects. Intraperitoneal injection and oral Mifamurtide can both relieve weight loss (figure 2A), improve colon length reduction (figure 2B) and reduce colon histopathological scores (figure 2C).

As shown in fig. 3A to 3B, both Mifamurtide and MDP with equal molar mass can improve inflammatory bowel disease, and both have therapeutic effects. The intraperitoneal injection of MDP can relieve weight loss (figure 3A), reduce the damage degree of intestinal barrier (figure 3B), improve colon length reduction, reduce pathological scores of colon tissues and have better protection effect.

As shown in fig. 4A to 4C, mifamurtide has better security. After the Mifamurtide with high concentration is used, no obvious organ damage is seen compared with the control group: (FIG. 4A) liver; (fig. 4B) kidneys; (FIG. 4C) small intestine.

In conclusion, the method comprises the steps of,

mifamurtide can obviously relieve the condition of mice with DSS-induced inflammatory bowel disease, and has good improvement effects on weight, colon length reduction, intestinal barrier, serum inflammatory cytokines and histopathological scores.

2. Mifamurtide with different administration modes can improve the enteritis induced by DSS.

3. Comparing the therapeutic effect of intraperitoneal injection of Mifamurtide and MDP of equal molar mass, mifamurtide showed a trend in various aspects better than the therapeutic effect of MDP.

Mifamurtide dry prognosis high concentration experimental group small intestine, kidney and liver tissue sections all show that Mifamurtide has better safety and does not cause organ toxicity injury.

Claims

Use of mifamurtide in the manufacture of a medicament for the treatment and/or prophylaxis of inflammatory bowel disease.
2. The use according to claim 1, wherein the inflammatory bowel disease comprises ulcerative colitis and crohn's disease.
3. The use of any one of claims 1-2, wherein the inflammatory bowel disease comprises sodium dextran sulfate (DSS) induced Inflammatory Bowel Disease (IBD).
Use of mifamurtide in the manufacture of a medicament for inhibiting secretion of an inflammatory factor in the gut.
5. The use according to claim 4, wherein the intestinal inflammatory factor is to include IL-1 β, IL-1 a, tnfa, GM-CSF, IL-6 and/or IL-17.
6. The use according to any one of claims 1-5, wherein the main active ingredient of the medicament is Mifamurtide.
7. The use according to any one of claims 1-6, wherein the only active ingredient of the medicament is Mifamurtide.
8. A medicament for the treatment of inflammatory bowel disease comprising Mifamurtide or a pharmaceutically acceptable salt, ester, hydrate thereof, and optionally a pharmaceutically acceptable carrier.
9. The medicament of claim 8, wherein the inflammatory bowel disease comprises ulcerative colitis and crohn's disease.
10. The medicament of any of claims 8-9, wherein the inflammatory bowel disease comprises sodium dextran sulfate (DSS) induced Inflammatory Bowel Disease (IBD).
11. The medicament according to any of claims 8-10, wherein the medicament inhibits the secretion of intestinal inflammatory factors.
12. The medicament of claim 11, wherein the intestinal inflammatory factor is to include IL-1 β, IL-1 a, tnfa, GM-CSF, IL-6 and/or IL-17.
13. The medicament according to any of claims 8-12, whose main active ingredient is Mifamurtide.
14. The medicament according to any of claims 8-13, the only active ingredient of which is Mifamurtide.