Ursolic acid morpholine salt and ursolic acid diethanolamine salt

Abstract

The present application provides ursolic acid diethanolamine salt and ursolic acid morpholine salt. It also provides a composition comprising the salt, and a method of using the salt.

Description

[0001] Cross - reference to related applications

[0002] This application claims the benefit of priority of U.S. Provisional Application No. 62 / 558,004, filed on September 13, 2017, and U.S. Provisional Application No. 62 / 649,938, filed on March 29, 2018. The entire content of each priority application is hereby incorporated herein by reference. Technical field

[0003] This application provides novel ursolic acid diethanolamine salts and ursolic acid morpholine salts, which exhibit excellent efficacy and properties. Compositions containing these salts and methods of using these salts are also provided. Background art

[0004] Ursolic acid is a pentacyclic triterpenoid acid. WO 2011 / 146768 and WO 2012 / 170546 discuss a series of biological actions of ursolic acid. At the molecular level, ursolic acid inhibits the STAT3 activation pathway, reduces the expression of matrix metalloproteinase - 9 through the glucocorticoid receptor, inhibits protein tyrosine phosphatase, acts as an insulin mimetic, activates PPARα, inhibits the NF - kB transcription factor, translocates hormone - sensitive lipase to stimulate lipolysis and inhibits the hepatic polyol pathway, among other actions. Summary of the invention

[0005] Briefly, this application provides novel ursolic acid diethanolamine salts and ursolic acid morpholine salts, as well as related compositions and methods. These salts exhibit excellent properties compared to ursolic acid itself and other ursolate salts, including effects on skeletal muscle.

[0006] Some embodiments of this application disclose ursolate salts, compositions containing the salt, and methods of using the salt, all of which have several characteristics, none of which alone is responsible for their desirable properties. Without limiting the scope of the ursolate salts, compositions containing the salt, and methods of using the salt as defined in the appended claims, their more prominent features will now be briefly discussed. After considering this discussion, and particularly after reading the section of this specification entitled "Detailed Description", one will understand that the features of the various embodiments disclosed herein provide many advantages over the prior art. For example, compared to, for example, natural ursolic acid and other ursolate salts, the embodiments of this application provide unexpectedly superior performance, including, for example, having the efficacy of stimulating skeletal muscle hypertrophy, increasing lean muscle mass, reducing fat mass, increasing skeletal muscle fiber size, reducing adipocyte size, reducing obesity and / or blood glucose, and improving glucose tolerance.

[0007] In a first aspect, this application provides an ursolate salt selected from ursolic acid diethanolamine salt and ursolic acid morpholine salt.

[0008] In a second aspect, the present application provides a composition comprising a ursolate selected from diethanolamine ursolate and morpholine ursolate, and other reagents.

[0009] In a third aspect, the present application provides the following method for a subject:

[0010]

[0011]

[0012]

[0013] The method comprises administering to the subject a ursolate selected from diethanolamine ursolate and morpholine ursolate, or administering to the subject a composition comprising a ursolate.

[0014] The following detailed description of various aspects of the present application in conjunction with the appended claims and drawings will make these features and other features and advantages of the present application obvious. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] The present application will be described below in conjunction with the drawings.

[0016] Figure 1A and 1B are micrographs of the skeletal muscle fibers of the quadriceps femoris of control and test mice, respectively, after 7 weeks of ad libitum intake of standard food (control) or standard food supplemented with 0.059% morpholine ursolate.

[0017] Figure 2A and 2B are micrographs of the adipocytes of the retroperitoneal fat pads of control and test mice, respectively, after 7 weeks of ad libitum intake of standard food (control) or standard food supplemented with 0.059% morpholine ursolate.

[0018] Figure 3 is a graph showing the results of a glucose tolerance test in control and test mice after 8 weeks of ad libitum intake of high-fat food (control) or high-fat food supplemented with 0.059% morpholine ursolate.

[0019] Figure 4 is a graph showing the results of a glucose tolerance test. DETAILED DESCRIPTION OF THE INVENTION

[0020] Various aspects of the present application and certain of its features, advantages and details will be described more fully below. Descriptions of well-known materials, devices, processing techniques, etc. are omitted so as not to unnecessarily obscure the present application.

[0021] In a first aspect, the present application provides a ursolate selected from diethanolamine ursolate and morpholine ursolate.

[0022] The arbutylates of the present application can be synthesized by techniques known in the art. The starting materials for the compounds of the present application can be obtained commercially, or they can be synthesized themselves using reagents and techniques known in the art. Non-limiting synthetic examples are described herein.

[0023] Ursolic acid, having the following structural formula:

[0024]

[0025] It is commercially available and is present in various plants from which it can be extracted, including apples, basil, bilberries, cranberries, elderflowers, mint, rosemary, lavender, oregano, thyme, hawthorn, and plums.

[0026] In some embodiments, the arbutylate of the present application is the diethanolamine salt of ursolic acid of the following formula:

[0027]

[0028] In some embodiments, the arbutylate of the present application is the morpholine salt of ursolic acid of the following formula:

[0029]

[0030] In a second aspect, the present application provides a composition comprising an arbutylate selected from the diethanolamine salt of ursolic acid and the morpholine salt of ursolic acid, and other reagents.

[0031] The other reagents are components present in the composition other than the arbutylate. Those of ordinary skill in the art are familiar with the various types of other reagents that can be included in the composition. For example, in some non-limiting embodiments, the other reagents include excipients, binders, diluents, carriers, other delivery carriers or systems, fillers, salts, buffers, preservatives, antioxidants, stabilizers, sweeteners, or flavoring agents.

[0032] In some embodiments, the other reagents are physiologically acceptable reagents. As used herein, a physiologically acceptable reagent is a reagent that does not exhibit significant toxicity to the intended subject at the administered dose. The term "physiologically acceptable reagent" includes substances generally regarded as safe (GRAS). The Food and Drug Administration lists GRAS substances in 21 CFR.§182 and 21 CFR§184 of the Code of Federal Regulations (CFR). The August 2017 editions of 21 CFR.§182 and 21 CFR§184 are incorporated herein by reference. The term "physiologically acceptable reagent" also includes pharmaceutically acceptable reagents.

[0033] As used herein, the term "pharmaceutically acceptable" means a component that is suitable for contact with the tissues of a subject (e.g., a human or other animal) within the scope of reasonable medical judgment, without undue toxicity, irritation, allergic response, etc., and is used in a manner commensurate with a reasonable benefit / risk ratio.

[0034] In some embodiments, the compositions of the present application are formulated for pharmaceutical use (i.e., are "pharmaceutical compositions"). A pharmaceutical composition includes a pharmaceutically acceptable carrier, adjuvant, or vehicle, which is "acceptable" in the sense of being compatible with the other ingredients of the formulation and is harmless to the recipient in the amounts commonly used in pharmaceuticals.

[0035] Pharmaceutically acceptable carriers, adjuvants, and vehicles that can be used in the pharmaceutical compositions of the present application include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins (e.g., human serum albumin), buffering substances (e.g., phosphates), glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes (e.g., protamine sulfate), disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polypropylene block polymers, polyethylene glycol, and lanolin.

[0036] In some embodiments, the composition is a pharmaceutical composition, a nutraceutical composition, a dietary composition, or a dietary supplement.

[0037] In some embodiments, the composition is in the form of, or includes, a non-naturally occurring oral delivery vehicle. Such delivery vehicles do not include naturally occurring carriers such as plants and fruits. Oral delivery vehicles include, but are not limited to, capsules, pills, tablets, cachets, syrups, foods, and beverages.

[0038] In some embodiments, the composition is in the form of one or more of the following dosage forms: tablets, cachets, capsules (e.g., soft elastic gelatin capsules), cachets, buccal tablets, lozenges, dispersions, suppositories, ointments, pastes (poultices), creams, powders, dressings, lotions, plasters, solutions, patches, aerosols (such as nasal sprays or inhalants), gels, liquid dosage forms suitable for oral or mucosal administration to a subject, including suspensions (e.g., aqueous or non-aqueous liquid suspensions, oil-in-water emulsions or water-in-oil liquid emulsions), solutions, and elixirs, liquid dosage forms suitable for parenteral administration to a patient; and sterile solid preparations (e.g., crystalline or amorphous solids) that can be reconstituted to provide a liquid dosage form suitable for parenteral administration to a subject.

[0039] In a third aspect, the present application provides a method for a subject:

[0040]

[0041]

[0042]

[0043]

[0044] The method comprises administering to a subject an ursolate selected from diethanolamine ursolate and morpholine ursolate, or administering to the subject a composition comprising an ursolate.

[0045] The various uses described above are described in the following references with respect to ursolic acid itself, the disclosures of which are hereby incorporated herein by reference in their entirety:

[0046] 1. Wozniak, L., Skapska, S., and Marszalek, K. (2015) Ursolic Acid - A Pentacyclic Triterpenoid with a Wide Spectrum of Pharmacological Activities. Molecules 20, 20614 - 20641

[0047] 2. Liu, J. (2005) Oleanolic acid and ursolic acid: research perspectives. J Ethnopharmacol 100, 92 - 94

[0048] 3. Sultana, N. (2011) Clinically useful anticancer, antitumor, and antiwrinkle agent, ursolic acid and related derivatives as medicinally important natural product. J Enzyme Inhib Med Chem 26, 616 - 642

[0049] 4. Liu, J. (1995) Pharmacology of oleanolic acid and ursolic acid. J Ethnopharmacol 49, 57 - 68

[0050] 5. Shanmugam, M.K., Dai, X., Kumar, A.P., Tan, B.K., Sethi, G., and Bishayee, A. (2013) Ursolic acid in cancer prevention and treatment: molecular targets, pharmacokinetics and clinical studies. Biochem Pharmacol 85, 1579 - 1587

[0051] 6. Kashyap, D., Tuli, H.S., and Sharma, A.K. (2016) Ursolic acid (UA): A metabolite with promising therapeutic potential. Life Sci 146, 201 - 213

[0052] 7. Lopez - Hortas, L., Perez - Larran, P., Gonzalez - Munoz, M.J., Falque, E., and Dominguez, H. (2018) Recent developments on the extraction and application of ursolic acid. A review. Food Res Int 103, 130 - 149

[0053] 8. Pironi, A.M., de Araujo, P.R., Fernandes, M.A., Salgado, H.R.N., and Chorilli, M. (2018) Characteristics, Biological Properties and Analytical Methods of Ursolic Acid: A Review. Crit Rev Anal Chem 48, 86 - 93

[0054] 9. Cargnin, S.T., and Gnoatto, S.B. (2017) Ursolic acid from apple pomace and traditional plants: A valuable triterpenoid with functional properties. Food Chem 220, 477 - 489

[0055] 10. Lee, S.U., Park, S.J., Kwak, H.B., Oh, J., Min, Y.K., and Kim, S.H. (2008) Anabolic activity of ursolic acid in bone: Stimulating osteoblast differentiation in vitro and inducing new bone formation in vivo. Pharmacol Res 58, 290 - 296

[0056] 11. Lu, J., Zheng, Y.L., Wu, D.M., Luo, L., Sun, D.X., and Shan, Q. (2007) Ursolic acid ameliorates cognition deficits and attenuates oxidative damage in the brain of senescent mice induced by D - galactose. Biochem Pharmacol 74, 1078 - 1090

[0057] 12. Wang, X.T., Gong, Y., Zhou, B., Yang, J.J., Cheng, Y., Zhao, J.G., and Qi, M.Y. (2018) Ursolic acid ameliorates oxidative stress, inflammation and fibrosis in diabetic cardiomyopathy rats. Biomed Pharmacother 97, 1461 - 1467

[0058] 13. Zhou, Y., Li, J.S., Zhang, X., Wu, Y.J., Huang, K., and Zheng, L. (2010) Ursolic acid inhibits early lesions of diabetic nephropathy. Int J Mol Med 26, 565 - 570

[0059] 14. Ling, C., Jinping, L., Xia, L., and Renyong, Y. (2013) Ursolic Acid provides kidney protection in diabetic rats. Curr Ther Res Clin Exp 75, 59 - 63

[0060] 15. Ding, H., Wang, H., Zhu, L., and Wei, W. (2017) Ursolic Acid Ameliorates Early Brain Injury After Experimental Traumatic Brain Injury in Mice by Activating the Nrf2 Pathway. Neurochem Res 42, 337 - 346

[0061] 16. Ma, J.Q., Ding, J., Zhang, L., and Liu, C.M. (2015) Protective effects of ursolic acid in an experimental model of liver fibrosis through Nrf2 / ARE pathway. Clin Res Hepatol Gastroenterol 39, 188 - 197

[0062] 17. Zhang, T., Su, J., Wang, K., Zhu, T., and Li, X. (2014) Ursolic acid reduces oxidative stress to alleviate early brain injury following experimental subarachnoid hemorrhage. Neurosci Lett 579, 12 - 17

[0063] 18. Hwang, T.L., Shen, H.I., Liu, F.C., Tsai, H.I., Wu, Y.C., Chang, F.R., and Yu, H.P. (2014) Ursolic acid inhibits superoxide production in activated neutrophils and attenuates trauma-hemorrhage shock-induced organ injury in rats. PLoS One 9, e111365

[0064] 19. Martinez-Abundis, E., Mendez-Del Villar, M., Perez-Rubio, K.G., Zuniga, L.Y., Cortez-Navarrete, M., Ramirez-Rodriguez, A., and Gonzalez-Ortiz, M. (2016) Novel nutraceutic therapies for the treatment of metabolic syndrome. World J Diabetes 7, 142 - 152

[0065] 20. Somova, L.O., Nadar, A., Rammanan, P., and Shode, F.O. (2003) Cardiovascular, antihyperlipidemic and antioxidant effects of oleanolic and ursolic acids in experimental hypertension. Phytomedicine 10, 115 - 121

[0066] 21. Somova, L.I., Shode, F.O., Ramnanan, P., and Nadar, A. (2003) Antihypertensive, antiatherosclerotic and antioxidant activity of triterpenoids isolated from Olea europaea, subspecies africana leaves. J Ethnopharmacol 84, 299 - 305

[0067] In some embodiments, the arbutylates described herein are particularly useful for the same indications asursolic acid itself.

[0068] As used herein, the term "subject" refers to an animal to which administration is targeted. In some embodiments, the subject is a human. In other embodiments, the subject is a non-human animal. In some embodiments, the subject is a non-rodent. In some embodiments, the subject is non-human and / or non-rodent. In some embodiments, the subject is a vertebrate and / or an amphibian. In some embodiments, the subject is a human, non-human primate, horse, pig, rabbit, dog, sheep, goat, cow, cat, guinea pig, hamster, ferret, fish, bird, or rodent. In some embodiments, the subject is a human or a domesticated animal. In some embodiments, the subject is a domesticated animal, such as a domestic fish, domestic crustacean, or domestic mollusk. In some embodiments, the domesticated animal is a poultry. In some embodiments, the poultry is selected from chickens, turkeys, ducks, and geese. In some embodiments, the domesticated animal is a livestock. In some embodiments, the livestock is selected from pigs, cows, horses, goats, bison, and sheep. In some embodiments, the livestock includes rodents. In other embodiments, the livestock does not include some or all rodents. In some embodiments, the livestock does not include mice and rats.

[0069] As used herein, the term "treatment" and its various forms (e.g., "treating") refer to the management (e.g., medical management) of a patient / subject aimed at curing, ameliorating, stabilizing, or preventing a disease, pathologic condition, or disorder. The term includes administering a compound and / or composition of the present application to a subject to treat, cure, alleviate, mitigate, alter, remedy, improve, modify, or affect a disease condition, disorder, or affliction, the symptoms of a disease condition, disorder, or affliction, or the susceptibility to a disease condition, disorder, or affliction. The term "treatment" and its various forms include uses for aesthetic and self-improvement purposes. For example, such uses include, but are not limited to, administering the disclosed compounds in nutraceuticals, medicated diets, energy bars, energy drinks, supplements (e.g., multivitamins). The term includes active treatment, i.e., treatment specifically directed to the improvement of a disease, pathologic condition, or disorder, and also includes etiological treatment, i.e., treatment directed to the elimination of the cause of the relevant disease, pathologic condition, or disorder. Additionally, the term includes palliative treatment, i.e., treatment aimed at alleviating symptoms rather than curing a disease, pathologic condition, or disorder; prophylactic treatment, i.e., treatment aimed at minimizing or partially or completely inhibiting the development of the relevant disease, pathologic condition, or disorder; and supportive treatment, i.e., treatment used to supplement another specific treatment directed to the improvement of the relevant disease, pathologic condition, or disorder. In various embodiments, the term encompasses any treatment of a subject and includes: (i) preventing the disease from occurring in a subject who may be predisposed to the disease but has not yet been diagnosed as having the disease; (ii) inhibiting the disease, i.e., preventing its progression; or (iii) alleviating the disease, i.e., causing the disease to regress. The term also includes precluding or preventing the onset, recurrence, or exacerbation of a disease condition, disorder, or affliction disclosed herein, as well as improving and / or reducing the occurrence of symptoms of a disease condition, disorder, or affliction.

[0070] Embodiments of the methods and uses of arbutyrates of the present application generally include administering an effective amount (e.g., a therapeutically effective amount) of the salt to a desired subject. As used herein, the term "therapeutically effective amount" or "effective amount" refers to an amount effective to cause a desired biological or medical response, including, when administered to a subject to treat a disease, an amount of a compound sufficient to effect treatment of such disease. The effective amount can vary depending on the compound, the condition and its severity, and the age, weight, etc. of the subject to be treated. The effective amount can be a single dose or multiple doses (e.g., it may be necessary to administer a single dose or multiple doses to reach the desired treatment endpoint). An effective amount can be considered effective if the desired or beneficial result can be achieved or obtained in combination with one or more other agents. The subjects in need of treatment include subjects at risk of developing a disease or disorder that requires treatment. As used herein, a "subject at risk" is a subject at risk of developing a disease or disorder to be treated. This can be indicated, for example, by one or more risk factors, which are measurable parameters known in the art as being associated with the development of a disease or disorder.

[0071] In some embodiments, before administering the salt of the present application to a subject, the subject has been diagnosed with a condition or disorder that requires treatment, or has been diagnosed with a condition or disorder. In some embodiments, before administering the salt of the present application to a subject, the subject has not been diagnosed with a condition or disorder. In some embodiments, the subject has not been diagnosed with cancer or diabetes. In other embodiments, the subject may have been diagnosed with a condition or disorder, such as cancer or diabetes.

[0072] As used herein, the term "diagnosed" means that a physical examination has been performed by a person skilled in the art (e.g., a physician), and the subject has been found to have a condition that can be diagnosed or treated by the arbutyrates, compositions or methods / uses of the present application disclosed herein. For example, "diagnosed with muscular dystrophy" means that a physical examination has been performed by a person skilled in the art (e.g., a physician), and the subject has been found to have a condition that can be diagnosed or treated by a compound or composition for promoting muscle health, promoting normal muscle function and / or promoting healthy muscle aging. As another example, "diagnosed as in need of promoting muscle health" means that a physical examination has been performed by a person skilled in the art (e.g., a physician), and the subject has been found to have a condition characterized by muscle atrophy or other diseases, where promoting muscle health, promoting normal muscle function and / or promoting healthy muscle aging would be beneficial to the subject. As discussed herein, such a diagnosis can refer to diseases such as muscular atrophy.

[0073] As used herein, phrases such as "diagnosed with a condition or disorder in need of treatment" refer to the selection of a subject based on the need for treatment of the condition. For example, based on an early diagnosis by a person skilled in the art, a subject can be diagnosed as having a disease in need of treatment (e.g., a disease associated with muscle atrophy), and then the disease can be treated. As another example, an object for consumption (e.g., fish, productive livestock, etc.) can be diagnosed as in need of treatment, such as to increase muscle mass in order to increase the average amount of meat that can be obtained per animal.

[0074] In some embodiments, the arbutyrate of the present application is administered at a dose of about 0.001 to about 500 mg / kg based on the body weight of the subject (e.g., 0.001, 0.002, 0.003, 0.004, 0.005, 0.006, 0.007, 0.008, 0.009, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.10, 0.11, 0.12, 0.13, 0.14, 0.15, 0.16, 0.17, 0.18, 0.19, 0.20, 0.21, 0.22, 0.23, 0.24, 0.25, 0.26, 0.27, 0.28, 0.29, 0.30, 0.31, 0.32, 0.33, 0.34, 0.35, 0.36, 0.37, 0.38, 0.39, 0.40, 0.41, 0.42, 0.43, 0.44, 0.45, 0.46, 0.47, 0.48, 0.49, 0.50, 0.51, 0.52, 0.53, 0.54, 0.55, 0.56, 0.57, 0.58, 0.59, 0.60, 0.61, 0.62, 0.63, 0.64, 0.65, 0.66, 0.67, 0.68, 0.69, 0.70, 0.71, 0.72, 0.73, 0.74, 0.75, 0.76, 0.77, 0.78, 0.79, 0.80, 0.81, 0.82, 0.83, 0.84, 0.85, 0.86, 0.87, 0.88, 0.89, 0.90, 0.91, 0.92, 0.93, 0.94, 0.95, 0.96, 0.97, 0.98, 0.99, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0, 9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, 9.9, 10.0, 10.1, 10.2, 10.3, 10.4, 10.5, 10.6, 10.7,10.8、10.9、11.0、11.1、11.2、11.3、11.4、11.5、11.6、11.7、11.8、11.9、12.0、12.1、12.2、12.3、12.4、12.5、12.6、12.7、12.8、12.9、13.0、13.1、13.2、13.3、13.4、13.5、13.6、13.7、13.8、13.9、14.0、14.1、14.2、14.3、14.4、14.5、14.6、14.7、14.8、14.9、15.0、16.1、16.2、16.3、16.4、16.5、16.6、16.7、16.8、16.9、17.0、17.1、17.2、17.3、17.4、17.5、17.6、17.7、17.8、17.9、18.0、18.1、18.2、18.3、18.4、18.5、18.6、18.7、18.8、18.9、19.0、19.1、19.2、19.3、19.4、19.5、19.6、19.7、19.8、19.9、20、21、22、23、24、25、26、27、28、29、30、31、32、33、34、35、36、37、38、39、40、41、42、43、44、45、46、47、48、49、50、51、52、53、54、55、56、57、58、59、60、61、62、63、64、65、66、67、68、69、70、71、72、73、74、75、76、77、78、79、80、81、82、83、84、85、86、87、88、89、90、91、92、93、94、95、96、97、98、99、100、101、102、103、104、105、106、107、108、109、110、111、112、113、114、115、116、117、118、119、120、121、122、123、124、125、126、127、128、129、130、131、132、133、134、135、136、137、138、139、140、141、142、143、144、145、146、147、148、149、150、151、152、153、154、155、156、157、158、159、160、161、162、163、164、165、166、167、168、169、170、171、172、173、174、175、176、177、178、179、180、181、182、183、184、185、186、187、188、189、190、191、192、193、194、195、196、197、198、199、200、201、202、203、204、205、206、207、208、209、210、211、212、213、214、215、216、217、218、219、220、221、222、223、224、225、226、227、228、229、230、231、232、233、234、235、236、237、238、239、240、241、242、243、244、245、246、247、248、249、250、251、252、253、254、255、256、257、258、259、260、261、262、263、264、265、266、267、268、269、270、271、272、273、274、275、276、277、278、279、280、281、282、283、284、285、286、287、288、289、290、291、292、293、294、295、296、297、298、299、300、301、302、303、304、305、306、307、308、309、310、311、312、313、314、315、316、317、318、319、320、321、322、323、324、325、326、327、328、329、330、331、332、333、334、335、336、337、338、339、340、341、342、343、344、345、346、347、348、349、350、351、352、353、354、355、356、357、358、359、360、361、362、363、364、365、366、367、368、369、370、371、372、373、374、375、376、377、378、379、380、381、382、383、384、385、386、387、388、389、390、391、392、393、394、395、396、397、398、399、400、401、402、403、404、405、406、407、407、408、409、410、411、412、413、414、415、416、417、418、419、420、421、422、423、424、425、426、427、428、429、430、431、432、433、434、435、436, 437, 438, 439, 440, 441, 442, 443, 444, 445, 446, 447, 448, 449, 450, 451, 452, 453, 454, 455, 456, 457, 458, 459, 460, 461, 462, 463, 464, 465, 466, 467, 468, 469, 470, 471, 472, 473, 474, 475, 476, 477, 478, 479, 480, 481, 482, 483, 484, 485, 486, 487, 488, 489, 490, 491, 492, 493, 494, 495, 496, 497, 498, 499 or 500 mg / kg), including any and all ranges and sub-ranges therein (e.g., 0.001 to 100 mg / kg, 0.01 to 75 mg / kg, 0.05 to 20 mg / kg, 0.1 to 10 mg / kg, etc.).

[0075] In some embodiments, the arbutyrate of the present application is administered in an amount of 1 to 2000 mg (such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267, 268, 269,270、271、272、273、274、275、276、277、278、279、280、281、282、283、284、285、286、287、288、289、290、291、292、293、294、295、296、297、298、299、300、301、302、303、304、305、306、307、308、309、310、311、312、313、314、315、316、317、318、319、320、321、322、323、324、325、326、327、328、329、330、331、332、333、334、335、336、337、338、339、340、341、342、343、344、345、346、347、348、349、350、351、352、353、354、355、356、357、358、359、360、361、362、363、364、365、366、367、368、369、370、371、372、373、374、375、376、377、378、379、380、381、382、383、384、385、386、387、388、389、390、391、392、393、394、395、396、397、398、399、400、401、402、403、404、405、406、407、408、409、410、411、412、413、414、415、416、417、418、419、420、421、422、423、424、425、426、427、428、429、430、431、432、433、434、435、436、437、438、439、440、441、442、443、444、445、446、447、448、449、450、451、452、453、454、455、456、457、458、459、460、461、462、463、464、465、466、467、468、469、470、471、472、473、474、475、476、477、478、479、480、481、482、483、484、485、486、487、488、489、490、490、491、492、493、494、495、496、497、498、499、500、1000、1010、1020、1030、1040、1050、1060、1070、1080、1090、1100、1110、1120、1130、1140, 1150, 1160, 1170, 1180, 1190, 1200, 1210, 1220, 1230, 1240, 1250, 1260, 1270, 1280, 1290, 1300, 1310, 1320, 1330, 1340, 1350, 1360, 1370, 1380, 1390, 1400, 1410, 1420, 1430, 1440, 1450, 1460, 1470, 1480, 1490, 1500, 1510, 1520, 1530, 1540, 1550, 1560, 1570, 1580, 1590, 1600, 1610, 1620, 1630, 1640, 1650, 1660, 1670, 1680, 1690, 1700, 1710, 1720, 1730, 1740, 1750, 1760, 1770, 1780, 1790, 1800, 1810, 1820, 1830, 1840, 1850, 1860, 1870, 1880, 1890, 1900, 1910, 1920, 1930, 1940, 1950, 1960, 1970, 1980, 1990 or 2000 mg), including any and all ranges and sub - ranges therein (e.g., 20 to 1900 mg, 50 to 1800 mg, 75 to 1700 mg, 100 to 1600 mg, etc.). Thus, in some embodiments where the arbutyrate is administered in the form of a composition, the composition will contain the above - mentioned amounts of the compound.,

[0076] In some embodiments, the administered amount of arbutyrate is greater than 50, 75, 100, 150, 200, 250, 300, 400, 500, 600, 750, 800, 900, 1000, 1100, 1200, 1300, 1400 or 1500 mg.

[0077] Examples

[0078] The present application will now be illustrated by way of example but not limited to the specific embodiments described in the following examples.

[0079] Sample Preparation - Ursolic Acid Diethanolamine Salt

[0080]

[0081] Arbutolic acid (1.0450 g) and diethanolamine (208.6 mg) were dissolved in THF:MeOH (10:1) and allowed to stand, then concentrated and dried to obtain a white solid: the diethanolamine salt of arbutolic acid (965.0 mg). This product was used as the diethanolamine salt of arbutolic acid in the tests described below.

[0082] Sample Preparation - Ursolic Acid Morpholine Salt

[0083]

[0084] ursolic acid (1.0715 g) was dissolved in 20 mL of THF. Morpholine (224 mg) was added and stirred for 1.5 h. 8 mL of hexane was added and the homogeneous solution became turbid. The salt was stirred at room temperature for 12 h. No substantial amount of salt formed. The solution was concentrated in vacuo and the stirred residue gave a white powder which was separated by filtration and dried in vacuo to give the morpholine salt of ursolic acid (867.1 mg). This product was used as the morpholine salt of ursolic acid in the tests described below.

[0085] Grip Strength Test Comparison of Ursolic Acid and Its Salts

[0086] Eight-week-old male C57BL / 6 mice of comparable body weight were randomly assigned to ad libitum consumption of standard chow (control) or standard chow supplemented with 0.050% ursolic acid, 0.200% ursolic acid, or one of the indicated ursolate salts: ursolic acid - ethanolamine salt, ursolic acid - tris(hydroxymethyl)aminomethane salt (hereinafter referred to as ursolic acid - tris salt), ursolic acid lysine salt, ursolic acid - diethanolamine salt, ursolic acid - morpholine salt, or ursolic acid - piperazine salt. The ursolic acid - diethanolamine salt and ursolic acid - morpholine salt were prepared as described above. Ursolic acid was commercially available and used as the starting material to prepare the other ursolate salts according to techniques known in the art similar to those described above. The doses of the ursolate salts were molar-matched to 0.050% ursolic acid (i.e., 0.057% ursolic acid - ethanolamine salt, 0.063% ursolic acid - tris salt, 0.066% ursolic acid lysine salt, 0.059% ursolic acid - morpholine salt, and 0.059% ursolic acid - piperazine salt), or to 0.048% ursolic acid (i.e., 0.060% ursolic acid - diethanolamine salt). After 6 weeks of the mice consuming the diets, the in vivo grip strength was measured. The results are shown in Table I below. The data are the mean ± SEM of 12 mice per group. One-way ANOVA with Dunnett's post-test showed **P < 0.01.

[0087] Table I

[0088]

[0089] As demonstrated by the data in Table I, not all of the ursolate salts showed a statistically significant improvement in strength. In fact, at the same dose, the efficacy of many of the salts may be less than that of the free acid. The ursolic acid - diethanolamine salt and ursolic acid - morpholine salt were unexpectedly more potent and effective than natural ursolic acid and other ursolate salts.

[0090] Ursolic Acid - Morpholine Salt - Grip Strength, Body Weight and Composition, and Skeletal Muscle Fiber Tests

[0091] Male C57BL / 6 mice, 8 weeks old and of comparable body weight, were randomly assigned to freely consume a standard diet (control) or a standard diet supplemented with 0.059% ursolic acid-morpholine salt. After 7 weeks of consuming the diet, grip strength was measured in vivo, the mice were weighed, body composition was evaluated by NMR, and the quadriceps muscles were dissected for histological analysis of the cross-sectional diameter of skeletal muscle fibers. The results are shown in Table II below. Body weight data, grip strength data, and NMR data (lean body mass, fat mass, lean%, and fat%) are the mean ± SEM of 14 - 15 mice per group. Muscle fiber diameter data per meal are the mean ± SEM of muscle fibers >4200. *P<0.05; **P<0.01; ***P<0.001

[0092] Table II

[0093] Control Ursolic Acid - Morpholine Salt Initial Body Weight (g) 26.1±0.4 26.3±0.3 Final Grip Strength (g) 165.5±3.3 181.9±2.4*** Final Body Weight (g) 35.6±0.8 34.6±1.0 Final Lean Body Mass (g) 19.6±0.3 20.8±0.3** Final Fat Mass (g) 10.2±0.9 7.8±1.2 Final Lean Body % 55.3±1.4 61.0±2.2* Final Fat % 28.0±2.1 21.4±2.9* Muscle Fiber Diameter (mm) 39.7±0.1 44.8±0.2***

[0094] The data in Table II demonstrate that ursolic acid-morpholine salt stimulates skeletal muscle hypertrophy, increases lean body mass, and reduces fat mass.

[0095] Ursolic Acid - Morpholine Salt - Skeletal Muscle Fiber Size Test

[0096] Male C57BL / 6 mice, 8 weeks old and of comparable body weight, were randomly assigned to freely consume a standard diet (control) or a standard diet supplemented with 0.059% ursolic acid-morpholine salt. After 7 weeks of the mice consuming the diet, the quadriceps muscles were dissected, fixed, sectioned, stained with hematoxylin and eosin, and photographed microscopically. Figure 1A This is a micrograph of the skeletal muscle fibers of the quadriceps muscle from the control group. Figure 1B This is a micrograph of the skeletal muscle fibers of the quadriceps muscle from the ursolic acid-morpholine salt group. Figure 1A Comparison with Figure 1B shows that ursolic acid-morpholine salt increases the size of skeletal muscle fibers.

[0097] Ursolic Acid - Morpholine Salt - Adipocyte Size Test

[0098] Male C57BL / 6 mice, 8 weeks old and of comparable body weight, were randomly assigned to freely consume a standard diet (control) or a standard diet supplemented with 0.059% ursolic acid-morpholine salt. After 7 weeks of the mice consuming the diet, the retroperitoneal fat pads were dissected, fixed, sectioned, stained with hematoxylin and eosin, and then photographed microscopically. Figure 2A This is a micrograph of the adipocytes of the retroperitoneal fat pad from the control group. Figure 2B This is a micrograph of the adipocytes of the retroperitoneal fat pad from the ursolic acid-morpholine salt group. Figure 2A Comparison with Figure 2B shows that ursolic acid-morpholine salt reduces the size of adipocytes.

[0099] Ursolic Acid - Morpholine Salt - Obesity and Blood Glucose Tests

[0100] Male C57BL / 6 mice at 8 weeks of age with comparable body weights were randomly assigned to freely consume a high-fat diet (control) or a high-fat diet supplemented with 0.059% ursolic acid-morpholine salt. Just before starting these diets, the mice were weighed and their body composition was analyzed by NMR. Nine weeks after the mice consumed the diets, the body weight and body components were re-evaluated, the retroperitoneal and epididymal fat pads were dissected and weighed, and non-fasting blood glucose was measured. The results are shown in Table III below. The data are the mean ± SEM of 15 mice per group. *P < 0.05; **P < 0.01.

[0101] Table III

[0102] Control Ursolic Acid - Morpholine Salt Initial Body Weight (g) 24.0±0.5 23.8±0.4 Initial Lean Body Mass (g) 16.5±0.4 16.7±0.4 Initial Fat Mass (g) 2.0±0.2 1.7±0.2 Initial Lean Body % 68.7±0.8 70.0±0.7 Initial Fat % 8.4±0.9 7.4±0.8 Final Body Weight (g) 41.6±1.0 38.9±1.2* Final Lean Body Mass (g) 20.3±0.6 21.3±0.6 Final Fat Mass (g) 16.5±0.9 12.8±1.4* Final Lean Body % 49.0±1.1 55.3±2.4* Final Fat % 39.4±1.6 31.8±3.1* Retroperitoneal Fat Pad Weight (mg) 929.4±52.0 731.8±86.9* Epididymal Fat Pad Weight (mg) 2257.0±84.6 1804.0±161.5** Blood Glucose (mg / dL) 198.1±6.1 177.7±9.9*

[0103] The data in Table III particularly demonstrate that ursolic acid-morpholine salt reduces obesity in diet-induced obese mice.

[0104] Ursolic Acid - Morpholine Salt - Glucose Tolerance Test

[0105] Male C57BL / 6 mice at 8 weeks of age with comparable body weights were randomly assigned to freely consume a high-fat diet (control) or a high-fat diet supplemented with 0.059% ursolic acid-morpholine salt. Eight weeks after the mice consumed the diets, they were fasted for 6 hours and then a glucose tolerance test was performed. The results are shown in Figure 3 , showing the glucose tolerance test result graph. The data are the mean ± SEM of 15 mice per group. It is Figure 3 apparent that the area under the curve (AUC) of the mice treated with ursolic acid-morpholine salt (31383 ± 1869) was significantly lower than that of the control mice (37122 ± 1339; P < 0.01), indicating that ursolic acid-morpholine salt significantly improved glucose tolerance (i.e., reduced glucose intolerance).

[0106] Ursolic Acid - Diethanolamine Salt - Body Weight and Composition Test

[0107] Male C57BL / 6 mice at 8 weeks of age with comparable body weights were randomly assigned to freely consume a standard diet (control), a standard diet supplemented with 0.060% ursolic acid-diethanolamine salt, or a standard diet supplemented with 0.180% ursolic acid-diethanolamine salt. Seven weeks after the mice consumed the diets, the mice were weighed and their body composition was evaluated by NMR. The results are shown in Table IV below. The data are the mean ± SEM of 15 mice per group. *P < 0.05.

[0108] Table IV

[0109]

[0110] The data in Table IV show that ursolic acid - diethanolamine salt increased lean body mass and decreased fat mass.

[0111] Direct Comparison of Ursolic Acid - Morpholine Salt with a Molar - Matched Dose of Ursolic Acid in Diet - Induced Obesity

[0112] Weight - matched 8 - week - old male C57BL / 6 mice were randomly assigned to freely consume a high - fat diet (control), a high - fat diet supplemented with 0.050% ursolic acid, or a high - fat diet supplemented with a molar - matched dose of ursolic acid - morpholine salt (0.059%). Just before starting these diets, the mice were weighed and body composition was analyzed by NMR. After 8 weeks of the mice consuming the diets, body weight and body composition were re - evaluated. Retroperitoneal and epididymal fat pads were dissected and weighed, and skeletal muscles (tibialis anterior, gastrocnemius, soleus, quadriceps, and triceps) were dissected and weighed. In each mouse, the total weight of the dissected skeletal muscles (the combined weight of bilateral tibialis anterior, gastrocnemius, soleus, quadriceps, and triceps) was normalized to the final body weight. Data are mean ± SEM for 10 mice per group. Different letters denote statistically significant differences (one - way ANOVA with Tukey's post - hoc comparison showed P < 0.05). Results are summarized in Table V.

[0113] Table V

[0114]

[0115] In Impaired Glucose Tolerance Caused by Obesity, the Direct Comparison

[0116] Weight - matched 8 - week - old male C57BL / 6 mice were randomly assigned to freely consume a high - fat diet supplemented with 0.050% ursolic acid or a high - fat diet supplemented with a molar - matched dose of ursolic acid - morpholine salt (0.059%). After 7 weeks of the mice consuming the diets, the mice were fasted for 6 hours and then a glucose tolerance test was performed. The results are shown in Figure 4 , which is the graph of the glucose tolerance test results. Data are mean ± SEM from 10 mice per group. The fasting blood glucose of mice treated with ursolic acid - morpholine salt (140.6 ± 7.8 mg / dL) was significantly lower than that of mice treated with ursolic acid (166.3 ± 3.7; P < 0.01). In addition, the area under the curve (AUC) of mice treated with ursolic acid - morpholine salt (32606 ± 2,471) was significantly lower than the AUC of mice treated with ursolic acid (40585 ± 1357; P < 0.01), indicating that ursolic acid - morpholine salt significantly reduced glucose intolerance relative to a molar - matched dose of ursolic acid.

[0117] As used herein, the terms "comprising" (and any form of comprising, such as "including" and "containing"), "having" (and any form of having, such as "has" and "with"), "including" (and any form of including, such as "includes" and "comprises"), "contains" (and any form of containing, such as "contains" and "includes"), and any other grammatical variants thereof, are open-ended linking verbs. As a result, a method or composition that "comprises", "has", "includes", or "contains" one or more steps or elements has those one or more steps or elements, but is not limited to only having those one or more steps or elements. Steps or elements. Similarly, steps of a method or elements of an article that "comprise", "have", "include", or "contain" one or more features have those one or more features, but are not limited to only having those one or more features.

[0118] As used herein, the terms "comprising", "having", "including", "containing" and their other grammatical variants encompass the terms "consisting of" and "consisting essentially of".

[0119] When used herein, the phrase "consisting essentially of" or its grammatical variants shall be regarded as designating the recited features, integers, steps or components, but excluding the addition of one or more further features, integers, steps, components or groups, provided that the additional features, integers, steps, components or groups do not materially alter the basic and novel features of the claimed composition or method.

[0120] Where ranges are referred to throughout the specification in connection with one or more items, each range is intended to represent a shorthand format for information, where the range is to be understood as covering every discrete point within the range as if it were fully set forth herein.

[0121] Although several aspects and embodiments of the present application have been described and depicted herein, those skilled in the art can effect alternative aspects and embodiments to achieve the same purpose. Accordingly, the present disclosure and the appended claims are intended to cover all such other and alternative aspects and embodiments that fall within the true spirit and scope of the present application.

Claims

1. A bearberry salt, selected from diethanolamine salt of ursolic acid and morpholine salt of ursolic acid, wherein the salt is the diethanolamine salt of ursolic acid of the following formula: ， wherein the salt is the morpholine salt of ursolic acid of the following formula: 。 2. The bearberry salt according to claim 1, wherein the salt is the morpholine salt of ursolic acid.

3. A composition, comprising the salt according to claim 1 and other reagents.

4. The composition according to claim 3, comprising 25 to 1500 mg of the salt.

5. The composition according to claim 3, comprising 50 to 1000 mg of the salt.

6. The composition according to claim 3, wherein the other reagents comprise a physiologically acceptable carrier.

7. The composition according to claim 3, wherein the composition is a pharmaceutical composition, a nutraceutical composition, a dietary composition or a dietary supplement.

8. The composition according to claim 7, comprising 25 to 1500 mg of the salt.

9. The composition according to any one of claims 3 - 8, wherein the salt is the morpholine salt of ursolic acid.

10. Use of an effective amount of the salt according to claim 1 in the manufacture of a medicament for the following purposes in a subject: (ii) Treating skeletal muscle atrophy; (iv) Treating cachexia; (xvii) Treating obesity; (xviii) Treating diabetes; (xx) Treating prediabetes; (xxi) Treating metabolic syndrome; (xxii) Treating insulin resistance; (xxiv) Treating hypercholesterolemia; (xxvi) Treating hypertriglyceridemia; or (xxix) Treating non - alcoholic fatty liver disease (NAFLD).

11. The use according to claim 10, wherein, The effective amount of the salt is 25 to 1500 mg.

12. The use according to claim 10, for the manufacture of a medicament for the following purposes: (ii) Treating skeletal muscle atrophy; (iv) Treating cachexia; or (xiii) Treating obesity.

13. The use according to claim 12, wherein, The effective amount of the salt is 25 to 1500 mg.

14. The use according to any one of claims 10 - 13, wherein the salt is the morpholine salt of ursolic acid.

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