Comune Latihan Di Duhelin Umat

COMUNE US009783592B2LATIHAN DI DUHELIN UMAT (12 ) United States Patent (10 ) Patent No. : US 9 ,783 , 592 B2 DiMarchi et al. (45 ) Date of Patent: * Oct . 10 , 2017

(54 ) GLUCAGON SUPERFAMILY PEPTIDES 6 , 191 , 102 B12 / 2001 DiMarchi et al . 6 ,583 , 111 B1 6 / 2003 DiMarchi et al. EXHIBITING NUCLEAR HORMONE 6 , 677 ,136 B2 1 / 2004 Marshall et al. RECEPTOR ACTIVITY 7 , 192 , 922 B2 3 / 2007 Shannon et al . 7 , 211 , 557 B2 5 / 2007 DiMarchi e (71 ) Applicant: Indiana University Research and 7 ,576 , 059 B2 8 / 2009 Jonassen et al. Technology Corporation , Indianapolis , 2003/ 0021795 Al 1 / 2003 Houston et al. 2003 /0143183 Al 7 / 2003 Knudsen et al. IN (US ) 2003 / 0204063 Al 10 / 2003 Gravel et al . 2004 / 0002468 A1 1 / 2004 Wadsworth et al. ( 72 ) Inventors : Richard D . DiMarchi, Carmel , IN 2004 / 0235710 AL 11 /2004 DeFilippis et al. (US ); Bin Yang , Bloomington , IN (US ) ; 2005 / 0070469 Al 3 / 2005 Bloom et al. Brian Finan , Bloomington , IN (US ) 2005 / 0095679 A 5 / 2005 Prescott et al . 2005 /0124550 A 6 / 2005 Peri 2005 / 0153890 A1 7 / 2005 Pan et al. ( 73 ) Assignee: Indiana University Research and 2005 / 0288248 A1 12 / 2005 Pan et al . Technology Corporation , Indianapolis , 2006 /0003417 A1 1/ 2006 Pan et al . IN (US ) 2006 / 0003935 A1 1 / 2006 Pan et al . 2006 /0171920 A1 8 / 2006 Shechter et al. @( * ) Notice : Subject to any disclaimer , the term of this 2006 /02 10534 Al 9 / 2006 Lee et al. patent is extended or adjusted under 35 2006 / 0252916 Al 11 / 2006 DiMarchi et al . 2006 / 0286129 A1 12 / 2006 Sarubbi U . S . C . 154 (b ) by 0 days. 2007 / 0042956 A1 2 /2007 Johansen et al. 2007 /0173452 A1 7 / 2007 DiMarchi et al. This patent is subject to a terminal dis 2007 / 0203058 Al 8 / 2007 Lau et al . claimer . 2007 /0287670 A1 12 /2007 Natarajan et al. 2008 /0125574 A1 5 / 2008 Sheffer et al. (21 ) Appl. No. : 14 /808 , 668 2008 / 0318837 Al 12 / 2008 Quay et al. 2009 / 0036364 A1 2 / 2009 Levy et al. 2009 / 0054305 Al 2 /2009 Schlein et al. ( 22 ) Filed : Jul . 24 , 2015 2009 /0062192 A1 3 / 2009 Christensen et al. 2009 /0074769 Al 3 /2009 Glaesner et al. (65 ) Prior Publication Data 2009 / 0137456 Al 5 / 2009 DiMarchi et al . US 2015 /0320871 A1 Nov . 12 , 2015 (Continued ) Related U . S . Application Data FOREIGN PATENT DOCUMENTS (63 ) Continuation of application No. 13/ 697 , 018 , filed as CA 2024855 3 / 1992 application No . PCT /US2011 /035913 on May 10 , ?? 0220958 5 / 1987 2011, now Pat . No. 9 , 127 , 088 . EP 0479210 4 / 1992 E 0815135 9 / 1996 EP 1695983 B1 8 / 2006 (60 ) Provisional application No . 61/ 432 , 077 , filed on Jan . 2036539 Al 3 / 2009 12 , 2011 , provisional application No. 61/ 334 , 435 , EP 2036923 A1 3 / 2009 filed on May 13 , 2010 . EP 2398483 8 / 2010 ?? 2300035 1 / 2012 (51 ) Int. Cl. WO WO 96 / 29342 9 / 1996 CO7K 147605 ( 2006 .01 ) (Continued ) A61K 38 / 26 ( 2006 .01 ) COZK 14 /72 ( 2006 . 01 ) OTHER PUBLICATIONS A61K 47 /48 ( 2006 .01 ) (52 ) U . S . CI. “ Application of Chemical Biotechnology to Optimization of Endo CPC . . . C07K 14 / 721 ( 2013 .01 ) ; A61K 38 / 26 crine Hormones, ” Carothers Lecture , Mar. 22 , 2007 . (2013 .01 ) ; A61K 47 /481 ( 2013 .01 ) ; A61K (Continued ) 47/ 48123 ( 2013 .01 ) ; A61K 47/ 48246 ( 2013. 01 ); CO7K 14 /605 (2013 . 01 ); COOK Primary Examiner — Gyan Chandra 14 /72 (2013 . 01 ) ( 74 ) Attorney , Agent, or Firm - Barnes & Thornburg (58 ) Field of Classification Search LLP CPC ...... A61K 47 / 48 ; A61K 47 / 48123 ; A61K 47 / 246 ; CO7K 14 /605 (57 ) ABSTRACT See application file for complete search history . Provided herein are glucagon superfamily peptides conju gated with NHR ligands that are capable of acting at a (56 ) References Cited nuclear hormone receptor. Also provided herein are phar U . S . PATENT DOCUMENTS maceutical compositions and kits of the conjugates of the invention . Further provided herein are methods of treating a 4 , 275 , 152 A 6 / 1981 Esders et al. disease , e . g . , a metabolic disorder, such as diabetes and 5 , 359 ,030 A 10 / 1994 Ekwuribe 5 ,510 , 459 A 4 / 1996 Smith et al. obesity , comprising administering the conjugates of the 5 ,512 , 549 A 4 / 1996 Chen et al. invention . 5 ,665 , 705 A 9 / 1997 Merrifield et al. 5 , 783 ,674 A 7 / 1998 Geysen 21 Claims, 59 Drawing Sheets US 9 , 783, 592 B2 Page 2

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Tschoep , Matthias, " Afferent Gut Hormones in the Control of PCT International Search Report for PCT/ US2010 / 059724 com Energy Balance and Metabolism ” presentation slides for the 21st pleted by the US Searching Authority on Jun . 14 , 2011 . American Peptide Symposium ( Jun . 7 - 12 , 2009, Bloomington , IN ). Perret et al. , “ Mutational analysis of the glucagon receptor : simi Un Son et al. , Positively Charged Residues at Positions 12 , 17 , and larities with the vasoactive intestinal peptide (VIP ) /pituitary 18 of Glucagon Ensure Maximum Biological Potency , J Biol. adenylate cyclase -activating peptide (PACAP ) / secretin receptors Chem ., 273 ( 17 ) : 10308 - 12 ( 1998 ) . for recognition of the ligand ' s third residue ,” J . Biochem . , 362 Unson et al. , " Glucagon antagonists : Contribution to binding and ( 2002 ) , pp . 389 - 394 . activity of the amino - terminal sequence 1 - 5 , position 12 and the Phillips et al. , “ Supramolecular Protein Engineering : Design of putative alpha -helical segment 19 -27 , " J . Biol . Chem . v264, pp . Zinc - Stapled Insulin Hexamers As a Long Acting Depot, ” J. Biol. 789 - 794 , Jan . 15 , 1989 , p . 792 , para 1 , Table 1 . Chem ., vol. 285 , No. 16 , Apr. 16 , 2010 , pp . 11755 - 11759 . Unson et al. , “ Role ofHistidine - 1 in Glucagon Action , ” Archives of Pocai et al. , “ Glucagon - Like Peptide 1 /Glucagon Receptor Dual Biochemistry and Biophysics , vol. 300, No . 2 , pp . 747 - 750 , Feb . 1 , Agonism Reverses Obesity in Mice, " Diabetes, vol. 58 , No . 10 , Oct. 1993 1 , 2009 , pp . 2258 - 2266 . Vijayalakshmi et al ., “ Comparison of Helix - Stabilizing Effects of a , Robberecht , P . et al ., “ Receptor Occupancy and Adenylate Cyclase n - dialkyl Glycines with Linear and Cycloalkyl Side Chains” , Activation in Rat Liver and Heart Membranes by 10 Glucagon Biopolymers 53 : 84 - 98 ( Jan . 21 , 2000 ) . Analogs Modified in Position 2, 3 , 4, 25 , 27 and /or 29 ,” Regulatory Walensky et al ., “ Activation of Apoptosis in Vivo by a Hydrocar Peptides , 21 ( 1988 ) , 117 - 128 . Ronacher et al. , " Ligand - selective transactivation and transrepres bon - Stapled BH3 Helix ” , Science 205 : 1466 - 1470 (Sep . 3 , 2004 ) . sion via the glucocorticoid receptor: role of cofactor interaction ,” Ward et al. , In vitro and in vivo evaluation of native glucagon and Mol. Cell Endocrinol. 299 ( 2 ) : 216 - 231, p . 220 (2009 ) . glucagon analog (MAR -D28 ) during aging : lack of cytotoxicity and Santos et al. , Cyclization - Activated Prodrugs. Synthesis , Reactivity preservation of hyperglycemic effect , J . Diabetes Sci. Technol . , and Toxicity of Dipeptide Esters of Paracetamol, Bioorganic & 4 (6 ): 1311 - 21, Nov . 1 , 2010 . Medicinal Chemistry Letters 15 : 1595 - 1598 (2005 ) . Ward , “ Fatty Acid Acylation of Peptides : Developing strategies to Sapse et al ., The Role of Sale Bridge Formation in Glucagon : An enhance medicines for treating metabolic disorders ,” Jan . 14 , 2009 . Experimental and Theoretical Study of Glucagon Analogs and Ward , B . ; Finan , B . ; Gelfanov, V . and DiMarchi , R . Exploring the Peptide Fragments of Glucagon ,Malec .Med ., 8 (5 ): 251 -62 , May 1 , N - terminalHydrophobic Faces of Glucagon and Glucagon - like 2002 . Peptide- 1, (2009 ) Proceedings of the 21st American Peptide Society Sato , H . , “ Enzymatic procedure for site - specific pegylation of 153 - 154 . proteins, ” Advanced Drug Delivery Reviews 54 , pp . 487 -504 Wibowo, Synthesis , Purification , and Biological Activity of AIB ( 2002 ) . Substituted Glucagon and GLP - 1 Peptide Analogues (2005 - 2006 ) Schafmeister et al . , “ An All -Hydrocarbon Cross -Linking System for vol. 45 , 707 = 738 , accessed httDs: // scbolarworks. iu . edu /dsnce / Enhancing the Helicity and Metaboli Stability of Peptides” , J. Am . bandle / 2022 / 326 on Jul. 17 , 2012 . Chem . Soc . 122 : 5891 -5892 (2000 ) . Wynne et al. , “ Subcutaneous Oxyntomodulin Reduces Body Weight Son et al. , “ Preparation and Structural, Biochemical, and Pharma ceutical Characterizations of Bile Acid -Modified Long - Acting in Overweight and Obese Subjects , ” Diabetes , vol. 54 , Aug . 2005 , Exendin - 4 Derivatives, ” J . Med . Chem . , ACS, vol. 52 , No. 21, Nov . pp . 2390 - 2395 . 12 , 2009 , pp . 6889 -6896 . Yang et al . , “ A Novel Approach to Resin - Based Cysteine Stigsnaes et al. , “ Characterisation and Physical Stability of Alkylation , ” American Peptide Society , 2005 . PEGylated Glucagon ,” International Journal of Pharmaceutics , Yang et al. , “ A Novel Approach to Resin - Based Cysteine 330 ( 2007 ) , pp . 87 - 98 . Alkylation , ” poster presentation to American Peptide Society , 2005 . Sturm et al. , “ Structure -Function Studies on Positions 17 , 18 , and 21 Yang et al ., “ Synthesis and Biological Assessment of Sulfonic Replacement Analogues of Glucagon : The Importance of Charged Acid - Based Glucagon Antagonists, ” American Peptide Society , Residues and Salt Bridges in Glucagon Biological Activity ," J Med 2005 . Chem . 1998 , 41 , 2693 -2700 . Yang et al ., “ Synthesis and Biological Assessment of Sulfonic Sueiras - Diaz et al. , " Structure - Activity Studies on the N - Terminal Acid - Based Glucagon Antagonists, ” poster presentation to Ameri Region of Glucagon , " J . Med . Chem . ,27 , pp . 310 - 315 , 1984 . can Peptide Society , 2005 . Supplemental European Search Report issued in connection with EP Yang et al. , Synthesis and Biological Assessment of Sulfonic Application No . 09800752 issued on Jun . 20 , 2011. Acid - Based Glucagon Antagonists , Understanding Biology Using Supplemental European Search Report issued in connection with EP Peptides , American Peptide Symposia, 9 ( Part 6 ) : 305 - 6 (2006 ) . Application No. 09767567 . 2 issued on Jun . 17 , 2011. Yang , B . and DiMarchi, R . D . (2005 ) . A Novel Approach to Resin Traylor et al. , Identification of the High Potency Glucagon Agonist based Cysteine Alkylation Peptides: Chemistry , Structure and Biol with Enhanced Biophysical Stability and Aqueous Solubility , Poster ogy , Proceedings of the XIX American Peptide Symposium , ( 88 Abstract PY 10 , pp . 505 -506 , Jun . 10 , 2005 . 89 ) . Trivedi, D . et al ., Design and synthesis of conformationally con Zakrzewska et al. , “ Induction of Obesity and Hyperleptinemia by strained glucagon analogues, J . Med . Chem ., 43 ( 9 ) : 1714 - 22 , May Central Glucocorticoid Infusion in the Rat, ” Diabetes, 1999 , 4 , 2000 ( Abstract ) . 48 (2 ): 365 - 370 . Tschoep et al ., “ A Novel Glucagon /GLP - 1 Co -Agonist Eliminates Zhang et al. , Design and synthesis of novel GLP1 analogues with Obesity in Rodents , ” American Diabetes Association Abstract No. significantly prolonged time action , Biopolymers. , 80 ( 4 ): 555 313 -OR (2009 ). ( 2005 ) . U. S . Patenatent Oct. 10 , 2017 Sheet 1 of 59 US 9 ,783 , 592 B2

29 28 27 31 27

- - -- I | TIL TIL II TL LL LL II TL TILL - IT IT- HADGSFSDEMNTIIDNLAARDFINWLIOTKITD-33 - - HSQGTFTSDYSKYLDSRRAQDFVQWLMNTKRNRNNIA-37 ILI _ - - HGEGTFTSDLSKQMEEEAVRLFIEWLKNGGPSSGAPPPS-39 - HSQGTFTSDYSKYLDSRRAQDFVQWLMNT-29 HSDGIFTDSYSRYRKQMAVKKYLAAVL-- YADAIFTNSYRKVLGQLSARKLLQDIMSR- HADGVFTSDFSKLLGOLSAKKYLESLM- HSDAVFTDNYTRLRKOMAVKKYLNSILN- HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRG- YAEGTFISDYSIAMDKIHQQDFVNWLLAQKGKKNDWKHNITQ42 HSDGTFTSELSRLREGARLQRLLQGLV- AlignmentofAminoAcidSequencesandGlucagonSuperfamilyPeptides Fig.1

PACAP-38 Exendin-4 1GLP- Glucagon Oxyntomodulin GLP-2 GHRH PHI VIP GIP Secretin atent Oct. 10 , 2017 Sheet 2 of 59 US 9 ,783 , 592 B2

Change in BodyWeight

M* OSI . A ABW(%) I

2006 5 10 15 20 25 Time (Days )

? Vehicle . ..* *. . . GLP - 1 (AIB2 , A22 , Cex , K40 ) - - GLP - 1 (AIB2 , A22 ,Cex , K40 ) /Estrogen ( 17 - ester) 40 ug /kg -day - - - - GLP - 1 (AIB2 , 422 ,Cex , K40 ) /Estrogen ( 17 - ester ) 400 ug /kg - day

Fig . 2a U. S . Patentatent Oct.. 10 10 , , 20172017 Sheet 3 of 59 US 9 ,783 , 592 B2

Blood Glucose Levels

BG(mg/dL) HO 1 7 14 7 14 7 14 7 14 Time (Days ) O Vehicle Z GLP - 1 (AIB2 , 422 ,Cex , K40 ) W GLP - 1 (AIB2 , 422 ,Cex , K40 )/ Estrogen (17 - ester ) 40 ug /kg -day Z GLP -1 (AIB2 , 422 ,Cex , K40 )/ Estrogen ( 17 - ester) 400 ug/ kg - day Fig . 2b U. S . Patentatent Oct. 10 , 2017 Sheet 4 of 59 US 9 ,783 , 592 B2

Blood Glucose Levels

ala

ono HM BG(mg/dL) TI ã

- 0 7 14 0 7 14 0 7 14 0 7 14 Time (Days )

O Vehicle VA GLP - 1 (AIB2 , A22 , Cex , K40 ) WA GLP - 1 (AIB2 , A22 ,Cex , K40 )/ Estrogen (17 - ester) 40 ug /kg -day Z GLP - 1 (AIB2 , 422 ,Cex , K40 )/ Estrogen ( 17 - ester ) 400 ug/ kg -day Fig . 2c atent Oct. 10 , 2017 Sheet 5 of 59 US 9 ,783 , 592 B2

Day 21 ipGTT detgodop

BG(mg/dL) hvor - RSS- S

e 30 60 90 120 Time (Mins ) A Vehicle ------GLP - 1 (AIB2 , E16 ,Cex ,K40 ) 40 ug /kg -day - - - - GLP - 1 (AIB2 , A22 ,Cex , K40 )400 ug/ kg - day - - - - GLP - 1 (AIB2 , Ala22, Cex , K40 y/ Estrogen ( 17 -ester ) 4 ug/ kg - day - R - GLP - 1 (AIB ? ,Ala22 , Cex , K40 yEstrogen (17 -ester ) 40 ug /kg -day - .. . - . GLP - 1 (AIBP , Ala22 ,Cex , K40 y Estrogen (17 -ester ) 400 ug /kg -day Fig . 3a atent Oct. 10 , 2017 Sheet 6 of 59 US 9 ,783 , 592 B2

Change in Body Weight

ABW(%) ????? ITSHKi

0 5 10 15 20 25 Time (Days )

A Vehicle ------GLP - 1 (AIB2 , E16 ,Cex , K40 ) 40 ug/ kg -day - - - - GLP - 1 (AIB2 , A22 ,Cex , K40 ) 400 ug/ kg -day - - - - - GLP - 1 (AIB2 , Ala22 ,Cex ,K40 y/ Estrogen (17 - ester) 4 ug/ kg -day - R - GLP - 1 ( AIB ? ,Ala22 , Cex , K40 yEstrogen ( 17 -ester ) 40 ug /kg -day - O - GLP - 1 (AIB ?, Ala22 ,Cex , K40 \/ Estrogen (17 -ester ) 400 ug /kg -day Fig . 3b U. S . Patent atent Oct. 10 , 2017 Sheet 7 of 59 US 9 ,783 , 592 B2

Change in Fat Mass

8cöoo AFatM

TUDay 23 O Vehicle ZA GLP - 1 (AIB2 , E16 ,Cex , K40 ) 40 ug/ kg -day Z GLP - 1 (AIB ?, A22 ,Cex , K40 ) 400 ug /kg -day W GLP - 1 (AIB2 , Ala22 , Cex ,K40 Estrogen (17 - ester) 4 ug /kg -day Z GLP - 1 (AIB2 , Ala22 , Cex , K40 yEstrogen (17 -ester ) 40 ug /kg -day N GLP 1 (AIB2 , Ala22 , Cex , K40 YEstrogen ( 17 -ester ) 400 ug/ kg - day Fig . 30 atent (Oct . 10 , 2017 Sheet 8 of 59 US 9 ,783 , 592 B2

Change in Lean Mass

«LeanMass(%) 2727 / 11 " } ??? 17|

Day 23

O Vehicle VITA GLP - 1 ( AIB2, 516, Cex , K40) 40 ??/ kg - day 22 GLP- 1 ( AIB2, A22 , Cex, K40 ) 400 ug/ kg - day LA GLP- 1 ( AIB2, Ala22 , Cex, K40 }/ Estrogen (17 - ester ) 4 ug/ kg -day EZ GLP- 1 ( AIB2, Ala22 , Cex , K40 }/ Estrogen ( 17 -ester )40 g/ kg - day A GLP 1 ( AIB2, Ala22, Cex, K40 }/ Estrogen ( 17 - ester )400 ug/ kg - day Fig . 3d U. S. Patentatent OctDet.. 1010. , 2017 Sheet 9 9 ofor 59so US 9 ,783 , 592 B2

Change in Blood Glucose

ABG(mg/dL) ???? og HI ZA

Day 14

D Vehicle Z GLP - 1 (AIB2 , E16 ,Cex ,K40 ) 40 ug /kg -day Z GLP - 1 (AIB2 , A22 ,Cex ,K40 ) 400 ug /kg -day W GLP - 1 (AIB ? ,Ala22 ,Cex ,K40 yEstrogen (17 -ester ) 4 ug/ kg -day WZ GLP - 1 (AIB2 , Ala22, Cex, K40 y/ Estrogen (17 - ester )40 ug/ kg -day N GLP 1 (AIB2 , Ala22 ,Cex , K40 y/ Estrogen ( 17 -ester ) 400 ug /kg -day Fig . 3e atent Oct. 10 , 2017 Sheet 10 of 59 US 9 ,783 ,592 B2

Blood Glucose Levels 500 t CH

BG(mg/dL) ?potu?e?i ending

o 0 15 30 60 90 120 Day 14

a Vehicle - - 0 - - GLP - 1 (AIB , E16 , Cex , K40 ) 40 ug /kg -day - - 0 - - - GLP - 1 (AIB2 , E16 ,Cex ,K40 y/ Estrogen ( 17 -ester ) 40 ug/ kg -day - - GLP - 1 (AIB2 , E16 ,Cex ,K40 YEstrogen (3 -ether ) 40 ug /kg -day GLP -1 (AIB ,E16 ,Cex , K40 ) 400 ug/ kg -day . GLP - 1 (AIB2 , E16 ,Cex , K40 YEstrogen ( 17 -ester ) 400 ug /kg -day 600+ GLP - 1 (AIB2 , E16 ,Cex , K40 YEstrogen ( 3 - ether) 400 ug/ kg -day Fig . 4a U . S . Patent Oct. 10 , 2017 Sheet 11 of 59 US 9 , 783, 592 B2

Change in Body Weight Sahaky ABW(%) ??????? 1999- -

-300 2 4 6 8 10 12 14 16 18 20 22 Time (Days )

Vehicle GLP - 1 (AIB2 , E16 , Cex , K40 ) 40 ug /kg - day LLL- - 0 - - - GLP - 1 (AIB2 , E16 , Cex ,K40 YEstrogen (17 -ester ) 40 ug /kg -day TIL GLP - 1 (AIB ?, E16 ,Cex ,K40 YEstrogen ( 3 -ether ) 40 ug /kg -day GLP - 1 ( AIB2 ,E16 ,Cex , K40 ) 400ug /kg -day GLP - 1 (AIB2 , E16 ,Cex , K40 yEstrogen (17 -ester ) 400 ug/ kg -day + GLP - 1 (AIB2 , E16 ,Cex , K40 YEstrogen ( 3 -ether ) 400 ug /kg -day Fig . 4b U . S . Patent Oct. 10, 2017 Sheet 12 of 59 US 9 ,783 ,592 B2

Change in Fat Mass

• AFatMass(g) *

T ? ??? [J ] - 15 Day 21

O Vehicle LA GLP -1 ( AIB ?, E16 , Cex, K40) 40ug/ kg -day ZO GLP - 1 ( AIB2 , E16 , Cex , K40 Estrogen (17 -ester ) 40 ug /kg -day a GLP - 1 (AIB2 , E16, Cex, K40 )/ Estrogen ( 3 - ether) 40 ug/ kg - day 7 GLP - 1 (AIB , E16, Cex, K40 ) 400ug/ kg -day X GLP -1 ( AIB2, E16 , Cex , K40 )/ Estrogen ( 17 - ester) 400 ug /kg - day O GLP -1 (AIB2 , E16, Cex , K40 )/ Estrogen (3 - ether) 400 ug/ kg -day Fig . 4c atent Oct. 10, 2017 Sheet13 0f 59 US 9 , 783 ,592 B2

Change in Blood Glucose T ???? (p/bulogy ? ??????? ?????????? ???????? 77777777/771 ???? ? ??????????????? ???????? - 603 7 21 7 21 7 21 7 21 7 21 7 21 7 21 Time ( days)

O Vehicle Z4 GLP- 1 ( AIB2, E16, Cex, K40) 40ug/ kg -day 2 GLP- 1 ( AIB2, E16, Cex, K40 )/ Estrogen (17 -ester ) 40 ug/ kg - day GLP -1 ( AIB2, E16, Cex, K40 )/ Estrogen ( 3- ether) 40 ug/ kg -day kz GLP - 1 ( AIB2, E16, Cex, K40) 400ug / kg - day ?? GLP - 1 ( AiB2, E16, Cex, K40) / Estrogen (17 - ester ) 400 ug/ kg - day 24 GLP- 1 ( AIB2, E16 , Cex , K40 )/ Estrogen ( 3 - ether) 400ug /kg -day Fig . 4d atent Oct. 10 , 2017 Sheet 14 of 59 US 9 ,783 ,592 B2

Change in Body Weight 5

ABW(%)

- 20 - Time (Days ) * Vehicle QW Vehicle QD O GLP - 1 (AIB2 , E16 ,Cex ,K40 ) 40ug/ kg -day - 0 . - GLP - 1 (AIB2 , E16 ,Cex , K40 y/ Estrogen ( 3 - ether) 40 ug /kg - day - d - aa -GLP - 1 (A1 ,AIB ,A22 ,Cex ,K40 yEstrogen ( 3 - ether) 40 ug /kg -day GLP -1 (AIB2 , E16 ,C24 (PEG -40kDa ) , Cex ,K40 y/ Estrogen (3 - ether) 40 ug/ kg - day GLP - 1 (AIB2 , E16 , C24 (PEG -40kDa ) , Cex, K40 ) 40 ug/ kg - wk - -- - - GLP - 1 (AIB , E16 ,Cex , K40 ) 400ug /kg -day 4 GLP - 1 (AIB2 , E16 ,Cex ,K40 )/ Estrogen (3 - ether) 400 ug /kg -day d -aa -GLP - 1 (A1 , AIB2, A22 ,Cex , K40 )/ Estrogen ( 3 -ether ) 400 ug /kg -day a GLP - 1 (AIB2 , E16 ,C24 (PEG -40kDa ) ,Cex ,K40 \/ Estrogen ( 3 - ether )40 ug/ kg - wk Fig . 5a atent Oct. 10 , 2017 Sheet 15 of 59 US 9 ,783 ,592 B2

Change in Blood Glucose

Wh HZ ABG(mg/dL) HO WW ý udog ARARAAAAAAAA HAIIIII z Day 7

Vehicle QW O Vehicle QD 222 GLP - 1 (AIB2 , E16 ,Cex ,K40 ) 40 ug/ kg -day Z GLP - 1 ( AIB2 , E16 ,Cex , K40 YEstrogen ( 3 - ether ) 40 ug/ kg - day d - aa -GLP - 1 ( A1, AIB2, A22 ,Cex , K40 yEstrogen ( 3 -ether ) 40 ug /kg -day KZ GLP - 1 (AIB2 , E16 ,C24 (PEG - 40kDa) , Cex ,K40 YEstrogen ( 3 - ether) 40 ug /kg - day W GLP - 1 (AIB2 , E16 , C24 (PEG -40kDa ), Cex ,K49 ) 40 ug /kg -wk 2 GLP - 1 (AIB2 , E16 ,Cex ,K40 ) 400 ug /kg - day GLP - 1 ( AIB2, E16 ,Cex ,K40 YEstrogen ( 3 - ether ) 400 ug/ kg - day AW d -aa -GLP - 1 (A1 , AIB ? ,A22 , Cex , K40 YEstrogen ( 3 - ether) 400 ug /kg - day AV GLP - 1 ( AIB2, E16 , C24 (PEG - 40kDa ), Cex ,K40 YEstrogen ( 3 - ether ) 40 ug /kg -wk Fig . 5b atent Oct. 10 , 2017 Sheet 16 of 59 US 9 ,783 ,592 B2

Change in Body Weight

N o IKLUSH A BW(%) 10T A

áo

Time (Days )

Vehicle - - 0 - - GLP - 1 (AIB2 , E16 ,Cex , K40 ) 40ug/ kg -day d -aa -GLP - 1 (A1 , AIB2 , A22 , Cex , K40 ) 40 ug /kg -day GLP - 1 (AIB ? ,A22 , Cex , K40 ) 40 ug /kg - day GLP - 1 ( AIB2 , E16 ,Cex , K40 YEstrogen ( 3 -ether ) 40 ug /kg -day d - aa -GLP - 1 (A1 , AIB2 , A22 ,Cex ,K40 ) Estrogen ( 3 -ether ) 40 ug/ kg -day o GLP - 1 (AIB2 , 422 ,Cex , K40 )/ Estrogen ( 3 - ether ) 40ug/ kg -day - O GLP - 1 (AIB ? , E16 , Cex , K40 Estrogen ( 17 -ester ) 40 ug /kg - day - . . . - d -aa -GLP - 1 (A1 , AIB2, 422 , Cex, K40 )/ Estrogen ( 17 -ester ) 40 ug /kg -day ------GLP - 1 (AIB2 , A22 , Cex , K40 ) Estrogen ( 17 -ester ) 40 ug /kg -day Fig . 6a atent Oct. 10 , 2017 Sheet 17 of 59 US 9 ,783 ,592 B2

Fat Mass

one FatMass(g) ??????????h º 1111111 o ?????????? WMWWWW UNNNNNNNNM ZAIIIIIIZ Day HEILZILIZZA7

Vehicle Z GLP - 1 ( AIB ?, E16 ,Cex ,K40 ) 40 ug /kg -day Z d -aa -GLP - 1 (A1 , AIB2 , A22 ,Cex ,K40 ) 40 ug /kg -day W GLP - 1 (AIB2 , A22 , Cex , K40 ) 40 ug /kg -day B2 GLP - 1 (AIB2 , E16 ,Cex , K40 Estrogen ( 3 - ether ) 40 ug /kg -day W d - aa -GLP - 1 (A1 , AIB ? ,A22 , Cex , K40 y/ Estrogen ( 3 - ether) 40 ug/ kg -day Z GLP - 1 ( AIB ? , A22, Cex ,K40 )/ Estrogen ( 3 - ether ) 40 ug /kg - day GLP - 1 (AIB2 , E16 ,Cex ,K40 YEstrogen ( 17 - ester ) 40 ug /kg -day W d - aa -GLP - 1 (A ?, AIB2 , A22 ,Cex , K40 )/ Estrogen ( 17 -ester )40 ug /kg - day N GLP - 1 (AIB2 , A22 ,Cex ,K40 )/ Estrogen (17 - ester) 40 ug/ kg -day Fig . 6b atent Oct. 10 , 2017 Sheet 18 of 59 US 9 ,783 ,592 B2

Change in Blood Glucose ABG(mg/dL) - TV |? MW Z2 [? ??

Day 7

Vehicle Z GLP - 1 (AIB2 , E16 ,Cex , K40 ) 40ug /kg -day Z d -aa -GLP - 1 (A ', AIB2 , A22 ,Cex , K40 ) 40 ug/ kg -day W GLP - 1 (AIB2 , A22 ,Cex , K40) 40 ug /kg -day 12 GLP - 1 (AIB ? , E16 ,Cex ,K40 YEstrogen ( 3 - ether ) 40 ug /kg -day V d - aa -GLP - 1 (A1 , AIB ? ,A22 , Cex , K40 )/ Estrogen (3 - ether ) 40 ug /kg -day Z GLP - 1 (AIB ? ,A22 , Cex , K40 )/ Estrogen ( 3 -ether ) 40 ug /kg -day GLP - 1 (AIB2 , E16 ,Cex ,K40 YEstrogen ( 17 - ester ) 40 ug /kg -day WW d -aa -GLP - 1 ( A ’, AIB2 , 422 ,Cex , K40 YEstrogen ( 17 -ester ) 40 ug/ kg -day N GLP - 1 ( AIB2, A22 ,Cex , K40 yEstrogen (17 -ester ) 40 ug /kg -day Fig . 6c atent Oct. 10 , 2017 Sheet 19 of 59 US 9 ,783 ,592 B2

Change in Body Weight

Bodyweight(%) · - - - - •

2 6 8 Time (Days ) + Vehicle O GLP - 1 (AIB2 , E16 ,Cex , K40 ) 400 ug /kg -day - - - GLP - 1 (AIB2 , E16 ,Cex , K40 yEstrogen ( 3 - ether) 400 ug /kg -day * GLP - 1 (AIB2 , E16 , Cex, K40 yEstrogen ( 17 -ester ) 400 ug/ kg -day 7 d - aa GLP - 1 (A1 , AIB2 , A22, Cex , K40 ) 400 ug /kg - day + d -aa GLP - 1 ( A ', AIB , A22 , Cex , K40 yEstrogen ( 3 - ether ) 400 ug /kg -day + d -aa GLP - 1 ( A !, AIB2, A22 , Cex , K40 yEstrogen (17 -ester ) 400 ug/ kg -day + GLP - 1 (AIB2 , A22 ,Cex ,K40 ) 400 ug/ kg -day + GLP - 1 (AIB2 , A22 ,Cex , K40 YEstrogen ( 3 -ether ) 400 ug/ kg - day + GLP - 1 (AIB , A22 ,Cex ,K40 yEstrogen ( 17 -ester ) 400 ug /kg - day Fig . 7a atent Oct. 10 , 2017 Sheet 20 of 59 US 9 ,783 ,592 B2

Change in Blood Glucose

ABG(mg/dL) opogå TYYNN

Day 7

Vehicle Z GLP - 1 (AIB2 , E16 ,Cex ,K40 ) 400 ug /kg -day Z GLP - 1 (AIB2 , E16 ,Cex ,K40 YEstrogen (3 -ether ) 400 ug /kg - day E GLP - 1 (AIB2 , E16 ,Cex , K40 y/ Estrogen ( 17 -ester ) 400 ug /kg -day Z d -aa GLP - 1 ( A1, AIB2 ,A22 ,Cex ,K40 ) 400 ug/ kg -day N d -aa GLP - 1 (A ', AIB2, A22 ,Cex , K40 yEstrogen ( 3 -ether ) 400 ug /kg -day z d -aa GLP - 1 ( A ”, AIB2, A22, Cex , K40 /Estrogen (17 - ester ) 400 ug/ kg - day N GLP - 1 (AIB2 , A22 , Cex , K40 ) 400 ug /kg -day W GLP - 1 (AIB2 , A22 , Cex , K40 ) Estrogen ( 3 - ether) 400 ug /kg -day N GLP - 1 (AIB ? , A22 , Cex , K40 YEstrogen ( 17 -ester ) 400 ug /kg -day Fig . 7b atent Oct. 10 , 2017 Sheet 21 of 59 US 9 ,783 ,592 B2

Change in Body Weight

? ABW(%) IN2 -

_ FONK - - - - - _

? Time (Days )

- Vehicle O GLP - 1 (AIB2 , E16 ,Cex , K40 ) 120ug /kg - day - GLP - 1 ( AIB ? ,E16 , Cex , K40) 400 ug /kg -day - A . GLP - 1 (AIB ? , E16 ,Cex ,K40 \ Estrogen ( 3 - ester ) 120 ug /kg - day - * . GLP - 1 (AIB2 , E16 ,Cex , K40 yEstrogen ( 3 - ester ) 400 ug /kg -day + d - aa GLP - 1 ( A1, AIB2, A22, Cex , K40 )400ug /kg -day + d -aa GLP - 1 (A1 , AIB ?, A22 ,Cex ,K40 ) 1 ,200 ug /kg -day - : d -aa GLP - 1 ( A ' ,AIB2 , A22 ,Cex ,K40 yEstrogen ( 3 - ester ) 400 ug /kg -day * d - aa GLP - 1 (A1 , AIB2 , A22, Cex ,K40 Estrogen ( 3 - ester ) 1 , 200 ug /kg - day Fig . 8a U .S . Patent Oct. 10, 2017 Sheet 22 of 59 US 9 , 783, 592 B2

Change in Blood Glucose Levels

ABG(mg/dL) No A HA HA HW

Day 7

Vehicle W GLP - 1 (AIB2 , E16 ,Cex , K40 ) 120 ug /kg -day Z GLP - 1 (AIB2 , E16 , Cex ,K40 ) 400ug / kg -day K GLP - 1 ( AIB2, E16 ,Cex , K40 YEstrogen ( 3 -ester ) 120 ug/ kg - day WZ GLP - 1 (AIB2 , E16 , Cex , K40 YEstrogen ( 3 -ester ) 400 ug /kg -day N d - aa GLP - 1 (A1 , AIB2, A22 ,Cex ,K40 ) 400 ug /kg -day Z d - aa GLP - 1 ( A1, AIB2, A22 ,Cex , K40 ) 1 , 200 ug /kg - day d -aa GLP - 1 (A1 , AIB2 , A22 , Cex , K40 YEstrogen ( 3 - ester) 400 ug/ kg -day W d -aa GLP - 1 (A1 , AIB2, A22 ,Cex , K40 YEstrogen ( 3 - ester) 1 , 200 ug /kg - day

Fig . 8b U .S . Patent Oct. 10, 2017 Sheet 23 of 59 US 9 , 783, 592 B2

Fat Mass FatMass(g) WWW ??????? WWWWWW III VIII 7777772 Day 7

Vehicle WZ GLP - 1 ( AIB ? ,E16 ,Cex , K40 ) 120ug /kg -day Z GLP - 1 ( AIB2 ,E16 ,Cex , K40 ) 400 ug /kg -day W GLP - 1 ( AIB2, E16 ,Cex , K40 \/ Estrogen ( 3 -ester ) 120 ug /kg -day WZ GLP - 1 (AIB2 , E16 , Cex , K40 YEstrogen ( 3 - ester) 400 ug /kg -day N d -aa GLP - 1 (A1 , AIB2 , A22, Cex , K40 ) 400ug /kg -day z d - aa GLP - 1 ( A ? ,AIB2 , A22 ,Cex , K40) 1 , 200 ug/ kg - day daa GLP - 1 (A1 , AIB ?, A22 ,Cex ,K40 y/ Estrogen ( 3 -ester ) 400 ug /kg -day W d - aa GLP - 1 (A1 , AIB2, A22 ,Cex ,K40 )/ Estrogen ( 3 - ester ) 1 , 200 ug/ kg - day

Fig . 8c atent Oct. 10 , 2017 Sheet 24 of 59 US 9 , 783 ,592 B2

Change in Body Weight

ABW(%) ú . Time (Days ) s

+ Vehicle O GLP - 1 (AIB2 , E16 ,Cex , K40 )/ Estrogen ( 3 -ether ) 40 ug /kg -day + d -aa GLP - 1 (A1 , AIB2 , A22, Cex , K40 ) 4000 ug /kg -day + d -aa GLP - 1 ( A ', AIB ? , A22 ,Cex , K40 )/ Estrogen ( 3 -ester ) 400 ug /kg -day + d -aa GLP - 1 (A1 , AIB2, A22 ,Cex , K40 ) /Estrogen ( 3 -ester ) 1200 ug /kg -day d -aa GLP - 1 ( A ”, AIB2, A22 ,Cex , K40 \/ Estrogen ( 3 -ester ) 4000 ug/ kg -day + GLP - 1 (AIB2 , E16 , C24 (PEG -40kDa ) ,Cex , K40 ) 40 ug /kg -day - - - GLP - 1 (AIB2 , E16 , C24( PEG -40kDa ) , Cex , K40 ) /Estrogen ( 3 - ether ) 40 ug/ kg -day GLP -1 (AIB2 , E16 , C24 (PEG - 40kDa ), Cex , K40 )/ Cholesterol ( 3 - ether) 40 ug /kg -day Fig . 9a atent Oct. 10 , 2017 Sheet 25 of 59 US 9 ,783 ,592 B2

Fat Mass

FatMass(g) VIININ / VINVIIN ? VIIN VIIN / VIINVIII 77777777777 Day 7

Vehicle Z GLP - 1 (AIB2 , E16 ,Cex ,K40 )/ Estrogen (3 -ether ) 40 ug /kg -day Z d - aa GLP - 1 (A1 , AIB2, A22, Cex , K40 ) 4000 ug /kg - day w d -aa GLP - 1 ( A , AIB ? , A22, Cex , K40 )/ Estrogen ( 3 -ester ) 400 ug/ kg - day WZ d - aa GLP - 1 ( A1, AIB ? , A22 ,Cex , K40 ) /Estrogen ( 3 -ester ) 1200 ug /kg -day N d -aa GLP - 1 (A ', AIB ? , A22, Cex , K40 )/ Estrogen ( 3 -ester ) 4000 ug/ kg - day Z GLP - 1 ( AIB2, E16 , C24 (PEG - 40kDa) , Cex, K40 ) 40 ug/ kg - day N GLP - 1 (AIBP 2 , E16 , C24 (PEG -40kDa ) , Cex , K40 /Estrogen (3 - ether) 40 ug/ kg - day WW GLP - 1 (AIB ?, E16 , C24 (PEG -40kDa ), Cex, K40 y/ Cholesterol ( 3 -ether ) 40 ug/ kg -day Fig . 9b U. S . Patenatent Oct. 10 , 2017 sketSheet 26286 of015 59 US 9 ,783 ,592 B2

Change in Blood Glucose Levels

98V)bu/1p(

Day 7 - Day o Vehicle Z GLP - 1 (AIB2 , E16 ,Cex ,K40 )/ Estrogen ( 3 - ether) 40 ug /kg -day Z daa GLP - 1 (A1 , AIB2, A22 ,Cex , K40 ) 4000 ug/ kg - day d -aa GLP - 1 ( A1, AIB2, A22, Cex , K40 )/ Estrogen ( 3 -ester ) 400 ug /kg -day 2 d - aa GLP - 1 ( A ” , AIB2, A22 , Cex , K40 ) /Estrogen ( 3 -ester ) 1200 ug /kg -day N d -aa GLP -1 ( A ', AIB2, A22 ,Cex ,K40 )/ Estrogen (3 -ester ) 4000 ug/ kg -day Z GLP - 1 (AIB2 , E16 , C24 (PEG -40kDa ) , Cex, K40 ) 40 ug /kg -day GLP - 1 (AIB2 , E16 , C24 (PEG -40kDa ) , Cex ,K40 )/ Estrogen ( 3 -ether ) 40 ug /kg - day AW GLP - 1 (AIB2 , E16 , C24( PEG -40kDa ), Cex ,K40 )/ Cholesterol (3 -ether ) 40 ug /kg - day Fig . 9c U . S . Patent Oct. 10 , 2017 Sheet 27 of 59 US 9 , 783, 592 B2

Change in Body Weight

_ Ama 10

ABW(%) enMa?

Time ( Days )

+ Vehicle o d -aa GLP - 1 ( A ? , AIB2, A22 ,Cex , K40 yEstrogen ( 3 -ester ) 1 , 200 ug /kg - day a GLP - 1 ( A ' , AIB2 , A22 ,Cex , K40 Estrogen ( 3 -ether ) 1 , 200 ug/ kg -day + d - aa GLP - 1 (AIB ? , E16 ,Cex , K40 ) 4 , 000 ug/ kg -day od- aa GLP - 1 (A " , AIB ? , A22 ,Cex , K40 )/ Estrogen ( 3 - ester) 4 , 000 ug /kg -day + d - aa GLP - 1 (A ' , AIB2, A22 ,Cex , K40 Estrogen ( 3 - ether ) 4 ,000 ug/ kg -day Fig . 10a atent Oct. 10 , 2017 Sheet 28 of 59 US 9 ,783 ,592 B2

Change in Blood Glucose Levels

lii ABG(mg/dL)

ITTDay 7 -Day 0 Vehicle W d -aa GLP - 1 ( A , AIB2, A22 , Cex, K40 )/ Estrogen (3 - ester) 1 ,200 ug /kg -day z d -aa GLP - 1 ( A ', AIB2, A22, Cex , K40 YEstrogen ( 3 - ether) 1 ,200 ug /kg -day W d -aa GLP - 1 (AIB2 , E16 ,Cex ,K40 ) 4 , 000 ug /kg -day W d -aa GLP - 1 ( A " , AIB2, A22 ,Cex , K40 YEstrogen ( 3 - ester) 4 ,000 ug /kg -day V d - aa GLP - 1 (AT , AIB2, A22, Cex ,K40 ) Estrogen ( 3 - ether) 4 ,000 ug /kg -day Fig . 10b atent Oct. 10 , 2017 Sheet 29 of 59 US 9 ,783 ,592 B2

GLP - 1 Receptor Activation 4000 GLP - 1 3 ' Ether

3500 - A 17' Ester . ¥ 3 ' Activated Ester 17 ' OAC HD DRERIROCOOOOOOO 17 ' Hydrazone ATUN 3000 - 17 ' Carbamate Disulfide **** * * * +7137 raminif 2500 ti{

H Te

. titu{{} } } } }{ tutitut????????{ .

Luminescence . 1000 .

. 500 Stories Sinulle1150RHamilton # su ques to 1E - 4 1E - 3 0 .01 0 . 1 Concentration , nM Fig . 11a atent Oct. 10 , 2017 Sheet 30 of 59 US 9 ,783 ,592 B2

3-Ester17OAC 17-Hydrazone 17-CarbamateDisulfide Estradiol 3-Ether 17-Ester A # À 1000 ee . foweb dan ???????????? 100

10 fundamay XE

------in LinkerStability Concentration,nM 0.1 Fig.11b H ???????????????? A 0.01 *

1E-3 framebord

1E-4

ebm boneconw bersamaundmannalaunenwandamannaalamandrama where * 8 ) Luminescence atent Oct. 10 , 2017 Sheet 31 of 59 US 9 ,783 ,592 B2

Change in Body Weight Oproteoog TO ABW(%) 5578KV -3 - 5 - 30 2 4 6 8 10 12 14 Time ( days )

Vehicle O d -amino acid GLP - 1 - ~ . d -amino acid GLP - 1 /Estrogen ester 400 ug /kg -day - - d - amino acid GLP - 1 / Estrogen ester 4 ,000 ug/ kg -day - O d - amino acid GLP - 1 /Estrogen ether 400 ug /kg -day + d - amino acid GLP - 1 /Estrogen ether 4 ,000 ug /kg -day Fig . 12a U . S . Patent Oct. 10 , 2017 Sheet 32 of 59 US 9 , 783, 592 B2

Food Intake

FI(g) Ñ tha

TTTT 0 2 4 6 8 10 12 14 Time ( days )

– Vehicle Od- - amino acid GLP - 1 - ~ - d - amino acid GLP - 1 /Estrogen ester 400 ug/ kg -day 1 - d - amino acid GLP - 1 /Estrogen ester 4 ,000 ug/ kg -day - O - d - amino acid GLP - 1 /Estrogen ether 400 ug /kg -day A d -amino acid GLP - 1 /Estrogen ether 4 ,000 ug/ kg -day Fig . 12b U . S . Patent Oct. 10 , 2017 Sheet 33 of 59 US 9 , 783, 592 B2

Change in Blood Glucose

ABG(mg/dL) HAI O

Day 14

Z Vehicle 2 d -amino acid GLP - 1 z d -amino acid GLP - 1 /Estrogen ester 400 ug /kg -day w d -amino acid GLP - 1/ Estrogen ester 4 ,000 ug /kg -day WZ d -amino acid GLP - 1 /Estrogen ether 400 ug /kg -day N d -amino acid GLP - 1 /Estrogen ether 4 ,000 ug /kg -day Fig . 12c atent Oct. 10 , 2017 Sheet 34 of 59 US 9 ,783 ,592 B2

Liver weight

Weight(g) IIIA Allu IN III LIIIII III IIIII / Day 14

Z Vehicle W d -amino acid GLP - 1 Z d - amino acid GLP - 1 /Estrogen ester 400 ug /kg -day Be d - amino acid GLP - 1 /Estrogen ester 4 ,000ug /kg -day WZ d -amino acid GLP - 1 /Estrogen ether 400ug /kg -day SV d -amino acid GLP - 1 /Estrogen ether 4 ,000ug /kg -day Fig . 12d U .S . Patenatent Oct. 10 , 2017 Sheet 35 of 59 US 9 ,783 ,592 B2

Uterus weight 0. 87

Weight(g) Ho VIIIIII VIII

IIIIII /

0 . 0 Day 14

Z Vehicle W d -amino acid GLP - 1 Z d -amino acid GLP - 1 /Estrogen ester 400 ug /kg - day By d -amino acid GLP - 1 /Estrogen ester 4 ,000 ug /kg -day WZ d -amino acid GLP - 1 /Estrogen ether 400 ug /kg - day V d -amino acid GLP - 1 /Estrogen ether 4 ,000 ug /kg - day Fig . 12e atent Oct. 10 , 2017 Sheet 36 of 59 US 9 ,783 ,592 B2

Change in Body Weight

FE I 8 (%)MAY ------De het Time (Days ) Vehicle GLP - 1 Agonist . .. . 1 ...... d -amino acid GLP - 1 /Estrogen ester d -amino acid GLP - 1 /Estrogen ether - - - GLP -1 / Estrogen ether Fig . 13a U .S . Patentatent Oct. 10 , 2017 Sheet 37 of 59 US 9 , 783 ,592 B2

1000d-aminoacidGLP1/Estrogenester d-aminoacidGLP1/Estrogenether GLP-1Agonist OGLP-1Estrogen/ether OVehicle IIIIII TTTTTTMUUUU TII IIIIIIITTTTTTT TTTTTTT IIIIIII TTTTTTTLIITTI TI

ChangeinFatMass Day7-0 Fig.13B ? and0.47 -0.4 -0.6 -0.87 ) g ( MassFat A atent Oct. 10 , 2017 Sheet 38 of 59 US 9 ,783 ,592 B2

Cumulative Food Intake

-- - - - FI(g) - -

2 4 SecaTime (Days ) Vehicle GLP - 1 Agonist ...... d - amino acid GLP - 1 /Estrogen ester - - - A- - - d -amino acid GLP -1 /Estrogen ether TU- 0 - GLP - 1 /Estrogen ether FIG . 13C atent Oct. 10 , 2017 Sheet 39 of 59 US 9 ,783 ,592 B2

esterEstrogenacidGLP-1/amino000d d-aminoacidGLP1/Estrogenether GLP-1Agonist GLP-1/Estrogenether VehicleO

E IIIIIIITTTTTTT TTTTTTT TTTTTT! TTTTTTT IIIIIII TTTTTT! IIIIIII IIIIIII IIIIIII IIIIIII TTTTTTT IIIIIII IIIIIII TTTTTTT

ChangeinBloodGlucoseLevels Day7-0 Fig.13D H ? ? ? ? ? ? ) dL/ mg ( ABG U .S . Patentatent Oct. 10 , 2017 Sheet 40 of 59 US 9 ,783 ,592 B2

esterEstrogenGLP-1/acidamino10d 8d-aminoacidGLP1/Estrogenether GLP-1Agonist it!GLP-1/Estrogenether OVehicle

??????IIIIIII TTTTTTT TTTTTT TTTTTTTIIIIIII IIIIII III| TTTTTTT

UterusWeight Day7 FIG.13E

0.15 0.10? ) g ( Weight Uterus U .S . Patentatent Oct. 10 , 2017 Sheet 41 of 59 US 9 ,783 ,592 B2

GLP-1Agonist/E2cathepsinlabile OGLP-1Agonist/Estrone3ester -GLP1Agonist/E217carbamatedisulfide +GLP-1Agonist/E217hydrazone

*Vehicle

H Ri Mat BodyWeight(%) Time(Days) FIG.14A

? 1 10 ) % ( BW 10- U .S . Patentatent Oct. 10 , 2017 Sheet 42 of 59 US 9 ,783 ,592 B2

GLP-1Agonist/Estrone3ester MOI1GLP-1Agonist/E217carbamatedisulfide3GLP-1Agonist/E217hydrazone NGLP-1Agonist/E2cathepsin-labile

OVehicle

Time(Day7) FatMass FIG.14B

A Ì ) 9( Mass Fat U .S . Patentatent Oct. 10 , 2017 Sheet 43 of 59 US 9 ,783 ,592 B2

GLP-1Agonist/Estrone3-ester DIGLP-1Agonist/E217carbamatedisulfideEGLP-1Agonist/E217hydrazone NGLP-1Agonist/E2cathepsinlabile

DVehicle

BloodGlucose Time(Days) FIG.14C

) dL/ mg ( BG U .S . Patentatent Oct. 10 , 2017 Sheet 44 of 59 US 9 ,783 ,592 B2

esterEstrone-3Agonist/GLP1- MINNGLP-1Agonist/E217carbamatedisulfide EGLP-Agonist1/E217hydrazone NGLP-1Agonist/E2cathepsinlabile

OVehicle

UterusWeights Time(Day7) FIG.14D

0.5 0.1 0. ) g ( Uterus atent Oct. 10 , 2017 Sheet 45 of 59 US 9 ,783 ,592 B2

Dose 40ug/kgday 400ug/kgday 40ug/kgday 40ug/kgdayo 40ug/kgday 400ug/kgday

GLP-1Agonist/Estrogen(3ether). GLP-1Agonist/Estrogen(3ether) GLP-1Agonist Compound -AgonistGLP-1 GLP-1Agonist/Estrogen(3ester) GLP-1Agonist/Estrogen(3ester)– VehicleVehicle

???? 4

I RMA?? Time(Days) 47 tet 02468101214 FIG.15A

8 8 8 8 -30TTTTTTTT ) % ( Weight Body A U . S . Patent Oct. 10 , 2017 Sheet 46 of 59 US 9 ,783 ,592 B2

Dose 40ug/kgday 400ug/kgday ug/kgday40 ug/kgdayo40 ug/kgday40 ug/kgday400

.Estrogen(3-ether)Agonist/|GLP1 GLP-1Agonist/Estrogen(3ether) -AgonistGLP1 Estrogen(3-ester)Agonist/GLP1 Estrogen(3-ester)Agonist/GLP1 Compound Agonist-GLP1 Vehicle 15BFIG. KK ? K ? 14 Hy 12 ...... of- 10 n Time(Days) 8

6 tiimimisto y 4 SAATIITSHIRTTTTTT2 etc0 ?eo ñ ? ò oor ) g ( Intake Food A atent Oct. 10 , 2017 Sheet 47 of 59 US 9 ,783 ,592 B2

GLP-1/Estrogen(3ester)(Estrogen-Labile)

FIG.16A

, GLP-1/Estrogen(3ether)(Estrogen-Stable)

,. SZAFY ? U. S . Patentatent Oct. 10 , 2017 Sheet 48 of 59 US 9 , 783 ,592 B2

ww mmmm Ohr WWWW 3hrs 6hrs wwwwwwwwwwww new XXXL WYDA tas 37°C.0pH5 WWWWINDOWS WWWhere WoossenSUOSITUTLLLL wwwwwwwwwwwwco.ca wmmmmmmmmmmmmmyyyyyyymimmmm COOK si wwwwwww w

ALLALLWARN A myMyN 44444444444444 * WAUWWW GLP-1/Estrogen(17hydrazone)AcidLabile PUZWZWMUWX2127ZW222WWZWX4222WUZUMUWM FIG.16B Ohr 12hrs 24hrs 24112012444444244864424W4*&22144

VN W SVALYUSY-Yuur ??????????????????? WWW. pH7.437°C

4 WWWWWWWWWWWWWWWWWWWWWWWWWWWWWWWWWWWWWWWWWWWWWWW 24*4 atent Oct. 10 , 2017 Sheet 49 of 59 US 9 ,783 ,592 B2

???????????????????????????????????????????????????????????? ??? Low Ohr 24hrs 48hrs ee????????????????????????????????www.wervrown2-r1

a HinderedThiolReduction-Labile * * W * Coco 10 WXNm VULVVVVVVV wieci eriwwrenm

tuuLLLLLLLLLLLLLLLLLLLLLL LUUUUUUUUUUUUUUUUUUUUUUUUUUUUUUUUU FIG.16C GLP-1/Estrogen(17carbamatedisulfide) ??????????????????????????????????????????????? ????????????????????????????????????????????????????????????????? Ohr 24hrs 48hrs

INTRO PPPPPPPPPPPPPPRZYWZZZ .

w tay ASUNA S - se TIT UnhinderedThiolReduction-Labile .. .WWW w - ' emi rd wwwwwwwwwwwVINA OYNUFISALIPPPPPPPPP7290VXPWPPPP202777PP27772W WWUNAWZumP w *

???????????????????????????????????????????? U . S . Patent Oct. 10 , 2017 Sheet 50 of 59 US 9 , 783 , 592 B2

Ohr 3hrs 6hrs

37°CGlutathionemM15

Labile)Reduction-ThiolHindered( Intracellular GLP1/Estrogen-(17carbamatedisulfide) 16D.FIG Ohr 12hrs 24hrs

www am 37°CGlutathioneUM15 Extracellular U . S . Patent Oct . 10, 2017 Sheet 51 of 59 US 9 ,783 ,592 B2

GLP - 1 /Estrogen (cathepsin ) (Enzyme - Labile )

0 hrs

12 hrs

24 hrs

Human Plasma 37°C FIG . 16E U .S . Patenatent Oct. 10 , 2017 Sheet 52 of 59 US 9 ,783 ,592 B2

ether)Estrogen(3-Agonist/Glucgon GIPAgonist/Estrogen(3-ether) GLP-1Agonist/Estrogen(3ether) GlucagonAgonist Vehicle GIPAgonist GLP-1Agonist

D O O A O •

. o

. g ? ? ) % ( ABW ? 8 U .S . Patentatent Oct. 10 , 2017 Sheet 53 of 59 US 9 ,783 ,592 B2

GlucgonAgonist/Estrogen(3-ether) GIPAgonist/Estrogen(3-ether) GLP-1Agonist/Estrogen(3ether) GlucagonAgonist Vehicle GIPAgonist GLP-1Agonist TT

II| O 0000010 D IIIIIII

IIIIIIIIIIIIIIIII IIIIIIIIIIIIIIIIIIIII 11 IIIIIIIIIIIIIIIIII TUOTTOTITOLUL ?? ?????????????????

Time(Day7) TotalFoodintake(PerMouse) FIG.17B

? ? ? ? 1 2 ) g( intake Food U .S . Patentatent Oct. 10 , 2017 Sheet 54 of 59 US 9 ,783 , 592 B2

GlucgonAgonist/Estrogen(3-ether) GIPAgonist/Estrogen(3-ether) GLP-1AgonistEstrogen/(3ether) GlucagonAgonist Vehicle GIPAgonist GLP-1Agonist

|0 III - B IIIIIII ITTTTTT

LITTTTTTTTTTTTTTTTTTTTT TOTUOTTOTITOLULUI IIIIIIIIIIIIIIIIIIIII TTTTTTTTTTTTTTTTTTTTTTTT ITULLIITTO TTTIIIIIIIIIIIIIIIIIIIIIII Lumil UTITI ILTILL TIITTI ULTIMITI TUTTI = = = HALI| | || | | | | | | | | | | | | | | | | | || WWIINIZ Time(Day0,7) BloodGlucose A TITI FIG.17C HU

BGLP-1Agonist/Estrogen(3ether)ERBKO EGLP-1Agonist/Estrogen(3ether)WTERKO NGLP-1Agonist/Estrogen(3ether)ERAKO

OVehicleERBKO MINIGLP-1AgonistERBKO NVehicleWTERKO HGLP-1AgonistWTERKO 6VehicleERAKO EGLP-1AgonistEraKo

HIIT IIIIIII 111111111 14+ELHH+EH TTDI

LJILI. HELFitten OLIIT

Time(Day14) TotalChangeinBodyWeight(%) FIG.18A

AGLP-1Agonist/Estrogen(3ether)ERBKO •GLP-1Agonist/Estrogen(3ether)WTERKO VGLP-1Agonist/Estrogen(3ether)ERAKO

OGLP-1AgonistERBKO VehicleERaKO IGLP-1AgonistERAKO OVehicleERBKO oVehicleWTERKO VGLP-1AgonistWTERKO

CumulativeFoodIntake Time(Days) FIG.18B

40- ? ô )9 ( Intake Food U .S . Patentatent Oct. 10 , 2017 Sheet 57 of 59 US 9 ,783 ,592 B2

3GLP-1Agonist/Estrogen(ether)ERBKO GLP-1Agonist|Estrogen(3ether)WTERKO WGLP-1Agonist/Estrogen(3ether)ERAKO GLP-1AgonistWTERKO OVehicleERBKO MIMIGLP-1AgonistERBKO SVehicleWTERKO 3VehicleERAKO EGLP-1AgonistERAKO

TTT E

11 E 111 JL+21HH+FT ITI ETCH

FETESTHE???f?? 01711 LLLLLL IIIIIIII LITT + + + TTTTTT DIIIIIIII

Time(Day14-0) GlucoseBloodChangein MINUMW JUTITI IIIIIII MITTITT MITIMITIIIIIII MUULUT FIG.18C

HE FH § $ $ ) dL/ ml ( BG atent Oct. 10 , 2017 Sheet 58 of 59 US 9 ,783 , 592 B2

TOU

40 50nmol/kgGLP-1AgonistEstrogen(3ester) 243.- -A50nmol/kgGLP1AgonistEstrogen(3ether)

-+50nmol/kgGLP1 32 . vehicle

24 Hours FIG.19

- Home o 8 8 8 8 8 8 o ) dL/ mg ( Glucose atent Oct. 10 , 2017 Sheet 59 of 59 US 9 ,783 ,592 B2

Fig . 20A Linking Group ( L )

Fig . 20B A Hydrophobic Linking Group B

Fig . 200

Hydrophilic Linking Group US 9 , 783 , 592 B2

GLUCAGON SUPERFAMILY PEPTIDES tinal growth , as well as the regulation of food intake . EXHIBITING NUCLEAR HORMONE Glucagon is a 29- amino acid peptide that corresponds to RECEPTOR ACTIVITY amino acids 33 through 61 of pre -proglucagon , while GLP - 1 is produced as a 37 - amino acid peptide that corresponds to CROSS REFERENCE TO RELATED amino acids 72 through 108 of pre - proglucagon . GLP - 1 ( 7 36 ) amide or GLP- 1 ( 7 - 37 ) acid are biologically potent forms APPLICATIONS of GLP -1 , that demonstrate essentially equivalent activity at This application is a continuation of U . S . application Ser. the GLP - 1 receptor. No . 13 /697 ,018 , filed Nov . 9 , 2012 , which is a U . S . national Glucagon is used in the acute treatment of severe hypo counterpart application of international application Ser. No. 10 glycemia . Oxyntomodulin has been reported to have phar PCT/ US2011 / 035913 filed May 10 , 2011, which claims macological ability to suppress appetite and lower body priority to U . S . Provisional Patent Application Nos. 61/ 334 . weight. GLP - 1 and GLP - 1 receptor agonists are used as 435 . filed on May 13 , 2010 and 61/ 432. 077 , filed on Jan . 12 . treatment for Type II diabetes . Exendin - 4 is a peptide present 2011. The entire disclosures of PCT /US2011 /035913 , U . S . in the saliva of the Gila monster that resembles GLP - 1 in Ser. No. 61 / 334 , 435 , and U . S . Ser. No. 61 / 432 , 077 are 15 structure , and like glucagon and GLP - 1 , increases insulin hereby incorporated by reference in their entirety . release . Gastric inhibitory polypeptide (GIP ) is also known as a INCORPORATION BY REFERENCE OF glucose -dependent insulinotropic peptide , and is a member MATERIAL SUBMITTED ELECTRONICALLY of the secretin family of hormones. GIP is derived from a 20 153 -amino acid proprotein encoded by the GIP gene and Incorporated by reference in its entirety is a computer circulates as a biologically active 42- amino acid peptide . readable amino acid sequence listing submitted concurrently The GIP gene is expressed in the small intestine as well as herewith and identified as follows: One 889 ,723 byte ASCII the salivary glands and is a weak inhibitor of gastric acid ( Text ) file named “ 238513SEQLIST . txt" created on Jul. 23 , secretion . In addition to its inhibitory effects in the stomach , 2015 . 25 in the presence of glucose, GIP enhances insulin release by pancreatic beta islet cells when administered in physiologi BACKGROUND OF THE INVENTION cal doses . GIP is believed to function as an enteric factor that stimulates the release of pancreatic insulin and that may play Field of the Disclosure a physiological role in maintaining glucose homeostasis . This invention provides glucagon superfamily peptides 30 Osteocalcin is a noncollagenous protein found in bone conjugated to nuclear hormone receptor ligands that are and dentin . It is secreted by osteoblasts and thought to play capable of acting at nuclear hormone receptors . a role in mineralization and calcium ion homeostasis . Osteo Brief Description of Related Technology calcin has also been reported to function as a hormone in the Nuclear hormone receptor proteins form a class of ligand body, causing beta cells in the pancreas to release more activated proteins that, when bound to specific sequences of 35 insulin , and at the same time directing fat cells to release the DNA , serve as on - off switches for transcription within the hormone adiponectin , which increases sensitivity to insulin . cell nucleus. These switches control the development and differentiation of skin , bone and behavioral centers in the SUMMARY OF THE INVENTION brain , as well as the continual regulation of reproductive tissues. 40 Provided herein are glucagon superfamily peptides con Nuclear hormone receptor ligands such as steroids , ste - jugated to nuclear hormone receptor ligands (“ NHR rols , retinoids, thyroid hormones , and vitamin D function to ligands” ) . These conjugates with plural activities are useful activate nuclear hormone receptors . The interaction of the for the treatment of a variety of diseases. hormone and receptor triggers a conformational change in The glucagon superfamily peptide conjugates of the the receptor, which results in the up -regulation of gene 45 invention can be represented by the following formula : expression . The level of cellular signal transduction acti vated by the interaction of a ligand and a nuclear hormone Q- L - Y receptor is determined by the number of ligands and recep - wherein Q is a glucagon superfamily peptide, Y is a NHR tors available for binding , and by the binding affinity ligand , and L is a linking group or a bond . between the ligand and the receptor. Many ligands and 50 The glucagon superfamily peptide ( Q ) in some embodi corresponding analogs that bind to nuclear hormone recep - ments can be a glucagon related peptide that exhibits agonist tors are used as medication to treat , for example , Parkinson ' s activity at the glucagon receptor, agonist activity at the disease (NURR1 ), sleep disorders (RZRB ), arthritis and GLP - 1 receptor, agonist activity at the GIP receptor, co cerebellar ataxia (RORa ) , central nervous system disorders agonist activity at the glucagon and GLP - 1 receptors, co (NOR - 1, Rev - ErbAB , Tlx , NGFI- BB , HZF - 2a , COUP - TFa , 55 agonist activity at the glucagon and GIP receptors , co COUP - TFB , COUR - TFY , NUR77 ) , hypercholesterolemia agonist activity at the GLP - 1 and GIP receptors, or tri (LXRa , COR ), obesity (Rev - ErbAa ) , diabetes (HNF4a ) , agonist activity at the glucagon , GIP , and GLP - 1 receptors . immune disorders ( TOR ) , metabolic disorders (MB67a , In some embodiments , the glucagon related peptide exhibits SHP, FXR , SF - 1 , LXRB ) , and infertility and contraception antagonist activity at the glucagon , GLP - 1 or GIP receptor. (GCNF , TR2 - 110 , ß , TR4 , ERA , B ERRA , ß ) . 60 The activity of the glucagon related peptide at the glucagon Pre- proglucagon is a 158 amino acid precursor polypep - receptor, at the GLP - 1 receptor, or at the GIP receptor can be tide that is processed in different tissues to form a number of in accordance with any of the teachings set forth herein . In different proglucagon - derived peptides, including glucagon , some specific embodiments , the glucagon related peptide glucagon - like peptide - 1 (GLP - 1 ) , glucagon - like peptide - 2 exhibits at least 0 . 1 % activity of native glucagon at the (GLP - 2 ) and oxyntomodulin (OXM ) , that are involved in a 65 glucagon receptor, at least 0 . 1 % activity of native GLP - 1 at wide variety of physiological functions , including glucose the GLP - 1 receptor, or at least 0 . 1 % activity of native GIP homeostasis , insulin secretion , gastric emptying, and intes- at the GIP receptor. US 9 , 783 ,592 B2 The NHR ligand ( Y ) is wholly or partly non - peptidic and levels between days 0 and 14 , demonstrating that the acts at a nuclear hormone receptor with an activity in enhanced ability to improve glucose independent of a accordance with any of the teachings set forth herein . In change in body weight. some embodiments the NHR ligand has an EC se or ICs of FIGS . 3a - 3e illustrate the effect of administration of the about 1 mM or less , or 100 uM or less , or 10 uM or less , or 5 indicated GLP - 1 conjugates on blood glucose, body weight, 1 uM or less . In some embodiments, the NHR ligand has a fat mass , and lean muscle mass in diet induced obese mice . molecular weight of up to about 5000 daltons , or up to about FIG . 3a illustrates the results of a ipGTT test on day 21 . 2000 daltons, or up to about 1000 daltons , or up to about500 FIGS . 3b - d illustrate the effect of administration of the daltons. The NHR ligand may act at any of the nuclear indicated GLP - 1 conjugates on the change in body weight hormone receptors described herein or have any of the 10 (FIG . 3b ), change in fat mass (FIG . 3c ) , and change in lean structures described herein . muscle mass ( FIG . 3d ) in diet induced obese mice . Mice that In some embodiments, the glucagon related peptide has were administered a high dose of the GLP - 1 an EC50 (or IC50 ) at the glucagon receptor within about ( Aib - Aº- CexK + )/ Estrogen ( 17 -ester ) conjugate experienced 100 - fold , or within about 75 - fold , or within about 50 - fold , or the greatest decrease in body weight, and fat mass, and the within about 40 -, 30 -, 25 - , 20 -, 15 ., 10 - or 5 - fold of the EC50 15 least amount of change in lean muscle mass . FIG . 3e or IC50 of the NHR ligand at its nuclear hormone receptor illustrates the effect of administration of the indicated GLP - 1 In some embodiments , the glucagon related peptide has an conjugates on changes in blood glucose in diet induced EC50 ( or IC50 ) at the GLP - 1 receptor within about 100 - fold , obese mice . Mice that were administered a high dose of the or within about 75 - fold , or within about 50 - fold , or within GLP - 1 ( Aib A22CexK49 ) /Estrogen ( 17 - ester ) conjugate about 40 - , 30 -, 25 - , 20 -, 15 -, 10 - or 5 - fold of the EC so or 20 experienced the greatest decrease in blood glucose levels IC50 of the NHR ligand at its nuclear hormone receptor. In between days 0 and 14 . These results demonstrate the added some embodiments , the glucagon related peptide has an dose dependent efficacy of adding estrogen to a weak EC 50 (or IC50 ) at the GIP receptor within about 100 - fold , or A22 - based GLP - 1 agonist . within about 75 - fold , or within about 50 - fold , or within FIGS. 4a -4d illustrate the effect of administration of the about 40 -, 30 -, 25- , 20 -, 15 -, 10 - or 5 - fold of the EC 50 or 25 indicated GLP - 1 conjugates on blood glucose levels , change IC50 of the NHR ligand at its nuclear hormone receptor. in body weight, change in fat mass , and change in blood In some aspects of the invention , prodrugs of O - L - Y are glucose . FIG . 4a illustrates the effect of administration of the provided wherein the prodrug comprises a dipeptide prodrug indicated GLP - 1 conjugates in an intraperitoneal glucose element ( A - B ) covalently linked to an active site of Q via an tolerance test on blood glucose levels in diet induced obese amide linkage . Subsequent removal of the dipeptide under 30 mice after fourteen daily doses . FIGS . 4b - c illustrate the physiological conditions and in the absence of enzymatic effect of administration of the indicated GLP - 1 conjugates activity restores full activity to the Q - L - Y conjugate . on the change in body weight and change in fat mass in diet In some aspects of the invention , pharmaceutical compo - induced obese mice . Mice that were administered a high sitions comprising the Q - L - Y conjugate and a pharmaceu dose of the GLP - 1 / Estrogen conjugates experienced the tically acceptable carrier are also provided . 35 greatest decrease in total body weight (FIG . 46 ) . Fat mass In other aspects of the invention , methods are provided for was decreased in the high dose estrogen ether conjugate administering a therapeutically effective amount of a Q - L - Y relative to vehicle - treated animals (FIG . 40 ). FIG . 4d illus conjugate described herein for treating a disease or medical trates the effect of administration of the indicated GLP - 1 condition in a patient. In some embodiments , the disease or conjugates on changes in blood glucose in diet induced medical condition is selected from the group consisting of 40 obese mice . At the high dose , mice that were administered metabolic syndrome, diabetes , obesity, liver steatosis , and a either GLP - 1 ( Aib ? E Cex K49 ) / Estrogen conjugate experi neurodegenerative disease . enced a greater change in blood glucose levels between days 0 and 21 than mice that were administered GLP - 1 BRIEF DESCRIPTION OF THE DRAWINGS ( Aib ? El Cex K40) alone . 45 FIGS . 5a - 5b illustrate the effect of administration of the FIG . 1 presents an alignment of the amino acid sequences indicated GLP - 1 conjugates on change in body weight and of various glucagon superfamily peptides or relevant frag change in blood glucose in diet induced obese mice . Mice ments thereof. The amino acid sequence presented are that were administered a high dose of the GLP - 1 /Estrogen GHRH (SEQ ID NO : 1619 ) , PHI (SEQ ID NO : 1622 ) , VIP conjugates experienced the greatest decrease in body weight ( SEO ID NO : 1620 ) , PACAP - 27 ( SEO ID NO : 1621) , 50 ( FIG . 5a ) . Mice that were administered either high or low Exendin - 4 (SEQ ID NO : 1618 ) , GLP - 1 (SEQ ID NO : 1603 ) , doses of GLP - 1 ( Aib?E16K4ºCex ) experienced the greatest Glucagon (SEO ID NO : 1601 ) , Oxyntomodulin (SEQ ID change in blood glucose , along with that of the high dose NO : 1606 ), GIP (SEQ ID NO : 1607 ), GLP- 2 (SEQ ID NO : GLP - 1 (Aib²E16K4ºCex )/ estrogen ( 3 -ether ) (FIG . 5b ). 1608 ) and Secretin ( SEQ ID NO : 1624 ) . The alignment FIGS . 6a -6c illustrate the effect of administration of the showshow amino acid positions of glucagon can correspond 55 indicated GLP - 1 conjugates on the change in body weight , to amino acid positions in other glucagon superfamily fat mass , and changes in blood glucose in diet induced obese peptides . mice . Mice that were administered either estrogen conjugate FIGS. 2a - 2c illustrate the effect of administration of the experienced the greatest decrease in totalbody weight (FIG . indicated GLP - 1 conjugates on change in body weight and 6a ) . The analysis of fat mass ( FIG . 6b ) was relatively blood glucose levels in db / db mice . FIG . 2a illustrates that 60 constant with total loss in body weight. Mice that were mice that were administered a high dose of the GLP -1 administered a GLP - 1 (Aib²E16K4ºCex )/ Estrogen (3 - ether ) ( Aib2A22CexK40 / Estrogen ( 17 - ester ) conjugate experienced or a GLP - 1 ( Aib ?ElK4ºCex )/ Estrogen ( 17 - ester ) conjugate a slightly greater decrease in body weight than mice that experienced the greatest change in blood glucose levels were administered GLP - 1 alone , but similar to vehicle . between days 0 and 7 (FIG . 6c ) . FIGS. 2b and 2c show mice that were administered a high 65 FIGS. 7a - 7b illustrate the effect of administration of the dose of the GLP - 1 ( Aib ? A22CexK40 )/ Estrogen ( 17 -ester ) indicated GLP - 1 conjugates on the change in body weight conjugate experienced the greatest decrease in blood glucose and blood glucose in diet induced obese mice. Mice that US 9 ,783 , 592 B2 were administered the GLP - 1 (Aib²E16K4ºCex )/ Estrogen a high dose (FIG . 12a ). FIG . 12b illustrates that mice that conjugates experienced the greatest decrease in total body were administered the inactive , estrogen - labile d -amino acid weight (FIG . 7a ) and blood glucose levels (FIG . 7b ) over a containing GLP - 1 /Estrogen ( 3 -ester ) conjugate consumed period of 7 days. Neither the A22 or the d -amino acid significantly less food than mice that were administered containing peptides demonstrated much lowering relative to 5 inactive d -amino acid containing GLP - 1 alone or an estro the GLP - 1 ( Aib ?El Cex K40 ) /Estrogen conjugates. Addition - gen - stable d -amino acid containing GLP - 1 / Estrogen ( 3 ally , the estrogen conjugates of GLP - 1( Aib²E16Cex K40) ether ) conjugate , with the effect more pronounced at a high were clearly more effective than the non -estrogen form of dose . FIG . 12c shows that a high dose of the estrogen -labile , the same peptide . inactive d - amino acid containing GLP - 1 /Estrogen ( 3 -ester ) FIGS . 8a - 8c illustrate the effect of the administration of 10 conjugate lowers blood glucose levels relative to vehicle the indicated GLP - 1 conjugates on change in body weight, treated animals . FIG . 12d shows that mice that were admin blood glucose, and fat mass . The d -amino acid containing istered the estrogen - labile , inactive d - amino acid containing peptides were clearly inferior in all measures of efficacy to GLP - 1 / Estrogen ( 3 - ether ) conjugate experienced a subtle but the 1 - amino acid containing peptides . FIG . Sa illustrates the greater decrease in liver weight than mice that were admin effect of administration of the indicated GLP - 1 conjugates 15 istered the estrogen -stable , inactive d -amino acid containing on the change body weight in diet induced obese mice . There GLP - 1 / Estrogen ( 3 - ether ) conjugate. FIG . 12e shows that was little apparent difference in body weight lowering at mice that were administered the inactive , estrogen - labile these doses for the peptides with and without the estrogen . d -amino acid containing GLP - 1 /Estrogen ( 3 - ester ) conjugate Nonetheless , in FIG . 8b effect of administration of the experienced a significantly greater increase in uterus weight indicated GLP - 1 conjugates on the change in blood glucose 20 than mice that were administered inactive GLP - 1 alone or in diet induced obese mice are shown . Mice that were the inactive , estrogen - stable d -amino acid containing GLP administered the GLP - 1 ( Aib El Cex K40 )/ Estrogen ( 3 -ester ) 1 /Estrogen ( 3 -ether ) conjugate . conjugate experienced the greatest change in blood glucose FIGS. 13a - 13e illustrate the effect of administration of the levels between days 0 and 7 in vivo , far more than the indicated GLP - 1 conjugates on the change in body weight, animals treated with the same peptide but without estrogen . 25 fat mass , food intake, blood glucose levels , and uterine This demonstrated the direct improvement in blood glucose weight of ovariectomized mice . FIG . 13a shows that mice independent of a difference in body weight. FIG . 8c illus that were administered the active , estrogen -stable GLP - 1 trates the effect of the administration of the indicated GLP - 1 agonist/ Estrogen ( 3 -ether ) conjugate experienced a greater conjugates on change in fat mass . decrease in body weight than mice that were administered FIGS. 9a -9c illustrate the effect of administration of the 30 the active, GLP - 1 agonist alone , the inactive, estrogen -labile indicated GLP - 1 conjugates on the change in body weight, d -amino acid containing GLP - 1 /Estrone ( 3 - ester ) conjugate , amount of fat mass , and change in blood glucose in diet or the inactive , estrogen - stable d -amino acid containing induced obese mice . Mice that were administered the GLP - 1 /Estrogen ( 3 - ether ) conjugate . FIG . 13b shows that d - amino acid containing GLP - 1 /Estrogen ( 3 - ester ) conjugate mice that were administered the GLP - 1 agonist, the inactive , at elevated doses relative to the control 1 - containing amino 35 estrogen - labile d - amino acid containing GLP - 1 / Estrone ( 3 acid experienced the greatest decrease in body weight ( FIG . ester ) conjugate , and the active , estrogen -stable GLP - 1 9a ) and had the least amount of fat mass ( FIG . 96 ) . There agonist /Estrogen ( 3 - ether ) conjugate exhibited a decrease in was clear dose dependent glucose lowering in the d - amino fat mass . FIG . 13c shows that mice that were administered acid containing GLP - 1 / Estrogen ( 3 -ester ) conjugate between the active , estrogen -stable GLP - 1 agonist / Estrogen ( 3 - ether ) days 0 and 7 and enhanced at the highest dose relative to 40 conjugate consumed less food than mice that were admin mice that were administered GLP - 1 alone ( FIG . 9c ) . istered the active , GLP - 1 agonist alone, the inactive , estro FIGS . 10a - 10b illustrate the effect of administration of the gen - labile d - amino acid containing GLP - 1 /Estrone ( 3 - ester ) indicated GLP - 1 conjugates on the change in body weight conjugate , or the inactive , estrogen -stable d - amino acid and change in blood glucose levels in diet induced obese containing GLP - 1 / Estrogen ( 3 - ether ) conjugate . FIG . 13d mice . Mice that were administered the d -amino acid con - 45 shows that mice that were administered the inactive , estro taining GLP - 1 /Estrogen conjugates where the estrogen was gen -labile d -amino acid containing GLP - 1 /Estrone ( 3 - ester ) covalently linked as either a stable amide or ester that was conjugate , or inactive , estrogen -stable d -amino acid contain unstable in vivo . The animals treated with the unstable ester i ng GLP - 1 / Estrogen ( 3 - ether ) conjugate , or the active , estro conjugate experienced the greatest decrease in total body g en - stable GLP - 1 agonist / Estrogen ( 3 - ether ) conjugate expe weight ( FIG . 10a ) . FIG . 10b illustrates that mice adminis - 50 rienced a decrease in blood glucose levels greater than tered the d -amino acid containing GLP - 1 /Estrogen ester vehicle , and that mice that were administered the inactive , conjugate experienced a greater change in blood glucose estrogen -stable d -amino acid containing GLP - 1 / Estrogen ( 3 levels between days 0 and 7 than mice that were adminis - ether ) conjugate did not exhibit a decrease in blood glucose tered a comparable d -amino acid containing peptide butwith levels. FIG . 13e shows that mice that were administered the a stable estrogen conjugate . 55 inactive, estrogen - labile d -amino acid containing GLP - 1 / FIGS. 11a - 11b illustrate the activity of the indicated Estrone ( 3 - ester ) conjugate experienced a significantly conjugates at the GLP - 1 receptor and the estrogen receptor. greater increase in uterus weight than mice that were admin The active , metastable GLP - 1 / estrogen conjugates were istered GLP - 1 alone , the inactive, estrogen - stable d - amino equally active at the GLP - 1 receptor ( FIG . 11a ) , and had acid containing GLP - 1 /Estrogen ( 3 - ether ) conjugate , or the variable activity at the estrogen receptor ( FIG . 11b ). 60 active , estrogen -stable GLP - 1 /Estrogen ( 3 -ether ) conjugate . FIGS . 12a - 12e illustrate the effect of administration of the FIGS. 14a - 14d illustrate the effect of administration of the indicated GLP - 1 conjugates on the change in body weight, indicated GLP - 1 conjugates on the change in body weight, food intake , blood glucose , liver weight, and uterus weight fat mass , blood glucose , and uterine weight of ovariecto in diet induced obese mice . Mice that were administered an mized mice . Mice that were administered the metastable inactive, estrogen - labile d - amino acid containing GLP- 1 / 65 GLP - 1 agonist /Estrogen conjugates experienced a greater Estrogen ( 3 - ester ) conjugate experienced the greatest decrease in body weight and fat mass than mice that were decrease in body weight, with the effect more pronounced at administered the active, GLP- 1 agonist alone or the active, US 9 ,783 ,592 B2 estrogen - labile GLP - 1 agonist /Estrone ( 3 - ester) conjugate The term “ about” as used herein means greater or lesser ( FIGS . 14a and 14b ) . Mice that were administered the than the value or range of values stated by 10 percent, but metastable enzyme- and acid - labile conjugates , GLP - 1 ago - is not intended to designate any value or range of values to nist/ Estrogen ( 17 -cathepsin ) and GLP - 1 agonist / Estrogen only this broader definition . Each value or range of values ( 17 -hydrazone ), respectively, initially had the greatest 5 preceded by the term " about” is also intended to encompass decrease in body weight, while the metastable thiol reduc the embodiment of the stated absolute value or range of tion - labile conjugate , GLP - 1 agonist/ Estrogen ( 17 - carbam values. ate disulfide ), exhibited the overall greatest decrease in body As used herein , the term “ pharmaceutically acceptable weight. FIG . 14c shows thatmice that were administered the metastable thiol reduction - and acid - labile conjugates , 10 carrier” includes any of the standard pharmaceutical carri GLP - 1 agonist/ Estrogen ( 17 - carbamate disulfide ) and GLP - 1 ers , such as a phosphate buffered saline solution , water , agonist/ Estrogen ( 17 -hydrazone ) , respectively , experienced emulsions such as an oil/ water or water / oil emulsion , and a greater decrease in blood glucose levels than the estrogen various types of wetting agents . The term also encompasses labile GLP - 1 agonist/ Estrone ( 3 - ester ) conjugate . FIG . 14d any of the agents approved by a regulatory agency of the US shows that mice that were administered the estrogen - labile 15 Federal government or listed in the US Pharmacopeia for GLP - 1 /Estrone ( 3 -ester ) conjugate experienced a signifi use in animals , including humans. cantly greater increase in uterus weight than mice that were As used herein the term “ pharmaceutically acceptable administered any of the three metastable GLP - 1 /Estrogen salt” refers to salts of compounds that retain the biological conjugates . activity of the parent compound , and which are not biologi FIGS . 15a - 15b illustrate the effect of administration of the 20 cally or otherwise undesirable . Many of the compounds indicated GLP -1 conjugates on the change in body weight disclosed herein are capable of forming acid and /or base and cumulative food intake . Mice that were administered the salts by virtue of the presence of amino and /or carboxyl active , estrogen -stable GLP - 1 agonist/ Estrogen ( 3 -ether groups or groups similar thereto . conjugate experienced a significantly greater decrease in Pharmaceutically acceptable base addition salts can be body weight than mice that were administered the active , 25 prepared from inorganic and organic bases . Salts derived GLP - 1 agonist alone or the active , estrogen - labile GLP - 1 from inorganic bases, include by way of example only , agonist/ Estrogen (3 -ester ) conjugate (FIG . 15a ). FIG . 15b sodium , potassium , lithium , ammonium , calcium and mag shows that mice that were administered the active , estrogen nesium salts . Salts derived from organic bases include , but stable GLP - 1 agonist / Estrogen ( 3 - ether ) conjugate con - are not limited to , salts of primary , secondary and tertiary sumed significantly less food than mice that were adminis - 30 amines. Pharmaceutically acceptable acid addition salts may tered the active , GLP - 1 agonist alone or the active , estrogen - be prepared from inorganic and organic acids . Salts derived labile GLP - 1 agonist /Estrogen (3 -ester ) conjugate . from inorganic acids include hydrochloric acid , hydrobro FIGS . 160 - 16e depict HPLC profiles that illustrate the mic acid , sulfuric acid , nitric acid , phosphoric acid , and the stability of the indicated GLP - 1 /Estrogen conjugates in like . Salts derived from organic acids include acetic acid , human plasma at 37° C . The estrogen - stable GLP - 1 / Estro - 35 propionic acid , glycolic acid , pyruvic acid , oxalic acid , gen ( 3 - ether ) conjugates exhibited no estrogen release over malic acid , malonic acid , succinic acid , maleic acid , fumaric 72 hours , while the estrogen - labile GLP - 1 /Estrogen ( 3 -ester ) acid , tartaric acid , citric acid , benzoic acid , cinnamic acid , conjugates showed substantial estrogen release after 3 hours mandelic acid , methanesulfonic acid , ethanesulfonic acid , and complete estrogen release within 6 hours . p - toluene - sulfonic acid , salicylic acid , and the like . FIGS . 17a - 17c illustrate the effect of administration of the 40 As used herein , the term " treating ” includes prophylaxis indicated conjugates on the change in body weight percent of the specific disorder or condition , or alleviation of the age, cumulative food intake , and blood glucose levels of symptoms associated with a specific disorder or condition diet - induced obesity mice . and /or preventing or eliminating said symptoms. For FIGS . 18a - 18c illustrate the effect of administration of the example , as used herein the term “ treating diabetes” will indicated conjugates on the change in body weight percent- 45 refer in general to altering glucose blood levels in the age , cumulative food intake , and blood glucose levels on direction of normal levels and may include increasing or diet -induced obesity wild type mice , estrogen receptor beta decreasing blood glucose levels depending on a given situ knock -out (ERPB KO ) mice , and estrogen receptor alpha ation . knock -out (ERa KO ) mice . As used herein an " effective” amount or a “ therapeutically FIG . 19 illustrates the effect of the indicated GLP - 1 50 effective amount” of a glucagon peptide refers to a nontoxic conjugates on blood glucose levels with time. Mice that but sufficient amount of the peptide to provide the desired were administered the GLP - 1 agonist showed the least effect . For example one desired effect would be the preven effective lowering of blood glucose over 48 hours ( except tion or treatment of hypoglycemia , as measured , for for vehicle ) , while mice that were administered the estrogen - example , by an increase in blood glucose level . An alterna stable GLP - 1 agonist/ Estrogen ( 3 - ether ) conjugate showed 55 tive desired effect for the glucagon peptides of the present the most effective decrease in blood glucose over 48 hours . disclosure would include treating hyperglycemia , e . g ., as FIGS . 20A , 20B and 20C provide the structures of various measured by a change in blood glucose level closer to embodiments of linkers used in accordance the present normal, or inducing weight loss /preventing weight gain , disclosure . e . g . , as measured by reduction in body weight, or preventing 60 or reducing an increase in body weight , or normalizing body DETAILED DESCRIPTION OF THE fat distribution . The amount that is " effective” will vary INVENTION from subject to subject , depending on the age and general condition of the individual, mode of administration , and the Definitions like . Thus , it is not always possible to specify an exact In describing and claiming the invention , the following 65 “ effective amount. ” However, an appropriate “ effective ” terminology will be used in accordance with the definitions amount in any individual case may be determined by one of set forth below . ordinary skill in the art using routine experimentation . US 9 , 783 ,592 B2 10 The term , “ parenteral ” means not through the alimentary (iii ) a deletion of one or more amino acids from the reference canal but by some other route, e . g . , subcutaneous , intramus - peptide ( e . g . SEQ ID NOs: 1601, 1603 , 1607 ) . cular, intraspinal, or intravenous . In some embodiments , the amino acid substitution or As used herein the term “ patient” without further desig - replacement is a conservative amino acid substitution , e . g. , nation is intended to encompass any warm blooded verte - 5 a conservative substitution of the amino acid at one or more brate domesticated animal (including for example , but not of positions 1 , 2 , 5 , 7 , 8 , 10 , 11, 12 , 13 , 14 , 16 , 17 , 18 , 19 , 20 , 21 , 24 , 27 , 28 or 29 . As used herein , the term " conser limited to livestock , horses, cats , dogs and other pets ), vative amino acid substitution ” is the replacement of one mammals , and humans . amino acid with another amino acid having similar proper The term “ isolated ” as used herein means having beendi 10 ties , e. g. , size , charge, hydrophobicity , hydrophilicity , and / or removed from its natural environment. In some embodi- ? aromaticity , and includes exchanges within one of the fol ments, the analog is made through recombinant methods and lowing five groups: the analog is isolated from the host cell . I . Small aliphatic , nonpolar or slightly polar residues: The term “ purified , ” as used herein relates to the isolation Ala , Ser , Thr, Pro , Gly ; of a molecule or compound in a form that is substantially 15 II. Polar, negative- charged residues and their amides and free of contaminants normally associated with the molecule esters : or compound in a native or natural environment and means Asp , Asn , Glu , Gin , cysteic acid and homocysteic acid ; having been increased in purity as a result of being separated III. Polar, positive - charged residues : from other components of the original composition . The His, Arg , Lys ; Ornithine (Orn ) term “ purified polypeptide” is used herein to describe a 20 IV . Large, aliphatic , nonpolar residues: polypeptide which has been separated from other com Met , Leu , Ile , Val , Cys, Norleucine (Nle ) , homocysteine pounds including , but not limited to nucleic acid molecules , V . Large , aromatic residues : lipids and carbohydrates. Phe , Tyr, Trp , acetyl phenylalanine As used herein , the term " peptide ” encompasses a In some embodiments , the amino acid substitution is not sequence of 2 or more amino acids and typically less than 50 25 a conservative amino acid substitution , e . g ., is a non amino acids , wherein the amino acids are naturally occurring conservative amino acid substitution . or coded or non -naturally occurring or non - coded amino As used herein the term “ amino acid ” encompasses any acids . Non - naturally occurring amino acids refer to amino molecule containing both amino and carboxyl functional acids that do not naturally occur in vivo but which , never groups , wherein the amino and carboxyl groups are attached theless , can be incorporated into the peptide structures 30 to the same carbon ( the alpha carbon ) . The alpha carbon described herein . “ Non - coded ” as used herein refer to an optionally may have one or two further organic substituents . amino acid that is not an L - isomer of any of the following for the purposes of the present disclosure designation of an 20 amino acids : Ala , Cys, Asp , Glu , Phe, Gly , His , Ile , Lys, amino acid without specifying its stereochemistry is Leu , Met , Asn , Pro , Gln , Arg , Ser, Thr, Val, Trp , Tyr. intended to encompass either the L or D form of the amino As used herein , " partly non - peptidic ” refers to a molecule 35 acid , or a racemic mixture . However , in the instance where wherein a portion of the molecule is a chemical compound an amino acid is designated by its three letter code and or substituent that has biological activity and that does not includes a superscript number (i . e ., Lys = ), such a designa comprises a sequence of amino acids . tion is intended to specify the native L form of the amino As used herein , " non -peptidic ” refers to a molecule has acid , whereas the D form will be specified by inclusion of a biological activity and that does not comprise a sequence of 40 lower case d before the three letter code and superscript amino acids. number ( i . e . , dLys - 1 ) . As used herein , the terms " polypeptide ” and “ protein ” are As used herein the term “ hydroxyl acid ” refers to an terms that are used interchangeably to refer to a polymer of amino acid that has been modified to replace the alpha amino acids, without regard to the length of the polymer . carbon amino group with a hydroxyl group . Typically , polypeptides and proteins have a polymer length 45 As used herein the term “ charged amino acid ” refers to an that is greater than that of " peptides. " In some instances, a amino acid that comprises a side chain that has a negative protein comprises more than one polypeptide chain cova - charge ( i . e . , deprotonated ) or positive charge ( i. e ., proto lently or noncovalently attached to each other . nated ) in aqueous solution at physiological pH . For example Throughout the application , all references to a particular negative - charged amino acids include aspartic acid , gluta amino acid position by number ( e . g . , position 28 ) refer to the 50 mic acid , cysteic acid , homocysteic acid , and homoglutamic amino acid at that position in native glucagon (SEQ ID NO : acid , whereas positive -charged amino acids include argi 1601 ) or the corresponding amino acid position in any nine , lysine and histidine . Charged amino acids include the analogs thereof. For example , a reference herein to " position charged amino acids among the 20 coded amino acids, as 28 ” would mean the corresponding position 27 for an analog well as atypical or non -naturally occurring or non - coded of glucagon in which the first amino acid of SEQ ID NO : 55 amino acids . 1601 has been deleted . Similarly , a reference herein to As used herein the term “ acidic amino acid ” refers to an " position 28 ” would mean the corresponding position 29 for amino acid that comprises a second acidic moiety (other a analog of glucagon in which one amino acid has been than the alpha carboxylic acid of the amino acid ) , including added before the N - terminus of SEQ ID NO : 1601. for example , a side chain carboxylic acid or sulfonic acid As used herein an " amino acid modification ” refers to ( i ) 60 group . a substitution or replacement of an amino acid of the As used herein an “ acylated ” amino acid is an amino acid reference peptide ( e . g . SEQ ID NOs: 1601 , 1603, 1607 ) with comprising an acyl group which is non -native to a naturally a different amino acid (naturally occurring or coded or occurring amino acid , regardless by the means by which it non -coded or non - naturally occurring amino acid ) , ( ii ) an is produced . Exemplary methods of producing acylated addition of an amino acid (naturally - occurring or coded or 65 amino acids and acylated peptides are known in the art and non -coded or non -naturally -occurring amino acid ) to the include acylating an amino acid before inclusion in the reference peptide ( e .g . SEQ ID NOs: 1601 , 1603 , 1607 ) or peptide or peptide synthesis followed by chemical acylation US 9 ,783 , 592 B2 12 of the peptide. In some embodiments , the acyl group causes enyl, OCF3, NO2, CN , NC , OH , alkoxy, amino , CO , H , the peptide to have one or more of (i ) a prolonged half- life C3 - C , cycloalkyl, C ( O )Oalkyl , aryl, and heteroaryl. Exem in circulation , ( ii ) a delayed onset of action , ( iii ) an extended plary aryl groups include , but are not limited to , phenyl , duration of action , ( iv ) an improved resistance to proteases , naphthyl, tetrahydronaphthyl, chlorophenyl, indanyl , inde such as DPP - IV , and ( v ) increased potency at the glucagon 5 nyl, methylphenyl, methoxyphenyl, trifluoromethylphenyl, superfamily peptide receptor. nitrophenyl, 2 , 4 -methoxychlorophenyl , and the like . As used herein , an " alkylated ” amino acid is an amino As used herein , the term " heteroaryl” refers to a mono acid comprising an alkyl group which is non -native to a cyclic or polycyclic ring system containing one or more naturally occurring amino acid , regardless of the means by aromatic rings and containing at least one nitrogen , oxygen , which it is produced . Exemplary methods of producing 10 or sulfur atom in an aromatic ring. The size of the heteroaryl alkylated amino acids and alkylated peptides are known in ring and the presence of substituents or linking groups are the art and including alkylating an amino acid before inclu - indicated by designating the number of carbons present. For sion in the peptide or peptide synthesis followed by chemi- example , the term “ (C1 - C6 alkyl) (C5 - C6 heteroaryl) ” refers cal alkylation of the peptide . Without being held to any to a 5 or 6 membered heteroaryl that is attached to a parent particular theory, it is believed that alkylation of peptides 15 moiety via a one to 6 membered alkyl chain . Unless other will achieve similar, if not the same, effects as acylation of wise indicated , a heteroaryl group can be unsubstituted or the peptides, e . g . , a prolonged half- life in circulation , a substituted with one or more , and in particular one to five delayed onset of action , an extended duration of action , an groups independently selected from , for example , halo , improved resistance to proteases , such as DPP - IV , and alkyl, alkenyl, OCF , NO , , CN , NC , OH , alkoxy , amino , increased potency at the glucagon superfamily peptide 20 COQH , Cz- Cg cycloalkyl , C ( O )Oalkyl , aryl, and heteroaryl . rereceptor . Examples of heteroaryl groups include , but are not limited The term “ C , -C , alkyl” wherein n can be from 1 through to , thienyl , furyl, pyridyl, oxazolyl, quinolyl, thiophenyl, 18 , as used herein , represents a branched or linear alkyl isoquinolyl , indolyl, triazinyl, triazolyl, isothiazolyl, isox group having from one to the specified number of carbon azolyl, imidazolyl, benzothiazolyl, pyrazinyl, pyrimidinyl, atoms. For example , C . - C . alkyl represents a branched or 25 thiazolyl, and thiadiazolyl. linear alkyl group having from 1 to 6 carbon atoms. Typical As used herein , the term " heteroalkyl” refers to a linear or C - Cis alkyl groups include, but are not limited to , methyl, branched hydrocarbon containing the indicated number of ethyl, n -propyl , iso -propyl , butyl , iso -butyl , sec -butyl , tert - carbon atoms and at least one heteroatom in the backbone of butyl , pentyl, hexyl and the like . Alkyl groups optionally can the structure . Suitable heteroatoms for purposes herein be substituted , for example , with hydroxy (OH ) , halo , aryl, 30 include but are not limited to N , S , and O . Heteroalkyl carboxyl , thio , Cz- C , cycloalkyl, and amino . groups optionally can be substituted , for example , with The term " Co- C , alkyl” wherein n can be from 1 - 18 , as hydroxy (OH ), halo , aryl, carboxyl, and amino . used herein , represents a branched or linear alkyl group As used herein , the term “ halogen ” or “ halo ” refers to one having up to 18 carbon atoms. For example , the term or more members of the group consisting of fluorine, chlo " (Co -Co alkyl )OH ” represents a hydroxyl parent moiety 35 rine , bromine , and iodine. attached to an alkyl substituent having up to 6 carbon atoms As used herein , the term “ glucagon related peptide ” refers ( e . g . OH , CH2OH , C2H4OH , CZHGOH , to those peptides which have biological activity ( as agonists - C4H , OH , C3H10OH , C6H 2OH ) . or antagonists ) at any one or more of the glucagon , GLP - 1 , The term " C2- Cn alkenyl” wherein n can be from 2 GLP - 2, and GIP receptors and comprise an amino acid through 18 , as used herein , represents an unsaturated 40 sequence that shares at least 40 % sequence identity ( e . g . , branched or linear group having from 2 to the specified 45 % , 50 % , 55 % , 60 % , 65 % , 70 % , 75 % , 80 % , 85 % , 90 % , number of carbon atoms and at least one double bond . 95 % ) with at least one of native glucagon , native oxynto Examples of such groups include , but are not limited to , modulin , native exendin - 4 , native GLP - 1, native GLP - 2, or 1 -propenyl , 2 -propenyl ( CH2CH = CH2 ) , 1, 3 -butadi native GIP . Unless otherwise stated , any reference to an ACH = CHCH = CH2) , 1 -butenyl 45 amino acid position in a glucagon related peptide ( e . g . for ICH = CHCH2CH3) , hexenyl , pentenyl, and the like . Alk - linkage of a NHR ligand , a conjugate moiety , a hydrophilic enyl groups optionally can be substituted , for example , with polymer , acylation or alkylation ) refers to the corresponding hydroxy (OH ) , halo , aryl , carboxyl, thio , C3 - C , cycloalkyl, position relative to the native glucagon amino acid sequence and amino . (SEO ID NO : 1601 ) . The term " C2- C , alkynyl” wherein n can be from 2 to 18 , 50 As used herein , the term “ selectivity ” of a molecule for a refers to an unsaturated branched or linear group having first receptor relative to a second receptor refers to the from 2 to n carbon atoms and at least one triple bond . following ratio : EC50 of the molecule at the second receptor Examples of such groups include , but are not limited to , divided by the EC 50 of themolecule at the first receptor . For 1 - propynyl, 2 -propynyl , 1 -butynyl , 2 -butynyl , 1 -pentynyl , example , a molecule that has an EC50 of 1 nM at a first and the like . Alkynyl groups optionally can be substituted , 55 receptor and an EC50 of 100 nM at a second receptor has for example , with hydroxy (OH ) , halo , aryl, carboxyl, thio , 100 - fold selectivity for the first receptor relative to the Cz- C , cycloalkyl, and amino . second receptor. As used herein the term “ aryl” refers to a monocyclic or The term “ identity ” as used herein relates to the similarity polycyclic ( for example , bicyclic , tricyclic , or tetracyclic ) between two or more sequences. Identity is measured by aromatic groups. The size of the aryl ring or rings is 60 dividing the number of identical residues by the total num indicated by designating the number of carbons present . For ber of residues and multiplying the product by 100 to example , the term “ ( C , -Cz alkyl) (C6 - C10 aryl) ” refers to a 6 achieve a percentage . Thus, two copies of exactly the same to 10 membered aryl that is attached to a parent moiety via sequence have 100 % identity , whereas two sequences that a one to three membered alkyl chain . Unless otherwise have amino acid deletions , additions , or substitutions rela indicated , an aryl group can be unsubstituted or substituted 65 tive to one another have a lower degree of identity . Those with one or more , and in particular one to five groups skilled in the art will recognize that several computer independently selected from , for example, halo , alkyl, alk programs, such as those that employ algorithms such as US 9 ,783 ,592 B2 13 14 BLAST ( Basic Local Alignment Search Tool, Altschul et al . GLP - 1 receptor. Also , the term “ GIP agonist peptide ” should ( 1993 ) J . Mol. Biol. 215 :403 - 410 ) are available for deter not be construed as limiting the compound to only peptides . mining sequence identity . Rather, compounds other than peptides are encompassed by As used herein , the term “ glucagon superfamily peptide ” this term . Accordingly , the GIP agonist peptide in some refers to a group of peptides related in structure in their 5 aspects is a peptide in conjugate form ( a heterodimer, a N - terminal and C - terminal regions ( see , for example , Sher - multimer, a fusion peptide ) , a chemically - derivatized pep wood et al ., Endocrine Reviews 21: 619 -670 (2000 ) ) . Mem - tide , a pharmaceutical salt of a peptide , a peptidomimetic , bers of this group include all glucagon related peptides , as and the like . well as Growth Hormone Releasing Hormone (GHRH ; SEQ T he term " glucagon antagonist peptide ” refers to a com ID NO : 1619 ), vasoactive intestinal peptide (VIP ; SEQ ID 10 pound that counteracts glucagon activity or prevents gluca NO : 1620 ) , pituitary adenylate cyclase -activating polypep - gon function . For example , a glucagon antagonist exhibits at tide 27 ( PACAP - 27 ; SEQ ID NO : 1621) , peptide histidine least 60 % inhibition ( e . g ., at least 70 % , 80 % , 90 % or more isoleucine (PHI ; SEQ ID NO : 1642 ), peptide histidine inhibition ) of the maximum response achieved by glucagon methionine ( PHM ; SEQ ID NO : 1622 ) , Secretin (SEQ ID at the glucagon receptor. In a specific embodiment, the NO : 1623 ) , and analogs, derivatives or conjugates with up to 15 glucagon antagonist at a concentration of about 1 UM 1 , 2 , 3 , 4 , 5 , 6 , 7 , 8 , 9 or 10 amino acid modifications relative exhibits less than about 20 % of the maximum agonist to the native peptide . Such peptides preferably retain the activity achieved by glucagon at the glucagon receptor ( e . g . ability to interact ( agonist or antagonist) with receptors of less than about 10 % or 5 % ) . This term should not be the glucagon receptor superfamily . Unless otherwise stated , construed as limiting the compound to having activity at any reference to an amino acid position in a glucagon 20 only the glucagon receptor. Rather, the glucagon antagonist superfamily peptide ( e . g . for linkage of a NHR ligand , a peptides of the present disclosures may exhibit additional conjugate moiety , a hydrophilic polymer , acylation or alky - activities at the glucagon receptor ( e .g ., partial agonism ) or lation ) refers to the corresponding position relative to the other receptor. Glucagon antagonist peptides , for example , native glucagon amino acid sequence (SEQ ID NO : 1601 ) , may exhibit activity ( e . g . , agonist activity ) at the GLP - 1 see FIG . 1 for an alignment of representative glucagon 25 receptor. Also , the term " glucagon antagonist peptide ” superfamily peptides. should not be construed as limiting the compound to only The term " glucagon agonist peptide ” refers to a com - peptides . Rather, compounds other than peptides are encom pound that binds to and activates downstream signaling of passed by these terms. Accordingly , in some aspects, the the glucagon receptor. However, this term should not be glucagon agonist peptide is a peptide in conjugate form , a construed as limiting the compound to having activity at 30 chemically - derivatized peptide , a pharmaceutical salt of a only the glucagon receptor . Rather , the glucagon agonist peptide , a peptidomimetic , and the like. peptides of the present disclosures may exhibit additional The term “ GLP - 1 antagonist peptide ” refers to a com activities at other receptors, as further discussed herein . pound that counteracts GLP - 1 activity or prevents GLP - 1 Glucagon agonist peptides, for example, may exhibit activ - function . For example , a GLP - 1 antagonist exhibits at least ity ( e . g ., agonist activity ) at the GLP - 1 receptor and/ or the 35 60 % inhibition ( e . g . , at least 70 % , 80 % , 90 % or more GIP receptor. Also , the term “ glucagon agonist peptide ” inhibition ) of the maximum response achieved by GLP - 1 at should not be construed as limiting the compound to only the GLP - 1 receptor. In a specific embodiment, a GLP - 1 peptides . Rather , compounds other than peptides are encom - antagonist at a concentration of about 1 uM exhibits less passed by this term . Accordingly , the glucagon agonist than about 20 % of the maximum agonist activity achieved peptide in some aspects is a peptide in conjugate form ( a 40 by GLP - 1 at the GLP - 1 receptor ( e . g . less than about 10 % heterodimer , a multimer, a fusion peptide ) , a chemically - or 5 % ) . The term should not be construed as limiting the derivatized peptide, a pharmaceutical salt of a peptide , a compound to having activity at only the GLP - 1 receptor. peptidomimetic , and the like . Rather, the GLP - 1 antagonist peptides of the present disclo The term “ GLP - 1 agonist peptide” refers to a compound sures may exhibit additional activities at the GLP - 1 receptor that binds to and activates downstream signaling of the 45 ( e . g ., partial agonism ) or other receptor. GLP - 1 antagonist GLP - 1 receptor. However , this term should not be construed peptides , for example , may exhibit activity ( e . g . , agonist as limiting the compound to having activity at only the activity ) at the glucagon receptor. Also , the term "GLP - 1 GLP - 1 receptor. Rather, the GLP - 1 agonist peptides of the antagonist peptide " should not be construed as limiting the present disclosures may exhibit additional activities at other compound to only peptides . Rather, compounds other than receptors, as further discussed herein . GLP - 1 agonist pep - 50 peptides are encompassed by these terms. Accordingly , in tides , for example , may exhibit activity ( e . g . , agonist activ - some aspects , the GLP - 1 agonist peptide is a peptide in ity ) at the glucagon receptor and / or the GIP receptor. Also , conjugate form , a chemically -derivatized peptide, a phar the term “ GLP - 1 agonist peptide” should not be construed as maceutical salt of a peptide , a peptidomimetic , and the like . limiting the compound to only peptides . Rather, compounds The term “ GIP antagonist peptide ” refers to a compound other than peptides are encompassed by this term . Accord - 55 that counteracts GIP activity or prevents GIP - 1 function . For ingly , the GLP - 1 agonist peptide in some aspects is a peptide example , a GIP antagonist exhibits at least 60 % inhibition in conjugate form (a heterodimer , a multimer, a fusion ( e . g ., at least 70 % , 80 % , 90 % or more inhibition ) of the peptide) , a chemically -derivatized peptide , a pharmaceutical maximum response achieved by GIP at the GIP receptor. In salt of a peptide , a peptidomimetic , and the like . a specific embodiment, a GIP antagonist at a concentration The term “ GIP agonist peptide ” refers to a compound that 60 of about 1 uM exhibits less than about 20 % of the maximum binds to and activates downstream signaling of the GIP agonist activity achieved by GIP at the GIP receptor ( e . g . receptor. However, this term should not be construed as less than about 10 % or 5 % ) . The term should not be limiting the compound to having activity at only the GIP construed as limiting the compound to having activity at receptor. Rather, the GIP agonist peptides of the present only the GIP receptor. Rather , the GIP antagonist peptides of disclosures may exhibit additional activities at other recep - 65 the present disclosures may exhibit additional activities at tors, as further discussed herein . GIP agonist peptides , for the GIP receptor ( e . g . , partial agonism ) or other receptor. example , may exhibit activity ( e . g ., agonist activity ) at the GIP antagonist peptides, for example , may exhibit activity US 9 , 783 ,592 B2 15 16 ( e . g . , agonist activity ) at the glucagon receptor . Also , the hormone receptor; R3 and R * are independently moieties that term “ GIP antagonist peptide ” should not be construed as permit or promote agonist or antagonist activity upon bind limiting the compound to only peptides . Rather, compounds ing of the compound of Formula A to a nuclear hormone other than peptides are encompassed by these terms. receptor, and each dashed line represents an optional double Accordingly , in some aspects , the GIP agonist peptide is a 5 bond . Formula A may further comprise one or more sub peptide in conjugate form , a chemically -derivatized peptide, stituents at one or more of positions 1, 2 , 3 , 4 , 5, 6 , 7, 8 , 9 , a pharmaceutical salt of a peptide , a peptidomimetic , and the 11, 12 , 14 , 15 , 16 , and 17 . Contemplated optional substitu like . As used herein , the terms " glucagon analog ” and “ gluca ents include , but are not limited to , OH , NH , ketone , and gon peptide ” can be used interchangeably to refer to an 10 C7 -C18 alkyl groups . Specific , nonlimiting examples of analog of glucagon that has the indicated activity at a steroids and derivatives thereof include cholesterol, cholic glucagon related peptide receptor. acid estradiol, testosterone, and hydrocortisone . As used herein the term “ native glucagon ” refers to a As used herein , “ bile acids and derivatives thereof” refers peptide consisting of the sequence of SEQ ID NO : 1601 . to compounds, either naturally occurring or synthesized , of As used herein , the term “ native GLP - 1 ” is a generic term 15 Formula M : that designates GLP- 1 (7 -36 ) amide ( SEQ ID NO : 1603 ), GLP - 1 ( 7 - 37 ) acid (SEQ ID NO : 1604 ) or a mixture of those two compounds. Formula M As used herein , the term “ native GIP ” refers to a peptide consisting of SEQ ID NO : 1607 . 20 eliin. As used herein , “ glucagon potency ” or “ potency com Inn Me pared to native glucagon ” of a molecule refers to the ratio of R17 the EC50 of the molecule at the glucagon receptor divided by Me the EC50 of native glucagon at glucagon receptor. As used herein , “ GLP - 1 potency ” or “ potency compared 25 to native GLP - 1 ” of a molecule refers to the ratio of the EC50 of the molecule at GLP - 1 receptor divided by the EC 50 of native GLP - 1 at GLP - 1 receptor. As used herein , “ GIP potency ” or “ potency compared to native GIP ” of a molecule refers to the ratio of the EC 50 of 30 wherein each of R15 R16 and R17 are independently moi the molecule at the GIP receptor divided by the EC50 of eties that permit or promote agonist or antagonist activity native GIP at the GIP receptor. As used herein , "NHR ligand ” refers to a hydrophobic or upon binding of the compound of Formula M to a nuclear lipophilic moiety that has biological activity ( either agonist hormone receptor. In some embodiments , each of R15 and or antagonist ) at a nuclear hormone receptor (NHR ) . The 35 R16 are independently hydrogen , (Co -Cz alkyl) halo , C7 -C18 NHR ligand is wholly or partly non - peptidic . In some alkyl, C2 -C18 alkenyl, C2 -C18 alkynyl, heteroalkyl, or (Co embodiments , the NHR ligand is an agonist that binds to and Cg alkyl )OH ; and R17 is OH , (Co -Cg alkyl) NH (C , -C4 alkyl) activates the NHR . In other embodiments , the NHR ligand SO3H , or (Co - Cg alkyl) NH (C1 - C4 alkyl) COOH . Formula M is an antagonist . In some embodiments , the NHR ligand is may further comprise one or more substituents at one or an antagonist that acts by wholly or partially blocking 40 more of positions 1 , 2 , 3 , 4 , 5 , 6 , 7 , 8 , 9 , 11 , 12 , 14 , 15 , 16 , binding of native ligand to the active site. In other embodi and 17 . Nonlimiting examples of bile acids include cholic ments , the NHR ligand is an antagonist that acts by binding acid , deoxycholic acid , lithocholic acid , chenodeoxycholic to the active site or an allosteric site and preventing activa - acid , taurocolic acid , and glycocholic acid . tion of, or de - activating , the NHR . As used herein , “ cholesterol and derivatives thereof" As used herein , “ nuclear hormone receptors ” (NHRs ) 45 refers to compounds , either naturally occurring or synthe refers to ligand -activated proteins that regulate gene expres - sized , comprising a structure similar to that of cholesterol, as sion within the cell nucleus, sometimes in concert with other shown below : co -activators and co - repressors. As used herein , “ steroids and derivatives thereof” refers to compounds, either naturally occurring or synthesized , hav - 50 Melino. ing a structure of Formula A : Me Me.

Formula A Me 55 12 Me R4

Cholesterol 60 7 Derivatives of cholesterol can include oxysterols , such as (R40V 4 66 hydroxycholesterol, 24 ( S )- hydroxycholesterol , 27 -hydroxy cholesterol, and cholestenoic acid . wherein Rl and R , when present, are independently moi- 65 As used herein , “ estradiol and derivatives thereof” refers eties that permit or promote agonist or antagonist activity to compounds, either naturally occurring or synthesized , of upon binding of the compound of Formula A to a nuclear Formula B : US 9 ,783 ,592 B2 17 18 positions of the tetracyclic ring, such as, for example , Formula B positions 1 , 2 , 3 , 4 , 5 , 6 , 7 , 8 , 9 , 11, 12 , 14 , 15 , 16 , and 17 . Specific, nonlimiting examples of derivatives of testosterone Me 0- R include dehydroepiandrosterone, androstenedione , 5 -an URS 5 drostenediol, androsterone , and dihydrotestosterone. As used herein , “ fatty acids and derivatives thereof” refers to carboxylic acids comprising a long unbranched C - to C28 alkyl or C2 to C28 alkenyl moiety and can optionally com R prise one or more halo substituents and / or optionally com 10 prise one or more substituents other than halo . In some embodiments , the long unbranched alkyl or alkenyl moiety wherein R ', R and Roare moieties that permit or promote can bebe wholly halo substitutedsubstitute ( e . g . , all hydrogens replaced agonist or antagonist activity upon binding of the compound with halo atoms) . A short chain fatty acid comprises 1 -5 of Formula B to the estrogen receptor. In some embodi carbon atoms. A medium chain fatty acid comprises 6 - 12 ments , the structure of Formula B is substituted with one or 15 carbon . A long chain fatty acid comprises 13 - 22 carbon more substituents at one or more positions of the tetracyclic atoms. A very long chain fatty acid comprises 23 - 28 carbon ring , such as , for example, positions 1 , 2 , 4 , 6 , 7 , 8 , 9 , 11 , 12 , atoms. Specific , nonlimiting examples of fatty acids include formic acid , acetic acid , n - caproic acid , heptanoic acid , 14 , 15 , 16 , and 17. In some cases , the substituent comprises caprylic acid , nonanoic acid , capric acid , undecanoic acid , a ketone at position -6 . 20 lauric acid , tridecanoic acid , myristic acid , pentadeconoic acid , palmitic acid , heptadecanoic acid , stearic acid , non OH . adecanoic acid , arachidic acid , heneicosanoic acid , behenic Me acid , tricosanoic acid , mead acid , myristoleic acid , palmi toleic acid , sapienic acid , oleic acid , linoleic acid , a - lino lenic acid , elaidic acid , petroselinic acid , arachidonic acid , dihydroxyeicosatetraenoic acid ( DIHETE ) , octadecynoic acid , eicosatriynoic acid , eicosadienoic acid , eicosatrienoic acid , eicosapentaenoic acid , erucic acid , dihomolinolenic HO acid , docosatrienoic acid , docosapentaenoic acid , docosa hexaenoic acid , and adrenic acid . Estradiol As used herein , “ cortisol and derivatives thereof” refers to compounds, either naturally occurring or synthesized , of Specific , nonlimiting examples of derivatives of estradiol Formula C : include B - estradiol 17 - acetate , B - estradiol 17 - cypionate , B - estradiol 17 - enanthate, B - estradiol 17 -valerate , ß - estradiol 35 3 , 17 - diacetate , B - estradiol 3 , 17 -dipropionate , B - estradiol Formula C 3 -benzoate , B - estradiol 3 -benzoate 17 - n -butyrate , ß - estra ROC diol 3 - glycidyl ether , ß - estradiol 3 -methyl ether , B - estradiol O Me OR? 6 - one, ß - estradiol 3 -glycidyl , B - estradiol 6 - one 6 - ( 0 - car . 11110 boxymethyloxime) , 16 - epiestriol, 17 - epiestriol , 2 -methoxy 40 estradiol, 4 -methoxy estradiol, estradiol 17 -phenylpropi onate , and 17ß -estradiol 2 -methyl ether, 17a - ethynylestra diol, megestrol acetate , and estriol. E. As used herein , “ testosterone and derivatives thereof” IIZ refers to compounds, either naturally occurring or synthe sized , of Formula F : wherein R2, R3, RS, R7, R8, Rº, and R10 are each indepen dently moieties that permit or promote agonist or antagonist Formula F 50 activity upon the binding of the compound of Formula C to Me ORO a nuclear hormone receptor ; and each dash represents an optional double bond . In some embodiments , the structure of Formula C is substituted with one or more substituents at one or more positions of the tetracyclic ring , such as , for 55 example , positions 1 , 2 , 4 , 5 , 6 , 7 , 8 , 11 , 12 , 14 , and 15 . Specific , nonlimiting examples of derivatives of cortisol and derivatives thereof include cortisol, cortisone acetate , beclo (R ! O -- metasone , prednisone , prednisolone , methylprednisolone , betamethasone , trimcinolone , and dexamethasone. wherein R ' , when present, R2, R3 and Rº are each indepen - 60 As used herein , " linking group ” is a molecule or group of dently a moiety that permits or promotes agonist or antago - molecules that binds two separate entities to one another. nist activity upon binding of the compound of Formula F to Linking groups may provide for optimal spacing of the two a nuclear hormone receptor, and each dashed line represents entities or may further supply a labile linkage that allows the an optional double bond , with the proviso that no more than two entities to be separated from each other . Labile linkages one of the optional carbon - carbon double bond is present at 65 include hydrolyzable groups , photocleavable groups , acid position 5 . In some embodiments , the structure of Formula labile moieties , base -labile moieties and enzyme cleavable F is substituted with one or more substituents at one or more groups. US 9 , 783 ,592 B2 19 20 As used herein , the term “ prodrug” is defined as any 55 % , 60 % , 65 % , 70 % , 75 % , 80 % , 85 % , 90 % , or 95 % compound that undergoes chemical modification before identical to the corresponding sequence of native glucagon , exhibiting its full pharmacological effects . native oxyntomodulin , native exendin - 4 , native ( 7 -37 )GLP As used herein , a " dipeptide ” is the result of the linkage 1 , native GLP - 2 , or native GIP over the length of the native of an a - amino acid or a - hydroxyl acid to another amino 5 peptide ( or over the positions which correspond to glucagon , acid , through a peptide bond . see e .g ., FIG . 1 ). In other embodiments , a glucagon super As used herein the term “ chemical cleavage” absent any family peptide ( Q ) comprises an amino acid sequence of further designation encompasses a non - enzymatic reaction native glucagon , native exendin - 4 , native ( 7 -37 )GLP - 1 , that results in the breakage of a covalent chemical bond native GLP - 2 , native GHRH , native VIP, native PACAP - 27 , Embodiments 10 native PHM , native oxyntomodulin , native secretin , or The present disclosures provide glucagon superfamily native GIP with up to 1 , 2 , 3 , 4 , 5 , 6 , 7 , 8 , 9 or 10 amino acid peptides conjugated with NHR ligands. In some aspects, the modifications . In still further embodiments , Q comprises an NHR ligands are capable of acting at nuclear hormone amino acid sequence which is a chimera of two or more receptors involved in metabolism or glucose homeostasis , native glucagon related peptide sequences. In some embodi and the conjugate provides superior biological effects on 15 ments , Q comprises an amino acid sequence at least about metabolism or glucose homeostasis compared to the peptide 50 % identical to native glucagon (SEQ ID NO : 1601 ) that alone or the NHR ligand alone . Without being bound by a retains the alpha -helix conformation of the amino acids theory of the invention , the NHR ligand may serve to target corresponding to amino acids 12 - 29 . the glucagon superfamily peptide to particular types of cells In related aspects , the invention provides peptide conju or tissues ; or alternatively the glucagon superfamily peptide 20 gates represented by the formula may serve to target the NHR ligand or enhance its transport into the cell, e . g . through binding of peptide to a receptor Q - L - Y that internalizes the conjugate . wherein Q is osteocalcin , calcitonin , amylin , or an analog , The glucagon superfamily peptide conjugates of the derivative or conjugate thereof, rather than a glucagon invention can be represented by the following formula : 25 superfamily peptide ; Y is a NHR ligand ; and L is a linking group or a bond . In some embodiments , comprises Q - L - Y osteocalcin (SEQ ID NO : 1644 ) , or an amino acid sequence wherein Q is a glucagon superfamily peptide, Y is a NHR that is at least about 40 % , 45 % , 50 % , 55 % , 60 % , 65 % , 70 % , ligand , and L is a linking group or a bond . 75 % , 80 % , 85 % , 90 % , or 95 % identical to native osteocalcin The glucagon superfamily peptide ( Q ) in some embodi - 30 over the length of the native peptide . Q may comprise an ments can be a glucagon related peptide that exhibits agonist analog of osteocalcin with up to 1 , 2 , 3 , 4 , 5 , 6 , 7 , 8 , 9 or 10 activity at the glucagon receptor , agonist activity at the amino acid modifications relative to native osteocalcin , or a GLP - 1 receptor , agonist activity at the GIP receptor, co - truncated analog of osteocalcin ( e . g . , amino acids 70 - 84 ) agonist activity at the glucagon and GLP- 1 receptors , co with up to 1 , 2 , 3 , 4 , 5 , 6 , 7 , 8 , 9 or 10 amino acid agonist activity at the glucagon and GIP receptors , co - 35 modifications relative to the native truncated osteocalcin . In agonist activity at the GLP - 1 and GIP receptors , or tri - some embodiments , Q comprises calcitonin (SEQ ID NO : agonist activity at the glucagon , GIP , and GLP - 1 receptors. 1645 ) , or an amino acid sequence that is at least about 40 % , In some embodiments , the glucagon related peptide exhibits 45 % , 50 % , 55 % , 60 % , 65 % , 70 % , 75 % , 80 % , 85 % , 90 % , or antagonist activity at the glucagon , GLP - 1 or GIP receptor. 95 % identical to native calcitonin over the length of the The glucagon superfamily peptide ( Q ) in some embodi- 40 native peptide . Q may comprise an analog of calcitonin with ments may be a glucagon -related peptide , Growth Hormone up to 1 , 2 , 3 , 4 , 5 , 6 , 7 , 8 , 9 or 10 amino acid modifications Releasing Hormone (GHRH ; SEQ ID NO : 1619 ) , vasoac - relative to native calcitonin . In some embodiments , Q com tive intestinal peptide (VIP ; SEQ ID NO : 1620 ), Pituitary prises amylin (SEQ ID NO : 1646 ), or an amino acid adenylate cyclase - activating polypeptide 27 (PACAP - 27 ; sequence that is at least about 40 % , 45 % , 50 % , 55 % , 60 % , SEQ ID NO : 1621 ) , peptide histidine methionine (PHM ; 45 65 % , 70 % , 75 % , 80 % , 85 % , 90 % , or 95 % identical to native SEQ ID NO : 1622 ), or Secretin (SEQ ID NO : 1623 ) , and / or amylin over the length of the native peptide. Q may com and analogs, derivatives and conjugates thereof. Glucagon prise an analog of amylin with up to 1 , 2 , 3 , 4 , 5 , 6 , 7 , 8 , 9 superfamily peptides may have common structural charac - or 10 amino acid modifications relative to native amylin . teristics, including but not limited to homology within the The NHR Ligand ( Y ) N - terminal amino acids and / or alpha -helical structure within 50 In the present disclosures relating to Q - L - Y conjugates, Y the C -terminal portion . It is believed that the C - terminus is a ligand that acts at any nuclear hormone receptor, generally functions in receptor binding and the N - terminus including any one of the " nuclear hormone receptor super generally functions in receptor signaling . A few amino acids family ” (NHR superfamily ) set forth in Table 1 , or a class or in the N - terminal portion and C - terminal portion are highly subgroup thereof. This NHR superfamily is composed of conserved among members of the glucagon superfamily, for 55 structurally related proteins found within the interior of cells example , Hisl, Gly4 , Phe , Phe22, Val23 , Trp25, and that regulate the transcription of genes. These proteins Leu26 , with amino acids at these positions showing identity , include receptors for steroid and thyroid hormones, vita conservative substitutions or similarity in amino acid side mins, and other “ orphan ” proteins for which no ligands have chains. In some embodiments the glucagon related peptide been found . Nuclear hormone receptors generally include at Q is glucagon (SEQ ID NO : 1601) , oxyntomodulin ( SEQ ID 60 least one of a C4 -type zinc finger DNA - Binding Domain NO : 1606 ) , exendin - 4 (SEQ ID NO : 1618 ) , Glucagon - like (DBD ) and / or a Ligand Binding Domain (LBD ) . The DBD peptide - 1 (GLP - 1 ) (amino acids 7 - 37 provided as SEQ ID functions to bind DNA in the vicinity of target genes , and the NOs : 1603 and 1604 ), Glucagon - like peptide - 2 (GLP - 2 ) LBD binds and responds to its cognate hormone. “ Classical (SEQ ID NO : 1608 ) , GIP (SEQ ID NO : 1607 ) or analogs, Nuclear Hormone Receptors” possess both a DBD and a derivatives and conjugates of the foregoing . In some 65 LBD (e . g . Estrogen receptor alpha ), while other nuclear embodiments Q as a glucagon related peptide comprises an hormone receptors possess only a DBD ( e . g . Knirps, ORD ) amino acid sequence that is at least about 40 % , 45 % , 50 % , or only a LBD (e .g . Short Heterodimer Partner (SHP )) . US 9 , 783 ,592 B2 21 Nuclear hormone receptors can be divided into four proteins . Ligand binding to the Type II receptor causes mechanistic classes: Type I , Type II , Type III, and Type IV . dissociation of the corepressor and recruitment of coactiva Ligand binding to Type I receptors (NR3 Group ) results in tor proteins. Additional proteins are recruited to the nuclear the dissociation of heat shock proteins (HSP ) from the receptor/ DNA complex , which transcribe DNA into mes receptor, homodimerization of the receptor, translocation 5 senger RNA . Type III nuclear hormone receptors (NR2 from the cytoplasm into the cell nucleus , and binding to Group ) are orphan receptors that bind to direct repeat HRE ' s inverted repeat hormone response elements (HRE ' s ) of of DNA as homodimers . Type IV nuclear hormone receptors DNA . The nuclear receptor/ DNA complex then recruits bind to DNA either as monomers or dimers . Type IV other proteins which transcribe DNA downstream from the receptors are unique because a single DNA binding domain HRE into messenger RNA. Type II receptors (NR1 Group ) 10 of the receptor binds to a single half site HRE . The NHR are retained in the nucleus and bind as heterodimers , usually ligand can be a ligand that acts at any one or more of the with Retinoid X Receptors (RXR ), to DNA . Type II nuclear Type I, Type II, Type III or Type IV nuclear hormone hormone receptors are often complexed with corepressor receptors (e . g. as an agonist or antagonist ) . TABLE 1 Nuclear Hormone Receptor Superfamily Endogenous Nuclear Hormone Receptor Species Accession Ligand NR1 Group NR1A1 Thyroid Hormone Receptor alpha ( TRa ) Human M24748 Thyroid hormone NR1A2 Thyroid Hormone Receptor beta ( TRB ) Human X04707 NR1B1 Retinoic Acid Receptor alpha (RARa ) Human X06538 Vitamin A and NR1B2 Retinoic Acid Receptor beta (RARB ) Human Y00291 related compounds NR1B3 Retinoic Acid Receptor gamma (RARY ) Human M57707 NR1C1 Peroxisome Proliferator Activated Receptor alpha Human L02932 Fatty acids , ( PPARA ) prostaglandins NR1C2 Peroxisome Proliferator Activated Receptor Human L07592 beta /delta (PPARB / S ) NR1C3 Peroxisome Proliferator Activated Receptor gamma Human L40904 ( PPARY) NR1D1 Rev -Erbaa Human M24898 Heme NR1D2 Rev - ErbAB Human L31785 NR1F1 RAR - related Orphan Receptor alpha (RORA ) Human U04897 Cholesterol , all NR1F2 RAR - related Orphan Receptor beta (RORB ) Human Y08639 trans retinoic acid NR1F3 RAR -related Orphan Receptor gamma ( RORY) Human U16997 NR1H2 Liver X Receptor beta ( LXRB ) Human U07132 Oxysterol NR1H3 Liver X Receptor alpha ( LXRa ) Human U22622 NR1H4 Farnesoid X Receptor (FXR ) Human U68233 NR111 Vitamin D Receptor (VDR ) Human J03258 Vitamin D NR112 Pregnane X Receptor (PXR ) Human AF061056 Xenobiotics ( dexamethasone , rifapicin ) NR113 Constitutive Androstane Receptor alpha (CAR ) Human 230425 Androstane NR2 Group NR2A1 Hepatocyte Nuclear Factor 4 alpha (HNF4a ) TIITITLITTTTTTTTTTTTTTTTTTTTTTTTTTTTTTIHuman X76930 Fatty acids NR2A3 Hepatocyte Nuclear Factor 4 gamma (HNF4y ) Human Z49826 Fatty acids NR2B1 Retinoid X Receptor alpha (RXRa ) Human X52773 Retinoids NR2B2 Retinoid X Receptor beta (RXRB ) Human M84820 NR2B3 Retinoid X Receptor gamma (RXRY ) Human U38480 NR2C1 Testicular Receptor 2 ( TR2) Human M29960 NR2C2 Testicular Receptor 4 ( TR4 ) Human L27586 NR2E1 Human homologue of the Drosophila tailless gene Human Y13276 ( TLX ) NR2E3 Photoreceptor- specific Nuclear Receptor (PNR ) Human AF121129 NR2F1 Chicken ovalbumin upstream promoter Human X12795 transcription factor I ( COUP - TFI) NR2F2 Chicken ovalbumin upstream promoter Human M64497 transcription factor II (COUP - TFII ) NR2F6 V - erAA -Related Gene (EAR2 ) Human X12794 NR3 Group NR3A1 Estrogen Receptor alpha ( ERa ) Human P03372 Estrogen NR3A2 Estrogen Receptor beta (ERB ) Human AB006590 NR3B1 Estrogen Receptor Related alpha (ERRA ) Human X51416 NR3B2 Estrogen Receptor Related beta (ERRB ) Human AF094517 NR3B3 Estrogen Receptor Related gamma (ERRY ) Human AF058291 NR3C1 Glucocorticoid Receptor (GR ) Human X03225 Cortisol NR3C2 Mineralocorticoid Receptor (MR ) Human M16801 Aldosterone NR3C3 Progesterone Receptor (PR ) Human M15716 Progesterone NR3C4 Androgen Receptor (AR ) Human M20132 Testosterone US 9 ,783 ,592 B2 23 24 TABLE 1 - continued Nuclear Hormone Receptor Superfamily Endogenous Nuclear Hormone Receptor Species Accession Ligand NR4 Group NR4A1 Nerve Growth Factor IB alpha (NGFI - Ba ) Human L13740 NR4A2 Nerve Growth Factor IB beta (NGFI - BB ) Human X75918 NR4A3 Nerve Growth Factor IB gamma ( NGFI- BY) Human D78579 NR5 Group NR5A1 Steroidogenic Factor 1 (SF1 ) Human U76388 NR5A2 Liver Receptor Homolog- 1 (LRH - 1) Human U93553 NR6 Group NR6A1 Germ Cell Nuclear Factor (GCNF ) Human U64876 NROB Subgroup (have only LBD , no DBD ) NROB1 DAX1 Human S74720 NROB2 Short Heterodimer Partner (SHP ) Human L76571 Table data is taken from Laudet and Gronemeyer " The Nuclear Receptor Facts Book , " Academic Press . Class D refers to a classification code for each member, and accession refers to the NCBIGenBank nucleotide accession code . Activity of the NHR Ligand (Y ) ( 1997 ); Liu et al. , J. Biol. Chem ., 274 : 26654 -26660 (1999 ). In some embodiments , Y exhibits an EC50 for nuclear Binding of the NHR ligand to the nuclear hormone receptor hormone receptor activation (or in the case of an antagonist , 25 can be determined using any binding assay known in the art an IC50 ) of about 10 mM or less , or 1mM ( 1000 uM ) or less such as, for example , fluorescence polarization or a radio ( e . g ., about 750 uM or less , about 500 uM or less , about 250 active assay . See, e . g ., Ranamoorthy et al . , 138( 4 ): 1520 uM or less , about 100 uM or less, about 75 uM or less , about 1527 ( 1997 ) . 50 uM or less , about 25 uM or less , about 10 uM or less , In some embodiments, Y exhibits about 0 .001 % or more , about 7 . 5 uM or less, about 6 uM or less , about 5 uM or less , about 0 .01 % or more , about 0 . 1 % or more , about 0 . 5 % or about 4 uM or less , about 3 uM or less , about 2 uM or less more , about 1 % or more , about 5 % or more , about 10 % or or about 1 uM or less ). In some embodiments , Y exhibits an more, about 20 % or more , about 30 % or more , about 40 % EC50 or IC50 at a nuclear hormone receptor of about 1000 or more , about 50 % or more , about 60 % or more , about 75 % nM or less ( e . g . , about 750 nM or less , about 500 nM or less, 35 or more, about 100 % or more , about 125 % or more , about about 250 nM or less , about 100 nM or less , about 75 nM or 150 % or more , about 175 % or more , about 200 % or more , less , about 50 nM or less, about 25 nM or less , about 10 nM about 250 % or more , about 300 % or more , about 350 % or or less , about 7 . 5 nM or less , about 6 nM or less , about 5 nM more , about 400 % or more , about 450 % or more , or about or less , about 4 nM or less , about 3 nM or less , about 2 nM 500 % or higher activity at the nuclear hormone receptor or less or about 1 nM or less ) . In some embodiments , Y has 40 relative to the native nuclear hormone ( nuclear hormone an EC50 or IC50 at a nuclear hormone receptor which is in the potency ) . In some embodiments, Y exhibits about 5000 % or picomolar range . Accordingly , in some embodiments , Y less or about 10 , 000 % or less activity at the nuclear hormone exhibits an EC 50 or IC50 at a nuclear hormone receptor of receptor relative to native nuclear hormone . The activity of about 1000 PM or less ( e . g ., about 750 PM or less , about 500 Y at a receptor relative to a native ligand of the receptor is PM or less , about 250 PM or less , about 100 PM or less , 45 calculated as the inverse ratio of EC5os for Y versus the about 75 PM or less , about 50 PM or less, about 25 PM or native ligand . In some embodiments , Y is the native ligand less , about 10 pM or less , about 7 .5 PM or less , about 6 PM of the receptor. or less , about 5 PM or less , about 4 PM or less, about 3 PM Structure of the NHR Ligand ( Y ) or less , about 2 PM or less or about 1 PM or less ) . The NHR ligand of the invention ( Y ) is partly or wholly In some embodiments, Y exhibits an EC 50 or IC50 at a 50 non - peptidic and is hydrophobic or lipophilic . In some nuclear hormone receptor that is about 0 . 001 PM or more , embodiments , the NHR ligand has a molecular weight that about 0 .01 PM or more, or about 0 . 1 PM or more . Nuclear is about 5000 daltons or less , or about 4000 daltons or less , hormone receptor activation ( nuclear hormone receptor or about 3000 daltons or less , or about 2000 daltons or less , activity ) can be measured in vitro by any assay known in the or about 1750 daltons or less , or about 1500 daltons or less , art . For example , the activity at the nuclear hormone recep - 55 or about 1250 daltons or less , or about 1000 daltons or less , tor can be measured by expressing the receptor in yeast cells or about 750 daltons or less, or about 500 daltons or less , or also harboring a reporter gene ( e. g ., lacZ which encodes about 250 daltons or less. The structure of Y can be in B - galactosidase ) under the control of a hormone - responsive accordance with any of the teachings disclosed herein . promoter. Thus , in the presence of a ligand that acts at the In the embodiments described herein , Y is conjugated to receptor , the reporter gene is expressed and the activity of 60 L ( e . g . when L is a linking group ) or Q ( e . g . when L is a the reporter gene product can be measured ( e . g . , by mea - bond ) at any position of Y that is capable of reacting with Q suring the activity of B -galactosidase in breaking down a or L . One skilled in the art could readily determine the chromogenic substrate , such as chlorophenol red - ß - D - ga - position and means of conjugation in view of general lactopyranoside (CPRG ) , which is initially yellow , into a red knowledge and the disclosure provided herein . product that can be measured by absorbance ) . See , e . g ., 65 In any of the embodiments described herein wherein Y Jungbauer and Beck , J. Chromatog . B , 77 : 167 - 178 ( 2002 ) ; comprises a tetracyclic skeleton having three 6 -membered Routledge and Sumpter, J. Biol. Chem , 272 : 3280 -3288 rings joined to one 5- membered ring or a variation thereof US 9 , 783 ,592 B2 25 26 ( e . g . a Y that acts at the vitamin D receptor ), the carbon Contemplated optional substituents include, but are not atoms of the skeleton are referred to by position number, as limited to , OH , NH2, ketone, and C2 -C18 alkyl groups . shown below : In some embodiments , Y comprises a structure of For mula A wherein 5 R is present and is hydrogen , C1- C18 alkyl, C2- C18 alkenyl, C2- C18 alkynyl, heteroalkyl , (Co - C , alkyl) aryl, (Co Cg alkyl) heteroaryl , (Co - Cz alkyl) C (O ) C , - C18 alkyl, (C . -C . 18 20 24 alkyl) C (O )C2 - C18 alkenyl, (Co -C , alkyl )C (O )C2 - C18 alky . 26 nyl, (Co - C , alkyl) C ( O ) H , (Co - C , alkyl) C ( O ) aryl, (Co -Cg 19 11 25 10 alkyl) C ( O )heteroaryl , (Co - C , alkyl) C ( O )OC , -C18 alkyl , ( Co - C , alkyl) C ( O ) OC2- C18 alkenyl, (Co - Cg alkyl) C ( O ) OC2 N C18 alkynyl, (Co - C , alkyl) C ( O )OH , (Co - C , alkyl) C ( O ) O 10 aryl, (Co - C , alkyl) C ( O ) O heteroaryl , ( C . - C , alkyl) C ( O ) w NR24C , -C18 alkyl, ( C - C , alkyl) C ( O )NR24C2 -C18 alkenyl, 4 6 15 (Co -C , alkyl) C (O )NR24C2 - C18 alkynyl , (Co - C , alkyl) C (O ) NR24H2, (C . - C , alkyl) C ( O )NR24 aryl, (C .- C , alkyl) C (O ) NR24heteroaryl, or SOZH ; For example , a modification having a ketone at position -6 R2 is present and is hydrogen , (Co -Cg alkyl) halo , C7 -C18 refers to the following structure: alkyl , C2 -C18 alkenyl, C2- C18 alkynyl, heteroalkyl , (Co -C , 20 alkyl) aryl, (Co - C , alkyl) heteroaryl , (Co - C , alkyl) OC , -C18 alkyl , (Co - Cz alkyl) OC2 - C18 alkenyl , (Co - Cz alkyl )OC2 - C18 | 21 22 alkynyl, (Co -C , alkyl) OH , (C . - C , alkyl )SH , (C . - C , alkyl ) 18 24 NR24C , -C18 alkyl, (Co -C , alkyl )NR24C2 - C18 alkenyl, (Co 23 26 Cg alkyl )NR24C2 - C18 alkynyl, (Co - Cg alkyl )NR24H2 , (Co - C8 19 (11 13 25 alkyl) C ( O ) C , -C18 alkyl, ( Co- C , alky1) C ( O )C2 - C18 alkenyl , (Co -C , alkyl) C (O ) C2- C18 alkynyl , (Co - Cg alkyl )C (O ) H , (Co N al C , alkyl ) C ( O ) aryl, (Co - C , alkyl) C ( O ) heteroaryl, (Co - C , alkyl ) C ( O )OC -Cis alkyl, (Co - Cz alkyl ) C ( O )OCZ - C18 alk w enyl, (Co -Cg alkyl) C ( O )OC2 -C18 alkynyl, (Co - Cg alkyl) C ( O ) 30 OH , (Co - C , alkyl) C ( O ) O aryl , (Co - C , alkyl ) C ( O ) O het eroaryl, (Co - C , alkyl )OC ( O ) C , -C18 alkyl, (Co - C , alkyl) OC ( 0 )C2 - C18 alkenyl, (Co -C , alkyl) OC (O )C2 - C18 alkynyl, (Co NHR Ligand that Acts on a Type I Nuclear Hormone Cg alkyl) C ( O )NR24C7 -C18 alkyl, (C . - C , alkyl) C (O ) Receptor NR24C2- C18 alkenyl, (Co - Cg alkyl) C (O )NR24C2 - C18 In some embodiments of the invention , the NHR ligandº35 alkynyl, (Co - C , alkyl) C (O )NR24H2 , (Co - C , alkyl) C ( O ) ( Y ) acts on a Type I nuclear hormone receptor. In some NR24aryl , (C . -C , alkyl) C (O )NR24heteroaryl , (Co -C , alkyl) embodiments , Y can have any structure that permits or NR24C ( O ) C -C18 alkyl, ( C . - C alkyl) NR24 C ( O )C2 - C18 alk promotes agonist activity upon binding of the ligand to a enylIKVI , NRor 24COOH(Co - C , alkyl . C )NR24C . - C . alkylocIOOC( O ) C2- C18 alkynyl . - C , (Co alkyl - C ,. Type I nuclear hormone receptor, while in other embodi- 40 (Co - C , alkyl) OC ( O )OC , -Cie alkenyl, ( C . - C , alkyl) OC ( O ) ments Y is an antagonist of the- Type 1I nuclearnuclear hormone OC2- C18 alkynyl, (Co - C , alkyl) OC (O )OH , (Co - C , alkyl) OC receptor. ( O )NR24C7 - C18 alkyl, (Co - C , alkyl) OC ( O )NR24C2 - Cig alk In exemplary embodiments , Y comprises a structure as enyl , ( C . - C . alkyl) OC ( O )NR24C , - Cie alkynyl, ( C . - C . shown in Formula A : alkyl )OC ( O )NR24H2 , (Co - Cg alkyl )NR24 (O )OC , - C18 alkyl , 45 (Co -Cg alkyl) NR24 (O ) OC2- C18 alkenyl, (Co - C , alkyl)NR24 ( O )OC2 - C18 alkynyl , or (Co - C , alkyl) NR24 (O )OH ; Formula A R² is hydrogen , (Co - C , alkyl) halo , C , - C18 alkyl, C2- C18 18 alkenyl, C2- C18 alkynyl, heteroalkyl, (Co - Cg alkyl) aryl, (Co 12 Me R4 C , alkyl )heteroaryl , (Co - C , alkyl) OC , - C18 alkyl, ( C .- C , 196 50 alkyl) OC2 - C18 alkenyl, (Co - C8 alkyl) OCZ - C18 alkynyl, (Co (R2 ) . Cg alkyl) OH , (Co -C , alkyl )SH , (Co -C , alkyl )NR24C , - C18 alkyl, (Co - C , alkyl )NR24C2 - C18 alkenyl, (Co - C , alkyl) 18 NR24C2 -C18 alkynyl, (C . -C , alkyl) NR24H2 , (C . -C , alkyl) C IM ( O ) C7 -C18 alkyl, ( C . - C . alkyl ) C ( O )C2 -C18 alkenyl , ( Co -Cg (R ! --- - 55 alkyl) C (O ) C2- C18 alkynyl , (Co -C , alkyl) C ( O ) H , (Co - C , alkyl ) C ( O ) aryl, ( Co- C , alkyl ) C ( O )heteroaryl , (Co - C , alkyl) C ( O ) OC1- C18 alkyl, (Co - C , alkyl) C ( O )OC2 - C18 alkenyl , wherein Rand R ’ , when present, are independently moi- (Co - C , alkyl) C ( O ) OC2 -C18 alkynyl, (Co - C , alkyl) C ( O )OH , eties that permit or promote agonist or antagonist activity (Co - C , alkyl ) C ( O ) O aryl , (Co -Cg alkyl) C ( O ) O heteroaryl , upon binding of the compound of Formula A to the Type I 60 (Co - C , alkyl) OC (O ) C -C18 alkyl, (Co - C , alkyl )OC (O )C2 nuclear hormone receptor; R and R + are independently C18 alkenyl, (Co - C , alkyl) OC ( O ) C2 -C18 alkynyl, (Co - C , moieties that permit or promote agonist or antagonist activ - alkyl) C ( O )NR24C7 - C18 alkyl, (Co - C , alkyl ) C ( O )NR24C2 ity upon binding of the compound of Formula A to the Type C18 alkenyl, (Co - C , alkyl) C ( O ) NR24C2- C18 alkynyl, (Co - C , I nuclear hormone receptor , and each dashed line represents alkyl) C (O )NR24H2 , (Co -C , alkyl) C ( O )NR24aryl , (Co -C 00 an optional double bond . Formula A may further comprise 65 alkyl) C (O )NR24heteroaryl , (C . -C , alkyl) NR24C ( O )C7 - C18 one or more substituents at one or more of positions 1 , 2 , 3 , alkyl, (Co -C , alkyl) NR24C ( O )C2 - C18 alkenyl, or ( Co- C , 4 , 5 , 6 , 7 , 8 , 9 , 11 , 12 , 14 , 15 , 16 , 17 , 18 , and 19 . alkyl) NR24C ( 0 )C2 - C18 alkynyl, ( C . - C , alkyl) NR24C ( O ) US 9 ,783 , 592 B2 27 28 OH , (C .- C , alkyl )OC (O )OCZ - C18 alkyl , (Co -C , alkyl )OC OC (O )NR24C7 -C , alkyl, ( C .- C , alkyl) OC ( O )NR24C2 - C3 (O )OC2 -C18 alkenyl, (Co - C , alkyl) OC ( O )OC2 - C18 alkynyl, alkenyl , (C .- C , alkyl) OC ( O )NR24C2 - C , alkynyl , ( C . - C . (Co - C , alkyl )OC ( O )OH , (Co - C , alkyl )OC ( O )NR24C7 - C18 alkyl) OC ( O )NR24H2 , (Co - C , alkyl) NR24 ( O )OC , - C , alkyl, alkyl, (Co - C , alkyl) OC ( O ) NR24C2- C18 alkenyl, (Co - Co (Co - C , alkyl) NR24 ( O ) OC2- C , alkenyl, (Co - C , alkyl) NR24 alkyl) OC ( O )NR24C2 - C18 alkynyl, (Co -Co alkyl) OC ( O ) 5 ( O ) OC . - C , alkynyl, or ( C . - C , alkyl) NR24 ( 0 )OH ; and , NR24H ,, (Co - Cg alkyl) NR24 (O )OC , -C18 alkyl, (Co -Cg alkyl) R24 is hydrogen or C , -C , alkyl. NRC ( O )OC2 - C18 alkenyl, (Co - Cg alkyl) NRC ( O )OC2 - C18 In some embodiments , R1 is hydrogen , propionate , alkynyl, or (Co - C , alkyl) NR24 ( O ) OH ; R4 is hydrogen , (Co - C , alkyl) halo , C7 -C18 alkyl, C2 -C18 acetate , benzoate , or sulfate ; R2 is hydrogen or methyl; R * is alkenyl, C2 -C18 alkynyl , heteroalkyl, (Co - C , alkyl) aryl, (Co - 10 hydrogen or methyl; and R * is acetate , cypionate , hemisuc C , alkyl )heteroaryl , (Co - C , alkyl) OC -C18 alkyl, ( C . - C ciniate , enanthate , or propionate . alkyl) OC2 - C1g alkenyl, (Co - Cg alkyl) OC2 - C18 alkynyl, (Co In embodiments wherein Y comprises a structure of Ce alkyl )OH , (Co - C , alkyl ,SH , (Co - Co alkyl) NR24C7 - C180 Formula A , Y is conjugated to L ( e . g . when L is a linking alkyl , (Co - C , alkyl) NR24C2 - C18 alkenyl , ( Co- C , alkyl) group ) or Q ( e. g . when L is a bond ) at any position of NR24C . - C . . alkynyl, ( C . - C . alky1NR24H . . ( C . - C . alkvi? 15 Formula A that is capable of reacting with Q or L . One ( 0 )C , -C , alkyl, (C . - C , alkyl )C (O ) C2- C18 alkenyl, ( C .- C . skilled in the art could readily determine the position of alkyl) C ( O )C2 - C18 alkynyl, (Co - C , alkyl ) C (O )H , (Co - C conjugation on Formula A and means of conjugation of alkyl) C ( O ) aryl, ( C . - C , alkyl ) C ( O )heteroaryl , (Co - C , alkyl) Formula A to Q or L in view of general knowledge and the C ( O ) OC -C18 alkyl , (Co - C , alkyl) C ( O )OC2 - C18 alkenyl, disclosure provided herein . In some embodiments , Formula (Co - C , alkyl) C ( O ) OC , -Cig alkynyl, (Co - C , alkyl) C ( O )OH , 20 A is conjugated to L or Q at any of positions 1 , 2 , 3 , 4 , 5 , 6 , ( Co- C , alkyl) C (O )O aryl, ( C .- C , alkyl) C ( O )O heteroaryl , 7 , 8 , 9 , 10 , 11 , 12 , 13 , 14 , 15 , 16 , 17 , 18 , 19 , or 20 of Formula (Co - C , alkyl) OC (O )C . -C , alkyl, ( C . -C , alkyl) OC (O )C2 - C , A . In some embodiments , Formula A is conjugated to L or alkenyl, (C . -C , alkyl) OC (O )C , - C , alkynyl, (Co - C , alkyl) Q at position 1 , 3 , 6 , 7 , 12 , 10 , 13, 16 , 17 , or 19 of Formula C ( O )NR24C , -C18 alkyl , (Co -Cg alkyl) C ( O )NR24C2 - C18 alk - A . enyl , ( C . - C , alkyl) C ( O )NR24C . - C , alkynyl, (Co - C , alkyl ) 25 In some embodiments , Y acts at an estrogen receptor ( e . g . C ( O )NR24H , ( C . - C . alkyl ) C ( O )NR24 aryl, ( C . - C . alkyl c ERA , ERB ) . In some embodiments , Y permits or promotes ( O )NR24heteroaryl , ( Co - C , alkyl) NR24C ( O ) C ; -C18 alkyl, agonist activity at the estrogen receptor, while in other ( C -C , alkyl) NR24C (O ) C2- C18 alkenyl , or (Co - Cz alkyl) embodiments Y is an antagonist of ER . In exemplary NR24C (O )C2 - C18 alkynyl , (Co -Cz alkyl) NR24C (O )OH , (Co - embodiments , Y can have a structure of Formula B : Cg alkyl )OC ( O )OC , -C18 alkyl, (Co - C , alkyl )OC ( O )OCZ - 30 Cig alkenyl, (Co -Cg alkyl) OC ( O )OC2 - C18 alkynyl, (Co - Co alkyl) OC ( O )OH , ( Co- C , alkyl) OCONR24C7 - C18 alkyl, Formula B (C . -C , alkyl) OC (O )NR24C2 - C18 alkenyl, ( C .- C , alkyl )OC Me ORO (CoO ) NR24C2 alkyl) NR24 - C18 ( O alkynyl ) OC -C8, (Co -alkylCg alkyl, (Co ) OC - C ,(O )alkylNR24H2 ) NR24 , (Co ( 0 ) 35 OURS OC2- C18 alkenyl, (Co -Cg alkyl )NR24 (O )OC2 - C18 alkynyl, or (C .- C alkyl) NR24 ( O )OH ; and R24 is hydrogen or C , -C18 alkyl. In some embodiments , Y comprises a structure of For mula A wherein 40 R ' is present and is hydrogen , C7 - C , alkyl; (Co -Cz alkyl) C ( O ) C , - C , alkyl, (Co -Cz alkyl) C ( O ) aryl, or SOzH ; wherein R ', RS and R? are moieties that permit or promote R is present and is hydrogen , halo , OH , or C , -C , alkyl; agonist or antagonist activity upon binding of the compound R® is hydrogen , halo , OH , or C - C , alkyl; of Formula B to the estrogen receptor. In some embodi R4 is hydrogen , (Co - C , alkyl )halo , C , -C , alkyl, C2- C , 45 ments , Formula B further comprises one or more substi alkenyl , C2- 18 alkynyl, heteroalkyl , ( Co- C , alkyl ) aryl , (Co - tutents at one or more of positions 1 , 2 , 4 , 6 , 7 , 8 , 9 , 11, 12, C , alkyl) heteroaryl , (Co - C , alkyl) OC , - C , alkyl, ( Co- C , 14 , 15 , and 16 ( e . g . a ketone at position - 6 ) . alkyl) OC2 -Cg alkenyl, (Co - Cg alkyl) OC2 -Cg alkynyl, (Co -C8 In some embodiments when Y comprises a structure of alkyl) OH , (C .- C alkyl) SH , (C .- C , alkyl) NR24C ,- C , alkyl, Formula B , wherein ( Co - C , alkyl) NR24C2 - Cg alkenyl, ( Co - C , alkyl) NR24C2 - Cg 50 R is hydrogen , C . - C18 alkyl, C2- C18 alkenyl, C2- C18 alkynyl, (Co - C , alkyl) NR24H2 , (Co - C , alkyl) C ( O ) C7 - C alkynyl, heteroalkyl, (Co - C , alkyl) aryl, (Co - Cg alkyl )het alkyl , (Co - C , alkyl ) C ( O )C2 - C , alkenyl, (Co - C , alkyl) C ( 0 ) eroaryl , (Co - C , alkyl) C ( O )C7 -C18 alkyl, (Co - C , alkyl ) C ( O ) C2- C , alkynyl, ( C . - C . alkyl ) C ( O ) H , ( C . - C . alkyl) C ( O ) aryl, C2 -C18 alkenyl , (Co - Cg alkyl) C ( O )C2 - C18 alkynyl, ( Co- Cg ( C . - C . alkyl) C ( O )heteroaryl , (Co - C , alkyl) C ( O ) OC , - C , alkyl) C ( O ) H , ( C . - C . alkyl) C ( O ) aryl , (Co - C , alkyl) C ( O )het alkyl, (Co - Cg alkyl) C ( O )OC2 - Cg alkenyl, (Co - Cg alkyl) C ( O ) 55 eroaryl, (Co - C , alkyl) C ( O )OC , -C18 alkyl, (Co - C , alkyl) C OC2- C , alkynyl, (Co - C , alkyl )C (O )OH , (Co -C , alkyl) C (O )OC2 - C18 alkenyl, (Co - C , alkyl ) C ( O ) OC2- C18 alkynyl, ( 0 ) 0 aryl, (Co - C , alkyl) C ( O ) O heteroaryl, (Co - C , alkyl) OC (Co - C , alkyl) C ( O )OH , C . - C , alkyl) C ( O ) O aryl, (Co - C , (0 )C ,- C , alkyl, (Co - Cg alkyl) OC ( O )C2 - Cg alkenyl, (Co - C , alkyl) C (O ) O heteroaryl, (Co - C , alkyl) C ( O )NR24C , -C18 alkyl) OC ( O )C2 - C18 alkynyl, (Co - C , alkyl ) C ( O )NR24C7 - C , alkyl, (Co - C , alkyl ) C ( O )NR24C2 - C18 alkenyl, (Co - C , alkyl) alkyl, (Co - Cg alkyl ) C ( O )NR24C2 - C , alkenyl, ( Co- C , alkyl ) 60 C ( O )NR24C2 - C18 alkynyl, (Co - C , alkyl) C ( O ) NR24H2, (Co C (O )NR24C2 - C , alkynyl, (Co -Cz alkyl) C ( O )NR24H ,, (Co - C , alkyl) C ( O )NR24aryl , (Co -Cz alkyl) C (O )NR24heteroaryl , Cg alkyl) C (O ) NR24aryl, (Co - C , alkyl) C (O )NR24heteroaryl , or SOZH ; (Co - C , alkyl) NR24C ( O ) C , - C , alkyl, (Co - C , alkyl) NR24C R $ is hydrogen , (Co -Cg alkyl) halo , C7 -C18 alkyl , C2 -C18 ( 0 )C2 - Cg alkenyl, or (Co -Cg alkyl) NR24C ( O ) C2- Cg alkynyl, alkenyl, C2- C18 alkynyl, heteroalkyl, (Co - C , alkyl) aryl, (Co (Co - Cg alkyl )NR24C ( O )OH , (Co - Cg alkyl) OC ( O )OC , -Cg 65 C , alkyl) heteroaryl , (Co - C , alkyl )OCZ -C18 alkyl , (Co - Cg alkyl, (Co - Cg alkyl )OC (O )OC2 - Cg alkenyl , (C . -C , alkyl )OC alkyl )OC2 - C18 alkenyl, (C .- C alkyl) OC2- C18 alkynyl , (Co ( O )OC2 - C , alkynyl, (Co - C , alkyl )OC ( O )OH , (Co - C , alkyl ) Cg alkyl) OH , (Co - C , alkyl )SH , (Co - Cg alkyl) NR24C2 - C18 US 9 ,783 , 592 B2 29 30 alkyl, (Co - Cz alkyl) NR24C2 - C18 alkenyl, ( Co- C , alkyl ) alkyl) OC ( O )C2 - C18 alkynyl, (Co - C , alkyl ) C (O ) NR240 , - C , NR24C2- C18 alkynyl, (Co -Cg alkyl )NR24H2 , (Co - C , alkyl) C alkyl , (C . -C alkyl) C ( O ) NR24C2- C , alkenyl , (Co - C , alkyl) (0 )C ,- C , alkyl, (Co - C , alkyl) C ( O )C2 - C18 alkenyl, (Co -C , C (O )NR24C2 -Cg alkynyl, (Co -C , alkyl) C ( O )NR24H2 , (Co alkyl) C ( O )C2 - C18 alkynyl, (Co - C , alkyl) C ( O ) H , ( C . - C Cg alky1 ) C ( O )NR24 aryl, ( Co- C , alkyl) C ( O )NR24heteroaryl , alkyl) C ( O ) aryl, (Co - C , alkyl) C ( O )heteroaryl , (Co - Cg alkyl) 5 (Co - C , alkyl) NR24C ( O ) C . - C , alkyl , (Co - C , alkyl) NR24C C ( O )OC , -C18 alkyl, (Co - C , alkyl) C ( O ) OC2- C18 alkenyl, (0 ) C2- Cg alkenyl , or (Co - Cg alkyl )NR24C ( O )C2 - C , alkynyl , (C . -C , alkyl) C ( O )OC2 - C18 alkynyl , (Co - C , alkyl) C (O )OH , ( Co- C , alkyl) C ( O ) O aryl, (Co - C , alkyl) C ( O ) O heteroaryl, (Co - C , alkyl )NR24C ( O )OH , (Co - C , alkyl) OC ( O ) OC , - C , ( Co- Cz alkyl) OC ( O ) C , -Cg alkyl, ( C . - C , alkyl )OC ( O )C2 -C18 alkyl, (Co - Cg alkyl )OC ( O )OCZ - C , alkenyl, ( C - C alkyl) OC alkenyl, (Co - Cg alkyl) OC ( O )C2 - C18 alkynyl, (Co - C , alkyl ) 10 (O ) OC2- C , alkynyl, ( Co - C , alkyl )OC ( O )OH , ( Co- Cg alkyl) C (O )NR24C7 -C18 alkyl, (C .- C , alkyl) C (O )NR24C2 - C18 alk OC( O )NR24C , - C , alkyl , (Co - C , alkyl) OC (O )NR24C2 - Cg enyl, (Co -C , alkyl) C (O )NR24C2 -C18 alkynyl, (Co -C , alkyl) alkenyl, ( C . - C , alkyl) OC ( O )NR24C2 - C , alkynyl , ( C . -Cg C ( O )NR24H2 , (C .- C , alkyl) C ( O ) NR24aryl, (Co -C , alkyl) C alkyl) OC (O )NR24H2 , ( C .- C , alkyl )NR24 (0 )0C ; -C , alkyl, ( O )NR24heteroaryl , (Co - C , alkyl) NR24C ( O ) C , -C18 alkylikui, (Co - C , alkyl )NR24 ( O )OC2 - C , alkenyl , (Co - C , alkyl) NR24 (Co -C , alkyl) NR24C ( O )C2 - C18 alkenyl, or (Co - C , alkyl ) 15 ( 02 g alkynyt, or ( ogalky )NE TOOH ; NR24C ( O )C2 -C18 alkynyl, ( C - C , alkyl) NR24C ( O ) OH , ( C . R® is hydrogen , C , -C alkyl, C2- C , alkenyl, C2- C , alky C , alkyl) OC ( O )OC , - Cig alkyl, (Co - C , alkyl) OC ( O )OC , - nyl , heteroalkyl, (Co - Cg alkyl) aryl , (Co -Cg alkyl) heteroaryl , Cig alkenyl, (Co -C , alkyl) OC (O )OC2 -C18 alkynyl, (Co - C (Co - Cg alkyl) C (O )C - Cg alkyl, ( Co -Cg alkyl) C ( O ) C2 - Cg alk alkyl) OC (O )OH , ( C . - C , alkyl) OC (O )NR240 , -C18 alkyl, enyl, ( Co - Cg alkyl) C (O )C2 - Cg alkynyl, (Co - Cg alkyl) C (O ) H , (Co - C , alkyl) OC ( O ) NR24C2- C18 alkenyl, (Co - C , alkyl) OC 20 (Co - C , alkyl) C ( O )aryl , (Co - Cg alkyl) C ( O )heteroaryl , ( C . - C , (O )NR24C2 - C18 alkynyl, (C .- C . alkyl) OC (O )NR24H2 , (Co alkyl) C (O )OC , -C , alkyl, (Co - C , alkyl) C (O )OC2 - Cg alkenyl, Cg alkyl )NR24 (O )OC -C18 alkyl, (Co - C , alkyl) NR24 (0 ) (Co - C , alkyl) C ( O ) OC2- Cg alkynyl, (Co - C , alkyl) C ( O )OH , OC2- Cig alkenyl, (Co -Cz alkyl )NR24 ( 0 )2C2 - C18 alkynyl, or Co - C , alkyl) C (O ) O aryl , (Co - C , alkyl ) C ( O )O heteroaryl, (Co - C , alkyl) NR24 (O )OH ; (Co - C , alkyl) C ( 0 ) C , -C18 alkyl, (Co -C , alkyl )C ( O ) NR24C2 - C , alkyl, ( C . - C , alkyl) C ( O ) (Co -C , alkyl) C (O )C2 - C18 alkenyl, (Co - Cg alkyl) C (0 )C2 -C18 25 NR24C2 - Cg alkenyl, (Co - C , alkyl) C (O )NR24C2 -C , alkynyl, alkynyl, (Co -C , alkyl) C (O )OC ,- C18 alkyl, ( Co- C , alkyl )C (Co - C , alkyl) C (O )NR24H2 , (Co - C , alkyl ) C ( O )NR24aryl , or ( O )OC2 - C18 alkenyl , (Co - C , alkyl) C ( O ) OC2- C18 alkynyl, (C .- C , alkyl )C (O )NR24heteroaryl ; and (Co - C , alkyl) C ( O ) OH , ( C . - C , alkyl) OC ( O ) C7 -C18 alkyl, R24 is hydrogen or C , - C , alkyl . (Co - Cg alkyl) OC (O )C2 - C18 alkenyl, (Co - Cg alkyl) OC (O )C2 - For example , R ' is hydrogen , propionate , acetate , benzo C18 alkynyl, (Co - C , alkyl) C ( O )NR24C7 -C18 alkyl, ( C . - C , 30 ate , or sulfate ; R is hydrogen , ethynyl, hydroxyl; and R? is alkyl) C (O )NR24C2 - C18 alkenyl, (Co - C , alkyl ) C ( O )NR24C2 acetate , cypionate , hemisucciniate , enanthate , or propionate . C18 alkynyl, (Co - C , alkyl) C ( O ) NR24H , (Co - C , alkyl) Nonlimiting examples of the compound of Formula B NR24C (O )C , - C18 alkyl, (Co - C , alkyl) NR24C (O )C - C , alk - include 17ß -estradiol , modified forms of estradiol such as enyl, or (Co - C , alkyl) NR24C (O )C2 - C18 alkynyl, or (Co - Cg B -estradiol 17 - acetate , ß -estradiol 17 -cypionate , ß -estradiol alkyl) NR24C ( O )OH ; 35 17 - enanthate , ß -estradiol 17 - valerate , 3 -estradiol 3 , 17 -diac Rº is hydrogen , C .- C18 alkyl, C2- C18 alkenyl, C2 -C18 etate , ß -estradiol 3 , 17 -dipropionate , ß -estradiol 3 -benzoate , alkynyl, heteroalkyl, (Co - C , alkyl) aryl , (Co -C , alkyl )het B - estradiol 3 - benzoate 17 - n -butyrate , B - estradiol 3 - glycidyl eroaryl , (Co - C , alkyl) C ( O )C7 -C18 alkyl, (Co - C , alkyl) C ( O ) ether , ß -estradiol 3 -methyl ether , B -estradiol 6 - one , B -estra C2- C18 alkenyl, (Co - C , alkyl) C ( O ) C2- C18 alkynyl, ( Co -C , diol 3 - glycidyl, ß -estradiol 6 -one 6 - ( 0 -carboxymethylox alkyl) C ( O ) H , (Co - C , alkyl) C (O )aryl , (Co -C , alkyl) C (O )het - 40 ime) , 16 - epiestriol, 17 - epiestriol, 2 -methoxy estradiol, eroaryl, (Co - C , alkyl) C (O )OCZ -C18 alkyl, (Co -C , alkyl) C 4 -methoxy estradiol , estradiol 17 - phenylpropionate , and ( O ) OC2- C18 alkenyl, (Co - C , alkyl) C ( O ) OC2- C18 alkynyl, 17ß -estradiol 2 -methyl ether , 17a -ethynylestradiol , mege (Co - C , alkyl) C ( O )OH , C . - C , alkyl) C ( O ) O aryl, ( C . - C . strol acetate , estriol, and derivatives thereof. In some alkyl) C ( 0 ) 0 heteroaryl, (Co - C , alkyl) C ( O )NR24C7 -C18 embodiments , carbon 17 has a ketone substitutent and RS alkyl, (C -C , alkyl) C ( O )NR24C2 - C18 alkenyl, (Co - C , alkyl) 45 and Rºare absent (e . g. estrone ) . Some of the aforementioned C ( O )NR24C2 -C18 alkynyl, (Co - C , alkyl) C ( O ) NR24H , (Co - compounds of Formula B are shown below : Cg alkyl) C ( O )NR24aryl , ( Co- C , alkyl) C ( O )NR24heteroaryl , or SOzH ; and , R24 is hydrogen or CZ - C18 alkyl. Me OH , In some embodiments , Y comprises a structure of For- 50 mula B , wherein Rl is hydrogen , C , -C , alkyl; (Co -Cz alkyl) C (0 ) C -C , alkyl, (Co - Cz alkyl) C ( O ) aryl, or SOzH ; R® is hydrogen , (Co - C , alkyl) halo , C , - C , alkyl, C2- C , alkenyl, C2- 18 alkynyl , heteroalkyl , (Co - C , alkyl ) aryl, (Co - 55 C , alkyl) heteroaryl , (Co -C , alkyl) OC - C alkyl, (Co - Cg HO alkyl) OC2 - Cg alkenyl, (Co - Cg alkyl) OC2 - C , alkynyl, (Co - C , Estradiol alkyl) OH , (Co - C , alkyl) SH , (Co - C , alkyl) NR24C , - C , alkyl, Me OH ( Co - C , alkyl) NR24C2 - Cg alkenyl, ( Co - C , alkyl) NR24C2 - Cg alkynyl, (Co - C , alkyl )NR24H , (Co - C , alkyl) C (O ) C - C , 60 alkyl, (Co - C , alkyl) C ( O )C2 -C , alkenyl, (Co -C , alkyl) C (O ) > 111111OH . C2- C , alkynyl , (Co - C , alkyl) C ( O ) H , (Co - C , alkyl) C ( O ) aryl, (Co - C , alkyl ) C ( O ) heteroaryl, (Co -C , alkyl) C (O )OC ,- C , alkyl, (Co -Cg alkyl) C ( O )OC2 -Cg alkenyl, (Co -Cg alkyl) C ( O ) OC2 - C , alkynyl, (Co - Cg alkyl) C (O )OH , ( Co - C , alkyl) C 65 HO ( 0 ) aryl, (Co - Ce alky1 ) C ( O ) O heteroaryl, ( C . - C . alkyl) OC Estriol (O ) C - C , alkyl, (Co - Cg alkyl )OC ( O )C2 - C , alkenyl, (Co - C8 Doment US 9 ,783 ,592 B2 31 32 - continued other embodiments Y is an antagonist of GR . In exemplary Me , and embodiments , Y comprises a structure of Formula C :

Formula C R6 27 III Me HO Estrone 10 Me OH --- - 1 = O= 15 ?H wherein R ? , R3, R , R7, RS, Rº, and R10 are each indepen dently moieties that permit or promote agonist or antagonist HO activity upon the binding of the compound of Formula C to Ethylyl estradiol the GR ; and each dash represents an optional double bond . 20 In some embodiments , Formula C further comprises one or In embodiments wherein Y comprises a structure of more substituents at one or more of positions 1 , 2 , 4 , 5 , 6 , Formula B , Y is conjugated to L ( e .g . when L is a linking 7 , 8 , 9 , 11, 12 , 14 , and 15 (e . g. hydroxyl or ketone at group ) or Q ( e . g . when L is a bond ) at any position of position - 11 ) . Formula B that is capable of reacting with Q or L . One In some embodiments , Y comprises a structure of For skilled in the art could readily determine the position of 25 mula C wherein conjugation on Formula B and means of conjugation of R² is hydrogen , (Co - Cg alkyl) halo , C , -C18 alkyl, C2 -C18 Formula B to Q or L in view of general knowledge and the alkenyl, C2 -C18 alkynyl, heteroalkyl, ( Co - Cg alkyl) aryl, (Co disclosure provided herein . In some embodiments , Formula Cg alkyl) heteroaryl , (Co - Cg alkyl) OC -C18 alkyl, (Co - Cg alkyl) OC2 - C18 alkenyl, (Co - C , alkyl) OC2 - C18 alkynyl , (Co B is conjugated to L or Q at any of positions 1 , 2 , 3 , 4 , 5 vla, 6 , 30 Cg alkyl) OH , (Co - C , alkyl) SH , (Co - Cg alkyl) NR24C1 - C18 7 , 8 , 9 , 10 , 11 , 12 , 13 , 14 , 15 , 16 , 17 , 18 , 19 , or 20 of Formula 30 alkyl , (Co - Cz alkyl )NR24C2 - C18 alkenyl, (Co - C , alkyl ) B . In some embodiments , Formula B is conjugated to L or NR24C2 -C18 alkynyl, (Co -C , alkyl )NR24H2 , (C . - C , alkyl) C Q at position 3 or 17 of Formula B . (O ) C , -C18 alkyl, (Co - C , alkyl ) C ( O )C2 - C , alkenyl, (Co - C , In other embodiments , Y acts at an estrogen receptor but alkyl ) C (O )C2 - C18 alkynyl , (Co - C , alkyl) C ( O ) H , ( Co -Cg is not encompassed by Formula B . Nonlimiting examples of 35 alkyl) C ( O ) aryl , (Co - C , alkyl) C ( O )heteroaryl , ( Co- C , alkyl) ligands that act at an estrogen receptor that are not encom C ( O ) OC , -C18 alkyl, (Co - C , alkyl ) C ( O )OC2 - C18 alkenyl, passed by Formula B are shown below : (Co - C , alkyl) C ( O )OC2 -C19 alkynyl, (Co - Cg alkyl) C ( O ) OH , (Co -C , alkyl) C (O )O aryl, (C .- C , alkyl) C ( O )O heteroaryl , (Co - C , alkyl) OC ( O ) C -C18 alkyl, (Co - C , alkyl )OC ( O )C2 40 Cis alkenyl, (Co - C , alkyl) OC (O )C2 - Cis alkynyl , (Co - Co H3CM alky1 ) C ( O )NR24C7 -Cig alkyl, (Co - C , alkyl) C ( O )NR24C2 C?s alkenyl , (Co - C , alkyl) C ( O )NR24C2 -C18 alkynyl, (Co - C 00, LE3 alkyl) C ( O ) NR24H2, (Co - C , alkyl) C ( O )NR24aryl , (Co - C , alkyl) C ( O )NR24heteroaryl , (Co - C , alkyl) NR24C ( 0 ) C , -C18 45 alkyl, (Co - C , alkyl) NR24C ( O )C2 - C18 alkenyl, or (Co - C , alkyl )NR24C (O )C2 - C18 alkynyl, (Co - C , alkyl) NR24C ( O ) OH , ( Co - C , alkyl )OC (O )OC , -C18 alkyl , (Co -C , alkyl) / OC ( O )OC2 - C18 alkenyl, ( Co -C , alkyl) OC (O )OC2 - C18 alkynyl , (Co - C , alkyl )OC (O )OH , (Co - C , alkyl) OC ( O )NR240 ,- ?is 50 alkyl , (Co - C , alkyl) OC ( O )NR24C2 - C18 alkenyl, (Co - Cg alkyl) OC (O )NR24C2 - C18 alkynyl, (Co - C , alkyl )OC ( O ) NR24H2, (Co - Cz alkyl) NR24 (O )OC , -Cig alkyl, (Co -Cz alkyl) OH , and NR24 ( 0 ) 0C2- C18 alkenyl, ( Co- Cg alkyl) NR24 ( 0 )0C2 - C18 alkynyl, or (Co - C , alkyl) NR24 ( O )OH ; 55 . R3 is hydrogen, (Co - C , alkyl) halo , C7 -C18 alkyl, C2- C18 alkenyl, C2 -C18 alkynyl , heteroalkyl, (Co - Cg alkyl) aryl, ( Co Cg alkyl) heteroaryl, ( Co - C , alkyl) OC - Cis alkyl, ( Co- Cg ?? alkyl )OC2 - C18 alkenyl, (Co - Cg alkyl) OC2- C18 alkynyl, (Co Cg alkyl) OH , (Co - Cg alkyl) SH , (Co - C , alkyl) NR24C , -C18 60 alkyl , (Co -C , alkyl )NR24C2 - C18 alkenyl , (Co -C , alkyl) NR24C2- C18 alkynyl, ( C . - C , alkyl) NR24H2, (Co - C , alkyl) C ( 0 ) C , - C , alkyl, (C . - C , alkyl ) C (0 )C2 - C18 alkenyl, (Co - Cg OH Ö alkyl) C (O )C2 - C18 alkynyl , (Co -C , alkyl ) C ( O )H , (Co - Cg OH . alkyl) C ( O ) aryl, (Co - C , alkyl) C ( O )heteroaryl , ( Co - C , alkyl) In some embodiments , Y acts at a glucocorticoid receptor 65 C (O )OC , -C18 alkyl, (Co - C , alkyl) C (O )OC2 - C18 alkenyl , (GR ) . In some embodiments , Y comprises any structure that (Co - C , alkyl ) C ( O )OC2 - C18 alkynyl, (Co - C , alkyl) C ( O )OH , permits or promotes agonist activity at the GR , while in (Co - C , alkyl) C ( O ) O aryl, (Co - C , alkyl) C ( O ) O heteroaryl, US 9 ,783 , 592 B2 33 34 (Co - C , alkyl )OC ( O )C , -Cg alkyl, (Co -C ,alkyl ) OC ( O )C2 - C18 ( C . alkyl) C ( O )C , -C18 alkyl, ( C , alkyl )C ( 0 )C2 - C , alkenyl, alkenyl, (Co - C , alkyl) OC (O )C2 - C18 alkynyl, (Co -C , alkyl ) (Co alkyl) C (O )C2 - C , alkynyl, (C )C (O )aryl , (C .) C (O )het C ( O )NR24C7 -C18 alkyl , (Co - C , alkyl ) C (O )NR24C2 - C18 alk - eroaryl, ( C . ) C ( O )OC ; - C , alkyl , ( C , alkyl) C ( O )OC2 - Cz alk enyl, (Co -C , alkyl) C (O )NR24C2 - C , alkynyl, (Co -C , alkyl) enyl, ( C , alkyl) C (O )OC2 - Cg alkynyl, or ( Co alkyl) C ( O ) OH ; CIONE240 cm 1lvlycNR24 milir byly 5 Rºis hydrogen or C . - C , alkyl; C ( O ) NR24H2, ( Co- C , alky1) C ( O )NR24aryl , (Co - C , alkyl) C _ 5 R9 is hydrogen or CZ - C , alkyl; ( O )NR24heteroaryl , (Co - C , alkyl) NR24C ( O ) C -C18 alkyl, R1° is hydrogen or OH ; and (Co - C , alkyl) NR24C (O )C2 - C18 alkenyl, or (Co - C , alkyl) R24 is hydrogen or CZ - C , alkyl. NR24C ( O )C2 -C18 alkynyl , ( C .- C alkyl) NR24C (O )OH , (Co For example , R2 is hydrogen or methyl; R * is hydrogen , Cg alkyl ) OC ( O ) OC - C18 alkyl, (Co - C , alkyl ) OC ( O ) OC2- fluoro , chloro , or methyl; R6 is hydrogen or C ( O ) C . - C , C18 alkenyl, (Co - C , alkyl) OC ( O ) OC2 -C18 alkynyl, (Co - Co alkyl; R7 is hydrogen , C ( O )CHz , or C ( O )CH ,CHz ; R8 is alkyl) OC ( O )OH , (Co - C , alkyl) OC (O )NR240 , -C18 alkyl, hydrogen or methyl; Rºis hydrogen or methyl; and RIO is (Co -C , alkyl) OC (O )NR24C2 - C18 alkenyl, (Co - C , alkyl )OC hydroxyl. (O )NR24C3 -C , alkynyl, (Co - C , alkyl) OC (O )NR24H ,, (Co - Nonlimiting examples of structures of Formula C include: Cg alkyl) NR24 ( 0 )OC - C18 alkyl, ( Co - Cg alkyl) NR24 (0 ) 15 OC2- C18 alkenyl, ( Co -Cz alkyl) NR24 (O )OC2 - C1g alkyny , or OH (Co -C , alkyl) NR24 (O )OH ; R™ is hydrogen , C7- C18 alkyl, C2 -C18 alkenyl, C2- C18 alkynyl, heteroalkyl , (Co - C , alkyl ) aryl, (Co - C , alkyl) het Me eroaryl , ( C . - C . alkyl) C ( O ) C -C18 alkyl, ( Co- C , alkyl) C ( O ) ?? 11111OH , C2- C18 alkenyl, (Co -Cg alkyl ) C ( O ) C2 -Cis alkynyl, (Co - Cg 20 Me alkyleroaryl ) C , ( O( Co ) H - , C( Co , alky1 - C , alkyl) C (O ) ) COC ( O ,)- arylC18 , (alkyl Co- Cg, (Coalkyl - C) , C alkyl( O )het ) C (O )OC2 -C18 alkenyl, (Co - C , alkyl )C (O )OC2 - C18 alkynyl, (Co -C , alkyl) C (O )OH , C . - C , alkyl) C (O ) O aryl , (Co - C , alky1 ) C (O ) O heteroaryl, (Co - C , alky1 ) C ( O )NR240 , - C18 25 alkyl, (Co - Cz alkyl) C (O )NR24C2 - C18 alkenyl, (C . - C , alkyl) Cortisol C ( O )NR24C2 -C18 alkynyl , ( C . - C , alkyl) C ( O )NR24H2 , (Co Co alky1) C (O )NR24 aryl , or (Co - C , alkyl )C ( O ) NR24heteroaryl; Hc R ? is hydrogen , C , - C18 alkyl, C2 -C18 alkenyl, C2- C18 30 alkynyl, heteroalkyl, (Co - C , alkyl) aryl, (Co - C , alkyl )het eroaryl, (Co - C , alkyl) C ( O ) C -C18 alkyl, ( C . - C . alkyl) C ( O ) C2- Cig alkenyl , ( Co -Cg alkyl) C (0 )C2 - C18 alkynyl, (Co - C , Me alkyl) C (OH , ( C . -Cz alkyl) C ( O ) aryl , (Co - Cz alkyi ) C ( O )het HO fa .. . 1111 OH , eroaryl, (Co - C , alkyl) C ( O )OC , -C18 alkyl, (Co - C , alkyl ) C 35 Me ( 0 )0C2 - C18 alkenyl, (Co - C , alkyl) C ( O )OC2 - C18 alkynyl, (Co -C , alkyl) C ( O )OH , C . - C , alkyl) C (O ) O aryl, (Co - C , alkylalky1, ) C( co ( O - ) C O , alky1heteroaryl) C (O )NR24C2, (Co - C- Cig, alkyl alkenyl) C ( O, )(NR240 Co -C , alkyi, -C18 ) C (O )NR24C2 - C18 alkynyl, (Co -C , alkyl )C ( O ) NR24H2, (Co Co alkyl) C ( O )NR24 aryl , or (Co - Cg alkyl) C (O ) Cortisone acetate NR24heteroaryl; R8 is hydrogen , (Co -C , alkyl) halo , C7 -C18 alkyl , C2 -C18 alkenyl, C2- C18 alkynyl, heteroalkyl, (Co -Cg alkyl) aryl, (Co Et Cg alkyl) heteroaryl ; Rº is hydrogen , (Co -C , alkyl) halo , C ,- C18 alkyl, C2- C18 45 alkenyl, C2 -C18 alkynyl , heteroalkyl, (Co - C , alkyl) aryl, (Co O Cg alkyl) heteroaryl ; Me HO 11110 RlO is hydrogen , (Co - C , alkyl) halo , C . - C18 alkyl, or ( Co Et, Cg alkyl )OH ; and Me H 11111CH3 R24 is hydrogen or C , -C18 alkyl. 50 In some embodiments , Y comprises a structure of For mula C , wherein R2 is hydrogen , halo , OH , or C , - C , alkyl; R® is hydrogen , halo , OH , or C - C , alkyl; R is hydrogen , C , -C18 alkyl, C2- Cg alkenyl , C2 - C , alky - 55 Beclometasone nyl, heteroalkyl, (Co - C , alkyl) aryl, (Co - C , alkyl )heteroaryl , OH (Co -C , alkyl) C ( O ) C -C18 alkyl, (Co - C , alkyl) C (O )C2 - Cg alkenyl, (Co - C , alkyl) C ( O )C2 - C , alkynyl, (Co - C , alkyl) C (O )H , (C .- C , alkyl )C ( O ) aryl, (Co - C , alkyl) C (O )heteroaryl , Me (Co - C , alkyl) C (O )OC , -C , alkyl, (Co -C , alkyl) C (O )OC2 - C , ó MOH alkenyl, (Co -C , alkyl) C (O )OC2 - C , alkynyl, (Co -C , alkyl) C _ 00 (O )OH , C . -C , alkyl) C (O ) O aryl, (Co -C , alkyl) C (O )O het .. ! ! 111CH3, eroaryl, (Co - C , alkyl) C ( O )NR24C , -C , alkyl , (Co - Cz alkyl) C II.) (O )NR24C2 - C , alkenyl, (Co -C , alkyl) C (O )NR24C2 - C , alkynyl , Co- C , alky1) C ( O )NR24H ,, ( -Cz alkyl) C ( O ) NR24aryl, or (Co - C , alkyl) C (O )NR24heteroaryl ; 65co R ? is hydrogen , C . - C . alkyl, C2- Cg alkenyl, C2- C , alky Prednisone nyl , heteroalkyl, (Co - C , alkyl) aryl, (Co - C , alkyl )heteroaryl , US 9 ,783 ,592 B2 35 36 - continued conjugation on Formula C and means of conjugation of OH Formula C to Q or L in view of general knowledge and the disclosure provided herein . In some embodiments , Formula C is conjugated to L or Q at any of positions 1 , 2 , 3 , 4 , 5 , 6 , Me 5 7 , 8, 9 , 10 , 11 , 12 , 13 , 14 , 15 , 16 , 17 , 18 , 19, 20 , 21 , 22 , or HO OH , 23 of Formula C . In some embodiments , Formula C is Me conjugated to L or Q at position 3 , 10 , 16 or 17 of Formula In some embodiments , Y acts at a mineralcorticoid recep ?IH 10 tor MR( ) . In some embodiments , Y comprises any structure that permits or promotes agonist activity at the MR , while in other embodiments Y is an antagonist of MR . In exemplary Prednisolone embodiments , Y comprises a structure of Formula D : OH Formula D O - R7

Me Me HO cutOH . R 10 Me 20 H CORITI . 07 CH3 25 wherein R2, R3, R7 and R10 are each independently a moiety Methylprednisolone that permits or promotes agonist or antagonist activity upon OH binding of the compound of Formula D to the MR ; and the dashed line indicates an optional double bond . In some embodiments , Formula D further comprises one or more Me 30 substituents at one or more of positions 1 , 2 , 4 , 5 , 6 , 7 , 8 , 11 , HO 2011OH 12 , 14 , 15 , 16 , and 17 . In some embodiments , Y comprises a structure of For Me CH3, mula D wherein R² is hydrogen , (Co - C , alkyl) halo , C7 -C18 alkyl, C2- C18 Tin 35 alkenyl , Cz- C alkynyl, heteroalkyl , (Co - C , alkyl) aryl , ( Co Cg alkyl) heteroaryl , (Co - Cg alkyl) OC , -Cig alkyl , (Co - C 0, alkyl )OC2 - C18 alkenyl, (Co - C , alkyl) OC2 - C18 alkynyl , (Co Betamethasone Cg alkyl) OH , ( C -C , alkyl) SH , (Co - C , alkyl) NR240 , -C1s OH alkyl, ( Co- Cg alkyl )NR24C2 - C18 alkenyl , (Co - C , alkyl) NR24C2- C18 alkynyl, (Co -Co alkyl) NR24H2 , (Co -Co alkyl) C 40 (0 ) C -C18 alkyl, ( C - C alkyl ) C (O )C2 -C18 alkeny? , ( Co - C , Me FO alkyl ) C ( O )C2 -C18 alkynyl , (Co - C , alkyl) C ( O ) H , (Co - Cg HOHOHHO .. . ??? alkyl) C (O aryl, (Co - Cg alkyl) C ( O ) heteroaryl, ( Co- Cg alkyl ) C ( O )OC , -C18 alkyl, (Co - C , alkyl) C ( O ) OC2- C18 alkenyl, Me HLYH 111111 OH , (Co - C , alkyl) c (o )0C2 - C18 alkynyl, (Co - Cz alkyl) C ( O )OH , 45 (Co -Co alkyl) C (O ) O aryl, (Co - C , alkyl) C ( O )O heteroaryl, | IM (Co - C , alkyl) oC (O ) c ,- C18 alkyl, ( Co - C , alkyl) OC ( O )C2 C18 alkenyl, (Co - C , alkyl) OC ( O )C2 - C18 alkynyl , (Co - C , alky1) C ( O )NR240 , -C18 alkyl, (Co - C , alkyl) C ( O ) NR24C2 Triamcinolone Cig alkenyl, ( Co - C , alkyl ) C (O )NR24C2 - C18 alkynyl , ( Co- Cg OH alkyl ) C ( O )NR24H , (Co - C , alkyl) CONR24aryl , (Co - C 00 alkyl) C ( O )NR24heteroaryl , (Co - C , alkyl) NR24C ( 0 ) C , -C18 alkyl , (Co - C , alkyl) NR24CO ) C -C18 alkenyl , or (Co - C , alkyl) NR24C ( O )C2 - C18 alkynyl , (Co -C , alkyl) NR24C (O ) HO JanuOH OH , ( C . - C , alkyljoC ( O )OC -C18 alkyl, (Co - C , alkyl )O? ( O )OC2 - C18 alkenyl, (Co - C , alkyl )OC ( O )OC2 - C18 alkynyl , Me 11111CH3, 55 ( Co- Co alkyl) OC ( O )OH , ( C . - C alkyl) OC ( O )NR240 , - C18 alkyl, (Co - C , alkyl) OC ( O )NR24C2 - C18 alkenyl , (Co - C , | alkyl) OC ( O )NR24C2 - C18 alkynyl , (Co - C , alkyl )OC ( O ) NR24H2, (Co -Cg alkyl) NR24 (O )OC , -Cig alkyl, (Co -Cg alkyl) NR24 (O )OC2 - C18 alkenyl , ( Co- C , alkyl) NR24 (O )OC2 - C18 Dexamethasone 60 alkynyl, or (Co - C , alkyl) NR24 (O )OH ; R3 is hydrogen , (Co - C , alkyl) halo , C ,- C18 alkyl, C2 -C18 and derivatives thereof. alkenyl, C2- C18 alkynyl, heteroalkyl , (Co -Cg alkyl) aryl , (Co In embodiments wherein Y comprises a structure of Cg alkyl) heteroaryl , (Co - C , alkyl )OC - C18 alkyl, (Co - C8 Formula C , Y is conjugated to L ( e .g . when L is a linking alkyl) OC2 - C18 alkenyl , (Co - Cg alkyl) OC2 -C18 alkynyl, (Co group ) or Q (e . g. when L is a bond ) at any position of 65 Cg alkyl) OH , (Co - C , alkyl) SH , (Co - C , alkyl) NR24C , -C18 Formula C that is capable of reacting with or L . One alkyl, (Co - C , alkyl) NR24C2 - C18 alkenyl, (Co -C , alkyl) skilled in the art could readily determine the position of NR24C2 -C18 alkynyl, (Co - Cg alkyl) NR24H2 , (Co -C , alkyl) C US 9 ,783 , 592 B2 37 38 (0 )C , - C , alkyl, (C . -C , alkyl )C ( O ) C2- C18 alkenyl, ( Co- C , alkyl ) C ( O )C2 -C18 alkynyl , (Co - C , alkyl) C ( O ) H , ( C . - C . -continued alkyl) C ( O ) aryl, ( Co- C , alkyl ) C ( O )heteroaryl , (Co - C , alkyl) C (O )OC -C18 alkyl , (Co - C , alkyl ) C ( O ) OC2- C18 alkenyl, (Co - Cz alkyl ) C ( O )OC2 - C18 alkynyl, (Co - C , alkyl) C ( O )OH , 5 (C . -C . alkyl) C (O ) O aryl, (Co -Cz alkyl) C ( O )O heteroaryl , (Co -C , alkyl) OC (O ) C , - C18 alkyl, (Co -Cg alkyl) OC ( O )C2 Cis alkenyl, (C . - C , alkyl )OC (O ) C - Cis alkynyl, (Co -Cg HO Me alkylCigalkenyl) C ( O ) NR24C7, ( C . - C . - C18alkyl alkyl) C ( O, ) NR24C2 (Co - C , -alkyl C s alkynyl) C (O )NR24C2 , ( C . - C . alkyl ) C ( O )NR24H2 , (Co - C , alkyl) C ( O )NR24aryl , ( C . - C alkyl) CO )NR24heteroaryl , (C . - C , alkyl) NR24 _ ( O ) C -C18 ST alkyl , (Co - C , alkyl )NR24C ( O )C2 - C18 alkenyl, or (Co -Cg alkyl) NR24C ( O )C2 - C18 alkynyl , (Co - C , alkyl )NR24C ( ) OH , (Co - C , alkyl oC ( O )OC , -C18 alkyl, (Co - C , alkyl) OC Deoxycorticosterone acetate ( O )OC2 - C18 alkenyl , ( Co - Cz alkyl )OC ( O )OCZ - C18 alkynyl , 15 (Co - Cg - alkyl) OC (O )OH , ( Co - C , alky? )OC ( O NR240 , -C18 alkyl, (Co -C , alkyl) OC (O )NR24C2 -C18 alkenyl , ( C .- C alkyl) OCONR24C2 - C18 alkynyl, (Co - C alkyl) OC ( O ) NR24H2, (C . -C , alkyl) NR24 ( O )OC , -C18 alkyl, (Co -C , alkyl ) Me NR24 ( O )OC2 - C18 alkenyl, (Co -Cg alkyl) NR24 (O )OC2 - C18 20 HON alkynyl , or (Co - C , alkyl) NR24 (O )OH ; R ? is hydrogen , C7 -C18 alkyl, C2- C18 alkenyl, C2- C18 Me alkynyl, heteroalkyl , (Co - C , alkyl ) aryl, (Co - C , alkyl) het H eroaryl, (Co - C , alkyl) C ( 0 ) C , -C18 alkyl, (Co - C , alkyl) C ( O ) | C2- C18 alkenyl , (Co - C , alkyl ) C ( O )C2 -C18 alkynyl, (Co - C , 25 alkyl) C ( O ) H , (Co - C , alkyl) C ( O ) aryl, (Co -Cz alkyl) C ( O )het TE eroaryl, ( C .- C , alkyl) C (O )OC ,- C18 alkyl, (C . -C , alkyl) C ( O )OC2 - C18 alkenyl, (Co - C , alkyl) C ( O )OC2 - C18 alkynyl , Deoxycorticosterone acetate (Co - C , alky1 ) C ( O )OH , Co- C , alkyl) C ( O ) O aryl , (Co - C , alkyl) C (O )O heteroaryl, ( C . - C alkyl) C ( O )NR24C ,1 -418C18 30 and derivatives thereof. alkyl, (Co - C , alkyl) C ( O )NR24C2 - Cig alkenyl, (C . - C , alkyl) 30 In embodiments wherein Y comprises a structure of C ( O )NR24C2 -C18 alkynyl, (C . - C , alkyl) C ( O )NR24H2 , (Co Formula D , Y is conjugated to L ( e . g . when L is a linking NR24heteroarylCg alkyl) C ( O; )NR24aryl , or (Co -C , alkyl) C (O ) group ) or Q ( e . g . when L is a bond ) at any position of Rlº is hydrogen , ( Co - C , alkyl )halo , C . -C18 alkyl , or (Co Formula D that is capable of reacting with Q or L . One Cg alkyl) OH ; and 35 skilled in the art could readily determine the position of R24 is hydrogen or C1- C18 alkyl. conjugation on Formula D and means of conjugation of In some embodiments , Y comprises a structure of For Formula D to Q or L in view of general knowledge and the mula D , wherein disclosure provided herein . In some embodiments , Formula R² is hydrogen , halo , OH , or C , - C , alkyl; D is conjugated to L or Q at any of positions 1 , 2, 3, 4 , 5 , R * is hydrogen , halo , OH , or C7 -C , alkyl; R ' is hydrogen , C . - C , alkyl, C2- C , alkenyl , C2- C , alky 40 6 , 7 , 8 , 9 , 10 , 11 , 12 , 13, 14 , 15, 16 , 17, 18 , 19, 20 , 21, 22 , nyl, heteroalkyl , (Co - C , alkyl) aryl , (Co - C , alkyl) heteroaryl, 23 , or 24 of Formula D . In some embodiments , Formula D (Co alkyl) C ( O ) C - C alkyl, (Co alkyl) C ( O )C2 - C , alkenyl, is conjugated to L or Q at position 3 , 10 , 13 , or 17 of Formula (C . alkyl) C (O )C2 - C , alkynyl , (C .) C ( O ) aryl, (C .) C ( O )het D . eroaryl , ( C . ) C ( O ) OC - Cg alkyl, ( C , alkyl) C ( O )OC2 - Cg alk - 45 In some embodiments , Y acts at a progesterone receptor enyl, ( C , alkyl ) C (O )OC2 - C , alkynyl, or (C , alkyl) C ( O )OH ; * (PR ) . In some embodiments , Y comprises any structure that R10 is hydrogen or OH ; and permits or promotes agonist activity at the PR , while in other R24 is hydrogen or C . -C , alkyl. embodiments Y is an antagonist of PR . In exemplary For example , R2 is hydrogen or methyl; R² is hydrogen , embodiments , Y comprises a structure of Formula E : fluoro , chloro , or methyl; R ' is hydrogen , C (O )CH3 , or 50 C ( O )CH2CH3 ; and R10 is hydroxyl. Nonlimiting examples of compounds of Formula D Formula E include: O 0 - R ? use OH .

HO 12 Me g wherein R ?, R3, R4, and R7 are each independently moieties that permit or promote agonist or antagonist activity upon 65 binding of the compound of Formula E to the PR ; and the Aldosterone dashed line indicates an optional double bond . In some embodiments , Formula E further comprises one or more US 9 ,783 , 592 B2 39 40 substituents at one or more of positions 1 , 2 , 4 , 5 , 6 , 7 , 8 , 11 , C , alkyl) OH , (Co - C , alkyl) SH , (Co -C , alkyl) NR24C7 - C18 12 , 14 , 15 , 16 , and 17 ( e . g . a methyl group at position 6 ). alkyl, (Co - Cg alkyl) NR24C2 - C18 alkenyl, (Co - Cg alkyl) In some embodiments , Y comprises a structure of For NR24C2- C18 alkynyl, (Co -Cg alkyl )NR24H2 , (Co - C , alkyl ) C mula E wherein ( 0 ) C - C , alkyl , (Co - C , alkyl) C ( O )C2 - C18 alkenyl, ( Co - C8 R² is hydrogen , (Co - C , alkyl) halo , C1 -C18 alkyl , C2- C18 5 alkyl) C ( O )C2 - C18 alkynyl , (Co - C , alkyl) C ( O ) H , (Co - Cg alkenyl, C2- C18 alkynyl, heteroalkyl , (Co -Cg alkyl) aryl, (Co - alkyl) C ( O )aryl , (Co - C , alkyl) C ( O )heteroaryl , (Co - C , alkyl) C , alkyl) heteroaryl , ( Co - C , alkyl) OC - C18 alkyl, (Co -C8 C ( O ) OC , - C18 alkyl, (Co - C , alkyl) C ( O ) OC2- C18 alkenyl, alkyl) OC2 -C18 alkenyl, (Co - C , alkyl) OC2 -C18 alkynyl, ( C . - (Co - C , alkyl ) C ( O )OC2 - C18 alkynyl, (Co - C , alkyl) C (O )OH , Cg alkyl )OH , (Co - C , alkyl )SH , (Co -C , alkyl) NR24C7 - C18 (Co -C , alkyl) C (O )O aryl, (Co - C , alkyl) C ( O )O heteroaryl, alkyl, ( C . - C . alkyl) NR24C2 - C18 alkenyl, (Co - C , alkyl ) 10 (Co - C alkyl) OC ( O ) C - C , alkyl, (Co - Cz alkyl) OC (O )C2 - C18 NR24C2 -C18 alkynyl, ( Co -Cg alkyl) NR24H2 , (Co - C , alkyl) C alkenyl, (Co - C , alkyl) OC ( O )C2 - C18 alkynyl, (Co - Cg alkyl) (O ) C -C18 alkyl, (Co - C , alkyl) C (O )C2 - C18 alkenyl, (Co - Cg C ( O )NR24C7 -C18 alkyl, ( Co - C , alkyl) C ( O )NR24C2 - C18 alk alkyl) C ( O )C2 - C18 alkynyl, (C . - C , alkyl) C ( O )H , (Co - C , enyl, (Co - C , alkyl) C ( O )NR24C2 - C18 alkynyl, ( C - C , alkyl) alkyl) C ( O )aryl , (Co - C , alkyl) C (O ) heteroaryl, (Co -C , alkyl) C (O )NR24H2 , (Co -C , alkyl) C ( O )NR24aryl , (Co - C , alkyl) C C ( O ) OC , -C18 alkyl , (Co - C , alkyl ) C ( O )OC2 - C18 alkenyl, 15 ( O )NR24heteroaryl , (Co - C , alkyl) NR24C ( O ) C , -C18 alkyl, (Co - C , alkyl )C (O )OC2 - C18 alkynyl, (C . -C , alkyl) C (O )OH , (Co - C , alkyl) NR24C (O )C2 - C18 alkenyl , or (Co -Cg alkyl ) ( Co- C , alkyl )C ( O ) O aryl, (Co - C , alkyl ) C (0 ) heteroaryl, NR24C ( O )C2 - C18 alkynyl, ( Co - C , alkyl) NR24C ( O )OH , (Co (C .- C , alkyl )OC ( O )C , -C18 alkyl, (Co - C , alkyl )OC (O )C2 Cg alkyl) OC (O ) OCZ- C18 alkyl, (Co -C , alkyl )OC (O )OC2 C18 alkenyl , (Co - C , alkyl) OC (O )C2 - C18 alkynyl , (Co - C8 C18 alkenyl , (Co - C , alkyl )OC ( O ) OC2- C18 alkynyl, (Co - C8 alkyl) C (O )NR24C7 -C18 alkyl , (Co -C , alkyl) C ( O )NR24C2 - 20 alkyl) OC (O )OH , ( Co - C , alkyl) OC ( O )NR24C7 - C18 alkyl, Cis alkenyl, ( C - C alkyl) C ( O )NR24C2 -C18 alkynyl, ( C . - C . (Co - C , alkyl) OC ( O )NR24C2 - Cis alkenyl, (Co - C , alkyl )OC alkyl) C ( O )NR24H2 , (Co - C , alkyl) C (O )NR24aryl , ( C .- C . (O )NR24C2 -C18 alkynyl , (Co - C , alkyl) OC (O )NR24H2 , (Co alkyl) C ( O )NR24heteroaryl , ( Co - C , alkyl) NR24C (O ) C - Cis C , alkyl) NR24 (O )OC - C18 alkyl , (Co - C , alkyl) NR24 ( 0 ) alkyl, (Co - C , alkyl )NR24C ( O )C2 - C18 alkenyl, or (Co - C , OC2- C18 alkenyl, (Co - Cg alkyl) NR24 (O )OC2 - C18 alkynyl, or alkyl )NR24C ( O ) C2- C18 alkynyl, (Co - C , alkyl) NR24C ( O ) 25 (Co - C , alkyl )NR24 (O )OH ; OH , (C .- C , alkyl )OC ( O )OC , -C18 alkyl, (Co -Cg alkyl) OC R ’ is hydrogen , C1- C18 alkyl , C2- C18 alkenyl , C2- C18 ( O )OC2 - C18 alkenyl, ( C . - C . alkyl) OC ( O ) OC2- C18 alkynyl, alkynyl, heteroalkyl , (Co - C , alkyl) aryl, ( C . - C , alkyl) het ( C . - C , alkyl) OC ( O )OH , ( C . - C , alkyl) OC ( O )NR24C , -C18 eroaryl, (Co - Cg alkyl) C ( O )C7 -C18 alkyl, (Co - C , alkyl) C ( O ) alkyl, (Co - C , alkyl )OC (O )NR24C2 - C18 alkenyl, ( Co- Cg C2 -C18 alkenyl, (Co - C , alkyl )C ( 0 )C2 -C18 alkynyl, (Co -Cg alkyl) OC ( O )NR24C2 - C18 alkynyl, (Co - C , alkyl) OC ( O ) 30 alkyl) C ( O ) H , ( C . - C . alkyl) C ( O ) aryl, (Co - Cg alkyl ) C ( O )het NR24H2, (Co -Cg alkyl) NR24 (O )OC , -C18 alkyl , ( C . -C alkyl) eroaryl , (C . -C , alkyl) C ( O )OC , -C18 alkyl, (Co - C , alkyl )C NR24 ( O ) OC2 -C18 alkenyl , (Co - C , alkyl) NR24 (O )OC2 - C18 ( O )OC2 - C18 alkenyl, (Co - C , alkyl) C ( O ) OC2- C18 alkynyl, alkynyl, or (Co - C , alkyl )NR24 ( O )OH ; R24 is hydrogen or (Co - C , alkyl) C ( O )OH , C . - C , alkyl) C ( O ) O aryl, (Co - C , C - C S alkyl. alkyl) C ( 0 ) 0 heteroaryl, (Co -C , alkyl) C (O )NR24C , - C18 R3 is hydrogen , (Co -Cz alkyl) halo , C . -C18 alkyl, C2- C18 35 alkyl, (Co - Cz alkyl) C ( O )NR24C2 - C18 alkenyl, (Co - Cz alkyl) alkenyl , C2- C18 alkynyl, heteroalkyl , (Co - Cg alkyl) aryl, (Co - C ( O )NR24C2 -C18 alkynyl, (Co -Cg alkyl) C ( O )NR24H2 , (Co C , alkyl) heteroaryl , (Co - C , alkyl )OC , - C18 alkyl, (Co - C , Cg alkyl) C ( O )NR24aryl , or (Co -C , alkyl) C (O ) alkyl) OC2 -C18 alkenyl, (Co - C , alkyl) OC2 -C18 alkynyl, (Co - NR24heteroaryl ; and Cg alkyl )OH , (Co - C , alkyl ) SH , (Co -C , alkyl) NR24C , - C18 R24 is hydrogen or C , - C18 alkyl. alkyl, (Co - C , alkyl) NR24C2 - C18 alkenyl, (Co - C , alkyl) 40 In some embodiments , Y comprises a structure of For NR24C2- C18 alkynyl, (Co - C , alkyl) NR24H2 , (Co - C , alkyl )C mula E , wherein ( O ) C , - C , alkyl, ( Co- Cg alkyl) C ( O )C2 - C18 alkenyl, ( Co- C8 R² is hydrogen, halo , OH , or CZ - C , alkyl ; alkyl) C ( 0 ) C2- C18 alkynyl, (Co - C , alkyl) C ( O ) H , (Co -Co Rºis hydrogen , halo , OH , or CZ - C , alkyl; alkyl ) C ( O )aryl , (Co - C , alkyl ) C ( O ) heteroaryl , (Co - C , alkyl) R4 is hydrogen , (Co - C , alkyl) halo , C7 -C18 alkyl, C2- C , C ( O )OC -C18 alkyl, (Co -C , alkyl) C ( O ) OC2 -C18 alkenyl, 45 alkenyl, C2- 18 alkynyl, heteroalkyl, (C .- C , alkyl) aryl, (Co ( Co- Cg alkyl) C ( O ) OC2 -C18 alkynyl, (Co - C , alkyl) C ( O ) OH , Cg alkyl) heteroaryl , (Co - C , alkyl )0C1 - C , alkyl, (Co -Cg

( C - C , alkyl) C (O ) O aryl , (Co - C , alkyl) C ( O )O heteroaryl, alkyl )OC2 - Cg alkenyl, (Co - Cg alkyl) OC2 - C , alkynyl, (Co - C O, (Co - C , alkyl) OC ( O ) C , - C18 alkyl, (Co -Cg alkyl) OC ( O ) C2 211alkyl, ) OH , (Co - C , alkyl) SH , ( C .- C , alkyl )NR24C7 - C , alkyl , C18 alkenyl, (Co - C , alkyl) OC ( O )C2 -C18 alkynyl, (Co - Cg2 (Co - C , alkyl)NR24C2 - C , alkenyl, (Co - C , alkyl) NR24C2- C , alkyl) C ( O )NR24C7 -C18 alkyl, (Co - C , alkyl )C (O )NR24C2 - 50 alkynyl, (Co - Cg alkyl) NR24H2 , (C .- C , alkyl) C (0 )C -Cg C18 alkenyl , (Co - C , alkyl) C (O )NR24C2 -C18 alkynyl , ( Co- Cg alkyl , (Co - C , alkyl) C ( O ) C2- Cg alkenyl , (Co - C , alkyl) C ( O ) alkyl) C ( O )NR24H2 , (Co - C , alkyl) C (O )NR24 aryl, (Co -C , C2- Cg alkynyl , (Co - C , alkyl) C (O )H , (Co - Cg alkyl ) C (O ) aryl, alkyl) C ( O )NR24heteroaryl , (Co - C , alkyl )NR24C ( O )C7 -C18 (Co - C , alkyl) C ( O ) heteroaryl, (Co - C , alkyl) C ( O ) OCZ - C , alkyl, (Co - C , alkyl) NR24C ( 0 ) C2- C18 alkenyl , or (Co - C , alkyl, (C . - C . alkyl) C (O )OC2 - C , alkenyl , (Co -Cz alkyl) C (O ) alkyl) NR24C (O )C2 - C18 alkynyl, (Co - C , alkyl) NR24C (O ) 55 OC2- C , alkynyl, (Co - C , alkyl) C (O )OH , (Co -C , alkyl) C OH , (C .- C , alkyl )OC (O )OC , -C18 alkyl , (C . -C , alkyl )OC (0 ) 0 aryl, (Co -C , alkyl) C (O ) O heteroaryl, ( C .- C alkyl) OC (O )OC2 - C18 alkenyl , (Co - C , alkyl) OC ( O )OC2 - C18 alkynyl, (0 )C - C alkyl, (Co - C , alkyl) OC ( O )C2 - Cg alkenyl, (Co -C , ( Co- C , alkyl) OC ( O )OH , (Co - C , alkyl) OC ( O )NR24C7 -C18 alkyl )OC ( O ) C2- C18 alkynyl , ( C . - C , alkyl ) C ( O ) NR24C , - C , alkyl, (Co - C , alkyl) OC ( O )NR24C2 -C18 alkenyl , (Co - C , alkyl, (Co - C , alkyl) C ( O )NR24C2 - C , alkenyl, (Co - C , alkyl ) alkyl )OC ( O )NR24C2 - C18 alkynyl, (Co - C , alkyl) OC ( O ) 60 C ( O )NR24C2 - C , alkynyl, (Co - C , alkyl) C ( O )NR24H2 , (Co NR24H , (Co -Cz alkyl) NR24 ( O )OC , - C18 alkyl, (Co - C , alkyl) Cg alkyl) C ( O ) NR 24 aryl, ( C . - C , alkyl) C ( O ) NR24heteroaryl, NR24 (O )OC2 - C18 alkenyl, (Co -C , alkyl) NR24 (O )OC2 -C18 (C . -C , alkyl )NR24C (0 )C , -C , alkyl, (Co - C , alkyl )NR24C alkynyl, or (Co -C , alkyl) NR24 (O )OH ; (0 )C2 - C , alkenyl, or (Co - C , alkyl) NR24C (O )C2 - C , alkynyl , R * is hydrogen , ( Co- C , alkyl) halo , C . - C18 alkyl, C2- C18 (Co - C , alkyl )NR24C ( O )OH , (Co - C , alkyl) OC ( O ) OC . - C alkenyl, C2 -C18 alkynyl , heteroalkyl, (Co - Cg alkyl) aryl , (Co - 65 alkyl , (Co -C , alkyl) OC (O )OC2 -Cg alkenyl, (Co - C , alkyl) OC Cg alkyl) heteroaryl , (Co - C , alkyl )OC ,- C18 alkyl, ( Co- Cg ( O )OC2 - C , alkynyl, (C -C , alkyl) OC (O ) OH , (Co - C , alkyl) alkyl )OC2 - C1g alkenyl, (Co -C , alkyl) oC2 - C18 alkynyl, (Co - OC ( O )NR240 , -C , alkyl, (Co - Co alkyl )OC (O )NR24C2 -C US 9 ,783 , 592 B2 4 ) 42 alkenyl, (Co - C , alkyl) OC ( O ) NR24C2- C , alkynyl, (Co -C8 or 24 of Formula E . In some embodiments , Formula E is alkyl) OC (O )NR24H2 , (Co -Cz alkyl) NR24 (O )OC , - C , alkyl, conjugated to L or Q through position 3 or 17 of Formula E . (Co - C , alkyl )NR24 (0 )OCZ - C , alkenyl, (Co -C , alkyl) NR24 In other embodiments , Y acts at a progesterone receptor ( O )OC2 - C , alkynyl , or ( Co- C , alkyl) NR24 (O )OH ; but is not is not encompassed by Formula E . For example , R ' is hydrogen , C , -C , alkyl, C2 -C , alkenyl, C2 -C , alky - 5 Y can comprise the below structure and analogs thereof: nyl, heteroalkyl, (Co - C , alkyl) aryl, (Co -Cg alkyl )heteroaryl , ( C . alkyl )C (O ) C , -C , alkyl, ( C , alkyl) C ( O )C2 - Cg alkenyl, ( C . alkyl) C ( O )C2 - C , alkynyl , ( C . ) C ( O ) aryl, ( C . ) C ( O )het Me OH eroaryl, (C ) C ( O )OC - C alkyl, (C . alkyl) C ( O )OC2 - Cg alk = enyl, (C , alkyl) C (O )OC2 - C , alkynyl, or (Co alkyl )C (O ) OH ; and H R24 is hydrogen or C1- C , alkyl. For example , R2 is hydrogen or methyl; R2 is hydrogen or INTR methyl; R * is (C , alkyl) C (O ) C , - C4 alkyl, acetate , cypionate , 15 hemisucciniate , enanthate, or propionate ; and R ’ is hydro gen , C ( O ) CHz, or C ( O )CH2CH3 . Norethindrone Nonlimiting examples of compounds of Formula E include: In some embodiments , Y acts at an androgen receptor 20 ( AR ) . In some embodiments , Y comprises any structure that permits or promotes agonist activity at the AR , while in other embodiments Y is an antagonist of AR . In exemplary CH3, Me embodiments , Y comprises a structure of Formula F : 25 Me Formula F (RO ) INI.) Me

Progesterone TI

- CH3, Me (Rhod 35 H wherein R ', when present , R², R3 and R? are each indepen dently a moiety that permits or promotes agonist or antago i nist activity upon binding of the compound of Formula F to the AR ; and each dashed line represents an optional double 40 bond , with the proviso that no more than one of the optional carbon -carbon double bond is present at position 5 . In some 19 -nor - progesterone embodiments , Formula F further comprises one or more substituents at one or more of positions 1 , 2 , 4 , 5 , 6 , 7 , 8 , 11 , 12 , 14 , 15 , 16 , and 17 . Me 45 In some embodiments , Y comprises a structure of For "! ! 1111 mula F wherein Me ? ?H R ! is present and is hydrogen , C .- C18 alkyl, C2- C18 CH3 alkenyl, C2- C18 alkynyl, heteroalkyl, (Co -Cg alkyl) aryl, (Co Cz alkyl )heteroaryl , ( Co - C , alkyl) C ( O ) ? , -C18 alkyl, (Co - Ca IH 50 alkyl ) C ( O )C2 - C18 alkenyl, (Co - C , alkyl) C ( O )C2 - C18 alky nyl , (Co - C , alkyl) C ( O ) H , (Co - C , alkyl ) C ( O )aryl , (Co - Cg alkyl) C ( O ) heteroaryl , ( C . - C alkyl) C ( O ) OC , -C18 alkyl , (Co - C , alky1) C (O )OCZ - Cig alkenyl, (Co -C alkyl) C ( O )O? , Cig alkynyl, ( Co- Cg alkyl) C ( O )OH , Co - C , alkyl) C ( O ) O aryl, Medroxyprogesterone 55 (Co - Cg alky1 )C (O )O heteroaryl, (Co - C , alkyl) C ( O )NR240 , Cis alkyl, (Co -C , alkyl )C (O )NR24C2 -C18 alkenyl, (Co - C , alkyl) C ( O )NR24C2 -C18 alkynyl , ( C . - C . alkyl) C ( O )NR24H2 , and derivatives thereof. (Co - C , alkyl ) C (O )NR24aryl , (Co -C alkyl )C (O ) In embodiments wherein Y comprises a structure of NR24heteroaryl, or SOZH ; Formula E , Y is conjugated to L (e . g . when L is a linking R2 is hydrogen . ( C . - C . alkyl halo . C . - C . alkyl. C . - C . group ) or Q ( e . g . when L is a bond ) at any position of 60 alkenvl. C . - C . alkynyl, heteroalkyl. ( C . - C . alkyljaryl, ( C . Formula E that is capable of reacting with Q or L . One 116 skilled in the art could readily determine the position of alkylCg alkyl OC) .heteroaryl - C . alkenyl, (,Co ( C- . C -, C .alkyl alkyl) OC ) OC - .Cis - C . alkylalkynyl, ( C, .( - C C. , conjugation on Formula E and means of conjugation of C , alkyl OH , ( C . - C . alkyl ) SH , (C . - C , alkyl) NR24C , - C , Formula E to Q or L in view of general knowledge and the alkyl, (Co - C , alkyl) NR24C , - C , alkenyl, (C . -C , alkyl) disclosure provided herein . In some embodiments , Formula 65 NR24C2 -C18 alkynyl, (Co - C , alkyl) NR24H2, (Co - C , alkyl) C E is conjugated to L or Q at any of positions 1, 2, 3 , 4 , 5 , 6 , ( O ) C -C18 alkyl, (C .- C , alky1 )C (O )C2 - C18 alkenyl, (Co - C . 7 , 8 , 9 , 10 , 11, 12 , 13 , 14 , 15 , 16 , 17 , 18 , 19, 20 , 21, 22, 23 , alkyl) C ( O )C2 - C18 alkynyl, (Co -C , alkyl) C ( O )H , (Co - Cg US 9 ,783 , 592 B2 43 44 alkyl) C ( O ) aryl, (Co - C , alkyl) C ( O )heteroaryl , ( C . - C , alkyl) R is hydrogen , halo , OH , or CZ- C , alkyl; C ( O )OC , -C18 alkyl, (Co - C , alkyl) C (O )OCZ - Cig alkenyl, Rº is hydrogen , C , - C , alkyl, Cz - C , alkenyl, C2- C , alky (Co - C , alkyl )C (O )OC2 - C18 alkynyl, (Co -C , alkyl) C (O )OH , nyl, heteroalkyl, (Co - C , alkyl) aryl , (Co - C , alkyl) heteroaryl , ( Co- C , alkyl ) C ( O ) O aryl, (Co - C , alkyl) C ( O ) O heteroaryl, (Co - Cg alkyl ) C ( O ) C - C , alkyl, (Co - Cz alkyl ) C ( O )C2 - Cg alk ( Co- Co alkyl) OC (O ) C - C18 alkyl, (Co - C , alkyl) OC ( O )Cz - 5 enyl, (Co - Cg alkyl) C ( O )C2 - Cz alkynyi , (Co - C , alkyl) C (OH , C18 alkenyl , (Co - C , alkyl) OC ( O )C2 - C18 alkynyl, (Co - Cg ( Co- Cg alkyl) C ( O )aryl , (Co - C , alkyl ) C ( O ) heteroaryl , (Co - C , alkyl) C ( O )NR240 , - C18 alkyl, (Co - C , alkyl) C (O )NR24C2 - alkyl) C (O )OC , -C , alkyl, (Co - C , alkyl) C (O )OC2 - Cg alkenyl, Cig alkenyl, ( C . - C . alkyl ) C ( O )NR24C2 - C18 alkynyl, ( C . - C . (Co - C , alkyl ) C ( O )OC2 - C3 alkynyl, (Co -Cg alkyl) C ( O )OH , alkyl) C ( O )NR24H ,, (Co -C , alkyl) C (O )NR24aryl , (Co - C 0 Co -Cg alkyl) C ( O ) O aryl, (Co - C , alkyl ) C ( O ) O heteroaryl , alkyl) C ( O )NR24heteroaryl , ( C . - C , alkyl) NR24C ( O ) C , - C18 . (Co - C , alkyl) C ( O )NR24C , - C , alkyl, (Co - C , alkyl ) C ( 0 ) alkyl, (Co - C , alkyl) NR24C ( O )C2 - C18 alkenyl, or (Co -Co - NR24C2- C , alkenyl, ( C . - C , alkyl) C ( O ) NR24C2- C , alkynyl, alkyl) NR24C ( O )C2 -C18 alkynyl, (Co - C , alkyl) NR24C ( O ) ( C . - C , alkyl) C ( O ) NR24H2, (Co - C , alkyl) C ( O )NR24aryl , or OH , (Co - C , alkyl) OC (O )OC , -C18 alkyl, ( Co - C , alkyl) OC (Co - C , alkyl) C ( O )NR24heteroaryl ; and ( O )OCZ - C18 alkenyl, (Co - C , alkyl )OC (O )OC2 - C18 alkynyl, (Co - C8 alkyl )OC ( O )OH , (C . - C , alkyl )OC ( O )NR240 , -C18 R24 is hydrogen or C , - C , alkyl. alkyl, (Co - C , alkyl) OC ( O )NR24C2 - C18 alkenyl, ( Co- C , 15 For example , R ' is hydrogen or absent; R² is hydrogen or alkyl) OC ( O )NR24C2 - C18 alkynyl, (Co - C , alkyl) OC ( O ) methyl; R * is hydrogen or methyl ; and Rº is H or absent. NR24H2, (C . -C alkyl) NR24 (O )OC ,- Cis alkyl , ( C .- C , alkyl) Nonlimiting examples of compounds of Formula F NR24 (0 )0C2 - C18 alkenyl , (Co - C , alkyl)NR24 ( O )OC2 - C18 include : alkynyl , or (Co - C , alkyl )NR24 (O )OH ; R is hydrogen , (Co - C , alkyl) halo , C . - C18 alkyl , C2 -C18 20 alkenyl, C2- C18 alkynyl , heteroalkyl, (C . - C alkyl) aryl , (Co Me OH , Cg alkyl) heteroaryl , (C .- C , alkyl) OC , -C18 alkyl , (Co - C , alkyl) OCZ - C18 alkenyl, (Co - Cz alkyl) OC2 - C18 alkynyl, ( Co Me Cg alkyl) OH , (Co - Cg alkyl) SH , (Co - C , alkyl) NR24C7 -C18 alkyl, ( C . - C , alkyl) NR24C2 - C18 alkenyl , (Co - C , alkyl ) 25 NR24C2 -C18 alkynyl, ( Co- Cg alkyl) NR24H2 , ( Co - C , alkyl) C ( O ) C , -C18 alkyl, ( Co- C , alkyl) C ( O )C2 - C18 alkenyl, ( Co- Cg alkyl) C ( O )C2 - C18 alkynyl, (Co - C , alkyl) C ( O ) H , (Co - C , alkyl ) C ( O )aryl , (Co - C , alkyl ) C ( O ) heteroaryl , (Co - Cg alkyl) Testosterone C ( O )OC , -C18 alkyl, (Co - C , alkyl) C ( O ) OC2- C18 alkenyl, 30 Me , (Co - C , alkyl )C (O )OC2 - C18 alkynyl, (Co -C , alkyl) C (O )OH , (Co - C , alkyl) C ( O ) O aryl, (Co - C , alkyl) C ( O ) O heteroaryl, ( Co- C , alkyl) OC (O )C ,- C18 alkyl, (Co -C , alkyl) OC ( O )C2 Me C18 alkenyl, (Co -C , alkyl) OC (O )C2 - C18 alkynyl, (Co -C , alkyl) C ( O )NR24C ,- C18 alkyl, (Co -C , alkyl) C ( O )NR24C2 - 35 H C18 alkenyl, ( Co- C , alkyl) C ( O )NR24C2 -C18 alkynyl, (Co - Cg alkyl) C ( O )NR24H2 , (Co -C , alkyl) C ( O )NR24aryl , (Co -C , alkyl )C ( O )NR24heteroaryl , (Co - C , alkyl )NR24C ( O ) C , -C18 Dehydroepiandrosterone

alkyl, ( Co- C , alkyl) NR24C ( 0 )C2 - C18 alkenyl, or ( Co- C80 Me alkyl) NR24C ( O )C2 - C18 alkynyl, (Co - C , alkyl) NR24C ( O ) 40 OH , (Co - C , alkyl) OC ( O )OC , -C18 alkyl, (Co - C , alkyl) OC ( O )OC2 - C18 alkenyl, (Co - Cg alkyl )OC (O ) OC2- C18 alkynyl , Me (Co - C , alkyl) OC (O )OH , (Co -C , alkyl) OC (O )NR24C , -C18 alkyl, (Co - C , alkyl) OC (O )NR24C2 - C18 alkenyl, (Co - C , alkyl) OC ( O )NR24C2 - C18 alkynyl, (Co - C , alkyl) OC ( O ) 45 NR24H2, (Co -Cz alkyl )NR24 (O )OC - Cig alkyl, (Co - Cz alkyl ) NR24 (O )OC2 - C18 alkenyl, (Co - C , alkyl) NR24 (O )OCZ - C18 Androstenedione alkynyl, or (Co -Cg alkyl) NR24 (O )OH ; R® is hydrogen , C .- C18 alkyl, C2 -C18 alkenyl, C2- C18 Me OH , alkynyl, heteroalkyl, (Co -C , alkyl) aryl, (Co -C , alkyl) het - 50 eroaryl , (Co -Cg alkyl) C ( 0 ) C , -C18 alkyl, (Co - Cg alkyl) C ( O ) Me C2- C18 alkenyl, (Co - Cg alkyl) C (O )C2 - C18 alkynyl, (Co - Cg alkyl) C ( O )H , (Co - Cg alkyl) C (O )aryl , (C .- C . alkyl ) C (O )het eroaryl, (Co - C , alkyl) C ( O )OC -C18 alkyl, (Co - C , alkyl) C ( O )OC2 - C18 alkenyl, (Co - C , alkyl) C ( O )OC2 - C18 alkynyl, 55 HO ( Co- C , alkyl) C ( O )OH , C . - C , alkyl) C ( O ) O aryl, (Co - Cg alkyl) C ( 0 ) 0 heteroaryl, (Co - C , alkyl ) C ( O )NR24C7 - C18 5 - Androstenediol alkyl, (Co -Cg alkyl) C ( O )NR24C2 - C18 alkenyl, ( Co -Cg alkyl ) Me , C ( O )NR24C2 - C18 alkynyl, (Co - C , alkyl) C ( O )NR24H2 , (Co Cg alkyl) C ( O ) NR24aryl, ( C . - C alkyl) C ( O )NR24heteroaryl , 60 Me or SOZH ; and I R24 is hydrogen or C , - C18 alkyl. In some embodiments , Y comprises a structure of For mula E , wherein R ! is hydrogen , C . - C , alkyl; ( Co- Cz alkyl) C ( O ) C7 - C , 65 alkyl, (Co -Cz alkyl) C ( O ) aryl, or SOZH ; Androsterone R² is hydrogen , halo , OH , or CZ - C , alkyl; US 9 ,783 ,592 B2 45 46 - continued In embodiments wherein Y comprises a structure that Me OH , permits or promotes agonist or antagonist activity at a TR , Y is conjugated to L ( e. g . when L is a linking group ) or Q ( e . g . when L is a bond ) at any position of Y that is capable Me of reacting with Q or L . One skilled in the art could readily determine the position of conjugation on Y and means of conjugation of Y to Q or L in view of general knowledge and 11111 the disclosure provided herein . In some embodiments , Y is conjugated to L or Q through any position of Y . In some embodiments , Y is conjugated to L or Q through the car Dihydrotestosterone 10 boxylic acid or alcohol moieties, as indicated below : and derivatives thereof. In embodiments wherein Y comprises a structure of Formula F , Y is conjugated to L ( e . g . when L is a linking group ) or Q ( e . g . when L is a bond ) at any position of 15 Formula F that is capable of reacting with Q or L . One skilled in the art could readily determine the position of H2N conjugation on Formula F and means of conjugation of Formula F to Q or L in view of general knowledge and the disclosure provided herein . In some embodiments, Formula F is conjugated to L or Q at any of positions 1 , 2 , 3 , 4 , 5 , 6 , 20 7 , 8 , 9 , 10 , 11 , 12 , 13 , 14 , 15 , 16 , 17 , 18 , 19 , 20 , 21 , or 22 of Formula F . In some embodiments , Formula F is conju Thyroxine (T4 ) gated to L or Q at position 3 or 17 of Formula F . In some embodiments , the binding of the NHR ligand to the Type I nuclear hormone receptor results in agonist 25 In some embodiments , Y acts at a retinoic acid receptor activity (or antagonist activity ) in some but not all cells or (e .g . RARA , RARB , RARY) . In some embodiments , Y tissues expressing the Type I nuclear hormone receptor. comprises any structure that permits or promotes agonist NHR Ligand that Acts on a Type II Nuclear Hormone activity at the RAR , while in other embodiments Y is an Receptor antagonist of RAR . In exemplary embodiments , Y com In some embodiments of the invention , the NHR ligand 30 prises a structure of Formula G : ( Y ) acts on a Type II nuclear hormone receptor . In some embodiments , Y can have any structure that permits or promotes agonist activity upon binding of the ligand to a Formula G Type II nuclear hormone receptor, while in other embodi CH3 ments Y is an antagonist of the Type II nuclear hormone HzCH?C CH2CH? CH3 receptor. In exemplary embodiments , Y exhibits agonist ( or 35 RII antagonist) activity at a thyroid hormone receptor ( TR ), retinoic acid receptor (RAR ) , peroxisome proliferator acti vated receptor ( PPAR ) , Liver X Receptor (LXR ) , farnesoid X receptor (FXR ) , vitamin D receptor (VDR ), and /or preg CH3 nane X receptor (PXR ) . 40 In some embodiments , Y acts at a thyroid hormone wherein Rl is a moiety that permits or promotes agonist or receptor ( e . g . TRA , TRB ) . In some embodiments , Y com - antagonist activity upon the binding of the compound of prises any structure that permits or promotes agonist activity Formula G to a RAR , and www represents either E or Z at the TR , while in other embodiments Y is an antagonist of stereochemistry . TR . Nonlimiting examples of Y includede the following com - 4516 In some embodiments , Y comprises a structure of For pounds: mula G wherein R11 is C ( O ) OH , CH ,OH , or C (O ) H . In some embodiments , Y comprises a structure of Formula G wherein R11 is a carboxylic acid derivative ( e . g . acyl chlo ride , anhydride , and ester ) . Nonlimiting examples of the compound of Formula G HNH?N — E 50 include: OH , H3C CH3 CH3 CH3 0

OH , Thyroxine ( T4) 55 V OH All - trans - retinoic acid HN 60 H3C CH3 CH3 CH3 - OH ,

CH3 Triiodothyroxine ( T3 ) 65 tool trianRetinol and derivatives thereof. US 9 ,783 ,592 B2 48 -continued arachidonic acid , dihydroxyeicosatetraenoic acid (Di H3C CH3 CH3 CH3 0 HETE ), octadecynoic acid , eicosatriynoic acid , eicosadi enoic acid , eicosatrienoic acid , eicosapentaenoic acid , erucic acid , dihomolinolenic acid , docosatrienoic acid , docosapen 5 taenoic acid , docosahexaenoic acid , adrenic acid , and CH3 derivatives thereof. Retinal CH3 Hzc CHZ 10 F2c F F F F F F C V CHCH3 ; HzCH3C Perfluorononanoic acid F F F F F F 0 OH , F3CX OH 11 - cis -retinoic acid F FF FF F? 20 and derivatives thereof. Perfluorooctanoic acid In embodiments wherein Y comprises a structure of Formula G , Y is conjugated to L ( e . g . when L is a linking In embodiments wherein Y comprises a structure of group ) or Q (e . g . when L is a bond ) at any position of Formula H , Y is conjugated to L ( e. g. when L is a linking Formula G that is capable of reacting with Q or L . One 25 group ) or Q (e . g . when L is a bond ) at any position of skilled in the art could readily determine the position of Formula H that is capable of reacting with Q or L . One conjugation on Y and means of conjugation of Y to Q or L skilled in the art could readily determine the position of in view of general knowledge and the disclosure provided conjugation on Formula H and means of conjugation of herein . In some embodiments , Y is conjugated to L or Q Formula H to Q or L in view of general knowledge and the through any position of Y . In some embodiments , Formula 30 disclosure provided herein . In some embodiments , Formula G is conjugated to L or Q at R11 . H is conjugated to L or Q at any position on Formula H . In In some embodiments , Y acts at a peroxisome proliferator some embodiments , Formula H is conjugated to L or activated receptor (e . g . PPARA , PPARB / 8 , PPARY) . In some mythrough the terminal carboxylic acid moiety . embodiments , Y comprises any structure that permits or In some of these embodiments , Y is an eiconsanoid . In promotes agonist activity at the PPAR , while in other 35 specific embodiments , Y is a prostaglandin or a leukotriene . embodiments Y is an antagonist of PPAR . In some embodi- In some exemplary embodiments , Y is a prostaglandin ments , Y is a saturated or unsaturated , halogenated or having a structure as described by Formulae J1 - J6 : nonhalogenated free fatty acid (FFA ) as described by For mula H : 40 Formula J1 Formula H - R13 R 12 (R12 HO R12 YR 4545 R 13 TR12 Formula J2 ö R 12 ?? wherein n is 0 - 26 and each R12 , when present, is indepen dently a moiety that permits or promotes agonist or antago - 50 nist activity upon binding of the compound of Formula H to a PPAR . In some embodiments , Y comprises a structure of For Formula J3 mula H , wherein n is 0 - 26 and each R12 , when present , is independently hydrogen , C . - C , alkyl , or halogen . In some 55 embodiments Formula B is saturated such as , for example , formic acid , acetic acid , n -caproic acid , heptanoic acid , caprylic acid , nonanoic acid , capric acid , undecanoic acid , R 13 lauric acid , tridecanoic acid , myristic acid , pentadeconoic HO acid , palmitic acid , heptadecanoic acid , stearic acid , non - 60 Formual J4 adecanoic acid , arachidic acid , heneicosanoic acid , behenic HO acid , tricosanoic acid , perfluorononanoic acid ( see below ) , - R13 perfluorooctanoic acid (see below ), and derivatives thereof. In some embodiments Formula H is unsaturated with either cis or trans stereochemistry such as , for example , mead acid , 65 R13 myristoleic acid , palmitoleic acid , sapienic acid , oleic acid , linoleic acid , a - linolenic acid , elaidic acid , petroselinic acid , US 9 ,783 , 592 B2 49 50 - continued Formula J5 OH OH R13 OH . R 13 R13 Formula J6 In some embodiments when Y comprises a structure of 10 Formula K , each R14 is independently C3- C13 alkyl, C3- C13 alkenyl, C3 -C13 alkynyl, or heteroalkyl. In embodiments wherein Y comprises a structure of Formula K , Y is conjugated to L ( e . g . when L is a linking group ) or Q ( e . g . when L is a bond ) at any position of R 13 15 Formula K that is capable of reacting with Q or L . One skilled in the art could readily determine the position of conjugation on Formula K and means of conjugation of wherein each R13 is independently a moiety that permits or Formula K to Q or L in view of general knowledge and the promotes agonist or antagonist activity upon the binding of disclosure provided herein . In some embodiments , Formula the compound of Formula J to a PPAR ( e . g . PGJ2 as shown 20 K is conjugated to L or Q at any position on Formula K . In below ) : some embodiments , Formula K is conjugated to L or Q through the terminal carboxylic acid moiety or through a pendant alcohol moiety . In some exemplary embodiments , Y is a thiazolidinedione 25 comprising a structure as described by Formula L : 1111111

Formula L

30 HOI11.0 NH .

35 CH3 Nonlimiting examples of the compound of Formula L In some embodiments when Y comprises a structure of include : any one of Formulae J1 - J6 , each R13 is independently CZ- C , alkyl, C . - C , alkenyl, C7- C , alkynyl, or heteroalkyl. In embodiments wherein Y is an eicosanoid , Y is conju gated to L ( e. g . when L is a linking group ) or Q ( e. g . when L is a bond ) at any position of the eicosanoid that is capable NH , of reacting with Q or L . One skilled in the art could readily 45 determine the position of conjugation on and means of conjugation of Y to Q or L in view of general knowledge and the disclosure provided herein . In some embodiments , Y is Rosiglitazone conjugated to L or Q through any position of Y . In some embodiments , the eicosanoid is conjugated to L or Q 50 through a terminal carboxylic acid moiety or through a pendant alcohol moiety . In some exemplary embodiments, Y is a leukotriene having a structure as described by Formula K or a deriva - H3C tized form of Formula K : 55 Pioglitazone Formula K H3CH3C . oras LR14 60 - R14 HOM

HC wherein each R14 is independently a moiety that permits or promotes agonist or antagonist activity upon the binding of 65 Troglitazone the compound of Formula K to a PPAR ( e . g . leukotriene B4 Brane as shown below ) : and derivatives thereof. US 9 ,783 ,592 B2 51 In embodiments wherein Y comprises a structure of In some embodiments , Y acts at a liver X receptor (LXRa , Formula L , Y is conjugated to L ( e . g . when L is a linking LXRB ) . In some embodiments , Y comprises any structure group ) or Q ( e . g . when L is a bond ) at any position of that permits or promotes agonist activity at the LXR , while Formula L that is capable of reacting with Q or L . One in other embodiments Y is an antagonist of LXR . In exem skilled in the art could readily determine the position of 5 plary embodiments , Y is an oxysterol (i . e . oxygenated conjugation on Formula L and means of conjugation of derivative of cholesterol) . Nonlimiting examples of Y in Formula L to Q or L in view of general knowledge and the these embodiments include 22 (R )- hydroxycholesterol (see disclosure provided herein . In some embodiments , Formula below ) , 24 ( S ) -hydroxycholesterol (see below ) , 27 -hydroxy L is conjugated to L or Q at any position on Formula L , such cholesterol, cholestenoic acid , and derivatives thereof . as, for example , a pendant alcohol moiety , or through an OH aromatic substituent. In some embodiments , Y acts at a RAR - related orphan Meln. Me receptor (e . g. RORA , RORB , RORY) . In some embodiments , Me Y comprises any structure that permits or promotes agonist 16 activity at the ROR , while in other embodiments Y is an Me Me antagonist of ROR . > Nonlimiting examples of Y include : LI 20 HO @nin 22 (R ) -Hydroxycholesterol OH Me Me Meni Me Me . Me Me .)

HO I IM Cholesterol 30 HO CH3, 24 ( S ) - Hydroxycholesterol -- HN In embodiments wherein Y acts at a LXR , Y is conjugated to L ( e . g . when L is a linking group ) or Q ( e . g . when L is a 35 bond ) at any position of Y that is capable of reacting with Q or L . One skilled in the art could readily determine the Han position of conjugation on Y and means of conjugation of Y to Q or L in view of general knowledge and the disclosure provided herein . In some embodiments , Y is conjugated to 40 L or Q at any of positions 1 , 2 , 3, 4 , 5 , 6 , 7 , 8 , 9 , 10 , 11, 12 , Melatonin 13 , 14 , 15 , 16 , 17 , 18 , 19 , 20 , 21 , 22, 23 , 24 , 25 , or 26 of Formula F . In some embodiments , Formula F is conjugated HC' to L or Q at position 3 or 17 of Formula F . CH3, In some embodiments , Y acts at the farnesoid X receptor NH NH 45 (FXR ). In some embodiments , Y comprises any structure that permits or promotes agonist activity at the FXR , while CGP 52608 in other embodiments Y is an antagonist of FXR . In some of CH , 0 these embodiments , Y is a bile acid . In exemplary embodi HzC CH3 CH3 ments , Y has a structure of Formula M : 50 " OH , Formula M

Me All- trans- retinoic acid 55 III. RI A Me and derivatives thereof . In embodiments wherein Y acts at a ROR , Y is conjugated to L ( e . g. when L is a linking group ) or Q ( e . g . when L is a 60 bond ) at any position of Y that is capable of reacting with Q or L . One skilled in the art could readily determine the H position of conjugation on Y and means of conjugation of Y wherein each of R15 , R1 , and R17 are independently moi to Q or L in view of general knowledge and the disclosure eties that permit or promote agonist or antagonist activity provided herein . In some embodiments , Y is conjugated to 65 upon binding of the compound of Formula M to a FXR . L or Q through any position of Y , such as , for example , any In some embodiments when Y comprises a structure of of the positions previously described herein . Formula M , each of R15 and R16 are independently hydro US 9 ,783 , 592 B2 53 54 gen , (Co - Cg alkyl) halo , C . -C1g alkyl, C2 -C1g alkenyl, C2- C18 - continued alkynyl, heteroalkyl, or ( Co- C , alkyl) OH ; and R17 is OH , (Co -C , alkyl) NH (C ,- C4 alkyl )SO3H , or (Co - C , alkyl )NH OH ( C - C4 alkyl) COOH . 11. Me In some embodiments when Y comprises a structure of 5 NH Formula M , each of R15 and R16 are independently hydrogen IH or OH ; and R17 is OH , NH (C , -C2 alkyl )SO3H , or NH (C , -C2 IM) alkyl) COOH . Nonlimiting examples of the compound of Formula M include : M 1010 HOILUHO V " OH Mer Glycocholic acid OH Me OH , 15 and derivatives thereof. Me In embodiments wherein Y comprises a structure of - Formula M , Y is conjugated to L (e .g . when L is a linking ?H ** group ) or Q ( e . g . when L is a bond ) at any position of OH 20 Formula M that is capable of reacting with Q or L . One skilled in the art could readily determine the position of Cholic acid conjugation on Formula M and means of conjugation of Men Formula M to Q or L in view of general knowledge and the Me 25 disclosure provided herein . In some embodiments , Formula OH . M is conjugated to L or Q at any of positions 1 , 2 , 3 , 4 , 5 , Me 6 , 7 , 8 , 9 , 10 , 11 , 12 , 13 , 14 , 15 , 16 , 17 , 18 , 19 , 20 , 21, 22 , 23 , 24 , or 25 of Formula M . In some embodiments , Formula 30 M is conjugated to L or Q at position 3 , 7 , 12 or 17 of Formula M . In some embodiments , Y acts at the vitamin D receptor Deoxycholic acid (VDR ). In some embodiments, Y comprises any structure Me, 35 that permits or promotes agonist activity at the VDR , while Me in other embodiments Y is an antagonist of VDR . In exem plary embodiments , Y has a structure of Formula N : Me

Formula N R22 R21 LowR23 PartyLithocholic acid Me, Me OH , H

* HOM11 H R 18 Chenodeoxycholic acid Me -Pose OH wherein each of R18 , R19 , R20 , R21, R22 , and R23 are En.E Me O= moieties that permit or promote agonist or antagonist activ IZ ity upon binding of the compound of Formula N to the VDR such as, for example , any of the vitamin D compounds found in Bouillon et al ., Endocrine Reviews, 16 ( 2 ): 200 - 257 ( 1995 ) . ?H In some embodiments wherein Y comprises a structure of HO11111 “?? Formula N , 65 Taurocholic acid R18 and Rl' are each independently hydrogen , (Co - C , alkyl) halo , (Co -Cg alkyl) heteroaryl , or (Co - Cg alkyl) OH ; US 9 , 783 ,592 B2 55 56 both of R20 are hydrogen or both of R20 are taken together -continued to form = CH2; Me each of R21 and R22 are independently C ,- C4 alkyl; and Me R23 is C4 -C18 alkyl, C4- C1g alkenyl, C4- C1g alkynyl, het 5 eroalkyl , ( C4- C18 alkyl) aryl , ( C4 -C18 alkyl) heteroaryl, ( Co — Me, Cg alkyl )OC - C18 alkyl, ( Co- C , alkenyl )OC -C18 alkyl, (Co Me OH C , alkynyl) OC ,- C18 alkyl, (C .- C , alkyl) OC2 - C18 alkenyl, (Co - C , alkyl) OC2 - C18 alkynyl, ( C . -C18 alkyl) OH , (Co -C18 10 alkyl) SH , (C6 - C1g alkenyl) OH , ( Co- C18 alkynyl) OH , (Co - C , alkyl) NR24C7 -C18 alkyl, (Co - C , alkenyl) NR24C7 -C18 alkyl , (Co - C , alkynyl) NR240 , -C18 alkyl, (Co - C , alkyl )NR24C2 C18 alkenyl, ( Co -Cg alkyl )NR24C2 - C18 alkynyl, (Co - Cg 15 alkyl) C ( O )C1 - C18 alkyl, (Co -Cg alkyl ) C ( O )C2 -C18 alkenyl, 25- Hydroxyvitamin D3 (Co - C , alkyl) C ( O ) C2- C18 alkynyl, (Co -C , alkyl) C (O ) H , (Co and derivatives thereof. In embodiments wherein Y comprises a structure of Cg alkyl) C ( O )aryl , ( Co - C , alkyl ) C (O )heteroaryl , (Co - Cg 20 Formula N , Y is conjugated to L ( e . g . when L is a linking alkyl ) C ( O )OC7 - C18 alkyl, (Co - C , alky1) C ( O )OC2 - C18 alk group ) or Q ( e . g . when L is a bond ) at any position of enyl, (Co -C , alkyl) C (O )OC2 - C18 alkynyl, ( Co - Cg alkyl) C (O ) Formula N that is capable of reacting with Q or L . One OH , ( Co - C , alkyl ) C ( O )O aryl, ( Co- Cg alkyl) C ( O ) O het skilled in the art could readily determine the position of eroaryl, (Co - Cg alkyl )OC ( O ) C -C18 alkyl, (Co - C , alkyl )OC conjugation on Formula N and means of conjugation of ( 0 )C2 - C18 alkenyl, (Co - C , alkyl) OC ( O )C2 - C18 alkynyl, (Co - 25 Formuladisclosure N providedto Q or L hereinin view . In ofsome general embodiments knowledge , Formulaand the Cg alkyl) C (O )NR24C7 -C18 alkyl, (C .- C alkyl )C ( 0 ) N is conjugated to L or at any of positions 1, 2, 3 , 4 , 5 , 6 , 7 , 8 , 9 , 10 , 11 , 12 , 13 , 14 , 15 , 16 , 17 , 18 , 19 , 20 , 21, 22 , NR24C2 -C18 alkenyl, ( Co- Co alkyl) C (O )NR24C2 - C18 23 , 24 , 25 , or 26 of Formula N . In some embodiments , alkynyl, (Co -Co alkyl ) C ( O )NR H2, ( Co -Co alkyl) C ( O ) - Formula N is conjugated to L or O at position 1 . 3 . 19 . or 25 NR24aryl, (Co - C , alkyl ) C (O )NR24heteroaryl , (Co - C , alkyl) of Formula N . In some embodiments , Y acts at the pregnane X receptor NR24C ( O ) C , -C18 alkyl , ( Co- C , alkyl) NR24C ( O )C2 - C18 alk (PXR ). In some embodiments , Y comprises any structure enyl, or (Co - Cg alkyl) NR24C ( O )C2 - C18 alkynyl , ( Co- Cg that permits or promotes agonist activity at the PXR , while alkyl )NR24C ( O )OH , ( Co- Co alkyl) OC ( O ) OC , - C18 alkyl , in other embodiments Y is an antagonist of PXR . In some embodiments , Y is a steroid , antibiotic , antimycotic , bile (Co - Cg alkyl) OC ( O )OC2 - C18 alkenyl, (Co - C , alkyl) OC ( O ) acid , hyperforin , or a herbal compound . In exemplary OC2- C18 alkynyl , ( Co - C , alkyl) OC ( O )OH , (Co - C , alkyl) OC embodiments , Y is compound that is able to induce CY3A4 , (O )NR24C7 -C18 alkyl, (Co -C , alkyl) OC (O )NR24C2 - C18 alk such as dexamethasone and rifampicin . In embodiments wherein Y comprises a structure that acts at the PXR , Y is enyl, (Co - C , alkyl )OC ( O )NR24C2 - C18 alkynyl, (Co -Cg 40 conjugated to L ( e . g . when L is a linking group ) or Q ( e . g . alkyl) OC ( O )NR24H2 , (Co - C , alkyl) NR24 ( O ) OC , - C18 alkyl , when L is a bond ) at any position of Y that is capable of ( Co- C , alkyl) NR24 (0 ) OC2- C18 alkenyl, ( Co - C , alkyl) NR24 reacting with Q or L . One skilled in the art could readily determine the position of conjugation on Y and means of ( O )OC2 - C18 alkynyl, or (Co - C , alkyl) NR24 ( 0 )OH ; and conjugation of Y to Q or L in view of generalknowledge and R24 is hydrogen or C7- C18 alkyl. the disclosure provided herein . In some embodiments , Y is conjugated to L or Q at any of positions on Y . Nonlimiting examples of the compound of Formula N Modification of the NHR Ligand ( Y ) include : In some embodiments , the NHR ligand is derivatized or otherwise chemically modified to comprise a reactive moi ety that is capable of reacting with the glucagon superfamily 50 peptide ( Q ) or the linking group ( L ) . In the embodiments described herein , Y is derivatized at any position of Y that is capable of reacting with Q or L . The position of deriva tization on Y is apparent to one skilled in the art and depends on the type of NHR ligand used and the activity that is 55 desired . For example , in embodiments wherein Y has a Me , structure comprising a tetracyclic skeleton having three .11 Me OH 6 -membered rings joined to one 5 -membered ring or a variation thereof, Y can be derivatized at any of positions 1, 2 , 3 , 4 , 5 , 6 , 7 , 8 , 9 , 10 , 11, 12 , 13 , 14 , 15 , 16 , 17 , 18, 19 , 20 , 21, 22 , 23 , 24 , or 25 . Other positions of derivatization can be as previously described herein . The NHR ligand can be derivatized using any agent known to one skilled in the art or described herein (e . g . see The Linking Group section and the Chemical Modification V OH of Q and / or Y subsection ) . For example , estradiol can be 65 derivatized with succinic acid , succinic anhydride, benzoic Calcitriol acid , ethyl 2 - bromoacetate , or iodoacetic acid to form the below derivatives of estradiol US 9 ,783 , 592 B2 57 58

Me OH

HO succinic acid benzoic acid ethyl 2 -bromoacetate or or succinic anhydride iodoacetic acid Me OH Me OH Me OH

I HO . HO IZ dipolor or

mecresMe MO Me OH poprohouse

HO HO ' ?? comoSimilarly , any of the aforementioned NHR ligands can be » PACAP - 27 , native PHM , native oxyntomodulin , native derivatized by methods known in the art. Additionally , secretin , or native GIP with up to 1 , 2 , 3 , 4 , 5 , 6 , 7 , 8 , 9 or certain derivatized ligands are commercially available and 10 amino acid modifications. can be purchased from chemical companies such as Sigma In some aspects of this invention , Q is a glucagon related Aldrich . 40 peptide such as , for example , glucagon (SEQ ID NO : 1601 ), The Glucagon Superfamily Peptide ( Q ) oxyntomodulin ( SEQ ID NO : 1606 ) , exendin - 4 ( SEQ ID In the Q -L - Y conjugates described herein , Q is a glucagon NO : 1618 ) , glucagon - like peptide - 1 (GLP - 1 ) (amino acids superfamily peptide . A glucagon superfamily peptide refers 7 - 36 provided as SEQ ID NO : 1603 ; amino acids 7 - 37 are to a group of peptides related in structure in their N - terminal provided as SEQ ID NO : 1604 ) , glucagon - like peptide - 2 and / or C - terminal regions ( see , for example , Sherwood et 45 (GLP - 2 , SEQ ID NO : 1608 ) , gastric inhibitory peptide (GIP , al. , Endocrine Reviews 21 : 619 -670 ( 2000 ) . It is believed SEQ ID NO : 1607 ) or analogs, derivatives and conjugates that the C - terminus generally functions in receptor binding thereof . A glucagon related peptide has biological activity and the N - terminus generally functions in receptor signal- (as an agonist or antagonist ) at any one or more of the ing . A few amino acids in the N - terminal and C - terminal glucagon , GLP - 1 , GLP - 2 , and GIP receptors and comprise regions are highly conserved among members of the gluca - 50 an amino acid sequence that shares at least 20 % sequence gon superfamily . Some of these conserved amino acids identity ( e . g . , 25 % , 30 % , 35 % , 40 % , 45 % , 50 % , 55 % , 60 % , include His1 , Gly4 , Pheh , Phe22 , Val23 , Trp25 and Leu26 , 65 % , 70 % , 75 % , 80 % , 85 % , 90 % , 95 % ) with at least one of with amino acids at these positions showing identity , con native glucagon , native oxyntomodulin , native exendin - 4 , native GLP - 1 ( 7 - 37 ) , native GLP - 2 , or native GIP over the servative substitutions or similarity in the structure of their 55 length of the peptide (or over the positions which correspond amino acid side chains. to glucagon , see e . g . FIG . 1 ). Glucagon superfamily peptides include glucagon related It is understood that all possible activity subsets of peptides , Growth Hormone Releasing Hormone (GHRH ; glucagon related peptides are contemplated , e. g . peptides SEQ ID NO : 1619 ), vasoactive intestinal peptide (VIP ; SEQ which have biological activity (as agonists or antagonists ) at ID NO : 1620 ) , pituitary adenylate cyclase - activatingating poly - 60 any one or more of the glucagon or GLP - 1 or GIP receptors , peptide 27 (PACAP - 27 ; SEQ ID NO : 1621) , peptide histi together with all possible subsets of sequence identity to dine isoleucine (PHI ; SEQ ID NO : 1542) , peptide histidine each listed native peptide , e . g . , comprise an amino acid methionine (PHM ; SED ID NO : 1622 ), secretin (SEQ ID sequence that shares at least 20 % , 25 % , 30 % , 35 % , 40 % , NO : 1623 ), and/ or analogs, derivatives or conjugates 45 % , 50 % , 55 % , 60 % , 65 % , 70 % , 75 % , 80 % , 85 % , 90 % , or thereof. In some embodiments , Q comprises an amino acid 65 95 % sequence identity with native GLP -1 over the length of sequence of native glucagon , native exendin - 4 , native GLP- native GLP - 1 . In some embodiments of the invention , the 1 (7 -37 ), native GLP -2 , native GHRH , native VIP , native glucagon related peptide is a peptide having glucagon recep US 9 , 783 ,592 B2 59 60 tor agonist activity , GLP - 1 receptor agonist activity , GIP In some embodiments , Q exhibits an EC50 or IC50 at the receptor agonist activity , glucagon receptor/ GLP - 1 receptor glucagon receptor that is about 0 .001 PM or more, about co -agonist activity , glucagon receptor /GIP receptor co -ago - 0 .01 PM or more, or about 0 . 1 PM or more . Glucagon nist activity , GLP - 1 receptor/ GIP receptor co - agonist activ - receptor activation ( glucagon receptor activity ) can be mea ity , glucagon receptor/ GLP - 1 receptor/ GIP receptor tri- ago - 5 sured by in vitro assays measuring cAMP induction in nist activity , glucagon receptor antagonist activity , or HEK293 cells over -expressing the glucagon receptor , e . g. , glucagon receptor antagonist/ GLP - 1 receptor agonist activ - assaying HEK293 cells co - transfected with DNA encoding ity . In some embodiments , the peptide retains an alpha -helix the glucagon receptor and a luciferase gene linked to CAMP conformation in the C - terminal half of the molecule . In some responsive element as described in Example 2 . embodiments , the peptide retains positions involved in 0 In some embodiments , Q exhibits about 0 . 001 % or more , receptor interaction or signaling , e . g . position 3 of glucagon , about 0 .01 % or more , about 0 . 1 % or more , about 0 . 5 % or or position 7 , 10 , 12 , 13 , 15 or 17 of GLP - 1 ( 7 - 37 ) . Accord - more , about 1 % or more , about 5 % or more , about 10 % or ingly , the glucagon related peptide can be a peptide of Class more , about 20 % or more , about 30 % or more , about 40 % 1 , Class 2 , Class 3 , Class 4 , and / or Class 5 , each of which 16 or more , about 50 % or more, about 60 % or more , about 75 % is further described herein . or more , about 100 % or more , about 125 % or more , about Q may also be any of the glucagon superfamily peptides 150 % or more , about 175 % or more , about 200 % or more , that are known in the art, some of which are disclosed herein about 250 % or more, about 300 % or more, about 350 % or by way of nonlimiting examples. A variety ofGLP - 1 analogs more, about 400 % or more, about 450 % or more , or about are known in the art and are a glucagon - related peptide 20 500 % or higher activity at the glucagon receptor relative to according to the current invention , see, e . g ., WO native glucagon ( glucagon potency ) . In some embodiments , 2008023050 , WO 2007030519 , WO 2005058954 , WO Q exhibits about 5000 % or less or about 10 , 000 % or less 2003011892 , WO 2007046834 , WO 2006134340 , WO activity at the glucagon receptor relative to native glucagon . 2006124529 , WO 2004022004 , WO 2003018516 , WO The activity of Q at a receptor relative to a native ligand of 2007124461 each incorporated herein by reference in its 25 the receptor is calculated as the inverse ratio of ECsos for Q entirety for each of its sequence or formula disclosures of versus the native ligand . GLP - 1 analogs or derivatives . In any of the embodiments, Q In some embodiments, Q exhibits substantial activity can be a glucagon related peptide disclosed in WO 2007 / (potency ) at only the glucagon receptor and little to no 056362 , WO 2008 /086086 , WO 2009 / 155527 , WO 2008 / activity at the GLP - 1 receptor or the GIP receptor . In some 101017 , WO 2009/ 155258 , WO 2009/ 058662 , WO 2009 / 30 embodiments , Q is considered as a “ pure glucagon receptor 058734 , WO 2009 /099763 , WO 2010 /011439 , PCT Patent agonist ” or is not considered as a “ glucagon /GLP - 1 receptor Application No . US09 /68745 , and U . S . Patent Application co - agonist ” or a “ glucagon /GIP receptor co -agonist . ” In No . 61 / 187, 578 each incorporated herein by reference in its some embodiments Q exhibits any of the levels of activity entirety. In certain embodiments , Q is a Class 1 , Class 2 , or potency at the glucagon receptor described herein but has Class 3 , Class 4 , or Class 5 glucagon related peptide as 35 substantially less activity ( potency ) at theGLP - 1 receptor or detailed herein . In any of the embodiments described herein , the GIP receptor. In some embodiments , Q exhibits an EC50 O is any of SEO ID NOs: 1 - 760 , 801 -919 , 1001 - 1275 , at the GLP - 1 receptor which is 100 - fold or greater than the 1301 - 1371, 1401 - 1518 , 1601- 1650 . In some embodiments, EC50 at the glucagon receptor. In some embodiments , Q Q is any of SEQ ID NOs : 1647 - 1650 . exhibits an EC50 at the GIP receptor which is 100 - fold or Activity of the Glucagon Superfamily Peptide ( Q ) 40 greater than the EC50 at the glucagon receptor . Activity at the Glucagon Receptor Activity at the GLP - 1 Receptor In some embodiments , Q exhibits an EC50 for glucagon In some embodiments , Q exhibits an EC50 for GLP -1 receptor activation ( or an IC5o for glucagon receptor antago - receptor activation ( or an IC50 for GLP - 1 receptor antago nism ) of about 10 mM or less , or about 1 mM ( 1000 uM ) or nism ) of about 10 mM or less, or about 1 mM ( 1000 uM ) or less ( e . g . , about 750 uM or less, about 500 uM or less , about 45 less ( e . g . , about 750 uM or less , about 500 uM or less , about 250 UM or less, about 100 uM or less, about 75 uM or less , 250 UM or less , about 100 uM or less , about 75 uM or less , about 50 uM or less , about 25 uM or less , about 10 uM or about 50 uM or less , about 25 uM or less , about 10 uM or less , about 7. 5 uM or less , about 6 uM or less , about 5 uM less, about 7 .5 uM or less, about 6 uM or less, about 5 uM or less , about 4 uM or less , about 3 uM or less , about 2 uM or less , about 4 uM or less , about 3 uM or less, about 2 uM or less or about 1 uM or less ) . In some embodiments , Q 50 or less or about 1 uM or less ) . In some embodiments , Q exhibits an EC 50 or IC5, at the glucagon receptor of about exhibits an EC50 or IC50 for GLP - 1 receptor activation of 1000 nM or less ( e . g ., about 750 nM or less, about 500 nM about 1000 nM or less ( e . g . , about 750 nM or less , about 500 or less , about 250 nM or less, about 100 nM or less, about nM or less, about 250 nM or less , about 100 nM or less , 75 nM or less , about 50 nM or less , about 25 nM or less , about 75 nM or less, about 50 nM or less , about 25 nM or about 10 nM or less , about 7 . 5 nM or less , about 6 nM or 55 less , about 10 nM or less , about 7 . 5 nM or less , about 6 nM less, about 5 nM or less, about 4 nM or less , about 3 nM or or less , about 5 nM or less , about 4 nM or less, about 3 nM less, about 2 nM or less or about 1 nM or less ). In some or less , about 2 nM or less or about 1 nM or less ) . In some embodiments, has an EC50 or IC50 at the glucagon embodiments , Q has an EC50 or IC50 at the GLP - 1 receptor receptor which is in the picomolar range. Accordingly, in which is in the picomolar range . Accordingly, in some some embodiments , Q exhibits an EC50 or IC50 at the 60 embodiments , Q exhibits an EC5o or IC50 for GLP - 1 recep glucagon receptor of about 1000 PM or less ( e . g . , about 750 tor activation of about 1000 PM or less ( e . g . , about 750 PM PM or less, about 500 PM or less , about 250 PM or less , or less , about 500 pM or less, about 250 PM or less, about about 100 PM or less , about 75 PM or less , about 50 PM or 100 PM or less, about 75 PM or less , about 50 PM or less , less, about 25 PM or less , about 10 PM or less , about 7 . 5 PM about 25 PM or less , about 10 PM or less, about 7 . 5 PM or or less, about 6 PM or less, about 5 PM or less , about 4 PM 65 less , about 6 PM or less , about 5 PM or less , about 4 PM or or less , about 3 PM or less, about 2 PM or less or about 1 PM less , about 3 PM or less , about 2 PM or less or about 1 PM or less ) . or less ) . US 9 ,783 , 592 B2 67 62 In some embodiments , Q exhibits an EC 50 or IC50 at the Receptor activation can be measured by in vitro assays GLP - 1 receptor that is about 0 . 001 PM or more , about 0 .01 measuring CAMP induction in HEK293 cells over - express PM or more, or about 0 . 1 PM or more . GLP - 1 receptor ing the GIP receptor, e . g . assaying HEK293 cells co - trans activation (GLP - 1 receptor activity ) can be measured by in fected with DNA encoding the receptor and a luciferase gene vitro assays measuring cAMP induction in HEK293 cells 5 linked to cAMP responsive element as described in Example over -expressing the GLP - 1 receptor, e . g . , assaying HEK293 2 . cells co - transfected with DNA encoding the GLP - 1 receptor In some embodiments of the present disclosures , Q exhib and a luciferase gene linked to cAMP responsive element as its at least or about 0 . 1 % activity of native GIP at the GIP described in Example 2 . receptor. In exemplary embodiments , Q exhibits at least or In some embodiments , Q exhibits about 0 .001 % or more , 10 about 0 . 2 % , at least or about 0 . 3 % , at least or about 0 . 4 % , about 0 .01 % or more , about 0 . 1 % or more , about 0 . 5 % or at least or about 0 . 5 % , at least or about 0 . 6 % , at least or more , about 1 % or more , about 5 % or more , about 10 % or about 0 . 7 % , at least or about 0 . 8 % , at least or about 0 . 9 % , more , about 20 % or more , about 30 % or more , about 40 % at least or about 1 % , at least or about 5 % , at least or about or more, about 50 % or more , about 60 % or more , about 75 % 10 % , at least or about 20 % , at least or about 30 % , at least or more , about 100 % or more , about 125 % or more , about 15 or about 40 % , at least or about 50 % , at least or about 60 % , 150 % or more , about 175 % or more , about 200 % or more , at least or about 70 % , at least or about 75 % , at least or about about 250 % or more , about 300 % or more , about 350 % or 80 % , at least or about 90 % , at least or about 95 % , or at least more , about 400 % or more , about 450 % or more , or about or about 100 % of the activity of native GIP at the GIP 500 % or higher activity at the GLP - 1 receptor relative to receptor. native GLP - 1 (GLP - 1 potency ) . In some embodiments , Q20 In some embodiments of the present disclosures , Q exhib exhibits about 5000 % or less or about 10 , 000 % or less its activity at the GIP receptor which is greater than that of activity at the GLP - 1 receptor relative to native GLP - 1 native GIP . In exemplary embodiments , Q exhibits at least or (GLP - 1 potency ) . about 101 % , at least or about 105 % , at least or about 110 % , In some embodiments, Q exhibit substantial activity (po - at least or about 125 % , at least or about 150 % , at least or tency ) at only the GLP - 1 receptor and little to no activity at 25 about 175 % at least or about 200 % , at least or about 300 % , the glucagon receptor or the GIP receptor. In some embodi- at least or about 400 % , at least or about 500 % or higher % ments , Q is considered as a “ pure GLP - 1 receptor agonist ” of the activity of native GIP at the GIP receptor. In some or is not considered as a “ glucagon /GLP - 1 receptor co - embodiments , Q exhibits no more than 1000 % , 10 , 000 % , agonist” or a “GLP - 1 /GIP co -agonist . ” In some embodi - 100 ,000 % , or 1 , 000 ,000 % activity at the GIP receptor rela ments Q exhibits any of the levels of activity or potency at 30 tive to native GIP . A peptide ' s activity at the GIP receptor the GLP - 1 receptor described herein but have substantially relative to native GIP is calculated as the inverse ratio of less activity ( potency) at the glucagon receptor or the GIP ECsos for the GIP agonist peptide vs . native GIP . receptor. In some embodiments , Q exhibits an EC5, at the In some embodiments , exhibits substantial activity glucagon receptor which is 100 - fold or greater than the EC50 (potency ) at only the GIP receptor and little to no activity at at the GLP - 1 receptor. In some embodiments , Q exhibits an 35 the glucagon receptor or the GLP - 1 receptor. In some EC50 at the GIP receptor which is 100 - fold or greater than embodiments , Q is considered as a " pure GIP receptor the EC 50 at the GLP - 1 receptor. agonist” or is not considered as a “ glucagon /GIP receptor Activity at the GIP Receptor co - agonist ” or a “ GLP - 1 /GIP co -agonist .” In some embodi In some embodiments , Q exhibits an EC50 for GIP recep ments Q exhibits any of the levels of activity or potency at tor activation (or an IC50 for GIP receptor antagonism ) of 40 the GIP receptor described herein but has substantially less about 10 mM or less , or about 1 mM ( 1000 ?M ) or less ( e . g. , activity (potency ) at the glucagon receptor or the GLP - 1 about 750 uM or less , about 500 UM or less , about 250 uM receptor. In some embodiments, Q exhibits an EC so at the or less , about 100 uM or less , about 75 uM or less, about 50 glucagon receptor which is 100 - fold or greater than the EC50 uM or less , about 25 uM or less, about 10 uM or less, about at the GIP receptor and an EC5o at the GLP - 1 receptor which 7 . 5 uM or less , about 6 uM or less, about 5 uM or less , about 45 is 100 - fold or greater than the EC 5 , at the GIP receptor. 4 uM or less , about 3 uM or less , about 2 uM or less or about Activity at the GLP - 1 Receptor and the Glucagon Receptor 1 uM or less ). In some embodiments , the EC 50 or IC50 of Q In some embodiments , Q exhibits activity at both the at the GIP receptor is less than 1000 nM , less than 900 nM , GLP - 1 receptor and glucagon receptor (“ glucagon /GLP - 1 less than 800 nM , less than 700 nM , less than 600 nM , less receptor co - agonists ” ) . In some embodiments , the activity than 500 nM , less than 400 nM , less than 300 nM , or less 50 (e . g ., the EC50 or the relative activity or potency ) of Q at the than 200 nM . In some embodiments, the EC 50 or IC50 of Q glucagon receptor is within about 50 - fold , about 40 - fold , at the GIP receptor is about 100 nM or less , e .g ., about 75 about 30 - fold , about 20 - fold , about 10 -fold , or about 5 fold nM or less, about 50 nM or less, about 25 nM or less , about different (higher or lower) from its activity (e .g . , the EC 50 or 10 nM or less, about 8 nM or less , about 6 nM or less, about the relative activity or potency ) at the GLP - 1 receptor. In 5 nM or less , about 4 nM or less , about 3 nM or less , about 55 some embodiments , the glucagon potency of Q is within 2 nM or less , or about 1 nM or less . In some embodiments , about 25 - , about 20 -, about 15 -, about 10 - , or about 5 - fold the Q exhibits an EC50 or IC50 for GIP receptor activation different (higher or lower) from its GLP - 1 potency . which is in the picomolar range . In exemplary embodiments , In some embodiments , the ratio of the relative activity or the EC or ICs of Q at the GIP receptor is less than 1000 the EC 50 or the potency of the Q at the glucagon receptor PM , less than 900 PM , less than 800 PM , less than 700 PM , 60 divided by the relative activity or the EC 50 or potency of Q less than 600 PM , less than 500 PM , less than 400 PM , less at the GLP- 1 receptor is less than , or is about, X , wherein X than 300 PM , less than 200 pM . In some embodiments , the is selected from 100 , 75 , 60 , 50 , 40 , 30 , 20 , 15 , 10 , or 5 . In EC5o or IC50 of Q at the GIP receptor is about 100 PM or some embodiments, the ratio of the EC 50 or potency or less , e .g ., about 75 PM or less , about 50 PM or less , about relative activity of Q at the glucagon receptor divided by the 25 PM or less , about 10 PM or less , about 8 pM or less, about 65 EC50 or potency or relative activity of Q at the GLP - 1 6 PM or less, about 5 PM or less, about 4 PM or less, about receptor is about 1 less than 5 ( e . g ., about 4 , about 3 , about 3 PM or less , about 2 PM or less , or about 1 PM or less . 2 , about 1) . In some embodiments , the ratio of the glucagon US 9 , 783 ,592 B2 63 64 potency of Q compared to the GLP - 1 potency of Q is less receptor is within about 50 - fold , about 40 - fold , about than , or is about, Z , wherein Z is selected from 100 , 75 , 60 , 30 - fold , about 20 - fold , about 10 - fold , or about 5 fold dif 50 , 40 , 30 , 20 , 15 , 10 , and 5 . In some embodiments , the ratio ferent (higher or lower ) from its activity ( e . g ., the EC 50 or of the glucagon potency of compared to the GLP - 1 the relative activity or potency ) at the GLP - 1 receptor. In potency of Q is less than 5 ( e . g . , about 4 , about 3 , about 2 , 5 some embodiments , the GIP potency of Q is within about about 1 ) . In some embodiments , Q has an EC 50 at the 25 - , about 20 - , about 15 - , about 10 - , or about 5 - fold different glucagon receptor which is 2 - to 10 - fold ( e . g ., 2 - fold , 3 - fold , (higher or lower ) from its GLP - 1 potency . 4 - fold , 5 - fold , 6 - fold , 7 -fold , 8 - fold , 9 - fold , 10 - fold ) greater In some embodiments , the ratio of the relative activity or than the EC so at the GLP - 1 receptor. the EC 5 , or the potency of the Q at the GIP receptor divided In some embodiments , the ratio of the relative activity or 10 by the relative activity or the EC 50 or potency of Q at the potency or the EC of Q at the GLP - 1 receptor divided by GLP - 1 receptor is less than , or is about, X , wherein X is the relative activity or potency or the EC50 of the glucagon selected from 100 , 75 , 60 , 50 , 40 , 30 , 20 , 15 , 10 , or 5 . In analog at the glucagon receptor is less than , or is about, V , some embodiments , the ratio of the EC50 or potency or wherein V is selected from 100 , 75 , 60 , 50 , 40 , 30 , 20 , 15 , relative activity of Q at the GIP receptor divided by the EC 50 10 , or 5 . In some embodiments , the ratio of the EC 50 or 15 or potency or relative activity of Q at the GLP - 1 receptor is potency or relative activity of Q at the GLP - 1 receptor about 1 less than 5 ( e . g . , about 4 , about 3 , about 2 , about 1 ) . divided by the EC50 or potency or relative activity of Q at the In some embodiments , the ratio of the GIP potency of Q glucagon receptor is less than 5 ( e . g . , about 4 , about 3 , about compared to the GLP - 1 potency of Q is less than , or is about, 2 , about 1 ) . In some embodiments , the ratio of the GLP - 1 Z , wherein Z is selected from 100 , 75 , 60 , 50 , 40 , 30 , 20 , 15 , potency of compared to the glucagon potency of Q is less 2010 , and 5 . In some embodiments , the ratio of the GIP than , or is about, W , wherein W is selected from 100 , 75 , 60 , potency of Q compared to the GLP - 1 potency of Q is less 50 , 40 , 30 , 20 , 15 , 10 , and 5 . In some embodiments , the ratio than 5 ( e . g ., about 4 , about 3 , about 2 , about 1 ). In some of the GLP - 1 potency of a compared to the glucagon embodiments , Q has an EC50 at the GIP receptor which is 2 potency of Q is less than 5 (e . g ., about 4 , about 3 , about 2 , to 10 - fold ( e . g . , 2 - fold , 3 - fold , 4 - fold , 5 -fold , 6 - fold , 7 -fold , about 1 ) . In some embodiments , Q has an EC 50 at the GLP - 1 25 8 -fold , 9 - fold , 10 -fold ) greater than the EC so at the GLP - 1 receptor which is about 2 - to about 10 - fold ( e . g . , 2 - fold , receptor. 3 - fold , 4 - fold , 5 - fold , 6 - fold , 7 - fold , 8 - fold , 9 - fold , 10 - fold ) In some embodiments , the ratio of the relative activity or greater than the EC50 at the glucagon receptor. potency or the EC 50 of Q at the GLP - 1 receptor divided by In some embodiments , Q exhibits at least 0 . 1 % ( e . g ., the relative activity or potency or the EC so of Q at the GIP about 0 . 5 % or more , about 1 % or more , about 5 % or more , 30 receptor is less than , or is about, V , wherein V is selected about 10 % or more , ormore ) of the activity of native GLP - 1 from 100 , 75 , 60 , 50 , 40 , 30 , 20 , 15 , 10 , or 5 . In some at the GLP - 1 receptor (GLP - 1 potency ) and exhibits at least embodiments , the ratio of the EC50 or potency or relative 0 . 1 % ( e . g . , about 0 . 5 % or more , about 1 % or more , about 5 % activity of Q at the GLP - 1 receptor divided by the EC or or more , about 10 % or more , or more ) of the activity of potency or relative activity of Q at the GIP receptor is less native glucagon at the glucagon receptor (glucagon 35 than 5 ( e . g ., about 4 , about 3 , about 2 , about 1 ) . In some potency ). embodiments , the ratio of the GLP - 1 potency of compared Selectivity of Q for the glucagon receptor versus the to the GIP potency of Q is less than , or is about, W , wherein GLP - 1 receptor can be described as the relative ratio of Wis selected from 100 , 75 , 60 , 50 , 40 , 30 , 20 , 15 , 10 , and glucagon /GLP - 1 activity ( Q ' s activity at the glucagon recep - 5 . In some embodiments , the ratio of the GLP - 1 potency of tor relative to native glucagon , divided by the analog ' s 40 Q compared to the GIP potency of Q is less than 5 ( e . g . , activity at the GLP - 1 receptor relative to native GLP - 1 ) . For about 4 , about 3 , about 2 , about 1 ) . In some embodiments, example , a Q that exhibits 60 % of the activity of native Q has an EC 5 , at the GLP - 1 receptor which is about 2 - to glucagon at the glucagon receptor and 60 % of the activity of about 10 - fold ( e . g . , 2 - fold , 3 - fold , 4 - fold , 5 - fold , 6 - fold , native GLP - 1 at the GLP - 1 receptor has a 1 : 1 ratio of 7 - fold , 8 - fold , 9 - fold , 10 - fold ) greater than the EC50 at the glucagon /GLP - 1 activity . Exemplary ratios of glucagon 45 GIP receptor. GLP - 1 activity include about 1 : 1 , 1 . 5 : 1 , 2 : 1 , 3 : 1 , 4 : 1 , 5 : 1 , In some embodiments , Q exhibits at least 0 . 1 % ( e . g . , 6 : 1 , 7 : 1 , 8 : 1 , 9 : 1 or 10 : 1 , or about 1: 10 , 1 : 9 , 1 : 8 , 1 : 7 , 1 : 6 , about 0 . 5 % or more, about 1 % or more , about 5 % or more , 1 :5 , 1 : 4 , 1: 3 , 1 : 2, or 1 :1 .5 . As an example , a glucagon /GLP - 1 about 10 % or more , or more ) of the activity of native GLP - 1 activity ratio of 10 : 1 indicates a 10 - fold selectivity for the at the GLP - 1 receptor (GLP - 1 potency ) and exhibits at least glucagon receptor versus the GLP - 1 receptor. Similarly , a 50 0 . 1 % ( e . g . , about 0 . 5 % or more , about 1 % or more , about 5 % GLP - 1 /glucagon activity ratio of 10 : 1 indicates a 10 - fold or more , about 10 % or more , or more ) of the activity of selectivity for the GLP -1 receptor versus the glucagon native GIP at the GIP receptor (GIP potency ). receptor. Selectivity of Q for the GIP receptor versus the GLP - 1 In some embodiments , Q exhibits substantial activity receptor can be described as the relative ratio of GIP /GLP - 1 (potency ) at the glucagon receptor and GLP - 1 receptor and 55 activity ( Q ' s activity at the GIP receptor relative to native little to no activity at the GIP receptor . In some embodiments GIP , divided by the analog ' s activity at the GLP - 1 receptor Q exhibits any of the levels of activity or potency at the relative to native GLP - 1 ) . For example , a Q that exhibits glucagon receptor and the GLP - 1 receptor described herein 60 % of the activity of native GIP at the GIP receptor and but has substantially less activity ( potency ) at the GIP 60 % of the activity of native GLP - 1 at the GLP - 1 receptor receptor. In some embodiments , Q exhibits an EC50 at the 60 has a 1 : 1 ratio of GIP /GLP - 1 activity . Exemplary ratios of GIP receptor which is 100 - fold or greater than the EC50 at GIP /GLP - 1 activity include about 1 : 1 , 1 . 5 : 1 , 2 : 1 , 3 : 1 , 4 : 1 , the glucagon receptor and the EC 5 , at the GLP - 1 receptor. 5 : 1 , 6 : 1 , 7 : 1 , 8 : 1 , 9 : 1 or 10 : 1 , or about 1 : 10 , 1 : 9 , 1 : 8 , 1 : 7 , Activity at the GLP - 1 Receptor and the GIP Receptor 1 : 6 , 1 : 5 , 1 : 4 , 1 : 3 , 1 : 2 , or 1 : 1 . 5 . As an example, a GIP /GLP - 1 In some embodiments, exhibits activity at both the activity ratio of 10 : 1 indicates a 10 - fold selectivity for the GLP- 1 receptor and GIP receptor ( " GIP /GLP - 1 receptor 65 GIP receptor versus the GLP- 1 receptor. Similarly , a GLP co -agonists ” ) . In some embodiments , the activity ( e . g . , the 1 /GIP activity ratio of 10 : 1 indicates a 10 - fold selectivity for EC50 or the relative activity or potency ) of Q at the GIP the GLP - 1 receptor versus the GIP receptor. US 9 ,783 , 592 B2 65 66 Activity at the Glucagon Receptor and the GIP Receptor 1 : 8 , 1 :7 , 1 : 6 , 1 : 5 , 1 : 4 , 1 : 3 , 1 : 2 , or 1 : 1 . 5 . As an example , a In some embodiments , Q exhibits activity at both the GIP / glucagon activity ratio of 10 : 1 indicates a 10 - fold glucagon receptor and GIP receptor (“ GIP / glucagon recep - selectivity for the GIP receptor versus the glucagon receptor. tor co - agonists ” ) . In some embodiments , the activity ( e . g . , Similarly , a glucagon /GIP activity ratio of 10 : 1 indicates a the EC 5 , or the relative activity or potency ) of Q at the GIP 5 10 - fold selectivity for the glucagon receptor versus the GIP receptor is within about 50 - fold , about 40 - fold , about receptor. 30 - fold , about 20 - fold , about 10 - fold , or about 5 fold dif - Activity at the Glucagon Receptor, the GLP - 1 Receptor, and ferent (higher or lower ) from its activity (e . g. , the EC50 or the GIP Receptor the relative activity or potency ) at the glucagon receptor. In In some embodiments , Q exhibits activity at all three of some embodiments , the GIP potency of Q is within about 10 the glucagon receptor, the GLP - 1 receptor, and the GIP 25 - , about 20 -, about 15 - , about 10 - , or about 5 - fold different receptor (" glucagon /GLP - 1 /GIP receptor tri -agonists ” ) . In (higher or lower ) from its glucagon potency . some embodiments , the activity ( e . g . , the EC50 or the rela In some embodiments , the ratio of the relative activity or t ive activity or potency ) of Q at the glucagon receptor is the EC se or the potency of the Q at the GIP receptor divided within about 100 - fold , about 75 - fold , about 60 - fold , 50 - fold , by the relative activity or the EC50 or potency of Q at the 15 about 40 - fold , about 30 - fold , about 20 - fold , about 10 - fold , glucagon receptor is less than , or is about, X , wherein X is or about 5 fold different ( higher or lower ) from its activity selected from 100 , 75 , 60 , 50 , 40 , 30 , 20 , 15 , 10 , or 5 . In ( e . g ., the EC 50 or the relative activity or potency ) at both the some embodiments , the ratio of the EC50 or potency or GLP - 1 receptor and the GIP receptor. In some embodiments , relative activity of Q at the GIP receptor divided by the EC 50 the activity ( e . g . , the EC go or the relative activity or potency ) or potency or relative activity of Q at the glucagon receptor 20 of Q at the GLP - 1 receptor is within about 100 - fold , about is about 1 less than 5 ( e. g ., about 4 , about 3 , about 2 , about 75 - fold , about 60 - fold , 50 - fold , about 40 - fold , about 1) . In some embodiments , the ratio of the GIP potency of Q 30 - fold , about 20 - fold , about 10 -fold , or about 5 fold dif compared to the glucagon potency of Q is less than , or is ferent (higher or lower) from its activity ( e. g. , the EC50 or about, Z , wherein Z is selected from 100 , 75 , 60 , 50 , 40 , 30 , the relative activity or potency ) at both the glucagon recep 20 , 15 , 10 , and 5 . In some embodiments , the ratio of the GIP 25 tor and the GIP receptor. In some embodiments , the activity potency of Q compared to the glucagon potency of Q is less ( e . g ., the EC 50 or the relative activity or potency ) of Q at the than 5 ( e . g . , about 4 , about 3 , about 2 , about 1 ) . In some GIP receptor is within about 100 - fold , about 75 - fold , about embodiments , Q has an EC so at the GIP receptor which is 2 - 60 - fold , 50 - fold , about 40 - fold , about 30 - fold , about to 10 - fold ( e . g . , 2 - fold , 3 - fold , 4 - fold , 5 - fold , 6 - fold , 7 - fold , 20 - fold , about 10 - fold , or about 5 fold different (higher or 8 - fold , 9 - fold , 10 - fold ) greater than the EC 50 at the glucagon 30 lower ) from its activity ( e . g . , the EC 50 or the relative activity receptor . or potency ) at both the glucagon receptor and the GLP - 1 In some embodiments , the ratio of the relative activity or receptor. This fold difference can be alternatively expressed potency or the EC 50 of Q at the glucagon receptor divided by as ratios of glucagon /GLP - 1 , or GLP - 1 /GIP , or glucagon / the relative activity or potency or the EC50 of Q at the GIP GLP - 1 as above . receptor is less than , or is about, V , wherein V is selected 35 Structure of the Glucagon Superfamily Peptide ( Q ) from 100 , 75 , 60 , 50 , 40 , 30 , 20 , 15 , 10 , or 5 . In some The glucagon superfamily peptide ( Q ) described herein embodiments , the ratio of the EC50 or potency or relative can comprise an amino acid sequence which is based on the activity of Q at the glucagon receptor divided by the EC50 amino acid sequence of native human glucagon (SEQ ID or potency or relative activity of Q at the GIP receptor is less NO : 1601) , native human GLP - 1 ( SEQ ID NOs: 1603 or than 5 ( e . g . , about 4 , about 3 , about 2 , about 1 ) . In some 40 1604 ) , or native human GIP (SEQ ID NO : 1607) . embodiments , the ratio of the glucagon potency of Q com - Based on Native Human Glucagon pared to the GIP potency of Q is less than , or is about , W , In some aspects of the invention , the glucagon superfam wherein W is selected from 100 , 75 , 60 , 50 , 40 , 30 , 20 , 15 , ily peptide ( Q ) comprises an amino acid sequence that is 10 , and 5 . In some embodiments , the ratio of the glucagon based on the amino acid sequence of native human glucagon potency of Q compared to the GIP potency of Q is less than 45 (SER ID NO : 1601) . In some aspects , Q comprises a 5 ( e . g . , about 4 , about 3 , about 2 , about 1 ) . In some modified amino acid sequence of SEQ ID NO : 1601 com embodiments , Q has an EC50 at the glucagon receptor which prising 1 , 2 , 3 , 4 , 5 , 6 , 7 , 8 , 9 , 10 , 11 , 12 , 13 , 14 , 15 , and in is about 2 - to about 10 - fold ( e . g . , 2 - fold , 3 - fold , 4 - fold , some instances, 16 or more (e . g ., 17 , 18 , 19, 20 , 21 , 22 , 23 , 5 - fold , 6 - fold , 7 - fold , 8 - fold , 9 - fold , 10 - fold ) greater than 24 , 25 , etc . ) , amino acid modifications. In some embodi the EC - o at the GIP receptor . 50 ments , Q comprises a total of 1 , up to 2 , up to 3 , up to 4 , up In some embodiments , Q exhibits at least 0 . 1 % ( e . g ., to 5 , up to 6 , up to 7 , up to 8 , up to 9 , or up to 10 amino acid about 0 . 5 % or more , about 1 % or more , about 5 % or more , modifications relative to the native human glucagon about 10 % or more , or more ) of the activity of native sequence (SEQ ID NO : 1601 ) . In some embodiments , the glucagon at the glucagon receptor (glucagon potency ) and modifications are any of those described herein , e . g . , acy exhibits at least 0 . 1 % ( e . g . , about 0 . 5 % or more , about 1 % 55 lation , alkylation , pegylation , truncation at C - terminus , sub or more , about 5 % or more , about 10 % or more , or more ) of stitution of the amino acid at one or more of positions 1 , 2 , the activity of native GIP at the GIP receptor (GIP potency ) . 3 , 7 , 10 , 12 , 15 , 16 , 17 , 18 , 19 , 20 , 21 , 23 , 24 , 27 , 28 , and Selectivity of Q for the GIP receptor versus the glucagon 29 . receptor can be described as the relative ratio of GIP / In some embodiments , comprises an amino acid glucagon activity ( Q ' s activity at the GIP receptor relative to 60 sequence which has at least 25 % sequence identity to the native GIP , divided by the analog ' s activity at the glucagon amino acid sequence of native human glucagon (SEQ ID receptor relative to native glucagon ) . For example , a Q that NO : 1601) . In some embodiments, Q comprises an amino exhibits 60 % of theactivity of native GIP at the GIP receptor acid sequence which is at least 30 % , at least 40 % , at least and 60 % of the activity of native glucagon at the glucagon 50 % , at least 60 % , at least 70 % , at least 80 % , at least 85 % , receptor has a 1 : 1 ratio of GIP / glucagon activity . Exemplary 65 at least 90 % or has greater than 90 % sequence identity to ratios of GIP / glucagon activity include about 1 : 1 , 1 . 5 : 1 , 2 : 1 , SEQ ID NO : 1601. In some embodiments, the amino acid 3 : 1 , 4 : 1 , 5 : 1 , 6 : 1 , 7 : 1 , 8 : 1 , 9 : 1 or 10 : 1 , or about 1 : 10 , 1 : 9 , sequence of Q , which has the above - referenced % sequence US 9 , 783 ,592 B2 68 identity is the full -length amino acid sequence of Q . In some limited to , e. g. , wherein A is 90 % and C and D are 1 and 27 , embodiments , the amino acid sequence of which has the or 6 and 27 , or 8 and 27 , or 10 and 27 , or 12 and 27 , or 16 above - referenced % sequence identity is only a portion of and 27 . the amino acid sequence of 0 . In some embodiments , O Based on Native Human GIP comprises an amino acid sequence which has about A % or 5 In some embodiments of the present disclosures, Q is an greater sequence identity to a reference amino acid sequence analog of native human GIP , the amino acid sequence of which is provided herein as SEQ ID NO : 1607 . Accordingly , of at least 5 contiguous amino acids ( e . g ., at least 6 , at least in some embodiments , Q comprises an amino acid sequence 7 , at least 8 , at least 9 , at least 10 amino acids) of SEQ ID which is based on the amino acid sequence of SEQ ID NO : NO : 1601 , wherein the reference amino acid sequence 10 1607 but is modified with 1 , 2 , 3 , 4 , 5 , 6 , 7 , 8 , 9 , 10 , 11 , 12 , begins with the amino acid at position C of SEQ ID NO : 13 , 14 , 15 , and in some instances , 16 or more ( e . g . , 17 , 18 , 1601 and ends with the amino acid at position D of SEQ ID 19, 20 , 21, 22 , 23, 24 , 25, etc .) , amino acid modifications. In NO : 1601 , wherein A is 25 , 30 , 35 , 40 , 45 , 50 , 55 , 60 , 65 , some embodiments , Q comprises a total of 1 , up to 2 , up to 70 , 75 , 80 , 85 , 90 , 91, 92 , 93 , 94 , 95 , 96 , 97 , 98 , 99 ; C is 1 , 3 , up to 4 , up to 5 , up to 6 , up to 7 , up to 8 , up to 9 , or up 2 , 3 , 4 , 5 , 6 , 7 , 8 , 9 , 10 , 11, 12 , 13 , 14 , 15 , 16 , 17 , 18 , 1919 , 15 to 10 amino acid modifications relative to the native human 20 , 21, 22 , 23 , 24 , 25 , 26 , 27 , or 28 and D is 5 , 6 , 7 , 8 , 9 , GIP sequence (SEQ ID NO : 1607 ) . In some embodiments , 10 , 11 , 12 , 13 , 14 , 15 , 16 , 17 , 18 , 19 , 20 , 21 , 22 , 23 , 24 , 25 , the modifications are any of those described herein . eg . 26 , 27, 28 or 29 . Any and all possible combinations of the acylation , alkylation , pegylation , truncation at C - terminus , foregoing parameters are envisioned , including but not substitution of the amino acid at one or more of positions 1 , limited to , e . g ., wherein A is 90 % and C and D are 1 and 27, 20 2 , 3 , 7 , 10, 12 , 15, 16 , 17, 18, 19 , 20 , 21 , 23, 24 , 27 , 28 , and or 6 and 27 , or 8 and 27 , or 10 and 27 , or 12 and 27 , or 16 29 . Exemplary GIP receptor agonists are known in the art . and 27 . See , for example , Irwin et al . , J Pharm and Expmt Ther Based on Native Human GLP - 1 314 ( 3 ) : 1187 - 1194 ( 2005 ) ; Salhanick et al. , Bioorg Med In some aspects of the invention , the glucagon superfam - Chem Lett 15 ( 18 ) : 4114 -4117 ( 2005 ) ; Green et al. , Dibetes ily peptide ( Q ) comprises an amino acid sequence that is 25 7 (5 ) : 595 -604 ( 2005 ) ; O 'Harte et al. , J Endocrinol 165 ( 3 ) : based on the amino acid sequence of native human GLP - 1 639 -648 ( 2000 ) ; O 'Harte et al. , Diabetologia 45 ( 9 ) : 1281 ( SEO ID NO : 1603 ) . In some aspects , 0 comprises a 1291 ( 2002 ) ; Gault et al. , Biochem J 367 ( Pt3 ) : 913 - 920 ( 2002) ; Gault et al ., J Endocrin 176 : 133 - 141 (2003 ) ; Irwin modified amino acid sequence of SEQ ID NO : 1603 com et al. , Diabetes Obes Metab. 11 (6 ): 603 -610 (epub 2009 ) . prising 1 , 2 , 3 , 4 , 5 , 6 , 7 , 8 , 9 , 10 , 11, 12, 13 , 14 , 15, and23 in 30 In some embodiments , comprises an amino acid some instances, 16 or more (e .g ., 17 , 18, 19, 20 , 21, 22 , 23 , sequence which has at least 25 % sequence identity to the 24 , 25 , etc .) , amino acid modifications . In some embodi amino acid sequence of native human GIP (SEQ ID NO : ments, Q comprises a total of 1 , up to 2 , up to 3 , up to 4 , up 1607 ) . In some embodiments , Q comprises an amino acid to 5 , up to 6 , up to 7 , up to 8 , up to 9 , or up to 10 amino acid sequence which is at least 30 % , at least 40 % , at least 50 % , modifications relative to the native human GLP - 1 sequence 35 at least 60 % , at least 70 % , at least 80 % , at least 85 % , at least (SEQ ID NO : 1603 ) . In some embodiments , the modifica 90 % or has greater than 90 % sequence identity to SEO ID tions are any of those described herein , e . g ., acylation , NO : 1607 . In some embodiments , the amino acid sequence alkylation , pegylation , truncation at C -terminus , substitution of which has the above- referenced % sequence identity is of the amino acid at one or more of positions 1 , 2 , 3 , 7 , 10 , the full - length amino acid sequence of Q . In some embodi 12 , 15 , 16 , 17 , 18 , 19 , 20 , 21 , 23 , 24 , 27 , 28 , and 29 . 40 ments , the amino acid sequence of Q which has the above In some embodiments , comprises an amino acid referenced % sequence identity is only a portion of the sequence which has at least 25 % sequence identity to the amino acid sequence of Q . In some embodiments , Q com amino acid sequence of native human GLP - 1 (SEQ ID NO : prises an amino acid sequence which has about A % or 1603 ) . In some embodiments , Q comprises an amino acid greater sequence identity to a reference amino acid sequence sequence which is at least 30 % , at least 40 % , at least 50 % , 45 of at least 5 contiguous amino acids ( e . g ., at least 6 , at least at least 60 % , at least 70 % , at least 80 % , at least 85 % , at least 7 , at least 8 , at least 9 , at least 10 amino acids ) of SEQ ID 90 % or has greater than 90 % sequence identity to SEQ ID NO : 1607 , wherein the reference amino acid sequence NO : 1603. In some embodiments , the amino acid sequence begins with the amino acid at position C of SEQ ID NO : of Q , which has the above -referenced % sequence identity is 1607 and ends with the amino acid at position D of SEQ ID the full- length amino acid sequence of Q . In some embodi - 50 NO : 1607, wherein A is 25 , 30 , 35 , 40 , 45 , 50, 55 , 60 , 65 , ments , the amino acid sequence of Q which has the above 70 , 75 , 80 , 85 , 90 , 91, 92 , 93 , 94 , 95 , 96 , 97 , 98 , 99 ; C is 1 , referenced % sequence identity is only a portion of the 2 , 3 , 4 , 5 , 6 , 7 , 8 , 9 , 10 , 11 , 12 , 13 , 14 , 15 , 16 , 17 , 18 , 19 , amino acid sequence of Q . In some embodiments, Q com - 20 , 21, 22 , 23 , 24 , 25 , 26 , 27 , or 28 and D is 5 , 6 , 7 , 8 , 9 , prises an amino acid sequence which has about A % or 10 , 11 , 12 , 13 , 14 , 15 , 16 , 17 , 18 , 19, 20 , 21, 22 , 23 , 24 , 25 , greater sequence identity to a reference amino acid sequence 55 26 , 27 , 28 or 29 . Any and all possible combinations of the of at least 5 contiguous amino acids ( e . g . , at least 6 , at least foregoing parameters are envisioned , including but not 7 , at least 8 , at least 9 , at least 10 amino acids ) of SEQ ID limited to , e . g . , wherein A is 90 % and C and D are 1 and 27 , NO : 1603 , wherein the reference amino acid sequence or 6 and 27 , or 8 and 27 , or 10 and 27 , or 12 and 27 , or 16 begins with the amino acid at position C of SEQ ID NO : and 27 . 1603 and ends with the amino acid at position D of SEQ ID 60 Modifications NO : 1603 , wherein A is 25 , 30 , 35 , 40 , 45 , 50 , 55 , 60 , 65 , Where is a glucagon related peptide , O can comprise 70 , 75 , 80 , 85 , 90 , 91 , 92 , 93 , 94 , 95 , 96 , 97 , 98 , 99 ; C is 1 , the native glucagon amino acid sequence ( SEQ ID NO : 2 , 3 , 4 , 5 , 6 , 7 , 8 , 9 , 10 , 11 , 12, 13 , 14 , 15, 16 , 17 , 18 , 19, 1601 ) with modifications. In exemplary embodiments , the 20 , 21, 22 , 23 , 24 , 25 , 26 , 27 , or 28 and D is 5 , 6 , 7 , 8 , 9 , glucagon related peptide may comprise a total of 1 , up to 2 , 10 , 11 , 12 , 13 , 14 , 15 , 16 , 17 , 18 , 19 , 20 , 21 , 22 , 23 , 24 , 25 , 65 up to 3 , up to 4 , up to 5 , up to 6 , up to 7 , up to 8 , up to 9 , 26 , 27 , 28 or 29 . Any and all possible combinations of the or up to 10 amino acid modifications relative to the native foregoing parameters are envisioned , including but not glucagon sequence , e . g . conservative or non -conservative US 9 , 783 ,592 B2 69 substitutions. Modifications and substitutions described herein are , in certain aspects made at specific positions Structure I within wherein the numbering of the position corresponds to the numbering of glucagon (SEQ ID NO : 1601) . In some wat R1 - CH2 - X - 1 R2 embodiments 1, 2 , 3 , 4 or 5 non - conservative substitutions 5 axi , are carried out at any of positions 2 , 5 , 7 , 10 , 11 , 12 , 13con , 14 , Structure II 17 , 18 , 19 , 20 , 21, 24 , 27 , 28 or 29 and up to 5 further conservative substitutions are carried out at any of these positions. In some embodiments 1, 2 , or 3 amino acid modifications are carried outwithin amino acids at positions fuel, 1 - 16 , and 1 , 2 or 3 amino acid modifications are carried out Structure III within amino acids at positions 17 - 26 . In some embodi ments , Q retains at least 22 , 23 , 24 , 25 , 26 , 27 or 28 of the 5 – R – CH2 – – CH2 = R * naturally occurring amino acids at the corresponding posi- 15 muutos - Lan tions in native glucagon ( e . g . have 1 - 7 , 1 - 5 or 1 - 3 modifi cations relative to naturally occurring glucagon ) . wherein R is Co- 3 alkyl or Co- 3 heteroalkyl; R² is NHR + or DPP - IV Resistance C1- 3 alkyl; Rºis C / - 3 alkyl; R * is H or C1- 3 alkyl; X is NH , In some embodiments , where Q is a glucagon superfamily O , or S ; and Y is NHR4, SR " , or OR3. In some embodiments , peptide , O comprises a modification at position 1 or 2 to 20 X is NH or Y is NHR4 In some embodiments . Rt is C . reduce susceptibility to cleavage by dipeptidyl peptidase IV alkyl or C , heteroalkyl . In some embodiments , R2 is NHR + ( DPP - IV ) . More particularly , in some embodiments , posi or C , alkyl. In some embodiments , R4 is H or C1 alkyl. In tion 1 of Q (e . g. , selected from those in FIG . 1) is substituted exemplary embodiments in which Q is a Class 1 , Class 2 , or with an amino acid selected from the group consisting of Class 3 glucagon related peptide, an amino acid comprising D -histidine , alpha , alpha -dimethyl imidiazole acetic acid 25 a side chain of Structure I is provided where , R ' is CH , S , (DMIA ), N -methyl histidine, alpha -methyl histidine , imida X is NH , and R2 is CHz ( acetamidomethyl- cysteine , zole acetic acid , desaminohistidine , hydroxyl- histidine, C (Acm ) ); R ' is CH?, X is NH , and R² is CHz (acetyldiamin acetyl- histidine and homo -histidine . More particularly , in obutanoic acid , Dab (Ac ) ); R is C , alkyl, X is NH , R2 is some embodiments , position 2 of Q is substituted with an NHR4, and R4 is H (carbamoyldiaminopropanoic acid , Dap amino acid selected from the group consisting of D -serine , (urea ) ); or RP is CH , CH2, X is NH , and R2 is CHz D - alanine, valine, glycine, N -methyl serine, and amin (acetylornithine , Orn ( Ac ) ) . In exemplary embodiments an oisobutyric acid . In some embodiments , position 2 of the amino acid comprising a side chain of Structure II is glucagon related peptide is not D - serine . provided where , Rl is CH2, Y is NHR4, and R4 is CHz Glucagon Modification at Position 3 35 (methylglutamine , Q (Me )) ; In exemplary embodiments an Glucagon related peptides of Classes 1 to 3 described amino acid comprising a side chain of Structure IIII is herein may bemodified atposition 3 (according to the amino provided where , R1 is CH , and R4 is H (methionine - sulfox acid numbering of wild type glucagon ) to maintain or ide , M ( O ) ) ; In specific embodiments , the amino acid at increase activity at the glucagon receptor. position 3 is substituted with Dab ( Ac ) . In some embodiments in which Q is a Class 1 , Class 2 , or 40 Acylation of O Class 3 glucagon related peptide , maintained or enhanced In some embodiments, the glucagon related peptide ( e . g . activity at the glucagon receptor may be achieved by modi - a Class 1 glucagon related peptide , Class 2 glucagon related fying the Gln at position 3 with a glutamine analog . For peptide, Class 3 glucagon related peptide , Class 4 glucagon example, a Class 1 , Class 2 , or Class 3 glucagon related related peptide , Class 4 glucagon related peptides or Class 5 peptide comprising a glutamine analog at position 3 may 45 glucagon related peptide) , Q is modified to comprise an acyl exhibit about 5 % , about 10 % , about 20 % , about 50 % , or group . The acyl group can be covalently linked directly to an about 85 % or greater the activity of native glucagon (SEQ amino acid of the peptide Q , or indirectly to an amino acid ID NO : 1601) at the glucagon receptor . In some embodi - of via a spacer , wherein the spacer is positioned between ments a Class 1 , Class 2 , or Class 3 glucagon related peptide the amino acid of Q and the acyl group . Q may be acylated comprising a glutamine analog at position 3 may exhibit 50 at the same amino acid position where a hydrophilic moiety about 20 % , about 50 % , about 75 % , about 100 % , about is linked , or at a different amino acid position . As described 200 % or about 500 % or greater the activity of a correspond herein , Q can be a glucagon superfamily peptide , glucagon ing glucagon peptide having the same amino acid sequence related peptide, including a Class 1 , 2 , 3 , 4 or 5 glucagon as the peptide comprising the glutamine analog, except for related peptide , or osteocalcin , calcitonin , amylin , or an the modified amino acid at position 3 at the glucagon 55 analog , derivative or conjugate thereof. For example , Q may receptor. In some embodiments , a Class 1 , Class 2 , or Class be one of Class 1 , Class 2 , Class 3 , Class 4 or Class 5 , and 3 glucagon related peptide comprising a glutamine analog at may comprise an acyl group which is non -native to a position 3 exhibits enhanced activity at the glucagon recep - naturally - occurring amino acid . Acylation can be carried out tor, but the enhanced activity is no more than 1000 % , at any position within Q . Where is a glucagon related 10 , 000 % , 100 , 000 % , or 1 , 000 ,000 % of the activity of native 60 peptide , acylation may occur at any position including any glucagon or of a corresponding glucagon related peptide of positions 1 - 29 , a position within a C - terminal extension , having the same amino acid sequence as the peptide com - or the C - terminal amino acid , provided that the activity prising the glutamine analog , except for the modified amino exhibited by the non - acylated glucagon related peptide is acid at position 3 . retained upon acylation . For example , if the unacylated In some embodiments , the glutamine analog is a naturally 65 peptide has glucagon agonist activity , then the acylated occurring or a non - naturally occurring amino acid compris - peptide retains the glucagon agonist activity . Also for ing a side chain of Structure I, II or III : example , if the unacylated peptide has glucagon antagonist US 9 , 783 ,592 B2 71 activity , then the acylated peptide retains the glucagon antagonist activity . For instance , if the unacylated peptide [ Formula II ] has GLP - 1 agonist activity , then the acylated peptide retains H2N - C - COOH GLP - 1 agonist activity . Nonlimiting examples include acy lation at positions 5 , 7 , 10 , 11, 12, 13, 14 , 16 , 17 , 18 , 19 , 20 , 5 (CH2 ) n 21 , 24, 27 , 28 , or 29 ( according to the amino acid numbering of wild type glucagon ). With regard to Class 1 , Class 2 , and OH Class 3 glucagon related peptides, acylation may occur at wherein n = 1 to 4 any of positions 5 , 7 , 10 , 11 , 12 , 13 , 14 , 16 , 17 , 18 , 19 , 20 , 21, 24 , 27 , 28 , 29, 30 , 37 , 38 , 39, 40 , 41, 42 , or 43 ( according 10 In some exemplary embodiments , the amino acid of For to the amino acid numbering of wild type glucagon ). Other mula II is the amino acid wherein n is 1 ( Ser ) . nonlimiting examples with respect to glucagon related pep . In yet other embodiments , the amino acid of peptide Q tides ( e. g . , Class 1 , 2 , 3 , 4 , or 5 ) include acylation at position comprising a side chain thiol is an amino acid of Formula 10 (according to the amino acid numbering of the wild type 15 NI: glucagon ) and pegylation at one or more positions in the C - terminal portion of the glucagon peptide , e . g ., position 24 , 28 or 29 ( according to the amino acid numbering of the wild [Formula III ] type glucagon ), within a C -terminal extension , or at the HN – – COOH C -terminus ( e .g ., through adding a C - terminal Cys ) . 20 (CH2 ) n In a specific aspect of the invention , peptide Q ( e . g ., a glucagon superfamily peptide , a glucagon related peptide, a SH Class 1 , 2 , 3 , 4 or 5 glucagon related peptide, or osteocalcin , wherein n = 1 to 4 calcitonin , amylin , or an analog, derivative or conjugate thereof) is modified to comprise an acyl group by direct - In some exemplary embodiments , the amino acid of For acylation of an amine , hydroxyl, or thiol of a side chain of mula III is the amino acid wherein n is 1 (Cys ) . an amino acid of Q . In some embodiments , Q is directly In yet other embodiments , the amino acid of peptide Q acylated through the side chain amine, hydroxyl , or thiol of comprising a side chain amine, hydroxyl, or thiol is a an amino acid . In some embodiments , where Q is a glucagon 30 disubstituted amino acid comprising the same structure of related peptide , acylation is at position 10 , 20 , 24 , or 29 Formula I, Formula II, or Formula III, except that the ( according to the amino acid numbering of the wild type hydrogen bonded to the alpha carbon of the amino acid of glucagon ) . In this regard , the acylated glucagon related Formula 1 , Formula II , or Formula III is replaced with a peptide can comprise the amino acid sequence of SEQ ID second side chain . NO : 1601, or a modified amino acid sequence thereof 35 In some embodiments of the invention , the acylated comprising one or more of the amino acid modifications peptide OlegQ (e . g ., aa glucagonglucagon superfamily peptide , a glucagon described herein , with at least one of the amino acids at positions 10 , 20 , 24 , and 29 ( according to the amino acid related peptide , a Class 1 , 2 , 3 , 4 or 5 glucagon related numbering of the wild type glucagon ) modified to any amino peptide, or osteocalcin , calcitonin , amylin , or an analog , acid comprising a side chain amine, hydroxyl, or thiol . In 40 derivative or conjugate thereof) comprises a spacer between some specific embodiments of the invention , where Q is a 40 the peptide and the acyl group . In some embodiments , Q is glucagon related peptide , the direct acylation of the Q occurs covalently bound to the spacer , which is covalently bound to through the side chain amine , hydroxyl, or thiol of the amino the acyl group . In some exemplary embodiments , Q is acid at position 10 (according to the amino acid numbering modified to comprise an acyl group by acylation of an of the wild type glucagon ). 45is amine, hydroxyl, or thiol of a spacer , which spacer (where In some embodiments , the amino acid of peptide Q ( e. g. , Q is a glucagon related peptide, e. g ., Class 1 , 2 , 3, 4 or 5 ) a glucagon superfamily peptide, a glucagon related peptide, is attached to a side chain of an amino acid at position 10 , a Class 1 , 2 , 3 , 4 or 5 glucagon related peptide , or osteo 20 , 24 , or 29 ( according to the amino acid numbering of the calcin , calcitonin , amylin , or an analog , derivative or con wild type glucagon ), or at the C -terminal amino acid of the jugate thereof) comprising a side chain amine is an amino 50 glucagon related peptide . The amino acid of peptide o to acid of Formula I : which the spacer is attached can be any amino acid com prising a moiety which permits linkage to the spacer. For example , an amino acid comprising a side chain — NH2, [ Formula I ] OH , or — COOH ( e . g . , Lys, Orn , Ser, Asp , or Glu ) is 55 suitable . An amino acid ofpeptide Q ( e. g ., a singly or doubly H?N – – COOH a - substituted amino acid ) comprising a side chain - NH2, (CH2 )n OH , or — COOH ( e . g . , Lys , Orn , Ser, Asp , or Glu ) is also suitable. In some embodiments where is a glucagon NH2 related peptide ( e. g ., Class 1, 2, 3 , 4 or 5 ), the acylated wherein n = 1 to 4 60 glucagon related peptide can comprise the amino acid sequence of SEQ ID NO : 1601 , or a modified amino acid sequence thereof comprising one or more of the amino acid In some exemplary embodiments , the amino acid of For modifications described herein , with at least one of the mula I, is the amino acid wherein n is 4 (Lys ) or n is 3 (Orn ). amino acids at positions 10 , 20 , 24 , and 29 (according to the In other embodiments , the amino acid of peptide Q 65 amino acid numbering of the wild type glucagon ) modified comprising a side chain hydroxyl is an amino acid of to any amino acid comprising a side chain amine , hydroxyl, Formula II: or carboxylate .