Treatment of fibrotic diseases using metabotropic glutamate receptor 5 antagonists or / and cannabinoid receptor 1 antagonists

Abstract

The present disclosure describes methods and compositions for reducing the level or activity of metabotropic glutamate receptor 5 (mGluR5) or / and cannabinoid receptor 1 (CB1R) to treat fibrotic diseases, such as pulmonary fibrosis diseases (e.g., fibrotic interstitial lung disease). In some embodiments, an mGluR5 antagonist or / and a CB1R antagonist (e.g., a peripheral CB1R antagonist) is used to treat a fibrotic disorder, such as a pulmonary fibrosis disorder (e.g., fibrotic interstitial lung disease).

Description

[0001] Citations of Related Applications

[0002] This application claims priority to and the benefit of U.S. Provisional Application No. 63 / 352,428, filed on June 15, 2022, which is incorporated herein by reference in its entirety.

[0003] Government Support Statement

[0004] This invention was made with government support (grant number NIH Z01 AA000355-01 from the National Institute on Alcohol Abuse and Alcoholism of the National Institutes of Health). Technical Field

[0005] The present invention generally relates to compositions and methods for treating fibrotic diseases such as fibrotic lung disease (eg, fibrotic interstitial lung disease) by reducing the activity of cannabinoid receptor 1 (CB1R) or / and metabotropic glutamate receptor 5 (mGluR5). Background Art

[0006] Chronic pulmonary fibrosis is caused by scarring throughout the lungs, which can be caused by a variety of conditions, including chronic inflammatory processes (e.g., sarcoidosis and Wegener's granulomatosis), infections, environmental factors (e.g., asbestos, silica, and exposure to certain gases), exposure to ionizing radiation (e.g., radiation therapy to treat chest tumors), chronic diseases (e.g., lupus and rheumatoid arthritis), and even certain medications. In a condition called hypersensitivity pneumonitis, an increased immune response after inhalation of organic dusts or occupational chemicals can lead to pulmonary fibrosis. This condition is often caused by inhaling dust contaminated with bacteria, fungi, or animal products. In some types of pulmonary fibrosis, such as nonspecific interstitial pneumonia (NSIP), subjects may respond to immunosuppressive therapy. In many cases, there is no identifiable cause for the development of chronic lung inflammation and fibrosis, and patients with this disease often do not respond to medical treatment. This is especially true for patients with idiopathic pulmonary fibrosis (IPF). Treatment options for idiopathic pulmonary fibrosis are very limited. There is no evidence that any medication can treat this condition because scarring is permanent once it develops. For most cases of fibrotic interstitial lung disease (ILD), lung transplantation is the only available treatment option.

[0007] Research trials are currently underway using different drugs to reduce the fibrotic scar. Because some types of pulmonary fibrosis can respond to corticosteroids (such as prednisone) and / or other drugs that suppress the body's immune system, these types of drugs are sometimes prescribed in an attempt to reduce the process that leads to fibrosis. However, it is widely recognized that there are currently no truly effective treatments for fibrotic diseases. It is standard clinical practice to give patients prednisone and azathioprine, but there is no data to show that these drugs have a significant therapeutic benefit. In fact, the side effects of these drugs can cause death in patients with usual interstitial pneumonia (UIP). In addition, recently approved drugs for the treatment of idiopathic pulmonary fibrosis (such as pirfenidone and nintedanib) are not enough to completely slow the progression of fibrosis.

[0008] Therefore, there is an urgent need to identify therapeutic targets and effective therapeutic agents for the treatment of fibrotic diseases including pulmonary fibrosis (eg, fibrotic interstitial lung disease). Summary of the invention

[0009] The present disclosure addresses the need for effective drugs for treating fibrotic diseases, including fibrotic lung diseases. The present disclosure provides antagonists of metabotropic glutamate receptor 5 (mGluR5) or peripheral antagonists of cannabinoid 1 receptor (CB1R), or a combination thereof, for treating fibrotic diseases, including fibrotic lung diseases.

[0010] In one aspect, the present invention relates to a method of reducing fibrosis in a fibrotic disorder (e.g., pulmonary fibrosis), comprising reducing the activity of cannabinoid receptor 1 (CB1R) or / and metabotropic glutamate receptor 5 (mGluR5) in fibroblasts or / and fibroblasts at a fibrotic lesion. Fibrotic lesions in organs such as the lungs are areas of scarring of organ tissue such as lung tissue.

[0011] On the other hand, the present invention relates to a method for preventing, inhibiting the development, treating, improving, slowing down or alleviating one or more symptoms of a fibrotic disorder (e.g., pulmonary fibrosis) in a subject, or reversing the disorder, or otherwise achieving a therapeutic result, comprising administering to the subject a therapeutically effective amount of a composition comprising a metabotropic glutamate receptor 5 (mGluR5) antagonist or / and a therapeutically effective amount of a composition comprising a peripheral cannabinoid receptor 1 (CB1R) antagonist. In combination therapy, the mGluR5 antagonist and the peripheral CB1R antagonist may be provided in the same composition or in separate compositions.

[0012] The features and advantages of the present disclosure will be better understood by referring to the following detailed description, which sets forth illustrative embodiments of the present disclosure, and the accompanying drawings. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] The following figures are exemplary embodiments, wherein like elements are provided with like reference numerals.

[0014] Figure 1A and Figure 1B Increased mGluR5 expression and glutamate levels in bleomycin-induced pulmonary fibrosis in mice are shown. (A) Glutamate levels in the lung. (B) Expression of the Grm5 gene encoding mGluR5. Data represent mean ± SEM. n = 4 for the control group and n = 5 for the bleomycin group. Data were analyzed by T-test. *(P < 0.05) indicates statistical significance compared to the control group.

[0015] Figure 2A-2C The absence of mGluR5 is shown to protect mice from mortality and fibrosis due to bleomycin-induced pulmonary fibrosis (PF). (A) Generation of mGluR5 knockout (KO) mice and study design. (B) Survival curves. (C) Hydroxyproline levels as a marker of fibrosis. Data represent mean ± SEM. Control group n = 4, 14 days after bleomycin group n = 5, 28 days after bleomycin group n = 4. Data were analyzed by two-way ANOVA and Sidak's multiple comparison test. *(P < 0.05), **(P < 0.01), ***(P < 0.001), and ****(P < 0.0001) indicate statistical significance.

[0016] Figure 3A-3G Figures show that the absence of mGluR5 prevents the bleomycin-induced decrease in lung function. (A) Pressure-volume curve, (B) tissue elasticity, (C) peripheral airway resistance, (D) forced vital capacity, (E) forced expiratory volume (FEV), (F) inspiratory volume, (G) inspired air volume, as measures of lung function in wt (wild type) and mGluR5 KO mice in the control group and 14 and 28 days after a single dose of oropharyngeal bleomycin 1 U / kg. Data represent mean ± SEM. n = 4 in the control group, n = 4 in the group 14 days after bleomycin, and n = 6 in the group 28 days after bleomycin. Data were analyzed by two-way ANOVA and Sidak's multiple comparison test. *(P < 0.05), **(P < 0.01), ***(P < 0.001), and ****(P < 0.0001) indicate statistical significance.

[0017] Figure 4A and Figure 4B Pharmacological inhibition of mGluR5 by CTEP is shown to prevent death. (A) Experimental design, (B) Survival curves. Data represent mean ± SEM. Bleomycin + vehicle group n = 8, Bleomycin + CTEP (3 mg / kg) group n = 8. Mice were treated with vehicle or CTEP by daily oral gavage for 14 days.

[0018] Figure 5A-5C Pharmacological inhibition of mGluR5 by fenobam is shown to reduce pulmonary fibrosis and decreased lung function in the bleomycin-induced PF mouse model. (A) Experimental design, (B) Hydroxyproline levels in the lungs, (C) Forced expiratory volume. Data represent mean ± SEM. Control group n = 4, bleomycin + vehicle group n = 6, bleomycin + Fenobam (30 mg / kg) group n = 6. Mice were treated with vehicle or fenobam by daily oral gavage for 14 days. Data were analyzed by one-way ANOVA and Dunnett's multiple comparison test. * (P < 0.05) indicates that the difference is statistically significant.

[0019] Figure 6A-6C The results show that pharmacological inhibition of mGluR5 by basimglurant attenuates the development of bleomycin-induced PF in mice. (A) Survival curves, (B) % body weight, (C) lung function parameters of wt mice treated with vehicle or basimglurant (3 mg / kg, PO) after 28 days of OP-Bleo. Treatments were performed between day 8 and day 28 after bleomycin administration. ****P<0.0001 indicates statistically significant differences.

[0020] Figure 7 It is shown that loss of CB1R in mice attenuates bleomycin-induced changes in transcriptome clusters 0, 1, 2, 3, and 4, whereas loss of mGluR5 prevents changes in clusters 5 and 6. Figure 7 Left part: After PF induction with bleomycin, the total number of differentially expressed genes in CB1R knockout mice showed significantly lower changes compared with wild-type. Figure 7 Middle part: Transcriptome-wide co-expression network analysis data of the bleomycin-induced PF mouse model identified 7 different clusters, of which clusters 0, 1, and 4 were significantly upregulated after CB1R gene deletion in bleomycin-induced PF mice, while clusters 2 and 3 were downregulated. Figure 7 Right part: In bleomycin-induced PF mice, 408 out of 470 genes in clusters 5 and 6 remained unchanged after CB1R deletion but were significantly upregulated after mGluR5 gene deletion.

[0021] Figure 8 It is shown that genes belonging to clusters 5 and 6 in mouse lung are also regulated in human pulmonary fibrosis. In bleomycin-induced PF mice, 214 of the 408 genes in clusters 5 and 6 affected by mGluR5 deletion showed transcriptional profiles similar to those in advanced human IPF patients. DETAILED DESCRIPTION

[0022] Although various embodiments of the present disclosure are described herein, it will be apparent to those skilled in the art that these embodiments are provided by way of example only. Without departing from the present disclosure, it will be clear to those skilled in the art that various modifications and changes as well as variations and substitutions may be made to the embodiments described herein. It should be understood that various alternatives to the embodiments described herein may be adopted in practicing the present disclosure, including materials and methods similar or equivalent to those described herein. It should also be understood that each embodiment of the present disclosure may be optionally combined with any one or more of the other embodiments consistent with the embodiment described herein.

[0023] It should also be understood that the present disclosure appropriately encompasses analogs, derivatives, prodrugs, salts, solvates, hydrates, inclusion compounds and polymorphs of all compounds / substances disclosed herein. In certain instances of the present disclosure, "analogs", "derivatives", "prodrugs", "salts", "solvates", "hydrates", "inclusion compounds" or "polymorphs" specifically recited for compounds / substances or groups of compounds / substances should not be interpreted as intentional omission of any of these forms in other instances of the present disclosure, wherein compounds / substances or groups of compounds / substances are mentioned without reciting any of these forms.

[0024] It should also be understood that the present disclosure encompasses all possible stereoisomers of the compounds / substances described herein, including all possible diastereomers and enantiomers and racemic mixtures of enantiomers, not just specific stereoisomers as indicated by the drawn structure or nomenclature. Certain embodiments of the present disclosure relate to specific stereoisomers indicated by drawn structures or nomenclature. The specific expression "or its stereoisomers" or similar words for a compound / substance or a group of compounds / substances in certain cases of the present disclosure should not be interpreted as an intentional omission of any other possible stereoisomers of the compound / substance or group of compounds / substances in other cases of the present disclosure, wherein the compound / substance or group of compounds / substances is mentioned without the expression "or its stereoisomers" or similar words.

[0025] the term

[0026] Unless otherwise defined or otherwise clearly indicated when used in this article, all technical and scientific terms used in this article have the same meaning as those generally understood by those skilled in the art to which this application belongs. All cited patents, patent applications and other references are incorporated herein in their entirety by reference. However, if a term in this application contradicts or conflicts with a term in an incorporated reference, the term in this application takes precedence over the conflicting term in the incorporated reference.

[0027] Alternatively, compositions, methods and articles may comprise any suitable material, step or component disclosed herein, or be composed of any suitable material, step or component disclosed herein, or be composed of any suitable material, step or component disclosed herein substantially. Compositions, methods and articles may additionally or alternatively be formulated to be free of or substantially free of any material (or species), step or component that is not necessary for achieving the function or purpose of the compositions, methods and articles.

[0028] All ranges disclosed herein include the endpoints and all intermediate values ​​of the ranges, and the endpoints can be independently combined with each other (e.g., "up to 25% by weight, or more specifically, 5% by weight to 20% by weight" includes the endpoints and all intermediate values ​​of the ranges of "0% by weight to 25% by weight" and "5% by weight to 20% by weight"). The term "combination" includes blends, mixtures, alloys, reaction products, etc. "First", "second" and other terms do not indicate any order, quantity or importance, but are used to distinguish one element from another. The terms "one", "an" and "the" do not indicate a limit on quantity, and should be interpreted as covering the singular and plural, unless otherwise specified herein or clearly contradictory to the context. Unless otherwise explicitly stated, the meaning of the word "or" is "and / or". Throughout the specification, references to "some embodiments", "embodiments", etc. mean that the specific elements described in connection with the embodiment are included in at least one embodiment described herein, and may or may not exist in other embodiments. In addition, it should be understood that the described elements can be combined in any suitable manner in various embodiments. "Combinations thereof" are open ended and include any combination consisting of at least one listed component or element and optionally unlisted similar or equivalent components or elements.

[0029] “Or / and” and “and / or” mean “either…or…, or both” when referring to two elements, and mean “either…,…or…, or any combination or all of the above” when referring to three or more elements. For example, the phrase “A or / and B” means “A or B, or both A and B”, and the phrase “A, B or / and C” means “A, B or C, or any combination or all of them”.

[0030] The term "about" or "approximately" represents an acceptable error of a specific value determined by a person of ordinary skill in the art, which depends in part on how to measure or determine the value. In certain embodiments, the term "about" or "approximately" is represented within a standard deviation. In some embodiments, when a specific margin of error (e.g., the standard deviation of the mean value given in a chart or data table) is not listed, the term "about" or "approximately" means to cover the range of the value and the range included by rounding up or down to the value, taking into account significant figures. In certain embodiments, the term "about" or "approximately" represents within 10% or 5% of the specified value. Whenever the term "about" or "approximately" is located before the first numerical value in a series of two or more numerical values ​​or a series of two or more numerical ranges, the term "about" or "approximately" applies to each numerical value in the series of numerical values ​​or the series of numerical ranges.

[0031] The term “exemplary” is used herein to mean “serving as an example, instance, or illustration.” Any implementation or feature described herein as “exemplary” is not necessarily to be construed as preferred or advantageous over other implementations or features.

[0032] An "active agent" refers to a compound (including compounds disclosed herein), an element or a mixture that, when administered to a patient alone or in combination with another compound, element or mixture, directly or indirectly produces a physiological effect on a subject. Indirect physiological effects may occur through metabolites or other indirect mechanisms. An "active agent" may also enhance the effect of another active agent or make it more active. For example, a CB1R antagonist or an mGluR5 antagonist, when used in combination with another active agent, may enhance the activity of the other active agent, for example by reducing the effective dose of the other active agent.

[0033] A "pharmaceutical composition" is a composition comprising at least one active agent, such as a CB1R antagonist and / or an mGluR5 antagonist, or a pharmaceutically acceptable salt, hydrate or solvate thereof, and at least one pharmaceutically acceptable excipient or carrier. A "carrier" is an excipient or diluent, such as an aqueous and / or non-aqueous solvent system, through which the active agent is provided. A "pharmaceutically acceptable" excipient or carrier is generally safe, non-toxic, and has neither biological nor other adverse effects, and can be used for both veterinary and human pharmaceutical uses. Non-limiting examples of excipient types include liquid and solid fillers, diluents, binders, lubricants, glidants, surfactants, dispersants, disintegrants, emulsifiers, wetting agents, suspending agents, thickeners, solvents, isotonic agents, buffers, pH adjusters, absorption delaying agents, stabilizers, antioxidants, preservatives, antimicrobial agents, antibacterial agents, antifungal agents, chelating agents, adjuvants, sweeteners, flavoring agents, coloring agents, encapsulating materials and coating materials. The use of such excipients in pharmaceutical formulations is known in the art. For example, conventional vehicles and carriers include, but are not limited to, oils (e.g., vegetable oils, such as olive oil and sesame oil), aqueous solvents {e.g., saline, buffered saline (e.g., phosphate buffered saline [PBS]), and isotonic solutions (e.g., Ringer's solution)}, and organic solvents (e.g., dimethyl sulfoxide [DMSO] and alcohols [e.g., ethanol, glycerol, and propylene glycol]). Unless any conventional excipient or carrier is incompatible with the active agent (with respect to the content of the pharmaceutical composition, the term "active agent" or similar terms include prodrugs), the present disclosure includes the use of conventional excipients and carriers in formulations containing one or more active agents (e.g., CB1R antagonists or / and mGluR5 antagonists). See, e.g., Remington: The Science and Practice of Pharmacy, 21st ed., Lippincott Williams & Wilkins (Philadelphia, PA) (2005); Handbook of Pharmaceutical Excipients, 5th ed., Rowe et al., eds., Pharmaceutical Press and American Pharmaceutical Association (2005); Handbook of Pharmaceutical Additives, 3rd ed., Ash and Ash, eds., Gower Publishing Co. (2007); and Pharmaceutical Pre-formulation and Formulation, Gibson, ed., CRC Press (Boca Raton, FL) (2004). The pharmaceutical composition complies with the U.S. FDA GMP (Good Manufacturing Practice) standards for human or non-human drugs.

[0034] The term "pharmaceutically acceptable" means that the substance is generally safe, non-toxic, and does not produce any undue adverse, allergic or other untoward reactions when administered to animals such as humans.

[0035] "Pharmaceutically acceptable salts" include non-toxic acid addition salts or base addition salts of the parent compound, and also include pharmaceutically acceptable hydrates or solvates of such salts. Examples of pharmaceutically acceptable salts include, but are not limited to, mineral acid salts or organic acid salts of basic atoms or groups (such as amines); metal (such as alkali metal or alkaline earth metal) salts or organic (such as organic amine) salts of acidic groups (such as carboxylic acids); and the like. Pharmaceutically acceptable salts include conventional non-toxic salts and quaternary ammonium salts of the parent compound formed with, for example, non-toxic inorganic or organic acids. For example, conventional non-toxic acid addition salts include salts derived from inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, sulfamic acid, phosphoric acid, nitric acid and the like; and salts derived from organic acids such as acetic acid, propionic acid, succinic acid, glycolic acid, stearic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, pamoic acid, maleic acid, hydroxymaleic acid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-aminobenzoic acid, 2-acetoxybenzoic acid, fumaric acid, toluenesulfonic acid, methanesulfonic acid, ethanedisulfonic acid, oxalic acid, isethionic acid, HOOC-(CH2) n -COOH (wherein n is 0-4) etc. can be used as the example of the metal of cation and include but not limited to alkali metal (for example, lithium, sodium, potassium and cesium), alkaline earth metal (for example, magnesium, calcium and barium), aluminum and zinc. Metal cation can be provided by inorganic base (for example hydroxide, carbonate and bicarbonate). Non-limiting examples of organic amines that can be used to form base addition salts include chloroprocaine, choline, cyclohexylamine, dibenzylamine, N, N'-dibenzylethylenediamine, dicyclohexylamine, diethanolamine, ethylenediamine, N-ethylpiperidine, histidine, isopropylamine, N-methylglucamine, procaine, pyrazine, triethylamine, trimethylamine and tromethamine. Lists of other suitable salts can be found, for example, in G. Steffen Paulekuhn et al., Journal of Medicinal Chemistry 2007, 50, 6665 and in Handbook of Pharmaceutically Acceptable Salts: Properties, Selection and Use, ed. P. Heinrich Stahl and Camille G. Wermuth, Wiley-VCH, 2002.

[0036] The term "treat" includes alleviating, ameliorating, reversing or eliminating a medical condition or one or more symptoms or complications associated with the condition, as well as alleviating, ameliorating or eliminating one or more causes of the condition. "Treatment" of a medical condition includes prophylaxis, prevention, reducing the risk or likelihood of occurrence, delaying onset, reducing the incidence, frequency or severity, slowing or stopping the progression of the condition or one or more symptoms or complications associated with the condition.

[0037] In some embodiments, "treating" or "treatment" means providing an effective amount of an active agent to a subject to significantly reduce CNS disease symptoms, slow the progression of a CNS disease, or minimize the risk of developing CNS disease symptoms. In one aspect, treatment of CNS disease symptoms can be initiated before a subject develops disease symptoms.

[0038] The term "dosage regimen" refers to the dose and frequency of administration of a therapeutic agent, as well as optional duration of treatment and route of administration. Depending on the context, the term "therapeutic regimen" may refer to a dosing regimen.

[0039] The term "therapeutically effective amount" refers to an amount of a substance that, when administered to an individual, is sufficient to prevent, reduce the risk of developing, delay the onset of, or slow the progression of the disease being treated; to alleviate or ameliorate to some extent one or more symptoms or complications of the disease; or to treat a disease as defined herein. The term "therapeutically effective amount" also refers to an amount of a substance that is sufficient to elicit the biological or medical response of a cell, tissue, organ, system, animal, or human being that is being sought by a researcher, veterinarian, physician, or clinician.

[0040] In some embodiments, a "therapeutically effective amount" of a CB1R antagonist or mGluR5 antagonist is an amount effective to provide a therapeutic benefit when administered to a patient, such as reducing the number of fibroblasts at a fibrotic lesion, improving or alleviating one or more symptoms of a fibrotic disorder, or improving one or more lung function parameters, such as pressure-volume loops, tissue stiffness, peripheral airway resistance, forced vital capacity, airflow, inspiratory volume, and inspired air volume. Thus, a therapeutically effective amount of a compound is also an amount sufficient to significantly reduce the symptoms of the disease or disorder being treated. A significant reduction is statistically significant in a standard statistical significance parameter test (e.g., Student's t-test) with p<0.05.

[0041] The term "medical condition" (or simply "condition") includes diseases and disorders. The terms "disease" and "disorder" are used interchangeably herein.

[0042] "Administering" means giving, providing, applying or distributing by any suitable route. Administration of a combination of active agents includes administration in a single formulation or unit dosage form, administration of each active agent in the combination simultaneously but separately, or administration of each active agent in the combination sequentially by any suitable route. The dosage of each active agent in the combination may require more frequent administration of one of the active agents compared to the other active agents in the combination. Therefore, in order to allow for appropriate dosage, the packaged drug may include one or more dosage forms containing the combination of active agents, and one or more dosage forms containing one combination of active agents but not the other active agents in the combination.

[0043] The term "combination therapy" refers to the use of two or more therapeutic (active) drugs to treat a disease or condition. Such administration includes co-administration of therapeutic agents in a substantially simultaneous manner, such as in a single dosage form with a fixed ratio of active ingredients or in separate dosage forms of each active ingredient. In addition, such administration encompasses administration of each therapeutic agent in a sequential manner (at about the same time or at different times). In either case, the treatment regimen provides a beneficial effect of each therapeutic agent in the drug combination in treating the condition or disease.

[0044] A "patient" or "subject" is a member of any mammalian or non-mammalian species that may be in need of medical treatment. Medical treatment may include, for example, treatment of an initial or existing condition, or diagnostic treatment. Mammals include, but are not limited to, primates (e.g., humans), canines, felines, ungulates (e.g., cattle, horses, sheep, and swine [e.g., pigs]), rodents, and lagomorphs. In some embodiments, a subject or patient is a human or a non-human animal of commercial importance (e.g., livestock or domesticated animals). In certain embodiments, a patient is a human patient.

[0045] A significant change or difference is any detectable change or difference that is statistically significant in a standard parametric test of statistical significance (eg, Student's T-test with p<0.05).

[0046] Treatment of fibrotic diseases by reducing the level or activity of mGluR5 and / or CB1R

[0047] In some embodiments, the invention relates to a method of treating a fibrotic disorder in a mammal comprising reducing the activity of CB1R or mGluR5 or a combination thereof in fibrocytes and / or fibroblasts present at or associated with a fibrotic lesion in said fibrotic disorder.

[0048] In one aspect, reducing the activity of CB1R or mGluR5 is achieved by reducing the expression of the gene encoding CB1R or mGluR5. Gene expression can be reduced by reducing CNR1 (CB1R) or GRM5 (mGluR5) mRNA at the transcription stage, for example, by introducing small interfering RNA (siRNA) molecules for CB1R or mGluR5 mRNA, or by suppressing the translation of CB1R or mGluR5 mRNA at the translation stage to reduce gene expression, for example, by using microRNA designed to bind CB1R or mGluR5 mRNA. Alternatively, the expression of one or more genes related to CB1R or mGluR5 activity is reduced. For example, the expression of at least one effector gene or activator gene related to CB1R or mGluR5 activity is reduced. In another aspect, one or more active agents are locally delivered to the lungs by an aerosol, atomizer or inhaler, thereby reducing the signal transduction of lung receptors, thereby reducing CB1R or mGluR5 activity.

[0049] In another aspect, reducing the activity of CB1R or mGluR5 comprises administering to a mammal (e.g., a human) an agent (e.g., a ligand or drug) that blocks or attenuates a biological response by binding to and blocking the receptor, in an amount effective to treat a fibrotic disorder (e.g., alleviate one or more symptoms). Such drugs may be known CB1R or mGluR5 antagonists. Examples of drugs known as CB1R antagonists include, but are not limited to, peripheral CB1R antagonists, e.g., zevaquenabant (MRI-1867), MRI-1891, and TM-38837. Drugs known as mGluR5 antagonists include, but are not limited to, negative allosteric modulators of mGluR5 glutamate signaling such as fenobam, basimglurant, raseglurant, and dipraglurant, and selective mGluR5 antagonists that inhibit activation of mGluR5 receptors such as CTEP (2-chloro-4-((2,5-dimethyl-1-(4-(trifluoromethoxy)phenyl)-1H-imidazol-4-yl)ethynyl)pyridine), mavoglurant, auglurant, and remeglurant. In one aspect, the negative allosteric modulator of mGluR5 is fenobam.

[0050] In one aspect, reducing the activity of mGluR5 comprises administering mGluR5 antagonists to a subject. In one aspect, mGluR5 antagonists are negative allosteric modulators of mGluR5. In another aspect, mGluR5 antagonists are selective inhibitors of mGluR5 activation. In another aspect, mGluR5 antagonists comprise a combination of a selective inhibitor of mGluR5 activation and a negative allosteric modulator of mGluR5. In one aspect, the negative allosteric modulator of mGluR5 is any one or a combination of fenobam, basimglurant, raseglurant and diplaglurant. In some aspects, the selective inhibitor of mGluR5 activation is any one or a combination of CTEP, mavoglurant, auglurant and remeglurant.

[0051] In another aspect, a combination of an mGluR5 antagonist and a CB1R antagonist (e.g., a peripheral CB1R antagonist) is used to treat fibrotic diseases. By reducing the action / activity of CB1R and the action / activity of mGluR5, this combination can provide a higher therapeutic effect than treatment with either therapeutic agent alone. In some embodiments, the combination of an mGluR5 antagonist and a CB1R antagonist (e.g., a peripheral CB1R antagonist) has a synergistic effect in treating fibrotic diseases.

[0052] In one aspect, the fibrotic disorder of the treatment subject includes administering to the subject a conjoint therapy comprising an mGluR5 antagonist and a CB1R antagonist (e.g., a peripheral CB1R antagonist). In one aspect, treatment includes administering a peripheral CB1R antagonist and a negative allosteric modulator of mGluR5. In another aspect, treatment includes administering a peripheral CB1R antagonist and a selective inhibitor of mGluR5 activation. On the other hand, treatment includes administering a peripheral CB1R antagonist, a negative allosteric modulator of mGluR5, and a selective inhibitor of mGluR5 activation. In some aspects, the peripheral CB1R antagonist is zevaquenabant, and the negative allosteric modulator of mGluR5 is fenobam. In some aspects, the peripheral CB1R antagonist is zevaquenabant, and the negative allosteric modulator of mGluR5 is basimglurant. In some aspects, the peripheral CB1R antagonist is zevaquenabant, and the negative allosteric modulator of mGluR5 is raseglurant. In some aspects, the peripheral CB1R antagonist is zevaquenabant and the negative allosteric modulator of mGluR5 is dipraglurant. In some aspects, the peripheral CB1R antagonist is zevaquenabant and the selective inhibitor of mGluR5 activation is CTEP. In some aspects, the peripheral CB1R antagonist is zevaquenabant and the selective inhibitor of mGluR5 activation is mavoglurant. In some aspects, the peripheral CB1R antagonist is zevaquenabant and the selective inhibitor of mGluR5 activation is auglurant. In some aspects, the peripheral CB1R antagonist is zevaquenabant and the selective inhibitor of mGluR5 activation is remeglurant.

[0053] In some aspects, the use of mGluR5 antagonists or / and CB1R antagonists (e.g., peripheral CB1R antagonists) to treat fibrotic diseases in subjects includes administering to the subject a maximum effective dose of the antagonist. The maximum effective dose is generally defined as the range between the minimum effective dose (MED) and the maximum tolerated dose (MTD). The MED is defined as the lowest dose level of a drug that provides a clinically significant response in average efficacy and is also statistically significantly better than the response provided by a placebo. Similarly, the MTD is the highest possible but still tolerable dose level for pre-specified clinically limited toxicity. For example, in one aspect, treating a fibrotic disorder in a subject comprises administering to the subject a daily dose of a peripheral CB1R antagonist, such as about 1-200 mg of zevaquenabant or about 0.5-100 mg of MRI-1891 orally, or / and a daily dose of an mGluR5 antagonist, such as about 0.5-100 mg / kg of CTEP orally, about 50-700 mg of fenobam orally, about 0.5-5 mg of basimglurant orally, about 25-150 mg of dipraglurant orally, about 25-200 mg of mavoglurant or orally about 25-200 mg of auglurant. The daily dose of the peripheral CB1R antagonist or / and the mGluR5 antagonist can be taken once or in divided doses (e.g., twice or three times a day to reach the total daily dose).

[0054] In some embodiments, the present invention provides a pharmaceutical composition comprising an mGluR5 antagonist for use in treating a fibrotic disease. In another embodiment, the present invention provides a pharmaceutical composition for use in treating a fibrotic disease comprising a CB1R antagonist (e.g., a peripheral CB1R antagonist). In another embodiment, the present invention provides a pharmaceutical composition for use in treating a fibrotic disease comprising a combination of an mGluR5 antagonist and a CB1R antagonist (e.g., a peripheral CB1R antagonist).

[0055] Other embodiments provide the use of an mGluR5 antagonist or / and a CB1R antagonist (e.g., a peripheral CB1R antagonist) in the manufacture of a pharmaceutical composition for treating a fibrotic disease in a subject, and the use of an mGluR5 antagonist or a CB1R antagonist (e.g., a peripheral CB1R antagonist) in the manufacture of a pharmaceutical composition for treating a fibrotic disease in a subject when administered in combination with a CB1R antagonist (e.g., a peripheral CB1R antagonist) or an mGluR5 antagonist, respectively.

[0056] In some embodiments, a pharmaceutical composition comprising an mGluR5 antagonist and / or a CB1R antagonist (e.g., a peripheral CB1R antagonist) is provided in the form of a kit, and the kit further comprises a package insert comprising instructions for using the mGluR5 antagonist or the CB1R antagonist (e.g., a peripheral CB1R antagonist) or a combination of the mGluR5 antagonist and the CB1R antagonist (e.g., a peripheral CB1R antagonist) to treat a fibrotic disease in a subject.

[0057] In other embodiments, the fibrotic condition of the subject is treated by administering to the subject a combination of a mGluR5 antagonist or / and a CB1R antagonist (e.g., a peripheral CB1R antagonist) and one or more other therapeutic agents. In some embodiments, one or more other therapeutic agents include one or more therapeutic agents for treating fibrotic diseases, such as anti-inflammatory agents or immunosuppressants (e.g., azathioprine or corticosteroids such as prednisone), or pirfenidone or nintedanib for treating IPF or other fibrotic diseases. In further embodiments, one or more other therapeutic agents include an inhibitor of transforming growth factor β (TGF-β), a major promoter of fibrosis, or an antagonist of a TGF-β receptor.

[0058] Fibrotic diseases and fibrotic conditions that can be treated by the methods, therapeutic agents, and pharmaceutical compositions described herein include, but are not limited to, pulmonary fibrosis, chronic obstructive pulmonary disease (COPD), idiopathic pulmonary fibrosis (IPF), Hermansky-Pudlak syndrome pulmonary fibrosis (HPSPF), interstitial lung disease (including usual interstitial pneumonia [UIP] and scleroderma-related interstitial lung disease [Ssc-ILD]), respiratory bronchiolitis / interstitial lung disease, hypersensitivity pneumonitis, primary pulmonary hypertension (including prevention of plexiform lesion formation), chronic graft-versus-host disease (cGVHD), liver / liver fibrosis, cirrhosis, nonalcoholic steatohepatitis (NASH), cardiac fibrosis, myocardial fibrosis (e.g., interstitial fibrosis, subepicardial fibrosis, subendocardial fibrosis, and replacement fibrosis), cardiomyopathy, congestive heart failure, renal fibrosis, chronic kidney disease, diabetic nephropathy, retroperitoneal fibrosis (Ormond syndrome), nephrogenic systemic fibrosis, Schistosoma mansoni infection, herpes virus-related diseases (including pulmonary and skin manifestations), SARS-CoV-2 and its variants-related lung diseases, alcohol use disorder (AUD)-related lung injury, acute respiratory distress syndrome (ARDS), synthetic cannabinoid-induced lung injury and respiratory failure, keloids, lupus, nephrogenic fibrosing skin disease, fibrotic lesions associated with Schistosoma japonicum infection, autoimmune diseases (e.g., rheumatoid arthritis), pathogenic fibrosis, Lyme disease, matrix remodeling in pancreatitis, matrix fibrosis, uterine fibroids, ovarian fibrosis, corneal fibrosis, ischemia-related diseases, including pre-ischemic and post-ischemic conditions (e.g., congestive heart failure), postoperative scars (including abdominal adhesions), wide-angle glaucoma trabeculotomy, and any combination thereof. In certain embodiments, the fibrotic disorder is chronic pulmonary fibrosis.

[0059] Pulmonary fibrosis can occur in patients with a variety of diseases, such as congestive heart failure, atypical pneumonia (including Pneumocystis pneumonia), and lymphovascular spread of cancer. Environmental or occupational exposures, including inhalation of inorganic dusts (e.g., silicone, asbestos, beryllium, and black lung) are also thought to cause lung diseases characterized by pulmonary fibrosis. Pulmonary fibrosis may also develop from exposure to protein antigens (e.g., farmer's lung, pigeon breeder's lung, hot tub lung) and from exposure to toxic gases, fumes, aerosols, and vapors (e.g., silo filler's disease). Exposure to radiation, including ionizing radiation used in medical treatment, is also a well-known cause of pulmonary fibrosis. Pulmonary fibrosis may also occur in rheumatic or connective tissue diseases, such as scleroderma, rheumatoid arthritis, mixed connective tissue disease, and systemic lupus erythematosus. In addition, pulmonary fibrosis may occur in pulmonary-renal syndrome (e.g., Wegner and Goodpasture disease), sarcoidosis and other granulomatous diseases (e.g., berylliosis), systemic diseases such as hepatitis C, inflammatory bowel disease, acquired immunodeficiency syndrome, and idiopathic or rare diffuse parenchymal lung diseases (DPLDs), such as cryptogenic organizing pneumonia (COP, idiopathic), pulmonary Langerhans cell histiocytosis (rare), and eosinophilic pneumonia. In addition, pulmonary fibrosis may occur in tuberous sclerosis, neurofibromatosis, Niemann-Pick disease, Gaucher disease, and Hermansky-Pudlak syndrome.

[0060] Pulmonary fibrosis can lead to pulmonary hypertension because scar tissue compresses the pulmonary arteries, resulting in increased pulmonary artery and right ventricular pressures, which in turn leads to increased left ventricular pressures. Therefore, in another aspect, the present disclosure provides the use of mGluR5 antagonists or CB1R antagonists (e.g., peripheral CB1R antagonists) or combinations thereof to treat pulmonary hypertension. Pulmonary hypertension (PH) and PH-related diseases include, but are not limited to, functional class I to IV pulmonary hypertension, primary pulmonary hypertension (PPH), secondary pulmonary hypertension (SPH), familial PPH, sporadic PPH, precapillary pulmonary hypertension, pulmonary arterial hypertension (PAH), pulmonary venous hypertension, idiopathic pulmonary hypertension, thrombotic pulmonary arteriopathy (TPA), plexogenic pulmonary arteriopathy, and pulmonary hypertension associated with, related to, or secondary to left ventricular dysfunction, mitral valve disease, constrictive pericarditis, aortic Valvular stenosis, cardiomyopathy, mediastinal fibrosis, anomalous pulmonary venous drainage, pulmonary veno-occlusive disease, collagen vascular disease, congenital heart disease, HIV infection, exposure to drugs and toxins such as fenfluramine, chronic obstructive pulmonary disease, lung disease, interstitial lung disease, sleeping disorders or breathing affected by them, alveolar hypoventilation disorders, prolonged exposure to high altitude, neonatal lung disease, alveolar capillary dysplasia, sickle cell disease or other coagulation disorders such as chronic thromboembolism, connective tissue disease, lupus, schistosomiasis, sarcoidosis, or pulmonary capillary hemangiomatosis.

[0061] In some embodiments, the pulmonary hypertension treated with an mGluR5 antagonist or / and a CB1R antagonist (e.g., a peripheral CB1R antagonist) is pulmonary hypertension associated with respiratory disease and / or hypoxemia, including chronic obstructive pulmonary disease, interstitial lung disease, sleep disorders and breathing affected therefrom, alveolar hypoventilation disorders, long-term exposure to high altitude, neonatal lung disease, and alveolar capillary dysplasia. In certain embodiments, pulmonary hypertension is associated with chronic obstructive pulmonary disease.

[0062] In some embodiments, mGluR5 antagonists and / or CB1R antagonists (e.g., peripheral CB1R antagonists) are used in combination with one or more other therapeutic agents to treat pulmonary hypertension. In some embodiments, the one or more other therapeutic agents are selected from anticoagulants, diuretics, cardiac glycosides, calcium channel blockers, vasodilators, prostacyclin analogs, endothelin receptors (e.g., ET A or / and ET B2 ) antagonists, phosphodiesterase (e.g., PDE5) inhibitors, beta-2 agonists, antimuscarinics, endopeptidase inhibitors, lipid-lowering agents, thromboxane inhibitors, and combinations thereof.

[0063] In other embodiments, agents that reduce the level or activity of mGluR5 (e.g., mGluR5 antagonists) or / and agents that reduce the level or activity of CB1R (e.g., CB1R antagonists, such as peripheral CB1R antagonists) are used to treat fibrosis associated with, induced by, or caused in response to various cancer treatments (e.g., radiotherapy). In some embodiments, such agents are used in combination with radiotherapy to treat tumors or cancer. Radiation pneumonitis and subsequent pulmonary fibrosis are side effects of radiotherapy that hinder the efficacy of radiotherapy. In cancer treatment, radiation therapy is typically administered in doses of 20-85 Gy. However, patient studies have shown that lung toxicity in the form of pneumonitis is almost linear with increasing radiation dose. Subsequently, fibrotic lesions also occur. In some embodiments, agents that reduce the level or activity of CB1R and / or agents that reduce the level or activity of mGluR5 (e.g., in fibroblasts, fibroblasts, or / and macrophages in lung tissue) are used to treat radiation pneumonitis and / or radiation pulmonary fibrosis. An agent that reduces mGluR5 levels or activity (e.g., an mGluR5 antagonist) and / or an agent that reduces CB1R levels or activity (e.g., a CB1R antagonist, such as a peripheral CB1R antagonist) can be administered before, after, and / or concurrently with radiation therapy. In some embodiments, such agents are administered within 1, 2, 3, 4, or 5 days before and / or after radiation therapy.

[0064] Another cancer therapy that has been shown to cause severe fibrosis (including pulmonary fibrosis) is treatment with bleomycin sulfate. Bleomycin is deposited in the skin and lungs, causing fibrosis. Although it is recommended to stop administration and use corticosteroids, there is currently no effective treatment for lung damage caused by bleomycin. In some embodiments, before, after or simultaneously with the administration of bleomycin sulfate, or any combination or all of them, an agent that reduces the level or activity of mGluR5 (e.g., mGluR5 antagonists) and / or an agent that reduces the level or activity of CB1R (e.g., CB1R antagonists, such as peripheral CB1R antagonists) is administered to treat bleomycin-induced fibrosis, such as pulmonary fibrosis.

[0065] In addition to bleomycin sulfate, other drugs can also induce fibrosis and lung disease, including pulmonary fibrosis. It is known that fibrosis (e.g., pulmonary fibrosis) can be caused by a variety of drugs, including chemotherapeutic agents (e.g., bleomycin, busulfan, methotrexate, and nitrosoureas), antibiotics (e.g., nitrofurantoin), antiarrhythmics (e.g., amiodarone and tocainide), anti-inflammatory drugs (e.g., sulfasalazine), gold, penicillamine, and illicit drugs (e.g., cocaine and heroin). For example, up to 25% of cancer patients receiving nitrosourea chemotherapy drugs (e.g., carmustine, lomustine, or semustine, especially carmustine) develop premature pulmonary fibrosis within 36 months of starting nitrosourea treatment. In some embodiments, agents that reduce mGluR5 levels or activity (e.g., mGluR5 antagonists) or / and agents that reduce CB1R levels or activity (e.g., CB1R antagonists, such as peripheral CB1R antagonists) (e.g., in fibroblasts in lung tissue or / and in fibroblasts) are used to treat fibrosis (e.g., pulmonary fibrosis) caused by drugs or substances. Drugs and substances that can induce fibrosis (e.g., pulmonary fibrosis) include, but are not limited to, amphotericin B, bleomycin, bromocriptine, busulfan, carbamazepine, chlorambucil, cocaine, cyclophosphamide, phenytoin, ergotamine, flecainide, heroin, melphalan, methadone, methotrexate, methylphenidate, methyl lysergic acid, mineral oil, nitrofurantoin, nitrosoureas, procarbazine, silicone, sulfasalazine, tocainide, vinca alkaloids (e.g., vinblastine and vincristine), mitomycins, and antibacterial agents. Anti-mGluR5 agents or / and anti-CB1R agents can be administered before, simultaneously with, or / and after administration or exposure to fibrosis-inducing drugs or substances. In some embodiments, anti-mGluR5 agents or / and anti-CB1R agents are used in combination with anti-inflammatory agents or immunosuppressants (e.g., azathioprine or corticosteroids such as prednisone) to treat fibrosis (e.g., pulmonary fibrosis) caused by drugs or substances.

[0066] In some embodiments, one or more therapeutic agents described herein are administered topically to the fibrotic lesion site. In a more specific embodiment, the fibrotic lesion is located in the lungs, and one or more therapeutic agents are in local contact with the lesion. In certain embodiments, a pharmaceutical composition (e.g., liposomes or polymer nanoparticles) containing one or more therapeutic agents includes a targeting moiety to specifically localize the agent to the site of the fibrotic lesion. In some embodiments, one or more therapeutic agents are administered to the lungs by oral inhalation using an inhaler, an aerosol, or a nebulizer.

[0067] In addition, the present invention provides the use of an agent that reduces the level or activity of mGluR5 in fibroblasts and / or fibroblasts at fibrotic lesions (e.g., a mGluR5 antagonist) and / or an agent that reduces the level or activity of CB1R (e.g., a CB1R antagonist, such as a peripheral CB1R antagonist) in the preparation of a medicament for treating a fibrotic disease.

[0068] The therapeutic agents described herein, including mGluR5 antagonists and CB1R antagonists, can be administered by any suitable route to treat fibrotic diseases. In some embodiments, the therapeutic agents described herein (including mGluR5 antagonists and CB1R antagonists) are administered orally. In other embodiments, the therapeutic agents described herein (including mGluR5 antagonists and CB1R antagonists) are administered topically (e.g., skin, transdermal, mucosal, transmucosal or intranasal), injected, orally or nasally inhaled into the lungs, continuously released by a reservoir or pump, or any combination thereof. When oral administration is not suitable, other conventional routes of administration can also be used, such as parenteral (e.g., subcutaneous, intravenous, intradermal, intramuscular, intramammary, intraperitoneal or intrathecal), intraocular, retrobulbar, intrapulmonary (e.g., long-term release), aerosol, sublingual, buccal, nasal, anal, vaginal or transdermal administration, or by surgical implantation at a specific site. In combination therapy, the therapeutic agents may be administered by different routes if not provided in the same pharmaceutical composition, for example one therapeutic agent is administered by inhalation and the other therapeutic agent is administered orally.

[0069] The therapeutic agents described herein, including mGluR5 antagonists and CB1R antagonists, can be administered as pure substances, but are preferably administered as pharmaceutical compositions. Therefore, the present disclosure provides pharmaceutical compositions comprising one or more therapeutic agents described herein, such as mGluR5 antagonists or / and CB1R antagonists, or pharmaceutically acceptable salts, hydrates, solvates, polymorphs, isomers, prodrugs, etc. thereof, and at least one pharmaceutically acceptable excipient or carrier.

[0070] Pharmaceutical compositions containing one or more therapeutic agents described herein (e.g., mGluR5 antagonists or / and CB1R antagonists) may also contain one or more other agents that aid in delivering the therapeutic agent to the target tissue. For example, for transmucosal administration (e.g., administration to the lungs), the pharmaceutical composition may contain a mucolytic agent (e.g., acetylcysteine, trypsin, or ambroxol).

[0071] The formulation of the pharmaceutical composition depends in part on the selected route of administration. Appropriate salts and buffers can be added to stabilize the pharmaceutical composition and allow the composition to be absorbed at the target site. For oral administration, the pharmaceutical composition can be formulated as tablets, capsules, pills or other oral administration forms. When formulated for oral administration, the pharmaceutical composition can contain about 0.1 to about 99 % by weight (wt.%), about 25 % by weight to about 50 % by weight, or about 5 % by weight to about 75 % by weight of mGluR5 antagonists and / or CB1R antagonists, or at least about 5 % by weight of mGluR5 antagonists and / or CB1R antagonists. For parenteral administration, one or more therapeutic agents can be provided in the form of sterile aqueous solution, the solution optionally containing isotonic agents (e.g., NaCl) and buffers (e.g., sodium citrate), optionally contained in disposable prefilled syringes. The therapeutic agent can also be provided in lyophilized form, so as to be re-formulated before administration. Buffers and solutions for reconstructing the therapeutic agent can be provided to produce aqueous compositions.

[0072] In certain embodiments, the pharmaceutical composition is a unit dosage form containing about 0.1 mg to about 2000 mg, about 10 mg to about 1000 mg, about 100 mg to about 800 mg, or about 200 mg to about 600 mg of a CB1R antagonist, and / or about 0.1 mg to about 2000 mg, about 10 mg to about 1000 mg, about 100 mg to about 800 mg, or about 200 mg to about 600 mg of an mGluR5 antagonist. In other embodiments, the unit dosage form contains about 1-500 mg of a CB1R antagonist, or / and about 1-500 mg of an mGluR5 antagonist.

[0073] For the treatment of disorders described herein (e.g., fibrotic disorders), in some embodiments, the daily dose of the mGluR5 antagonist or CB1R antagonist is about 0.1-140 mg, 0.5-100 mg, or 1-50 mg per kilogram of body weight. In further embodiments, the daily dose of the mGluR5 antagonist or CB1R antagonist is about 0.5 mg-7 g, 1-500 mg, 25-500 mg, 25-200 mg, 10-100 mg, or 100-200 mg. The daily dose can be administered in a single dose or divided doses (e.g., twice or three times a day to reach a total daily dose), which may depend on the specific therapeutic agent, specific route of administration, and specific disease. The daily dose of the mGluR5 antagonist or CB1R antagonist can be administered in the form of a bolus, an infusion, or a sustained-release composition. Alternatively, it may not be necessary to take the mGluR5 antagonist or CB1R antagonist every day, which can be determined by the attending physician.

[0074] In some embodiments, treatment with a therapeutic agent described herein (e.g., an mGluR5 antagonist or a CB1R antagonist) lasts for at least about 2 weeks, a month (4 weeks), or 6 weeks. In further embodiments, treatment with a therapeutic agent described herein (e.g., an mGluR5 antagonist or a CB1R antagonist) lasts for at least about 3 months, 6 months, 1 year, 2 years, or 3 years.

[0075] It will be understood that the therapeutically effective dose of a particular therapeutic agent, the frequency of administration, and the duration of treatment may depend on a variety of factors, including the particular disease, the severity of the disease, the potency of the therapeutic agent, the route and timing of administration, the patient's age, weight, general health, sex, and diet, and the patient's response to treatment, and can be determined by the treating physician.

[0076] Agents that reduce the level or activity of mGluR5 (e.g., mGluR5 antagonists) or / and agents that reduce the level or activity of CB1R (e.g., CB1R antagonists, such as peripheral CB1R antagonists) can optionally be used in combination with one or more additional therapeutic agents to treat any of the conditions described herein, such as fibrotic diseases (e.g., pulmonary fibrosis). Fibrosis is initiated when immune cells such as macrophages release soluble factors (e.g., TGF-β) to stimulate fibroblasts, which deposit connective tissue containing collagen, which is usually a response to tissue inflammation or injury. Therefore, in some embodiments, anti-mGluR5 agents or / and anti-CB1R agents are used in combination with anti-inflammatory agents or / and immunosuppressants (e.g., azathioprine or corticosteroids such as prednisone) to treat fibrotic diseases (e.g., pulmonary fibrosis). Many immunosuppressants, including corticosteroids of the glucocorticoid class, also have anti-inflammatory properties. The anti-mGluR5 agent or / and the anti-CB1R agent and the other therapeutic agent may be administered in the same pharmaceutical composition or in separate pharmaceutical compositions, and if the latter, the administration of these agents may be simultaneous or / and sequential.

[0077] Nonsteroidal anti-inflammatory drugs (NSAIDs) include but are not limited to:

[0078] acetic acid derivatives, such as aceclofenac, bromfenac, diclofenac, etodolac, indomethacin, ketorolac, nabumetone, sulindac, sulindac sulfide, sulindac sulfone, and tolmetin;

[0079] Anthranilic acid derivatives (fenamic acid esters), such as flufenamic acid, clofenamic acid, mefenamic acid, and tolfenamic acid;

[0080] Enolic acid derivatives (oxoxicam), such as droxicam, isoxicam, lornoxicam, meloxicam, piroxicam, and tenoxicam;

[0081] Propionic acid derivatives, such as fenoprofen, flurbiprofen, ibuprofen, dexibuprofen, ketoprofen, dexketoprofen, loxoprofen, naproxen, and oxaprozin;

[0082] Salicylates, such as diflunisal, salicylic acid, acetylsalicylic acid (aspirin), choline magnesium trisalicylate, and disodium salicylate;

[0083] COX-2-selective inhibitors, such as apremilast, celecoxib, etoricoxib, firocoxib, flucoxibs (e.g., flucoxib AC), lumiracoxib, mavalacoxib, parecoxib, rofecoxib, temacoxib (JTE-522), valdecoxib, 4-O-methylhonokiol, niflumic acid, DuP-697, CG100649, GW406381, NS-398, SC-58125, benzothieno[3,2-d]pyrimidin-4-onesulfonamidethio derivatives, and COX-2 inhibitors derived from Tribulus terrestris;

[0084] Other types of NSAIDs, such as monoterpenoids (e.g., eucalyptol and phenols [e.g., carvacrol]), anilinopicolinic acid (e.g., clonixin), sulfonanilides (e.g., nimesulide), and dual inhibitors of lipoxygenase (e.g., 5-LOX) and cyclooxygenase (e.g., COX-2) (e.g., chebulic acid, lexafilone, 2-(3,4,5-trimethoxyphenyl)-4-(N-methylindol-3-yl)thiophene, and di-tert-butylphenol compounds [e.g., DTPBHZ, DTPINH, DTPNHZ, and DTPSAL]); and

[0085] Its analogs, derivatives and salts.

[0086] Immunosuppressants include, but are not limited to, interferon-β (IFN-β), glucocorticoids (see below), antimetabolites (e.g., hydroxyureas [hydroxyurea], antifolates [e.g., methotrexate], and purine analogs [e.g., azathioprine, mercaptopurine, and thioguanine]), pyrimidine synthesis inhibitors (e.g., leflunomide and teriflunomide), calcineurin inhibitors (e.g., cyclosporine [cyclosporin A], pimecrolimus, and tacrolimus), inosine-5'-monophosphate dehydrogenase (IMPDH) inhibitors (e.g., mycophenolic acid and its derivatives [e.g., mycophenolate sodium and mycophenolate mofetil]), mechanistic / mammalian target of rapamycin (mTOR) inhibitors (e.g., rapamycin [sirolimus], deforolimus [ridaforolimus], everolimus, temsirolimus, umirolimus [biolimus A9], zotarolimus and RTP-801), sphingosine-1-phosphate receptor modulators (e.g., S1PR1) (e.g., fingolimod) and serine C-palmitoyltransferase inhibitors (e.g., myriocin), and their analogs, derivatives and salts.

[0087] Glucocorticoids include, but are not limited to, cortisone-type (e.g., cortisone and its derivatives [e.g., cortisone acetate], cortisone and its derivatives [e.g., cortisone acetate, cortisone-17-acetate, cortisone-17-butyrate, cortisone-17-butyrate, and cortisone-17-valerate], prednisolone, methylprednisolone and its derivatives [e.g., methylprednisolone acetate], prednisone and thiazolinone and its derivatives [e.g., thiazolinone acetate], ... betamethasone and its derivatives [e.g., betamethasone dipropionate, betamethasone sodium phosphate, and betamethasone valerate], dexamethasone and its derivatives [e.g., dexamethasone sodium phosphate], and fluocortolone and its derivatives [e.g., fluocortolone hexanoate and fluocortolone pivalate]), halogenated steroids (e.g., alclomethasone and its derivatives [e.g., alclomethasone dipropionate], beclomethasone and its derivatives [e.g., beclomethasone dipropionate, esters], clobetasol and its derivatives [e.g., clobetasol-17-propionate], clobetasol and its derivatives [e.g., clobetasone-17-butyrate], desoximetasone and its derivatives [e.g., desoximetasone acetate], diflorasone and its derivatives [e.g., diflorasone diacetate], diflucortolone and its derivatives [e.g., diflucortolone valerate], flupredniidine and its derivatives [e.g., flupredniidine acetate], fluticasone and its derivatives [e.g., fluticasone propionate [e.g., halometasol, halometasol, and its derivatives [e.g., halometasol propionate], halometasone and its derivatives [e.g., halometasone acetate], mometasone and its derivatives [e.g., mometasone furoate]), acetones and related substances (e.g., amcinonide, budesonide, ciclesonide, desonide, fluocinolone, fluocinolone, fludrocortone [fludrocortone or flurandrenolide], halcinonide, triamcinolone acetonide, and triamcinolone acetonide alcohol), carbonates (e.g., prednicarbate), and their analogs, derivatives, and salts.

[0088] The therapeutic agents described herein, including mGluR5 antagonists and CB1R antagonists, and pharmaceutical compositions containing them, can be provided in the form of a kit. When used in combination, two or more therapeutic agents, such as mGluR5 antagonists and CB1R antagonists, can be provided in the same pharmaceutical composition or in separate pharmaceutical compositions. The kit includes a package insert containing instructions for administering the therapeutic agent or pharmaceutical composition to treat any condition described herein (e.g., a fibrotic condition). If not provided in an oral dosage form such as a tablet or capsule, the therapeutic agent (e.g., mGluR5 antagonist or / and CB1R antagonist) can be provided in the form of a prefilled syringe or a vial equipped with a syringe and a needle for parenteral administration, or provided in a formulation suitable for oral inhalation, in which case the kit can include an inhaler (e.g., a metered dose inhaler, a dry powder inhaler, a soft mist inhaler, a smart inhaler, or a nebulizer).

[0089] The present disclosure is further illustrated by the following non-limiting examples.

[0090] Example

[0091] Materials and methods:

[0092] Chemicals

[0093] CTEP, fenobam, and basimglurant were purchased from MedChem Express (New Jersey, USA). Pharmaceutical grade bleomycin was provided by Hospira (Lake Forest, IL, USA). All other chemicals were provided by Sigma-Aldrich (St. Louis, MO, USA).

[0094] Experimental drug treatment

[0095] Compounds were administered once daily by oral gavage as indicated. Vehicle was DMSO: 80: saline, the ratio is 1:1:18. The oral dosage form concentrations are 0.3 and 3 mg / mL, achieving doses of 3 and 30 mg / kg, respectively.

[0096] animal

[0097] All animal procedures were performed in accordance with the rules and regulations of the Institutional Animal Care and Use Committee of the National Institute on Alcohol Abuse and Alcoholism (NIAAA) and according to the LPS-GK1 protocol. Thirteen-week-old male C57BL / 6J mice were obtained from the Jackson Laboratory (Bar Harbor, ME, USA). Grm5 - / - (mGluR5 KO) mice (JAX 003121) were purchased from Jackson Laboratory. - / -(CB1R KO) mice were generated as described in Zimmer et al., Proc. Natl. Acad. Sci. USA, 96:5780-5785 (1999). - / - and Grm5 - / - Mice have a C57BL / 6J genetic background. Grm5 - / - and Cnr1 - / - Mouse colonies were maintained by heterozygous breeding. Mice were housed individually under a 12-h light / dark cycle and fed a standard diet (Teklad NIH-31; Envigo, Huntington, UK) ad libitum.

[0098] Oropharyngeal bleomycin inhalation

[0099] According to Park et al. in Am. J. Respir. Cell Mol. Biol., 62 (2): 178-190 (2020), bleomycin was delivered by oropharyngeal inhalation to establish a bleomycin-induced pulmonary fibrosis model. In short, bleomycin was delivered to mice anesthetized with ketamine / xylazine via the oropharynx at a dose of 1 U / kg, and a sterile 100 μL pipette was used during inspiration to deliver a volume of 100 μL / 50 g body weight. Sterile saline was used as a carrier and applied to the control group. The animals were then allowed to recover from anesthesia.

[0100] Mouse survival analysis

[0101] Survival curves were plotted using GraphPad Prism 9 software. Survival rates were analyzed using the Log-rank (Mantel-Cox) test to assess statistical differences between animal groups.

[0102] Pulmonary function measurements in Xanthium sibiricum mice

[0103] Respiratory system mechanics were measured using the FlexiVent FX system (SCIREQ Inc., Montreal, Canada), which was equipped with an FX1 module and a mouse negative pressure forced exhalation (NPFE) extension device. The system was operated using FlexiWare v7.2 software. Forced oscillation techniques and forced exhalation measurements were performed as described in McGovern et al., J. Vis. Exp., 75: e50172 (2013) and Devos et al., Respir. Res., 18: 123 (2017). Pulmonary function measurements were performed at the end of the study as a terminal procedure. Mice were anesthetized by intraperitoneal (ip) injection of ketamine / xylazine, and then an 18-gauge metal cannula was inserted into the trachea through a small incision. Pancuronium bromide (0.8 mg / kg) was then administered by intraperitoneal injection to induce paralysis, and then the mice were connected to the FlexiVent and ventilation was initiated. The pressure-volume (PV) curve, airway resistance, tissue damping (G), tissue elasticity (H), forced expiratory volume per 0.1 second (FEV 0.1), forced vital capacity (FVC), inspiratory volume (IC), and inspired air volume (A) were measured. Mouse tissues were collected after lung function tests.

[0104] Hydroxyproline (Hyp) measurement

[0105] The extent of pulmonary fibrosis was biochemically quantified by measuring the hydroxyproline content of lung extracts using liquid chromatography / tandem mass spectrometry (LC-MS / MS) as described by Park et al., Am. J. Respir. Cell Mol. Biol., 62(2): 178-190 (2020).

[0106] Glutamate (L-glutamate) levels in the lung were measured by LC-MS / MS.

[0107] Lung tissue was homogenized in 600 μL of ice-cold 0.1N perchloric acid (PCA) containing 2% EDTA and 1% ethanol using a Precellys tissue homogenizer. The homogenized sample was vortexed and centrifuged at 10,000 g for 10 minutes, and then 10 μL of the hydrolyzate was diluted 100-fold by adding 990 μL of 0.1N PCA. LC-MS / MS analysis was performed using an Agilent 6470 triple quadrupole mass spectrometer coupled to an Agilent 1260 LC system. Glutamate was separated at 40°C using an Intrada amino acid column (50 mm×3 mm column, 3 μm; Imtakt). Mobile phase A contained 9 / 75 / 16 / 0.3 acetonitrile / tetrahydrofuran / 25 mM ammonium carbonate / formic acid (vol / vol / vol / vol). Mobile phase B contained 20 / 80 acetonitrile / 100 mM ammonium formate (vol / vol). Gradient elution (600 μL min -1), B was 0% for 3 min, then linearly increased to 17% B in 6.5 min, gradually increased to 100% B and maintained for 10 min, then linearly decreased to 0% B in 11 min and maintained for 13 min. The mass spectrometer was set to Agilent jet stream ionization source and operated in positive ion mode. The source parameters were capillary voltage 3500 V, gas temperature 300 °C, sheath gas temperature 25 °C, sheath gas flow rate 10 L min -1 、Gas flow rate 5L min -1 , using nitrogen as the nebulizing gas. Collision induced dissociation (CID) was performed using nitrogen. Glutamate levels were analyzed by multiple reaction monitoring. The molecular ion and fragmentation of glutamate were measured as follows: m / z 148.1→77.2 and 148.1→39.2 (CID energies were 40 V and 80 V, respectively). Glutamate levels were determined according to the standard curve using L-glutamate as the standard (Sigma). Values ​​are in nmol mg -1 Wet tissue indicated.

[0108] Real-time PCR analysis

[0109] RNA extraction was performed using RNeasy Mini Kits from Qiagen (Valencia, CA, USA). One microgram of total RNA was reverse transcribed into cDNA using the Bio-Rad iScript cDNA Synthesis Kit (Hercules, CA). Expression of target genes was quantified using a QuantStudio 3 real-time PCR instrument from Applied Biosystems using gene-specific primers and PowerSYBRGreen master mix. Pre-designed mouse Tbp (QT00198443), Grm5 (QT00288596) were used. The housekeeping gene TATA-Box binding protein (Tbp) was used as a loading control. Gene expression values ​​were calculated according to the ΔΔCt method.

[0110] Transcriptomic analysis

[0111] RNA was isolated from control and fibrotic lung tissues of wt, mGluR5 KO and CB1R KO mice. RNA sequencing was then performed by Azenta Genewiz (New Jersey, USA) on an Illumina NovaSeq6000 using Poly-A library selection. The raw RNA sequencing results were quantified using Kallisto [Bray et al., Nat. Biotechnol., 34(5): 525-527 (2016)] and the Ensembl mouse reference genome (GRCm39) [Zerbino et al., Nucleic Acids Res., 46: D754-D761 (2018)]. Quantified data were used for differential expression analysis using DeSeq2 [Love et al., Genome Biol., 15:550 (2014)], and co-expression networks were generated using previously described methods [Arif et al., eLife, 10:e66921 (2021)], using the top 10% of correlation results.

[0112] Example 1. Effects of knockout and antagonism of mGluR5 on bleomycin-induced mouse pulmonary fibrosis model

[0113] As described by Park et al. in Am. J. Respir. Cell Mol. Biol., 62(2): 178-190 (2020), mice showed significant pulmonary fibrosis 14 days after a single oropharyngeal instillation of bleomycin (1U / kg). Therefore, we used the 14-day time point after bleomycin treatment to study pulmonary fibrosis in mice. As seen in human idiopathic pulmonary fibrosis (IPF) [Zhao et al., BMJ Open Respir. Res., 4: e000183 (2017)], glutamate levels in mouse fibrotic lungs were significantly increased ( Figure 1A ). Recent studies have shown that alcohol-induced increases in liver glutamate levels contribute to the pathology of alcoholic liver disease (ALD) by activating metabotropic glutamate receptor 5 (mGluR5) [Choi et al., Cell Metab., 30:877-889 (2019)]. To investigate the expression level of Grm5 encoding mGluR5 in pulmonary fibrosis, quantitative PCR (qPCR) was performed. mGluR5 expression was significantly increased in mouse fibrotic lungs ( Figure 1B Since both glutamate levels and mGluR5 gene expression were significantly increased, we used mGluR5 knockout (KO) mice and wild-type (wt) littermates in a bleomycin-induced pulmonary fibrosis model to explore the potential pathological role of mGluR5 in pulmonary fibrosis by a loss-of-function approach.

[0114] Pulmonary fibrosis development and lung function were assessed in wt and mGluR5 KO mice 14 and 28 days after a single oropharyngeal injection of bleomycin (1 U / kg). Figure 2A The absence of mGluR5 significantly increased the survival rate, from 66% for wild-type mice to 100% for mGluR5 KO mice ( Figure 2B Importantly, compared with wild-type littermates, loss of mGluR5 prevented the development of lung fibrosis as measured by lung levels of hydroxyproline, a biochemical marker of fibrosis ( Figure 2C Clinically, the only marker for the progression of pulmonary fibrosis is pulmonary function testing (PFT). Therefore, any potential therapeutic target that modulates pulmonary function parameters is of great interest for identifying therapeutic targets for pulmonary fibrosis (PF). Therefore, pulmonary function parameters were measured in mice using FlexiVent (SCIREQ). The absence of mGluR5 prevented the bleomycin-induced decrease in pulmonary function parameters, such as pressure-volume loop ( Figure 3A )、Tissue hardness( Figure 3B ), peripheral airway resistance ( Figure 3C ), forced vital capacity ( Figure 3D )、Air flow( Figure 3E )、Inspiratory volume( Figure 3F ) and the amount of air intake ( Figure 3G ). These results suggest that overactive mGluR5 contributes to the development of pulmonary fibrosis.

[0115] The therapeutic potential of mGluR5 antagonists in PF was evaluated. The efficacy of CTEP, a selective inhibitor of mGluR5 activation, was evaluated in a mouse model of bleomycin-induced PF. CTEP was administered orally at a dose of 3 mg / kg daily for 28 consecutive days ( Figure 4A ) (which is the maximal effective dose for mGluR5 antagonism [Lindemann et al., J. Pharmacol. Exp. Ther., 339:474-486 (2011)]), significantly improved survival, with 100% survival in CTEP-treated mice compared to 50% survival in vehicle-treated mice ( Figure 4B ).

[0116] The negative allosteric modulator of mGluR5, fenobam, was tested in the bleomycin-induced PF mouse model. Fenobam was administered orally at a dose of 30 mg / kg daily ( Figure 5A ) (which is the maximal effective dose for inhibition of mGluR5 [Porter et al., J. Pharmacol. Exp. Ther. 315:711-721 (2005)]), for 14 days, significantly attenuated the decline in fibrosis and lung function parameters, as measured by hydroxyproline levels ( Figure 5B ) and forced expiratory volume (FEV) at 0.1 seconds (airflow) ( Figure 5C ) to measure.

[0117] Another mGluR5 negative allosteric modulator, basimglurant, was tested. Basimglurant treatment was initiated once fibrosis had developed, 7 days after bleomycin administration (Figure 6). Oral administration of 3 mg / kg basimglurant 8 to 28 days after a single bleomycin injection significantly attenuated bleomycin-induced PF, as measured by increased survival ( Fig. 6A ), weight loss ( Figure 6B ) and decreased lung tissue stiffness ( Figure 6C ).

[0118] In conclusion, in a mouse model of pulmonary fibrosis (PF), metabotropic glutamate receptor 5 (mGluR5) expression and the levels of its endogenous agonist glutamate were significantly increased in fibrotic lungs, indicating that hyperactive mGluR5 signaling in the lung contributes to PF pathology. Both genetic deletion of mGluR5 and pharmacological inhibition of mGluR5 significantly attenuated bleomycin-induced PF development and significantly improved survival. Therefore, fibrotic lung diseases could be treated through mGluR5 antagonism.

[0119] Example 2. Determination of a combined treatment method for pulmonary fibrosis

[0120] A systems biology approach was used to identify rational combination therapeutic approaches for pulmonary fibrosis. Using a bleomycin-induced pulmonary fibrosis (PF) mouse model, transcriptomic changes in mouse PF were investigated by lung transcriptomics in healthy control lungs and fibrotic lungs, which were collected 14 days after a single oropharyngeal dose of bleomycin (1 U / kg). Seven differentially expressed gene network clusters (clusters 0-6) in fibrotic mouse lungs were identified by transcriptomic analysis ( Figure 7Middle section). The goal of rational combination therapy is to alleviate the pathological changes of most or all identified network clusters in the lung transcriptome. Antagonism of cannabinoid receptor 1 (CB1R) has previously been identified as a potential approach for treating different forms of PF [Cinar et al., JCI Insight, 2(8):e92281(2017); Bronova et al., Am. J. Respir. Cell Mol. Biol., 53(4):555-562(2015); and Cinar et al., Clin. Transl. Med., 11:e471(2021)]. CB1R antagonism has also been considered as a potential approach for treating liver, kidney, heart, lung and skin fibrosis [Cinar et al., Pharmacol. Ther., 208:107477(2020)]. CB1R-mediated transcriptome changes in the bleomycin-induced PF mouse model were studied using CB1R knockout mice. Loss of CB1R attenuated the dysregulation of five of the seven gene network clusters (clusters 0, 1, 2, 3, and 4) and attenuated the development of fibrosis ( Figure 7 middle part).

[0121] Furthermore, we investigated the transcriptome changes mediated by mGluR5 in the bleomycin-induced PF mouse model by comparing wt and mGluR5 KO mice. Complementary to CB1R antagonism, loss of mGluR5 led to the normalization of the other two clusters (clusters 5 and 6) ( Figure 7 on the right).

[0122] To assess the translational value of these findings, we investigated publicly available RNAseq datasets to determine whether similar transcriptomic changes seen in mouse fibrotic lungs are also present in human IPF. Cluster 5 and 6 genes were also downregulated in human IPF compared to healthy lungs ( Figure 8 ), while deletion of CB1R in mice did not alter this. However, deletion of mGluR5 normalized the pathological transcriptomic changes in mice ( Figure 8 ).

[0123] In conclusion, the combination of a mGluR5 antagonist and a CB1R antagonist (eg, a peripheral CB1R antagonist) is identified as a method for treating fibrotic lung disease because the combination attenuated all major pathological changes in the fibrotic lung transcriptome.

[0124] It should be understood that although specific embodiments have been shown and described, various modifications may be made thereto and are contemplated herein. It should also be understood that the present disclosure is not limited to the specific examples provided herein. The description and illustration of the embodiments and examples of the present disclosure herein are not intended to be interpreted in a limiting sense. It should also be understood that all aspects of the present disclosure are not limited to the specific descriptions, configurations, or relative proportions described herein, which may depend on a variety of conditions and variables. Various modifications and changes in the form and details of the embodiments and examples of the present disclosure will be apparent to those skilled in the art. Therefore, it is contemplated that the present disclosure also encompasses any and all such modifications, changes, and equivalents.

Claims

1. A method of reducing fibrosis in a subject with a pulmonary fibrotic disease, comprising reducing the fibrotic activity of a fibrotic lesion in the subject by reducing the activity of metabotropic glutamate receptor 5 (mGluR5).

2. The method according to claim 1, wherein the activity of mGluR5 is reduced by inhibiting the expression of the mGluR5 gene.

3. The method according to claim 1, wherein the activity of mGluR5 is reduced by inhibiting the production of mGluR5 at the protein stage.

4. The method of claim 1, wherein mGluR5 activity is reduced by pharmacologically inhibiting mGluR5.

5. The method of claim 4, wherein the pharmacological inhibition of mGluR5 is achieved by administering to the subject an mGluR5 antagonist.

6. The method of claim 5, wherein the mGluR5 antagonist is a negative allosteric modulator of mGluR5 glutamate signaling or a selective mGluR5 activation inhibitor.

7. The method of claim 6, wherein the negative allosteric modulator of mGluR5 glutamate signaling is fenobam, basimglurant, raseglurant or dipraglurant, or a pharmaceutically acceptable salt, polymorph, isomer or prodrug thereof.

8. The method of claim 6, wherein the selective mGluR5 activation inhibitor is CTEP (2-chloro-4-((2,5-dimethyl-1-(4-(trifluoromethoxy)phenyl)-1H-imidazol-4-yl)ethynyl)pyridine), mavoglurant, auglurant or remeglurant, or a pharmaceutically acceptable salt, polymorph, isomer or prodrug thereof.

9. The method of any one of claims 5 to 8, further comprising administering to the subject a peripheral CB1R antagonist, wherein the peripheral CB1R antagonist is optionally zevaquenabant (MRI-1867), MRI-1891 or TM-38837, or a pharmaceutically acceptable salt, polymorph, isomer or prodrug thereof.

10. A method for treating a fibrotic disease in a subject, comprising administering to a subject in need of treatment a therapeutically effective amount of a mGluR5 antagonist in combination with a therapeutically effective amount of a CB1R antagonist, wherein the mGluR5 antagonist is a negative allosteric modulator of mGluR5 glutamate signaling or a selective mGluR5 activation inhibitor, and wherein the CB1R antagonist is a peripheral CB1R antagonist, or a pharmaceutically acceptable salt, polymorph, isomer or prodrug thereof.

11. The method according to claim 10, wherein: The negative allosteric modulator of mGluR5 glutamate signaling is fenobam, basimglurant, raseglurant or dipraglurant; The selective mGluR5 activation inhibitor is CTEP (2-chloro-4-((2,5-dimethyl-1-(4-(trifluoromethoxy)phenyl)-1H-imidazol-4-yl)ethynyl)pyridine), mavoglurant, auglurant or remeglurant; and The peripheral CB1R antagonist is zevaquenabant (MRI-1867), MRI-1891 or TM-38837; or Pharmaceutically acceptable salts, polymorphs, isomers or prodrugs thereof.

12. The method of any one of claims 1 to 11, wherein the pulmonary fibrosis disease or fibrotic disease is selected from pulmonary fibrosis, chronic obstructive pulmonary disease (COPD), idiopathic pulmonary fibrosis (IPF), Hermansky-Pudlak syndrome pulmonary fibrosis (HPSPF), interstitial lung disease (including common interstitial pneumonia and scleroderma-related interstitial lung disease [Ssc-ILD]), respiratory bronchiolitis / interstitial lung disease, hypersensitivity pneumonitis, primary pulmonary hypertension (including prevention of plexiform lesions), chronic graft-versus-host disease (cGVHD), liver / liver fibrosis, cirrhosis, non-alcoholic steatohepatitis (NASH), cardiac fibrosis, myocardial fibrosis (e.g., interstitial fibrosis, subepicardial fibrosis, subendocardial fibrosis, and replacement fibrosis), cardiomyopathy, congestive heart failure, renal fibrosis, chronic kidney disease, diabetic nephropathy, retroperitoneal fibrosis (Ormond's disease), nephrogenic systemic fibrosis, Schistosoma mansoni infection, herpesvirus-associated diseases (including pulmonary and skin manifestations), SARS-CoV-2 and its variants-associated lung disease, alcohol use disorder (AUD)-associated lung injury, acute respiratory distress syndrome (ARDS), synthetic cannabinoid-induced lung injury and respiratory failure, keloids, lupus, nephrogenic fibrosing skin disease, fibrotic lesions associated with Schistosoma japonicum infection, autoimmune diseases (e.g., rheumatoid arthritis), pathogenic fibrosis, Lyme disease, stromal remodeling in pancreatitis, stromal fibrosis, uterine fibroids, ovarian fibrosis, corneal fibrosis, ischemia-related diseases, including pre- and post-ischemic diseases (e.g., congestive heart failure), postoperative scarring (including abdominal adhesions), and wide-angle glaucoma trabeculotomy.

13. The method according to any one of claims 5 to 12, wherein the mGluR5 antagonist or / and the peripheral CB1R antagonist is administered at a daily dose of about 0.01 mg / kg to 100 mg / kg, 0.1 mg / kg to 50 mg / kg, 0.1 mg / kg to 20 mg / kg, 0.1 mg / kg to 10 mg / kg, 0.1 mg / kg to 100 mg / kg, 1 mg / kg to 100 mg / kg or 10 mg / kg to 100 mg / kg.

14. The method according to any one of claims 5 to 13, wherein the mGluR5 antagonist and / or the peripheral CB1R antagonist is administered orally, parenterally (eg, intravenously, subcutaneously or intramuscularly), intranasally, sublingually or buccally, or by oral or nasal inhalation.

15. The method according to any one of claims 1 to 14, wherein the subject is a mammal, such as a human.

16. A pharmaceutical composition comprising: (i) peripheral CB1R antagonists, and (ii) mGluR5 antagonists, and (iii) a pharmaceutically acceptable excipient or carrier.

17. The pharmaceutical composition of claim 16, wherein the pharmaceutical composition is effective for treating a fibrotic disease when administered to a subject in need thereof.

18. The pharmaceutical composition according to claim 16 or 17, wherein the pharmaceutical composition is administered orally, parenterally (e.g., intravenously, subcutaneously, or intramuscularly), intranasally, sublingually, or buccally, or by oral or nasal inhalation.

19. A pharmaceutical composition according to any one of claims 16 to 18, formulated for administration to a mammal.

20. The pharmaceutical composition of claim 19, wherein the mammal is a human.

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