METAP2 inhibitors and methods for treating obesity

Abstract

To provide modified or polymer conjugated MetAP2 inhibitors.SOLUTION: The present invention relates to methods of preventing, inducing, causing or increasing weight loss, treating obesity and / or treating metabolic syndrome utilizing the modified or polymer conjugated MetAP2 inhibitors. The present invention also relates to methods of improving insulin sensitivity and glycemic control, reducing insulin levels and / or improving leptin sensitivity utilizing the modified or polymer conjugated MetAP2 inhibitors.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Patent Application No. 61 / 810,468, filed April 10, 2013, and U.S. Provisional Patent Application No. 61 / 925,918, filed January 10, 2014, the contents of each of which are incorporated herein by reference in their entirety. [Background technology]

[0002] Obesity is a chronic disease and a major health concern in modern society. According to the Centers for Disease Control and Prevention (CDC), the United States is in the midst of an obesity epidemic. Approximately 65% ​​of adults in the United States are overweight, 30% are obese, and over 5 million adults are classified as morbidly obese. Another 10 million people are closer to that threshold and at risk for obesity-related health problems. The problem is only growing, with childhood and adolescent obesity having doubled in the past 20 years.

[0003] Existing obesity treatments include diet and exercise; very low calorie diets; behavioral therapy; pharmacotherapy including appetite suppressants, thermogenic drugs, and food absorption inhibitors; mechanical devices such as jaw wiring, waist cords, and balloons; and surgery. However, these existing treatments are not very effective. Adherence to energy-restricted diets is problematic and generally ineffective, and pharmacotherapy is only marginally effective in long-term weight management. In many cases, toxicity and side effects prevent the development of potential anti-obesity drug candidates. Metabolic syndrome (Sutherland et al., Metabolic Syndrome and Related Disorders 2:82-104 (2004); Esposito et al., Nutr. Metab. Cardiovasc. Dis. 14:228-232 (2004)) is associated with obesity and is characterized by a constellation of metabolic risk factors including: 1) abdominal obesity (excess adipose tissue in and around the abdomen); 2) atherogenic dyslipidemia (high triglycerides; low HDL cholesterol and high LDL cholesterol); 3) elevated blood pressure; 4) insulin resistance or impaired glucose tolerance; 5) a prothrombotic state (e.g., high fibrinogen or plasminogen activator inhibitor-1 in the blood); and 6) a proinflammatory state (e.g., elevated CRP in the blood). Metabolic syndrome is becoming increasingly prevalent in developed countries and is closely associated with the risk of coronary heart disease (Malik et al., Circulation 110:1245-1250 (2004); Irabarren et al., J. Am. Coll. Cardiol. 48:1800-1807 (2006)).

[0004] Cardiometabolic syndrome includes obesity-related metabolic disorders and atherosclerosis. Cardiometabolic disorders also promote arterial and valvular calcification, which can result in acute myocardial infarction and aortic stenosis, which are devastating clinical complications. Diabetes also causes chronic kidney disease, which also leads to cardiovascular ectopic calcification and acute myocardial infarction. Collectively, several key components of cardiometabolic syndrome, which develop through interrelated mechanisms, promote each other through local or systemic inflammation. Furthermore, lack of patient adherence to prescribed drug therapy represents a huge challenge for the global health care community. In the United States alone, avoidable medical expenditures were estimated at $300 billion in 2009. With blockbuster drugs expiring on patent, exhausting distribution channels, and cost containment by users, closing this adherence gap is a "must do" for pharmaceutical companies.

[0005] Thus, there is a need for novel compounds and methods for causing, inducing, and / or increasing weight loss, and for treating obesity and metabolic syndrome. The present invention addresses these needs. Summary of the Invention [Means for solving the problem]

[0006] The present invention provides a method for inducing or causing weight loss in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of at least one compound of the present invention to induce or cause weight loss.In some embodiments, the subject is overweight or obese.In some embodiments, inducing or causing weight loss is increasing weight loss.

[0007] The present invention also provides methods for treating obesity, metabolic syndrome and / or related comorbidities in a subject in need thereof, comprising administering to the subject on a reasonable schedule a therapeutically effective amount of at least one compound of the present invention, or a pharma- ceutically acceptable salt, prodrug, metabolite, analog or derivative thereof, to treat or ameliorate these diseases and conditions.

[0008] The present invention also provides methods for improving insulin sensitivity and glycemic control, reducing insulin levels, and / or improving leptin sensitivity in a subject in need thereof, comprising administering to the subject on a rational schedule a therapeutically effective amount of at least one compound of the present invention, or a pharma- ceutically acceptable salt, prodrug, metabolite, analog or derivative thereof, to treat or ameliorate these diseases and conditions.

[0009] 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. As used herein, the singular includes the plural unless the context clearly indicates otherwise. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference. References cited herein are not admitted to be prior art to the claimed invention. In case of conflict, the present specification, including definitions, will control. Additionally, the materials, methods, and examples are illustrative only and are not intended to be limiting. [Brief description of the drawings]

[0010] [Figure 1] 1 is a graph showing the change in body weight over time following administration of a compound of the present invention. [Diagram 2] 1 is a graph showing average food intake by week following administration of a compound of the invention. [Diagram 3] 1 is a graph showing the relationship between total fat mass and body weight after administration of a compound of the present invention. [Figure 4] 1 is a graph showing blood glucose levels after glucose challenge during treatment of mice using compounds of the present invention. [Diagram 5] FIG. 1 is a graph showing the change in body weight over time following administration of various doses of a compound of the invention on a q4d dosing schedule. [Figure 6] 1 is a graph showing the change in total cholesterol, triglycerides, HDL cholesterol and LDL cholesterol as a function of dose level using compounds of the invention at the end of a 32 day study. [Figure 7] 1 is a graph showing the change in body weight over time after administration of various doses of a compound of the invention on a q7d dosing schedule to rats. [Figure 8] 1 is a graph showing the change in body weight in rats treated with single doses of various test agents. [Figure 9] 1 is a graph showing plasma concentrations of a compound of the invention over time based on administration of two different compounds. [Figure 10] 1 is a graph showing the change in body weight over time following administration of various compounds of the invention. [Figure 11] 1 is a graph showing body weight change in male Levin rats fed a high fat diet following administration of various compounds of the invention. [Figure 12] 1 is a graph showing weight loss versus fumagillol exposure in DIO rats following administration of various compounds of the invention. [Figure 13] 1 is a graph showing changes in insulin levels in male Levin DIO rats fed a high fat diet following administration of various compounds of the invention. [Figure 14] 1 is a graph showing insulin levels during an oral glucose challenge (OGTT) following administration of various compounds of the invention. [Figure 15]1 is a graph showing the reduction in blood glucose in DIO rats during an oral glucose challenge (OGTT) following administration of various compounds of the invention. [Figure 16] FIG. 1 is a graph showing HOMA-ir product during OGTT in DIO rats after administration of various compounds of the invention. [Figure 17] FIG. 1 is a graph showing weekly food intake following administration of various compounds of the invention. [Figure 18] 1 is a graph showing the change in leptin levels from baseline following administration of various compounds of the invention. [Figure 19] 1 is a graph showing body weight change in DIO mice following administration of various compounds of the invention on Q4D and Q8D schedules. [Figure 20] 1 is a graph showing weight loss following administration of CKD-732 on a Q2D and Q4D schedule. [Figure 21] 1 is a graph showing the reduction in food intake following administration of various compounds of the invention. [Figure 22] FIG. 1 is a graph showing significantly decreased insulin levels during ipGTT in male C57B16 mice fed a high fat diet for 25 weeks following administration of various compounds of the invention. [Diagram 23] 1 is a graph showing insulin AUC during glucose challenge in male DIO mice following administration of various compounds of the invention. [Figure 24] 1 is a graph showing blood glucose levels in male C57B16 mice fed a high fat diet following administration of various compounds of the invention. [Diagram 25] FIG. 1 is a graph showing HOMA product during ipGTT in C57B16 mice fed a high fat diet after administration of various compounds of the invention. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0011] Compounds of the Invention The present invention provides a drug conjugate composition comprising a modified active moiety, a conjugate moiety, and a cleavable linker, where cleavage of the linker occurs substantially in the target tissue to produce a modified active moiety that has reduced efflux from the target tissue compared to the unmodified active moiety. The present invention also provides a composition comprising the modified active moiety.

[0012] The conjugate moiety used depends on the biological requirements, such as the pharmacokinetic and pharmacodynamic properties of the active moiety, as well as knowledge of the disease state, as well as the physicochemical properties of both the conjugate moiety and the active moiety. A person skilled in the art will be able to select an appropriate conjugate moiety based on the above considerations. The conjugate moiety is used to deliver small molecule active moieties or larger molecule active moieties, such as proteins, peptides or oligonucleotides.

[0013] The conjugate moiety improves the delivery of the active moiety to the target. The conjugate moiety is selected to maximize the bioavailability of the active moiety; optimize the onset, duration and delivery rate of the active moiety; and maintain the concentration of the active moiety in the therapeutic range in the target tissue for as long as required for effective treatment. The conjugate moiety can also help minimize adverse side effects of the active moiety. Thus, the conjugate moiety prolongs the pharmacological activity of the active moiety, stabilizes labile active moieties from chemical and proteolytic degradation, minimizes side effects, increases solubility, and delivers the active moiety to specific cells or tissues.

[0014] Other properties of the conjugate moiety to be considered are that the conjugate moiety has minimal immunogenicity and toxicity or is non-immunogenic and non-toxic. The molecular weight of the conjugate moiety should be large enough to avoid rapid removal by renal ultrafiltration and small enough to prevent undesirable accumulation in the body. In some embodiments, the conjugate moiety is hydrophilic and biodegradable. Non-biodegradable conjugate moieties are also suitable in the compositions and methods of the present invention. The conjugate moiety should be capable of carrying the required amount of the active moiety and preventing premature metabolism of the active moiety during transport to the target tissue.

[0015] Preferred conjugates include all forms of polymers, i.e., synthetic and natural product-related polymers, including peptides, polysaccharides, polynucleic acids, antibodies and aptamers. In a preferred embodiment, the conjugate is a synthetic polymer. Preferred polymers of the present invention are described in U.S. Pat. Nos. 4,997,878 to Bock et al., 5,037,883 to Kopecek et al., 5,258,453 to Kopecek et al., 6,464,850 to Zhang et al., and 6,803,438 to Brocchini et al., each of which is incorporated by reference in its entirety. Additional preferred polymers are described in Subr et al., J Controlled Release, 18, 123-132 (1992). In some embodiments, the synthesis method of the polymer may result in the coupling of two or more polymer chains, which may increase the weight average molecular weight of the polymer conjugate. It is further understood that when this coupling occurs, the bond is biodegradable.

[0016] The active moiety may be any compound or molecule that produces a therapeutic effect in a subject. In some embodiments, the compound or molecule has a molecular weight of 2000 daltons or less, 1500 daltons or less, 1000 daltons or less, 500 daltons or less, or 250 daltons or less. In some embodiments, the compound or molecule is a MetAP2 inhibitor. In some embodiments, the compound or molecule is fumagillin, fumagillol, or an analog, derivative, salt, or ester thereof. The compound or molecule selected depends on the condition or disease being treated. In some embodiments, more than one active moiety may be used. In some embodiments, an active moiety and an inactive "capping" moiety may be used. In some embodiments, the condition being treated is obesity. In the compositions of the present invention, the conjugate moiety is linked to the active moiety via a linker. Any linker structure known in the art may be used to link the modified active moiety to the conjugate moiety. The linker used depends on the physiological condition of the target tissue, the properties of the active moiety being optimized, and the cleavage mechanism. D'Souza et al. review various linkers, including those that function via proteolytic cleavage: "Release from Polymeric Prodrugs: Linkages and Their Degradation" J. Pharm. Sci., 93, 1962-1979 (2004). Blencoe et al. describe various self-immolative linkers: "Self-immolative linkers in polymeric delivery systems" Polym. Chem. 2, 773-790 (2011). Ducry et al. review linkers in Bioconj. Chem. 21, 5-13 (2010) "Antibody-Drug Conjugates: Linking Cytotoxic Payloads to Monoclonal Antibodies".Peptide linkers suitable for cleavage by matrix metalloproteins (MMPs) are described in Chau et al., “Antitumor efficacy of a novel polymer-peptide-drug conjugate in human tumor xenograft models,” Int. J. Cancer 118, 1519-1526 (2006), and in Chau et al., U.S. Patent Application Publication No. 2004 / 0116348. Other linker chemistries suitable for the compositions of the invention are set forth in Shiose et al., Biol. Pharm. Bull. 30(12) 2365-2370 (2007); Shiose et al., Bioconjugate Chem. 20(1) 60-70 (2009); U.S. Pat. Nos. 7,553,816 to Senter; 7,223,837 to De Groot; 6,759,509 to King; 6,835,807 to Susaki; 6,436,912 to Susaki; and 7,943,569 to Gemeinhart.

[0017] In some embodiments, the linker is a peptide linker. Preferred peptide linkers are described in U.S. Patent No. 6,835,807 to Susaki et al., U.S. Patent No. 6,291,671 to Inoue et al., U.S. Patent No. 6,811,996 to Inoue et al., U.S. Patent No. 7,041,818 to Susaki et al., U.S. Patent No. 7,091,186 to Senter et al., and U.S. Patent No. 7,553,816 to Senter et al., each of which is incorporated by reference in its entirety. Additional preferred peptides and their truncations are described in Shiose et al., Biol.Pharm.Bull.30(12)2365-2370(2007) and Shiose et al., Bioconjugate Chem.20(1)60-70(2009). Peptide linkers suitable for cleavage by matrix metalloproteins (MMPs) are described in Chau et al., “Antitumor efficacy of a novel polymer-peptide-drug conjugate in human tumor xenograft models” Int. J. Cancer 118, 1519-1526 (2006), and Chau et al., U.S. Patent Application Publication No. 2004 / 0116348.

[0018] The linker may be cleaved by any mechanism known in the art. For example, the linker may be designed for proteolytic or intracellular proteolytic cleavage. In some embodiments, the linker is designed such that there is no cleavage of the linker in plasma or the cleavage rate is very low in plasma. Preferred linker structures are described in more detail below.

[0019] In some embodiments, the linker is structured to be preferentially cleaved in diseased tissue. Since hydrolases exist in both normal and diseased tissues, the linker should be cleaved by hydrolases that are more active and / or more abundant in diseased tissue. For example, tumors generally have upregulated metabolic rates and overexpress proteases, including cathepsins in particular. The role of upregulation and proteases in cancer is described in Mason et al., Trends in Cell Biology 21,228-237 (2011).

[0020] In one embodiment, a class of modified active moieties are those that bind irreversibly to their targets; i.e., after release from the complex, the active moiety is covalently bound to the biochemical target. Once bound, the active moiety cannot diffuse or be transported out of the cell. In the case of irreversible binding, the rate of binding of the small molecule to the target, K, must be met for targeting to occur. rev1 is the rate of small molecule efflux, k sm-1 A high rate of efflux relative to small molecule binding would establish an equilibrium of the small molecule between the plasma and intracellular compartments, eliminating the advantage of intracellular versus extracellular delivery.

[0021] In another embodiment, a class of modified active moieties are those that bind reversibly to their targets. In the case of reversible binding, the equilibrium constant for small molecule binding to a target, K=k rev1 / k rev-1 should be large, and the "on-rate", k rev1 is the rate of small molecule efflux, k sm-1should be high relative to the rate of small molecule binding. A high rate of efflux relative to the rate of small molecule binding will establish an equilibrium of the small molecule between the plasma and intracellular compartments, eliminating the advantage of intracellular delivery over extracellular delivery. Such a relationship is shown diagrammatically below, where: [PC] = concentration of polymer conjugate; [SM] = concentration of free small molecule; plasma = plasma concentration; icell = concentration within a cell; icell-target = small molecule reversibly bound to a target within a cell; and inactive = inactive metabolite of the small molecule. In one embodiment, k rev-1 When =0, the moiety irreversibly binds to the target.

[0022] [ka]

[0023] In another embodiment, the class of modified active moieties are those that have very high equilibrium constants and high "on-rates" relative to efflux. In another embodiment, the class of modified active moieties are those that undergo intracellular metabolism at high rates relative to efflux.

[0024] In some embodiments, the modification to the active moiety is achieved by using a linker that has a structure such that upon cleavage, a fragment of the linker remains attached to the active moiety. The fragment can either change the molecular weight, hydrophobicity, polar surface area, or charge of the active moiety, thereby generating a modified active moiety that has reduced efflux from target cells compared to the unmodified active moiety. For example, a MetAP2 inhibitory active moiety can be coupled via a linker as described herein, resulting in a complex that, upon cleavage of the linker, generates an active moiety (modified active moiety) with a fragment of the linker attached thereto. The modified active moiety as described herein can have reduced efflux from cells compared to the unmodified active moiety, resulting in a modified active moiety with better efficacy relative to the parent small molecule, and better efficacy relative to the parent small molecule, and a better pharmacokinetic profile.

[0025] The present invention relates to the following:

[0026] [ka]

[0027] wherein, independently at each occurrence, R 4 is hydrogen or C 1 -C 6 R is alkyl; 5 is hydrogen or C 1 -C 6 R is alkyl; 6 is C 2 -C 6 hydroxyalkyl; Z is -NH-AA 1 -AA 2 -AA 3 -AA 4 -AA 5 -AA 6 -C(O)-L or -NH-AA 1 -AA 2 -AA 3 -AA 4 -AA 5 -AA 6 -C(O)-QXYC(O)-W;AA 1 is glycine, alanine, or H 2 N(CH 2 ) mCO 2 H, m is 2, 3, 4 or 5; AA 2 is a bond or alanine, cysteine, aspartic acid, glutamic acid, phenylalanine, glycine, histidine, isoleucine, lysine, leucine, methionine, asparagine, proline, glutamine, arginine, serine, threonine, valine, tryptophan, or tyrosine; AA 3 is a bond or alanine, cysteine, aspartic acid, glutamic acid, phenylalanine, glycine, histidine, isoleucine, lysine, leucine, methionine, asparagine, proline, glutamine, arginine, serine, threonine, valine, tryptophan, or tyrosine; AA 4is a bond or alanine, cysteine, aspartic acid, glutamic acid, phenylalanine, glycine, histidine, isoleucine, lysine, leucine, methionine, asparagine, proline, glutamine, arginine, serine, threonine, valine, tryptophan, or tyrosine; AA 5 is a bond or glycine, valine, tyrosine, tryptophan, phenylalanine, methionine, leucine, isoleucine, or asparagine; AA 6 is a bond or alanine, asparagine, citrulline, glutamine, glycine, leucine, methionine, phenylalanine, serine, threonine, tryptophan, tyrosine, valine, or H 2 N(CH 2 ) mCO 2 H, m is 2, 3, 4 or 5; L is -OH, -O-succinimide, -O-sulfosuccinimide, alkoxy, aryloxy, acyloxy, aroyloxy, alkoxycarbonyloxy, aryloxycarbonyloxy, -NH 2 , -NH(C 2 -C 6 hydroxyalkyl), halide or perfluoroalkyloxy; Q is NR, O, or S; X is M-(C(R) 2 ) p -MJM-(C(R) 2 ) p -MV; M is a bond or C(O); J is a bond or ((CH 2 ) q Q) r , C 5 -C 8 Y is NR, O, or S; R is hydrogen or alkyl; V is a bond or:

[0028] [ka]

[0029] ;R 9is alkyl, aryl, aralkyl, or a bond; or R 9 forms a heterocycle together with Y; R 10 is an amide or a bond; R 11 is hydrogen or alkyl; W is a MetAP2 inhibitor moiety or alkyl; x ranges from 1 to about 450; y ranges from 1 to about 30; n ranges from 1 to about 50; p ranges from 0 to 20; q is 2 or 3; and r is 1, 2, 3, 4, 5, or 6. The present invention provides a conjugate having a linker having the structure:

[0030] In one embodiment, R 4 is C 1 -C 6 In one embodiment, R 4 is methyl. In one embodiment, R 5 is C 1 -C 6 In one embodiment, R 5 is methyl. In one embodiment, R 6 is 2-hydroxyethyl, 2-hydroxypropyl, or 3-hydroxypropyl. 6 is 2-hydroxypropyl.

[0031] In some embodiments, the compound has a molecular weight of less than about 60 kDa. In other embodiments, the molecular weight is less than about 45 kDa. In other embodiments, the molecular weight is less than about 35 kDa.

[0032] In some embodiments, the ratio of x to y is in the range of about 30:1 to about 3:1. In other embodiments, the ratio of x to y is in the range of about 19:2 to about 7:2. In some embodiments, the ratio of x to y is in the range of about 9:1 to about 4:1. In some embodiments, the ratio of x to y is about 11:1. In some embodiments, the ratio of x to y is about 9:1. In some embodiments, the ratio of x to y is about 4:1.

[0033] In one embodiment, Z is -hydrogen -AAi-AA 2 -AA 3 -AA 4 -AA 5 -AA 6 In some embodiments, L is -C(O)-L. In some embodiments, L is methoxy, ethoxy, pentafluorophenyloxy, phenyloxy, acetoxy, fluoride, chloride, methoxycarbonyloxy; ethoxycarbonyloxy, phenyloxycarbonyloxy, 4-nitrophenyloxy, trifluoromethoxy, pentafluoroethoxy, or trifluoroethoxy. In some embodiments, L is 4-nitrophenyloxy.

[0034] In one embodiment, Z is -hydrogen-AA 1 -AA 2 -AA 3 -AA 4 -AA 5 -AA 6 -C(O)-QXYC(O)-W. In one embodiment, AA 1 is glycine. In one embodiment, AA 2 is glycine. In one embodiment, AA 3 is glycine. In one embodiment, AA 4 is glycine or phenylalanine. In one embodiment, AA 5 is leucine, phenylalanine, valine, or tyrosine. 6 is asparagine, citrulline, glutamine, glycine, leucine, methionine, threonine, or tyrosine. 5 -AA 6 are Leu-Cit, Leu-Gin, Leu-Gly, Leu-Leu, Leu-Met, Leu-Thr, Phe-Cit, Phe-Gln, Phe-Leu, Phe-Met, Ph e-Thr, Val-Asn, Val-Cit, Val-Gln, Val-Leu, Val-Met, Val-Thr, Tyr-Cit, Tyr-Leu, or Tyr-Met. In certain embodiments, A.A. 1 , A.A. 3and A.A. 5 is glycine, valine, tyrosine, tryptophan, phenylalanine, methionine, leucine, isoleucine, or asparagine. 2 , A.A. 4 and A.A. 6 is glycine, asparagine, citrulline, glutamine, glycine, leucine, methionine, phenylalanine, threonine, or tyrosine. 2 is a bond; and AA 3 is a bond. In some embodiments, AA 1 is glycine; AA 4 is phenylalanine; AA 5 is leucine; and AA 6 is glycine.

[0035] In one embodiment, W is:

[0036] [ka]

[0037] [ka]

[0038] [ka]

[0039] [ka]

[0040] [ka]

[0041] and

[0042] In the formula, R 2is -OH or methoxy; and R 3 is hydrogen, -OH or methoxy.

[0043] In one embodiment, W is:

[0044] [ka]

[0045] It is.

[0046] In one embodiment, W is:

[0047] [ka]

[0048] It is.

[0049] In some embodiments, Q is NR. In other embodiments, Q is S.

[0050] In some embodiments, J is NR. In other embodiments, J is ((CH 2 ) q Q) r In another embodiment, J is C. 5 -C 8 In some embodiments, J is aryl.

[0051] In some embodiments, Y is NR. In other embodiments, Y is S.

[0052] In one embodiment, -QXY is:

[0053] [ka]

[0054] [ka]

[0055] and;

[0056] V is:

[0057] [ka]

[0058] , or a bond; R 12 is hydrogen or Me; R 12 is R 14 Together with R, they form a piperidine ring; 11 is hydrogen or Me; and R 13 is R 12 Together with

[0059] In one embodiment, -QXY- is:

[0060] [ka]

[0061] It is.

[0062] In one embodiment, -QXY- is:

[0063] [ka]

[0064] It is.

[0065] In one embodiment, -QXY- is:.

[0066] [ka]

[0067] It is.

[0068] In one embodiment, -QXY- is:

[0069] [ka]

[0070] In one embodiment, -QXY- is:

[0071] [ka]

[0072] It is.

[0073] In one embodiment, R and R 5 is methyl; R 6 is 2-hydroxypropyl; Z is -NH-AA 1 -AA 2 -AA 3 -AA 4 -AA 5 -AA 6 -C(O)-QXYC(O)-W;AA 1 is glycine; AA 2 is a bond; AA 3 is a bond; AA 4 is phenylalanine; AA 5 is leucine; AA 6 is glycine; -QXY- is:

[0074] [ka]

[0075] and W is:

[0076] [ka]

[0077] It is.

[0078] In one embodiment, R 4 and R 5 is methyl; R 6 is 2-hydroxypropyl; Z is -NH-AA 1 -AA 2 -AA 3 -AA 4 -AA 5 -AA 6 -C(O)-QXYC(O)-W;AA 1 is glycine; AA 2 is a bond; AA 3 is a bond; AA 4 is phenylalanine, AA 5 is leucine, AA 6 is glycine; -QXY- is:

[0079] [ka]

[0080] and W is:

[0081] [ka]

[0082] It is.

[0083] In one embodiment, R 4 and R 5 is methyl; R 6 is 2-hydroxypropyl; Z is -NH-AA 1 -AA 2 -AA 3 -AA 4 -AA 5 -AA 6 -C(O)-QXYC(O)-W;AA 1 is glycine; AA 2 is a bond; AA 3 is a bond; AA 4is phenylalanine; AA 5 is leucine; AA 6 is glycine; -QXY- is:

[0084] [ka]

[0085] and W is:

[0086] [ka]

[0087] It is.

[0088] In one embodiment, -QXY- is represented by the following scheme:

[0089] [ka]

[0090] As shown in Figure 1, it is a self-immolative linker that releases a MetAP2 inhibitor in the form of a carbamate derivative.

[0091] Another aspect of the invention provides conjugates having a linker having the structure ZQXYC(O)-W, wherein, independently in each occurrence, Z is H. 2 N-AA 2 -AA 3 -AA 4 -AA 5 -AA 6 -C(O)- or hydrogen; AA 2 is a bond or alanine, cysteine, aspartic acid, glutamic acid, phenylalanine, glycine, histidine, isoleucine, lysine, leucine, methionine, asparagine, proline, glutamine, arginine, serine, threonine, valine, tryptophan, or tyrosine; AA 3is a bond or alanine, cysteine, aspartic acid, glutamic acid, phenylalanine, glycine, histidine, isoleucine, lysine, leucine, methionine, asparagine, proline, glutamine, arginine, serine, threonine, valine, tryptophan, or tyrosine; AA 4 is a bond or alanine, cysteine, aspartic acid, glutamic acid, phenylalanine, glycine, histidine, isoleucine, lysine, leucine, methionine, asparagine, proline, glutamine, arginine, serine, threonine, valine, tryptophan, or tyrosine; AA 5 is a bond, alanine, cysteine, glycine, isoleucine, leucine, methionine, phenylalanine, valine, tryptophan, or 6 is alanine, asparagine, citrulline, glutamine, glycine, leucine, methionine, phenylalanine, serine, threonine, tryptophan, tyrosine, valine or H 2 N(CH 2 ) mCO 2 H, m is 2, 3, 4 or 5, Q is NR, O, or S; X is M-(C(R) 2 ) P -MJM-(C(R) 2 ) P -MV; M is a bond or C(O); J is a bond or ((CH 2 ) q Q) r , C 5 -C 8 Y is NR, O, or S; R is hydrogen or alkyl; V is a bond or:

[0092] [ka]

[0093] ;R 9 is alkyl, aryl, aralkyl, or a bond; or R 9forms a heterocycle together with Y; R 10 is an amide or a bond, R 11 is hydrogen or alkyl, W is a MetAP2 inhibitor moiety, p is 0-20; q is 2 or 3; and r is 1, 2, 3, 4, 5, or 6.

[0094] In one embodiment, Z is H 2 N-AA 5 -AA 6 In one embodiment, AA is -C(O)-. 5 is alanine, cysteine, glycine, isoleucine, leucine, methionine, phenylalanine, valine, tryptophan, or tyrosine, and AA 6 is glycine. In one embodiment, AA 5 is leucine, and AA 6 is glycine. In one embodiment, AA 5 is valine, AA 6 is glycine. In one embodiment, AA 5 is phenylalanine, and AA 6 is glycine. In one embodiment, AA 5 is glycine and AA 6 is glycine. In one embodiment, AA 5 is not Balin.

[0095] In other embodiments, Z is H 2 N-A 3 -A.A. 4 -A.A. 5 -A.A. 6 In one embodiment, AA is -C(O)-. 5 is alanine, cysteine, glycine, isoleucine, leucine, methionine, phenylalanine, valine, tryptophan, or tyrosine, and AA 3 , A.A. 4 , or AA 6 Each of AA is glycine. 5 is leucine, and AA 3 , A.A. 4 , or AA6 Each of AA is glycine. 5 is valine, and AA 3 , A.A. 4 , or AA 6 Each of AA is glycine. 5 is phenylalanine, and AA 3 , A.A. 4 , or AA 6 Each of AA is glycine. 3 is glycine, AA 4 is phenylalanine, AA 5 is leucine, and AA 6 is glycine. In one embodiment, AA 3 , A.A. 4 , A.A. 5 and A.A. 6 Each of AA is glycine. 5 is not Balin.

[0096] In some embodiments, Z is hydrogen. In other embodiments, Z is H 2 N-AA 6 In one embodiment, AA is -C(O)-. 6 is glycine.

[0097] In some embodiments, Q is NR. In some embodiments, M is a bond. In some embodiments, J is a bond. In some embodiments, Y is NR.

[0098] In one embodiment, W is:

[0099] [ka]

[0100] [ka]

[0101] [ka]

[0102] [ka]

[0103] [ka]

[0104] and;

[0105] In the formula, R 2 is -OH or methoxy; and R 3 is hydrogen, -OH or methoxy.

[0106] In one embodiment, W is:

[0107] [ka]

[0108] [ka]

[0109] It is.

[0110] In one embodiment, W is:

[0111] [ka]

[0112] It is.

[0113] In one embodiment, -QXY- is:

[0114] [ka]

[0115] [ka]

[0116] and;

[0117] V is:

[0118] [ka]

[0119] or a bond; R 12 is hydrogen or Me; R 12 is R 14 together with R to form a piperidine ring; and R 11 is R 12 Together with

[0120] In one embodiment, Z is H 2 N-AA 5 -AA 6 -C(O)-;AA 5 is leucine, and AA 6 is glycine; QXY is:

[0121] [ka]

[0122] and W is:

[0123] [ka]

[0124] It is.

[0125] In one embodiment, Z is H 2 N-AA 5 -AA 6-C(O)- and AA 5 is valine, and AA 6 is glycine; QXY is:

[0126] [ka]

[0127] and W is:

[0128] [ka]

[0129] It is.

[0130] In one embodiment, Z is H 2 N-AA 5 -AA 6 -C(O)-;AA 5 is phenylalanine, and AA 6 is glycine; QXY is:

[0131] [ka]

[0132] and W is:

[0133] [ka]

[0134] It is.

[0135] In one embodiment, Z is H 2 N-AA 5 -AA 6 -C(O)-;AA 5 is glycine, and AA 6 is glycine; QXY is:

[0136] [ka]

[0137] and W is:

[0138] [ka]

[0139] It is.

[0140] In one embodiment, Z is H 2 N-AA 3 -AA 4 -AA 5 -AA 6 -C(O)-;AA 5 is leucine, and AA 3 , A.A. 4 , or AA 6 Each of is glycine; QXY is:

[0141] [ka]

[0142] and W is:

[0143] [ka]

[0144] It is.

[0145] In one embodiment, Z is H 2 N-AA 3 -AA 4 -AA 5 -AA 6 -C(O)-;AA 5 is valine, and AA 3 , A.A. 4、 AA 6 Each of is glycine; QXY is:

[0146] [ka]

[0147] and W is:

[0148] [ka]

[0149] It is.

[0150] In one embodiment, Z is H 2 N-AA 3 -AA 4 -AA 5 -AA 6 -C(O)-;AA 5 is phenylalanine, and AA 3 , A.A. 4、 AA 6 Each of is glycine; QXY is:

[0151] [ka]

[0152] and W is:

[0153] [ka]

[0154] It is.

[0155] In one embodiment, Z is H 2 N-AA 3 -AA 4 -AA 5 -AC(O)-;AA 3 is glycine and AA 4 is phenylalanine, and AA 5 is leucine, and AA 6is glycine; QXY is:

[0156] [ka]

[0157] and W is:

[0158] [ka]

[0159] It is.

[0160] In one embodiment, Z is H 2 N-AA 3 -AA 4 -AA 5 -AA 6 -C(O)-;AA 3 , A.A. 4 , A.A. 5 and A.A. 6 Each of is glycine; QXY is:

[0161] [ka]

[0162] and W is:

[0163] [ka]

[0164] It is.

[0165] In one embodiment, Z is H 2 N-AA 6 -C(O)-;AA 6 is glycine; QXY is:

[0166] [ka]

[0167] and W is

[0168] [ka]

[0169] It is.

[0170] In one embodiment, Z is hydrogen;

[0171] [ka]

[0172] and W is:

[0173] [ka]

[0174] It is.

[0175] In one embodiment, Z is H 2 N-AA 5 -AA 6 -C(O)-;AA 5 is leucine, and AA 6 is glycine; QXY is:

[0176] [ka]

[0177] and W is:

[0178] [ka]

[0179] It is.

[0180] In one embodiment, Z is H 2 N-AA 5 -AA 6 -C(O)-;AA 5 is valine, and AA 6 is glycine; QXY is:

[0181] [ka]

[0182] and W is:

[0183] [ka]

[0184] It is.

[0185] In one embodiment, Z is H 2 N-AA 5 -AA 6 -C(O)-;AA 5 is phenylalanine, and AA 6 is glycine; QXY is:

[0186] [ka]

[0187] and W is:

[0188] [ka]

[0189] It is.

[0190] In one embodiment, Z is H 2 N-AA 5 -AA 6-C(O)-;AA 5 is glycine, and AA 6 is glycine; QXY is:

[0191] [ka]

[0192] and W is:

[0193] [ka]

[0194] It is.

[0195] In one embodiment, Z is H 2 N-AA 3 -AA 4 -AA 5 -AA 6 -C(O)-;AA 5 is leucine, and AA 3 , A.A. 4 or A.A. 6 Each of is glycine; QXY is:

[0196] [ka]

[0197] and W is:

[0198] [ka]

[0199] It is.

[0200] In one embodiment, Z is H 2 N-AA 3 -AA 4 -AA 5 -AA 6-C(O)-;AA 5 is valine, and AA 3 , A.A. 4 , or AA 6 Each of is glycine; QXY is:

[0201] [ka]

[0202] and W is:

[0203] [ka]

[0204] It is.

[0205] In one embodiment, Z is H 2 N-AA 3 -AA 4 -AA 5 -AA 6 -C(O)-;AA 5 is phenylalanine, and AA 3 , A.A. 4 , or AA 6 Each of is glycine; QXY is:

[0206] [ka]

[0207] and W is:

[0208] [ka]

[0209] It is.

[0210] In one embodiment, Z is H 2 N-AA 3 -AA-AA5 -AA 6 -C(O)-;AA 3 is glycine, AA 4 is phenylalanine, AA 5 is leucine, and AA 6 is glycine; QXY is:

[0211] [ka]

[0212] and W is:

[0213] [ka]

[0214] It is.

[0215] In one embodiment, Z is H 2 N-AA 3 -AA 4 -AA 5 -AA 6 -C(O)-;AA 3 , A.A. 4 , A.A. 5 and A.A. 6 Each of is glycine; QXY is:

[0216] [ka]

[0217] and W is:

[0218] [ka]

[0219] It is.

[0220] In one embodiment, Z is H 2 N-AA6 -C(O)-;AA 6 is glycine; QXY is:

[0221] [ka]

[0222] and W is:

[0223] [ka]

[0224] It is.

[0225] In one embodiment, Z is hydrogen;

[0226] [ka]

[0227] and W is:

[0228] [ka]

[0229] It is.

[0230] In one embodiment, Z is H 2 N-AA 5 -AA 6 -C(O)-;AA 5 is leucine, and AA 6 is glycine; QXY is:

[0231] [ka]

[0232] and W is:

[0233] [ka]

[0234] It is.

[0235] In one embodiment, Z is H 2 N-AA 5 -AA 6 -C(O)-;AA 5 is valine, and AA 6 is glycine; QXY is:

[0236] [ka]

[0237] and W is:

[0238] [ka]

[0239] It is.

[0240] In one embodiment, Z is H 2 N-AA 5 -AA 6 -C(O)-;AA 5 is phenylalanine, and AA 6 is glycine; QXY is:

[0241] [ka]

[0242] and W is:

[0243] [ka]

[0244] It is.

[0245] In one embodiment, Z is H 2 N-AA 5 -AA 6 -C(O)-;AA 5 is glycine, and AA 6 is glycine; QXY is:

[0246] [ka]

[0247] and W is:

[0248] [ka]

[0249] It is.

[0250] In one embodiment, Z is H 2 N-AA 3 -AA 4 -AA 5 -AA 6 -C(O)-;AA 5 is leucine, and AA 3 , A.A. 4 , or AA 6 Each of is glycine; QXY is:

[0251] [ka]

[0252] and W is:

[0253] [ka]

[0254] It is.

[0255] In one embodiment, Z is H 2 N-AA 3 -AA 4 -AA 5 -AA 6 -C(O)-;AA 5 is valine, and AA 3 , A.A. 4 , or AA 6 Each of is glycine; QXY is:

[0256] [ka]

[0257] and W is:

[0258] [ka]

[0259] It is.

[0260] In one embodiment, Z is H 2 N-AA 3 -AA 4 -AA 5 -AA 6 -C(O)-;AA 5 is phenylalanine, and AA 3 , A.A. 4 or A.A. 6 Each of is glycine; QXY is:

[0261] [ka]

[0262] and W is:

[0263] [ka]

[0264] It is.

[0265] In one embodiment, Z is H 2 N-AA 3 -AA 4 -AA 5 -AA 6 -C(O)-;AA 3 is glycine and AA 4 is phenylalanine, and AA 5 is leucine, and AA 6 is glycine; QXY is:

[0266] [ka]

[0267] and W is:

[0268] [ka]

[0269] It is.

[0270] In one embodiment, Z is H 2 N-AA 3 -AA 4 -AA 5 -AA 6 -C(O)-;AA 3 , A.A. 4 , A.A. 5 and A.A. 6 Each of is glycine; QXY is:

[0271] [ka]

[0272] and W is:

[0273] [ka]

[0274] is.

[0275] In one embodiment, Z is H 2 N-AA 6 -C(O)-; AA 6 is glycine; Q-X-Y is:

[0276]

Chem.

[0277] and W is:

[0278]

Chem.

[0279] is.

[0280] In one embodiment, Z is hydrogen; Q-X-Y is:

[0281]

Chem.

[0282] and W is:

[0283]

Chem.

[0284] is.

[0285] Other active moieties that can be modified for use in the complexes of the present invention include the following structures:

[0286]

Chem.

[0287] In some embodiments, the active moiety is an anti-obesity compound. In other embodiments, the active moiety is a molecule that inhibits methionine aminopeptidase-2 (MetAP2), such as fumagillin, fumagillol, or its analogs, derivatives, salts or esters. Furthermore, preferred MetAP2 inhibitors are described in U.S. Patent No. 6,242,494 to Craig et al., U.S. Patent No. 6,063,812 to Hong et al., U.S. Patent No. 6,887,863 to Craig et al., U.S. Patent No. 7,030,262 to BaMaung et al., and U.S. Patent No. 7,491,718 to Comess et al., each of which is incorporated by reference in its entirety. Additional preferred MetAP2 inhibitors include those described by Wang et al., “Correlation of tumor growth suppression and methionine aminopeptidase-2 activity blockade using an orally active inhibitor,” PNAS 105(6)1838-1843 (2008); Lee et al., “Design, Synthesis, and Antiangiogenic Effects of a Series of Potent Novel Fumagillin Analogues,” Chem. Pharm. Bull. 55(7)1024-1029 (2007); Jeong et al., “Total synthesis and antioxidant activity of cyclopentane analogues of fumagillol,” Bioorganic and Medicinal Chemistry Letters 15,3580-3583 (2005); and Arico-Muendel et al., “Carbamate Analogues of Fumagillin as Potent, Targeted Inhibitors of Methionine Aminopeptidase-2," J. Med. Chem. 52, 8047-8056 (2009); and WO 2010 / 003475 to Heinrich et al.

[0288] Fumagillin is a small molecule used as an antibacterial and antiprotozoal agent. Its physicochemical properties and methods of preparation are known (see U.S. Pat. No. 2,803,586 and Turner, JR et al., The Stereochemistry of Fumagillin, Proc. Natl. Acad. Sci. 48, 733-735 (1962)). The fermentation product, fumagillin, is hydrolyzed to give the alcohol fumagillol, which may be converted to various derivatives, including carbamoyl fumagillol, MW325. The synthesis and preparation of carbamoyl fumagillol and several small molecule derivatives are described in U.S. Pat. No. 5,166,172.

[0289] Fumagillin and related compounds are believed to exert their biological effects through inhibition of MetAP2, an enzyme that removes N-terminal methionine from nascent cellular proteins (see Tucker, LA et al., "Ectopic Expression of Methionine Aminopeptidase-2 Causes Cell Transformation and Stimulates Proliferation," Oncogene 27, 3967 (2008)).

[0290] Carbamoyl fumagillol and derivatives, as well as other MetAP2 inhibitors, have shown therapeutic efficacy in preclinical and clinical studies. These compounds inhibit cell proliferation and angiogenesis, as described in U.S. Pat. No. 5,166,172. Fumagillin analogs or derivatives, such as CKD-732 and PI-2458, have been well studied in various systems, as described in Bernier et al., "Fumagillin class inhibitors of methionine aminopeptidase-2," Drugs of the Future 30(5):497-508, 2005.

[0291] The anti-obesity effects of fumagillin and its analogs are known. Rupnick et al., "Adipose tissue mass can be regulated through the vasculature," PNAS 99, 10730-10735, 2002, describes weight loss in ob / ob mice with TNP-470 at daily doses ranging from 2.5 mg / kg to 10 mg / kg. Brakenhielm describes the prevention of obesity with TNP-470 at doses of 15 or 20 mg / kg every other day: "The Angiogenesis Inhibitor, TNP-470, Prevents Diet-Induced and Genetic Obesity in Mice," Circulation Research 94:1579-1588, 2004. Kim et al., "Assessment of the anti-obesity effects of the TNP-470 analog, CKD-732," J Molecular Endocrinology 38, 455-465, 2007, describe weight loss in C57BL / 6J mice and SD rats at a dose of 5 mg / kg / day. Lijnen et al., "Fumagillin reduces adipose tissue formation in murine models of nutritionally induced obesity," Obesity 12, 2241-2246, 2010, describe that daily oral delivery of 1 mg / kg fumagillin results in weight loss in C57BL / 6 mice.

[0292] One of these derivatives, chloroacetylcarbamoyl fumagillol (TNP-470), has been studied extensively. (See H. Mann-Steinberg et al., "TNP-470: The Resurrection of the First Synthetic Angiogenesis Inhibitor", Chapter 35 in Folkman and Figg, Angiogenesis: An Integrative Approach from Science to Medicine, Springer NY (2008)). TNP-470 has shown activity against many cancers, including lung, cervical, ovarian, breast and colon cancers. Due to dose-limiting neurotoxicity, TNP-470 has been tested using multiple dosing regimens, but these attempts to limit its toxicity have been unsuccessful. Thus, TNP-470 has proven too toxic for use in humans. TNP-470 has a short half-life and requires prolonged intravenous administration for therapeutic use. Carbamoyl fumagillol, a metabolite of TNP-470, has a half-life of 12 minutes in humans (see Herbst et al., "Safety and Pharmacokinetic Effects of TNP-470, an Angiogenesis Inhibitor, Combined with Paclitaxel in Patients with Solid Tumors: Evidence for Activity in Non-Small-Cell Lung Cancer," Journal of Clinical Oncology 20(22)4440-4447 (2002). Furthermore, fumagillin and its derivatives are hydrophobic and difficult to formulate.

[0293] Despite the known usefulness of fumagillin derivatives, their use in therapy has not been successful due to the inability to overcome the problems of low water solubility, short half-life value, and neurotoxic side effects of these compounds.Based on the previously observed dose-limiting neurotoxicity, the MTD of TNP-470 in combination with paclitaxel has been determined to be 60 mg / m2 administered three times a week: Herbst et al., "Safety and pharmacokinetic effects of TNP-470, an angiogenesis inhibitor, combined with paclitaxel in patients with solid tumors: evidence for activity in non-small-cell lung Cancer", Journal of Clinical Oncology 20, 4440-4447, 2002. Similarly, Shin et al., "A Phase 1 pharmacokinetic and pharmacodynamics study of CKD-732, an antiangiogenic agent, in patients with refractory solid cancer"Investigational New Drugs 28, 650-658, 2010" reported that the MTD of CKD-732 is 15 mg / m2 / day, administered on a 4-day schedule due to confusion and insomnia. Thus, the compounds of the present invention are more potent, exhibit reduced toxicity (lower neurotoxicity), have improved water solubility, are more stable, and / or have a longer half-life (serum half-life) than currently known fumagillin derivatives.

[0294] The term "reduced toxicity" as used herein has its ordinary meaning as understood by one of ordinary skill in the art. By way of example only and not intended to limit the meaning of the term in any way, administration of a fumagillin analog conjugate results in fewer side effects in an open field test in mice compared to administration of the fumagillin analog alone.

[0295] The phrase "improved water solubility" has its ordinary meaning as understood by those of skill in the art. By way of example only and not intended to limit the meaning of the term in any way, the following description of the term is instructive: the fumagillin analog becomes more soluble in water by being covalently incorporated into the complex than the unbound fumagillin analog would be soluble in water alone.

[0296] The phrase "longer half-life" has its ordinary meaning as understood by one of ordinary skill in the art. By way of example only and not intended to limit the meaning of the term in any way, the following description of the term is instructive: any appreciable increase in the length of time required to deactivate a fumagillin conjugate, either in vivo or in vitro, as compared to the half-life of the fumagillin analog alone, either in vivo or in vitro.

[0297] Without being bound by any theory, the nonenzymatic action of MetAP2, which inhibits the activity of extracellular signal-regulated kinases 1 and 2 (ERK1 / 2), may be important, as may the binding of the eukaryotic translation initiation factor, eIF, by MetAP2. Cellular responses to MetAP2 inhibition, reflecting potential ERK-related processes, may include suppression of sterol regulatory element-binding protein (SREBP) activity, leading to reduced lipid and cholesterol biosynthesis. Interestingly, changes in the expression patterns of liver and adipose tissue genes after long-term (approximately 9 months) fumagillin exposure suggest that inhibition of MetAP2 may alter the relative abundance of factors involved in inflammation, consistent with a reduction in ERK-dependent cellular processes. The putative mechanism of MetAP2 inhibition, leading to the mobilization of stored fat as an energy source by the body and the catabolism of free fatty acids, is supported by the changes in plasma β-hydroxybutyrate, adiponectin, leptin, and FGF21 observed in previous studies. The increase in the levels of adiponectin and FGF21, the major catabolic hormones, accompanied by the appearance of ketone bodies (β-hydroxybutyrate), suggests that the inhibition of MetAP2 by the compound of the present invention, conjugated or modified fumagillin, fumagillol, or its analogs, derivatives, salts or esters, promotes energy expenditure, fat utilization, and lipid excretion. The reduction in leptin observed in previous studies and the studies provided herein is also consistent with the reduction in total adipose tissue and negative energy balance. It is also possible that the compound of the present invention, conjugated or modified fumagillin, fumagillol, or its analogs, derivatives, salts or esters, forms a covalent bond with MetAP2, thereby irreversibly inhibiting and shutting down existing enzymes until a pool of newly produced MetAP2 is generated in target tissues (e.g., liver and adipose tissue).

[0298] In certain embodiments, the conjugated or modified fumagillin, fumagillol, or analogs, derivatives, salts, or esters thereof of the compounds of the invention have the structure of the following formula, for example, as shown in Table 1:

[0299] [Table 1A]

[0300] [Table 1B]

[0301] [Table 1C]

[0302] [Table 1D]

[0303] [Table 1E]

[0304] [Table 1F]

[0305] [Table 1G]

[0306] * wherein the polymer is:

[0307] [ka]

[0308] and preferably has the structure:

[0309] [ka]

[0310] It has the structure:

[0311] For purposes of this invention, the chemical elements are identified according to the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 67th Ed., 1986-87, inside cover.

[0312] The term "alkyl" refers to a fully saturated, branched or unbranched carbon chain radical having the specified number of carbon atoms, or, if not specified, up to 30 carbon atoms. For example, "lower alkyl" refers to alkyls having 1 to 10 carbon atoms, such as methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, and octyl, as well as alkyl positional isomers thereof. Alkyl having 10 to 30 carbon atoms includes decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, octadecyl, nonadecyl, eicosyl, heneicosyl, docosyl, tricosyl, and tetracosyl. In certain embodiments, a straight or branched chain alkyl has 30 or fewer carbon atoms in its backbone (e.g., C for straight chain). 1 -C 30 , C for branched chains 3 -C 30 ), and more preferably have 20 or fewer carbon atoms. Likewise, certain cycloalkyls have from 3-10 carbon atoms in their ring structure, and can have 5, 6 or 7 carbons in the ring structure.

[0313] Unless the number of carbon atoms is otherwise specified, "lower alkyl" as used herein refers to an alkyl group as defined above, but having 1-10 carbon atoms, or 1-6 carbon atoms in its main chain structure, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, and tert-butyl. Similarly, "lower alkenyl" and "lower alkynyl" have similar chain lengths. Throughout this specification, certain alkyl groups are lower alkyls. In certain embodiments, a substituent designated herein as alkyl is a lower alkyl.

[0314] The term "carbocycle" as used herein refers to an aromatic or non-aromatic ring in which each atom of the ring is carbon.

[0315] The term "aryl" as used herein includes 5-, 6-, and 7-membered monocyclic aromatic groups that may contain 0-4 heteroatoms, such as benzene, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, triazole, pyrazole, pyridine, pyrazine, pyridazine, pyrimidine, and the like. Aryl groups having heteroatoms in the ring structure may also be referred to as "aryl heterocycles" or "heteroaromatics." The aromatic ring may contain, at one or more ring positions, substituents as described above, such as halogen, azido, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, alkoxyl, amino, nitro, sulfhydryl, imino, amide, phosphonate, phosphinate, carbonyl, carboxyl, silyl, ether, alkylthio, sulfonyl, sulfonamide, ketone, aldehyde, ester, heterocyclyl, aromatic or heteroaromatic moiety, -CF 3 , -CN, or the like. The term "aryl" also includes polycyclic ring structures having two or more rings in which two or more carbons are shared between two adjacent rings (the rings are "fused rings"), where at least one ring is aromatic and the other rings may be, for example, cycloalkyl, cycloalkenyl, cycloalkynyl, aryl and / or heterocyclyl.

[0316] "Alkenyl" refers to any branched or unbranched unsaturated carbon chain radical having a specific number of carbon atoms, or up to 26 carbon atoms if no limit on the number of carbon atoms is specified; and having one or more double bonds in the radical. Alkenyls having 6 to 26 carbon atoms are exemplified by the various isomeric forms of hexenyl, heptenyl, octenyl, nonenyl, decenyl, undecenyl, dodenyl, tridecenyl, tetradecenyl, pentadecenyl, hexadecenyl, heptadecenyl, octadecenyl, nonadecenyl, eicosenyl, heneicosoenyl, docosenyl, tricosenyl, and tetracosenyl, where the unsaturated bond may be located anywhere in the radical and the double bond may have either the (Z) or (E) configuration.

[0317] The term "alkynyl" refers to a hydrocarbyl radical within the scope of alkenyl, but having one or more triple bonds.

[0318] The term "alkoxyl" or "alkoxy" as used herein refers to an alkyl group, as defined below, having an oxygen radical attached thereto. Representative alkoxy groups include methoxy, ethoxy, propoxy, tert-butoxy, and the like. An "ether" is two hydrocarbons covalently linked by an oxygen. Thus, substituents of an alkyl that make it an ether are -O-alkyl, -O-alkenyl, -O-alkynyl, -O-(CH 2 ) m -R 1 (m and R 1 is or resembles an alkoxyl, such as may be represented by one of the following formulas:

[0319] The terms "heterocyclyl" or "heterocyclic group" refer to 3- to 10-membered ring structures, more preferably 3- to 7-membered rings, whose ring structures include one to four heteroatoms. The heterocycle may be polycycle. Heterocyclyl groups include, for example, thiophene, thianthrene, furan, pyran, isobenzofuran, chromene, xanthene, phenoxathiin, pyrrole, imidazole, pyrazole, isothiazole, isoxazole, pyridine, pyrazine, pyrimidine, pyridazine, indolizine, isoindole, indole, indazole, purine, quinolizine, isoquinoline, quinoline, phthalazine, naphthyridine, quinoxaline, quinazoline, cinnoline, pteridine, carbazole, carboline, phenanthridine, acridine, pyrimidine, phenanthroline, phenazine, phenarsazine, phenothiazine, furazan, phenoxazine, pyrrolidine, oxolane, thiolane, oxazole, piperidine, piperazine, morpholine, lactones, lactams such as azetidinones and pyrrolidinones, sultams, sultones, and the like. The heterocycle may contain, at one or more positions, the above-mentioned substituents, such as, for example, halogen, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, amino, nitro, sulfhydryl, imino, amido, phosphate, phosphonate, phosphinate, carbonyl, carboxyl, silyl, sulfamoyl, sulfamyl, ether, alkylthio, sulfonyl, ketone, aldehyde, ester, heterocyclyl, aromatic or heteroaromatic moiety, -CF 3 , -CN, or the like.

[0320] The term "alkylthio" refers to an alkyl group, as defined above, having a sulfur radical attached thereto. In certain embodiments, the "alkylthio" moiety includes -(S)-alkyl, -(S)-alkenyl, -(S)-alkynyl, and -(S)-(CH 2 ) m -R 1 (m and R 1 (wherein R is an integer from 1 to 5; R is an integer from 1 to 5; and R is an integer from 1 to 5). Representative alkylthio groups include methylthio, ethylthio, and the like.

[0321] As used herein, the term "nitro" means -N 2 the term "halogen" refers to F, Cl, Br, or I; the term "sulfhydryl" means -SH; the term "hydroxyl" means -OH; and the term "sulfonyl" means -SO 2 - means.

[0322] The terms "amine" and "amino" are art-recognized and refer to both unsubstituted and substituted amines, such as those of the general formula:

[0323] [ka]

[0324] refers to a moiety that can be represented by

[0325] In the formula, R 3 , R 5 and R 6 are each independently hydrogen, alkyl, alkenyl, -(CH 2 ) m -R 1 or R 3 and R 5 together with the N atom to which they are attached form a heterocycle having 4 to 8 atoms in the ring structure; R 1 represents alkenyl, aryl, cycloalkyl, cycloalkenyl, heterocyclyl, or polycyclyl; and m is 0 or an integer ranging from 1 to 8. In certain embodiments, R 3 or R 5 Only one of R may be a carbonyl, e.g., R 3 , R 5 and the nitrogen together do not form an imide. 3 and R 5 (and optionally R 6 ) are each independently hydrogen, an alkyl, an alkenyl, or -(CH 2 ) m -R1 Thus, the term "alkylamine" as used herein refers to an amine group, as defined above, to which is attached a substituted or unsubstituted alkyl, i.e., at least one R 3 and R 5 is an alkyl group. In certain embodiments, the amino group or alkylamine is basic, i.e., the pK a pK ≧7.00 a The protonated forms of these functional groups have a pK with respect to water higher than 7.00. a has.

[0326] The term "carbonyl" (C(O)) is art-recognized and has the general formula:

[0327] [ka]

[0328] and a portion that can be represented by

[0329] In the formula, X represents a bond, or an oxygen or sulfur atom, and R 7 is hydrogen, alkyl, alkenyl, -(CH 2 ) m -R 1 or a pharma- ceutically acceptable salt thereof; 8 is hydrogen, alkyl, alkenyl or -(CH 2 ) m -R 1 (m and R 1 X is oxygen and R 7 or R 8 Where X is oxygen and R is not hydrogen, the formula represents an "ester". 7 is as defined above, the moiety is referred to herein as a carboxyl group, and in particular R 7 Where X is oxygen and R 8Where R is hydrogen, the formula represents a "formate." In general, where the oxygen atom of the above formula is replaced with a sulfur, the formula represents a "thiocarbonyl" group. 7 or R 8 Where X is not hydrogen, the above formula represents a "thioester" group. 7 Where X is a sulfur and R is a hydrogen, the above formula represents a "thiocarboxylic acid" group. 8 Where X is hydrogen, the above formula represents a "thioformate" group. 7 Where X is a bond and R is not hydrogen, the above formula represents a "ketone" group. 7 Where is hydrogen, the above formula represents an "aldehyde" group.

[0330] As used herein, the term "substituted" is intended to include all permissible substituents of organic compounds. In a broad aspect, the permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and nonaromatic substituents of organic compounds. Exemplary substituents include, for example, those described above. The permissible substituents can be one or more and can be the same or different in appropriate organic compounds. For purposes of this invention, heteroatoms such as nitrogen can have hydrogen substituents and / or any permissible substituents of organic compounds described herein that satisfy the valence of the heteroatom. This invention is not intended to be limited in any manner by the permissible substituents of organic compounds. It is understood that the phrases "substituted" or "substituted with" include the implicit proviso that such substitution is based on the permissible valences of the substituted atom and substituent, and that the substitution results in a stable compound, e.g., a compound that does not spontaneously undergo transformation by rearrangement, cyclization, elimination, and the like.

[0331] The term "sulfamoyl" is art-recognized and refers to a group represented by the general formula:

[0332] [ka]

[0333] and a portion that can be represented by

[0334] In the formula, R 3 and R 5 is as defined above.

[0335] The term "sulfate" is art-recognized and has the general formula:

[0336] [ka]

[0337] and a portion that can be represented by

[0338] In the formula, R 7 is as defined above.

[0339] The term "sulfamide" is art-recognized and has the general formula:

[0340] [ka]

[0341] and a portion that can be represented by

[0342] In the formula, R 2 and R 4 is as defined above.

[0343] The term "sulfonate" is art-recognized and has the general formula:

[0344] [ka]

[0345] and a portion that can be represented by

[0346] In the formula, R 7 is an electron pair, hydrogen, alkyl, cycloalkyl, or aryl.

[0347] The term "sulfoxide" or "sulfinyl" as used herein refers to a group having the general formula:

[0348] [ka]

[0349] "" refers to a portion that can be represented by

[0350] In the formula, R 12 is selected from the group consisting of hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aralkyl, or aryl.

[0351] Analogous substitutions on alkenyl and alkynyl groups can form, for example, aminoalkenyls, aminoalkynyls, amidoalkenyls, amidoalkynyls, iminoalkenyls, iminoalkynyls, thioalkenyls, thioalkynyls, carbonyl-substituted alkenyls or alkynyls.

[0352] As used herein, the definition of each expression, e.g., alkyl, m, n, etc., when it occurs more than one time in any structure, is intended to be independent of its definition elsewhere in the same structure.

[0353] The term "amino acid" is intended to encompass all compounds, whether natural or synthetic, that contain both amino and acid functional groups, including amino acid analogs and derivatives. In certain embodiments, the amino acids contemplated in the present invention are naturally occurring amino acids present in proteins, or naturally occurring anabolic or catabolic products of such amino acids that contain amino and carboxyl groups. Naturally occurring amino acids are identified throughout the present specification by the conventional three-letter and / or one-letter abbreviations that correspond to the common names of the amino acids according to the following table. The abbreviations are accepted in the field of peptides and are recommended by the IUPAC-IUB Commission on Biochemical Nomenclature.

[0354] The term "amino acid residue" refers to an amino acid. In general, the abbreviations used herein to refer to naturally occurring amino acids are based on the recommendations of the IUPAC-IUB Commission on Biochemical Terminology (Biochemistry (1972) 11:1726-1732). For example, Met, He, Leu, Ala, and Gly represent the "residues" of methionine, isoleucine, leucine, alanine, and glycine, respectively. Residue refers to the radical obtained by removing the OH moiety of the carboxyl group and the hydrogen moiety of the α-amino group from the corresponding α-amino acid.

[0355] The term "amino acid side chain" refers to that portion of an amino acid residue excluding the main chain, as defined in KD Kopple, "Peptides and Amino Acids," WA Benjamin Inc., New York and Amsterdam, 1966, pages 2 and 33; examples of such side chains of common amino acids are -CH 2 CH 2 SCH 3 (methionine side chain), -CH 2 (CH 3 )-CH 2 CH 3 (side chain of isoleucine), -CH 2 CH(CH 3 ) 2(the side chain of leucine) or H- (the side chain of glycine). These side chains are pendant from the main chain Cα carbon.

[0356] The term "peptide" as used herein refers to a sequence of amino acid residues linked by peptide bonds or modified peptide bonds. The term "peptide" is intended to encompass peptide analogs, peptide derivatives, peptidomimetics, and peptide variants. The term "peptide" is understood to include peptides of any length. Peptide sequences described herein are written according to generally accepted convention, with the N-terminal amino acid on the left and the C-terminal amino acid on the right (e.g., H 2 N-AA 1 -AA 2 -AA 3 -AA 4 -AA 5 -AA 6 -CO 2 H).

[0357] Certain compounds of the present invention may exist in specific geometric or stereoisomeric forms. The present invention contemplates all such compounds, including cis and trans isomers, R- and S-enantiomers, diastereomers, (D)-isomers, (L)-isomers, racemic mixtures thereof, and other mixtures thereof, to be within the scope of the present invention. Additional asymmetric carbon atoms may be present in a substituent, such as an alkyl group. All such isomers, and mixtures thereof, are intended to be included in the present invention. Any recitation of a specific isomer is merely exemplary (e.g., recitation of a trans isomer also includes the cis isomer).

[0358] For example, if a particular enantiomer of a compound of the invention is desired, it can be prepared by asymmetric synthesis or derivatization with a chiral auxiliary, in which case the resulting mixture of diastereomers can be separated and the pure desired enantiomer obtained by cleavage of the auxiliary. Alternatively, if the molecule contains a basic functional group, such as an amino group, or an acidic functional group, such as a carboxyl group, the salt of the diastereomer can be formed with a suitable optically active acid or base, and the diastereomers so formed can then be resolved by fractional crystallization or chromatographic means known in the art, followed by recovery of the pure enantiomer.

[0359] Synthesis of Compounds of the Invention The synthetic methods of the present invention tolerate a wide range of functional groups; therefore, various substituted starting materials can be used. The methods generally provide the desired final compound at or near the end of the overall process, although in some cases it may be desirable to further convert the compound to its pharma- ceutically acceptable salt, ester, or prodrug.

[0360] The compounds of the present invention can be prepared in a variety of ways using standard synthetic methods and procedures known to those skilled in the art or that will be apparent to those skilled in the art in light of the teachings herein, using commercially available starting materials, compounds known in the literature, or ready-to-use intermediates. Standard synthetic methods and procedures for the preparation of organic molecules and functional group transformations and manipulations can be obtained from the relevant scientific literature or standard textbooks in the field. Without being limited to any one or several sources, classical references, such as Smith, MB, March, J., March's Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, 5, pp. 111-115, 2002, which are incorporated herein by reference, may be used. thedition, John Wiley & Sons: New York, 2001; and Greene, TW, Wuts, PGM, Protective Groups in Organic Synthesis, 3 rd edition, John Wiley & Sons: New York, 1999, is a useful and recognized reference in organic synthesis known to those skilled in the art. The following descriptions of synthetic methods are intended to illustrate, but not limit, basic procedures for the preparation of the compounds of the present invention.

[0361] The compounds of the present invention can be conveniently prepared by various methods known to those skilled in the art.The compounds of the present invention can be prepared according to the schemes and examples provided herein from commercially available starting materials or starting materials that can be prepared using procedures described in the literature.The compounds of the present invention and their synthesis are further described in WO 2011 / 150088 and WO 2011 / 150022.Each of these publications is incorporated by reference in its entirety for all purposes.

[0362] Pharmaceutical Compositions The present invention also provides pharmaceutical compositions comprising a compound of the present invention, or a pharma- ceutically acceptable salt, solvate, diastereomer, or polymorph thereof, and a pharma- ceutically acceptable carrier or excipient.

[0363] As used herein, "pharmaceutical acceptable excipient" or "pharmaceutical acceptable carrier" is intended to include any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like, compatible with pharmaceutical administration. Suitable carriers are described in the latest edition of Remington's Pharmaceutical Sciences, a standard reference text in the field. Preferred examples of such carriers or diluents include, but are not limited to, water, saline, Ringer's solution, dextrose solution, and 5% human serum albumin.

[0364] Pharmaceutically acceptable carriers include solid carriers such as lactose, terra alba, sucrose, talc, gelatin, agar, pectin, acacia, magnesium stearate, stearic acid, and the like. Preferred liquid carriers include syrup, peanut oil, olive oil, water, and the like. Similarly, carriers or diluents can include delay materials known in the art, such as glyceryl monostearate or glyceryl distearate, alone or with wax, ethylcellulose, hydroxypropylmethylcellulose, methyl methacrylate, or the like. Other fillers, excipients, flavorings, and other additives, such as those known in the art, can also be included in the pharmaceutical compositions of the present invention. Liposomes and non-aqueous excipients, such as fixed oils, can also be used. The use of such media and agents for pharma-ceutically active substances is known in the art. Except insofar as any conventional media or agent is incompatible with the active compound, its use in the composition is contemplated. Supplementary active compounds can also be incorporated into the compositions. In certain embodiments, the pharmaceutical composition comprises DMSO.

[0365] The term "pharmaceutical acceptable salts" refers to relatively non-toxic, inorganic and organic acid addition salts of compounds. These salts can be prepared during the final isolation and purification of the compounds, or in situ by separately reacting the purified compounds in free base form with a suitable organic or inorganic acid and isolating the salt thus formed. Representative salts include hydrobromide, hydrochloride, sulfate, bisulfate, phosphate, nitrate, acetate, valerate, oleate, palmitate, stearate, laurate, benzoate, lactate, phosphate, tosylate, citrate, maleate, fumarate, succinate, tartrate, naphthylate, mesylate, glucoheptonate, lactobionate, and laurylsulfonate, and the like. Representative alkali or alkaline earth salts include lithium, sodium, potassium, calcium, magnesium, and aluminum salts, and the like. Representative organic amines useful for the formation of base addition salts include ethylamine, diethylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine, and the like (see, for example, Berge et al. (1977) "Pharmaceutical Salts", J. Pharm. Sci. 66:1-19).

[0366] The phrase "pharmacologically acceptable" is used herein to refer to ligands, materials, compositions, and / or dosage forms that are substantially non-pyrogenic and suitable for use in contact with the tissues of human beings and animals, within the scope of sound medical judgment, without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0367] As used herein, the term "metabolite" means a metabolite of a compound of the present invention, or a pharma- ceutically acceptable salt, solvate, diastereomer, or polymorph thereof, that exhibits activity in vivo similar to that of the compound of the present invention, or a pharma- ceutically acceptable salt, solvate, diastereomer, or polymorph thereof.

[0368] As used herein, the term "prodrug" refers to a compound of the invention, or a pharma- ceutically acceptable salt, solvate, diastereomer, and polymorph thereof, covalently linked to one or more pro-moieties, such as, for example, an amino acid moiety or other water-soluble moiety. The compound of the invention, or a pharma- ceutically acceptable salt, solvate, diastereomer, and polymorph thereof, can be released from the pro-moiety via hydrolytic, oxidative, and / or enzymatic release mechanisms. In one embodiment, the pro-drug composition of the invention exhibits the added benefits of increased water solubility, improved stability, and improved pharmacokinetic profile. The pro-moiety can be selected to obtain the desired pro-drug properties. For example, the pro-moiety, e.g., an amino acid moiety such as a phosphate in R4 or other water-soluble moiety, can be selected based on solubility, stability, bioavailability, and / or in vivo delivery or uptake. Examples of prodrugs include, but are not limited to, esters (e.g., acetate, dialkylaminoacetate, formate, phosphate, sulfate and benzoate derivatives) and carbamates (e.g., N,N-dimethylaminocarbonyl) of hydroxy functional groups, esters (e.g., ethyl ester, morpholinoethanol ester) of carboxyl functional groups, N-acyl derivatives (e.g., N-acetyl), N-Mannich bases, Schiff bases and enaminones of amino functional groups, and oximes, acetals, ketals, enol esters and the like of ketone and aldehyde functional groups in the compounds of the present invention; see Bundegaard, H., Design of Prodrugs, pi -92, Elesevier, New York-Oxford (1985).

[0369] Treatment method The present invention provides a method for inducing or causing weight loss in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of at least one compound of the present invention to induce or cause weight loss.In some embodiments, the subject is overweight or obese.In some embodiments, inducing or causing weight loss is increasing weight loss.

[0370] The present invention also provides a method of preventing or slowing weight gain in a subject at risk thereof, comprising administering to the subject a therapeutically effective amount of at least one compound of the present invention to prevent or slow weight gain, in certain embodiments, the subject is at risk of becoming overweight or obese.

[0371] The present invention provides methods of treating obesity in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of at least one compound of the present invention to treat or ameliorate obesity.

[0372] The present invention also provides methods for preventing or delaying the onset of obesity in a subject at risk thereof, comprising administering to the subject a therapeutically effective amount of at least one compound of the present invention to prevent or delay the onset of obesity.

[0373] The present invention provides methods of treating metabolic syndrome or one or more components thereof in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of at least one compound of the present invention to treat or ameliorate metabolic syndrome or one or more components thereof.

[0374] The present invention also provides a method for preventing or delaying the onset of metabolic syndrome or one or more of its components in a subject at risk thereof, comprising administering to the subject a therapeutically effective amount of at least one compound of the present invention to prevent or delay the onset of metabolic syndrome or one or more of its components.

[0375] The present invention also provides a method for reducing body weight in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of at least one compound of the present invention to reduce body weight. In some embodiments, the subject is overweight or obese. In some embodiments, the subject is in need of reducing excess adipose tissue.

[0376] Obesity and being overweight refer to a subject having an excess of fat relative to their lean body mass. Excessive fat accumulation is associated with an increase in the size (hypertrophy or adiposity) and number (hyperplasia) of adipose tissue cells. Obesity can be due to a variety of factors, whether genetic (e.g. Prader-Willi syndrome) or environmental. Obesity is measured in various ways in terms of absolute body weight, weight-to-weight ratio, degree of excess body fat, distribution of visceral or subcutaneous fat, and social and aesthetic norms. A common measure of body fat is the Body Mass Index (BMI). BMI represents the ratio of body weight (expressed in kilograms) to the square of height (expressed in meters). Body Mass Index can be calculated accurately using the following formula: SI units: BMI = weight (kg) / (height) 2 (m 2 ), or in US units: BMI = (weight (lbs) * 703) / (height 2 (inch 2 ).

[0377] As used herein, "overweight" is defined as a weight gain or weight exceeding 25 kg / m2 in an otherwise healthy adult. 2 ~29.9kg / m 2 As used herein, "obesity" refers to a condition in which an otherwise healthy adult has a BMI of 30 kg / m 2Obesity is defined as a condition in which the body mass index (BMI) is greater than or equal to 35 kg / m2. 2 Adults with a BMI above 40 to 44.9 kg / m are considered to be "severely obese" or "severely obese." 2 Adults with a BMI of 35 kg / m 2 Adults with a BMI above 45 kg / m2 and who have at least one obesity-related health condition are referred to as "morbidly obese" or "morbidly obese." 2 Adults with a BMI above this level are termed "super obese" or "super obese." In children, the definition of overweight and obesity takes into account the effects of age and sex on body fat.

[0378] Different countries may define obesity and overweight by different BMIs. The term "obesity" is intended to encompass definitions in all countries. For example, in Asians, the increased risk associated with obesity occurs at lower body mass index (BMI). In Asian countries, including Japan, "obesity" is defined as a condition in which a subject with at least one obesity-induced or obesity-related comorbidity that requires or would be improved by weight loss has a BMI of 25.0 kg / m2 or more. 2 It refers to the condition of having a BMI above 10. South and Central Americans tend to be classified as more similar to Asians than Europeans or North Americans.

[0379] BMI cannot account for the fact that excess adipose tissue occurs selectively in different parts of the body, and the development of adipose tissue may be more dangerous in some parts of the body than in other parts of the body. For example, "central obesity", which is typically associated with an "apple-shaped" body type, is caused by excess fat accumulation, especially in the abdominal region, including abdominal fat and visceral fat, and has a higher risk of comorbidities than "peripheral obesity", which is typically associated with a "pear-shaped" body type, and is caused by excess fat accumulation, especially in the waist. Measurement of waist / hip ratio (WHR) can be used as an index of central obesity. The minimum WHR indicating central obesity has been set in various ways, and adults with central obesity typically have a WHR of about 0.85 or more for women and about 0.9 or more for men.

[0380] Disease assessment is performed by standard methods known in the art, such as by monitoring appropriate markers. For example, for obesity, the following markers may be monitored: weight, BMI, body composition survey, body fat distribution, central fat distribution, food or calorie intake, behavioral measures of hunger and satiety, metabolic rate, and obesity-related comorbidities.

[0381] A method for determining whether a subject is overweight or obese, which describes the ratio of excess adipose tissue to lean body mass, includes obtaining information of the subject's body composition. Body composition can be obtained by measuring the thickness of subcutaneous fat at multiple locations on the body, such as the abdomen, subscapular region, arms, buttocks, and thighs. These measurements are then used to estimate total body fat with a margin of error of about 4 percentage points. Another method is bioelectrical impedance analysis (BIA), which uses the resistance of an electric current passing through the body to estimate body fat. Another method uses a large tank of water to measure body buoyancy. More body fat results in more buoyancy and more muscle mass results in a tendency to sink. Another method is fan beam dual energy x-ray absorptiometry (DEXA). DEXA can non-invasively measure body composition, particularly total body fat and / or regional body fat mass. MRI can also be used to non-invasively measure body composition.

[0382] For all methods described herein, reference to the compounds of the present invention also includes compositions, such as pharmaceutical compositions described herein, that contain one or more of these compounds. These compositions may further include suitable excipients, such as pharma- ceutically acceptable excipients, including, for example, buffers known in the art. The present invention can be used alone or in combination with other conventional therapeutic methods.

[0383] A subject in need of the treatment provided by the present invention may have (i.e., may be diagnosed with or suffer from) at least one obesity-induced or obesity-related comorbidity, i.e., diseases and other adverse health conditions associated with, exacerbated by, or caused by, being overweight or obese. In other embodiments, a subject may have at least two obesity-induced or obesity-related comorbidities.

[0384] Obesity-induced or obesity-related comorbidities include, but are not limited to, diabetes mellitus, non-insulin-dependent type II diabetes mellitus, impaired glucose tolerance, impaired fasting glucose, dysglycaemia, elevated plasma insulin concentration, insulin resistance syndrome, hyperlipidemia, dyslipidemia, elevated free fatty acids, hypertension, hyperuricemia, gout, coronary artery disease, heart disease, myocardial infarction, angina pectoris, microvascular disease, sleep apnea, obstructive sleep apnea, Pickwickian syndrome, fatty liver; cerebral infarction, stroke, cerebral thrombosis, respiratory complications, cholelithiasis, gallbladder disease, kidney disease, gastroesophageal reflux, stress urinary incontinence, arteriosclerosis, heart disease, heart rhythm abnormalities, arrhythmias, transient ischemic attacks, orthopedic disorders, osteoarthritis, osteoarthritis, lumbodynia, menstrual disorders, hormonal imbalances, endocrine disorders, and infertility. In particular, comorbidities include: hypertension, hyperlipidemia, dyslipidemia, impaired glucose tolerance, cardiovascular disease, sleep apnea, diabetes mellitus, and other obesity-related conditions.

[0385] The present invention provides a method of treating obesity or inducing, causing or increasing weight loss (reducing body weight) in a subject in need thereof, as well as treating one or more of these obesity-induced or obesity-related comorbidities in a subject suffering from said comorbidities, said method comprising administering to the subject a therapeutically effective amount of at least one compound of the present invention to treat or ameliorate obesity or reduce body weight and treat or ameliorate one or more of the obesity-induced or obesity-related comorbidities.

[0386] The present invention provides a method of treating metabolic disorders or metabolic syndrome in a subject in need thereof, the syndrome being characterized by a group of metabolic risk factors including: 1) abdominal obesity (excess adipose tissue in and around the abdomen); 2) atherogenic dyslipidemia (high triglycerides; low HDL cholesterol and high LDL cholesterol, or low HDL:LDL ratio); 3) elevated blood pressure; 4) insulin resistance or impaired glucose tolerance; 5) prothrombotic state (e.g., high fibrinogen or high plasminogen activator inhibitor-1 in the blood); 6) proinflammatory state (e.g., elevated CRP in the blood); and 7) prediabetes or type 2 diabetes. The present invention can treat metabolic disease alone or in combination with treating obesity or inducing, causing or increasing weight loss.

[0387] The present invention also provides a method for treating, reducing or improving one or more cardiometabolic risk factors selected from the group consisting of, but not limited to, plasma triglyceride levels, LDL-cholesterol levels, C-reactive protein (CRP) levels and blood pressure (systolic and / or diastolic blood pressure) in a subject suffering from said risk factors, in addition to treating obesity or inducing, causing or increasing weight loss in a subject in need thereof, said method comprising administering to the subject a therapeutically effective amount of at least one compound of the present invention to treat or ameliorate obesity or reduce weight and treat or ameliorate one or more risk factors.

[0388] The compounds of the invention, or pharma- ceutically acceptable salts, prodrugs, metabolites, analogs or derivatives thereof, can also be administered in combination with a second active agent, which may be conjugated to a polymer.

[0389] Contemplated second active agents include those administered to treat type 2 diabetes, such as: sulfonylureas (e.g., chlorpropamide, glipizide, glyburide, glimepiride); meglitinides (e.g., repaglinide and nateglinide); biguanides (e.g., metformin); thiazolidinediones (rosiglitazone, troglitazone, and pioglitazone); glucagon-like 1 peptidomimetics (e.g., exenatide and liraglutide); sodium-glucose cotransporter inhibitors (e.g., dapagliflozin), dipeptidyl peptidase 4 inhibitors (e.g., gliptins), sodium-glucose ligated transporter inhibitors, renin inhibitors, agents, and alpha-glucosidase inhibitors (e.g., acarbose and meglitol); and / or those administered to treat cardiac disorders and conditions associated with overweight or obesity, such as hypertension, dyslipidemia, ischemic heart disease, cardiomyopathy, myocardial infarction, stroke, venous thromboembolic disease, and pulmonary hypertension, e.g., chlorthalidone; hydrochlorothiazide; indapamide, metolazone; loop diuretics (e.g., bumetanide, ethacrynic acid, furosemide, lasix, torsemide); potassium-sparing agents (e.g., amiloride hydrochloride, spironolactone, and triamterene); peripheral agents (e.g., reserpine); central alpha agonists (e.g., chlorthalidone, hydrochlorothiazide, indapamide, metolazone); alpha-agonists (e.g., clonidine hydrochloride, guanabenzacetate, guanfacine hydrochloride, and methyldopa); alpha-blockers (e.g., doxazosin mesylate, prazosin hydrochloride, and terazosin hydrochloride); beta-blockers (e.g., acebutolol, atenolol, betaxolol, bisoprolol fumarate, carteolol hydrochloride, metoprolol tartrate, metoprolol succinate, nadolol, penbutolol sulfate, pindolol, dolol, propranolol hydrochloride, and pyrilamine maleate); combinations of alpha-blockers and beta-blockers (e.g., carvedilol and labetalol hydrochloride); direct vasodilators (e.g., hydralazine hydrochloride and minoxidil); calcium antagonists (e.g., diltiazem hydrochloride and verapamil hydrochloride); dihydropyridines (e.g., amlodipine besylate, felodipine, isradipine, nicardipine, nifedipine, and nisoldipine);ACE inhibitors (benazepril hydrochloride, captopril, enalapril maleate, fosinopril sodium, lisinopril, moexipril, quinapril hydrochloride, ramipril, trandolapril); angiotensin II receptor blockers (e.g., losartan potassium, valsartan, and irbesartan); and combinations thereof; and statins, typically for the treatment of dyslipidemia, such as mevastatin, lovastatin, pravastatin, simvastatin, velostatin, dihydrocompactin, fluvastatin, atorvastatin, dalvastatin, carvastatin, crilvastatin, bevastatin, cefvastatin, rosuvastatin, pitavastatin, and glenvastatin;

[0390] Other second active agents that may be co-administered (e.g., sequentially or simultaneously) include: agents administered to treat ischemic heart disease, including statins, nitrates (e.g., isosorbide dinitrate and isosorbide dinitrate), beta-blockers, and calcium channel antagonists; agents administered to treat cardiomyopathies, including positive inotropes (e.g., digoxin), diuretics (e.g., furosemide), ACE inhibitors, calcium antagonists, antiarrhythmics (e.g., Sotolol, amiodarone, and disopyramide), and beta-blockers; agents administered to treat cardiomyopathies, including ACE inhibitors, angiotensin II receptor blockers, direct vasodilators, beta-blockers, antiarrhythmics, and thrombolytics (e.g., alteplase, Retaplase, tenecteplase, anistrephrase, thrombopenia ... agents administered to treat myocardial infarction, including antiplatelet agents (e.g., aspirin, clopidogrel, dipyridamole, and ticlopidine), anticoagulants (e.g., heparin), and thrombolytic agents; agents administered to treat venous thromboembolic disease, including antiplatelet agents, anticoagulants, and thrombolytic agents; agents administered to treat pulmonary hypertension, including positive inotropes, anticoagulants, diuretics, potassium (e.g., K-dur), vasodilators (e.g., nifedipine and diltiazem), bosentan, epoprostenol, and sildenafil; agents administered to treat asthma, including bronchodilators, anti-inflammatory agents, leukotriene blockers, and anti-IgE agents. Certain asthma medications include zafirlukast, flunisolide, triamcinolone, beclomethasone, terbutaline, fluticasone, formoterol, beclomethasone, salmeterol, theophylline, and Xopenex; medications administered to treat sleep apnea include modafinil and amphetamines; medications administered to treat non-alcoholic fatty liver disease include antioxidants (e.g., vitamins E and C), insulin sensitizers (metformin, pioglitazone, rosiglitazone, and betaine), hepatoprotectants, and lipid-lowering medications;Medications administered to treat osteoarthritis of weight-bearing joints include acetaminophen, nonsteroidal anti-inflammatory drugs (e.g., ibuprofen, etodolac, oxaprozin, naproxen, diclofenac, and nabumetone), COX-2 inhibitors (e.g., celecoxib), steroids, supplements (e.g., glucosamine and chondroitin sulfate), and artificial joint fluid; medications administered to treat Prader-Willi syndrome include human growth hormone (HGH), somatropin, and antiobesity drugs (e.g., orlistat, sibutramine, and rituximab). drugs administered to treat polycystic ovary syndrome include insulin sensitizers, synthetic estrogen and progesterone combinations, spironolactone, eflornithine, and clomiphene; drugs administered to treat erectile dysfunction include phosphodiesterase inhibitors (e.g., tadalafil, sildenafil citrate, squalene, methamphetamine, ionamine, phentermine, bupropion, diethylpropion, phendimetrazine, benzphetermine, and topamax); drugs administered to treat polycystic ovary syndrome include insulin sensitizers, synthetic estrogen and progesterone combinations, spironolactone, eflornithine, and clomiphene; drugs administered to treat erectile dysfunction include phosphodiesterase inhibitors (e.g., tadalafil, sildenafil citrate, squalene, methamphetamine ... Drugs administered to treat infertility include clomiphene, clomiphene citrate, bromocriptine, gonadotropin releasing hormone (GnRH), GnRH agonists, GnRH antagonists, tamoxifen / Nolvadex, gonadotropins, human chorionic gonadotropin (HCG), human menopausal gonadotropin (HmG), progesterone, recombinant follicle-stimulating hormone (RFH), ovarian hypertension, ... follitropin, heparin, follitropin alpha, and follitropin beta;medications administered to treat labor complications include bupivacaine hydrochloride, dinoprostone PGE2, meperidine HC1, Ferro-folic-500 / iberet-folic-500, meperidine, methylergonovine maleate, ropivacaine HC1, nalbuphine HC1, oxymorphone HC1, oxytocin, dinoprostone, ritodrine, scopolamine hydrobromide, sufentanil citrate, and labor-inducing drugs;Medications administered to treat depression include serotonin reuptake inhibitors (e.g., fluoxetine, escitalopram, citalopram, paroxetine, sertraline, and venlafaxine); tricyclic antidepressants (e.g., amitriptyline, amoxapine, clomipramine, desipramine, dosulepin hydrochloride, doxepin, imipramine, iprindole, lofepramine, nortriptyline, opipramol, protriptyline, and trimipramine); monoamine oxidase inhibitors (e.g., isocarboxazid, moclobemide, phenelzine, tranylcypromine, selegiline, rasagiline, nialamide, iproniazid, iproclozide, toloxatone, linezolid, dienolide kavapyrone, desmethoxyyangonine, desmethoxyyangonin, and dextroamphetamine); stimulants (e.g., amphetamine, methamphetamine, methylphenidate, and arecoline); antipsychotics (e.g., butyrophenones, phenothiazines, thioxanthenes, clozapine, olanzapine, risperidone, quetiapine, ziprasidone, amisulpride, paliperidone, symbiakis, tetrabenazine, and cannabidiol); and mood stabilizers (e.g., lithium carbonate, valproic acid, divalproex sodium, sodium valproate, lamotrigine, Drugs administered to treat anxiety include serotonin reuptake inhibitors, mood stabilizers, benzodiazepines (e.g., alprazolam, clonazepam, diazepam, and lorazepam), tricyclic antidepressants, monoamine oxidase inhibitors, and beta-blockers; and other anti-obesity drugs, including: serotonin and noradrenaline reuptake inhibitors; noradrenaline reuptake inhibitors; selective serotonin reuptake inhibitors; and intestinal lipase inhibitors. Specific anti-obesity drugs include orlistat, sibutramine, methamphetamine, ionamine, phentermine, bupropion, diethylpropion, phendimetrazine, benzphetermine, and topamax.

[0391] The present invention also provides a method for reducing adipocytes in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of at least one compound of the present invention to reduce adipocytes or adipose tissue. The present invention also provides a method for preventing an increase in adipocytes in a subject at risk thereof, comprising administering to the subject a therapeutically effective amount of at least one compound of the present invention to prevent an increase in adipocytes. Reducing adipocytes means reducing the number or reducing the size (fat mass) of adipocytes. Preventing an increase in adipocytes means reducing or maintaining the number of adipocytes or reducing or maintaining the size of adipocytes. In some embodiments, administering a compound of the present invention shrinks adipocytes in a subject in need thereof. The adipocytes can be white adipose tissue or brown adipose tissue.

[0392] The present invention also provides a method for reducing food intake in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of at least one compound of the present invention to reduce food intake.

[0393] A reduction in food intake means a reduction in daily food intake. The reduction in daily food intake can be about 5% to about 50% (e.g., about 5%, about 10%, about 20%, about 30%, about 40%, or about 50%). Based on a daily diet of 2000 kcal, the reduction is about 100 kcal to about 1000 kcal per day (e.g., about 100 kcal, about 200 kcal, about 400 kcal, about 600 kcal, about 800 kcal, or about 1000 kcal).

[0394] The present invention also provides a method of reducing hunger in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of at least one compound of the present invention to reduce hunger. The subject may also have a reduced food intake.

[0395] Hunger can be assessed in the fasted state using a 10-point visual analog scale (VAS), which is commonly used in appetite studies. See Flint et al., Int. J. Obes. Relat. Metab. Disord. 24(1):38-48, 2000. Specifically, subjects are asked to rate their overall hunger over the previous two days on a scale of 1 to 10, with 10 being extremely hungry and 1 being not hungry at all.

[0396] The methods of the present invention can also reduce waist circumference in a subject in need thereof. Waist circumference is assessed using a tape measure placed abdominally 1 cm above the iliac crest. A subject of the present invention may have a reduction in waist circumference of about 1 inch (2.54 cm) to about 20 inches (50.8 cm) (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 inches (2.54 cm, 5.08 cm, 7.62 cm, 10.16 cm, 12.7 cm, 15.24 cm, 17.78 cm, 20.32 cm, 22.86 cm, 25.4 cm, 27.94 cm, 30.48 cm, 33.02 cm, 35.56 cm, 38.1 cm, 40.64 cm, 43.18 cm, 45.72 cm, 48.26 cm, or 50.8 cm).

[0397] In the method of the present invention, the administration of the compound results in a reduction in body fat and a substantial maintenance of muscle mass in the patient. In some embodiments, the administration of the compound promotes fat oxidation in the patient, compared to the patient who is only on a restricted food intake regimen. For example, provided herein is a method for reducing body fat in a patient in need thereof. Such a patient may maintain substantially more muscle mass, compared to the reduction in body fat in a patient who is only on an energy restricted diet regimen.

[0398] The present invention also provides a method for improving surgical outcomes in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of at least one compound of the present invention prior to surgery to improve surgical outcomes. In some embodiments, the administration reduces liver and / or abdominal fat in the patient and improves surgical outcomes. In some embodiments, the surgery is not an emergency surgery. Such surgery may include bariatric surgery, cardiovascular surgery, abdominal surgery, or orthopedic surgery.

[0399] A "patient" or "subject" as used herein can refer to a human or non-human subject. In some embodiments, the subject is a vertebrate. In some embodiments, the vertebrate is a mammal. Mammals include, but are not limited to, farm animals, sport animals, pets, primates (including humans), horses, dogs, cats, mice and rats. In some embodiments, the mammal is a human.

[0400] As used herein, a "subject in need thereof" is a subject who is overweight or obese (with or without one or more comorbidities), or who is at high risk relative to the general population for becoming overweight or developing obesity. In some embodiments, a subject in need thereof is one who is overweight or obese (with or without one or more comorbidities), or who is at high risk relative to the general population for becoming overweight or developing obesity. 2 In some embodiments, the subject in need thereof is a patient who is overweight or obese, or at high risk of becoming overweight or developing obesity relative to the general population, and who does not suffer from or has not been diagnosed with a disorder selected from the group consisting of cancer, hyperproliferative disorders, retinal neovascularization due to macular degeneration, psoriasis and pyogenic granuloma, rheumatoid, immune and osteoarthritis.

[0401] The term "prophylactic or therapeutic" treatment is art-recognized and includes administration of one or more of the subject compositions to a host. If the treatment is administered prior to clinical signs of an undesirable condition (e.g., a disease or other undesirable condition in a host animal), the treatment is prophylactic (i.e., protects the host from developing the undesirable condition), whereas if the treatment is administered after signs of an undesirable condition, the treatment is therapeutic (i.e., intended to reduce, ameliorate, or stabilize an existing undesirable condition or its side effects).

[0402] As used herein, "treatment" is an approach for obtaining beneficial or desired clinical results. For purposes of this invention, beneficial or desired clinical results include, but are not limited to, one or more of: improving one or more symptoms associated with a disease; reducing its severity; or alleviating it. With respect to obesity, beneficial or desired clinical outcomes include any one or more of the following: reducing or maintaining body weight; controlling (including reducing) food intake or caloric intake; increasing metabolic rate or inhibiting a decrease in metabolic rate; and improving, reducing the severity and / or alleviating any of the disorders associated with obesity, such as, for example, diabetes, non-insulin dependent diabetes mellitus, hyperglycemia, low glucose tolerance, insulin resistance, lipid disorders, dyslipidemia, hyperlipidemia, hypertriglyceridemia, hypercholesterolemia, abdominal obesity, eating disorders, metabolic syndrome, hypertension, osteoarthritis, myocardial infarction, fatty liver disease, steatohepatitis, non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver disease (NAFLD), stroke and other related diseases; improving the quality of life and / or extending the lifespan of patients with obesity.

[0403] As used herein, "delaying" the onset of obesity means to postpone, prevent, slow down, delay, stabilize, and / or postpone the onset of the disease. This delay can be of various lengths of time, depending on the history of the disease and / or medical history of the subject being treated. As will be apparent to one skilled in the art, a sufficient or significant delay can actually encompass prevention, in that the individual does not develop the disease. For example, one result of delaying onset can be to reduce the body weight of a subject at risk of obesity, compared to the body weight of the subject immediately before administration of the composition described herein. Another result of delaying onset can be to prevent the regain of weight previously lost as a result of diet, exercise, or drug therapy. Another result of delaying onset can be to prevent the onset of obesity in a subject at risk of obesity, if the treatment is administered before the onset of obesity. Another result of delaying onset can be to reduce the occurrence and / or severity of obesity-related disorders in a subject at risk of obesity, if the treatment is administered before the onset of obesity.

[0404] An individual "at risk" of obesity may or may not have detectable disease, and may or may not exhibit detectable disease prior to the implementation of the treatment methods described herein. "At risk" refers to an individual having one or more so-called risk factors, which are measurable parameters that correlate with the development of obesity. Individuals with one or more of these risk factors have a higher probability of being obese than individuals without these risk factors. These risk factors include, but are not limited to, age, diet, physical inactivity, metabolic syndrome, family history of obesity, ethnicity, genetic syndrome, previous medical history (e.g., eating disorders, metabolic syndrome, and obesity), and the presence of precursor diseases (e.g., overweight). For example, a person with a body weight of 25.0 to 30.0 kg / m 2 Otherwise healthy individuals with a BMI less than 25.0 kg / m 2 ~27.0kg / m 2 Individuals with at least one comorbidity who have a BMI below 100% are at risk for obesity.

[0405] "Development" of obesity refers to the onset and / or progression of the disease in an individual, which may be different embodiments of the present invention. Onset of obesity can be detected using standard clinical techniques described herein. Onset may also refer to the progression of the disease, which is not detectable in the early stages. For the purposes of the present invention, progression refers to the biological process of the disease state, where progression is measured by height and weight assessment to estimate BMI, measuring waist circumference, assessing comorbidities, and the onset and / or worsening of obesity complications, such as arteriosclerosis, type II diabetes, polycystic ovarian disease, cardiovascular disease, osteoarthritis, dermatological disorders, hypertension, insulin resistance, hypercholesterolemia, hypertriglyceridemia, and cholelithiasis. These various diagnostic tests are known in the art. "Onset" includes onset, recurrence, and onset. As used herein, "onset" or "onset" of obesity includes initial onset and / or recurrence.

[0406] As used herein, "body weight control" or "body weight improvement" refers to reducing or maintaining body weight in an individual (compared to pre-treatment levels). In some embodiments, body weight is maintained within a generally normal range. Body weight can be reduced by reducing caloric intake and / or reducing body fat stores. In some embodiments, body weight is reduced in an individual by at least about any of 3%, 4%, 5%, 10%, 20%, 30%, 40%, or 50% compared to pre-treatment levels.

[0407] As used herein, "controlling food intake" refers to reducing or maintaining food intake in an individual (compared to pre-treatment levels). In some embodiments, food intake is maintained in a generally normal range. In some embodiments, food intake is reduced by about any of 3%, 4%, 5%, 10%, 20%, 30%, 40%, or 50% in an individual compared to pre-treatment levels.

[0408] A "therapeutically effective amount" of a compound, with respect to use in therapy, refers to the amount of compound in a formulation that, when administered as part of a desired dosing regimen (to a mammal, preferably a human), alleviates the symptoms, ameliorates the condition, or delays or prevents the onset of a disease state, according to clinically acceptable criteria for the disorder or condition being treated or for cosmetic purposes, such as a reasonable benefit / risk ratio applicable to any medical treatment. A "therapeutically effective amount" is synonymous with an "effective dose."

[0409] As used herein, an "effective dosage" or "effective amount" of a drug, compound, or pharmaceutical composition is an amount sufficient to produce a beneficial or desired result. In prophylactic use, beneficial or desired results include results such as eliminating or reducing the risk of, reducing the severity, or delaying the onset of a disease, including biochemical, histological, and / or behavioral signs of the disease, complications of the disease, and intermediate pathological phenotypes manifested during the development of the disease. In therapeutic use, beneficial or desired results include clinical results such as, for example, reducing the intensity, duration, or frequency of attacks of the disease; and reducing one or more signs (biochemical, histological, and / or behavioral) resulting from the disease, including complications of the disease and intermediate pathological phenotypes manifested during the development of the disease; increasing the quality of life of a patient suffering from the disease; reducing the dose of other drugs required to treat the disease; enhancing the effect of another drug therapy; and / or delaying the progression of the disease in a patient. An effective dosage can be administered in one or more administrations. For the purposes of the present invention, an effective dosage of a drug, compound, or pharmaceutical composition is an amount sufficient to achieve prophylactic or therapeutic treatment, either directly or indirectly. As understood in a clinical context, an effective dosage of a drug, compound, or pharmaceutical composition may or may not be achieved in conjunction with another drug, compound, or pharmaceutical composition. Thus, when a desired result can or is achieved in combination with one or more other drugs, an "effective dosage" may be considered in the context of administering one or more therapeutic agents, and may also be considered to administer a single drug in an effective amount. For example, an effective amount of a compound of the present invention for treating obesity is an amount sufficient to treat or ameliorate one or more symptoms associated with obesity.An "effective amount" is an amount sufficient to result in one or more of the following (which may correspond to various embodiments of the present invention): reducing, reducing or controlling body weight; reducing, reducing or controlling food intake; increasing metabolic rate; reducing one or more symptoms resulting from diseases associated with obesity; increasing the quality of life of a patient suffering from obesity; and / or extending lifespan.

[0410] When providing one or more compounds described herein to a subject, the dosage of the compound administered will vary depending on factors such as the subject's age, weight, height, sex, general medical condition, previous medical history, disease progression, route of administration, formulation, and the like.

[0411] The dosage of the compound of the present invention can be empirically determined in an individual who has received one or more doses. The individual is administered increasing doses of the compound of the present invention. To evaluate the effectiveness of the compound of the present invention, markers of disease state can be monitored. It will be clear to those skilled in the art that dosage will vary depending on the individual, stage of disease (e.g., stage of obesity), and previous and concurrent treatments.

[0412] The toxicity and therapeutic efficacy of the compounds of the present invention can be determined by standard pharmaceutical procedures in experimental animals. The toxic dose can be determined as the maximum tolerated dose (MTD) or LD50 (the dose lethal to 50% of the population). The effective dose can be determined as the ED50 (the dose therapeutically effective in 50% of the population) or the dose required to produce some mean change in animals (e.g., the dose required to produce a mean reduction in systolic blood pressure of 10 mmHg in a group of subjects).

[0413] Ideally, the effective dose and the toxic dose are determined in the same species. However, when they are determined in different species, allometric scaling can be used to convert to the effective or toxic dose for the other species. The dose ratio of toxic to therapeutic effects is the therapeutic index, which can be expressed as the LD50 / ED50 ratio. When comparing mice to rats, a commonly accepted scaling factor is 2, with the dose in rats estimated to be half that in mice. Thus, if the toxic dose in rats is 100 mg / kg and the effective dose in mice is 1 mg / kg, the therapeutic index in rats can be calculated as the effective dose in rats is 1 mg / kg / 2, or 0.5 mg / kg, resulting in a therapeutic index of 200. The FDA defines a drug as having a narrow therapeutic window if (a) the difference between the median lethal dose and the median effective dose is less than two-fold; or (b) the difference between the minimum toxic concentration and the minimum effective concentration in the blood is less than two-fold.

[0414] Compounds of the present invention that exhibit high therapeutic indices are preferred. Compounds of the present invention that exhibit toxic side effects can also be used, but care should be taken in designing delivery systems that deliver such compounds of the present invention to the site of affected tissues in order to minimize the possibility of damaging non-infected cells, thereby reducing side effects.

[0415] Data obtained from animal studies can be used to formulate a range of dosages for use in humans. The dosage of such compounds of the invention is preferably a range of blood concentrations that includes the effective dose with little or no toxicity. The dosage can vary within this range depending on the dosage form used and the route of administration utilized. For compounds of the invention with MW less than 1000, the therapeutically effective dose can be initially estimated from cell culture assays, while animal models allow better estimation of doses for conjugates that require linker cleavage to release the active moiety. Such information can be used to more accurately determine useful doses in humans. It is known in the art that conjugation with a polymer dilutes the active moiety activity (the polymer is a diluent). This is illustrated in the mouse dosing model of anticancer drugs shown in the table below.

[0416] [Table 2]

[0417] It is therefore well understood that conjugation with polymers increases the clinical dose if not improves the therapeutic index, as exemplified in the human dosing models of anticancer drugs shown in the table below.

[0418] [Table 3]

[0419] The polymer-conjugated and modified compounds of the present invention surprisingly provide greater efficacy and lower toxicity compared to the unconjugated and / or unmodified parent drug / active moiety.

[0420] For example, the fumagillol complex and modified fumagillol compound of the present invention are surprisingly superior to fumagillol small molecule, and at equimolar doses, they cause increased weight loss in DIO mice.The compounds of the present invention can be used at lower molar doses and less frequent dosing to cause equivalent weight loss.Lower molar doses and reduced dosing frequency reduce systemic drug exposure and systemic drug toxicity.Furthermore, the fumagillol complex and modified fumagillol compound of the present invention provide effects similar to those of fumagillol small molecule, i.e., favorable fat loss and reduced food intake in DIO mice.

[0421] Conventional polymer conjugates dilute activity, increase doses 5-20 fold, and produce only modest changes in therapeutic index (less than 2-fold). In contrast, the polymer conjugated compounds of the present invention surprisingly and unexpectedly increase (greatly improve) the therapeutic index and show increased activity at lower doses.

[0422] In the methods of the present invention, the polymer conjugated compounds of the present invention exhibit less frequent dosing (e.g., q4d: dosing every 4 days, q7d: dosing every 7 days, q8d: dosing every 8 days), reduced doses to at least 84 molar % of fumagillol equivalents, reduced AUC in non-target compartments while the therapeutic index is increased (more than 10-fold).

[0423] In another embodiment, effective dosage, for example, daily dosage of the compound of the present invention is provided herein.For example, a method is provided herein, which comprises administering a dose of the compound of the present invention that is effective for weight loss.For example, in the method described herein, the administration of the compound of the present invention contemplated can comprise administering a dose of about 200mg / day, about 80mg / day, about 40mg / day, about 20mg / day, about 10mg / day, about 5mg / day, about 3mg / day, about 2mg / day, about 1mg / day, about 0.5mg / day, about 0.2mg / day, about 0.05mg / day, about 0.01mg / day, or about 0.001mg / day, regardless of body weight.

[0424] An effective amount of the drug for weight loss in a patient may be administered based on body weight or body surface area and may be about 0.0001 mg / kg to about 5 mg / kg body weight / day. For example, contemplated dosages may be about 0.001 to 5 mg / kg body weight / day, about 0.001 mg / kg to 1 mg / kg body weight / day, about 0.001 mg / kg to 0.1 mg / kg body weight / day, about 0.001 to about 0.010 mg / kg body weight / day, or about 0.007 mg / kg body weight / day.

[0425] The compounds of the invention can be administered in an amount sufficient to reduce the patient's body weight by about 0.5 kg / week to about 1 kg / week (or about 0.5% of body weight per week to about 1% of body weight per week). In some embodiments, the reduction in weekly body weight occurs for the duration of treatment.

[0426] Administration of the compounds of the invention according to the methods of the invention may be continuous or intermittent, depending, for example, on the physiological condition of the patient receiving the administration, whether the purpose of the administration is therapeutic or prophylactic, and other factors known to the skilled practitioner. Administration of the compounds of the invention may be essentially continuous over a preselected period of time, or may be in a series of spaced doses.

[0427] For repeated administration over several days or longer, depending on the condition, treatment is continued until the desired suppression of symptoms or sufficient therapeutic levels are achieved. For example, dosing 1-5 times per week is contemplated. In certain embodiments, the compounds of the invention are administered about every 4 days. Other dosing regimes include 1-5 times per week, every 3-4 days, or less frequent regimes. In some embodiments, the compounds of the invention are administered about once per week, once every 2 weeks, or about 1-4 times per month, depending on the duration of response to drug administration. Intermittent dosing regimes with staggered dosages spaced 2 days up to 7 days or 14 days apart may be used. In some embodiments, treatment may begin with daily dosing, then change to weekly to monthly dosing. The progress of this treatment can be easily monitored by conventional techniques and assays, or by measuring MetAP2 as described in U.S. Pat. No. 6,548,477.

[0428] The frequency of administration can be determined and adjusted over the course of treatment.For example, the frequency of administration can be determined or adjusted based on the type and severity of the disease being treated, whether the purpose of administration of the drug is preventive or therapeutic, previous treatment history, the clinical history of the patient, and response to the drug, and the instructions of the attending physician.Typically, the clinician administers the compound of the present invention until a dosage that achieves the desired result is reached.

[0429] Treatment can continue for as long as desired or can be limited to a short period of time. Suitable treatment periods can be, for example, at least about 1 week, at least about 4 weeks, at least about 1 month, at least about 6 months, at least about 1 year, at least about 2 years, or indefinitely. Treatment periods can end when desired results, such as targeted weight loss, are achieved, for example, about 5% of body weight, about 10% of body weight, about 20% of body weight, about 30% of body weight or more are achieved. Treatment regimes can include: a correction phase, in which the compound of the present invention is administered at a dose or dosage frequency sufficient to cause a reduction in excess fat accumulation; followed by a maintenance phase, in which a lower compound dose or dosage frequency sufficient to prevent the recurrence of excess fat accumulation is administered.

[0430] The compound or its pharma- ceutically acceptable salt, ester or prodrug (or pharmaceutical composition thereof) can be administered by any means known in the art. For example, the compound or composition of the present invention can be administered orally, nasally, transdermally, topically, pulmonary, inhalation, buccal, sublingually, intraperitoneally, subcutaneously, intramuscularly, intravenously, rectally, intrathoracically, intrathecally and parenterally. Administration can be systemic, such as intravenous, or local. In some embodiments, the route of administration can be intravenous, intramuscular, subcutaneous, intradermal, intraperitoneal, intrathecal, intrathoracic, intrauterine, rectal, vaginal, topical, and the like. In some embodiments, the compound is administered subcutaneously.

[0431] In one aspect, the compounds of the present invention, or pharma- ceutically acceptable salts, solvates, diastereomers, and polymorphs thereof, can be administered in suitable dosage forms or formulations prepared according to conventional procedures by combining a therapeutically effective amount (e.g., an effective level sufficient to achieve the desired therapeutic effect) of a compound of the present invention, or a pharma- ceutically acceptable salt, solvate, diastereomer, and polymorph thereof (as an active ingredient) with a standard pharmaceutical carrier or diluent (i.e., by making a pharmaceutical composition of the present invention). These procedures may involve mixing, granulating, and compressing or dissolving the ingredients, as appropriate to achieve the desired formulation.

[0432] Parenteral dosage forms can be prepared by any means known in the art. For example, a sterile injectable aqueous or oleaginous suspension may be formulated according to known techniques using suitable dispersing or wetting agents and suspending agents.

[0433] Oral dosage forms, such as capsules, tablets, pills, powders, and granules, can be prepared by any suitable method known in the art.For example, the compound of the present invention can be mixed with an enteric material and compressed into a tablet.Alternatively, the formulation of the present invention can be incorporated into a chewable tablet, a crushable tablet, a tablet that dissolves quickly in the mouth, or a mouthwash.

[0434] For pulmonary (e.g., intrabronchial) administration, the compounds of the present invention can be formulated with conventional excipients to prepare inhalable compositions in the form of fine powder or nebulizable liquid. For ocular administration, the compounds of the present invention can be formulated with conventional excipients to form, for example, eye drops or eye implants. Excipients useful in eye drops include thickeners or gelling agents to improve retention in the eye and thereby minimize loss due to tearing.

[0435] Liquid dosage forms for oral or other administration include, but are not limited to, pharma- ceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups, and elixirs. In addition to the active agent, liquid dosage forms may contain inert diluents commonly used in the art, such as water or other solvents, solubilizers and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (more specifically, cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil, and sesame oil), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycol, and fatty acid esters of sorbitan, and mixtures thereof. In addition to inert diluents, compositions for ocular delivery, oral delivery, or other systemic delivery may contain auxiliary agents, such as wetting agents, emulsifiers, and suspending agents.

[0436] Commercially available nebulizers for liquid formulations, including jet nebulizers and ultrasonic nebulizers, are useful for administration.Liquid formulations can be directly nebulized, and lyophilized powders can be nebulized after reconstitution.Alternatively, the compounds of the present invention can be aerosolized using fluorocarbon formulations and metered dose inhalers, or inhaled as lyophilized and milled powders.

[0437] The dosage form for topical or transdermal administration of the pharmaceutical composition of the present invention may include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants, or patches.The active agent is mixed under sterile conditions with a pharma- ceutically acceptable carrier and any preservatives or buffers that may be required.For example, administration via the dermal route is achieved by aqueous drops, sprays, emulsions, or creams.

[0438] Transdermal patches may have the additional advantage of providing controlled delivery of active ingredients to the body. Such dosage forms can be made by dissolving or dispensing the compound in a suitable medium. Absorption enhancers may be used to increase the flux of the compound across the skin. The rate can also be controlled by providing a rate-controlling membrane or by dispersing the compound in a polymer matrix or gel.

[0439] Compositions for rectal or vaginal administration may be suppositories, which may be prepared by mixing the compounds of the present invention with suitable non-irritating excipients or carriers, such as cocoa butter, polyethylene glycol, or suppository waxes, which are solid at ambient temperature but liquid at body temperature and thus melt in the rectum or vaginal cavity to release the active agent. Alternatively, administration may be by inserting an endoscope into the subject's rectum followed by release of the intended formulation from the lumen of the endoscope.

[0440] Those skilled in the art may refer to general references for detailed descriptions of known techniques or equivalent techniques described herein. These references include Ausubel et al., Current Protocols in Molecular Biology; John Wiley and Sons, Inc. (2005); Sambrook et al., Molecular Cloning, A Laboratory Manual (3d ed.), Cold Spring Harbor Press, Cold Spring Harbor, New York (2000); Coligan et al., Current Protocols in Immunology, John Wiley & Sons, NY; Enna et al., Current Protocols in Pharmacology, John Wiley & Sons, NY; Fingl et al., The Pharmacological Basis of Therapeutics (1975), Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, PA, 18th edition (1990); Of course, these references may also be used in the preparation and use of the embodiments of the present invention. EXAMPLES

[0441] Examples are provided below to further illustrate various features of the present invention. The examples also illustrate useful methodologies for practicing the present invention. These examples do not limit the invention as claimed.

[0442] Basic steps Tangential flow filtration (TFF) was used to purify the polymer products of the invention. TFF was performed using a Pall Minimate™ Capsule and a Minimate™ TFF system according to the manufacturer's instructions. A Minimate TFF Capsule equipped with a 5 kDa Omega membrane (5K) or a Minimate TFF Capsule equipped with a 10 kDa Omega membrane (10K) cartridge was used for purification. In all cases, the permeate was discarded and the retentate was lyophilized to obtain the polymer product. The structure of the product was 1 The polymers were confirmed by H NMR and characterized by mass spectrometry (MS). Polymer weights reported in the examples were not corrected for water content.

[0443] Carbamoyl fumagillol and chloroacetylcarbamoyl fumagillol can be prepared according to the method disclosed in U.S. Patent No. 5,166,172 (Kishimoto et al., which is incorporated herein by reference). p-Nitrophenyl fumagyl-6-yl carbonate can be prepared according to published procedures. (See Han, C. et al., Biorg. Med. Chem. Lett. 2000, 10, 39-43.) MA-GFLG-ONp can be prepared according to the method disclosed in U.S. Patent No. 5,258,453 (Kopecek et al., which is incorporated herein by reference).

[0444] Synthesis of poly(HPMA-co-MA-GFLG-ONp)

[0445] [ka]

[0446] A mixture of hydroxypropyl methacrylamide (HPMA, 22.16 g, 155 mmol), N-methyacryl-gly-phe-leu-gly p-nitrophenyl ester (MA-GFLG-ONp, 10.00 g, 17.19 mmol), AIBN (1.484 g, 9.037 mmol) and acetone (225 g) was degassed (four cycles of freeze, pump and thaw). The resulting reaction mixture was stirred at 50° C. for 48 h and then cooled to room temperature. The desired product was purified by trituration with acetone and then dried under vacuum to give 17.6 g of poly(HPMA-co-MA-GFLG-ONp) as a white solid. The structure was 1 H NMR showed that the product was essentially free of impurities (e.g., p-nitrophenol). UV absorbance showed that the copolymer contained 0.47 mmol of p-nitrophenyl ester per gram of polymer. This copolymer is used in most of the subsequent examples. A wide variety of copolymers based on different monomers and / or monomer ratios may be made following this procedure by adjusting the stoichiometry and / or using different monomers.

[0447] Synthesis of poly(HPMA-co-MA-GFLG-OH)

[0448] Poly(HPMA-co-MA-GFLG-ONp) (700 mg) was added portionwise to a solution of 0.1 M NaOH (11.3 mL) at 0° C. The yellow reaction mixture was stirred at 0° C. for 0.5 h and then at room temperature for 4 h. Half of the solution was acidified to pH=6 with 0.1 M HCl. The aqueous phase was extracted with ethyl acetate to remove excess p-nitrophenol. The aqueous phase was lyophilized to give poly(HPMA-co-MA-GFLG-OH) as a colorless solid (360 mg).

[0449] Synthesis of poly(HPMA-co-MA-GG-ONp)

[0450] [ka]

[0451] A mixture of hydroxypropyl methacrylamide (HPMA, 82.5 g), N-methylacryl-gly-gly p-nitrophenyl ester (MA-GG-ONp, 16.8 g), AIBN (5.7 g), and acetone (875 g) was bubbled with argon for 90 min. The resulting reaction mixture was stirred at 50 °C for 48 h and then cooled to room temperature. The desired product was purified by trituration with acetone and then dried in vacuum to give 69.3 g of poly(HPMA-co-MA-GG-ONp) as a white solid. The structure was 1 H NMR confirmed that the product was substantially free of impurities (e.g., p-nitrophenol). The amount of p-nitrophenyl ester per gram of polymer may be determined by UV absorbance. A wide variety of copolymers based on different monomers and / or monomer ratios may be made following this procedure by adjusting the stoichiometry and / or using different monomers.

[0452] Poly(HPMA-co-MA-GFLG-NHCH 2 CH 2 Synthesis of N(Me)BOC) and General Procedure A

[0453] [ka]

[0454] DMF (6 mL) and H 2A solution of poly(HPMA-co-MA-GFLG-ONp) (1.0 g, 0.534 mmol) in O (10 mL) was added dropwise to a solution of tert-butyl N-(2-aminoethyl)-N-methylcarbamate (0.20 g, 1.15 mmol) in water (20 mL) at 0° C. in 15 min. The reaction mixture was stirred at 0° C. for 15 min, then warmed to room temperature and stirred for 12 h. The solvent was evaporated under reduced pressure. The resulting residue was dissolved in water (50 mL) and the pH was adjusted to about 8.0 with 0.1 M NaOH. The solution was filtered through a VacuCap filter and then purified using TFF (10K). The polymer-containing solution was washed (as part of the TFF process) with 25 mM NaCl solution (800 mL) to remove p-nitrophenol, the pH of the solution was adjusted to approximately 4 with 0.1 M HCl, and then washed (as part of the TFF process) with water (400 mL). The polymer solution was lyophilized to give the compound poly(HPMA-co-MA-GFLG-NHCH 2 CH 2 N(Me)BOC) was isolated as a pale yellow solid (720 mg, 71%).

[0455] Fmoc-Phe-Gly-NH-(CH 2 ) 6 Synthesis of NH-Boc: A solution of Fmoc-Phe-Gly-OH (0.66 g) in anhydrous THF (20 mL) was diluted with N 2 N,N'-dicyclohexylcarbodiimide (0.307 g) and 1-hydroxybenzotriazole hydrate (0.201 g) were added at 0°C. After stirring for 15 min, N-Boc-1,6-diaminohexane (0.322 g) was added. The reaction mixture was allowed to warm to room temperature and stirred overnight. The solids were filtered off and washed with EtOAc. The filtrate and washings were then concentrated under reduced pressure. The resulting residue was purified by flash column chromatography (CH 2 Cl 2 Purification with 0–10% MeOH in 10 mL of 1000 mL of Fmoc-Phe-Gly-NH-(CH 2 ) 6 NH-Boc was obtained as a white solid (0.9 g).

[0456] Fmoc-Phe-Gly-NH-(CH2 ) 6 NH 2 Synthesis of -TFA: Fmoc-Phe-Gly-NH-(CH 2 ) 6 NH-Boc (0.7 g) was dissolved in CH 2 Cl 2 (4mL) in N 2 The mixture was dissolved at 0° C. and then trifluoroacetic acid (TFA) (4 mL) was added. The reaction mixture was allowed to warm to room temperature and then N 2 The mixture was stirred at RT for 2 h. The solvent was removed under reduced pressure, and the residue was dried under high vacuum to give 0.71 g of Fmoc-Phe-Gly-NH(CH 2 ) 6 -NH 2 TFA was obtained. This crude material was used for preparation without further purification.

[0457] Fmoc-Phe-Gly-NH(CH 2 ) 6 Synthesis of NH-CO-fumagillol:

[0458] [ka]

[0459] N 2 Bottom, anhydride CH 2 Cl 2 Compound Fmoc-Phe-Gly-NH(CH 2 ) 6 To a solution of -NH2TFA (0.71 g) at 0°C, nitrophenyl fumagyl-6-yl carbonate (0.536 g) was added. Diisopropylethylamine (diPEA) (0.74 mL) was then added. The reaction mixture was allowed to warm to room temperature and then stirred at the same temperature overnight. The solvent was removed under reduced pressure and the resulting residue was dissolved in EtOAc (70 mL). The EtOAc was washed with water and brine. The ethyl acetate solution was then diluted with MgSO 4 The residue was purified by flash column chromatography (CH 2 Cl 2Purification with 0–10% MeOH in 10 mL of 1000 mL of Fmoc-Phe-Gly-NH-(CH 2 ) 6 NH-CO-fumagillol was obtained as an off-white solid (0.81 g).

[0460] H-Phe-Gly-NH(CH 2 ) 6 Synthesis of NH-CO-fumagillol

[0461] [ka]

[0462] N 2 Bottom, anhydride CH 2 Cl 2 (20mL) in compound Fmoc-Phe-Gly-NH-(CH 2 ) 6 To a solution of NH-CO-fumagillol (0.80 g) at 0° C. was added DBU (0.15 g). The reaction mixture was allowed to warm to room temperature. The solvent was removed under reduced pressure and the resulting residue was purified by flash column chromatography (CH 2 C1 2 H-Phe-Gly-NH-(CH 2 ) 6 H-CO-fumagillol was obtained as a pale yellow gum (0.45 g, 76%).

[0463] Synthesis of poly[HPMA-co-MA-GGFG-N-(6-aminohexyl)carbamoyl fumagillol] and general procedure B

[0464] [ka]

[0465] N 2 A solution of poly(HPMA-co-MA-GG-ONp) (0.68 g) in anhydrous DMF (12 mL) was diluted with H-Phe-Gly-NH(CH) in anhydrous DMF (5 mL) at 0 °C. 2 ) 6NHCO-fumagillol (0.45 g) was added followed by diisopropylethylamine (DIPEA) (0.25 mL). The reaction mixture was allowed to warm to room temperature and N 2 After stirring overnight, 3-amino-1-propanol (0.032 g) was added. The mixture was allowed to stir for an additional hour. The solvent was removed under reduced pressure and the resulting residue was dissolved in 300 mL of distilled water and extracted with EtOAc (4 times). Saturated aqueous NaCl solution (50 mL) was used to facilitate phase separation. Traces of EtOAc were removed from the polymer solution by stirring under a stream of nitrogen gas. The polymer solution was filtered through a vacuum cap filter (pH=5.56), concentrated to 30 mL by TFF in a 10K capsule, and washed with water (700 mL) by TFF. The polymer was then lyophilized to give the desired polymer conjugate poly[HPMA-co-MA-GGFG-N-(6-aminohexyl)carbamoylfumagillol] as a light pink foam (0.685 g). The spiroepoxide content was measured by reaction with 2-mercaptopyrimidine and determined to be 0.4 mmol / g.

[0466] Synthesis of poly[HPMA-co-MA-GGLG-N-(6-aminohexyl)carbamoyl fumagillol]

[0467] [ka]

[0468] The dipeptide H-Leu-Gly-NH(CH) was synthesized using standard techniques. 2 ) 6 NHCO-Fum was prepared and coupled to poly(HPMA-co-MA-GG-ONp) using general procedure B.

[0469] Synthesis of poly[HPMA-co-MA-GGVG-N-(6-aminohexyl)carbamoyl fumagillol]

[0470] [ka]

[0471] The dipeptide H-Val-Gly-NH(CH) was synthesized using standard techniques. 2 ) 6 NHCO-Fum was prepared and coupled to poly(HPMA-co-MA-GG-ONp) using general procedure B.

[0472] Synthesis of poly[HPMA-co-MA-GGGG-N-(6-aminohexyl)carbamoyl fumagillol]:

[0473] [ka]

[0474] The dipeptide H-Gly-Gly-NH(CH 2 ) 6 NHCO-Fum was prepared and coupled to poly(HPMA-co-MA-GG-ONp) using general procedure B.

[0475] Synthesis of poly[HPMA-co-MA-GFLG-N-(cis-4-aminocyclohexyl)carbamoyl fumagillol] via poly[HPMA-co-MA-GFLG-N-(cis-4-aminocyclohexylamine-HCl)]:

[0476] [ka]

[0477] General procedure C was followed using ds-1,4-diaminocyclohexane (0.914 g) and poly(HPMA-co-MA-GFLG-ONp) (1.5 g) to give poly[HPMA-co-MA-GFLG-N-(ds-4-aminocyclohexylamine-HCl)] as an off-white solid (1.08 g).

[0478] General procedure F was followed using poly[HPMA-co-MA-GFLG-N-(cis-4-aminocyclohexylamine-HCl)] (0.98 g), p-nitrophenyl fumagyl-6-yl carbonate (0.465 g), and DIEA (0.268 g) in 16 mL DMF. The solvent was evaporated and the solution was diluted with water. The aqueous phase (500 mL total) was extracted with ethyl acetate (80 mL total) and purified by TFF with an additional 350 mL water. The retentate was diluted with water, extracted with ethyl acetate, and lyophilized to give poly[HPMA-co-MA-GFLG-N-(cis-4-aminocyclohexyl)carbamoyl fumagillol] as a light pink solid (0.79 g).

[0479] 1 H NMR (DMSO-d6): δ 7.90-8.35 (m, 4H, amide-NH), 7.0-7.70 (m, 25H, phenylalanine and amide-NH), 5.26 (m, H-5-Fum), 5.18 (bt, alkene-Fum), 4.60-4.90 (m, 14H), 4.50-4.60 (m, 1H, α proton of phenylalanine), 4.10-4.30 (m, 1H, alpha proton of leucine), 3.40-3.80 (m, 21H), 3.26 (s, 3H, OMe-Fum), 2.80-3.10 (m, 31H), 2.17 (m, 2H, allyl-Fum), 0.37-2.0 [m, 166H {1.69 (s, 3H, Fum-Me), 1.59 (s, 3H, Fum-Me), 1.07 (s, 3H, Fum-Me)}].

[0480] Poly(HPMA-co-MA-GFLG-NHCH 2 CH 2 NH 2 HCl) and reaction of poly(HPMA-co-MA-GFLG-ONp) with diamines as described in General Procedure C:

[0481] [ka]

[0482] A solution of ethylenediamine (0.33 g, 5.49 mmol) in water (20 mL), pH 11.7, was adjusted to pH 9.1 by the addition of 37% aqueous HCl (17-18 drops). The solution was cooled in an ice bath and poly(HPMA-co-MA-GFLG-ONp) (1.03 g) in DMF (6 mL) was added dropwise over 20 min, maintaining the temperature below 4 °C. The solution was stirred at 4 °C for 20 min and at room temperature for 50 min to give a lemon yellow solution, pH 8.1. The solution was evaporated at 40 °C. H 2 HO (10 mL x 3) was added and evaporated. The product was diluted with water (60 mL) and the solution was adjusted to pH 8.0 with NaOH. The solution was filtered through a VacuCap filter and purified by TFF as follows. First, the polymer solution was washed with 25 mM NaCl solution (800 mL) to remove p-nitrophenol. The solution was washed with water (400 mL) and then adjusted to pH 4 with 0.1 M HCl. The TFF retentate was collected and the filter was washed with 10 mL water x 2. The retentate and washings were combined to give the polymer solution, which was lyophilized to give the compound poly(HPMA-co-MA-GFLG-NHCH 2 CH 2 NH 2 HCl) was isolated as a pale yellow solid (0.71 g, 72%).

[0483] Synthesis of N-[(2R)1-hydroxy-2-methylbutan-2-yl]carbamoyl fumagillol and general procedure D:

[0484] [ka]

[0485] A solution of p-nitrophenyl fumagyl-6-yl carbonate (400 mg, 0.89 mmol) and (R)-2-amino-3-methyl-1-butanol (280 mg, 2.71 mmol) was stirred in ethanol (10 mL) at room temperature for 12 h. The yellow solution was concentrated and the residue was purified by flash chromatography (methanol / methylene chloride) to give N-[(2R)1-hydroxy-2-methylbutan-2-yl]carbamoyl fumagillol (340 mg, 0.83 mmol) as a colorless oil.

[0486] Synthesis of N-(6-hydroxyhexyl)carbamoyl fumagillol:

[0487] [ka]

[0488] General procedure D was followed using p-nitrophenyl fumagyl-6-yl carbonate (150 mg) and 6-aminohexanol (48 mg) in ethanol (10 mL). The product was isolated as a colorless oil (110 mg, 78%).

[0489] Synthesis of N-[1-(hydroxymethyl)cyclopentyl]carbamoyl fumagillol:

[0490] [ka]

[0491] General procedure D was followed using p-nitrophenyl fumagyl-6-yl carbonate (100 mg) and cycloleucinol (52 mg) in ethanol (3 mL) and THF (1 mL) to give N-[1-(hydroxymethyl)cyclopentyl]carbamoyl fumagillol as an oil (50 mg).

[0492] Synthesis of N-(1-hydroxy-2-methylpropan-2-yl)carbamoyl fumagillol:

[0493] [ka]

[0494] General procedure D was followed using p-nitrophenyl fumagyl-6-yl carbonate (100 mg) and 2-amino-2-methylpropanol (40 mg) in ethanol (3 mL) and THF (2 mL) to give N-(1-hydroxy-2-methylpropan-2-yl)carbamoyl fumagillol as an oil (37 mg).

[0495] Synthesis of fumagyl-6-yl (2S)-2-(hydroxymethyl)pyrrolidine-1-carboxylate:

[0496] [ka]

[0497] General procedure D was followed. S-Prolinol (68 mg, 0.67 mmol) was reacted with p-nitrophenyl fumagyl-6-yl carbonate (150 mg, 0.335 mmol) in ethanol (4 mL). The product was purified by flash chromatography (methanol / methylene chloride) to give fumagyl-6-yl (2S)-2-(hydroxymethyl)pyrrolidine-1-carboxylate as a white foam (81 mg, 63%).

[0498] Synthesis of N-(6-aminohexyl)carbamoyl fumagillol:

[0499] [ka]

[0500] A solution of 1,6-diaminohexane (0.13 g) in methanol (8 mL) was cooled to 0° C. and p-nitrophenyl fumagyl-6-yl carbonate (0.13 g) in methanol (2 mL) was added dropwise. The solvent was reduced to 2 mL by rotary evaporation. Ethyl acetate was added and the organic phase was washed with water, 0.1 N NaOH, water, brine and dried over sodium sulfate. The solvent was evaporated and the residue was dissolved in ethanol (15 mL). DL-Tartaric acid (16 mg) was added and the solution was stored overnight and then evaporated to approximately 0.5 mL. Ether was added, forming a white solid. The solid was collected by filtration, washed with ether and dried to give the tartrate salt of N-(6-aminohexyl)carbamoyl fumagillol (74 mg).

[0501] Synthesis of fumagyl-6-yl[trans-(4-aminocyclohexyl)]carbamate:

[0502] [ka]

[0503] A solution of trans-1,4-diaminocyclohexane (1.3 g) in methanol (80 ml) at 0-5°C was added methanol (20 ml) and CH 2 Cl 2 A solution of fumagyl-6-yl 4-nitrophenyl carbonate (1.0 g) in 20 ml of ethyl acetate was added over 30 min, followed by stirring for 30 min. After concentration to 20 ml on a rotary evaporator and dilution with ethyl acetate (75 ml), the organic layer was washed with water (30 ml), 0.1 N NaOH (30 ml), water, and brine (30 ml), dried (MgSO 4) and concentrated under reduced pressure to give 0.78 g of a solid. This was dissolved in ethanol (80 ml) and DL-tartaric acid (127 mg) was added. After 1 hour a solution formed which was left overnight and then concentrated under reduced pressure to remove essentially all the ethanol. MTBE (100 ml) was added, concentrated and MTBE (30 ml) was added. The solid was collected by filtration, washed with MTBE (2 x 10 ml) and dried in vacuo to give fumagyl-6-yl[trans-(4-aminocyclohexyl)]carbamate hemitartrate (0.73 g), m.p. 180-185°C.

[0504] Poly[HPMA-co-MA-GFLG-NH(CH 2 ) 6 NH 2 Synthesis of -HCl: General procedure C was followed using 1,6-diaminohexane (621 mg, 5.36 mmol) and poly(HPMA-co-MA-GFLG-ONp) (1.0 g). The crude product was purified by TFF (5K) with aqueous NaCl (25 mM), then acidified to pH 4.0 with 0.1 M HCl and further purified by TFF with water to give poly[HPMA-co-MA-GFLG-NH(CH 2 ) 6 NH 2 -HCl] was obtained as an off-white solid (860 mg).

[0505] Synthesis of p-nitrophenyl N-[(2R)1-hydroxy-2-methylbutan-2-yl]carbamoyl fumagyl-6-yl carbonate and general procedure E:

[0506] [ka]

[0507] N 2To a solution of alcohol N-[(2R)1-hydroxy-2-methylbutan-2-yl]carbamoyl fumagillol (1.11 g) in methylene chloride at 0° C. was added DMAP (660 mg, 5.40 mmol) and p-nitrophenyl chloroformate (810 mg) in portions. The reaction mixture was stirred at 0° C. for 1 h. The solvent was evaporated and the resulting residue was dissolved in EtOAc, washed with water, brine and dried (Na 2 SO 4 Evaporation of the EtOAc gave the crude product which was purified by flash chromatography (silica, elution with 100% hexanes, then 2-30% EtOAc). Fractions containing pure product were combined and evaporated to isolate N-[(2R)1-(p-nitrophenolcarbonylhydroxy-2-methylbutan-2-yl]carbamoyl fumagillol (1.25 g, 80%) as a white solid.

[0508] Synthesis of N-[1-(p-nitrophenoxycarbonylhydroxymethyl)-2-methylpropan-2-yl]carbamoyl fumagillol:

[0509] [ka]

[0510] According to the general procedure E, dimethyl alcohol (60 mg), p-nitrophenyl fumagyl-6-yl carbonate (46 mg), and DMAP (37 mg) were reacted in methylene chloride (8 mL). The reaction mixture was diluted with ethyl acetate and washed with water (3 times) and then with brine. The organic phase was dried (Na 2 SO 4 ), and evaporated to give a yellow foam (87 mg) which was used without further purification.

[0511] Synthesis of N-[1-(p-nitrophenoxycarbonylhydroxymethyl)cyclopentyl]carbamoyl fumagillol:

[0512] [ka]

[0513] According to general procedure E, N-[1-(hydroxymethyl)cyclopentyl]carbamoyl fumagillol (product of Example 14, 74 mg), p-nitrophenyl chloroformate (53 mg), and DMAP (43 mg) were reacted in methylene chloride (5 mL). After extractive workup, N-[1-(p-nitrophenoxycarbonylhydroxymethyl)cyclopentyl]carbamoyl fumagillol (100 mg) was used without further purification.

[0514] Poly[HPMA-co-MA-GFLG-NH(CH 2 ) 6 Synthesis of NH-carbamoyl-[1-hydroxy-3-methylbutan-2-yl]carbamoyl fumagillol] and general procedure F:

[0515] [ka]

[0516] DIEA (0.11 g) was added dropwise to a solution of polymer (400 mg) and p-nitrophenyl N-[(2R)1-hydroxy-3-methylbutan-2-yl]carbamoyl fumagyl-6-yl carbonate (240 mg) in DMF (8 mL) at 0° C. The solution was stirred at 0° C. for 1 h and allowed to warm to room temperature. After 3 days, the solvent was evaporated and water (80 mL) was added. The aqueous phase was extracted with ethyl acetate (500 mL total) until no starting carbonate was detectable by mass spectrometry. The aqueous phase was purified by TFF (10K) and the retentate was lyophilized to give the conjugate as a white solid (380 mg, 77%).

[0517] 1H NMR (DMSO-d6): δ 8.25 (bs, 2H, amide-NH), 8.0 (bs, 1H, amide-NH), 7.70 (bs, 2H, amide-NH), 7.10-7.30 (m, 15H, phenylalanine and amide-NH), 7.10 (bt, 1H, NH-Fum), 6.92 (bd, 1H, NH-Fum), 5.26 (m, H-5-Fum), 5.18 (bt, alkene-Fum), 4.50-4.80 (m, 1H, α-protons of phenylalanine), 4.0-4.21 (m, 1H, α-proton of leucine), 3.50-3.84(m, 19H), 3.29(s, 3H, OMe-Fum), 2.80-3.10(m, 28H), 2.51(d, 1H, J = 4.4 Hz, H-2-Fum), 2.19(m, 2H, allyl-Fum), 0.82-1.92[m, 131H{1.84(m, 2H, Fum), 1.72(s, 3H, Fum-Me), 1.60(s, 3H, Fum-Me), 1.09(s, 3H, Fum-Me), 0.84(dd, 6H, Fum-isopropyl}].

[0518] Synthesis of poly[HPMA-co-MA-GFLG-N-(2-aminoethyl)carbamoyl fumagillol]:

[0519] [ka]

[0520] DMF (10mL) medium poly(HPMA-co-MA-GFLG-NHCH 2 CH 2 NH 2 -HCl) (200 mg), p-nitrophenyl fumagyl-6-yl carbonate (100 mg), and DIEA (57 mg). The product was purified by TFF (10K) in water and lyophilized to give the conjugate as a pale yellow solid (160 mg).

[0521] Synthesis of poly[HPMA-co-MA-GFLG-N(Me)-(2-methylaminoethyl)carbamoyl fumagillol]:

[0522] [ka]

[0523] DMF (5mL) medium poly(HPMA-co-MA-GFLG-N(Me)CH 2 CH 2 General procedure F was followed using NHMe-HCl) (200 mg), p-nitrophenyl fumagyl-6-yl carbonate (100 mg), and DIEA (57 mg). The product was purified using TFF (10K) with water and lyophilized to give the conjugate as an off-white solid (180 mg).

[0524] Synthesis of poly(HPMA-co-MA-GFLG-N-(2-aminoethyl)carbamoyldihydrofumagillol:

[0525] [ka]

[0526] DMF (10mL) medium poly(HPMA-co-MA-GFLG-NHCH 2 CH 2 NH 2 General procedure F was followed using p-nitrophenyl dihydrofumagyl-6-yl carbonate (200 mg), p-nitrophenyl dihydrofumagyl-6-yl carbonate (200 mg), and DIEA (57 mg). The product was purified by TFF (10K) with water (150 mL) and lyophilized to give poly(HPMA-co-MA-GFLG-N-(2-aminoethyl)carbamoyldihydrofumagillol) as a pale yellow solid (160 mg).

[0527] Synthesis of poly[HPMA-co-MA-GFLG-N-(3-aminopropyl)carbamoyl fumagillol]:

[0528] [ka]

[0529] DMF (6mL) medium poly(HPMA-co-MA-GFLG-NHCH 2 CH 2 CH 2 NH 2 General procedure F was followed using p-nitrophenyl fumagyl-6-yl carbonate (220 mg), p-nitrophenyl fumagyl-6-yl carbonate (110 mg), and DIEA (63 mg). The solvent was evaporated and the resulting solution was diluted with water. The aqueous phase was extracted with ethyl acetate and purified by TFF using 350 mL of water. The retentate was lyophilized to give poly[HPMA-co-MA-GFLG-N-(3-aminopropyl)carbamoyl fumagillol] as a light pink powder (200 mg).

[0530] Synthesis of poly[HPMA-co-MA-GFLG-N-(6-aminohexyl)carbamoyl fumagillol]:

[0531] [ka]

[0532] General procedure F was followed using poly[HPMA-co-MA-GFLG-N-(trans-4-aminocyclohexylamine-HCl)] (1.0 g), p-nitrophenyl fumagyl-6-yl carbonate (0.48 g), and DIEA (0.27 g) in DMF (25 mL). The solvent was evaporated and the solution was diluted with water. The aqueous phase (300 mL) was extracted with ethyl acetate (total 700 mL) and purified by TFF using an additional 350 mL of water. The retentate was lyophilized to give poly[HPMA-co-MA-GFLG-N-(4-aminocyclohexyl)carbamoyl fumagillol] as a light pink solid (0.9 g).

[0533] 1H NMR (DMSO-d6): δ 8.10-8.35 (m, 3H, amide-NH), 7.90-8.10 (m, amide-NH), 7.05-7.32 (m, 22H, amide-NH), 5.27 (m, H-5-Fum), 5.18 (bt, alkene-Fum), 4.60-4.90 (m, 14H), 4.50-4.60 (m, 1H, alpha protons of phenylalanine), 4.10-4.30 (m, 1H, leucine α-proton of Me), 3.40-3.80 (m, 21H), 3.27 (s, 3H, OMe-Fum), 2.80-3.20 (m, 33H), 2.56 (d, 1H, H=3.90Hz, H-2-Fum), 2.18 (m, 2H, allyl-Fum), 0.37-2.0 [m, 147H{1.70 (s, 3H, Fum-Me), 1.60 (s, 3H, Fum-Me), 1.07 (s, 3H, Fum-Me)}].

[0534] Synthesis of poly[HPMA-co-MA-GFLG-N-(trans-4-aminocyclohexyl)carbamoyl fumagillol]:

[0535] [ka]

[0536] General procedure F was followed using poly[HPMA-co-MA-GFLG-N-(trans-4-aminocyclohexylamine-HCl)] (1.0 g), p-nitrophenyl fumagyl-6-yl carbonate (0.48 g), and DIEA (0.27 g) in 25 mL of DMF. The solvent was evaporated and the solution was diluted with water. The aqueous phase (300 mL) was extracted with ethyl acetate (700 mL total) and purified by TFF using an additional 350 mL of water. The retentate was lyophilized to give poly[HPMA-co-MA-GFLG-N-(3-aminohexyl)carbamoyl fumagillol] as a light pink solid (0.9 g).

[0537] 1H NMR (DMSO-d6): δ 7.90-8.35 (m, 4H, amide-NH), 7.0-7.70 (m, 25H, phenylalanine and amide-NH), 5.26 (m, H-5-Fum), 5.18 (bt, alkene-Fum), 4.60-4.90 (m, 14H), 4.50-4.60 (m, 1H, α proton of phenylalanine), 4.10-4.30 (m, 1H, alpha proton of leucine), 3.40-3.80 (m, 21H), 3.26 (s, 3H, OMe-Fum), 2.80-3.10 (m, 31H), 2.17 (m, 2H, allyl-Fum), 0.37-2.0 [m, 166H {1.69 (s, 3H, Fum-Me), 1.59 (s, 3H, Fum-Me), 1.07 (s, 3H, Fum-Me)}].

[0538] Synthesis of poly[HPMA-co-MA-GFLG-N-[2-(4-aminophenyl)ethyl]carbamoyl fumagillol]:

[0539] [ka]

[0540] To a suspension of poly[HPMA-co-MA-GFLG-OH] (200 mg), N-[2-(4-aminophenyl)ethyl]carbamoyl fumagillol] (100 mg), and DIEA (75 mg) in DMF (6 mL) at 0° C., EDCI (44 mg total) was added portionwise. The solution was allowed to warm to room temperature and stirred overnight. The solvent was evaporated, the residue was suspended in water, and the suspension was extracted with EtOAc (7 times, 250 mL total). The aqueous phase was purified by TFF (10K) with water (350 mL). The retentate was lyophilized to give the polymer as a white fluffy solid (170 mg).

[0541] Synthesis of poly[HPMA-co-MA-GFLG-NH-2-[(2-(2-aminoethoxy)ethoxy)ethyl]carbamoyl fumagillol]:

[0542] [ka]

[0543] To a solution of 2,2'-(ethylenedioxy)bis(ethylamine) (0.79 g, 5.34 mmol) in distilled water (20 mL) at 0 °C (pH = 11.56), concentrated HCl was added until the pH of the solution was 9.01 (measured with a pH meter). DMF (6 mL) and H 2 Poly(HPMA-co-MA-GFLG-ONp) (1.0 g, 0.534 mmol) in O (10 mL) was added dropwise to the amine-containing solution over 15 min, and the reaction mixture was stirred at 0° C. for 15 min. The reaction mixture was then allowed to warm to room temperature and stirred for 2 h. The pH of the solution was measured to be 8.15. The reaction mixture was diluted with distilled water (300 mL), filtered through a VacuCap filter, and the reaction flask was washed with water (100 mL). The polymer solution was concentrated to 40 mL by TFF (10K), washed with 25 mM NaCl (800 mL) to remove p-nitrophenol, then the pH was adjusted to 4 with 0.1 M HCl, and washed with water (400 mL). Upon lyophilization of the pure polymer solution, poly[HPMA- co -MA-GFLG-NH-2-[2-(2-aminoethoxy)ethoxy]ethylamine-HCl] was isolated as a pink solid (800 mg, 78%).

[0544] N 2DIEA (57 mg, 0.416 mmol) was added to a mixture of p-nitrophenyl fumagyl-6-yl carbonate (93 mg, 0.208 mmol) and poly[HPMA-co-MA-GFLG-N-2-[(2-(2-aminoethoxy)]ethoxy)ethylamine-HCl] (200 mg, 0.104 mmol) in anhydrous DMF (5 mL) at 0 °C. The reaction mixture was allowed to warm to room temperature and stirred for 12 h. The solvent was removed under reduced pressure and the resulting residue was suspended in water (30 mL) and extracted with EtOAc (aqueous and organic phases from the emulsion formed were separated using centrifugation) to remove excess p-nitrophenyl fumagyl-6-yl carbonate and p-nitrophenol. Nitrogen was passed through the aqueous solution to remove traces of EtOAc and the solution was purified by washing with water (150 mL) using TFF (5K) to remove DIEA hydrochloride. The polymer solution was lyophilized to give the desired polymer conjugate poly[HPMA-co-MA-GFLG-N-2-[2-(2-aminoethoxy)ethoxyethyl]carbamoyl fumagillol] (220 mg, 95%) as an off-white solid.

[0545] Synthesis of poly[HPMA-co-MA-GFLG-NH-(6-aminodecyl)carbamoyl fumagillol]:

[0546] [ka]

[0547] N 2DIEA (83 mg, 0.64 mmol) was added to a mixture of p-nitrophenyl fumagyl-6-yl carbonate (300 mg, 0.67 mmol) and poly[HPMA-co-MA-GFLG-N-10-[decylamine-HCl] (300 mg, 0.15 mmol; prepared as in Example 33 except that 1,10-diaminodecane was used as the amine) in anhydrous DMF (6 mL) at <0°C. The reaction mixture was allowed to warm to room temperature and stirred for 12 h. The solvent was removed under reduced pressure and the resulting residue was suspended in water (30 mL) and extracted with EtOAc (aqueous and organic phases from the emulsion formed were separated using centrifugation) to remove excess p-nitrophenyl fumagyl-6-yl carbonate and p-nitrophenol. Nitrogen was passed through the aqueous solution to remove traces of EtOAc. The crude aqueous solution was purified by washing with water (150 mL) using TFF (10K) to remove DIEA hydrochloride. The polymer solution was lyophilized to give the desired polymer conjugate poly[HPMA-co-MA-GFLG-NH-(10-aminodecyl)carbamoyl fumagillol] (300 mg, 87%) as an off-white solid.

[0548] Synthesis of N-(2-acetamidoethyl)carbamoyl fumagillol:

[0549] [ka]

[0550] To a solution of p-nitrophenyl fumagyl-6-yl carbonate (200 mg) in ethanol (5 mL) at 0° C. was added N-(2-aminoethyl)acetamide (0.132 mL). The solution was stirred at 0° C. for 1 h and at room temperature overnight. The reaction was diluted with ethyl acetate and washed with water. The aqueous phase was back-extracted with ethyl acetate and the combined organic phases were dried (MgSO 4 The crude product was purified by flash chromatography. The product was a yellow solid (120 mg).

[0551] Synthesis of the following compound:

[0552] [ka]

[0553] Poly(HPMA-co-MA-GFLG-NHCH) in DMF (6 mL) at 0 °C 2 CH 2 NH 2 To a solution of N-(5-carboxypentyl)carbamoyl fumagillol (96 mg) was added DIEA (104 mg) followed by N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (42 mg). The solution was allowed to warm to room temperature and stirred overnight. The solvent was evaporated and the residue was dissolved in water (50 mL) and extracted with ethyl acetate (200 mL). The aqueous phase was purified by TFF using water (450 mL). The retentate was lyophilized to give the polymer (200 mg) as a pale yellow solid.

[0554] Synthesis of the following compound:

[0555] [ka]

[0556] Poly[HPMA-co-MA-GFLG-N(CH 2 ) 6 H 2 To a solution of 2-carboxyethylcarbamoyl fumagillol (91 mg), DIEA (118 mg) was added followed by N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (88 mg). The solution was allowed to warm to room temperature and stirred overnight. The solvent was evaporated and the residue was dissolved in water (50 mL) and extracted with ethyl acetate (200 mL). The aqueous phase was purified by TFF (10K) using water (1 L). The retentate was lyophilized to give the polymer (170 mg) as a pale yellow solid.

[0557] Synthesis of the following compound:

[0558] [ka]

[0559] Poly(HPMA-co-MA-GFLG-NHCH) was dissolved in DMF (6 mL) with DIEA (63 mg). 2 CH 2 CH 2 NH 2 General procedure F was followed using 1,2-dichloromethane (HCl) (220 mg) and carbonate (Example 24, 100 mg). The reaction was extracted with ethyl acetate. After purification by aqueous TFF (10K) followed by lyophilization, the product was isolated as a pale pink powder (140 mg).

[0560] BocNHCH 2 CH 2 N(Me)CH 2 C(O)NHC(O) 2 -Fumagyl-6-yl (alkylation of N-BOC, N'-methylethylenediamine with chloroacetylcarbamoyl fumagillol):

[0561] [ka]

[0562] A solution of ΤΝΡ-470 (0.2 g) and DIEA (0.105 g) in DMF (3 mL) was cooled to 0 °C. A solution of tert-butyl N-[2-(methylamino)ethyl]carbamate (0.105 g) in DMF (3 mL) was added and the mixture was stirred at 0 °C for 3 h and then overnight. The reaction was diluted with ethyl acetate and extracted with water. The aqueous phase was back-extracted with ethyl acetate and the combined organic phases were extracted with brine, dried (MgSO 4 ) and evaporated to give an oil. The product fractions were purified by silica gel chromatography (methanol / methylene chloride) and evaporated to give BocNHCH 2 CH 2 N(Me)CH 2 C(O)NHC(O) 2-Fumagyl-6-yl was obtained as a white foam (0.16 g, 60%).

[0563] Reaction of tert-butyl N-[2-aminoethyl]carbamate with chloroacetylcarbamoyl fumagillol:

[0564] [ka]

[0565] A 30 uL aliquot of a 1 M solution of Boc-ethylenediamine in DMF was added to DMF (270 uL). The solution was cooled to 0° C. and a solution of TNP-470 (48 mg) in DMF (600 uL) was added dropwise over 2 min. The reaction was monitored by LC / MS. The maximum amount of desired alkylation product observed was 34%. Carbamoyl fumagillol was also produced. The ratio of desired product to carbamoyl fumagillol was 1.0 to 0.4. Attempts to isolate the desired product resulted in the isolation of hydantoin and fumagillol. Thus, the desired product could not be isolated due to the rate of decomposition. Thus, TNP-470 could not be alkylated according to the method described.

[0566] In vivo study of DIO C57B16 mice - Body weight change, food intake, and body composition Thirteen-week-old C57B16 male mice (N=6) with an average body weight of 34 g were fed ad libitum with TD.06414, a high-fat diet (Harlan diet) with 60% of Kcal from fat. On study day 1, animals were randomly divided into groups such that the average body weight of each group was 33.9 g. Mice were treated with either phosphate-buffered saline (vehicle), TNP-470, or Compound 16 (subcutaneously in the back). Treatment continued for 31 days at the doses and schedules shown in the table below. Animals were weighed every other day. Food intake was measured once a week. On day 33, macroscopic observations were obtained to determine body composition.

[0567] FIG. 1 compares weight loss in obese DIO mice following treatment with a fumagillol-conjugate compound of the invention (compound 16) or TNP-470 (a synthetic fumagillin analog) at different doses / regimens as listed in Table 2.

[0568] [Table 4]

[0569] The results in FIG. 1 show that the vehicle control group had a weight gain of 16% and the administration of compound 16 in a q4d dosing regimen resulted in a weight loss of 19%. The administration of compound 16 provides both therapeutic and prophylactic effects. Specifically, compound 16 induces or promotes weight loss and prevents weight gain. Compound 16 is superior to TNP-470 in the extent of weight loss. Compound 16 is superior to TNP-470 in that the dose of fumagillol is reduced.

[0570] Table 3 compares body fat composition in obese DIO mice after treatment with Compound 16 or TNP-470 at different doses / regimens as described herein. Analysis was performed macroscopically on day 33. Total fat as a percentage of body weight in the vehicle group was 13.2%, while total mortality in the Compound 16-treated group was 8.2% (1 mg / kg, qod) or 5.6% (6 mg / kg, q4d).

[0571] [Table 5]

[0572] Figure 2 compares the average daily food intake in obese DIO mice after treatment with compound 16 or TNP-470 at different doses / regimens. The results in Figure 2 show reduced food intake after treatment with compound 16, which produces a greater reduction in food intake than TNP-470.

[0573] Figure 3 compares body composition (fat vs. body weight) in obese DIO mice after treatment with Compound 16 or TNP-470 at different doses / regimens. The results in Figure 3 show that reduced body weight directly correlates with reduced fat.

[0574] In vivo study of DIO C57B16 mice - body weight change, food intake, glucose tolerance, and body composition dose response Fifteen week old male C57B16 mice (N=6) with an average body weight of 42g were fed ad libitum with TD.06414, a high fat diet (Harlan diet) with 60% of Kcal composed of fat. On the first day of the study, animals were treated with either phosphate buffered saline (vehicle) or Compound 16 at different doses (subcutaneously in the back). Treatment continued for 29 days at the doses and schedules shown in the table below. Animals were weighed every other day. Food intake was measured once a week. On the 24th day (mice had been most recently treated with Compound 16 on the 21st day), an overnight fasting intraperitoneal (IP) glucose tolerance test (GTT) was performed on the vehicle and the four Compound 16-treated groups. Each animal was weighed and a baseline fasting glucose measurement was collected. Each animal was given a dose of 1 gram per kilo of glucose as a 25% solution by intraperitoneal injection. Blood glucose levels were measured 15, 30, 60, 90, and 120 min after intraperitoneal glucose administration (via tail vein blood samples using the AlphaTRAK blood glucose monitoring system (includes glucose meter and test strips) commercially available from Abbott Laboratories (North Chicago, Illinois, USA). The AlphaTRAK instrument displays results ranging from 20 to 750 mg / dL (1.1 to 41.7 mmol / L). On day 32 (mice were most recently dosed on day 29), animals were fasted for 3 h, weighed, blood was collected by cardiac puncture, and macroscopic findings were obtained to determine body composition. Blood analysis was performed by Idexx laboratories. Blood glucose was 278, 290, 265, 259, and 227 mg / dL for doses 0, 0.2, 0.6, 2.0, and 6.0, respectively. Blood urea nitrogen (BUN) was 21.8, 22.0, 19.7, 15.3, and 16.5 for doses of 0, 0.2, 0.6, 2.0, and 6.0, respectively.

[0575] Table 4 shows blood glucose levels as a function of time and dose of Compound 16. Table 4 shows that higher doses of Compound 16 resulted in lower blood glucose levels, even at the lowest dose of 0.2 mg / kg, and these results are shown in Figure 4.

[0576] [Table 6]

[0577] Table 5 shows that increasing doses of Compound 16 resulted in significantly improved weight loss at doses greater than 0.2 mg / kg q4d. Table 6 shows that doses of 2 mg / kg q4d and 6 mg / kg q4d led to significant decreases in food intake compared to vehicle controls, and that food intake was dose-responsive. From days 9-29, weekly food intake for the 2 mg / kg group was 90% of vehicle, while food intake for the 6 mg / kg group was 75% of vehicle.

[0578] [Table 7]

[0579] [Table 8]

[0580] Table 7 shows that adipose tissue is lost before other tissues, with control mice having approximately 13% fat, compared to 11% and 10% fat in the 2 mg / kg q4d and 6 mg / kg q4d groups, respectively.

[0581] [Table 9]

[0582] The results in Figure 5 show that weight loss is enhanced following treatment with Compound 16 at doses of 0.6 mg / kg and above using a q4d schedule. Weight loss was dose-responsive, with greater weight loss with increasing dose.

[0583] The results in Table 8 show a decrease in cholesterol, triglycerides, HDL, LDL, and the HDL / LDL ratio associated with increasing doses of Compound 16. These results are shown in FIG.

[0584] [Table 10]

[0585] The results in Table 9 show favorable changes in alkaline phosphatase, SGPT, SGOT, and CPK associated with increasing doses of Compound 16.

[0586] [Table 11]

[0587] Working Example Fifteen week old male C57B16 mice (N=6) with an average body weight of 42g were fed ad libitum with TD.06414, a high fat diet (Harlan diet) with 60% of Kcal composed of fat. On study day 1, animals were treated with either phosphate buffered saline (vehicle) or a dose of 2mg / kg of Compound 16, 28, 29, or 30, or 6mg / kg of Compound 31 on a q4d schedule (dorsal, subcutaneous administration). Animals were weighed every other day. Figure 10 compares weight loss in obese DIO mice after 23 days of treatment with various conjugates of the invention. The results in Figure 10 show that variation in the linker alone results in varying degrees of weight loss.

[0588] Working Example Male Sprague Dawley rats (N=3) aged 9-10 weeks with an average body weight of 300 g were fed a standard rodent diet (PharmaServ lab diet 5001) ad libitum. Figure 7 - Rats were treated with either 100 mg / kg or 200 mg / kg of compound 16 (intravenous, tail vein) on days 1, 8, 15, 22, and 29. Rats were weighed periodically and blood was collected on days 10, 17, and 24. For in-life blood collection, rats were anesthetized with an inhalation mixture of 4% isoflurane and 1.5% oxygen, and then a volume of at least 1 mL of blood was collected by puncture of the retro-orbital plexus. On day 31, animals were weighed, blood was collected by cardiac puncture, and macroscopic findings were obtained to determine body composition. Clinical findings were unremarkable for albumin, albumin / globulin ratio, alkalinity, phosphatase, ALT (SGPT), AST (SGOT), bicarbonate, direct bilirubin, indirect bilirubin, total bilirubin, BUN, BUN / creatinine ratio, calcium, chloride, cholesterol, CK, creatinine, globulin, glucose, phosphorus, potassium, sodium, sodium / potassium ratio, and total protein, as compared to normal ranges and pre-dose data. Apart from weight loss and other findings reported herein, animals appeared generally normal and did not show any evidence of neurotoxicity, such as ataxia, disorientation, tremors, or convulsions. The results in Figure 7 show that compound 16 is tolerated at high doses on a q7d dosing schedule.

[0589] Working Example Male Sprague Dawley rats (N=3, average body weight 350 g) were administered a single intravenous bolus of either vehicle, Compound 1 (30 mg / kg), or Compound 16 (200 mg / kg). Blood samples were collected by saphenous vein puncture at 0, 0.25, 0.5, 1, 2, 4, 8, 24, and 48 hours later. Aliquots of each sample were diluted in methanol containing propranolol as an internal standard and analyzed by LC / MS / MS with a lower limit of quantification of 2.5 nM. The analyte in either Compound 1 or Compound 16 administration was Compound 1. The half-life of the small molecule Compound 1 was in the range of 10-15 minutes, with a maximum plasma concentration (Cmax) of approximately 15 μM and a T 0 For the polymer conjugate compound 16, the released small molecule exhibits a maximum plasma concentration of about 0.3 μM at about 3 hours and a terminal elimination half-life of 10 hours. These results are shown in FIG.

[0590] Example: In vivo study in DIO Levin rats - body weight change, food intake, body composition, schedule-dose response, leptin levels: A study was conducted to evaluate the relative efficacy of a fumagillol polymer conjugate, compound 16, and a small molecule fumagillol derivative, compound 1, and CKD-732 (also referred to as beloranib and ZGN-433). As used herein, CKD-732 has the following structure:

[0591] [ka]

[0592] The hemitartrate salt was also tested in the hemitartrate form.

[0593] Test articles were administered subcutaneously to a diet-induced obese (DIO) Levin-DS rat model on a q4d schedule. Compound 16 efficacy was also evaluated on a q7d dosing schedule. Dietary intervention (standard Chow, Labdiet 5001; 3.4 kcal / g) was included to compare with drug intervention. Male rats as young as 3 weeks of age were fed a Harlan diet TD.06414; 5.1 kcal / g pellets with 60% of calories from fat and 21% of calories from carbohydrate. Rats were randomized into groups of 3 animals with an average weight of 595 gm prior to dosing. Rats were treated with either phosphate buffered saline (vehicle), Compound 16 or Compound 1, or CKD-732 (dorsal, subcutaneous administration). Compound 16 was dissolved in the vehicle. Compound 1 in the hemitartrate form and CKD-732 in the hemitartrate form were dissolved in ethanol and then diluted with excipients. All doses were in a volume of 5.0 ml / kg. Treatment continued for 68 days at the doses and schedules shown in Table 10 below. Because the molecular weight of CKD-732 is 15% greater than that of Compound 1, CKD-732 was administered at 1.15 mg / kg, while Compound 1 was administered at 1 mg / kg for molar comparison purposes. At the first dose on Day 1, the rats were 14 weeks old. Also on Day 1, Group 2 was switched from a high-fat diet to a standard chow diet, while the other groups remained on a high-fat diet for the duration of the study.

[0594] [Table 12]

[0595] Animals were weighed every other day. Food intake was measured once a week. Blood samples were taken for evaluation of serum chemistry, including glucose and insulin, approximately once a week throughout the study. On day 48, all rats underwent a 4-hour fasting oral glucose tolerance test (OGTT). Animals were administered 8 mL / kg 25% glucose (2 g / kg) orally (per os, PO). On day 68, gross observations were obtained to determine body composition.

[0596] FIG. 11 shows the change in body weight versus test day for each group. Both Q4D and Q7D polymer conjugate groups showed a significant decrease in body weight. Treatment with compound 16 at 3 mg / kg (Q4D) or 6 mg / kg (Q7D) showed a greater weight loss than the change to standard chow. At the end of the study, compound 16 at 3 mg / kg on a Q4D schedule showed a body weight 22.1% lower than vehicle controls and 6.2% lower than rats on a standard chow diet. After about 10 weeks, treatment with compound 16 at 6 mg / kg on a Q7D schedule showed a body weight similar to treatment with a 3 mg / kg dose on a Q4D schedule. Compound 1 administered at 1 mg / kg on a Q4D schedule and CDK-732 administered at 1.15 mg / kg showed a body weight 3.9% or 3.2% lower than vehicle. Compound 1 administered at 3 mg / kg on a Q4D schedule showed a body weight 8.9% lower than vehicle. The polymer complex is approximately 1 / 6 the active fumagillol derivative by weight.

[0597] Figure 12 shows the final body weights on day 68 for all groups as a function of the average daily dose of fumagillol exposure. Both vehicle and standard diets had no fumagillol exposure. Compound 16 shows greater weight loss with significantly lower fumagillol exposure than Compound 1 or CKD-732 on the same schedule as this polymer conjugate. All groups were dosed on a Q4D schedule, except for Compound 16 at 6 mg / kg, which was dosed Q7D.

[0598] [Table 13]

[0599] FIG. 13 shows the reduction in serum insulin levels in male Levin DIO rats maintained on a 60% fat diet and administered a compound of the invention on a q4d (3 mg / kg) and q7d (6 mg / kg) schedule compared to standard dietary intervention and vehicle groups.

[0600] [Table 14]

[0601] Table 12 shows the change in fasting insulin levels for each group. All groups (except the vehicle control group) showed a decrease in insulin levels, illustrating that the compounds of the present invention lower insulin levels on a less frequent dosing schedule.

[0602] Figure 14 shows the results of oral glucose tolerance test (OGTT) on insulin levels in rats treated with the compounds of the present invention on a q4d and q7d schedule, compared with standard diet intervention and vehicle group.Standard diet intervention also resulted in lower insulin levels.Compared to vehicle, insulin levels remain reduced in the presence of abnormally high glucose levels, indicating that lower levels of insulin are required to reduce blood glucose (see Figure 15), suggesting improved / restored insulin sensitivity.

[0603] FIG. 15 shows reduced glucose levels over time with different treatments following an oral glucose challenge.

[0604] [Table 15]

[0605] FIG. 16 shows the commonly accepted index of insulin sensitivity, glucose (mM / L)×insulin (uU / ml) / 22.5, in male Levin DIO rats (Matthews et al., Diabetologia (1985) 28, 412±419; Pickavance et al., British Journal of Pharmacology (1999) 128, 1570±1576).

[0606] [Table 16]

[0607] The adipocyte hormone leptin is a known appetite suppressant. Leptin resistance (abnormally high levels independent of food intake) is known to occur in patients and animals with diet-induced obesity (Levin et al., Am J Physiol Regul Integr Comp Physiol. 2002 Oct;283(4):R941-8). Low levels of leptin have been linked to hyperphagia (Sindelar et al., 1999, Enriori et al., 2006). Food intake was measured once a week. Serum leptin levels were measured on day 29 and plotted against food intake for the week including day 29. Animals on a standard chow diet were hyperphagic and showed significantly greater food intake relative to leptin levels than did the compounds of the invention. Treatment with Compound 1 did not result in a significant decrease in leptin levels. Figure 17 shows weekly food intake in grams for each group. The standard chow group showed a significant increase in food intake after switching from a high-fat diet to a standard diet. Binge eating is known to occur in order to maintain caloric intake.

[0608] [Table 17]

[0609] Figure 18 shows the change from baseline in leptin levels in male Levin DIO rats maintained on a high fat diet and treated with the complex of the present invention or standard chow intervention. A dose-dependent response was observed in the change from baseline in leptin levels for compound 16.

[0610] [Table 18]

[0611] Example: In vivo study of DIO mice - body weight change, food intake, schedule - dose response Male C57B1 / 6 mice (N=9 / group) aged 21 weeks with an average body weight of 46.8 g were fed a high-fat diet with 60% of calories from fat ad libitum. Animals were dosed according to the schedule in Table 17 below.

[0612] [Table 19]

[0613] Compound dosing occurred between 9-10 am on the day of dosing. Groups 6 and 7 received a total of 17 doses. q4d groups (1, 2, 3, 4, 8, 9) received a total of 9 doses. q8d group (5) received a total of 5 drug doses. Body weight and food intake were measured every other day. Blood glucose was measured in the fed state at 9:00 am on days -7, 0, 7, 14, 21, and 28 (blood glucose was measured before dosing on dosing days). Blood glucose was measured with a glucometer. An intraperitoneal glucose tolerance test (ipGTT, 6-hour fast) was performed.

[0614] The study was terminated on day 34. Liver and epididymal white adipose tissue (eWAT) were harvested, weighed, and stored at -80°C. Serum was collected and AST, ALT, ALP, CK, BUN, creatinine, calcium, potassium, sodium, chloride, total protein, albumin, total bilirubin, glucose, triglycerides, and cholesterol were determined. Insulin samples were measured using a commercially available kit.

[0615] The polymer referred to in this example is poly[HPMA-co-MA-GFLG-N-(6-aminohexyl)acetamide, a fumagillol-free polymer. The synthesis of poly[HPMA-co-MA-GFLG-N-(6-aminohexyl)acetamide is described in WO 2011 / 150022, which is incorporated herein by reference in its entirety.

[0616] Structure of poly[HPMA-co-MA-GFLG-N-(6-aminohexyl)acetamide]

[0617] [ka]

[0618] Figure 19 shows the surprising and unexpected finding that the 12 mg / kg dose on a Q8D schedule results in greater initial weight loss than the 6 mg / kg dose on a Q4D schedule. By the end of the study, the Q8D group stopped losing weight, while the 6 mg / kg group appeared to continue losing weight. Polymers without fumagillol show similar weight changes to the vehicle.

[0619] Figure 20 shows that the small molecule CKD-732 (Compound B) administered on a Q2D (QOD) schedule responded better than the same average daily dose administered on a Q4D schedule. As expected, the small molecule responds better than more frequent dosing.

[0620] Figure 21 shows the reduction in food intake with compounds of the invention. Note the initial significant reduction in food intake in the 12 mg / kg group on a Q8D schedule, followed by recovery and then a cyclical reduction-recovery pattern.

[0621] Figure 22 shows significantly decreased insulin levels during ipGTT in male C57B16 mice maintained on a high fat diet. Compounds of the present invention significantly decreased the amount of insulin excreted by β-cells in the presence of hyperglycemia, indicating decreased resistance and improved insulin sensitivity. Note that fasting insulin was also decreased in all Compound 16 group mice.

[0622] FIG. 23 shows the change in total insulin AUC in male C57B16 mice maintained on a high fat diet during the glucose challenge as a function of treatment group.

[0623] FIG. 24 shows that blood glucose was reduced over the treatment period compared to the vehicle and polymer groups.

[0624] Figure 25 shows the product of glucose (mg / dl) and insulin (μU / ml) / 405 (Akagiri et al., A Mouse Model of Metabolic Syndrome, J. Clin. Biochem. Nutr., 42, 150-157, March 2008), i.e., the HOMA-ir measurement, which is a recognized measure of insulin resistance and a predictor of cardiovascular disease (Bonora et al., Diabetes Care. 2002, 25, 1135-1141).

[0625] [Table 20]

[0626] Example Efficacy of various compounds in the DIO mouse model C57B16 male mice (N=6) were fed ad libitum with TD.06414, a high fat diet (Harlan diet) consisting of 60% of Kcal from fat. On the first day of the study, animals were randomly divided into groups such that the average weight of mice in each group was 47 g. Mice were treated with either phosphate buffered saline (vehicle) or a compound listed in Table 19 dissolved in vehicle (dorsal, subcutaneous administration). Treatment continued for 26 days at the doses and schedules shown in Table 19 below. The polymer referred to in this example is poly[HPMA-co-MA-GFLG-N-(6-aminohexyl)acetamide, a fumagillol-free polymer.

[0627] [ka]

[0628] Compound cis-16 is compound 16 in which the 1,4-diaminocyclohexane is in the cis configuration rather than the trans configuration as shown in the diagram of compound 16.

[0629] Compound aa is the reaction product of a 2KDa MW methoxy-terminated PEG amine and p-nitrophenyl fumagyl-6-yl carbonate.

[0630] [ka]

[0631] Compound bb is:

[0632] [ka]

[0633] The synthesis of poly[HPMA-co-MA-GFLG-NH-2-[(2-(2-aminoethoxy)ethoxy)ethyl]carbamoyl fumagillol] is described in WO 2011 / 150022, which is incorporated by reference in its entirety.

[0634] [Table 21]

Claims

1. A pharmaceutical composition for improving or restoring insulin sensitivity in a subject, wherein the subject is overweight or obese, and the pharmaceutical composition comprises a compound having the following formula: 【Chemistry 1】 【Chemistry 2】 A pharmaceutical composition comprising a compound selected from the group consisting of: or a pharma- ceutically acceptable salt thereof.

2. The subject has a resistance of 25 kg / m 2 ~29.9kg / m 2 30 kg / m 2 More than 35kg / m 2 or more; or 40 kg / m 2 The pharmaceutical composition of claim 1, having a BMI of 0.1 or higher.

3. 2. The pharmaceutical composition of claim 1, wherein the subject has at least one obesity-induced or obesity-related comorbidity selected from the group consisting of diabetes, non-insulin-dependent type II diabetes, impaired glucose tolerance, impaired fasting glucose, elevated plasma insulin concentration, insulin resistance syndrome, hyperlipidemia, dyslipidemia, hypertension, hyperuricemia, gout, coronary artery disease, heart disease, myocardial infarction, angina pectoris, sleep apnea, obstructive sleep apnea, Pickwickian syndrome, fatty liver, cerebral infarction, stroke, cerebral thrombosis, respiratory complications, cholelithiasis, gallbladder disease, kidney disease, gastroesophageal reflux, stress urinary incontinence, arteriosclerosis, heart disease, heart rhythm abnormalities, arrhythmia, transient ischemic attack, orthopedic disorders, osteoarthritis, osteoarthritis, lumbodynia, menstrual disorders, endocrine disorders, hormonal imbalances, and infertility.

4. 2. The pharmaceutical composition of claim 1, further comprising treating, reducing or ameliorating one or more cardiometabolic risk factors in said subject, preferably selected from plasma triglyceride levels, LDL-cholesterol levels, C-reactive protein (CRP) levels, systolic blood pressure and diastolic blood pressure.

5. The pharmaceutical composition of claim 1 , further comprising administering a second active agent.

6. 2. The pharmaceutical composition of claim 1, wherein the therapeutically effective amount is from about 0.0001 mg / kg to about 5 mg / kg of body weight / day, or from about 0.001 to about 1 mg / kg of body weight / day.

7. 2. The pharmaceutical composition of claim 1, wherein the compound is suitable for administration from about 1 to about 5 times per week, preferably once every two weeks, more preferably on a q4d dosing schedule, or most preferably on a q7d dosing schedule.

8. The pharmaceutical composition of claim 1 , wherein the compound is suitable for parenteral or subcutaneous administration.

9. The method of claim 1 , wherein the compound is provided as a pharmaceutical composition comprising the compound and a pharma- ceutically acceptable carrier.

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