Method for treating obesity using a luteinizing hormone receptor agonist

Abstract

The present disclosure is directed to methods of treating or preventing obesity using luteinizing hormone receptor agonists. More particularly, provided herein are methods of treating obesity in a subject in need thereof, comprising the step of administering a thieno[2,3-d]pyrimidine derivative to the subject in need thereof.

Description

Technical Field

[0001] Priority Claim This application claims the benefit of U.S. Provisional Application No. 63 / 366,060, filed on June 8, 2022, the contents of which are hereby incorporated by reference in their entirety.

[0002] Federally Sponsored Research or Development This invention was made with government support under Grant No. U19 AG060917 awarded by the National Institute on Aging, National Institutes of Health. The government has certain rights in the invention.

[0003] This disclosure is directed to methods of treating or preventing obesity using luteinizing hormone receptor agonists. More particularly, this disclosure is directed to methods of treating or preventing obesity using luteinizing hormone receptor agonists such as ORG 43553 and ORG 43902.

Background Art

[0004] Obesity is a complex disease with an excessive amount of body fat. Obesity can increase the risk of other diseases and health problems, including heart disease, diabetes, high blood pressure, and certain cancers. Obesity can occur at any age, but there may be hormonal and lifestyle changes that increase the risk of obesity as people get older. Additionally, muscle mass typically decreases with aging, leading to a decline in metabolism. Such changes can reduce the calories needed and increase the risk of obesity.

[0005] Luteinizing hormone (LH) and human chorionic gonadotropin (hCG) promote mammalian reproduction and support pregnancy. Reproductive cessation coincides with the menopausal transition, which is associated with an early increase in gonadotropin levels and is accompanied by a greater change in serum follicle-stimulating hormone (FSH) than in luteinizing hormone (LH). This transition is accompanied not only by rapid bone loss but also by visceral obesity, disrupted energy balance, and decreased physical activity. During menopause, in response to ovarian aging, the frequency and amplitude of the mid-cycle LH surge decrease, and visceral fat increases from about 5-8% to about 15-20% of total body fat. Epidemiological studies have also shown that in menopausal women and women with polycystic ovary syndrome (PCOS), the body mass index (BMI) is negatively correlated with serum LH levels. Therefore, there is a need for compositions and methods for treating obesity, particularly in menopausal and postmenopausal women.

Prior Art Documents

Non-Patent Documents

[0006]

Non-Patent Document 1

Non-Patent Document 2

Summary of the Invention

Problems to be Solved by the Invention

[0007] Disclosed is a method for treating obesity in a subject in need thereof, comprising administering a luteinizing hormone (LH) and human chorionic gonadotropin (hCG) receptor agonist (LH / CG receptor) to the subject. The present disclosure is based, at least in part, on the following findings: (1) Transcripts and proteins of the LH / CG receptor (LHCGR) are abundantly expressed in adipose tissue, and the levels of gonadal white adipose tissue in female mice are close to those of the ovaries themselves; (2) Activation of LHCGR by the small molecule agonist ORG 43553, which has undergone clinical trials for LH, hCG, and infertility, results in less natural fat gain in mice fed a high-fat diet, independent of testosterone; and (3) ORG 43553 reduces adipocyte differentiation and induces thermogenesis in organoid cultures, both in vitro and in vivo. In one embodiment, the LH / CG receptor agonist is ORG 43533 and / or ORG 43902. In one embodiment, ORG 43533 and / or ORG 43902 are administered at a dosage between 1 and 100 mg / kg. In one embodiment, ORG 43533 and / or ORG 43902 are provided as a pharmaceutical composition comprising a pharmaceutically acceptable carrier.

Means for Solving the Problems

[0008] In one aspect, the present disclosure provides a compound of general formula I:

[0009]

Chemical Formula

[0010] Or a pharmaceutically acceptable salt thereof (wherein R1 and R2, together with the nitrogen atom to which they are attached, optionally contain one or more heteroatoms selected from N, O and / or S and form a ring having 2 to 6 carbon atoms), comprising the step of administering a thieno[2,3-d]pyrimidine derivative to a subject in need thereof, to provide a method for treating obesity in a subject in need thereof. In some embodiments, the thieno[2,3-d]pyrimidine derivative is tert-butyl 5-amino-2-methylthio-4-(3-(2-(azetidin-1-yl)-acetamido)-phenyl)-thieno[2,3-d]pyrimidine-6-carboxamide; tert-butyl 5-amino-2-methylthio-4-(3-(2-(morpholin-4-yl)-acetamido)-phenyl)-thieno[2,3-d]pyrimidine-6-carboxamide; tert-butyl 5-amino-2-methylthio-4-(3-(2-(thiomorpholin-4-yl)-acetamido)-phenyl)-thieno[2,3-d]pyrimidine-6-carboxamide; tert-butyl 5-amino-2-methylthio-4-(3-(2-(piperidin-1-yl)-acetamido)-phenyl)-thieno[2,3-d]pyrimidine-6-carboxamide; tert-butyl 5-amino-2-methylthio-4-(3-(2-(pyrrolidin-1-yl)-acetamido)-phenyl)-thieno[2,3-d]pyrimidine-6-carboxamide or tert-butyl 5-amino-2-methylthio-4-(3-(2-(piperazin-1-yl)-acetamido)-phenyl)-thieno[2,3-d]pyrimidine-6-carboxamide, a compound selected from the group consisting of. In some embodiments, the thieno[2,3-d]pyrimidine derivative is ORG 43588.

[0011] It should be noted that there may be a typo in the original text where "ORG 43588" is likely "ORG 43533". The translation has been made according to the provided text.In some embodiments, the thieno[2,3-d]pyrimidine derivative is administered at a dosage between 1 and 100 mg / kg. In some embodiments, the thieno[2,3-d]pyrimidine derivative is administered at a dosage between 1 and 50 mg / kg. In some embodiments, the thieno[2,3-d]pyrimidine derivative is administered at a dosage between 1 and 30 mg / kg. In some embodiments, the thieno[2,3-d]pyrimidine derivative is administered at a dosage between 1 and 20 mg / kg. In some embodiments, the thieno[2,3-d]pyrimidine derivative is administered at a dosage between 10 and 20 mg / kg. In some embodiments, the thieno[2,3-d]pyrimidine derivative is administered at a dosage of 17 mg / kg.

[0012] In some embodiments, the thieno[2,3-d]pyrimidine derivative is administered daily. In some embodiments, the thieno[2,3-d]pyrimidine derivative is administered weekly. In some embodiments, the thieno[2,3-d]pyrimidine derivative is administered for at least 26 weeks. In some embodiments, the thieno[2,3-d]pyrimidine derivative is provided as a pharmaceutical composition comprising a pharmaceutically acceptable carrier. In some embodiments, the method further comprises administering luteinizing hormone (LH) to a subject in need thereof.

[0013] In another aspect, the present disclosure provides a general formula I:

[0014]

Chemical formula

[0015] Or a pharmaceutically acceptable salt thereof (wherein R1 and R2, together with the nitrogen atom to which they are attached, optionally form a ring having 2 to 6 carbon atoms optionally containing one or more heteroatoms selected from N, O and / or S), a method for treating diet-induced obesity in a subject in need thereof, comprising the step of administering a thieno[2,3-d]pyrimidine derivative to the adipose tissue of the subject in need thereof. In some embodiments, the thieno[2,3-d]pyrimidine derivative is tert-butyl 5-amino-2-methylthio-4-(3-(2-(azetidin-1-yl)-acetamido)-phenyl)-thieno[2,3-d]pyrimidine-6-carboxamide; tert-butyl 5-amino-2-methylthio-4-(3-(2-(morpholin-4-yl)-acetamido)-phenyl)-thieno[2,3-d]pyrimidine-6-carboxamide; tert-butyl 5-amino-2-methylthio-4-(3-(2-(thiomorpholin-4-yl)-acetamido)-phenyl)-thieno[2,3-d]pyrimidine-6-carboxamide; tert-butyl 5-amino-2-methylthio-4-(3-(2-(piperidin-1-yl)-acetamido)-phenyl)-thieno[2,3-d]pyrimidine-6-carboxamide; tert-butyl 5-amino-2-methylthio-4-(3-(2-(pyrrolidin-1-yl)-acetamido)-phenyl)-thieno[2,3-d]pyrimidine-6-carboxamide or tert-butyl 5-amino-2-methylthio-4-(3-(2-(piperazin-1-yl)-acetamido)-phenyl)-thieno[2,3-d]pyrimidine-6-carboxamide, a compound selected from the group consisting of. In some embodiments, the thieno[2,3-d]pyrimidine derivative is ORG 43902.

[0016] In some embodiments, the thieno[2,3-d]pyrimidine derivative is administered at a dosage between 1 and 100 mg / kg. In some embodiments, the thieno[2,3-d]pyrimidine derivative is administered at a dosage between 1 and 50 mg / kg. In some embodiments, the thieno[2,3-d]pyrimidine derivative is administered at a dosage between 1 and 30 mg / kg. In some embodiments, the thieno[2,3-d]pyrimidine derivative is administered at a dosage between 1 and 20 mg / kg. In some embodiments, the thieno[2,3-d]pyrimidine derivative is administered at a dosage between 10 and 20 mg / kg. In some embodiments, the thieno[2,3-d]pyrimidine derivative is administered at a dosage of 17 mg / kg.

[0017] In some embodiments, the thieno[2,3-d]pyrimidine derivative is administered daily. In some embodiments, the thieno[2,3-d]pyrimidine derivative is administered weekly. In some embodiments, the thieno[2,3-d]pyrimidine derivative agonist is administered for at least 26 weeks. In some embodiments, the thieno[2,3-d]pyrimidine derivative is provided as a pharmaceutical composition comprising a pharmaceutically acceptable carrier. In some embodiments, the method further comprises administering luteinizing hormone (LH) to a subject in need thereof.

[0018] In another aspect, the present disclosure provides a method of treating obesity in a subject in need thereof, comprising administering to the subject in need thereof a combination of ORG 43553 and ORG 43902.

[0019] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although the methods and materials used in the present invention are described herein, other suitable methods and materials known in the art can also be used. The materials, methods, and examples are illustrative only and not intended to be limiting. All publications, patent applications, patents, sequences, database entries, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, this specification, including definitions, will control.

[0020] Other features and advantages of the present invention will become apparent from the following detailed description, drawings, and claims.

Brief Description of the Drawings

[0021]

Figure 1

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Figure 9

Figure 10

MODE FOR CARRYING OUT THE INVENTION

[0022] Luteinizing Hormone (LH) and Human Chorionic Gonadotropin (hCG) Receptors The luteinizing hormone / choriogonadotropin receptor (LHCGR) is a member of the G-protein coupled receptor (GPCR) subfamily characterized by the presence of a large N-terminal extracellular domain containing several leucine-rich repeats (LRRs). This glycoprotein hormone receptor family is named the LRR-containing GPCR (LGR) family. The human LHCGR protein is encoded by the LHCGR gene located at cytogenetic position 2p16.3, genomic coordinates (GRCh38): 2:48,686,774 - 48,755,724. The amino acid sequence of the human LHCGR precursor protein is provided below as SEQ ID NO: 1.

[0023]

Chemical Structure

[0024] In some embodiments, the methods disclosed herein include administering to a subject one or more LHCGR agonists. In some embodiments, the LHCGR agonist is a thieno[2,3-d]pyrimidine derivative. In some embodiments, the thieno[2,3-d]pyrimidine derivative has the general formula I:

[0025]

Chemical Structure

[0026] Or a compound according to a pharmaceutically acceptable salt thereof. In some embodiments, R1 and R2, together with the nitrogen atom to which they are attached, optionally contain one or more heteroatoms selected from N, O and / or S and form a ring having 2 to 6 carbon atoms. In some embodiments, the thieno[2,3-d]pyrimidine derivative is tert-butyl 5-amino-2-methylthio-4-(3-(2-(azetidin-1-yl)-acetamido)-phenyl)-thieno[2,3-d]pyrimidine-6-carboxamide; tert-butyl 5-amino-2-methylthio-4-(3-(2-(morpholin-4-yl)-acetamido)-phenyl)-thieno[2,3-d]pyrimidine-6-carboxamide; tert-butyl 5-amino-2-methylthio-4-(3-(2-(thiomorpholin-4-yl)-acetamido)-phenyl)-thieno[2,3-d]pyrimidine-6-carboxamide; tert-butyl 5-amino-2-methylthio-4-(3-(2-(piperidin-1-yl)-acetamido)-phenyl)-thieno[2,3-d]pyrimidine-6-carboxamide; tert-butyl 5-amino-2-methylthio-4-(3-(2-(pyrrolidin-1-yl)-acetamido)-phenyl)-thieno[2,3-d]pyrimidine-6-carboxamide or tert-butyl 5-amino-2-methylthio-4-(3-(2-(piperazin-1-yl)-acetamido)-phenyl)-thieno[2,3-d]pyrimidine-6-carboxamide, a compound selected from the group consisting of.

[0027] In some embodiments, the thieno[2,3-d]pyrimidine derivative is ORG 43533. ORG 43553 is an orally active and low molecular weight (LMW) LHCGR agonist. ORG 43553 exhibits agonist activity against the human LHCGR, and the EC 50 value is 3.7 nM. The structure of ORG 43553 is provided below:

[0028]

Chemical formula

[0029] Pharmaceutical composition In some embodiments, the compositions and methods described herein include the manufacture and use of pharmaceutical compositions and medicaments comprising one or more of the compounds disclosed herein. The pharmaceutical compositions themselves are also included.

[0030] In some embodiments, the compositions disclosed herein can include other compounds, drugs, or agents used in therapy. For example, in some instances, the pharmaceutical compositions disclosed herein can be combined with one or more (e.g., one, two, three, four, five, or less than ten) compounds.

[0031] In some embodiments, the pH of the compositions disclosed herein can be adjusted with pharmaceutically acceptable acids, bases, or buffers to enhance the stability of the compound or its delivery form.

[0032] Pharmaceutical compositions typically include a pharmaceutically acceptable carrier, adjuvant, or vehicle. As used herein, the term "pharmaceutically acceptable" is generally considered to be physiologically acceptable and refers to molecular entities and compositions that do not typically cause allergic or similar adverse reactions such as nausea, dizziness, etc. when administered to a human. A pharmaceutically acceptable carrier, adjuvant, or vehicle is a composition that can be administered to a patient together with the compounds of the present invention, does not destroy their pharmacological activity, and is non-toxic when administered in a dosage sufficient to deliver a therapeutic amount of the compound. Exemplary conventional non-toxic pharmaceutically acceptable carriers, adjuvants, and vehicles include saline, solvents, dispersion media, coating agents, antibacterial and antifungal agents, isotonic and absorption delaying agents, etc., which are compatible with pharmaceutical administration.

[0033] In particular, pharmaceutically acceptable carriers, adjuvants and vehicles that can be used in the pharmaceutical composition of the present invention include ion exchangers, alumina, aluminum stearate, lecithin, self-emulsifying drug delivery systems (SEDDS), for example d-α-tocopherol polyethylene glycol 1000 succinate, surfactants used in pharmaceutical dosage forms such as Tween or other similar polymer delivery matrices, serum proteins such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethyl cellulose, polyacrylate, wax, polyethylene-polyoxypropylene-block polymer, polyethylene glycol and lanolin, but are not limited thereto. Cyclodextrins such as α-, β- and γ-cyclodextrins can also be advantageously used to enhance the delivery of the compounds of the formulas described herein.

[0034] As used herein, a compound disclosed herein is defined to include its pharmaceutically acceptable derivatives or prodrugs. "Pharmaceutically acceptable derivative" means any pharmaceutically acceptable salt, solvate, or prodrug, such as a carbamate, ester, phosphate ester, salt of an ester, or other derivative of a compound or agent disclosed herein that can (directly or indirectly) provide a compound described herein, or its active metabolite or residue, upon administration to a recipient. Particularly preferred derivatives and prodrugs are those that increase the bioavailability of a compound disclosed herein when such compound is administered to a mammal (e.g., by making an orally administered compound more readily absorbed into the blood), or that enhance delivery of the parent compound to a biological compartment (e.g., the brain or lymphatic system) compared to the parent species. Preferred prodrugs include derivatives in which a group that enhances water solubility or active transport across the intestinal membrane is attached to the structure of the formula described herein. Such derivatives will be recognizable to one of ordinary skill in the art without undue experimentation. However, for the scope of teaching such derivatives, reference is made to the teachings of Burger's Medicinal Chemistry and Drug Discovery, 5th Edition, Volume 1: Principles and Practice, which is incorporated herein by reference.

[0035] Compounds disclosed herein include pure enantiomers, mixtures of enantiomers, pure diastereoisomers, mixtures of diastereoisomers, diastereoisomeric racemates, mixtures of diastereoisomeric racemates, and meso forms, as well as their pharmaceutically acceptable salts, solvate complexes, morphological forms, or deuterated derivatives.

[0036] In particular, pharmaceutically acceptable salts of the compounds disclosed herein include, for example, those derived from pharmaceutically acceptable inorganic and organic acids and bases. Examples of suitable acid salts include acetate, adipate, benzoate, benzenesulfonate, butyrate, citrate, digluconate, dodecyl sulfate, formate, fumarate, glycolate, hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide, hydroiodide, lactate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, palmoate, phosphate, picrate, pivalate, propionate, salicylate, succinate, sulfate, tartrate, tosylate, trifluoromethylsulfonate, and undecanoate. Salts derived from suitable bases include, for example, alkali metals (e.g., sodium), alkaline earth metals (e.g., magnesium), and ammonium salts. The present invention also contemplates the quaternization of any basic nitrogen-containing group of the compounds disclosed herein. Water-soluble or oil-soluble or dispersible products can be obtained by such quaternization.

[0037] In some embodiments, the pharmaceutical compositions disclosed herein can comprise an effective amount of one or more compounds. The terms "effective amount" and "therapeutically effective," as used herein, refer to the amount or concentration of one or more of the compounds or pharmaceutical compositions described herein that is effective within the context of its administration to cause the intended effect or physiological result over a period of time (including acute or chronic administration and periodic or continuous administration). In some embodiments, the pharmaceutical composition can further comprise one or more additional compounds, drugs, or agents that are used in treatment in an effective amount to cause the intended effect or physiological result.

[0038] In some embodiments, the pharmaceutical compositions disclosed herein can be formulated for sale within the United States, import into the United States, or export from the United States.

[0039] The present disclosure also encompasses the therapeutic combinations disclosed herein in the form of a kit or a packaged formulation. As used herein, a kit or a packaged formulation includes one or more dosages of the subject peptide and its salts in a container that holds the dosage, together with instructions for co - or sequential administration to a patient. For example, the package can contain the peptide together with a pharmaceutical carrier combined in the form of a powder for mixing with an aqueous solution and can be ingested by an affected subject. The package or kit includes appropriate instructions, which can include diagrams, records (e.g., audio, video, compact disk) and computer programs that provide instructions on the use of the combination therapy. The foregoing description of specific embodiments of the present disclosure has been presented for purposes of illustration and description. It is not intended to be exhaustive or to limit the disclosure to the precise forms disclosed, and it is obvious that many modifications and variations are possible in light of the above teachings.

[0040] Administration of Pharmaceutical Compositions The pharmaceutical compositions disclosed herein can be formulated or adapted for administration to a subject via any route, for example, any route approved by the Food and Drug Administration (FDA). Exemplary methods are described in the FDA Data Standards Manual (DSM). In particular, the pharmaceutical compositions can be formulated and administered for oral, parenteral, or transdermal delivery. As used herein, the term "parenteral" includes subcutaneous, intradermal, intravenous, intramuscular, intraperitoneal, intra - articular, intra - arterial, intrasynovial, intrasternal, intrasynovial, intralesional, and intracranial injection or infusion techniques.

[0041] For example, the pharmaceutical compositions disclosed herein can be administered, for example, topically, rectally, nasally (e.g., by inhalation spray or nebulizer), buccally, vaginally, subcutaneously (e.g., by injection or via an implant reservoir), or ophthalmically.

[0042] For example, the pharmaceutical composition of the present invention can be orally administered in any orally acceptable dosage form including, but not limited to, capsules, tablets, emulsions, and aqueous suspensions, dispersions, and solutions. In the case of tablets for oral use, commonly used carriers include lactose and corn starch. Lubricants such as magnesium stearate are also commonly added. When orally administered in capsule form, useful diluents include lactose and dried corn starch. When an aqueous suspension or emulsion is orally administered, the active ingredient may be suspended or dissolved in an oily phase, which may be combined with an emulsifying or suspending agent. Optionally, certain sweetening, flavoring, or coloring agents can be added.

[0043] For example, the pharmaceutical composition of the present invention can be administered in the form of suppositories for rectal administration. These compositions can be prepared by mixing the compounds of the present invention with suitable non-irritating excipients that are solid at room temperature but liquid at rectal temperature and thus melt within the rectum to release the active ingredient. Such materials include, but are not limited to, cocoa butter, beeswax, and polyethylene glycol.

[0044] For example, the pharmaceutical composition of the present invention can be administered by nasal aerosol or inhalation. Such compositions can be prepared according to techniques well known in the art of pharmaceutical formulations and can be prepared as solutions in physiological saline using benzyl alcohol or other suitable preservatives, absorption promoters to enhance bioavailability, fluorocarbons, or other solubilizing or dispersing agents well known in the art.

[0045] For example, the pharmaceutical composition of the present invention can be administered by injection (e.g., as a solution or powder). Such compositions can be formulated according to techniques known in the art using suitable dispersing or wetting agents (e.g., Tween 80, etc.) and suspending agents. The sterile injectable preparation can also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, for example, a solution in 1,3 - butanediol. Acceptable vehicles and solvents that can be used include mannitol, water, Ringer's solution, and isotonic sodium chloride solution. Mannitol, water, Ringer's solution, aqueous isotonic sodium chloride solution, and solvents are acceptable. Further, a sterile fixed oil is conventionally used as a solvent or suspending medium. For this purpose, any non-irritating fixed oil containing synthetic monoglycerides or diglycerides can be used. Fatty acids such as oleic acid and its glyceride derivatives are useful in the preparation of injectables, and natural pharmaceutically acceptable oils such as olive oil or castor oil, especially their polyoxyethylated versions, are also useful. These oil solutions or suspensions can also contain long-chain alcohol diluents or dispersing agents, or carboxymethyl cellulose or similar dispersing agents, which are commonly used in the formulation of pharmaceutically acceptable dosage forms such as emulsions and / or suspensions. Other commonly used surfactants such as Tweens, Spans, or other similar emulsifiers, or bioavailability enhancers, which are commonly used in the manufacture of pharmaceutically acceptable solid, liquid, or other dosage forms, can also be used for the purpose of formulation.

[0046] In some embodiments, the effective dosage of the pharmaceutical composition of the present invention can include, without limitation, for example, about 0.00001, 0.0001, 0.001, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.15, 0.2, 0.25, 0.3, 0.35, 0.4, 0.45, 0.5, 0.55, 0.6, 0.65, 0.7, 0.75, 0.8, 0.85, 0.9, 0.95, 1, 1.25, 1.5, 1.75, 2, 2.5, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30, 40, 50, 60, 70, 80, 90, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000, 2500, 5000, or 10000 mg / kg / day, or it can follow the requirements of a specific pharmaceutical composition.

[0047] When the pharmaceutical composition disclosed herein comprises a combination of a compound of the formulations described herein and one or more additional compounds (e.g., one or more additional compounds, drugs, or agents used in the treatment of obesity or any other obesity-related condition or disease, including conditions or diseases known to be associated with or caused by obesity), both the compound and the additional compound should be present at dosage levels between about 1% and 100%, more preferably between about 5% and 95% of the dosage normally administered in a monotherapy regimen. The additional agent can be administered as part of a multiple dosing regimen separately from the compound of the present invention. Alternatively, these agents can be mixed with the compound of the present invention to form a single composition and be part of a single dosage form.

[0048] In some embodiments, the pharmaceutical composition disclosed herein can be included in a container, pack, or dispenser together with instructions for administration.

[0049] Combination therapy In various embodiments, the compounds or compositions described herein can be administered to a subject in combination with one or more other therapies (e.g., biological or antibody therapies). In some embodiments, the pharmaceutical compositions described herein can be administered to a subject in combination with one or more therapies. The one or more other therapies may have the same composition as the compounds described herein or a different composition.

[0050] In certain embodiments, the therapeutic agent is administered at intervals of less than 5 minutes, less than 30 minutes, 1 hour, about 1 hour, about 1 to about 2 hours, about 2 to about 3 hours, about 3 to about 4 hours, about 4 to about 5 hours, about 5 to about 6 hours, about 6 to about 7 hours, about 7 to about 8 hours, about 8 to about 9 hours, about 9 to about 10 hours, about 10 to about 11 hours, about 11 to about 12 hours, about 12 to 18 hours, 18 to 24 hours, 24 to 36 hours, 36 to 48 hours, 48 to 52 hours, 52 to 60 hours, 60 to 72 hours, 72 to 84 hours, 84 to 96 hours, or 96 to 120 hours. In some embodiments, two or more therapeutic agents are administered simultaneously. The two or more therapeutic agents can be administered in the same composition or in different compositions. Further, the two or more therapeutic agents can be administered by the same route of administration or by different routes of administration.

[0051] Method of treatment The methods disclosed herein contemplate the administration of an effective amount of a compound or composition to achieve the desired or described effect. Typically, the compounds or compositions of the invention are administered about 1 to about 6 times per day, or alternately, or additionally, as a continuous infusion. Such administration can be used as chronic therapy or acute therapy. The amount of active ingredient that can be combined with a carrier material to produce a single dosage form will vary depending on the host being treated and the particular mode of administration. Typical formulations contain from about 5% to about 95% active compound (w / w). Alternatively, such formulations can contain from about 20% to about 80% active compound.

[0052] In some embodiments, the disclosure provides a method for using a composition comprising a compound, the method comprising the pharmaceutical composition (hereinafter referred to as "X") disclosed herein in the following methods:

[0053] Substance X for use as a medicament in the treatment of one or more of the diseases or conditions disclosed herein. Use of substance X for the manufacture of a medicament for the treatment of Y; and substance X for use in the treatment of Y.

[0054] In some embodiments, the disclosed method comprises administering a therapeutically effective amount of one or more of the compounds or compositions described herein to a subject (e.g., a mammalian subject, e.g., a human subject) determined to be in need of or in need of such treatment. In some embodiments, the disclosed method comprises the steps of selecting a subject, administering to the subject an effective amount of one or more of the compounds or compositions described herein, and optionally repeating the administration as needed for the prevention or treatment of obesity or obesity-related diseases.

[0055] In some embodiments, subject selection can include obtaining a sample from a subject (e.g., a candidate subject) and testing the sample to indicate that the subject is suitable for selection. In some embodiments, a subject can be confirmed or identified as having or having had a condition or disease, e.g., by a medical professional. In some embodiments, suitable subjects include, for example, subjects who had or have had a condition or disease, but the disease or its aspect has disappeared and the symptoms of the disease have been alleviated (e.g., compared to other subjects having the same condition or disease (e.g., the majority of the subjects)), or, for example, subjects who have survived for a long time with that condition or disease in an asymptomatic state (e.g., compared to other subjects having the same condition or disease (e.g., the majority of the subjects)). In some embodiments, the presentation of a positive immune response to a condition or disease can be made from patient records, family history, or detection of signs of a positive immune response. In some embodiments, multiple stakeholders can be involved in subject selection. For example, a first stakeholder can obtain a sample from a candidate subject and a second stakeholder can examine the sample. In some embodiments, a subject can be selected or referred by a practicing physician (e.g., a general practitioner). In some embodiments, subject selection can include obtaining a sample from a selected subject and storing the sample or using the sample in the methods disclosed herein. The sample can include, for example, cells or a population of cells.

[0056] In some embodiments, treatment methods can include single, multiple, and repeated administrations of one or more compounds disclosed herein that are required for the prevention or treatment of a disease or condition that a subject is suffering from. In some embodiments, treatment methods can include assessing the level of the disease in the subject before, during, or after treatment. In some embodiments, treatment can be continued until a decrease in the level of the disease in the subject is detected.

[0057] As used herein, the term "subject" refers to any animal. In some examples, the subject is a mammal. In some examples, as used herein, the term "subject" refers to a human (e.g., male, female, or child).

[0058] As used herein, the terms "administer", "administering", or "administration" refer to implanting, ingesting, injecting, inhaling, or otherwise absorbing a compound or composition, regardless of form. For example, the methods disclosed herein include administration of an effective amount of a compound or composition to achieve the desired or described effect.

[0059] As used herein, the terms "treat", "treating", or "treatment" refer to partially or completely alleviating, suppressing, improving, or eliminating a disease or condition with which the subject is afflicted. This means any method by which one or more of the symptoms of a disease or disorder are improved or otherwise beneficially changed. As used herein, improvement of the symptoms of a particular disorder refers to any alleviation, whether permanent or temporary, persistent or transient, that may result from or be related to treatment with the compositions and methods of the present invention.

[0060] As used herein, the terms "prevent", "preventing", and "prevention" are intended to refer to a decrease in the occurrence of a disease in a subject, or a decrease in the risk of acquiring a disease or its associated symptoms. Prevention may be complete, e.g., prevention in which no disease or diseased cells are present in the subject. Prevention may also be partial, such that the occurrence of a disease or diseased cells in the subject is less, or later, or the onset is more mild, than would have occurred in the absence of the present invention.

[0061] As used herein, the term "prevent a disease" in a subject means, for example, preventing the onset of one or more diseases in the subject before the symptoms of the one or more diseases in the subject become detectable, for example, by occurrence or by detection by the patient or the patient's attending physician. Preferably, the disease does not develop at all, i.e., the symptoms of the disease cannot be detected. However, it can also mean delaying or retarding the onset of one or more symptoms of the disease. Alternatively, or additionally, it can mean reducing the severity of one or more subsequently developing symptoms.

[0062] The specific dosage and treatment regimen for a particular patient depend on a variety of factors, including the activity of the particular compound used, age, weight, general health, sex, diet, time of administration, rate of excretion, combination of drugs, severity and course of the disease, condition or symptom, the patient's susceptibility to the disease, condition or symptom, and the judgment of the physician in charge of the treatment, etc.

[0063] An effective amount can be administered in one or more administrations, applications or dosings. The therapeutically effective amount (i.e., the effective dosage) of a therapeutic compound depends on the therapeutic compound selected. Further, the treatment of a subject with a therapeutically effective amount of a compound or composition described herein can include a single treatment or a series of treatments. For example, the effective amount can be administered at least once. The composition can be administered once every other day; from once or multiple times a day to once or multiple times a week; including once a day. One of ordinary skill in the art will understand that specific factors, including but not limited to the severity of the disease or disorder, previous treatment, the general health or age of the subject, and the presence of other diseases, can affect the dosage and timing required to effectively treat the subject.

[0064] After administration, the subject can be evaluated to detect, assess, and determine the level of the disease. In some examples, treatment can be continued until a change (e.g., a decrease) in the disease level of the subject is detected. Upon improvement of the patient's condition (e.g., a change (e.g., a decrease) in the level of the disease in the subject), a maintenance dose of the compounds or compositions disclosed herein can be administered as needed. Thereafter, the dosage or dosing frequency, or both, can be decreased, for example as a function of symptoms, to a level at which the improved condition is maintained. However, the patient may require long-term intermittent treatment if the symptoms of the disease recur.

[0065] The publications disclosed herein are provided only for disclosures prior to the filing date of the present disclosure. Nothing in this specification shall be construed as an admission that the present disclosure has no right to antedate such publications. Further, the provided publication dates may be different from the actual publication dates, which may need to be independently verified.

[0066] Each application and patent cited in the text, and each document or reference cited in each application and patent, patent or non-patent literature (including each issued patent under examination; hereinafter "application cited references"), and each PCT application and foreign application or patent corresponding to any of these applications and patents and / or claiming priority from any of these applications and patents, and each document cited or referenced in each application cited reference are hereby expressly incorporated by reference in their entirety into this specification. More generally, whether in the reference list preceding the claims or in the text itself, where a document or reference is cited in the text, each of these documents or references ("this specification cited references") and each document or reference cited in each of these specification cited references (including manufacturer's specifications, instructions, etc.) are hereby expressly incorporated by reference into this specification.

[0067] The following non-limiting examples are useful for further illustrating the present disclosure.

Example

[0068] The present invention will be further described in the following examples, which do not limit the scope of the invention described in the claims.

[0069] Method In the following examples, the following materials and methods were used.

[0070] qPCR: Total RNA was isolated and reverse transcribed using TRIzol reagent (Invitrogen, 15596018) (Invitrogen, 18064014). qPCR was performed using SYBR Green Master Mix (Applied Biosystems 4472918).

[0071] Sanger sequencing: RNA from mouse genital adipose tissue was reverse transcribed (Invitrogen 18080093). Fragments of Lhcgr were amplified by PCR and sequenced.

[0072] RNASCOPE (trademark): Paraffin sections were processed and stained using RNAscope (trademark) Probe-Mm-Lhcgr (ACD 408171) and RNAscope (trademark) 2.5HD Reagent Kit-Brown (ACD 322300). IHC: Paraffin sections were stained using rabbit LHCGR polyclonal antibody (Invitrogen, PA5-79598) and anti-rabbit HRP-IgG (Leica RE7161).

[0073] Biodistribution: Male and female C57BL / 6 mice (5 mice / group) were 89 intravenously injected with Zr-LH, and after 2 hours, each tissue was dissected and gamma counted.

[0074] hCG binding: Recombinant hCG was conjugated using Alexa Fluor 488 Microscale Protein Labelling Kit (Invitrogen A30006) and intraperitoneally injected (0.45 μg).

[0075] Cell culture: Differentiated 3T3-L1 preadipocytes (Zen-Bio) were treated with hCG or LH for 20 minutes.

[0076] siRNA knockdown: Lentiviral vectors expressing Lhcgr siRNA (ABM 2649609) or siControl (ABM LVP015-G) were used according to the manufacturer's recommendations.

[0077] Western blotting: Proteins were blotted using commercially available antibodies (Cell Signaling, Phospho-ERK1 / 2, 9101S; ERK1 / 2, 4695S, 1:5000).

[0078] Statistics: Two-sided Student's t-test or two-way ANOVA was used (*p < 0.05; **p < 0.01).

[0079] (Example 1) Expression, binding, and activation of Lhcgr in mouse adipocytes qPCR was used to measure the relative expression in various male and female mouse fat depots, and the results were plotted in Figure 1A. As shown in Figure 1B, the sequence of the extracellular domain of Lhcgr in mouse gonadal white adipose tissue (gWAT) was identical to that in the ovarian receptor. As shown in Figure 1C, RNAscope (trademark) in situ hybridization and immunohistochemistry (IHC) were used to visualize the expression of LHCGR in various mouse fat depots and gonads of male and female C57BL / 6 mice. Lhcgr - / - No staining was detected in mice, indicating the specificity of the LHCGR probe and antibody. As shown in Figure 1D, significant concentrations of 89 Zr-LH were detected in multiple organs including mesenteric and gonadal WAT. As shown in Figure 1E, intraperitoneally injected AlexaFluor-488-labeled hCG localized to mouse iWAT and gWAT. This hCG binding further confirmed the presence of LHCGR in adipose tissue.

[0080] (Example 2) LHCGR is expressed and fully functional in adipocytes As shown in Figure 2A, the relative Lhcgr expression in mouse iWAT and 3T3-L1 adipocytes measured by qPCR indicates that Lhcgr is expressed in mouse iWAT and 3T3-L1 adipocytes. As shown in Figure 2B, RNAscope™ in situ hybridization and immunohistochemistry (IHC) show LHCGR staining in differentiated 3T3-L1 murine adipocytes. As shown in Figure 2C, the specificity of the antibody was confirmed by siRNA-lentiviral knockdown of Lhcgr (Cy-3, red) in 3T3-L1 adipocytes. The GFP reporter (green) showed a high efficiency of lentiviral infection. As shown in Figure 2D, the binding of AlexaFluor-488-labeled hCG to differentiated 3T3-L1 cells was specific, as this binding could be competitively displaced by pretreatment with unlabeled hCG. As shown in Figure 2E, LH and hCG induced ERK1 / 2 phosphorylation in a dose-dependent manner in differentiated 3T3L1 adipocytes, suggesting that adipose LHCGR is fully functional. The ERK1 / 2 phosphorylation induced by hCG disappeared upon LHCGR siRNA knockdown, further suggesting that ERK1 / 2 phosphorylation by LH acts via LHCGR in adipocytes. RNAseq was performed to measure the log2(fold change) of Lep, Cebpb, Cebpa, Ucp2, and Ppargc1b adipose genes after treatment of differentiated 3T3-L1 adipocytes with 1 nM LH or hCG for 12 hours. As shown in Figure 2F, several adipose genes including leptin (Lep) were downregulated. As shown in Figure 2G, treatment with hCG decreased the accumulation of oil droplets (Oil Red O staining) in differentiated 3T3-L1 adipocytes.

[0081] (Example 3) Lhcgr deficiency causes weight gain in female mice on a normal diet and male mice on a high-fat diet (HFD) Male and female Lhcgr + / + , Lhcgr + / -and Lhcgr - / - Mice were fed a normal diet or a high-fat diet for 40 weeks. As shown in Figure 3A, female Lhcgr mice fed a normal diet + / - and Lhcgr - / - Mice, Lhcgr + / + The mice showed higher fat mass compared to their littermates, indicating the anti-adipose effect of LH. - / - Mouse Lhcgr + / + They also showed higher total weight compared to Lhcgr mice. As shown in Figure 3D, H&E staining revealed that Lhcgr mice - / - and Lhcgr + / - In mouse iWAT and gWAT, Lhcgr + / + As shown in Figures 3B, 3E, and 3F, there were no differences in lean body mass, glucose tolerance, or serum estrogen levels among the three genotypes. As shown in Figure 3G, on a high-fat diet, male Lhcgr mice showed significantly larger adipocytes than their littermates. + / - and Lhcgr - / - Mice, Lhcgr + / + As shown in Figure 3I, there was no difference in total body weight among the three genotypes. As shown in Figure 3J, male Lhcgr mice fed a high-fat diet showed a higher fat mass than male Lhcgr mice. - / - Mice and Lhcgr + / - In mice, H&E staining showed that adipocytes were larger in iWAT and gWAT. As shown in Figure 3H, male Lhcgr mice on a HFD had larger adipocytes. - / - Mice, Lhcgr + / + and Lhcgr + / - As shown in Figure 3K, glucose tolerance was similar among the different genotypes.

[0082] Example 4 LH and hCG reduce HFD-induced weight gain in C57BL / 6 mice Fourteen-week-old C57BL / 6 mice (20 mice / group) were treated with vehicle (200 μL of saline), LH (250 ng / mouse), or hCG (250 ng / mouse) by intraperitoneal injection and fed a high-fat diet (TestDiet 0056833) for 6 weeks. As shown in FIGS. 4A and 4E, when high-dose LH was injected twice a week into 14-week-old C57BL / 6 male mice fed a high-fat diet, the body fat mass was significantly decreased. As shown in FIG. 4B, the same treatment increased the fat-free weight, and as shown in FIG. 4C, the same treatment decreased the total weight. As shown in FIG. 4D, the food intake was comparable in all groups. As shown in FIG. 4A, the hCG-treated mice showed a lower fat mass, and as shown in FIG. 4E, the hCG-treated mice showed a lower iWAT mass. As shown in FIG. 4F, there was no difference in glucose tolerance among the three treatment groups. As shown in FIG. 4G, the mice treated with LH or hCG showed an increase in testosterone. As shown in FIG. 4H, the mice treated with hCG and LH had lower leptin levels.

[0083] (Example 5) The anti-obesity effect of LH / hCG is independent of testosterone To evaluate the anti-obesity effect of LH / hCG on testosterone levels, the androgen receptor antagonist flutamide was used to block the increase in testosterone after LH / hCG treatment in mice. Fourteen-week-old C57BL / 6 mice (15 mice / group) were given the following intraperitoneal injections - Group I: vehicle and placebo (placebo 60-day release pellet); Group II: vehicle and flutamide (7.5 mg 60-day release pellet); Group III: hCG (250 ng / mouse) and placebo; Group IV: hCG and flutamide. Mice were fed a high-fat diet for 6 weeks. As shown in FIGS. 5A - 5C, hCG decreased diet-induced fat mass (A) in the presence of flutamide, but did not decrease lean mass (B) or total mass (C), indicating that testosterone does not play a role in the action of hCG to reduce body fat. As shown in FIG. 5E, mice treated with hCG had lower iWAT and gWAT mass in both the flutamide group and the placebo group. As shown in FIGS. 5D and 5F, food intake (D) and glucose tolerance (F) were similar in all groups. As shown in FIG. 5G, mice treated with hCG had lower leptin levels in both the flutamide group and the placebo group.

[0084] (Example 6) The LHCGR agonist ORG 43553 reduces body fat To evaluate the effect of treatment with the LHCGR agonist ORG 43553 on body fat in mice, fourteen-week-old C57BL / 6 mice (25 mice / group) were treated with vehicle or ORG 43553 (17 mg / kg) by subcutaneous injection and fed a high-fat diet for 9 weeks. As shown in FIGS. 6A - 6C, ORG 43553 decreased diet-induced fat mass (A) and total mass (B) in mice, but did not decrease lean mass (C). As shown in FIG. 6E, mice treated with ORG 43553 had lower iWAT, gWAT, and mWAT mass. Food intake (FIG. 6D), glucose tolerance (FIG. 6F), testosterone (FIG. 6G), and leptin levels (FIG. 6H) were similar in all groups.

[0085] (Example 7) LHCGR Expression in Adipose Tissue To characterize Lhcgr expression in adipose tissue, qPCR, RNAscope™ in situ hybridization and immunohistochemistry (IHC) of Lhcgr transcripts were performed in mouse adipocytes. The relative expression of Lhcgr transcripts in white adipose tissue (WAT) accumulation of male and female C57BL / 6 mice measured by qPCR is shown in Fig. 7A. Expression in gonadal WAT (gWAT) of female mice was close to that of the ovary itself (gonad). As shown in Fig. 7B, Lhcgr transcripts (dark dots) were confirmed by RNAscope™ in situ hybridization in both male and female wild-type mice in the gonad, gWAT and subcutaneous WAT (sWAT), but Lhcgr - / - was not confirmed in mice. As shown in Fig. 7C, expression was verified by Sanger sequencing of the LHCGR ectodomain, which was identical to the ovarian receptor. As shown in Fig. 7D, LHCGR expression was shown immunohistochemically in gWAT and sWAT.

[0086] Binding of labeled hCG in vivo was tested. As shown in Fig. 7E, intraperitoneal AlexaFluor-488 labeled hCG bound to the gonad and subcutaneous fat pads of wild-type mice, but not to LHCGR-deficient mice. Complementarily, 89 Zr-LH was injected intravenously, sacrificed 2 hours later, perfused with PBS, and then organ collection and gamma counting were performed. As shown in Fig. 7F, radioactivity was detected in mesenteric WAT (mWAT), inguinal WAT (iWAT) and gWAT among other organs including the gonad. Finally, as shown in Fig. 7G, LH, hCG and ORG 43553 rapidly induced ERK1 / 2 phosphorylation in a concentration-dependent manner in differentiated 3T3.L1 cells, but no signal was seen in siLhcgr cells as expected. These results indicated that adipose LHCGR is functional. In summary, adipose tissue LHCGR was shown to be a target of LH action.

[0087] (Example 8) LH Induces Weight Loss As shown in Fig. 8A, female Lhcgr mice fed a normal diet + / - showed an increase in fat without a decrease in serum estrogen when measured by EchoMRI (trademark)-100. Similarly, as shown in Fig. 8B, male Lhcgr + / - mice became obese on a high-fat diet (TestDiet0056833), but importantly, serum testosterone did not decrease. This gene dosage effect of Lhcgr on adiposity suggests a dominant physiological role of LH on body composition.

[0088] As shown in Fig. 8C, in a gain-of-function experiment, 14-week-old male C57BL / 6 mice (N = 20 / group) were fed a high-fat diet and injected intraperitoneally with LH (250 ng), hCG (250 ng) or vehicle (200 μL saline) twice a week for 6 weeks. Both LH and hCG significantly decreased the spontaneous increase in body fat when measured by EchoMRI (trademark)-100, but induced an increase in serum testosterone. As shown in Fig. 8D, the inability of LH (250 ng) to suppress the spontaneous increase in fat in Lhcgr - / - mice demonstrated its action via LHCGR - however, in this experiment, the effect of LH on adipose tissue LHCGR could not be isolated from its effect on Leydig cells producing testosterone. To address the confounding effect of high testosterone on body fat, the experiment was repeated in the presence of flutamide (7.5 mg, low-release pellet for 60 days). As shown in Fig. 8E, the lipolytic phenotype induced by hCG persisted despite androgen receptor blockade by flutamide, indicating that the anti-lipolytic action of LH and hCG is largely independent of testosterone.

[0089] Next, the LHCGR agonist ORG 43553 was injected into male C57BL / 6 mice (N = 24 / group) for 9 weeks (17 mg / kg, subcutaneous, twice a week). As shown in Fig. 8F, a significant decrease in fat mass was observed, with a decrease in iWAT, gWAT, and renal WAT (rWAT). Since serum testosterone did not change, it was shown that the anti-fat effect of ORG 43553 is not testosterone-dependent. As shown in Fig. 8G, the distribution of adipocytes in iWAT showed that in mice treated with ORG 43553, there were more adipocytes with an area below 4000 μm 2 and fewer adipocytes with an area exceeding 4000 μm 2 . As shown in Fig. 8H, when ORG 43553 was injected into a replicate cohort, fat mass and total weight significantly decreased along with a decrease in iWAT, gWAT, and mWAT, without dependence on testosterone. These data indicate that activation of LHCGR induces a lipolytic phenotype. Furthermore, male and female homozygous Lhcgr - / - mice showed more obvious obesity (data not shown), but this effect is likely to be confounded by concomitant hypogonadism, especially in older age.

[0090] (Example 9) LHCGR agonism inhibits adipocyte differentiation and induces thermogenesis To test the mechanism of LHCGR activation against body fat, 3T3-L1 preadipocytes and fat organoids were used. Oil Red O staining can be used to label lipid-containing vacuoles and quantify the lipid content of cells. As shown in Figure 9A, when 3T3-L1 adipocytes were treated with hCG (1 nM) or ORG 43553 (6 nM) for 12 hours, it was shown that the accumulation of oil droplets (Oil Red O staining) decreased and the lipid content decreased. The organoids were derived from 3T3-L1 cells cultured in Nunclon Sphera dishes for 8 days and exposed to LH, hCG or ORG 43553 for 3 days. After further differentiation for 7 days, paraffin sections were stained with H&E, and LHCGR expression was confirmed by immunostaining and RNAscope (trademark). The thickness of the "differentiated cell layer" was measured with QuPath-0.2.3. As shown in the plot of Figure 9B, the organoids treated with LH, hCG or ORG 43553 showed a significant decrease in the differentiated layer compared to the vehicle-treated organoids.

[0091] Next, 3T3-L1 adipocytes were treated with ORG 43553, and in addition to reducing adipocyte differentiation, it was determined whether LHCGR agonism also enhanced thermogenesis. Thus, after pre-treating 3T3-L1 adipocytes with ORG 43553 for 1 hour, the oxygen consumption rate (OCR) was measured using a Seahorse Xf96 Analyzer. As shown in Figure 9C, oligomycin (Oligo), FCCP, and rotenone / antimycin-A were added at the indicated times. ORG 43553 increased OCR at baseline and during oligomycin exposure, which indicates mitochondrial proton leak (a surrogate for thermogenesis). To confirm the increase in energy consumption in vivo, mice fed a high-fat diet (N = 6 / group) were acclimated to metabolic cages for 1 day, then injected subcutaneously with ORG 43553 (17 mg / kg, blue line) or vehicle, and monitored for 48 hours. As shown in Figure 9D, oxygen consumption volume (VO2) and energy expenditure (EE) increased acutely in ORG 43553-treated mice, and there was no change in locomotor activity (distance). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. These results indicate that the anti-obesity effect of LHCGR agonism results from a decrease in adipocyte differentiation and an increase in thermogenesis.

[0092] (Example 10) Structure of ORG 43553 and Binding to LHCGR Figure 10A shows the chemical structure of ORG 43553 [5-amino-2-methylsulfanyl-4-[3-(2-morpholin-4-yl-acetylamino)-phenyl]-thieno[2,3-d]pyrimidine-6-carboxylic acid tert-butylamide]. The binding of ORG 43553 to mouse LHCGR was computationally modeled. Unlike LH, which binds to the ectodomain, ORG 43553 binds to the transmembrane domain (TMD) (Figure 10B). Figure 10C shows a side view (left) and a top view (right) of ORG 43553 complexed with LHCGR by 280 nanosecond molecular dynamics.

[0093] Other Embodiments The present invention, while described in conjunction with its detailed description, is to be understood that the foregoing description is for the purpose of illustrating the scope of the present invention as defined by the appended claims and is not intended to limit the scope of the present invention. Other aspects, advantages, and modifications are within the scope of the following claims.

Claims

1. A composition for treating obesity in a person requiring treatment for obesity, wherein the composition comprises general formula I: 【Chemistry 1】 A composition comprising a thieno[2,3-d]pyrimidine derivative relating to a pharmaceutically acceptable salt thereof (wherein Rl and R2, together with the nitrogen atom to which they are bonded, form a ring having 2 to 6 carbon atoms, optionally containing one or more heteroatoms selected from N, O and / or S).

2. Thieno[2,3-d]pyrimidine derivatives include tert-butyl5-amino-2-methylthio-4-(3-(2-(azetidine-1-yl)-acetamide)-phenyl)-thieno[2,3-d]pyrimidine-6-carboxamide; tert-butyl5-amino-2-methylthio-4-(3-(2-(morpholine-4-yl)-acetamide)-phenyl)-thieno[2,3-d]pyrimidine-6-carboxamide; tert-butyl5-amino-2-methylthio-4-(3-(2-(thiomorpholine-4-yl)-acetamide)-phenyl)-thieno[2,3-d]pyrimidine-6-carboxamide; tert The composition according to claim 1, wherein the compound is selected from the group consisting of -butyl 5-amino-2-methylthio-4-(3-(2-(piperidine-1-yl)-acetamide)-phenyl)-thieno[2,3-d]pyrimidine-6-carboxamide; tert-butyl 5-amino-2-methylthio-4-(3-(2-(pyrrolidine-1-yl)-acetamide)-phenyl)-thieno[2,3-d]pyrimidine-6-carboxamide or tert-butyl 5-amino-2-methylthio-4-(3-(2-(piperazine-1-yl)-acetamide)-phenyl)-thieno[2,3-d]pyrimidine-6-carboxamide.

3. The composition according to claim 1, wherein the thieno[2,3-d]pyrimidine derivative is ORG 43533.

4. The composition according to claim 1, wherein the thieno[2,3-d]pyrimidine derivative is administered in a dose between 1 and 100 mg / kg.

5. The composition according to claim 4, wherein the thieno[2,3-d]pyrimidine derivative is administered in a dose between 1 and 50 mg / kg.

6. The composition according to claim 5, wherein the thieno[2,3-d]pyrimidine derivative is administered in a dose between 1 and 30 mg / kg.

7. The composition according to claim 6, wherein the thieno[2,3-d]pyrimidine derivative is administered in a dose between 1 and 20 mg / kg.

8. The composition according to claim 7, wherein the thieno[2,3-d]pyrimidine derivative is administered in a dose between 10 and 20 mg / kg.

9. The composition according to claim 8, wherein the thieno[2,3-d]pyrimidine derivative is administered at a dose of 17 mg / kg.

10. The composition according to claim 1, wherein a thieno[2,3-d]pyrimidine derivative is administered daily.

11. The composition according to claim 1, wherein a thieno[2,3-d]pyrimidine derivative is administered weekly.

12. The composition according to claim 1, wherein the thieno[2,3-d]pyrimidine derivative is administered for at least 26 weeks.

13. The composition according to claim 1, wherein a thieno[2,3-d]pyrimidine derivative is provided as a pharmaceutical composition comprising a pharmaceutically acceptable carrier.

14. The composition according to any one of claims 1 to 13, further comprising luteinizing hormone (LH).

15. A composition for treating diet-induced obesity in a person requiring treatment for diet-induced obesity, wherein the composition comprises general formula I: 【Chemistry 2】 A composition comprising a thieno[2,3-d]pyrimidine derivative relating to a pharmaceutically acceptable salt thereof (wherein Rl and R2, together with the nitrogen atom to which they are bonded, form a ring having 2 to 6 carbon atoms, optionally containing one or more heteroatoms selected from N, O and / or S).

16. Thieno[2,3-d]pyrimidine derivatives include tert-butyl5-amino-2-methylthio-4-(3-(2-(azetidine-1-yl)-acetamide)-phenyl)-thieno[2,3-d]pyrimidine-6-carboxamide; tert-butyl5-amino-2-methylthio-4-(3-(2-(morpholine-4-yl)-acetamide)-phenyl)-thieno[2,3-d]pyrimidine-6-carboxamide; tert-butyl5-amino-2-methylthio-4-(3-(2-(thiomorpholine-4-yl)-acetamide)-phenyl)-thieno[2,3-d]pyrimidine-6-carboxamide; tert- The composition according to claim 15, wherein the compound is selected from the group consisting of butyl 5-amino-2-methylthio-4-(3-(2-(piperidine-1-yl)-acetamide)-phenyl)-thieno[2,3-d]pyrimidine-6-carboxamide; tert-butyl 5-amino-2-methylthio-4-(3-(2-(pyrrolidine-1-yl)-acetamide)-phenyl)-thieno[2,3-d]pyrimidine-6-carboxamide or tert-butyl 5-amino-2-methylthio-4-(3-(2-(piperazine-1-yl)-acetamide)-phenyl)-thieno[2,3-d]pyrimidine-6-carboxamide.

17. The composition according to claim 15, wherein the thieno[2,3-d]pyrimidine derivative is ORG 43902.

18. The composition according to claim 15, wherein the thieno[2,3-d]pyrimidine derivative is administered in a dose between 1 and 100 mg / kg.

19. The composition according to claim 18, wherein the thieno[2,3-d]pyrimidine derivative is administered in a dose between 1 and 50 mg / kg.

20. The composition according to claim 19, wherein the thieno[2,3-d]pyrimidine derivative is administered in a dose between 1 and 30 mg / kg.

21. The composition according to claim 20, wherein the thieno[2,3-d]pyrimidine derivative is administered in a dose between 1 and 20 mg / kg.

22. The composition according to claim 21, wherein the thieno[2,3-d]pyrimidine derivative is administered in a dose between 10 and 20 mg / kg.

23. The composition according to claim 22, wherein the thieno[2,3-d]pyrimidine derivative is administered at a dose of 17 mg / kg.

24. The composition according to claim 15, wherein a thieno[2,3-d]pyrimidine derivative is administered daily.

25. The composition according to claim 15, wherein a thieno[2,3-d]pyrimidine derivative is administered weekly.

26. The composition according to claim 15, wherein a thieno[2,3-d]pyrimidine derivative agonist is administered for at least 26 weeks.

27. The composition according to claim 15, wherein the thieno[2,3-d]pyrimidine derivative is provided as a pharmaceutical composition comprising a pharmaceutically acceptable carrier.

28. The composition according to any one of claims 15 to 27, further comprising luteinizing hormone (LH).

29. A composition for treating obesity in subjects requiring treatment for obesity, comprising a combination of ORG 43553 and ORG 43902.

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