USOO8685916B2

(12) United States Patent (10) Patent No.: US 8,685,916 B2 Jenkins et al. (45) Date of Patent: Apr. 1, 2014

(54) OPOD PRODRUGS WITH HETEROCYCLC WO 20081012O2 8, 2008 LINKERS WO 2010.045599 4/2010 WO 2011 133346 4/2011 (75) Inventors: Thomas E. Jenkins, San Carlos, CA (US); Craig O. Husfeld, San Carlos, CA OTHER PUBLICATIONS (US) Berkop-Schnurch “The use of inhibitory agents to overcome the enzymatic barrier to perorally administered therapeutic peptides and (73) Assignee: Signature Therapeutics, Inc., Palo Alto, proteins' J. Control. Release (1998), vol. 50, No. 1-2, pp. 1-16. CA (US) Birk et al., “Trypsin and chymotrypsin inhibitors from soybeans' Methods in Enzymology (1976) vol. 45, pp. 700-707. (*) Notice: Subject to any disclaimer, the term of this Geratz et al., “Novel bis(benzamidine) compounds with an aromatic patent is extended or adjusted under 35 central link. Inhibitors of thrombin, pancreatic kallikrein, trypsin, U.S.C. 154(b) by 0 days. and complement” J. Med. Chem. (1976), vol. 19, pp. 634-639. Göke et al., “Effect of a Specific Serine Protease Inhibitor on the Rat (21) Appl. No.: 13/415,790 Pancreas: Systemic Administration of Camostate and Exocrine” Digestion (1984) vol. 30, pp. 171-178. (22) Filed: Mar. 8, 2012 Gomes et al., “Cyclization-activated prodrugs' Molecules, (2007), vol. 12, pp. 2484-2506. (65) Prior Publication Data Hijikata et al., “Selective Inhibition of Trypsin by (2R-4R)-4- Phenyl-1-NO-(7-methoxy-2-naphthalenesulfonyl)-L-arginyl-2- US 2012/O295834 A1 Nov. 22, 2012 piperidinecarboxylic Acid' J. Biochem. (2000), vol. 275, pp. 18995 18999. Related U.S. Application Data Kunze et al., “Effects of the serine protease inhibitors FOY and FOY 305 on phospholipase A I (EC 3.1.1.32) activity in rat—liver (60) Provisional application No. 61/451,010, filed on Mar. lysosomes' Pharm. Research Com. (1983), vol. 15, pp. 451–459. 9, 2011, provisional application No. 61/583,516, filed Lin et al., “The 0.25-nm X-ray structure of the Bowman-Birk type on Jan. 5, 2012. inhibitor from mung bean internary complex with porcine trypsin' Eur, J. Biochem. (1993), vol. 212, pp. 549-555. (51) Int. C. Markwardt et al., "Comparative studies on the inhibition of trypsin, A638/09 (2006.01) plasmin, and thrombin by derivatives of benzylamine and A6 IK 47/00 (2006.01) benzylamidine” Eur, J. Biochem, (1968), vol. 6, pp. 502-506. (52) U.S. C. Ozawa et al., “The reactive site of trypsin inhibitors' J. Biol. Chem. USPC ...... 514/1.1 : 514/1.3 (1966), vol. 241, pp. 3955-3961. Ramjee et al. “The Kinetic and Structural Characterization of the (58) Field of Classification Search Reaction of Nafamostat with Bovine Pancreatic Trypsin' Thrmb Res. None (2000), vol. 98, No. 6, pp. 559-569. See application file for complete search history. Renatus et al. “Structural and Functional Analyses of Benzamidine Based Inhibitors in Complex with Trypsin: Implications for the Inhi (56) References Cited bition of Factor Xa, tRA, and Urokinase” J. Med. Chem... (1998), vol. 41, No. 27 pp. 5445-5456. U.S. PATENT DOCUMENTS Tanizawa et al. “Inverse Substrates for Tryspin and Tryspin-like Enzymes' Acc. Chem. Res., (1987), vol. 20, pp. 337-343. 4454,338 A 6, 1984 Fujii et al. 4,532,255 A 7, 1985 Fujii et al. Testa et al. “Hydrolysis in Drug and Prodrug Metabolism” Verlag 5,217.987 A * 6, 1993 Berger ...... 514,416 Helvetica Chimica Acta, Postfach, CH-8042, Switzerland (2003) pp. 5,352,704 A 10, 1994 Okuyama et al. 420-534. 6,245,802 B1* 6, 2001 Iyengaret al...... 514,438 6,388,122 B1 5, 2002 Kido et al. (Continued) 6,586, 196 B1 T/2003 Bronstein et al. 7,060,290 B1 6, 2006 Morimoto et al. 7,189,414 B2 3, 2007 Rubinstein et al. Primary Examiner — Marcella M Cordero Garcia 7,893,105 B2 2, 2011 Xiang et al. Assistant Examiner — Sergio Coffa 2003/018O352 A1 9, 2003 Patel et al. 2004, OO63628 A1 4, 2004 Piccariello et al. (74) Attorney, Agent, or Firm — Carol L. Francis; Khin K. 2005/0176644 A1 8, 2005 Mickle et al. Chin; Bozicevic, Field & Francis LLP 2007/0O82929 A1 4, 2007 Gant et al. 2007, 0123468 A1 5/2007 Jenkins et al. 2007/02O3055 A1 8, 2007 Mickle et al. (57) ABSTRACT 2009,0136980 A1 5/2009 Bebbington et al. 2009.0137618 A1 5/2009 Jenkins et al. The embodiments provide prodrug compounds of Formulae 2009,0192093 A1 T/2009 Mickle et al. I-XV. The present disclosure also provides compositions, and 2009/0209569 A1 8, 2009 Arnelle et al. their methods of use, where the compositions comprise a 2010.0035826 A1 2, 2010 Jenkins et al. 2010, 0080797 A1 4, 2010 Yeomans et al. prodrug compound of Formulae I-XV that provides con 2010/0092562 A1 4, 2010 Hollenbeck et al. trolled release of an opioid. Such compositions can optionally 2010/0286.186 A1 11, 2010 Franklin et al. provide a trypsin inhibitor that interacts with the enzyme that mediates the controlled release of an opioid from the prodrug FOREIGN PATENT DOCUMENTS So as to attenuate enzymatic cleavage of the prodrug.

WO 2007 140272 12/2007 WO 2008.101187 8, 2008 123 Claims, 26 Drawing Sheets US 8,685,916 B2 Page 2

(56) References Cited hospital inpatients' Clinical Pharmacology and Therapeutics, (2004) vol. 76, No. 6, pp. 639-647. Umezawa et al., “Structure and activities of protease inhibitors of OTHER PUBLICATIONS microbial origin' Methods in Enzymology (1976) vol. 45, pp. 678 695. Tirkkonen et al., “Drug interactions with the potential to prevent prodrug activation as a common source of irrational prescribing in * cited by examiner U.S. Patent Apr. 1, 2014 Sheet 1 of 26 US 8,685,916 B2

Figure 1

rax

Eihilist U.S. Patent Apr. 1, 2014 Sheet 2 of 26 US 8,685,916 B2

Figure 2

38ts

SSS

Srs U.S. Patent Apr. 1, 2014 Sheet 3 of 26 US 8,685,916 B2

Figure 2 (continued)

3S 8.

U.S. Patent Apr. 1, 2014 Sheet 4 of 26 US 8,685,916 B2

Figure 3

&SF& A .. &ra:S

H H ESS SSSSS &

F3s. P&FE: :

s $ . & s & ......

Hs s E. t & Hs & s S: S. NSSSSSS $83. Lisse ESs 3S

s is m site s

. ::$ter SSS Essie & lists SSSSS is: U.S. Patent Apr. 1, 2014 Sheet 5 of 26 US 8,685,916 B2

Figure 4 OC from Oxycodone (28 umol/kg) OC from Compound KC-9 (31 umol/kg) OC from Compound KC-11 (34 umol/kg ) 1OO OC from Compound KC-12 (29 umol/kg ) OC from Compound KC-13 (29 umol/kg ) OC from Compound KC-14 (29 umol/kg) OC from Compound KC-15 (29 umol/kg ) OC from Compound KC-16 (29 umol/kg OC from Compound KC-17 (28 umol/kg

Time, hr U.S. Patent Apr. 1, 2014 Sheet 6 of 26 US 8,685,916 B2

Figure 5A OC from Oxycodone (5.7 umol/kg) OC from 20 mg OxyContin Tablet OC from Compound KC-12 (5.7 umol/kg) OC from Compound KC-13 (5.7 umol/kg) OC from Compound KC-14 (5.7 umol/kg) OC from Compound KC-15 (5.7 umol/kg) OC from Compound KC-16 (5.8 umol/kg) OC from Compound KC-17 (5.8 umol/kg)

0.1

Time, hr

Figure 5B

1000 -o- OC from Compound KC-17 (5.8 umol/kg) -a- OC from Oxycodone (5.7umol/kg) -0- OC from 20 mg OxyContin Tablet

-H OC from Compound KC-3 (5.7 umol/kg)

Time, hr U.S. Patent Apr. 1, 2014 Sheet 7 of 26 US 8,685,916 B2

Figure 6A -o- OC from Compound KC-12 (6.5umol/kg) 100 -- OC from Compound KC-12 (29 umol/kg)

- 10 E o C g O 1 C O C X 9. O. 1

O.O1 O 6 12 18 24 Time, hr Figure 6B or OC from Compound KC-17 (6 umol/kg) 100 -- OC from Compound KC-17 (28 umol/kg) - OC from Compound KC-17 (60 umol/kg)

1O S g O 1 C O ce X 9. O.1

O.O1 O 6 12 18 24 Time, hr U.S. Patent Apr. 1, 2014 Sheet 8 of 26 US 8,685,916 B2

Figure 7A

-O OC from Compound KC-12 (6.5umol/kg) H OC from Compound KC-12 (6.5umol/kg)( ) + Compound 109 (0.2 umol/kg) the OC from Compound KC-12 (6.5umol/kg) + Compound 109 (0.9 umol/kg) -0. OC from Compound KC-12 (6.5umol/kg)( ) + Compound 109 (1.9 umol/kg) 4

3

2

Time, hr Figure 7B -o- OC from Compound KC-17 (6 umol/kg) H. OC from Compound KC-17 (6 umol/kg) + Compound 109 (0.2 umol/kg) -A- OC from Compound KC-17 (6 umol/kg) + Compound 109 (0.9 umol/kg)

-0- OC from Compound KC-17 (6 umol/kg) + Compound 109 (1.9 umol/kg) U.S. Patent Apr. 1, 2014 Sheet 9 of 26 US 8,685,916 B2

Figure 7C -o- OC from Compound KC-17 (60 umol/kg -H OC from Compound KC-17 (60 umol/kg + Compound 109 (1.9 umol/kg) -- OC from Compound KC-17 (60 umol/kg + Compound 109 (9.3 umol/kg) -0- OC from Compound KC-17 (60 umol/kg + Compound 109 (18.5umol/kg)

40 23 OO

1 O U.S. Patent Apr. 1, 2014 Sheet 10 of 26 US 8,685,916 B2

Figure 8 -O- Tapentadol from Compound TP-5 (34 umol/kg) H. Tapentadol from Compound TP-5 (34 umol/kg) + Compound 109 (55umol/kg)

Time, hr U.S. Patent Apr. 1, 2014 Sheet 11 of 26 US 8,685,916 B2

Figure9

-- Hydrocodone from Compound KC-31 (28 umol/kg) 'N. -o- Hydrocodone from Compound KC-31 (12 umol/kg) 's. -- Hydrocodone from Compound KC-31 (6 umol/kg) - Hydrocodone from HC (30 umol/kg)

O 1 2 3 4 5 66 9 12 15 18 21 24 Time, hr U.S. Patent Apr. 1, 2014 Sheet 12 of 26 US 8,685,916 B2

Figure 10

6 -O Hydrocodone from HC (30 umol/kg) -- Hydrocodone from Compound KC-32 (28 umol/kg )

- Hydrocodone from Compound KC-35 (24 umol/kg ) -0- Hydrocodone from Compound KC-36 (27 umol/kg) s --- Hydrocodone from Compound KC-37 (28 umol/kg ) 4.

3

Time, hr U.S. Patent Apr. 1, 2014 Sheet 13 of 26 US 8,685,916 B2

Figure 11

-o- Hydrocodone from HC (30 umol/kg) -v- Hydrocodone from Compound KC-38 (28 umol/kg) -e Hydrocodone from Compound KC-39 (28 umol/kg)

O 2 4 6 8 Time, hr U.S. Patent Apr. 1, 2014 Sheet 14 of 26 US 8,685,916 B2

Figure 12

6 -O Hydrocodone from HC (30 umol/kg) -- Hydrocodone from Compound KC-40 (28 mol/kg) 5 de Hydrocodone from Compound KC-47 (28 umol/kg)

-X- Hydrocodone from Compound KC-50 (28 umol/kg)

Time, hr U.S. Patent Apr. 1, 2014 Sheet 15 of 26 US 8,685,916 B2

Figure 13A

1O -o- Hydrocodone (HC) from Compound KC-40 (6 umol/kg) -- HC from Compound KC-40 (6 umol/kg) + Compound 109 (0.2 umol/kg) 8 -)- HC from Compound KC-40 (6 umol/kg) + Compound 109 (0.44 umol/kg) -- HC from Compound KC-40 (6 umol/kg) + Compound 109 (0.9Lumol/kg) S. E 6 -- HC from Compound KC-40 (6 umol/kg) + Compound 109 (1.7 umol/kg)

O O o 9 e E.

Time, hr U.S. Patent Apr. 1, 2014 Sheet 16 of 26 US 8,685,916 B2

Figure 13B -H Hydrocodone from HC (30pumol/kg normalized to 65umol/kg) -- HC from Compound KC-40 (62 umol/kg) + Compound 109 (1.7 umol/kg) -)- HC from Compound KC-40 (62 umol/kg) + Compound 109 (4.4 umol/kg) -- HC from Compound KC-40 (62 umol/kg) + Compound 109 (9.0 umol/kg) -- HC from Compound KC-40 (62 umol/kg)) + Compound 109 (17.4 umol/kg)

Time, hr U.S. Patent Apr. 1, 2014 Sheet 17 of 26 US 8,685,916 B2

Figure 13C -o- Hydrocodone (HC) from Compound KC-50 (6 umol/kg) 12 sh HC from Compound KC-50 (6 umol/kg) + Compound 109 (0.2 umol/kg) -x- HC from Compound KC-50 (6 umol/kg) + Compound 109 (0.44 umol/kg) -- HC from Compound KC-50 (6 umol/kg) + Compound 109 (0.9 umol/kg) 1O -- HC from Compound KC-50 (6 umol/kg) + Compound 109 (1.7 umol/kg)

- S 8 o d Os5 6 o 9 O i 4

Time, hr U.S. Patent Apr. 1, 2014 Sheet 18 of 26 US 8,685,916 B2

Figure 13D

es Hydrocodone from HC (30 umol/kg normalized to 67 umol/kg) HC from Compound KC-50 (64 umol/kg) + Compound 109 (1.7 umol/kg) 12 ex HC from Compound KC-50 (64 umol/kg) + Compound 109 (4.4 pmol/kg) es HC from Compound KC-50 (64 umol/kg) + Compound 109 (9.0 pumol/kg)

Time, hr U.S. Patent Apr. 1, 2014 Sheet 19 of 26 US 8,685,916 B2

Figure 14A

80 -- Hydrocodone from HC (0.5umol/kg) -o-, HC from Compound KC-40 (0.1 umol/kg) -- HC from Compound KC-40 (0.5umol/kg) 60 -)- HC from Compound KC-40 (2.0 umol/kg)

4.O

Time, hr U.S. Patent Apr. 1, 2014 Sheet 20 of 26 US 8,685,916 B2

Figure 14B

15 -- Hydrocodone from HC (0.5umol/kg) -o-, HC from Compound KC-40 (0.5umol/kg) + Compound 109 (0.14 mol/kg)

1 O

5

Time, hr

Figure 14C

150

-H Hydrocodone from HC (0.5umol/kg normalized to 2 umol/kg) -o-, HC from Compound KC-40 (2.0 umol/kg) + Compound 109 (0.56 umol/kg)

Time, hr U.S. Patent Apr. 1, 2014 Sheet 21 of 26 US 8,685,916 B2

Figure 14D -H Hydrocodone from HC (0.5 pmol/kg normalized to 5umol/kg) 150 or HC from Compound KC-40 (4.7umol/kg) + Compound 109 (1.4 umol/kg)

i

Time, hr U.S. Patent Apr. 1, 2014 Sheet 22 of 26 US 8,685,916 B2

Figure 15A

-H Hydrocodone from HC (0.5 pmol/kg) 60 -o-, HC from Compound KC-50 (0.1 p.mol/kg) -- HC from Compound KC-50 (0.5umol/kg) -- HC from Compound KC-50 (2 umol/kg)

4O

20

Time, hr U.S. Patent Apr. 1, 2014 Sheet 23 of 26 US 8,685,916 B2

Figure 15B

15 -H Hydrocodone from HC (0.5umol/kg) - O- HC from Compound KC-50 (0.5umol/kg) the HC from Compound KC-50 (0.5umol/kg) + Compound 109 (0.14 umol/kg) -)- HC from Compound KC-50 (0.5umol/kg) + Compound 109 (0.28 umol/kg) E

ab s O 9 ce E.

Time, hr Figure 15C -- Hydrocodone from HC (0.5umol/kg normalized to 5 umol/kg) 150 HC from Compound KC-50 (5umol/kg) + Compound 109 (0.14 umol/kg) -x- HC from Compound KC-50 (5umol/kg) + Compound 109 (0.7 umol/kg)

100

50

Time, hr U.S. Patent Apr. 1, 2014 Sheet 24 of 26 US 8,685,916 B2

Figure 16 -H Oxycodone from OC (28 umol/kg) 16 -- OC from Compound KC-55 (6 umol/kg) W W -o- OC from Compound KC-55 (28 umol/kg) W

Time, hr U.S. Patent Apr. 1, 2014 Sheet 25 of 26 US 8,685,916 B2

Figure 17A

20 -o- OC from Compound KC-55 (6 mol/kg) -- OC from Compound KC-55 (6 umol/kg)+ Compound 109 (0.9 umol/kg)

15

10 U.S. Patent Apr. 1, 2014 Sheet 26 of 26 US 8,685,916 B2

Figure 17B H. Oxycodone (28 umol/kg normalized to 36 unol/kg) -o- OC from Compound KC-55 (36 mol/kg) + Compound 109 (5umol/kg) 20 US 8,685,916 B2 1. 2 OPOD PRODRUGS WITH HETEROCYCLC hydrogenatom of the phenolic hydroxyl group is replaced by LINKERS a covalent bond to —C(O) N(A ring)-Y. (CRR) NH CO) CH(R) N(R) C(O) CH(R) CROSS REFERENCE TO RELATED N(R) R', and a residue of an amide-containing opioid, APPLICATIONS wherein - C(O) N(A ring)-Y-(CRR). NH C (O) CH(R) N(R) C(O) CH(R) N(R) R7 is This application claims the benefit of U.S. Provisional connected to the amide-containing opioid through the oxygen Application No. 61/451,010 filed Mar. 9, 2011 and U.S. Pro of the amide group, wherein the amide group is converted to visional Application No. 61/583,516 filed Jan.5, 2012, which an amide enol or an imine tautomer, 10 the A ring is a heterocyclic 5 to 12-membered ring: are hereby incorporated by reference in their entireties. each Y is independently selected from alkyl, substituted alkyl, alkenyl, Substituted alkenyl, alkynyl, Substituted alky INTRODUCTION nyl, aryl, Substituted aryl, acyl, Substituted acyl, carboxyl, Opioids are susceptible to misuse, abuse, or overdose. Use alkoxycarbonyl, Substituted alkoxycarbonyl, aminoacyl, Sub 15 stituted aminoacyl, amino, Substituted amino, acylamino, of and access to these drugs therefore needs to be controlled. Substituted acylamino, and cyano; The control of access to the drugs is expensive to administer c is a number from Zero to 3: and can result in denial of treatment for patients that are not each R' is independently selected from hydrogen, alkyl, able to present themselves for dosing. For example, patients substituted alkyl, alkenyl, substituted alkenyl, alkynyl, sub Suffering from acute pain may be denied treatment with an stituted alkynyl, aryl, Substituted aryl, acyl, Substituted acyl, opioid unless they have been admitted to a hospital. Further carboxyl, alkoxycarbonyl, Substituted alkoxycarbonyl, ami more, control of use is often ineffective, leading to Substantial noacyl, Substituted aminoacyl, amino, Substituted amino, morbidity and deleterious Social consequences. acylamino, Substituted acylamino, and cyano; SUMMARY each R is independently selected from hydrogen, alkyl, 25 substituted alkyl, alkenyl, substituted alkenyl, alkynyl, sub stituted alkynyl, aryl, Substituted aryl, acyl, Substituted acyl, This disclosure concerns a prodrug of an opioid that pro carboxyl, alkoxycarbonyl, Substituted alkoxycarbonyl, ami vides controlled release of the opioid. Such a prodrug com prises an opioid covalently attached to a promoiety. The pro noacyl, Substituted aminoacyl, amino, Substituted amino, moiety comprises an enzyme-cleavable moiety and a acylamino, Substituted acylamino, and cyano; or 30 R" and R together with the carbon to which they are cyclizable spacer leaving group Such that the opioid prodrug attached can form a cycloalkyl or Substituted cycloalkyl provides controlled release of opioid via enzyme cleavage group, or two R' or R groups on adjacent carbon atoms, followed by intramolecular cyclization. The enzyme-medi together with the carbonatoms to which they are attached, can ated release of the opioid can occur in the gastrointestinal form a cycloalkyl or Substituted cycloalkyl group; tract upon oral administration of the corresponding prodrug. a is an integer from one to 8: Thus, prodrugs of the disclosure provide efficient delivery of provided that whena is one, the A ring is a heterocyclic 6 to opioid when ingested. 12-membered ring; and when the A ring is a heterocyclic The present disclosure also provides a composition, Such 5-membered ring, then a is an integer from 2 to 8; as a pharmaceutical composition, that comprises an opioid each R is independently hydrogen, alkyl, substituted prodrug of the embodiments. Such a composition can option 40 alkyl, aryl or substituted aryl; ally provide an inhibitor that interacts with the enzyme that R is selected from hydrogen, alkyl, substituted alkyl, aryl, mediates the controlled release of opioid from the prodrug so substituted aryl, arylalkyl, substituted arylalkyl, heteroalkyl, as to attenuate enzymatic cleavage of the prodrug. The dis substituted heteroalkyl, heteroaryl, substituted heteroaryl, closure provides for the enzyme being a gastrointestinal (GI) heteroarylalkyl, and substituted heteroarylalkyl: enzyme, such as trypsin. Also provided are methods of use, 45 each R is independently selected from hydrogen, alkyl, Such as a method of providing patients with controlled release substituted alkyl, aryl, substituted aryl, arylalkyl, substituted of opioid using an opioid prodrug of the embodiments. arylalkyl, heteroalkyl, substituted heteroalkyl, heteroaryl, The embodiments include an opioid prodrug that is a com substituted heteroaryl, heteroarylalkyl, and substituted het pound of formula I: eroarylalkyl: 50 b is a number from Zero to 100; and R’ is selected from hydrogen, alkyl, substituted alkyl, acyl, Substituted acyl, alkoxycarbonyl, Substituted alkoxycarbo O R1 R2 O R3 R6 nyl, aryl, Substituted aryl, arylalkyl, and Substituted arylalkyl; or a salt, hydrate or solvate thereof. 55 Certain embodiments provide controlled release of ketone X N a N N containing opioids. More particularly, the embodiments ls A 1.R5 O k r relate to a prodrug of a ketone-containing opioid that provides controlled release of the opioid. Such a prodrug comprises a (Y), ketone-containing opioid covalently attached to a promoiety 60 through the enolic oxygen atom of the ketone-containing wherein opioid. The promoiety comprises an enzyme-cleavable moi X is selected from a residue of a ketone-containing opioid, ety and a cyclizable spacer leaving group Such that the ketone wherein the hydrogen atom of the corresponding hydroxyl modified opioid prodrug provides controlled release of opioid group of the enolic tautomer of the ketone is replaced by a via enzyme cleavage followed by intramolecular cyclization. covalent bond to —C(O) NI (A ring)-Y. (CRR), 65 Ketone-modified opioid prodrugs of the disclosure provide NH C(O) CH(R) N(R) C(O) CH(R) efficient delivery of opioid when ingested. The present dis N(R) R'; a residue of a phenolic opioid, wherein the closure also provides a composition, such as a pharmaceutical US 8,685,916 B2 3 4 composition, that comprises a ketone-modified opioid pro R’ is selected from hydrogen, alkyl, substituted alkyl, acyl, drug of the embodiments. Also provided are methods of use, Substituted acyl, alkoxycarbonyl, Substituted alkoxycarbo Such as a method of providing patients with controlled release nyl, aryl, Substituted aryl, arylalkyl, and Substituted arylalkyl; of ketone-containing opioid using a ketone-modified opioid or a salt, hydrate or solvate thereof. prodrug of the embodiments. Certain embodiments provide controlled release of phe The embodiments include a ketone-modified opioid pro nolic opioids. More particularly, the embodiments relate to a drug that is a compound of formula II: prodrug of a phenolic opioid that provides controlled release of the opioid. Such a prodrug comprises a phenolic opioid covalently attached to a promoiety through the phenolic oxy (II) 10 gen atom of the phenolic opioid. The promoiety comprises an enzyme-cleavable moiety and a cyclizable spacer leaving O R1 R2 O R3 R6 group Such that the phenolic opioid prodrug provides con trolled release of opioid via enzyme cleavage followed by X N a N N 15 intramolecular cyclization. Phenolic opioid prodrugs of the l A 1.R5 O r disclosure provide efficient delivery of opioid when ingested. The present disclosure also provides a composition, Such as a (Y), pharmaceutical composition, that comprises a phenolic opioid prodrug of the embodiments. Also provided are meth wherein ods of use, Such as a method of providing patients with X represents a residue of a ketone-containing opioid, controlled release of phenolic opioid using a phenolic opioid wherein the hydrogen atom of the corresponding hydroxyl prodrug of the embodiments. group of the enolic tautomer of the ketone is replaced by a The embodiments include a phenolic opioid prodrug that is covalent bond to —C(O) NI (A ring)-Y. (CRR), a compound of formula VI: NH C(O) CH(R) N(R) C(O) CH(R) 25 N(R) R7: the A ring is a heterocyclic 5 to 12-membered ring; each Y is independently selected from alkyl, substituted O R1 R2 O R3 R6 alkyl, alkenyl, Substituted alkenyl, alkynyl, Substituted alky nyl, aryl, Substituted aryl, acyl, Substituted acyl, carboxyl, 30 alkoxycarbonyl. Substituted alkoxycarbonyl, aminoacyl, Sub X N a 'N N stituted aminoacyl, amino, substituted amino, acylamino, ls A. h 1.RS O Substituted acylamino, and cyano; c is a number from Zero to 3: (Y), each R" is independently selected from hydrogen, alkyl, 35 substituted alkyl, alkenyl, substituted alkenyl, alkynyl, sub wherein stituted alkynyl, aryl, Substituted aryl, acyl, Substituted acyl, X represents a residue of a phenolic opioid, wherein the carboxyl, alkoxycarbonyl, Substituted alkoxycarbonyl, ami hydrogenatom of the phenolic hydroxyl group is replaced by noacyl, Substituted aminoacyl, amino, Substituted amino, a covalent bond to —C(O) N (A ring)-Y. (CRR), acylamino, Substituted acylamino, and cyano; 40 NH CO) CH(R) N(R) C(O) CH(R) each R is independently selected from hydrogen, alkyl, N(R) R7: substituted alkyl, alkenyl, substituted alkenyl, alkynyl, sub the A ring is a heterocyclic 5 to 12-membered ring: stituted alkynyl, aryl, Substituted aryl, acyl, Substituted acyl, each Y is independently selected from alkyl, substituted carboxyl, alkoxycarbonyl, Substituted alkoxycarbonyl, ami alkyl, alkenyl, Substituted alkenyl, alkynyl, Substituted alky noacyl, Substituted aminoacyl, amino, Substituted amino, 45 nyl, aryl, Substituted aryl, acyl, Substituted acyl, carboxyl, acylamino, Substituted acylamino, and cyano; or alkoxycarbonyl, Substituted alkoxycarbonyl, aminoacyl, Sub R" and R together with the carbon to which they are stituted aminoacyl, amino, Substituted amino, acylamino, attached can form a cycloalkyl or Substituted cycloalkyl Substituted acylamino, and cyano; group, or two R' or R groups on adjacent carbon atoms, c is a number from Zero to 3: together with the carbonatoms to which they are attached, can 50 each R' is independently selected from hydrogen, alkyl, form a cycloalkyl or Substituted cycloalkyl group; substituted alkyl, alkenyl, substituted alkenyl, alkynyl, sub a is an integer from one to 8: stituted alkynyl, aryl, Substituted aryl, acyl, Substituted acyl, provided that whena is one, the Aring is a heterocyclic 6 to carboxyl, alkoxycarbonyl, Substituted alkoxycarbonyl, ami 12-membered ring; and when the A ring is a heterocyclic noacyl, Substituted aminoacyl, amino, Substituted amino, 5-membered ring, then a is an integer from 2 to 8: 55 acylamino, Substituted acylamino, and cyano; each R is independently hydrogen, alkyl, substituted each R is independently selected from hydrogen, alkyl, alkyl, aryl or substituted aryl; substituted alkyl, alkenyl, substituted alkenyl, alkynyl, sub R is selected from hydrogen, alkyl, substituted alkyl, aryl, stituted alkynyl, aryl, Substituted aryl, acyl, Substituted acyl, substituted aryl, arylalkyl, substituted arylalkyl, heteroalkyl, carboxyl, alkoxycarbonyl, Substituted alkoxycarbonyl, ami substituted heteroalkyl, heteroaryl, substituted heteroaryl, 60 noacyl, Substituted aminoacyl, amino, Substituted amino, heteroarylalkyl, and substituted heteroarylalkyl; acylamino, Substituted acylamino, and cyano; or each R is independently selected from hydrogen, alkyl, R" and R together with the carbon to which they are substituted alkyl, aryl, substituted aryl, arylalkyl, substituted attached can form a cycloalkyl or Substituted cycloalkyl arylalkyl, heteroalkyl, substituted heteroalkyl, heteroaryl, group, or two R' or R groups on adjacent carbon atoms, substituted heteroaryl, heteroarylalkyl, and substituted het 65 together with the carbonatoms to which they are attached, can eroarylalkyl: form a cycloalkyl or Substituted cycloalkyl group; b is a number from Zero to 100; and a is an integer from one to 8: US 8,685,916 B2 5 6 provided that whena is one, the Aring is a heterocyclic 6 to each R' is independently selected from hydrogen, alkyl, 12-membered ring; and when the A ring is a heterocyclic substituted alkyl, alkenyl, substituted alkenyl, alkynyl, sub 5-membered ring, then a is an integer from 2 to 8: stituted alkynyl, aryl, Substituted aryl, acyl, Substituted acyl, each R is independently hydrogen, alkyl, substituted carboxyl, alkoxycarbonyl, Substituted alkoxycarbonyl, ami alkyl, aryl or substituted aryl; 5 noacyl, Substituted aminoacyl, amino, Substituted amino, R is selected from hydrogen, alkyl, substituted alkyl, aryl, acylamino, Substituted acylamino, and cyano; substituted aryl, arylalkyl, substituted arylalkyl, heteroalkyl, each R is independently selected from hydrogen, alkyl, substituted heteroalkyl, heteroaryl, substituted heteroaryl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, sub heteroarylalkyl, and substituted heteroarylalkyl; stituted alkynyl, aryl, Substituted aryl, acyl, Substituted acyl, each R is independently selected from hydrogen, alkyl, 10 carboxyl, alkoxycarbonyl, Substituted alkoxycarbonyl, ami substituted alkyl, aryl, substituted aryl, arylalkyl, substituted noacyl, Substituted aminoacyl, amino, Substituted amino, arylalkyl, heteroalkyl, substituted heteroalkyl, heteroaryl, acylamino, Substituted acylamino, and cyano; or substituted heteroaryl, heteroarylalkyl, and substituted het R" and R together with the carbon to which they are eroarylalkyl: attached can form a cycloalkyl or Substituted cycloalkyl b is a number from Zero to 100: 15 R’ is selected from hydrogen, alkyl, substituted alkyl, acyl, group, or two R' or R groups on adjacent carbon atoms, Substituted acyl, alkoxycarbonyl, Substituted alkoxycarbo together with the carbonatoms to which they are attached, can nyl, aryl, Substituted aryl, arylalkyl, and Substituted arylalkyl; form a cycloalkyl or Substituted cycloalkyl group; or a salt, hydrate or solvate thereof. a is an integer from one to 8: Certain embodiments provide controlled release of amide provided that whena is one, the A ring is a heterocyclic 6 to containing opioids. More particularly, the embodiments 12-membered ring; and when the A ring is a heterocyclic relate to a prodrug of an amide-containing opioid that pro 5-membered ring, then a is an integer from 2 to 8; vides controlled release of the opioid. Such a prodrug com each R is independently hydrogen, alkyl, substituted prises an amide-containing opioid covalently attached to a alkyl, aryl or substituted aryl; promoiety through the enolic oxygen atom of the amide enol 25 R is selected from hydrogen, alkyl, substituted alkyl, aryl, moiety or through the oxygen of the imine tautomer of the substituted aryl, arylalkyl, substituted arylalkyl, heteroalkyl, amide-containing opioid. The promoiety comprises an substituted heteroalkyl, heteroaryl, substituted heteroaryl, enzyme-cleavable moiety and a cyclizable spacer leaving heteroarylalkyl, and substituted heteroarylalkyl: group Such that the amide-modified opioid prodrug provides each R is independently selected from hydrogen, alkyl, controlled release of opioid via enzyme cleavage followed by 30 substituted alkyl, aryl, substituted aryl, arylalkyl, substituted intramolecular cyclization. Amide-modified opioid prodrugs arylalkyl, heteroalkyl, substituted heteroalkyl, heteroaryl, of the disclosure provide efficient delivery of opioid when substituted heteroaryl, heteroarylalkyl, and substituted het ingested. The present disclosure also provides a composition, eroarylalkyl: Such as a pharmaceutical composition, that comprises an b is a number from Zero to 100: amide-modified opioid prodrug of the embodiments. Also 35 R’ is selected from hydrogen, alkyl, substituted alkyl, acyl, provided are methods of use, such as a method of providing Substituted acyl, alkoxycarbonyl, Substituted alkoxycarbo patients with controlled release of amide-containing opioid nyl, aryl, Substituted aryl, arylalkyl, and Substituted arylalkyl; using an amide-modified opioid prodrug of the embodiments. or a salt, hydrate or solvate thereof. The embodiments include an amide containing opioid pro drug that is a compound of formula X: 40 BRIEF DESCRIPTION OF THE FIGURES FIG. 1 is a schematic representing the effect of increasing the level of a GI enzyme inhibitor (“inhibitor. X axis) on a (X) PK parameter (e.g., drug Cmax) (Yaxis) for a fixed dose of l R1 R2 O R3 R6 45 prodrug. The effect of inhibitor upon a prodrug PK parameter R7 can range from undetectable, to moderate, to complete inhi X N d N bition (i.e., no detectable drug release). A N FIG. 2 provides schematics of drug concentration in H R5 O R3 plasma (Yaxis) over time (X axis). Panel A is a schematic of 50 a pharmacokinetic (PK) profile following ingestion of pro (Y), drug with a GI enzyme inhibitor (dashed line) where the drug Cmax is modified relative to that of prodrug without inhibitor wherein (solid line). Panel B is a schematic of a PK profile following X represents a residue of an amide-containing opioid, ingestion of prodrug with inhibitor (dashed line) where drug wherein –C(O) NLA ring)-Y-(CRR). NH C 55 Cmax and drug Tmax are modified relative to that of prodrug (O) CH(R) N(R)-C(O)-CH(R) N(R) R7 is without inhibitor (solid line). Panel C is a schematic of a PK connected to the amide-containing opioid through the oxygen profile following ingestion of prodrug with inhibitor (dashed of the amide group, wherein the amide group is converted to line) where drug Tmax is modified relative to that of prodrug an amide enol or an imine tautomer, without inhibitor (solid line). the A ring is a heterocyclic 5 to 12-membered ring; 60 FIG. 3 provides schematics representing differential con each Y is independently selected from alkyl, substituted centration-dose PK profiles that can result from the dosing of alkyl, alkenyl, Substituted alkenyl, alkynyl, Substituted alky multiples of a dose unit (X axis) of the present disclosure. nyl, aryl, Substituted aryl, acyl, Substituted acyl, carboxyl, Different PK profiles (as exemplified herein for a representa alkoxycarbonyl. Substituted alkoxycarbonyl, aminoacyl, Sub tive PK parameter, drug Cmax (Yaxis)) can be provided by stituted aminoacyl, amino, Substituted amino, acylamino, 65 adjusting the relative amount of prodrug and GI enzyme Substituted acylamino, and cyano; inhibitor contained in a single dose unit or by using a different c is a number from Zero to 3: prodrug or inhibitor in the dose unit. US 8,685,916 B2 7 8 FIG. 4 compares mean plasma concentrations over time of increasing amounts of co-dosed trypsin inhibitor Compound oxycodone release following PO administration to rats of 109 to hydrocodone values expected from a normalized several ketone-modified opioid prodrugs of the embodi hydrocodone dose. mentS. FIG. 14A compares mean plasma concentrations overtime FIG. 5A compares mean plasma concentrations over time 5 of hydrocodone following PO administration to dogs of of oxycodone release following PO administration to dogs of hydrocodone and PO administration to dogs of increasing several ketone-modified opioid prodrugs of the embodi amounts of prodrug Compound KC-40. FIG. 14B, FIGS. 14C ments. FIG. 5B compares mean plasma concentrations over and 14D each compares mean plasma concentrations over time of oxycodone release following PO administration to time of hydrocodone following PO administration to dogs of dogs of ketone-modified opioid prodrug Compound KC-17 10 respectively, 1, 4 or 10 dose units comprising prodrug Com of the embodiments, oxycodone prodrug Compound KC-3, pound KC-40 and trypsin inhibitor Compound 109 to plasma OxyContin R tablets, or oxycodone HC1. concentrations of hydrocodone following PO administered to FIG. 6A compares mean plasma concentrations over time dogs of 1 dose equivalent of hydrocodone or predicted con of oxycodone release following PO administration to rats of 15 centrations for 4 or 10 dose equivalents of hydrocodone, increasing doses of ketone-modified opioid prodrug Com respectively. pound KC-12. FIG. 6B compares mean plasma concentra FIG. 15A compares mean plasma concentrations overtime tions over time of oxycodone release following PO adminis of hydrocodone following PO administration to dogs of tration to rats of increasing doses of ketone-modified opioid hydrocodone and PO administration to dogs of increasing prodrug Compound KC-17. amounts of prodrug Compound KC-50. FIG. 15B and FIG. FIG. 7A compares mean plasma concentrations over time 15C compare mean plasma concentrations over time of of oxycodone release following PO administration to rats of hydrocodone following PO administration to dogs of hydro ketone-modified opioid prodrug Compound KC-12 co-dosed codone to plasma concentrations over time of hydrocodone with increasing amounts of trypsin inhibitor Compound 109. following PO administration to dogs of the indicated doses of FIG. 7B and FIG. 7C compare mean plasma concentrations 25 prodrug Compound KC-50 with or without trypsin inhibitor over time of oxycodone release following PO administration Compound 109. to rats of two doses of ketone-modified opioid prodrug Com FIG. 16 compares mean plasma concentrations over time pound KC-17, each co-dosed with increasing amounts of of oxycodone release following PO administration to rats of trypsin inhibitor Compound 109. oxycodone prodrug Compound KC-55 or oxycodone. FIG. 8 compares mean plasma concentrations over time of 30 FIG. 17A provides oxycodone exposure results for rats tapentadol release following PO administration to rats of orally administered prodrug Compound KC-55 alone or co phenolic opioid prodrug Compound TP-5 in the absence or dosed with trypsin inhibitor Compound 109. FIG. 17B pro presence of a co-dose of trypsin inhibitor Compound 109. vides oxycodone exposure results for rats orally administered FIG. 9 compares mean plasma concentrations over time of prodrug Compound KC-55 alone or co-dosed with trypsin hydrocodone release following PO administration to rats of 35 inhibitor Compound 109. increasing doses of ketone-modified opioid prodrug Com pound KC-31. TERMS FIG. 10 compares mean plasma concentrations over time ofhydrocodone release following PO administration to rats of 40 The following terms have the following meaning unless hydrocodone and PO administration to rats of hydrocodone otherwise indicated. Any undefined terms have their art rec prodrugs Compound KC-32, Compound KC-35, Compound ognized meanings. KC-36, and Compound KC-37. Alkyl by itself or as part of another substituent refers to FIG. 11 compares mean plasma concentrations over time a saturated branched or straight-chain monovalent hydrocar ofhydrocodone release following PO administration to rats of 45 bon radical derived by the removal of one hydrogen atom hydrocodone and PO administration to rats of hydrocodone from a single carbon atom of a parent alkane. Typical alkyl prodrugs Compound KC-38 and Compound KC-39. groups include, but are not limited to, methyl, ethyl, propyls FIG. 12 compares mean plasma concentrations over time Such as propan-1-yl or propan-2-yl; and butyls such as butan ofhydrocodone release following PO administration to rats of 1-yl, butan-2-yl, 2-methyl-propan-1-yl or 2-methyl-propan hydrocodone and PO administration to rats of hydrocodone 50 2-yl. In some embodiments, an alkyl group comprises from 1 prodrugs Compound KC-40, Compound KC-47, and Com to 20 carbon atoms. In other embodiments, an alkyl group pound KC-50. comprises from 1 to 10 carbon atoms. In still other embodi FIG. 13A compares mean plasma concentrations over time ments, an alkyl group comprises from 1 to 6 carbon atoms, ofhydrocodone release following PO administration to rats of Such as from 1 to 4 carbon atoms. prodrug Compound KC-40 with increasing amounts of co 55 “Alkanyl by itselforas part of another substituent refers to dosed trypsin inhibitor Compound 109. FIG. 13B compares a Saturated branched, straight-chain or cyclic alkyl radical mean plasma concentrations over time of hydrocodone derived by the removal of one hydrogen atom from a single release following PO administration to rats of prodrug Com carbonatom of an alkane. Typical alkanyl groups include, but pound KC-40 with increasing amounts of co-dosed trypsin are not limited to, methanyl; ethanyl; propanyls such as pro inhibitor Compound 109 to hydrocodone values expected 60 pan-1-yl, propan-2-yl (isopropyl), cyclopropan-1-yl, etc.; from a normalized hydrocodone dose. FIG. 13C compares the butanyls such as butan-1-yl, butan-2-yl (sec-butyl), 2-methyl mean plasma concentrations over time of hydrocodone propan-1-yl (isobutyl), 2-methyl-propan-2-yl (t-butyl), release following PO administration to rats of prodrug Com cyclobutan-1-yl, etc.; and the like. pound KC-50 with increasing amounts of co-dosed trypsin Alkylene' refers to a branched or unbranched saturated inhibitor Compound 109. FIG. 13D compares mean plasma 65 hydrocarbon chain, usually having from 1 to 40 carbonatoms, concentrations over time of hydrocodone release following more usually 1 to 10 carbonatoms and even more usually 1 to PO administration to rats of prodrug Compound KC-50 with 6 carbon atoms. This term is exemplified by groups such as US 8,685,916 B2 9 10 methylene (—CH2—), ethylene (-CH2CH2—), the propy cycloalkyl, cycloalkenyl, Substituted cycloalkenyl, aryl, Sub lene isomers (e.g., —CH2CHCH - and —CH(CH) stituted aryl, heteroaryl, substituted heteroaryl, heterocyclic, CH ) and the like. and substituted heterocyclic are as defined herein. Alkenyl by itselforas part of another substituent refers to “Alkoxy” by itselfor as part of another substituent refers to an unsaturated branched, straight-chain orcyclic alkyl radical a radical —OR' where R represents an alkyl or cycloalkyl having at least one carbon-carbon double bond derived by the group as defined herein. Representative examples include, removal of one hydrogen atom from a single carbon atom of but are not limited to, methoxy, ethoxy, propoxy, butoxy, an alkene. The group may be in either the cis or trans confor cyclohexyloxy and the like. mation about the double bond(s). Typical alkenyl groups “Alkoxycarbonyl by itselfor as part of another substituent include, but are not limited to, ethenyl; propenyls such as 10 refers to a radical—C(O)OR where R' represents an alkyl prop-1-en-1-yl, prop-1-en-2-yl, prop-2-en-1-yl (allyl), prop or cycloalkyl group as defined herein. Representative 2-en-2-yl, cycloprop-1-en-1-yl, cycloprop-2-en-1-yl; bute examples include, but are not limited to, methoxycarbonyl, nyls such as but-1-en-1-yl, but-1-en-2-yl, 2-methyl-prop-1- ethoxycarbonyl, propoxycarbonyl, butoxycarbonyl, cyclo en-1-yl, but-2-en-1-yl, but-2-en-1-yl, but-2-en-2-yl, buta-1, hexyloxycarbonyl and the like. 3-dien-1-yl, buta-1,3-dien-2-yl, cyclobut-1-en-1-yl, 15 Aryl by itself or as part of another substituent refers to a cyclobut-1-en-3-yl, cyclobuta-1,3-dien-1-yl, etc.; and the monovalent aromatic hydrocarbon radical derived by the like. removal of one hydrogen atom from a single carbon atom of Alkynyl by itselforas part of another substituent refers to an aromatic ring system. Typical aryl groups include, but are an unsaturated branched, straight-chain orcyclic alkyl radical not limited to, groups derived from aceanthrylene, acenaph having at least one carbon-carbon triple bond derived by the thylene, acephenanthrylene, anthracene, aZulene, benzene, removal of one hydrogen atom from a single carbon atom of chrysene, coronene, fluoranthene, fluorene, hexacene, an alkyne. Typical alkynyl groups include, but are not limited hexaphene, hexylene, as-indacene, S-indacene, indane, to, ethynyl; propynyls such as prop-1-yn-1-yl, prop-2-yn-1- indene, naphthalene, octacene, octaphene, octalene, ovalene, yl, etc.; butynyls such as but-1-yn-1-yl, but-1-yn-3-yl, but-3- penta-2,4-diene, pentacene, pentalene, pentaphene, perylene, yn-1-yl, etc.; and the like. 25 phenalene, phenanthrene, picene, pleiadene, pyrene, pyran Acyl by itself or as part of another substituent refers to a threne, rubicene, triphenylene, trinaphthalene and the like. In radical—C(O)R', where R' is hydrogen, alkyl, cycloalkyl, certain embodiments, an aryl group comprises from 6 to 20 cycloheteroalkyl, aryl, arylalkyl, heteroalkyl, heteroaryl, het carbon atoms. In certain embodiments, an aryl group com eroarylalkyl as defined herein and substituted versions prises from 6 to 12 carbon atoms. Examples of an aryl group thereof. Representative examples include, but are not limited 30 are phenyl and naphthyl. to formyl, acetyl, cyclohexylcarbonyl, cyclohexylmethylcar Arylalkyl by itselfor as part of another substituent refers bonyl, benzoyl, benzylcarbonyl, piperonyl, succinyl, and to an acyclic alkyl radical in which one of the hydrogenatoms malonyl, and the like. bonded to a carbon atom, typically a terminal or sp carbon “Acylamino” refers to the groups —NRC(O)alkyl, atom, is replaced with an aryl group. Typical arylalkyl groups - NRC(O)substituted alkyl, NRC(O)cycloalkyl, 35 include, but are not limited to, benzyl, 2-phenylethan-1-yl, -NRC(O)substituted cycloalkyl, - NRC(O)cycloalk 2-phenylethen-1-yl, naphthylmethyl 2-naphthylethan-1-yl, enyl, - NRC(O)substituted cycloalkenyl, - NRC(O) 2-naphthylethen-1-yl, naphthobenzyl, 2-naphthophenyle alkenyl, - NRC(O)substituted alkenyl, - NRC(O)alky than-1-yl and the like. Where specific alkyl moieties are nyl, - NRC(O)substituted alkynyl, NRC(O)aryl, intended, the nomenclature arylalkanyl, arylalkenyl and/or - NRC(O)substituted aryl, - NRC(O)heteroaryl, 40 arylalkynyl is used. In certain embodiments, an arylalkyl -NRC(O)substituted heteroaryl, - NRC(O)heterocy group is (C7-Co) arylalkyl, e.g., the alkanyl, alkenyl or alky clic, and NRC(O)substituted heterocyclic, wherein R' nyl moiety of the arylalkyl group is (C-C) and the aryl is hydrogen or alkyl and wherein alkyl, Substituted alkyl, moiety is (C-C). In certain embodiments, an arylalkyl alkenyl, Substituted alkenyl, alkynyl. Substituted alkynyl, group is (C7-Co) arylalkyl, e.g., the alkanyl, alkenyl or alky cycloalkyl, Substituted cycloalkyl, cycloalkenyl, Substituted 45 nyl moiety of the arylalkyl group is (C-Cs) and the aryl cycloalkenyl, aryl. Substituted aryl, heteroaryl, Substituted moiety is (C-C2). heteroaryl, heterocyclic, and Substituted heterocyclic are as Arylaryl by itself or as part of another substituent, refers defined herein. to a monovalent hydrocarbon group derived by the removal of “Amino” refers to the group - NH. one hydrogen atom from a single carbon atom of a ring “Substituted amino” refers to the group —NRR where 50 system in which two or more identical or non-identical aro each R is independently selected from the group consisting of matic ring systems are joined directly together by a single hydrogen, alkyl, Substituted alkyl, cycloalkyl, Substituted bond, where the number of Such direct ring junctions is one cycloalkyl, alkenyl, Substituted alkenyl, cycloalkenyl, Substi less than the number of aromatic ring systems involved. Typi tuted cycloalkenyl, alkynyl. Substituted alkynyl, aryl, het cal arylaryl groups include, but are not limited to, biphenyl, eroaryl, and heterocyclyl provided that at least one R is not 55 triphenyl, phenyl-napthyl, binaphthyl, biphenyl-napthyl, and hydrogen. the like. When the number of carbon atoms in an arylaryl “Aminoacyl refers to the group-C(O)NR'R'', wherein group is specified, the numbers refer to the carbon atoms R" and R’ independently are selected from the group con comprising each aromatic ring. For example, (C-C) ary sisting of hydrogen, alkyl, Substituted alkyl, alkenyl, Substi laryl is an arylaryl group in which each aromatic ring com tuted alkenyl, alkynyl, Substituted alkynyl, aryl, Substituted 60 prises from 5 to 14 carbons, e.g., biphenyl, triphenyl, binaph aryl, cycloalkyl, Substituted cycloalkyl, cycloalkenyl, Substi thyl, phenylnapthyl, etc. In certain embodiments, each tuted cycloalkenyl, heteroaryl, substituted heteroaryl, hetero aromatic ring system of an arylaryl group is independently a cyclic, and substituted heterocyclic and where RandR are (C-C) aromatic. In certain embodiments, each aromatic optionally joined together with the nitrogen bound thereto to ring system of an arylaryl group is independently a (Cs-Co) form a heterocyclic or substituted heterocyclic group, and 65 aromatic. In certain embodiments, each aromatic ring system wherein alkyl, substituted alkyl, alkenyl, substituted alkenyl, is identical, e.g., biphenyl, triphenyl, binaphthyl, trinaphthyl, alkynyl. Substituted alkynyl, cycloalkyl, Substituted etc. US 8,685,916 B2 11 12 “Carboxyl,” “carboxy” or “carboxylate” refers to -COH specific alkyl moieties are intended, the nomenclature het or salts thereof. eroarylalkanyl, heteroarylalkenyland/or heteroarylalkynyl is “Cyano' or "nitrile' refers to the group —CN. used. In certain embodiments, the heteroarylalkyl group is a “Cycloalkyl by itself or as part of another substituent 6-30 membered heteroarylalkyl, e.g., the alkanyl, alkenyl or refers to a saturated or unsaturated cyclic alkyl radical. Where alkynyl moiety of the heteroarylalkyl is 1-10 membered and a specific level of Saturation is intended, the nomenclature the heteroaryl moiety is a 5-20-membered heteroaryl. In cer “cycloalkanyl or “cycloalkenyl' is used. Typical cycloalkyl tain embodiments, the heteroarylalkyl group is 6-20 mem groups include, but are not limited to, groups derived from bered heteroarylalkyl, e.g., the alkanyl, alkenyl or alkynyl cyclopropane, cyclobutane, cyclopentane, cyclohexane and moiety of the heteroarylalkyl is 1-8 membered and the het the like. In certain embodiments, the cycloalkyl group is 10 eroaryl moiety is a 5-12-membered heteroaryl. (C-C) cycloalkyl. In certain embodiments, the cycloalkyl “Heterocycle.” “heterocyclic.” “heterocycloalkyl, and group is (C-C) cycloalkyl. "heterocyclyl refer to a saturated or unsaturated group hav “Cycloheteroalkyl or "heterocyclyl by itself or as part of ing a single ring or multiple condensed rings, including fused another Substituent, refers to a saturated or unsaturated cyclic bridged and spiro ring systems, and having from 3 to 15 ring alkyl radical in which one or more carbon atoms (and any 15 atoms, including 1 to 4 hetero atoms. These hetero atoms are associated hydrogenatoms) are independently replaced with selected from the group consisting of nitrogen, Sulfur, or the same or different heteroatom. Typical heteroatoms to oxygen, wherein, in fused ring systems, one or more of the replace the carbon atom(s) include, but are not limited to, N. rings can be cycloalkyl, aryl, or heteroaryl, provided that the P.O.S, Si, etc. Where a specific level of saturation is intended, point of attachment is through the non-aromatic ring. In cer the nomenclature “cycloheteroalkanyl' or “cycloheteroalk tain embodiments, the nitrogen and/or Sulfur atom(s) of the enyl' is used. Typical cycloheteroalkyl groups include, but heterocyclic group are optionally oxidized to provide for the are not limited to, groups derived from epoxides, azirines, N-oxide. —S(O)—, or —SO - moieties. thiranes, imidazolidine, morpholine, piperazine, piperidine, “Aromatic Ring System’ by itself or as part of another pyrazolidine, pyrrolidine, quinuclidine and the like. Substituent, refers to an unsaturated cyclic or polycyclic ring “Heteroalkyl, Heteroalkanyl, Heteroalkenyl and Het 25 system having a conjugated at electron system. Specifically eroalkynyl by themselves or as part of another substituent included within the definition of “aromatic ring system are refer to alkyl, alkanyl, alkenyl and alkynyl groups, respec fused ring systems in which one or more of the rings are tively, in which one or more of the carbon atoms (and any aromatic and one or more of the rings are saturated or unsat associated hydrogenatoms) are independently replaced with urated, such as, for example, fluorene, indane, indene, phe the same or different heteroatomic groups. Typical heteroat 30 nalene, etc. Typical aromatic ring systems include, but are not omic groups which can be included in these groups include, limited to, aceanthrylene, acenaphthylene, acephenan but are not limited to. —O— —S——S—S —O—S , thrylene, anthracene, azulene, benzene, chrysene, coronene, NR7R —N N , N-N , N. N. fluoranthene, fluorene, hexacene, hexaphene, hexylene, as NR'R'', PR' , P(O), , POR - O P(O), , indacene, S-indacene, indane, indene, naphthalene, octacene, S-O-, -S (O) , -SO, , –SnR'R'' and the 35 octaphene, octalene, ovalene, penta-2,4-diene, pentacene, like, where R7, R. R. R. R. R. R. and R are pentalene, pentaphene, perylene, phenalene, phenanthrene, independently hydrogen, alkyl, Substituted alkyl, aryl, Substi picene, pleiadene, pyrene, pyranthrene, rubicene, triph tuted aryl, arylalkyl, substituted arylalkyl, cycloalkyl, substi enylene, trinaphthalene and the like. tuted cycloalkyl, cycloheteroalkyl, substituted cyclohet “Heteroaromatic Ring System” by itself or as part of eroalkyl, heteroalkyl, substituted heteroalkyl, heteroaryl, 40 another Substituent, refers to an aromatic ring system in substituted heteroaryl, heteroarylalkyl or substituted het which one or more carbon atoms (and any associated hydro eroarylalkyl. gen atoms) are independently replaced with the same or dif “Heteroaryl by itself or as part of another substituent, ferent heteroatom. Typical heteroatoms to replace the carbon refers to a monovalent heteroaromatic radical derived by the atoms include, but are not limited to, N. P. O, S, Si, etc. removal of one hydrogen atom from a single atom of a het 45 Specifically included within the definition of "heteroaromatic eroaromatic ring system. Typical heteroaryl groups include, ring systems arefused ring systems in which one or more of but are not limited to, groups derived from acridine, arsindole, the rings are aromatic and one or more of the rings are satu carbazole, B-carboline, chromane, chromene, cinnoline, rated or unsaturated. Such as, for example, arsindole, benzo furan, imidazole, indazole, indole, indoline, indolizine, dioxan, benzofuran, chromane, chromene, indole, indoline, isobenzofuran, isochromene, isoindole, isoindoline, iso 50 Xanthene, etc. Typical heteroaromatic ring systems include, quinoline, isothiazole, isoxazole, naphthyridine, oxadiazole, but are not limited to, arsindole, carbazole, B-carboline, chro oxazole, perimidine, phenanthridine, phenanthroline, phena mane, chromene, cinnoline, furan, imidazole, indazole, Zine, phthalazine, pteridine, purine, pyran, pyrazine, pyra indole, indoline, indolizine, isobenzofuran, isochromene, Zole, pyridazine, pyridine, pyrimidine, pyrrole, pyrrolizine, isoindole, isoindoline, isoquinoline, isothiazole, isoxazole, quinazoline, quinoline, quinolizine, quinoxaline, tetrazole, 55 naphthyridine, oxadiazole, oxazole, perimidine, phenanthri thiadiazole, thiazole, thiophene, triazole, Xanthene, benzo dine, phenanthroline, phenazine, phthalazine, pteridine, dioxole and the like. In certain embodiments, the heteroaryl purine, pyran, pyrazine, pyrazole, pyridazine, pyridine, pyri group is from 5-20 membered heteroaryl. In certain embodi midine, pyrrole, pyrrolizine, quinazoline, quinoline, quino ments, the heteroaryl group is from 5-10 membered het lizine, quinoxaline, tetrazole, thiadiazole, thiazole, eroaryl. In certain embodiments, heteroaryl groups are those 60 thiophene, triazole, Xanthene and the like. derived from thiophene, pyrrole, benzothiophene, benzofu “Substituted” refers to a group in which one or more hydro ran, indole, pyridine, quinoline, imidazole, oxazole and pyra gen atoms are independently replaced with the same or dif Z10. ferent substituent(s). Typical substituents include, but are not “Heteroarylalkyl by itselfor as part of another substituent, limited to, alkylenedioxy (such as methylenedioxy), -M, refers to an acyclic alkyl radical in which one of the hydrogen 65 R, O, —O, OR, SR, S, S, NR'R'', atoms bonded to a carbon atom, typically a terminal or sp =NR, CF, CN, OCN, SCN, NO, NO, carbon atom, is replaced with a heteroaryl group. Where =N. —N. —S(O).O. —S(O)OH, S(O).R. OS US 8,685,916 B2 13 14 (O).O, OS(O).R., P(O)(O), P(O)(OR)(O), (i.e., a Sufficient amount of drug to effect a therapeutic effect, OP(O)(OR)(OR), C(O)R, C(S)R, C(O) e.g., a dose within the respective drugs therapeutic window, OR, C(O)NR'R'', C(O)O, C(S)OR, NRC or therapeutic range). “Multiple dose units’ or “multiples of (O)NR'R'', NRC(S)NR'R'', NRPC(NR) a dose unit' or a “multiple of a dose unit refers to at least two NR'R'' and C(NR)NR'R' where M is halogen; R, single dose units. R', R'' and Rare independently hydrogen, alkyl, substi “Gastrointestinal enzyme” or “GI enzyme” refers to an tuted alkyl, alkoxy, Substituted alkoxy, cycloalkyl, Substituted enzyme located in the gastrointestinal (GI) tract, which cycloalkyl, cycloheteroalkyl, substituted cycloheteroalkyl, encompasses the anatomical sites from mouth to anus. aryl, substituted aryl, heteroaryl or substituted heteroaryl, or Trypsin is an example of a GI enzyme. optionally R'' and R' together with the nitrogen atom to 10 “Gastrointestinal enzyme-cleavable moiety' or “GI which they are bonded form a cycloheteroalkyl or substituted enzyme-cleavable moiety refers to a group comprising a site cycloheteroalkyl ring; and R and R are independently Susceptible to cleavage by a GI enzyme. For example, a hydrogen, alkyl, Substituted alkyl, aryl, cycloalkyl, Substi "trypsin-cleavable moiety refers to a group comprising a site tuted cycloalkyl, cycloheteroalkyl, substituted cyclohet Susceptible to cleavage by trypsin. eroalkyl, aryl, substituted aryl, heteroaryl or substituted het 15 “Gastrointestinal enzyme inhibitor or “GI enzyme inhibi eroaryl, or optionally RandR together with the nitrogen tor” refers to any agent capable of inhibiting the action of a atom to which they are bonded form a cycloheteroalkyl or gastrointestinal enzyme on a substrate. The term also encom Substituted cycloheteroalkyl ring. In certain embodiments, passes salts of gastrointestinal enzyme inhibitors. For substituents include -M, R', example, a "trypsin inhibitor refers to any agent capable of —O, OR, SR, S, —S, NR'R'', —NR, inhibiting the action of trypsin on a substrate. CF, CN, OCN, SCN, NO, NO, —N, “Patient includes humans, and also other mammals, such N, S(O) R', OS(O)O, OS(O).R., P(O) as livestock, Zoo animals, and companion animals. Such as a (O), P(O)(OR)(O), OP(O)(OR)(OR), cat, dog, or horse. C(O)R, C(S)R, C(O)OR, C(O)NR'R'', "Pharmaceutical composition” refers to at least one com C(O)O, NRC(O)NR'R''. In certain embodi 25 pound and can further comprise a pharmaceutically accept ments, substituents include -M, R', able carrier, with which the compound is administered to a —O, OR, SR, NR'R'', CF, CN, NO, patient. S(O).R, P(O)(OR)(O), OP(O)(OR)(OR), "Pharmaceutically acceptable carrier refers to a diluent, C(O)R, C(O)OR, C(O)NR'R'', C(O)O. adjuvant, excipient or vehicle with, or in which a compound In certain embodiments, substituents include -M, R', 30 is administered. —O, OR, SR, NR'R'', CF, CN, NO, "Pharmaceutically acceptable salt” refers to a salt of a S(O).R, OP(O)(OR)(OR), C(O)R', C(O) compound, which possesses the desired pharmacological OR, C(O)O, where R, R and Rare as defined activity of the compound. Such salts include: (1) acid addition above. For example, a Substituted group may bear a meth salts, formed with inorganic acids such as hydrochloric acid, ylenedioxy substituent or one, two, or three substituents 35 hydrobromic acid, Sulfuric acid, nitric acid, phosphoric acid, Selected from a halogen atom, a (1-4C)alkyl group and a and the like; or formed with organic acids such as acetic acid, (1-4C)alkoxy group. propionic acid, hexanoic acid, cyclopentanepropionic acid, It is understood that in all substituted groups defined above, glycolic acid, pyruvic acid, lactic acid, malonic acid, Succinic polymers arrived at by defining substituents with further sub acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric stituents to themselves (e.g., Substituted aryl having a Substi 40 acid, benzoic acid, 3-(4-hydroxybenzoyl)benzoic acid, cin tuted aryl group as a substituent which is itself substituted namic acid, mandelic acid, methanesulfonic acid, ethane with a substituted aryl group, which is further substituted by Sulfonic acid, 1.2-ethane-disulfonic acid, 2-hydroxyethane a Substituted aryl group, etc.) are not intended for inclusion sulfonic acid, benzenesulfonic acid, herein. In Such cases, the maximum number of Such substi 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, tutions is three. For example, serial substitutions of substi 45 4-toluenesulfonic acid, camphorsulfonic acid, 4-methylbicy tuted aryl groups are limited to substituted aryl-(substituted clo2.2.2-oct-2-ene-1-carboxylic acid, glucoheptonic acid, aryl)-substituted aryl. 3-phenylpropionic acid, trimethylacetic acid, tertiary buty As to any of the groups disclosed herein which contain one lacetic acid, lauryl Sulfuric acid, gluconic acid, glutamic acid, or more Substituents, it is understood, of course, that Such hydroxynaphthoic acid, Salicylic acid, Stearic acid, muconic groups do not contain any Substitution or Substitution patterns 50 acid, and the like; or (2) salts formed when an acidic proton which are sterically impractical and/or synthetically non present in the compound is replaced by a metal ion, e.g., an feasible. In addition, the Subject compounds include all Ste alkali metalion, an alkaline earth ion, oran aluminum ion; or reochemical isomers arising from the Substitution of these coordinates with an organic base Such as ethanolamine, compounds. diethanolamine, triethanolamine, N-methylglucamine and Unless indicated otherwise, the nomenclature of substitu 55 the like. ents that are not explicitly defined herein are arrived at by "Pharmacodynamic (PD) profile' refers to a profile of the naming the terminal portion of the functionality followed by efficacy of a drug in a patient (or Subject or user), which is the adjacent functionality toward the point of attachment. For characterized by PD parameters. “PD parameters' include example, the substituent “arylalkyloxycarbonyl refers to the “drug Emax” (the maximum drug efficacy), “drug EC50 (the group (aryl)-(alkyl)-O-C(O)—. 60 concentration of drug at 50% of the Emax) and side effects. “Dose unit as used herein refers to a combination of a GI “PK parameter refers to a measure of drug concentration enzyme-cleavable prodrug (e.g., trypsin-cleavable prodrug) in blood or plasma, such as: 1) "drug Cmax', the maximum and a GI enzyme inhibitor (e.g., a trypsin inhibitor). A 'single concentration of drug achieved in blood or plasma; 2) "drug dose unit is a single unit of a combination of a GI enzyme Tmax', the time elapsed following ingestion to achieve cleavable prodrug (e.g., trypsin-cleavable prodrug) and a GI 65 Cmax; and 3)"drug exposure', the total concentration of drug enzyme inhibitor (e.g., trypsin inhibitor), where the single present in blood or plasma over a selected period of time, dose unit provide a therapeutically effective amount of drug which can be measured using the area under the curve (AUC) US 8,685,916 B2 15 16 of a time course of drug release over a selected period of time “solely.” “only' and the like in connection with the recitation (t). Modification of one or more PK parameters provides for of claim elements, or use of a “negative' limitation. a modified PK profile. It should be understood that as used herein, the term “a” “PK profile” refers to a profile of drug concentration in entity or “an entity refers to one or more of that entity. For blood or plasma. Such a profile can be a relationship of drug 5 example, a compound refers to one or more compounds. As concentration over time (i.e., a “concentration-time PK pro such, the terms “a”, “an”, “one or more' and “at least one' can file') or a relationship of drug concentration versus number of be used interchangeably. Similarly the terms “comprising, doses ingested (i.e., a “concentration-dose PK profile'). APK “including and “having can be used interchangeably. profile is characterized by PK parameters. The publications discussed herein are provided solely for “Preventing or “prevention” or “prophylaxis' refers to a 10 their disclosure prior to the filing date of the present applica reduction in risk of occurrence of a condition, such as pain. “Prodrug” refers to a derivative of an active agent that tion. Nothing herein is to be construed as an admission that requires a transformation within the body to release the active the present invention is not entitled to antedate Such publica agent. In certain embodiments, the transformation is an enzy tion by virtue of prior invention. Further, the dates of publi matic transformation. In certain embodiments, the transfor 15 cation provided may be different from the actual publication mation is a cyclization transformation. In certain embodi dates which may need to be independently confirmed. ments, the transformation is a combination of an enzymatic Unless defined otherwise, all technical and scientific terms transformation and a cyclization transformation. Prodrugs used herein have the same meaning as commonly understood are frequently, although not necessarily, pharmacologically by one of ordinary skill in the art to which this invention inactive until converted to the active agent. belongs. Although any methods and materials similar or “Promoiety” refers to a form of protecting group that when equivalent to those described herein can also be used in the used to mask a functional group within an active agent con practice or testing of the present invention, the preferred verts the active agent into a prodrug. Typically, the promoiety methods and materials are now described. All publications will be attached to the drug via bond(s) that are cleaved by mentioned herein are incorporated herein by reference to enzymatic or non-enzymatic means in vivo. 25 disclose and describe the methods and/or materials in con “Solvate” as used herein refers to a complex or aggregate nection with which the publications are cited. formed by one or more molecules of a solute, e.g. a prodrug or Except as otherwise noted, the methods and techniques of a pharmaceutically acceptable salt thereof, and one or more the present embodiments are generally performed according molecules of a solvent. Such solvates are typically crystalline to conventional methods well known in the art and as Solids having a Substantially fixed molar ratio of Solute and 30 described in various general and more specific references that solvent. Representative solvents include by way of example, are cited and discussed throughout the present specification. water, methanol, ethanol, isopropanol, acetic acid, and the See, e.g., Loudon, Organic Chemistry, Fourth Edition, New like. When the solvent is water, the solvate formed is a York: Oxford University Press, 2002, pp. 360-361, 1084 hydrate. 1085; Smith and March, March's Advanced Organic Chem “Therapeutically effective amount’ means the amount of a 35 istry: Reactions, Mechanisms, and Structure, Fifth Edition, compound (e.g., prodrug) that, when administered to a patient Wiley-Interscience, 2001. for preventing or treating a condition Such as pain, is suffi The nomenclature used herein to name the Subject com cient to effect such treatment. The “therapeutically effective pounds is illustrated in the Examples herein. In certain amount will vary depending on the compound, the condition instances, this nomenclature has is derived using the commer and its severity and the age, weight, etc., of the patient. 40 cially-available AutoNom software (MDL, San Leandro, “Treating or “treatment of any condition, such as pain, Calif.). refers, in certain embodiments, to ameliorating the condition It is appreciated that certain features of the invention, (i.e., arresting or reducing the development of the condition). which are, for clarity, described in the context of separate In certain embodiments “treating” or “treatment” refers to embodiments, may also be provided in combination in a ameliorating at least one physical parameter, which may not 45 single embodiment. Conversely, various features of the be discernible by the patient. In certain embodiments, “treat invention, which are, for brevity, described in the context of a ing or “treatment” refers to inhibiting the condition, either single embodiment, may also be provided separately or in any physically, (e.g., stabilization of a discernible symptom), suitable sub-combination. All combinations of the embodi physiologically, (e.g., stabilization of a physical parameter), ments pertaining to the chemical groups represented by the or both. In certain embodiments, “treating or “treatment' 50 variables are specifically embraced by the present invention refers to delaying the onset of the condition. and are disclosed hereinjust as if each and every combination was individually and explicitly disclosed, to the extent that DETAILED DESCRIPTION Such combinations embrace compounds that are stable com pounds (i.e., compounds that can be isolated, characterised, Before the present invention is further described, it is to be 55 and tested for biological activity). In addition, all Sub-com understood that this invention is not limited to particular binations of the chemical groups listed in the embodiments embodiments described, as Such may, of course, vary. It is describing Such variables are also specifically embraced by also to be understood that the terminology used herein is for the present invention and are disclosed herein just as if each the purpose of describing particular embodiments only, and is and every Such sub-combination of chemical groups was not intended to be limiting, since the scope of the present 60 individually and explicitly disclosed herein. invention will be limited only by the appended claims. General Synthetic Procedures It must be noted that as used herein and in the appended Many general references providing commonly known claims, the singular forms “a,” “an,” and “the include plural chemical synthetic schemes and conditions useful for synthe referents unless the context clearly dictates otherwise. It is sizing the disclosed compounds are available (see, e.g., Smith further noted that the claims may be drafted to exclude any 65 and March, March's Advanced Organic Chemistry: Reac optional element. As such, this statement is intended to serve tions, Mechanisms, and Structure, Fifth Edition, Wiley-Inter as antecedent basis for use of Such exclusive terminology as science, 2001; or Vogel, A Textbook of Practical Organic US 8,685,916 B2 17 18 Chemistry, Including Qualitative Organic Analysis, Fourth configuration of the spacer leaving group being Such that, Edition, New York: Longman, 1978). upon enzymatic cleavage of the cleavable moiety, the nucleo Compounds as described herein can be purified by any of philic nitrogen is capable of forming a cyclic urea, liberating the means known in the art, including chromatographic the compound from the spacer leaving group so as to provide means. Such as high performance liquid chromatography the patient with controlled release of an opioid. (HPLC), preparative thin layer chromatography, flash col The corresponding compound (prodrug inaccordance with umn chromatography and ion exchange chromatography. the present disclosure) provides post administration-acti Any suitable stationary phase can be used, including normal vated, controlled release of an opioid, because it requires and reversed phases as well as ionic resins. See, e.g., Intro 10 enzymatic cleavage to initiate release of the compound, and duction to Modern Liquid Chromatography, 2nd Edition, ed. because the rate of release of the opioid depends upon both L. R. Snyder and J. J. Kirkland, John Wiley and Sons, 1979; the rate of enzymatic cleavage and the rate of cyclization. The and Thin Layer Chromatography, ed. E. Stahl, Springer-Ver prodrug is configured so that it will not provide excessively lag, New York, 1969. high plasma levels of the opioid if it is administered inappro During any of the processes for preparation of the com 15 priately, and cannot readily be decomposed to afford the pounds of the present disclosure, it may be necessary and/or opioid other than by enzymatic cleavage followed by con trolled cyclization. desirable to protect sensitive or reactive groups on any of the The enzyme capable of cleaving the enzyme-cleavable molecules concerned. This can be achieved by means of moiety can be a peptidase, also referred to as a protease—the conventional protecting groups as described in standard enzyme-cleavable moiety being linked to the nucleophilic works, such as T. W. Greene and P. G. M. Wuts, “Protective nitrogen through an amide (e.g. a peptide: —NHCO—) bond. Groups in Organic Synthesis'. Fourth edition, Wiley, New In some embodiments, the enzyme is a digestive enzyme, York 2006. The protecting groups can be removed at a con Such as a digestive enzyme of a protein. Venient Subsequent stage using methods known from the art. The enzyme-cleavable moiety linked to the nucleophilic The compounds described herein can contain one or more 25 nitrogen through an amide bond can be, for example, a resi chiral centers and/or double bonds and therefore, can exist as due of an amino acid or a peptide, a variant of a residue of an Stereoisomers, such as double-bond isomers (i.e., geometric amino acid or a peptide, a derivative of a residue of an amino isomers), enantiomers or diastereomers. Accordingly, all pos acid or a peptide, or a derivative of a residue of an amino acid sible enantiomers and stereoisomers of the compounds 30 variant or a peptide variant. As discussed below, an amino including the stereoisomerically pure form (e.g., geometri acid variant refers to an amino acid other than any of the 20 cally pure, enantiomerically pure or diastereomerically pure) common naturally occurring L-amino acids that is hydrolyz and enantiomeric and stereoisomeric mixtures are included in able by a protease in a manner similar to the ability of a the description of the compounds herein. Enantiomeric and protease to hydrolyze a naturally occurring L-amino acid. A Stereoisomeric mixtures can be resolved into their component 35 derivative refers to a substance that has been altered from enantiomers or stereoisomers using separation techniques or another Substance by modification, partial Substitution, chiral synthesis techniques well known to the skilled artisan. homologation, truncation, or a change in oxidation state. For The compounds can also exist in several tautomeric forms example, an N-acyl derivative of an amino acid is an example including the enol form, the keto form and mixtures thereof. of a derivative of an amino acid. Accordingly, the chemical structures depicted herein encom 40 In some instances, the enzyme-cleavable moiety can be an pass all possible tautomeric forms of the illustrated com (alpha) N-acyl derivative of an amino acid or peptide or an pounds. (alpha) N-acyl derivative of an amino acid variant or peptide The compounds described also include isotopically variant. labeled compounds where one or more atoms have an atomic The peptide can contain, for example, up to about 100 mass different from the atomic mass conventionally found in 45 amino acid residues. Each amino acid can advantageously be nature. Examples of isotopes that can be incorporated into the a naturally occurring amino acid, such as an L-amino acid. compounds disclosed herein include, but are not limited to, Examples of naturally occurring amino acids are alanine, 2H, H, C, 13C, C, 15N, 18O, 17O, etc. Compounds can arginine, asparagine, aspartic acid, cysteine, glutamic acid, exist in unsolvated forms as well as Solvated forms, including glutamine, glycine, histidine, isoleucine, leucine, lysine, hydrated forms. In general, compounds can be hydrated or 50 methionine, phenylalanine, proline, serine, threonine, tryp Solvated. Certain compounds can exist in multiple crystalline tophan, tyrosine and valine. Accordingly, examples of or amorphous forms. In general, all physical forms are enzyme-cleavable moieties include residues of the L-amino equivalent for the uses contemplated herein and are intended acids listed herein and N-acyl derivatives thereof, and pep to be within the scope of the present disclosure. tides formed from at least two of the L-amino acids listed 55 herein and the N-acyl derivatives thereof. Additional Representative Embodiments examples include residues of amino acid variants and N-acyl Reference will now be made in detail to various embodi derivatives thereof, and peptides formed from at least two of ments. It will be understood that the invention is not limited to the L-amino acids listed above and/or variants thereof, and these embodiments. To the contrary, it is intended to cover N-acyl derivatives thereof. Also included are derivatives of alternatives, modifications, and equivalents as may be 60 Such amino acids oramino acid variants and peptides thereof. included within the spirit and scope of the allowed claims. The embodiments provide a prodrug with a substituent The disclosure provides a method of providing a patient which is a spacer leaving group bearing a nucleophilic nitro with post administration-activated, controlled release of an gen that is protected with an enzyme-cleavable moiety. Upon opioid, which comprises administering to the patient a corre enzymatic cleavage of the cleavable moiety, the nucleophilic sponding compound in which the opioid has a Substituent 65 nitrogen is capable of forming a cyclic urea. A representative which is a spacer leaving group bearing a nucleophilic nitro scheme of a cyclization of a spacer group is shown below, gen that is protected with an enzyme-cleavable moiety, the wherein X is an opioid. US 8,685,916 B2 19 20 The present embodiments provide a compound of formula I:

(I) l R R2 O R3 R6 R7 (Y), X N an N --- N N 10 H R5 O R3 (Y),

15 wherein (Y), X is selected from a residue of a ketone-containing opioid, wherein the hydrogen atom of the corresponding hydroxyl group of the enolic tautomer of the ketone is replaced by a The rate of cyclization of the cyclic urea can be adjusted by covalent bond to C(O) NI (A ring)-Y. (CRR), incorporation of a heterocyclic ring within the spacer group. NH CO) CH(R) N(R) C(O) CH(R) In certain embodiments, incorporation of a heterocyclic ring N(R) R'; a residue of a phenolic opioid, wherein the within the spacer group results in formation of a fused ring hydrogenatom of the phenolic hydroxyl group is replaced by cyclic urea and in a faster cyclization reaction. a covalent bond to —C(O) N(A ring)-Y. (CRR) NH CO) CH(R) N(R) C(O) CH(R) The cyclic group formed when the opioid is released is 25 N(R) R', and a residue of an amide-containing opioid, conveniently pharmaceutically acceptable, in particular a wherein - C(O) N(A ring)-Y-(CRR). NH C pharmaceutically acceptable cyclic urea. It will be appreci (O) CH(R) N(R) C(O) CH(R) N(R) R7 is ated that cyclic ureas are generally very stable and have low connected to the amide-containing opioid through the oxygen toxicity. of the amide group, wherein the amide group is converted to 30 an amide enol or an imine tautomer, According to one aspect, the embodiments include phar the A ring is a heterocyclic 5 to 12-membered ring: maceutical compositions, which comprise a GI enzyme each Y is independently selected from alkyl, substituted cleavable opioid prodrug and an optional GI enzyme inhibi alkyl, alkenyl, Substituted alkenyl, alkynyl, Substituted alky tor. Examples of opioid prodrugs and enzyme inhibitors are nyl, aryl, Substituted aryl, acyl, Substituted acyl, carboxyl, described below. 35 alkoxycarbonyl, Substituted alkoxycarbonyl, aminoacyl, Sub stituted aminoacyl, amino, Substituted amino, acylamino, Opioid Prodrugs Substituted acylamino, and cyano; An “opioid refers to a chemical substance that exerts its c is a number from Zero to 3: pharmacological action by interaction at an opioid receptor. each R' is independently selected from hydrogen, alkyl, 40 substituted alkyl, alkenyl, substituted alkenyl, alkynyl, sub An opioid can be a natural product, a synthetic compound or stituted alkynyl, aryl, Substituted aryl, acyl, Substituted acyl, a semi-synthetic compound. In certain embodiments, an carboxyl, alkoxycarbonyl, Substituted alkoxycarbonyl, ami opioid is a compound with a pharmacophore that presents to noacyl, Substituted aminoacyl, amino, Substituted amino, the opioid receptor an aromatic group and an aliphatic amine acylamino, Substituted acylamino, and cyano; group in an architecturally discrete way. See, for example, 45 each R is independently selected from hydrogen, alkyl, Foye's Principles of Medicinal Chemistry, Sixth Edition, ed. substituted alkyl, alkenyl, substituted alkenyl, alkynyl, sub stituted alkynyl, aryl, Substituted aryl, acyl, Substituted acyl, T. L. Lemke and D. A. Williams, Lippincott Williams & carboxyl, alkoxycarbonyl, Substituted alkoxycarbonyl, ami Wilkins, 2008, particularly Chapter 24, pages 653-678. noacyl, Substituted aminoacyl, amino, Substituted amino, The disclosure provides an opioid prodrug that provides 50 acylamino, Substituted acylamino, and cyano; or controlled release of an opioid. The disclosure provides a R" and R together with the carbon to which they are attached can form a cycloalkyl or Substituted cycloalkyl promoiety that is attached to an opioid through any structural group, or two R' or R groups on adjacent carbon atoms, moiety on the opioid, where the structural moiety has a reac together with the carbonatoms to which they are attached, can tive group. Examples of reactive groups on an opioid include, 55 form a cycloalkyl or Substituted cycloalkyl group; but are not limited to ketone, phenol, and amide. a is an integer from one to 8: It is contemplated that opioids bearing at least some of the provided that whena is one, the A ring is a heterocyclic 6 to functionalities described herein will be developed; such opio 12-membered ring; and when the A ring is a heterocyclic 5-membered ring, then a is an integer from 2 to 8; ids are included as part of the scope of this disclosure. 60 each R is independently hydrogen, alkyl, substituted Formula I alkyl, aryl or substituted aryl; R is selected from hydrogen, alkyl, substituted alkyl, aryl, Compounds of the present disclosure include compounds substituted aryl, arylalkyl, substituted arylalkyl, heteroalkyl, of formula I shown below. Compositions of the present dis substituted heteroalkyl, heteroaryl, substituted heteroaryl, closure also include compounds of formula I shown below. 65 heteroarylalkyl, and substituted heteroarylalkyl: Pharmaceutical compositions and methods of the present dis each R is independently selected from hydrogen, alkyl, closure also contemplate compounds of formula I. substituted alkyl, aryl, substituted aryl, arylalkyl, substituted US 8,685,916 B2 21 22 arylalkyl, heteroalkyl, substituted heteroalkyl, heteroaryl, As disclosed herein, an enzyme-cleavable ketone-modified substituted heteroaryl, heteroarylalkyl, and substituted het opioid prodrug is a ketone-modified opioid prodrug that com eroarylalkyl: prises a promoiety comprising an enzyme-cleavable moiety, b is a number from Zero to 100; and i.e., a moiety having a site Susceptible to cleavage by an R’ is selected from hydrogen, alkyl, substituted alkyl, acyl, enzyme. In one embodiment, the cleavable moiety is a GI Substituted acyl, alkoxycarbonyl, Substituted alkoxycarbo enzyme-cleavable moiety, such as a trypsin-cleavable moiety. nyl, aryl, Substituted aryl, arylalkyl, and Substituted arylalkyl; Such a prodrug comprises a ketone-containing opioid or a salt, hydrate or solvate thereof. covalently bound to a promoiety comprising an enzyme In formula I, X can be selected from a residue of a ketone cleavable moiety, wherein cleavage of the enzyme-cleavable containing opioid, wherein the hydrogen atom of the corre 10 moiety by an enzyme mediates release of the drug. sponding hydroxyl group of the enolic tautomer of the ketone Formulae II-V is replaced by a covalent bond to —C(O)—NI (A ring)-Y— Compounds of the present disclosure include compounds (CRR). NH CO) CH(R) N(R) C(O) CH of formulae II-V shown below. Compositions of the present (R) N(R) R': a residue of a phenolic opioid, wherein disclosure also include compounds of formulae II-V shown the hydrogenatom of the phenolic hydroxyl group is replaced 15 below. Pharmaceutical compositions and methods of the by a covalent bond to C(O) NLA ring)-Y. (CRR), present disclosure also contemplate compounds of formulae NH C(O) CH(R) N(R) C(O) CH(R) II-V. N(R) R', and a residue of an amide-containing opioid, The present embodiments provide a compound of formula wherein –C(O) N(A ring)-Y-(CRR). NH C II: (O) CH(R) N(R)-C(O)-CH(R) N(R) R7 is connected to the amide-containing opioid through the oxygen of the amide group, wherein the amide group is converted to (II) an amide enol or an imine tautomer. l R1 R2 O R3 R6 In certain instances, X is a ketone-containing opioid, 25 R7 wherein the hydrogen atom of the corresponding hydroxyl X N d N N group of the enolic tautomer of the ketone is replaced by a A covalent bond to —C(O) NLA ring)-Y. (CRR) H R5 O R3 NH C(O) CH(R) N(R) C(O) CH(R) (Y), N(R)) R7. 30 In certain instances, X is a ketone-containing opioid, wherein the opioid is selected from acetylmorphone, hydro wherein codone, hydromorphone, ketobemidone, methadone, nalox X represents a residue of a ketone-containing opioid, one, naltrexone, N-methylnaloxone, N-methylmaltrexone, wherein the hydrogen atom of the corresponding hydroxyl oxycodone, oxymorphone, and pentamorphone. 35 group of the enolic tautomer of the ketone is replaced by a In certain instances, X is a residue of a phenolic opioid, covalent bond to C(O) NI (A ring)-Y. (CRR), wherein the hydrogenatom of the phenolic hydroxyl group is NH CO) CH(R) N(R) C(O) CH(R) replaced by a covalent bond to —C(O)—N(A ring)-Y— N(R) R7: (CRR). NH CO) CH(R) N(R) C(O) CH the A ring is a heterocyclic 5 to 12-membered ring: (R) N(R) R7. 40 each Y is independently selected from alkyl, substituted In certain instances, X is a phenolic opioid, wherein the alkyl, alkenyl, Substituted alkenyl, alkynyl, Substituted alky opioid is selected from buprenorphine, dihydroetorphine, nyl, aryl, Substituted aryl, acyl, Substituted acyl, carboxyl, diprenorphine, etorphine, hydromorphone, levorphanol, alkoxycarbonyl, Substituted alkoxycarbonyl, aminoacyl, Sub morphine, nalbuphine, nalmefene, nalorphine, naloxone, nal stituted aminoacyl, amino, Substituted amino, acylamino, trexone, N-methyldiprenorphine, N-methylnaloxone, N-me 45 Substituted acylamino, and cyano; thylmaltrexone, oripavine, oxymorphone, butorphanol, dezo c is a number from Zero to 3: cine, ketobemidone, meptazinol, o-desmethyltramadol, each R' is independently selected from hydrogen, alkyl, pentazocine, phenazocine, and tapentadol. substituted alkyl, alkenyl, substituted alkenyl, alkynyl, sub In certain instances, X is a residue of an amide-containing stituted alkynyl, aryl, Substituted aryl, acyl, Substituted acyl, opioid, wherein - C(O) N (A ring)-Y. (CRR), 50 carboxyl, alkoxycarbonyl, Substituted alkoxycarbonyl, ami NH C(O) CH(R) N(R) C(O) CH(R) noacyl, Substituted aminoacyl, amino, Substituted amino, N(R) R' is connected to the amide-containing opioid acylamino, Substituted acylamino, and cyano; through the oxygen of the amide group, wherein the amide each R is independently selected from hydrogen, alkyl, group is converted to an amide enol or an imine tautomer. substituted alkyl, alkenyl, substituted alkenyl, alkynyl, sub In certain instances, X is an amide-containing opioid, 55 stituted alkynyl, aryl, Substituted aryl, acyl, Substituted acyl, wherein the opioid is selected from alfentanil, carfentanil, carboxyl, alkoxycarbonyl, Substituted alkoxycarbonyl, ami fentanyl, lofentanil, loperamide, olmefentanyl, remifentanil, noacyl, Substituted aminoacyl, amino, Substituted amino, and Sufentanil. acylamino, Substituted acylamino, and cyano; or Ketone-modified Opioid Prodrugs R" and R together with the carbon to which they are The disclosure provides a ketone-modified opioid prodrug 60 attached can form a cycloalkyl or Substituted cycloalkyl that provides controlled release of a ketone-containing group, or two R' or R groups on adjacent carbon atoms, opioid. In a ketone-modified opioid prodrug, a promoiety is together with the carbonatoms to which they are attached, can attached to the ketone-containing opioid through the enolic form a cycloalkyl or Substituted cycloalkyl group; oxygen atom of the ketone moiety. In a ketone-modified a is an integer from one to 8: opioid prodrug, the hydrogen atom of the corresponding 65 provided that whena is one, the A ring is a heterocyclic 6 to hydroxyl group of the enolic tautomer of the ketone-contain 12-membered ring; and when the A ring is a heterocyclic ing opioid is replaced by a covalent bond to a promoiety. 5-membered ring, then a is an integer from 2 to 8; US 8,685,916 B2 23 24 each R is independently hydrogen, alkyl, substituted a is an integer from one to 8: alkyl, aryl or substituted aryl; provided that whena is one, the A ring is a heterocyclic 6 to R is selected from hydrogen, alkyl, substituted alkyl, aryl, 12-membered ring; and when the A ring is a heterocyclic substituted aryl, arylalkyl, substituted arylalkyl, heteroalkyl, substituted heteroalkyl, heteroaryl, substituted heteroaryl, 5-membered ring, then a is an integer from 2 to 8; heteroarylalkyl, and substituted heteroarylalkyl; each R is independently hydrogen, alkyl, substituted each R is independently selected from hydrogen, alkyl, alkyl, aryl or substituted aryl; substituted alkyl, aryl, substituted aryl, arylalkyl, substituted R is selected from hydrogen, alkyl, substituted alkyl, aryl, arylalkyl, heteroalkyl, substituted heteroalkyl, heteroaryl, 10 substituted aryl, arylalkyl, substituted arylalkyl, heteroalkyl, substituted heteroaryl, heteroarylalkyl, and substituted het substituted heteroalkyl, heteroaryl, substituted heteroaryl, eroarylalkyl: heteroarylalkyl, and substituted heteroarylalkyl: b is a number from Zero to 100; and R’ is selected from hydrogen, alkyl, substituted alkyl, acyl, each R is independently selected from hydrogen, alkyl, Substituted acyl, alkoxycarbonyl, Substituted alkoxycarbo 15 substituted alkyl, aryl, substituted aryl, arylalkyl, substituted nyl, aryl, Substituted aryl, arylalkyl, and Substituted arylalkyl; arylalkyl, heteroalkyl, substituted heteroalkyl, heteroaryl, or a salt, hydrate or solvate thereof. substituted heteroaryl, heteroarylalkyl, and substituted het The present embodiments provide a compound of formula eroarylalkyl: III: b is a number from Zero to 100; and

(III)

(Y),

wherein R’ is selected from hydrogen, alkyl, substituted alkyl, acyl, R" is hydrogen or hydroxyl; Substituted acyl, alkoxycarbonyl, Substituted alkoxycarbo R’ is hydrogen or alkyl; nyl, aryl, Substituted aryl, arylalkyl, and Substituted arylalkyl; the A ring is a heterocyclic 5 to 12-membered ring; 40 or a salt, hydrate or solvate thereof. each Y is independently selected from alkyl, substituted The present embodiments provide a compound of formula alkyl, alkenyl, Substituted alkenyl, alkynyl, Substituted alky IV: nyl, aryl, Substituted aryl, acyl, Substituted acyl, carboxyl, alkoxycarbonyl. Substituted alkoxycarbonyl, aminoacyl, Sub 45 stituted aminoacyl, amino, Substituted amino, acylamino, Substituted acylamino, and cyano; l R1 R2 O R3 R6 c is a number from Zero to 3: R7 X N d N each R" is independently selected from hydrogen, alkyl, N substituted alkyl, alkenyl, substituted alkenyl, alkynyl, sub 50 A stituted alkynyl, aryl, Substituted aryl, acyl, Substituted acyl, H R5 O R3 carboxyl, alkoxycarbonyl, Substituted alkoxycarbonyl, ami (Y), noacyl, Substituted aminoacyl, amino, Substituted amino, acylamino, Substituted acylamino, and cyano; 55 wherein each R is independently selected from hydrogen, alkyl, X represents a residue of a ketone-containing opioid, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, sub wherein the hydrogen atom of the corresponding hydroxyl stituted alkynyl, aryl, Substituted aryl, acyl, Substituted acyl, group of the enolic tautomer of the ketone is replaced by a carboxyl, alkoxycarbonyl, Substituted alkoxycarbonyl, ami covalent bond to C(O) NI (A ring)-Y. (CRR), noacyl, Substituted aminoacyl, amino, Substituted amino, 60 NH CO) CH(R) N(R) C(O) CH(R) acylamino, Substituted acylamino, and cyano; or N(R) R7: R" and R together with the carbon to which they are the A ring is a heterocyclic 5 to 12-membered ring: attached can form a cycloalkyl or Substituted cycloalkyl each Y is independently selected from alkyl, substituted group, or two R' or R groups on adjacent carbon atoms, 65 alkyl, alkenyl, Substituted alkenyl, alkynyl, Substituted alky together with the carbonatoms to which they are attached, can nyl, aryl, Substituted aryl, acyl, Substituted acyl, carboxyl, form a cycloalkyl or Substituted cycloalkyl group; alkoxycarbonyl, Substituted alkoxycarbonyl, aminoacyl, Sub US 8,685,916 B2 25 26 stituted aminoacyl, amino, Substituted amino, acylamino, covalent bond to C(O) NI (A ring)-Y. (CRR), Substituted acylamino, and cyano; NH CO) CH(R) N(R) C(O) CH(R) c is a number from Zero to 3: N(R) R7: each R" is independently selected from hydrogen, alkyl, the A ring is a heterocyclic 5 to 12-membered ring: substituted alkyl, alkenyl, substituted alkenyl, alkynyl, sub each Y is independently selected from alkyl, substituted stituted alkynyl, aryl, Substituted aryl, acyl, Substituted acyl, alkyl, alkenyl, Substituted alkenyl, alkynyl, Substituted alky carboxyl, alkoxycarbonyl, Substituted alkoxycarbonyl, ami nyl, aryl, Substituted aryl, acyl, Substituted acyl, carboxyl, noacyl, Substituted aminoacyl, amino, Substituted amino, alkoxycarbonyl, Substituted alkoxycarbonyl, aminoacyl, Sub acylamino, Substituted acylamino, and cyano; 10 stituted aminoacyl, amino, Substituted amino, acylamino, each R is independently selected from hydrogen, alkyl, Substituted acylamino, and cyano; substituted alkyl, alkenyl, substituted alkenyl, alkynyl, sub c is a number from Zero to 3: stituted alkynyl, aryl, Substituted aryl, acyl, Substituted acyl, each R' is independently selected from hydrogen, alkyl, carboxyl, alkoxycarbonyl, Substituted alkoxycarbonyl, ami substituted alkyl, alkenyl, substituted alkenyl, alkynyl, sub noacyl, Substituted aminoacyl, amino, Substituted amino, 15 stituted alkynyl, aryl, Substituted aryl, acyl, Substituted acyl, acylamino, Substituted acylamino, and cyano; or carboxyl, alkoxycarbonyl, Substituted alkoxycarbonyl, ami R" and R together with the carbon to which they are noacyl, Substituted aminoacyl, amino, Substituted amino, attached can form a cycloalkyl or Substituted cycloalkyl acylamino, Substituted acylamino, and cyano; group, or two R' or R groups on adjacent carbon atoms, each R is independently selected from hydrogen, alkyl, together with the carbonatoms to which they are attached, can substituted alkyl, alkenyl, substituted alkenyl, alkynyl, sub form a cycloalkyl or Substituted cycloalkyl group; stituted alkynyl, aryl, Substituted aryl, acyl, Substituted acyl, a is an integer from one to 8: carboxyl, alkoxycarbonyl, Substituted alkoxycarbonyl, ami provided that whena is one, the Aring is a heterocyclic 6 to noacyl, Substituted aminoacyl, amino, Substituted amino, 12-membered ring; and when the A ring is a heterocyclic 25 acylamino, Substituted acylamino, and cyano; or 5-membered ring, then a is an integer from 2 to 8: R" and R together with the carbon to which they are attached can form a cycloalkyl or Substituted cycloalkyl each R is independently hydrogen, alkyl, substituted group, or two R' or R groups on adjacent carbon atoms, alkyl, aryl or substituted aryl; together with the carbonatoms to which they are attached, can R is a side chain of an amino acid selected from alanine, 30 form a cycloalkyl or Substituted cycloalkyl group; arginine, asparagine, aspartic acid, cysteine, glutamic acid, a is an integer from one to 8: glutamine, glycine, histidine, isoleucine, leucine, lysine, provided that whena is one, the A ring is a heterocyclic 6 to methionine, phenylalanine, proline, serine, threonine, tryp 12-membered ring; and when the A ring is a heterocyclic tophan, tyrosine, Valine, homoarginine, homolysine, orni 5-membered ring, then a is an integer from 2 to 8; thine, arginine mimic, arginine homologue, arginine truncate, 35 each R is independently hydrogen, alkyl, substituted arginine with varying oxidation states, lysine mimic, lysine alkyl, aryl or substituted aryl; homologue, lysine truncate, and lysine with varying oxida R represents a side chain of an amino acid, a side chain of tion states; an amino acid variant, a derivative of a side chain of an amino each R is a side chain of an amino acid independently acid, or a derivative of a side chain of an amino acid variant selected from alanine, arginine, asparagine, aspartic acid, 40 that effects - C(O)-CH(R) N(R) to be a GI enzyme cysteine, glutamic acid, glutamine, glycine, histidine, isoleu cleavable moiety; cine, leucine, lysine, methionine, phenylalanine, proline, each R' represents a side chain of an amino acid indepen serine, threonine, tryptophan, tyrosine and Valine; dently selected from alanine, arginine, asparagine, aspartic b is a number from Zero to 100: 45 acid, cysteine, glutamic acid, glutamine, glycine, histidine, R’ is selected from hydrogen, alkyl, substituted alkyl, acyl, isoleucine, leucine, lysine, methionine, phenylalanine, pro Substituted acyl, alkoxycarbonyl, Substituted alkoxycarbo line, serine, threonine, tryptophan, tyrosine and valine; nyl, aryl, Substituted aryl, arylalkyl, and Substituted arylalkyl; b is a number from Zero to 100: or a salt, hydrate or solvate thereof. R’ is selected from hydrogen, alkyl, substituted alkyl, acyl, The present embodiments provide a compound of formula 50 Substituted acyl, alkoxycarbonyl, Substituted alkoxycarbo V: nyl, aryl, Substituted aryl, arylalkyl, and Substituted arylalkyl; or a salt, hydrate or solvate thereof. In formula II and IV-V, X can be a residue of a ketone (V) containing opioid. 55 A “ketone-containing opioid refers to a subset of the l R R2 O R3 R6 opioids that contain a ketone group. As used herein, a ketone R7 X N d N containing opioid is an opioid containing an enolizable N ketone group. A ketone-containing opioid is a compound with A a pharmacophore that presents to the opioid receptor an aro H R5 O R3 60 matic group and an aliphatic amine group in an architectur (Y), ally discrete way. See, for example, Foye's Principles of Medicinal Chemistry, Sixth Edition, ed. T. L. Lemke and D. wherein A. Williams, Lippincott Williams & Wilkins, 2008, particu X represents a residue of a ketone-containing opioid, 65 larly Chapter 24, pages 653-678. wherein the hydrogen atom of the corresponding hydroxyl For example, ketone-containing opioids include, but are group of the enolic tautomer of the ketone is replaced by a not limited to, acetylmorphone, hydrocodone, hydromor US 8,685,916 B2 27 28 phone, ketobemidone, methadone, naloxone, naltrexone, N-(oxycodone-6-enol-carbonyl)piperidine-2-methylamine N-methylmaloxone, N-methylmaltrexone, oxycodone, oxy L-arginine-L-alanine-acetate (Compound KC-13): morphone, and pentamorphone. In certain embodiments, the ketone-containing opioid is hydrocodone, hydromorphone, oxycodone, or oxymorphone. In certain embodiments, the ketone-containing opioid is naloxone, naltrexone, N-methylnaloxone, or N-methylmaltr CXO. In certain embodiments, the ketone-containing opioid is 10 hydrocodone or oxycodone. In certain embodiments, the ketone-containing opioid is hydrocodone. In certain embodi ments, the ketone-containing opioid is oxycodone. It is contemplated that opioids bearing at least some of the functionalities described herein will be developed; such opio 15 ids are included as part of the scope of this disclosure. In formula III, R' can be hydrogen or hydroxyl. In certain instances, R is hydrogen. In other instances, R is hydroxyl. Informula III, R is hydrogen or alkyl. In certain instances, R’ is hydrogen. In other instances, R is alkyl. Particular compounds of interest, and salts or Solvates or stereoisomers thereof, include: N-(oxycodone-6-enol-carbonyl)-R-(piperidine-2-methy lamine)-L-arginine-glycine-malonate (Compound N-(oxycodone-6-enol-carbonyl)piperidine-2-methylamine KC-17): L-arginine-glycine-acetate (Compound KC-14):

N-(oxycodone-6-enol-carbonyl)piperidine-2-methylamine L-arginine-malonate (Compound KC-12): 50

55

O O NH ls N 1N s 60 O NH

H2 -- H

65 N-(oxycodone-6-enol-carbonyl)piperidine-2-methylamine L-arginine-L-alanine-malonate (Compound KC-15): US 8,685,916 B2 30

N N O

NH

HN N

N-(oxycodone-6-enol-carbonyl)piperidine-2-methylamine L-arginine-glycine-malonate (Compound KC-16):

CH N 1. 3 OH o HC-O O N r O O O N N O H 8. d

NH

HN N

N-(hydrocodone-6-enol-carbonyl)-R-(piperidine-2-methy lamine)-L-arginine-glycine-malonate (Compound 45 KC-31):

US 8,685,916 B2 31 32 Particular compounds of interest, and salts or Solvates or Compound KC-37: stereoisomers thereof, include: Compound KC-32:

V N 3 O OH ( )N ) 10 - O O O N O O N^^ N J. 15 O NH

HN l N 2 H

Compound KC-35:

Compound KC-36: Compound KC-38:

50

55 E ls.O 60

65 US 8,685,916 B2 33 34 Compound KC-39: Compound KC-42:

Compound KC-43: Compound KC-40: 25

30

O

OH 35 O N-a-lE O H H SNY,O NY: 40 usesNH 2 H

45

Compound KC-44: Compound KC-41: 50

US 8,685,916 B2 35 36 Compound KC-45:

O O O NH lus N 1sN OH O NH

HN N

25 Compound KC-46: Compound KC-48:

30 Ols 35 s O O O N H ^s.H O

40 HN N 2 H

Compound KC-47:

O s

O O O NH lus N 1sN OH.; O

HN N US 8,685,916 B2 37 38 Compound KC-49: Compound KC-51:

O 10 lsOH OH s O O NN 15 H SN 1. O NH H -- HN 2 H r

Compound KC-50: Compound KC-52: 25

30 O

OH

O 35 OH. NN . N H O O

HN 40 HN

Compound KC-53:

OH

O O N

HN US 8,685,916 B2 39 40 Compound KC-55: stituted aminoacyl, amino, Substituted amino, acylamino, Substituted acylamino, and cyano;

c is a number from Zero to 3: each R' is independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, sub stituted alkynyl, aryl, Substituted aryl, acyl, Substituted acyl, O carboxyl, alkoxycarbonyl, Substituted alkoxycarbonyl, ami noacyl, Substituted aminoacyl, amino, Substituted amino, so acylamino, Substituted acylamino, and cyano; -O O O N 10 each R is independently selected from hydrogen, alkyl, Sry O substituted alkyl, alkenyl, substituted alkenyl, alkynyl, sub stituted alkynyl, aryl, Substituted aryl, acyl, Substituted acyl, NNH 1NH carboxyl, alkoxycarbonyl, Substituted alkoxycarbonyl, ami O y O ls noacyl, Substituted aminoacyl, amino, Substituted amino, 15 acylamino, Substituted acylamino, and cyano; or R" and R together with the carbon to which they are H.N. N attached can form a cycloalkyl or Substituted cycloalkyl group, or two R' or R groups on adjacent carbon atoms, Phenolic Opioid Prodrugs together with the carbonatoms to which they are attached, can The disclosure provides a phenolic opioid prodrug that form a cycloalkyl or Substituted cycloalkyl group; a is an integer from one to 8: provides controlled release of a phenolic opioid. In a phenolic provided that whena is one, the A ring is a heterocyclic 6 to opioid prodrug, a promoiety is attached to the phenolic opioid 12-membered ring; and when the A ring is a heterocyclic through the phenolic oxygen atom. In a phenolic opioid pro 5-membered ring, then a is an integer from 2 to 8; drug, the oxygen atom of the phenol group of the phenolic 25 each R is independently hydrogen, alkyl, substituted opioid is replaced by a covalent bond to a promoiety. alkyl, aryl or substituted aryl; As disclosed herein, an enzyme-cleavable phenolic opioid R is selected from hydrogen, alkyl, substituted alkyl, aryl, prodrug is a phenolic opioid prodrug that comprises a pro substituted aryl, arylalkyl, substituted arylalkyl, heteroalkyl, moiety comprising an enzyme-cleavable moiety, i.e., a moi substituted heteroalkyl, heteroaryl, substituted heteroaryl, ety having a site Susceptible to cleavage by an enzyme. In one 30 heteroarylalkyl, and substituted heteroarylalkyl: embodiment, the cleavable moiety is a GI enzyme-cleavable each R is independently selected from hydrogen, alkyl, moiety, such as a trypsin-cleavable moiety. Such a prodrug substituted alkyl, aryl, substituted aryl, arylalkyl, substituted comprises a phenolic opioid covalently bound to a promoiety arylalkyl, heteroalkyl, substituted heteroalkyl, heteroaryl, comprising an enzyme-cleavable moiety, wherein cleavage of substituted heteroaryl, heteroarylalkyl, and substituted het the enzyme-cleavable moiety by an enzyme mediates release 35 eroarylalkyl: of the drug. b is a number from Zero to 100: Formulae VI-IX R’ is selected from hydrogen, alkyl, substituted alkyl, acyl, Compounds of the present disclosure include compounds Substituted acyl, alkoxycarbonyl, Substituted alkoxycarbo of formulae VI-IX shown below. Compositions of the present nyl, aryl, Substituted aryl, arylalkyl, and Substituted arylalkyl; disclosure also include compounds of formulae VI-IX shown 40 or a salt, hydrate or solvate thereof. below. Pharmaceutical compositions and methods of the The present embodiments provide a compound of formula present disclosure also contemplate compounds of formulae VII: VI-IX. The present embodiments provide a compound of formula VI: 45 (VII)

50 O R1 R2 O 6 N R7

(Y), A H. R. LÖ RJ, 55 (Y), wherein X represents a residue of a phenolic opioid, wherein the wherein hydrogenatom of the phenolic hydroxyl group is replaced by R" is hydrogen or hydroxyl; a covalent bond to C(O) N (A ring)-Y. (CRR), 60 R’ is hydrogen or alkyl; NH C(O) CH(R) N(R) C(O) CH(R) the A ring is a heterocyclic 5 to 12-membered ring: N(R) R7: each Y is independently selected from alkyl, substituted the A ring is a heterocyclic 5 to 12-membered ring; alkyl, alkenyl, Substituted alkenyl, alkynyl, Substituted alky each Y is independently selected from alkyl, substituted nyl, aryl, Substituted aryl, acyl, Substituted acyl, carboxyl, alkyl, alkenyl, Substituted alkenyl, alkynyl, Substituted alky 65 alkoxycarbonyl, Substituted alkoxycarbonyl, acylamino, Sub nyl, aryl, Substituted aryl, acyl, Substituted acyl, carboxyl, stituted acylamino, Substituted aminoacyl, amino, Substituted alkoxycarbonyl. Substituted alkoxycarbonyl, aminoacyl, Sub amino, acylamino, and cyano; US 8,685,916 B2 41 42 c is a number from Zero to 3: each R' is independently selected from hydrogen, alkyl, each R" is independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, sub substituted alkyl, alkenyl, substituted alkenyl, alkynyl, sub stituted alkynyl, aryl, Substituted aryl, acyl, Substituted acyl, stituted alkynyl, aryl, Substituted aryl, acyl, Substituted acyl, carboxyl, alkoxycarbonyl, Substituted alkoxycarbonyl, ami carboxyl, alkoxycarbonyl, Substituted alkoxycarbonyl, ami noacyl, Substituted aminoacyl, amino, Substituted amino, noacyl, Substituted aminoacyl, amino, Substituted amino, acylamino, Substituted acylamino, and cyano; acylamino, Substituted acylamino, and cyano; each R is independently selected from hydrogen, alkyl, each R is independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, sub substituted alkyl, alkenyl, substituted alkenyl, alkynyl, sub stituted alkynyl, aryl, Substituted aryl, acyl, Substituted acyl, stituted alkynyl, aryl, Substituted aryl, acyl, Substituted acyl, 10 carboxyl, alkoxycarbonyl, Substituted alkoxycarbonyl, ami carboxyl, alkoxycarbonyl, Substituted alkoxycarbonyl, ami noacyl, Substituted aminoacyl, amino, Substituted amino, noacyl, Substituted aminoacyl, amino, Substituted amino, acylamino, Substituted acylamino, and cyano; or acylamino, Substituted acylamino, and cyano; or R" and R together with the carbon to which they are R" and R together with the carbon to which they are 15 attached can form a cycloalkyl or Substituted cycloalkyl attached can form a cycloalkyl or Substituted cycloalkyl group, or two R' or R groups on adjacent carbon atoms, group, or two R' or R groups on adjacent carbon atoms, together with the carbonatoms to which they are attached, can together with the carbonatoms to which they are attached, can form a cycloalkyl or Substituted cycloalkyl group; form a cycloalkyl or Substituted cycloalkyl group; a is an integer from one to 8: a is an integer from one to 8: provided that whena is one, the A ring is a heterocyclic 6 to provided that whena is one, the Aring is a heterocyclic 6 to 12-membered ring; and when the A ring is a heterocyclic 12-membered ring; and when the A ring is a heterocyclic 5-membered ring, then a is an integer from 2 to 8; 5-membered ring, then a is an integer from 2 to 8: each R is independently hydrogen, alkyl, substituted each R is independently hydrogen, alkyl, substituted alkyl, aryl or substituted aryl; alkyl, aryl or substituted aryl; 25 R is a side chain of an amino acid selected from alanine, R is selected from hydrogen, alkyl, substituted alkyl, aryl, arginine, asparagine, aspartic acid, cysteine, glutamic acid, substituted aryl, arylalkyl, substituted arylalkyl, heteroalkyl, glutamine, glycine, histidine, isoleucine, leucine, lysine, substituted heteroalkyl, heteroaryl, substituted heteroaryl, methionine, phenylalanine, proline, serine, threonine, tryp heteroarylalkyl, and substituted heteroarylalkyl; tophan, tyrosine, Valine, homoarginine, homolysine, orni each R is independently selected from hydrogen, alkyl, 30 thine, arginine mimic, arginine homologue, arginine truncate, substituted alkyl, aryl, substituted aryl, arylalkyl, substituted arginine with varying oxidation states, lysine mimic, lysine arylalkyl, heteroalkyl, substituted heteroalkyl, heteroaryl, homologue, lysine truncate, and lysine with varying oxida substituted heteroaryl, heteroarylalkyl, and substituted het tion states; eroarylalkyl: each R is a side chain of an amino acid independently b is a number from Zero to 100: 35 selected from alanine, arginine, asparagine, aspartic acid, R’ is selected from hydrogen, alkyl, substituted alkyl, acyl, cysteine, glutamic acid, glutamine, glycine, histidine, isoleu Substituted acyl, alkoxycarbonyl, Substituted alkoxycarbo cine, leucine, lysine, methionine, phenylalanine, proline, nyl, aryl, Substituted aryl, arylalkyl, and Substituted arylalkyl; serine, threonine, tryptophan, tyrosine and Valine; or a salt, hydrate or solvate thereof. b is a number from Zero to 100: The present embodiments provide a compound of formula 40 R’ is selected from hydrogen, alkyl, substituted alkyl, acyl, VIII: Substituted acyl, alkoxycarbonyl, Substituted alkoxycarbo nyl, aryl, Substituted aryl, arylalkyl, and Substituted arylalkyl; or a salt, hydrate or solvate thereof. (VIII) The present embodiments provide a compound of formula 45 IX: O R R2 Q R3 R6 l N R7 X N a N N A H R5 O RJ, 50 (Y),

wherein (Y), X represents a residue of a phenolic opioid, wherein the 55 hydrogenatom of the phenolic hydroxyl group is replaced by a covalent bond to C(O) N (A ring)-Y. (CRR), wherein NH C(O) CH(R) N(R) C(O) CH(R) X represents a residue of a phenolic opioid, wherein the N(R) R7: hydrogenatom of the phenolic hydroxyl group is replaced by the A ring is a heterocyclic 5 to 12-membered ring; 60 a covalent bond to —C(O) N (A ring)-Y. (CRR), each Y is independently selected from alkyl, substituted NH CO) CH(R) N(R) C(O) CH(R) alkyl, alkenyl, Substituted alkenyl, alkynyl, Substituted alky N(R) R7: nyl, aryl, Substituted aryl, acyl, Substituted acyl, carboxyl, the A ring is a heterocyclic 5 to 12-membered ring: alkoxycarbonyl. Substituted alkoxycarbonyl, aminoacyl, Sub each Y is independently selected from alkyl, substituted stituted aminoacyl, amino, Substituted amino, acylamino, 65 alkyl, alkenyl, Substituted alkenyl, alkynyl, Substituted alky Substituted acylamino, and cyano; nyl, aryl, Substituted aryl, acyl, Substituted acyl, carboxyl, c is a number from Zero to 3: alkoxycarbonyl, Substituted alkoxycarbonyl, aminoacyl, Sub US 8,685,916 B2 43 44 stituted aminoacyl, amino, Substituted amino, acylamino, In certain embodiments, the phenolic opioid is naloxone, Substituted acylamino, and cyano; maltrexone, N-methylnaloxone, or N-methylmaltrexone. In c is a number from Zero to 3: certain embodiments, the phenolic opioid is diprenorphine or each R" is independently selected from hydrogen, alkyl, N-methyldiprenorphine. substituted alkyl, alkenyl, substituted alkenyl, alkynyl, sub In certain embodiments, the phenolic opioid is hydromor stituted alkynyl, aryl, Substituted aryl, acyl, Substituted acyl, phone. In certain embodiments, the phenolic opioid is mor carboxyl, alkoxycarbonyl, Substituted alkoxycarbonyl, ami phine. In certain embodiments, the phenolic opioid is oxy noacyl, Substituted aminoacyl, amino, Substituted amino, morphone. In certain embodiments, the phenolic opioid is acylamino, Substituted acylamino, and cyano; 10 tapentadol. each R is independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, sub It is contemplated that opioids bearing at least some of the stituted alkynyl, aryl, Substituted aryl, acyl, Substituted acyl, functionalities described herein will be developed; such opio carboxyl, alkoxycarbonyl, Substituted alkoxycarbonyl, ami ids are included as part of the scope of this disclosure. noacyl, Substituted aminoacyl, amino, Substituted amino, 15 Informula VII, R' can be hydrogen or hydroxyl. In certain acylamino, Substituted acylamino, and cyano; or instances, R' is hydrogen. In other instances, R is hydroxyl. R" and R together with the carbon to which they are In formula VII, R is hydrogen or alkyl. In certain attached can form a cycloalkyl or Substituted cycloalkyl instances, R is hydrogen. In other instances, R is alkyl. group, or two R' or R groups on adjacent carbon atoms, Particular compound of interest, and salts or Solvates or together with the carbonatoms to which they are attached, can Stereoisomers thereof, includes: form a cycloalkyl or Substituted cycloalkyl group; N-(Tapentadol-carbonyl)piperidine-2-methylamine-L-argi a is an integer from one to 8: nine-malonate (Compound TP-5): provided that whena is one, the Aring is a heterocyclic 6 to 12-membered ring; and when the A ring is a heterocyclic 25 5-membered ring, then a is an integer from 2 to 8: H each R is independently hydrogen, alkyl, substituted "N." alkyl, aryl or substituted aryl; N NH R represents a side chain of an amino acid, a side chain of 30 --- N l O O an amino acid variant, a derivative of a side chain of an amino N acid, or a derivative of a side chain of an amino acid variant O H N lus OH. that effects—C(O)-CH(R) N(R) to be a GI enzyme H cleavable moiety; Ol O ullN each R represents a side chain of an amino acid indepen 35 dently selected from alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, pro line, serine, threonine, tryptophan, tyrosine and valine; Amide-modified Opioid Prodrugs b is a number from Zero to 100: 40 The disclosure provides an amide-modified opioid prodrug R’ is selected from hydrogen, alkyl, substituted alkyl, acyl, that provides controlled release of an amide-containing Substituted acyl, alkoxycarbonyl, Substituted alkoxycarbo opioid. As shown below, in an amide-modified opioid pro nyl, aryl, Substituted aryl, arylalkyl, and Substituted arylalkyl; drug, a promoiety is attached to the amide-containing opioid or a salt, hydrate or solvate thereof. 45 through the enolic oxygen atom of the amide enol moiety or Informula VI and VIII-IX, X can be a residue of a phenolic through the oxygen of the imine tautomer. In an amide-modi opioid. fied opioid prodrug, the hydrogen atom of the corresponding A "phenolic opioid refers to a subset of the opioids that enolic group of the amide enol or of the imine tautomer of the contain a phenol group. A phenolic opioid is a compound with amide-containing opioid is replaced by a covalent bond to a a pharmacophore that presents to the opioid receptor an aro 50 promoiety. In certain embodiments, the promoiety that matic group and an aliphatic amine group in an architectur replaces the hydrogen atom of the corresponding enolic ally discrete way. See, for example, Foye's Principles of group of the amide enol or the imine tautomer of the amide Medicinal Chemistry, Sixth Edition, ed. T. L. Lemke and D. containing opioid contains an acyl group as the point of A. Williams, Lippincott Williams & Wilkins, 2008, particu connection. larly Chapter 24, pages 653-678. 55 For instance, the following opioids contain a phenol group that can be a point of attachment to a promoiety: buprenor O R R2 Q R3 R6 phine, dihydroetorphine, diprenorphine, etorphine, hydro l N R7 O N a N N morphone, levorphanol, morphine, nalbuphine, nalmefene, 60 A nalorphine, naloxone, naltrexone, N-methyldiprenorphine, H R5 O R3 N-methylmaloxone, N-methylmaltrexone, oripavine, oxymor R NR phone, butorphanol, dezocine, ketobemidone, meptazinol, (Y), o-desmethyltramadol, pentazocine, phenazocine, and tapen imine tautomer tadol. 65 where RandR are collectively the rest of the amide-containing opioid or one of R and In certain embodiments, the phenolic opioid is hydromor R’ is hydrogen and the other is the rest is the amide-containing opioid phone, morphine, oxymorphone, or tapentadol. US 8,685,916 B2 46 -continued noacyl, Substituted aminoacyl, amino, Substituted amino, O R1 R2 Q R6 acylamino, Substituted acylamino, and cyano; or ls N R7 R" and R together with the carbon to which they are O N a N N attached can form a cycloalkyl or Substituted cycloalkyl 5 A group, or two R' or R groups on adjacent carbon atoms, H R5 O R3 together with the carbonatoms to which they are attached, can R NRR" form a cycloalkyl or Substituted cycloalkyl group; (Y), a is an integer from one to 8: amide enol provided that whena is one, the A ring is a heterocyclic 6 to where R, RandR" are collectively the rest of the amide-containing opioid or at least 10 12-membered ring; and when the A ring is a heterocyclic one of R, RandR" is hydrogen and the rest of the R, RandR" are collectively the rest 5-membered ring, then a is an integer from 2 to 8; of the amide-containing opioid each R is independently hydrogen, alkyl, substituted alkyl, aryl or substituted aryl; As disclosed herein, an enzyme-cleavable amide-modified R is selected from hydrogen, alkyl, substituted alkyl, aryl, opioid prodrug is an amide-modified opioid prodrug that 15 substituted aryl, arylalkyl, substituted arylalkyl, heteroalkyl, comprises a promoiety comprising an enzyme-cleavable substituted heteroalkyl, heteroaryl, substituted heteroaryl, moiety, i.e., a moiety having a site Susceptible to cleavage by heteroarylalkyl, and substituted heteroarylalkyl: an enzyme. Release of the opioid is mediated by enzymatic each R is independently selected from hydrogen, alkyl, cleavage of the promoiety from the amide-containing opioid. substituted alkyl, aryl, substituted aryl, arylalkyl, substituted In one embodiment, the cleavable moiety is a GI enzyme arylalkyl, heteroalkyl, substituted heteroalkyl, heteroaryl, cleavable moiety, such as a trypsin-cleavable moiety. substituted heteroaryl, heteroarylalkyl, and substituted het Formulae X-XII eroarylalkyl: b is a number from Zero to 100: Compounds of the present disclosure include compounds R’ is selected from hydrogen, alkyl, substituted alkyl, acyl, of formulae X-XII shown below. Compositions of the present 25 Substituted acyl, alkoxycarbonyl, Substituted alkoxycarbo disclosure also include compounds of formulae X-XII shown nyl, aryl, Substituted aryl, arylalkyl, and Substituted arylalkyl; below. Pharmaceutical compositions and methods of the or a salt, hydrate or solvate thereof. present disclosure also contemplate compounds of formulae The present embodiments provide a compound of formula X-XII. XI: The present embodiments provide a compound of formula 30 X: (XI) O R R2 Q R3 R6 (X) O R R2 Q R3 R6 l N R7 35 X N a N N A l N R7 H R5 O R3 x -A (- - H R5 O RJ, (Y), (Y), 40 wherein X represents a residue of an amide-containing opioid, wherein wherein –C(O) N(A ring)-Y-(CRR). NH C X represents a residue of an amide-containing opioid, (O) CH(R) N(R) C(O) CH(R) N(R) R7 is wherein –C(O) N(A ring)-Y-(CRR). NH C 45 connected to the amide-containing opioid through the oxygen (O) CH(R) N(R) C(O) CH(R) N(R) R7 is of the amide group, wherein the amide group is converted to connected to the amide-containing opioid through the oxygen an amide enol or an imine tautomer, of the amide group, wherein the amide group is converted to the A ring is a heterocyclic 5 to 12-membered ring: an amide enol or an imine tautomer, each Y is independently selected from alkyl, substituted the A ring is a heterocyclic 5 to 12-membered ring; 50 alkyl, alkenyl, Substituted alkenyl, alkynyl, Substituted alky each Y is independently selected from alkyl, substituted nyl, aryl, Substituted aryl, acyl, Substituted acyl, carboxyl, alkyl, alkenyl, Substituted alkenyl, alkynyl, Substituted alky alkoxycarbonyl, Substituted alkoxycarbonyl, aminoacyl, Sub nyl, aryl, Substituted aryl, acyl, Substituted acyl, carboxyl, stituted aminoacyl, amino, Substituted amino, acylamino, alkoxycarbonyl. Substituted alkoxycarbonyl, aminoacyl, Sub Substituted acylamino, and cyano; stituted aminoacyl, amino, Substituted amino, acylamino, 55 c is a number from Zero to 3: Substituted acylamino, and cyano; each R' is independently selected from hydrogen, alkyl, c is a number from Zero to 3: substituted alkyl, alkenyl, substituted alkenyl, alkynyl, sub each R" is independently selected from hydrogen, alkyl, stituted alkynyl, aryl, Substituted aryl, acyl, Substituted acyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, sub carboxyl, alkoxycarbonyl, Substituted alkoxycarbonyl, ami stituted alkynyl, aryl, Substituted aryl, acyl, Substituted acyl, 60 noacyl, Substituted aminoacyl, amino, Substituted amino, carboxyl, alkoxycarbonyl, Substituted alkoxycarbonyl, ami acylamino, Substituted acylamino, and cyano; noacyl, Substituted aminoacyl, amino, Substituted amino, each R is independently selected from hydrogen, alkyl, acylamino, Substituted acylamino, and cyano; substituted alkyl, alkenyl, substituted alkenyl, alkynyl, sub each R is independently selected from hydrogen, alkyl, stituted alkynyl, aryl, Substituted aryl, acyl, Substituted acyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, sub 65 carboxyl, alkoxycarbonyl, Substituted alkoxycarbonyl, ami stituted alkynyl, aryl, Substituted aryl, acyl, Substituted acyl, noacyl, Substituted aminoacyl, amino, Substituted amino, carboxyl, alkoxycarbonyl, Substituted alkoxycarbonyl, ami acylamino, Substituted acylamino, and cyano; or US 8,685,916 B2 47 48 R" and R together with the carbon to which they are each R is independently selected from hydrogen, alkyl, attached can form a cycloalkyl or Substituted cycloalkyl substituted alkyl, alkenyl, substituted alkenyl, alkynyl, sub group, or two R' or R groups on adjacent carbon atoms, stituted alkynyl, aryl, Substituted aryl, acyl, Substituted acyl, together with the carbonatoms to which they are attached, can carboxyl, alkoxycarbonyl, Substituted alkoxycarbonyl, ami form a cycloalkyl or Substituted cycloalkyl group; noacyl, Substituted aminoacyl, amino, Substituted amino, a is an integer from one to 8: acylamino, Substituted acylamino, and cyano; or provided that whena is one, the Aring is a heterocyclic 6 to R" and R together with the carbon to which they are 12-membered ring; and when the A ring is a heterocyclic attached can form a cycloalkyl or Substituted cycloalkyl 5-membered ring, then a is an integer from 2 to 8: 10 group, or two R' or R groups on adjacent carbon atoms, each R is independently hydrogen, alkyl, substituted together with the carbonatoms to which they are attached, can alkyl, aryl or substituted aryl; form a cycloalkyl or Substituted cycloalkyl group; R is a side chain of an amino acid selected from alanine, a is an integer from one to 8: arginine, asparagine, aspartic acid, cysteine, glutamic acid, provided that whena is one, the A ring is a heterocyclic 6 to glutamine, glycine, histidine, isoleucine, leucine, lysine, 15 12-membered ring; and when the A ring is a heterocyclic methionine, phenylalanine, proline, serine, threonine, tryp 5-membered ring, then a is an integer from 2 to 8; tophan, tyrosine, Valine, homoarginine, homolysine, orni each R is independently hydrogen, alkyl, substituted thine, arginine mimic, arginine homologue, arginine truncate, alkyl, aryl or substituted aryl; arginine with varying oxidation states, lysine mimic, lysine R represents a side chain of an amino acid, a side chain of homologue, lysine truncate, and lysine with varying oxida an amino acid variant, a derivative of a side chain of an amino tion states; acid, or a derivative of a side chain of an amino acid variant each R is a side chain of an amino acid independently that effects - C(O)-CH(R) N(R) to be a GI enzyme selected from alanine, arginine, asparagine, aspartic acid, cleavable moiety; cysteine, glutamic acid, glutamine, glycine, histidine, isoleu 25 cine, leucine, lysine, methionine, phenylalanine, proline, each R' represents a side chain of an amino acid indepen serine, threonine, tryptophan, tyrosine and Valine; dently selected from alanine, arginine, asparagine, aspartic b is a number from Zero to 100: acid, cysteine, glutamic acid, glutamine, glycine, histidine, R’ is selected from hydrogen, alkyl, substituted alkyl, acyl, isoleucine, leucine, lysine, methionine, phenylalanine, pro Substituted acyl, alkoxycarbonyl, Substituted alkoxycarbo 30 line, serine, threonine, tryptophan, tyrosine and valine; nyl, aryl, Substituted aryl, arylalkyl, and Substituted arylalkyl; b is a number from Zero to 100: or a salt, hydrate or solvate thereof. R’ is selected from hydrogen, alkyl, substituted alkyl, acyl, The present embodiments provide a compound of formula Substituted acyl, alkoxycarbonyl, Substituted alkoxycarbo XII: nyl, aryl, Substituted aryl, arylalkyl, and Substituted arylalkyl; 35 or a salt, hydrate or solvate thereof. Informula X-XII, X can be a residue of an amide-contain ing opioid, where the amide-containing opioid is connected through the oxygen of the amide group, wherein the amide group is converted to an amide enol or an imine tautomer. 40 An "amide-containing opioid refers to a subset of the opioids that contain an amide group. As used herein, an amide-containing opioid is an opioid containing an enoliz (Y), able amide group. An amide-containing opioid is a compound 45 with a pharmacophore that presents to the opioid receptor an wherein aromatic group and an aliphatic amine group in an architec X represents a residue of an amide-containing opioid, turally discrete way. See, for example, Foye's Principles of wherein –C(O) N(A ring)-Y-(CRR). NH C Medicinal Chemistry, Sixth Edition, ed. T. L. Lemke and D. (O) CH(R) N(R) C(O) CH(R) N(R) R7 is A. Williams, Lippincott Williams & Wilkins, 2008, particu connected to the amide-containing opioid through the oxygen 50 larly Chapter 24, pages 653-678. of the amide group, wherein the amide group is converted to For instance, the following opioids contain an amide group an amide enol or an imine tautomer, that can be a point of attachment to a promoiety: alfentanil, the A ring is a heterocyclic 5 to 12-membered ring; carfentanil, fentanyl, lofentanil, loperamide, olmefentanyl. each Y is independently selected from alkyl, substituted alkyl, alkenyl, Substituted alkenyl, alkynyl, Substituted alky 55 remifentanil, and Sufentanil. nyl, aryl, Substituted aryl, acyl, Substituted acyl, carboxyl, It is contemplated that opioids bearing at least some of the alkoxycarbonyl. Substituted alkoxycarbonyl, aminoacyl, Sub functionalities described herein will be developed; such opio stituted aminoacyl, amino, Substituted amino, acylamino, ids are included as part of the scope of this disclosure. Compounds with Certain A Rings Substituted acylamino, and cyano; 60 c is a number from Zero to 3: Formulae XIII-XV each R" is independently selected from hydrogen, alkyl, Compounds of the present disclosure include compounds substituted alkyl, alkenyl, substituted alkenyl, alkynyl, sub of formulae XIII-XV shown below. Compositions of the stituted alkynyl, aryl, Substituted aryl, acyl, Substituted acyl, present disclosure also include compounds of formulae XIII carboxyl, alkoxycarbonyl, Substituted alkoxycarbonyl, ami 65 XV shown below. Pharmaceutical compositions and methods noacyl, Substituted aminoacyl, amino, Substituted amino, of the present disclosure also contemplate compounds of acylamino, Substituted acylamino, and cyano; formulae XIII-XV. US 8,685,916 B2 49 50 The present embodiments provide a compound of formula b is a number from Zero to 100; and XIII: R’ is selected from hydrogen, alkyl, substituted alkyl, acyl, Substituted acyl, alkoxycarbonyl, Substituted alkoxycarbo nyl, aryl, Substituted aryl, arylalkyl, and Substituted arylalkyl; (XIII) or a salt, hydrate or solvate thereof. O R R2 Q R3 R6 The present embodiments provide a compound of formula XIV: ls A. N R7 X N1 ><2 s k A k H R5 O RJ, 10 (XIV) NA41. Ny O R1 R2 O R3 R6 R7 wherein X l N 4-4.A. N N N X is selected from a residue of a ketone-containing opioid, 15 A wherein the hydrogen atom of the corresponding hydroxyl A NA41.A Ny H R5 O RJ,3 group of the enolic tautomer of the ketone is replaced by a covalent bond to C(O) N (A ring)-Y (CRR), NH C(O) CH(R) N(R) C(O) CH(R) wherein N(R) R'; a residue of a phenolic opioid, wherein the X is selected from a residue of a ketone-containing opioid, hydrogenatom of the phenolic hydroxyl group is replaced by wherein the hydrogen atom of the corresponding hydroxyl a covalent bond to C(O) N (A ring)-Y (CRR), group of the enolic tautomer of the ketone is replaced by a NH C(O) CH(R) N(R) C(O) CH(R) covalent bond to C(O) N(A ring)-Y-(CRR) N(R) R', and a residue of an amide-containing opioid, NH CO) CH(R) N(R) C(O) CH(R) wherein –C(O) NI (A ring)-Y-(CRR). NH C 25 N(R) R'; a residue of a phenolic opioid, wherein the (O) CH(R) N(R) C(O) CH(R) N(R) R7 is hydrogenatom of the phenolic hydroxyl group is replaced by connected to the amide-containing opioid through the oxygen a covalent bond to C(O) N (A ring)-Y CRR), of the amide group, wherein the amide group is converted to NH CO) CH(R) N(R) C(O) CH(R) an amide enol or an imine tautomer, N(R) R', and a residue of an amide-containing opioid, A'. A. A. and Aare independently selected from carbon, 30 wherein –C(O) N(A ring)-Y-(CRR). NH C nitrogen, oxygen, and Sulfur, (O) CH(R) N(R) C(O) CH(R) N(R) R7 is A is carbon or nitrogen; connected to the amide-containing opioid through the oxygen each Y is independently selected from alkyl, substituted of the amide group, wherein the amide group is converted to alkyl, alkenyl, Substituted alkenyl, alkynyl, Substituted alky an amide enol or an imine tautomer, nyl, aryl, Substituted aryl, acyl, Substituted acyl, carboxyl, 35 A'. A. A. and Aare independently selected from carbon, alkoxycarbonyl. Substituted alkoxycarbonyl, aminoacyl, Sub nitrogen, oxygen, and Sulfur, stituted aminoacyl, amino, Substituted amino, acylamino, A is carbon or nitrogen; Substituted acylamino, and cyano; Y is selected from alkyl, substituted alkyl, alkenyl, substi each R" is independently selected from hydrogen, alkyl, tuted alkenyl, alkynyl, Substituted alkynyl, aryl, Substituted substituted alkyl, alkenyl, substituted alkenyl, alkynyl, sub 40 aryl, acyl, Substituted acyl, carboxyl, alkoxycarbonyl, Substi stituted alkynyl, aryl, Substituted aryl, acyl, Substituted acyl, tuted alkoxycarbonyl, aminoacyl, Substituted aminoacyl, carboxyl, alkoxycarbonyl, Substituted alkoxycarbonyl, ami amino, Substituted amino, acylamino, Substituted acylamino, noacyl, Substituted aminoacyl, amino, Substituted amino, and cyano; acylamino, Substituted acylamino, and cyano; each R' is independently selected from hydrogen, alkyl, each R is independently selected from hydrogen, alkyl, 45 substituted alkyl, alkenyl, substituted alkenyl, alkynyl, sub substituted alkyl, alkenyl, substituted alkenyl, alkynyl, sub stituted alkynyl, aryl, Substituted aryl, acyl, Substituted acyl, stituted alkynyl, aryl, Substituted aryl, acyl, Substituted acyl, carboxyl, alkoxycarbonyl, Substituted alkoxycarbonyl, ami carboxyl, alkoxycarbonyl, Substituted alkoxycarbonyl, ami noacyl, Substituted aminoacyl, amino, Substituted amino, noacyl, Substituted aminoacyl, amino, Substituted amino, acylamino, Substituted acylamino, and cyano; acylamino, Substituted acylamino, and cyano; or 50 each R is independently selected from hydrogen, alkyl, R" and R together with the carbon to which they are substituted alkyl, alkenyl, substituted alkenyl, alkynyl, sub attached can form a cycloalkyl or Substituted cycloalkyl stituted alkynyl, aryl, Substituted aryl, acyl, Substituted acyl, group, or two R' or R groups on adjacent carbon atoms, carboxyl, alkoxycarbonyl, Substituted alkoxycarbonyl, ami together with the carbonatoms to which they are attached, can noacyl, Substituted aminoacyl, amino, Substituted amino, form a cycloalkyl or Substituted cycloalkyl group; 55 acylamino, Substituted acylamino, and cyano; or a is an integer from one to 8: R" and R together with the carbon to which they are each R is independently hydrogen, alkyl, substituted attached can form a cycloalkyl or Substituted cycloalkyl alkyl, aryl or substituted aryl; group, or two R' or R groups on adjacent carbon atoms, R is selected from hydrogen, alkyl, substituted alkyl, aryl, together with the carbonatoms to which they are attached, can substituted aryl, arylalkyl, substituted arylalkyl, heteroalkyl, 60 form a cycloalkyl or Substituted cycloalkyl group; substituted heteroalkyl, heteroaryl, substituted heteroaryl, a is an integer from one to 8: heteroarylalkyl, and substituted heteroarylalkyl; each R is independently hydrogen, alkyl, substituted each R is independently selected from hydrogen, alkyl, alkyl, aryl or substituted aryl; substituted alkyl, aryl, substituted aryl, arylalkyl, substituted R is a side chain of an amino acid selected from alanine, arylalkyl, heteroalkyl, substituted heteroalkyl, heteroaryl, 65 arginine, asparagine, aspartic acid, cysteine, glutamic acid, substituted heteroaryl, heteroarylalkyl, and substituted het glutamine, glycine, histidine, isoleucine, leucine, lysine, eroarylalkyl: methionine, phenylalanine, proline, serine, threonine, tryp US 8,685,916 B2 51 52 tophan, tyrosine, Valine, homoarginine, homolysine, orni group, or two R' or R groups on adjacent carbon atoms, thine, arginine mimic, arginine homologue, arginine truncate, together with the carbonatoms to which they are attached, can arginine with varying oxidation states, lysine mimic, lysine form a cycloalkyl or Substituted cycloalkyl group; homologue, lysine truncate, and lysine with varying oxida a is an integer from one to 8: tion states; each R is independently hydrogen, alkyl, substituted each R is a side chain of an amino acid independently alkyl, aryl or substituted aryl; selected from alanine, arginine, asparagine, aspartic acid, R represents a side chain of an amino acid, a side chain of cysteine, glutamic acid, glutamine, glycine, histidine, isoleu an amino acid variant, a derivative of a side chain of an amino cine, leucine, lysine, methionine, phenylalanine, proline, acid, or a derivative of a side chain of an amino acid variant serine, threonine, tryptophan, tyrosine and Valine; 10 that effects –C(O)-CH(R) N(R) to be a GI enzyme b is a number from Zero to 100: cleavable moiety; R’ is selected from hydrogen, alkyl, substituted alkyl, acyl, each R represents a side chain of an amino acid indepen Substituted acyl, alkoxycarbonyl, Substituted alkoxycarbo dently selected from alanine, arginine, asparagine, aspartic nyl, aryl, Substituted aryl, arylalkyl, and Substituted arylalkyl; acid, cysteine, glutamic acid, glutamine, glycine, histidine, or a salt, hydrate or solvate thereof. 15 isoleucine, leucine, lysine, methionine, phenylalanine, pro The present embodiments provide a compound of formula line, serine, threonine, tryptophan, tyrosine and valine; b is a number from Zero to 100: XV: R’ is selected from hydrogen, alkyl, substituted alkyl, acyl, Substituted acyl, alkoxycarbonyl, Substituted alkoxycarbo nyl, aryl, Substituted aryl, arylalkyl, and Substituted arylalkyl; (XV) or a salt, hydrate or solvate thereof. O R R2 O R3 R6 Informulae XIII-XV. X can be selected from a residue of a R7 ketone-containing opioid, X ls N 4-4.A. N N N wherein the hydrogenatom of the corresponding hydroxyl A 25 group of the enolic tautomer of the ketone is replaced by a AN-AN. H R5 O R3 covalent bond to C(O) N(A ring)-Y-(CRR) A4 Y NH CO) CH(R) N(R) C(O) CH(R) N(R) R'; a residue of a phenolic opioid, wherein the wherein hydrogenatom of the phenolic hydroxyl group is replaced by X is selected from a residue of a ketone-containing opioid, 30 a covalent bond to C(O) N(A ring)-Y-(CRR) wherein the hydrogen atom of the corresponding hydroxyl NH CO) CH(R) N(R) C(O) CH(R) group of the enolic tautomer of the ketone is replaced by a N(R) R'; and a residue of an amide-containing opioid, covalent bond to C(O) N(A ring)-Y (CRR) wherein –C(O) N(A ring)-Y-(CRR). NH C NH C(O) CH(R) N(R) C(O) CH(R) (O) CH(R) N(R) C(O) CH(R) N(R) R7 is N(R) R'; a residue of a phenolic opioid, wherein the 35 connected to the amide-containing opioid through the oxygen hydrogenatom of the phenolic hydroxyl group is replaced by of the amide group, wherein the amide group is converted to a covalent bond to C(O) N (A ring)-Y (CRR), an amide enol or an imine tautomer. NH C(O) CH(R) N(R) C(O) CH(R) In certain instances, X is a ketone-containing opioid, N(R) R', and a residue of an amide-containing opioid, wherein the hydrogen atom of the corresponding hydroxyl wherein –C(O) NI (A ring)-Y-(CRR). NH C 40 group of the enolic tautomer of the ketone is replaced by a (O) CH(R) N(R) C(O) CH(R) N(R) R7 is covalent bond to C(O) NI (A ring)-Y-(CRR), connected to the amide-containing opioid through the oxygen NH CO) CH(R) N(R) C(O) CH(R) of the amide group, wherein the amide group is converted to N(R) R7. an amide enol or an imine tautomer, In certain instances, X is a ketone-containing opioid, A'. A. A. and Aare independently selected from carbon, 45 wherein the opioid is selected from acetylmorphone, hydro nitrogen, oxygen, and Sulfur, codone, hydromorphone, ketobemidone, methadone, nalox A is carbon or nitrogen; one, naltrexone, N-methylnaloxone, N-methylmaltrexone, Y is selected from alkyl, substituted alkyl, alkenyl, substi oxycodone, oxymorphone, and pentamorphone. tuted alkenyl, alkynyl, Substituted alkynyl, aryl, Substituted In certain instances, X is a residue of a phenolic opioid, aryl, acyl, Substituted acyl, carboxyl, alkoxycarbonyl, Substi 50 wherein the hydrogenatom of the phenolic hydroxyl group is tuted alkoxycarbonyl, aminoacyl, Substituted aminoacyl, replaced by a covalent bond to —C(O) NI (A ring)-Y— amino, Substituted amino, acylamino, Substituted acylamino, (CRR). NH CO) CH(R) N(R) C(O) CH and cyano; (R) N(R) R7. each R" is independently selected from hydrogen, alkyl, In certain instances, X is a phenolic opioid, wherein the substituted alkyl, alkenyl, substituted alkenyl, alkynyl, sub 55 opioid is selected from buprenorphine, dihydroetorphine, stituted alkynyl, aryl, Substituted aryl, acyl, Substituted acyl, diprenorphine, etorphine, hydromorphone, levorphanol, carboxyl, alkoxycarbonyl, Substituted alkoxycarbonyl, ami morphine, nalbuphine, nalmefene, nalorphine, naloxone, nal noacyl, Substituted aminoacyl, amino, Substituted amino, trexone, N-methyldiprenorphine, N-methylnaloxone, N-me acylamino, Substituted acylamino, and cyano; thylmaltrexone, oripavine, oxymorphone, butorphanol, dezo each R is independently selected from hydrogen, alkyl, 60 cine, ketobemidone, meptazinol, o-desmethyltramadol, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, sub pentazocine, phenazocine, and tapentadol. stituted alkynyl, aryl, Substituted aryl, acyl, Substituted acyl, In certain instances, X is a residue of an amide-containing carboxyl, alkoxycarbonyl, Substituted alkoxycarbonyl, ami opioid, wherein —C(O) N(A ring)-Y-(CRR) noacyl, Substituted aminoacyl, amino, Substituted amino, NH CO) CH(R) N(R) C(O) CH(R) acylamino, Substituted acylamino, and cyano; or 65 N(R) R' is connected to the amide-containing opioid R" and R together with the carbon to which they are through the oxygen of the amide group, wherein the amide attached can form a cycloalkyl or Substituted cycloalkyl group is converted to an amide enol or an imine tautomer. US 8,685,916 B2 53 54 In certain instances, X is an amide-containing opioid, In certain instances, Y is acyl or Substituted acyl. In certain wherein the opioid is selected from alfentanil, carfentanil, instances, Y is acyl. In certain instances, Y is Substituted acyl. fentanyl, lofentanil, loperamide, olmefentanyl, remifentanil, In certain instances, Y is carboxyl. In certain instances, Y is and Sufentanil. alkoxycarbonyl or substituted alkoxycarbonyl. In certain Informulae XIII-XV. A. A. A. and Aare independently instances, Y is alkoxycarbonyl. In certain instances, Y is selected from carbon, nitrogen, oxygen, and Sulfur. In certain Substituted alkoxycarbonyl. In certain instances. Y is aminoa cyl or Substituted aminoacyl. In certain instances, Y is ami instances, A, A, A, and Aare independently selected from noacyl. In certain instances, Y is Substituted aminoacyl. In carbon and nitrogen. In certain instances, A, A, A, and A certain instances, Y is amino or Substituted amino. In certain are independently selected from carbon and oxygen. In cer instances, Y is amino. In certain instances, Y is Substituted tain instances, A', A, A, and Aare independently selected 10 amino. In certain instances, Y is acylamino or Substituted from carbon and sulfur. In certain instances, A, A, A, and acylamino. In certain instances, Y is acylamino. In certain Aare carbon. instances, Y is Substituted acylamino. In certain instances, Y In formulae XIII-XV. A is carbon or nitrogen. In certain is cyano. instances, A is carbon. In certain instances, A is nitrogen. In certain instances, Y is Substituted alkyl. In certain In certain instances, -(CRR), NH C(O)-CH 15 instances, Y is an alkyl group Substituted with a carboxylic (R) N(R)-C(O)-CH(R) N(R) R7 is attached group Such as a carboxylic acid, alkoxycarbonyl or aminoa to A'. In certain instances, —(CRR), NH C(O)-CH cyl. In certain instances, Y is -(CH2)(CH) COON, (R) N(R)-C(O)-CH(R) N(R) R' is attached -(CH2)(C.H.) COOCH, O -(CH2)(CH)— to A. In certain instances, —(CRR). NH CO)—CH COOCHCH, where q is an integer from one to 10. In certain (R) N(R)-C(O)-CH(R) N(R) R' is attached instances, Y is aminoacyl. In certain instances, Y is an alkyl to A. In certain instances, —(CRR). NH CO)—CH group Substituted with an amino group, Substituted amino, or (R) N(R)-C(O)-CH(R) N(R) R' is attached acylamino. to A. In certain instances, Y is aminoacyl or Substituted aminoa Certain Embodiments of Formulae I-XV cyl. In formulae I-XII, the A ring can be a heterocyclic 5 to 25 12-membered ring. In certain instances, Yisaminoacyl comprising phenylene In certain instances, the A ring is a heterocyclic 5 to diamine. In certain instances, Y is 11-membered ring. In certain instances, the A ring is a het erocyclic 5 to 10-membered ring. In certain instances, the A R O ring is a heterocyclic 5 to 9-membered ring. In certain 30 O 21 M N instances, the A ring is a heterocyclic 5 to 8-membered ring. V In certain instances, the A ring is a heterocyclic 5 to 7-mem N N R10: bered ring. In certain instances, the A ring is a heterocyclic 5 or 6-membered ring. In certain instances, the A ring is a R11 heterocyclic 5-membered ring. 35 In certain instances, the A ring is a heterocyclic 6 to wherein each R" is independently selected from hydrogen, 12-membered ring. In certain instances, the A ring is a het alkyl, substituted alkyl, and acyl and R'' is alkyl or substi erocyclic 6 to 11-membered ring. In certain instances, the A tuted alkyl. In certain instances, at least one of R" is acyl. In ring is a heterocyclic 6 to 10-membered ring. In certain certain instances, at least one of R' is alkyl or substituted instances, the A ring is a heterocyclic 6 to 9-membered ring. 40 In certain instances, the A ring is a heterocyclic 6 to 8-mem alkyl. In certain instances, at least one of R" is hydrogen. In bered ring. In certain instances, the A ring is a heterocyclic 6 certain instances, both of R'' are hydrogen. or 7-membered ring. In certain instances, the A ring is a In certain instances, Y is heterocyclic 6-membered ring. In certain instances, the A ring is a heterocyclic 7-membered ring. In certain instances, the A 45 ring is a heterocyclic 8-membered ring. In formulae I-XII, c can be a number from Zero to 3. In certain instances, c is Zero. In certain instances, c is 1. In certain instances, c is 2. In certain instances, c is 3. In formulae I-XV, each Y is independently selected from 50 alkyl, Substituted alkyl, alkenyl, Substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, acyl, substituted wherein R' is hydrogen, alkyl, substituted alkyl, or acyl. In acyl, carboxyl, alkoxycarbonyl, Substituted alkoxycarbonyl, certain instances, R' is acyl. In certain instances, R' is alkyl aminoacyl, Substituted aminoacyl, amino, Substituted amino, or substituted alkyl. In certain instances, R' is hydrogen. acylamino, Substituted acylamino, and cyano. 55 In formulae I-XV. Y can be carboxyl or amino. In certain In certain instances, Y is instances, Y is carboxyl. In certain instances, Y is amino. In certain instances, Y is alkyl or substituted alkyl. In certain instances, Y is alkyl. In certain instances, Y is Substi tuted alkyl. In certain instances, Y is alkenyl or substituted 60 alkenyl. In certain instances, Y is alkenyl. In certain instances, Y is Substituted alkenyl. In certain instances, Y is alkynyl or Sub stituted alkynyl. In certain instances, Y is alkynyl. In certain instances, Y is substituted alkynyl. In certain instances, Y is 65 wherein each R" is independently hydrogen, alkyl, substi aryl or Substituted aryl. In certain instances, Y is aryl. In tuted alkyl, or acyl and b is a number from one to 5. In certain certain instances, Y is substituted aryl. instances, Y is US 8,685,916 B2 55 56 In formulae I-XV, each R" is independently selected from O O hydrogen, alkyl, Substituted alkyl, alkenyl, Substituted alk enyl, alkynyl. Substituted alkynyl, aryl, Substituted aryl, acyl, N Substituted acyl, carboxyl, alkoxycarbonyl, Substituted x-r SR 10. alkoxycarbonyl, aminoacyl, Substituted aminoacyl, amino, O Substituted amino, acylamino, Substituted acylamino, and cyano. wherein each R" is independently hydrogen, alkyl, substi In certain instances, R is hydrogen. In certain instances, tuted alkyl, or acyl. In certain instances, Y is R" is alkyl or substituted alkyl. In certain instances, R is 10 alkyl. In certain instances, R' is substituted alkyl. In certain instances, R' is alkenyl or substituted alkenyl. In certain instances, R' is alkenyl. In certain instances, R' is substituted alkenyl. In certain instances, R' is alkynylor substituted alky nyl. In certain instances, R' is alkynyl. In certain instances, R' 15 is substituted alkynyl. In certain instances, R' is aryl or sub stituted aryl. In certain instances, R' is aryl. In certain R10a O instances, R is substituted aryl. In certain instances, R' is acyl or substituted acyl. In certain instances, R' is acyl. In wherein R" is alkyl and each R" is independently hydro certain instances, R' is substituted acyl. In certain instances, gen, alkyl, Substituted alkyl, or acyl. R" is carboxyl. In certain instances, R' is alkoxycarbonyl or In certain instances, Y is Substituted alkoxycarbonyl. In certain instances, R' is alkoxycarbonyl. In certain instances, R' is substituted alkoxycarbonyl. In certain instances, R' is aminoacyl or Sub O stituted aminoacyl. In certain instances, R' is aminoacyl. In 25 certain instances, R' is substituted aminoacyl. In certain x. N OH: instances, R' is amino or substituted amino. In certain O instances, R' is amino. In certain instances, R' is substituted R O amino. In certain instances, R' is acylamino or substituted acylamino. In certain instances, R' is acylamino. In certain 30 instances, R' is substituted acylamino. In certain instances, wherein R' is independently hydrogen, alkyl, substituted R" is cyano. alkyl, or acyl and b is a number from one to 5. In certain Informulae I-XV, each R is independently selected from instances, Y is hydrogen, alkyl, Substituted alkyl, alkenyl, Substituted alk enyl, alkynyl. Substituted alkynyl, aryl, Substituted aryl, acyl, 35 Substituted acyl, carboxyl, alkoxycarbonyl, Substituted O alkoxycarbonyl, aminoacyl, Substituted aminoacyl, amino, OH: Substituted amino, acylamino, Substituted acylamino, and N cyano. R10 O In certain instances, R is hydrogen. In certain instances, 40 R’ is alkyl or substituted alkyl. In certain instances, R is alkyl. In certain instances, R is substituted alkyl. In certain wherein R' is independently hydrogen, alkyl, substituted instances, R is alkenyl or substituted alkenyl. In certain alkyl, or acyl. instances, R is alkenyl. In certain instances, R is substituted In certain instances, Y is an aminoacyl group. Such as alkenyl. In certain instances, R is alkynylor substituted alky C(O)NR'R'', wherein each R'' and R' is indepen 45 nyl. In certain instances, R is alkynyl. In certain instances, R dently selected from hydrogen, alkyl, Substituted alkyl, and is substituted alkynyl. In certain instances, R is aryl or sub acyl. In certain instances, Y is an aminoacyl group, such as stituted aryl. In certain instances, R is aryl. In certain C(O)NR'R'', wherein R' is an alkyl and R' is sub instances, R is substituted aryl. In certain instances, R is stituted alkyl. In certain instances, Y is an aminoacyl group, acyl or substituted acyl. In certain instances, R is acyl. In 50 certain instances, R is substituted acyl. In certain instances, such as –C(O)NR'R'' is wherein R' is an alkyl and R’ is carboxyl. In certain instances, R is alkoxycarbonyl or R' is alkyl substituted with a carboxylic acid oralkoxycar substituted alkoxycarbonyl. In certain instances, R is bonyl. In certain instances, Y is an aminoacyl group. Such as alkoxycarbonyl. In certain instances, R is substituted C(O)NR'R'', wherein R' is methyl and R' is alkyl alkoxycarbonyl. In certain instances, R is aminoacyl or Sub substituted with a carboxylic acid or alkoxycarbonyl. 55 stituted aminoacyl. In certain instances, R is aminoacyl. In In certain instances, Y is carboxyl. certain instances, R is substituted aminoacyl. In certain In certain instances, Y is acyl or Substituted acyl. instances, R is amino or substituted amino. In certain In certain instances, Y is alkoxycarbonyl or substituted instances, R is amino. In certain instances, R is substituted alkoxycarbonyl. amino. In certain instances, R is acylamino or substituted In certain instances, Y is amino or Substituted amino. 60 acylamino. In certain instances, R is acylamino. In certain In certain instances, Y is acylamino or Substituted acy instances, R is substituted acylamino. In certain instances, lamino. R’ is cyano. In formulae I-XV, a can be an integer from one to 8. In In certain instances, one of R' and R is hydrogen. In certain instances, a is one. In certain instances, a is 2. In certain instances, one of R' and R is alkyl. In certain certain instances, a is 3. In certain instances, a is 4. In certain 65 instances, one of R' and R is substituted alkyl. In certain instances, a is 5. In certain instances, a is 6. In certain instances, one of R' and R is alkenyl or substituted alkenyl. instances, a is 7. In certain instances, a is 8. In certain instances, one of RandR is alkynyl or substituted US 8,685,916 B2 57 alkynyl. In certain instances, one of R' and R is aryl or substituted aryl. In certain instances, one of R' and R is acyl or substituted acyl. In certain instances, one of R' and R is carboxyl. In certain instances, one of R' and R is alkoxycar bonyl or Substituted alkoxycarbonyl. In certain instances, one of RandR is aminoacylor substitutedaminoacyl. In certain instances, one of R' and R is amino or substituted amino. In certain instances, one of RandR is acylamino or substituted acylamino. In certain instances, one of R' and R is cyano. wherein R' is hydrogen, alkyl, substituted alkyl, or acyl. In In certain instances, R' and R are hydrogen. In certain 10 certain instances, R' is acyl. In certain instances, R' is alkyl instances, R' and R on the same carbon are both alkyl. In or substituted alkyl. In certain instances, R' is hydrogen. certain instances, RandR on the same carbon are methyl. In In certain instances, one of R and R is certain instances, R' and R on the same carbon are ethyl. In certain instances, R' and R' which are vicinal are both alkyl and RandR which are vicinal are both hydrogen. In 15 O R10 certain instances, R' and R' which are vicinal are both ethyl and RandR which are vicinal are both hydrogen. In certain N instances, RandR' which are vicinal are both methylandR X's.O n R10; and R which are vicinal are both hydrogen. R O In certain instances, in the chain of -IC(R')(R)). , not every carbon is Substituted. In certain instances, in the chain of C(R')(R)). , there is a combination of different alkyl wherein each R' is independently hydrogen, alkyl, substi Substituents, such as methyl or ethyl. tuted alkyl, or acyl and b is a number from one to 5. In certain In certain instances, one or both of RandR is substituted instances, one of R' and R is alkyl. In certain instances, one or both of RandR is an alkyl 25 group Substituted with a carboxylic group Such as a carboxy lic acid, alkoxycarbonyl or aminoacyl. In certain instances, O R10 one or both of R and R is -(CH2)(C.H.) COOH, N -(CH2)(CH) COOCHs. O -(CH2)(CH)— COOCHCH, where q is an integer from one to 10. In certain 30 instances, one or both of R and R is aminoacyl. R10 O In formulae I-XV. R' and R together with the carbon to which they are attached can form a cycloalkyl or substituted wherein each R" is independently hydrogen, alkyl, substi cycloalkyl group, or two R' or R groups on adjacent carbon tuted alkyl, or acyl. In certain instances, one of R' and R is atoms, together with the carbon atoms to which they are 35 attached, can form a cycloalkyl or Substituted cycloalkyl group. In certain instances, R' and R together with the car O R10 bon to which they are attached can form a cycloalkyl group. Thus, in certain instances, RandR on the same carbon form N a spirocycle. In certain instances, RandR together with the 40 carbon to which they are attached can form a substituted k O cycloalkyl group. In certain instances, two R' or R groups on adjacent carbon atoms, together with the carbon atoms to wherein R' is alkyl and each R' is independently hydro which they are attached, can form a cycloalkyl group. In gen, alkyl, Substituted alkyl, or acyl. certain instances, two R' or R groups on adjacent carbon 45 In certain instances, one of R' and R is atoms, together with the carbon atoms to which they are attached, can form a Substituted cycloalkyl group. In certain instances, one of R' and R is aminoacyl. O In certain instances, one of R and R is aminoacyl com prising phenylenediamine. In certain instances, one or both of 50 R" and R is x OH: O R O R10 O 4N / 55 wherein R' is independently hydrogen, alkyl, substituted --N alkyl, or acyl and b is a number from one to 5. In certain 1s Yi. instances, one of R' and R is R11 60 O wherein each R' is independently selected from hydrogen, OH: alkyl, substituted alkyl, and acyl and R'' is alkyl or substi N tuted alkyl. In certain instances, at least one of R' is acyl. In certain instances, at least one of R' is alkyl or substituted R10 O alkyl. In certain instances, at least one of R" is hydrogen. In 65 certain instances, both of R'' are hydrogen. wherein R' is independently hydrogen, alkyl, substituted In certain instances, one of R' and R is alkyl, or acyl. US 8,685,916 B2 59 60 In certain instances, one of R' and R is an aminoacyl In certain instances, when a is one, the A ring is a hetero group, such as –C(O)NR'R'', wherein each R'' and cyclic 6 to 11-membered ring. In certain instances, whena is R" is independently selected from hydrogen, alkyl, substi one, the A ring is a heterocyclic 6 to 10-membered ring. In tuted alkyl, and acyl. In certain instances, one of RandR is certain instances, whena is one, the A ring is a heterocyclic 6 an aminoacyl group, such as C(O)NR'R'', wherein to 9-membered ring. In certain instances, whena is one, the A R" is an alkyl and R' is substituted alkyl. In certain ring is a heterocyclic 6 to 8-membered ring. In certain instances, one of R' and R is an aminoacyl group, such as instances, when a is one, the A ring is a heterocyclic 6 to C(O)NR'R'', wherein R' is an alkyl and R' is alkyl 7-membered ring. In certain instances, when a is one, the A substituted with a carboxylic acid or alkoxycarbonyl. In cer ring is a heterocyclic 6-membered ring. tain instances, one of R' and R is an aminoacyl group, such 10 as C(O)NR'R'', wherein R' is methyl and R' is In certain instances, when the A ring is a heterocyclic alkyl substituted with a carboxylic acid or alkoxycarbonyl. 5-membered ring, then a is an integer from 2 to 7. In certain In certain instances, one of R' and Rare carboxyl. instances, when the A ring is a heterocyclic 5-membered ring, In certain instances, one of R' and R is acyl or substituted thena is an integer from 2 to 6. In certain instances, when the acyl. 15 A ring is a heterocyclic 5-membered ring, thena is an integer In certain instances, one of R' and R is alkoxycarbonyl or from 2 to 5. In certain instances, when the A ring is a hetero substituted alkoxycarbonyl. cyclic 5-membered ring, then a is an integer from 2 to 4. In In certain instances, one of R' and R is amino or substi certain instances, when the A ring is a heterocyclic 5-mem tuted amino. bered ring, then a is an integer from 2 to 3. In certain In certain instances, one of R' and R is acylamino or instances, when the A ring is a heterocyclic 5-membered ring, Substituted acylamino. thena is 2. In certain instances, R' or R can modulate a rate of In certain instances, the A ring is a heterocyclic 7-mem intramolecular cyclization. R' or R can speed up a rate of bered or 8-membered ring and a is 1 or 2. In certain instances, intramolecular cyclization, when compared to the corre the A ring is a heterocyclic 7-membered ring and a is 1. In sponding molecule where R' and R are both hydrogen. In 25 certain instances, the A ring is a heterocyclic 7-membered certain instances, R' or R comprise an electron-withdrawing ringanda is 2. In certain instances, the A ring is a heterocyclic group or an electron-donating group. In certain instances, R' 8-membered ring and a is 1. In certain instances, the A ring is or R comprise an electron-withdrawing group. In certain a heterocyclic 8-membered ring and a is 2. instances, R' or R comprise an electron-donating group. In certain instances, a certain group of compounds are Atoms and groups capable of functioning as electron-with 30 compounds of formulae I-XII, wherein A ring is a 5-mem drawing Substituents are well known in the field of organic bered ring and a is 2. A certain group of compounds are chemistry. They include electronegative atoms and groups compounds of formulae I-XII, wherein A ring is a 6-mem containing electronegative atoms. Such groups function to bered ring and a is one. lower the basicity or protonation state of a nucleophilic nitro Informulae I-XV, each R can independently be hydrogen, gen in the beta position via inductive withdrawal of electron 35 alkyl, substituted alkyl, aryl or substituted aryl. density. Such groups can also be positioned on other positions In certain instances, at least one R is hydrogen. In certain along the alkylene chain. Examples include halogen atoms instances, at least one R is alkyl. In certain instances, at least (for example, a fluorine atom), acyl groups (for example an one R is substituted alkyl. In certain instances, at least one R alkanoyl group, an aroyl group, a carboxyl group, an alkoxy is aryl. In certain instances, at least one R is substituted aryl. carbonyl group, an aryloxycarbonyl group or an aminocarbo 40 In certain instances, each of the R is hydrogen or alkyl. In nyl group (Such as a carbamoyl, alkylaminocarbonyl, dialky certain instances, all Rare hydrogen. In certain instances, all laminocarbonyl or arylaminocarbonyl group)), an oxo (=O) R are alkyl. In certain instances, the R of N R that is Substituent, a nitrile group, a nitro group, ether groups (for adjacent to C R is hydrogen or alkyl. In certain instances, example an alkoxy group) and phenyl groups bearing a Sub the R of N-R that is adjacent to C R is hydrogen. In stituent at the ortho position, the para position or both the 45 certain instances, the RofN R that is adjacent to C R is ortho and the para positions, each Substituent being selected alkyl. independently from a halogenatom, a fluoroalkyl group (Such In formulae I-III, VI-VII, X, and XIII, Rican be selected as trifluoromethyl), a nitro group, a cyano group and a car from hydrogen, alkyl, Substituted alkyl, aryl, Substituted aryl, boxyl group. Each of the electron withdrawing substituents arylalkyl, substituted arylalkyl, heteroalkyl, substituted het can be selected independently from these. 50 eroalkyl, heteroaryl, substituted heteroaryl, heteroarylalkyl, In certain instances, -IC(R')(R) is selected from and substituted heteroarylalkyl. In certain instances, informulae I-III, VI-VII, X, and XIII, R is selected from hydrogen, alkyl, substituted alkyl, aryl, substituted aryl, arylalkyl, substituted arylalkyl, heteroalkyl, 55 substituted heteroalkyl, heteroaryl, substituted heteroaryl, CHCH(C(O)OR) : CHCH(C(O)OH)–: -CH heteroarylalkyl, and substituted heteroarylalkyl. In certain (CHF)CHCH(CHF) : CH(CHF)CHCH(CHF) : instances, R is selected from hydrogen, alkyl, substituted —CH(CF)CH-CH(CF) : - CHCH-CH(CF) : alkyl, arylalkyl, substituted arylalkyl, heteroarylalkyl, and —CHCH-CH(CHF)–: - CHCH-CH(CHF) : substituted heteroarylalkyl. In certain instances, R is hydro CHCH-CH(C(O)NR'R') : CHCH-CH(C(O) 60 gen. In certain instances, R is alkyl. In certain instances, R OR)-; and CHCH-CH(C(O)OH) , in which R', is substituted alkyl. In certain instances, R is arylalkyl or R", R’ and Reach independently represents hydrogen or substituted arylalkyl. In certain instances, R is heteroaryla (1-6C)alkyl, and R and Reach independently represents lkyl or substituted heteroarylalkyl. (1-6C)alkyl. In certain instances, informulae I-III, VI-VII, X, and XIII, Informulae I-XII, whena is one, the Aring is a heterocyclic 65 R is a side chain of an amino acid, a side chain of an amino 6 to 12-membered ring; and when the A ring is a heterocyclic acid variant, a derivative of a side chain of an amino acid, or 5-membered ring, then a is an integer from 2 to 8. a derivative of a side chain of an amino acid variant. US 8,685,916 B2 61 62 Informulae IV, VIII, XI, and XIV, Rican be a side chain of cleavable moiety. A GI enzyme-cleavable moiety is a struc an amino acid selected from alanine, arginine, asparagine, tural moiety that is capable of being cleaved by a GI enzyme. aspartic acid, cysteine, glutamic acid, glutamine, glycine, A trypsin-cleavable moiety is a structural moiety that is histidine, isoleucine, leucine, lysine, methionine, phenylala capable of being cleaved by trypsin. In certain instances, a GI nine, proline, serine, threonine, tryptophan, tyrosine, Valine, enzyme-cleavable moiety comprises a charged moiety that homoarginine, homolysine, ornithine, arginine mimic, argi can fit into an active site of a GI enzyme and is able to orient nine homologue, arginine truncate, arginine with varying oxi the prodrug for cleavage at a Scissile bond. In certain dation states, lysine mimic, lysine homologue, lysine trun instances, a trypsin-cleavable moiety comprises a charged cate, and lysine with varying oxidation states. moiety that can fit into an active site of trypsin and is able to In certain instances, Rican be a side chain of an amino acid 10 selected from alanine, arginine, asparagine, aspartic acid, orient the prodrug for cleavage at a scissile bond. For cysteine, glutamic acid, glutamine, glycine, histidine, isoleu instance, the charged moiety of a GI enzyme-cleavable moi cine, leucine, lysine, methionine, phenylalanine, proline, ety. Such as a trypsin-cleavable moiety, can be a basic moiety serine, threonine, tryptophan, tyrosine, and valine. that exists as a charged moiety at physiological pH. A deriva tive of an amino acid or of an amino acid variant refers to a In certain instances, R is a side chain of an L-amino acid 15 selected from L-alanine, L-arginine, L-asparagine, L-aspartic substance that has been altered from another substance by acid, L-cysteine, L-glutamic acid, L-glutamine, glycine, modification, partial Substitution, homologation, truncation, L-histidine, L-isoleucine, L-leucine, L-lysine, L-methionine, or a change in oxidation state, while retaining the ability to be L-phenylalanine, L-proline, L-serine, L-threonine, L-tryp cleaved by a GI enzyme. tophan, L-tyrosine, L-Valine, L-homoarginine, L-homol For example, to form a trypsin-cleavable moiety, R can ysine, L-ornithine, L-arginine mimic, L-arginine homologue, include, but is not limited to, a side chain of lysine (such as L-arginine truncate, L-arginine with varying oxidation states, L-lysine), arginine (Such as L-arginine), homolysine, L-lysine mimic, L-lysine homologue, L-lysine truncate, and homoarginine, and ornithine. Other values for Rinclude, but L-lysine with varying oxidation states. are not limited to, a side chain of an arginine mimic, arginine In certain instances, R is a side chain of an L-amino acid 25 homologue, arginine truncate, arginine with varying oxida selected from L-alanine, L-arginine, L-asparagine, L-aspartic tion states (for instance, metabolites), lysine mimic, lysine acid, L-cysteine, L-glutamic acid, L-glutamine, glycine, homologue, lysine truncate, and lysine with varying oxida L-histidine, L-isoleucine, L-leucine, L-lysine, L-methionine, tion states (for instance, metabolites). Examples of arginine L-phenylalanine, L-proline, L-serine, L-threonine, L-tryp and lysine mimics include arylguanidines, arylamidines (Sub tophan, L-tyrosine, and L-valine. 30 stituted benzamidines), benzylamines, (bicyclo[2.2.2]octan In certain instances, R is a side chain of an amino acid 1-yl)methanamine, citrulline, homocitrulline and derivatives selected from arginine, lysine, homoarginine, homolysine, thereof. ornithine, arginine mimic, arginine homologue, arginine In certain instances, R is a side chain of an amino acid that truncate, arginine with varying oxidation states, lysine effects—C(O)—C(R) N(R)—to be a GI enzyme-cleav mimic, lysine homologue, lysine truncate, and lysine with 35 able moiety, such as a trypsin-cleavable moiety. In certain varying oxidation states. Examples of arginine and lysine instances, R is a side chain of an amino acid variant that mimics include arylguanidines, arylamidines (substituted effects—C(O)—C(R) N(R)—to be a GI enzyme-cleav benzamidines), benzylamines and (bicyclo2.2.2]octan-1-yl) able moiety, such as a trypsin-cleavable moiety. In certain methanamine, citrulline, homocitrulline and derivatives instances, R is a derivative of a side chain of an amino acid thereof. In certain instances, R is a side chain of an amino 40 that effects - C(O) C(R) N(R)- to be a GI enzyme acid selected from arginine, lysine, homoarginine, homol cleavable moiety, such as a trypsin-cleavable moiety. In cer ysine, and ornithine. In certain instances, R is a side chain of tain instances, R is a derivative of a side chain of an amino an amino acid selected from arginine or lysine. In certain acid variant that effects—C(O) C(R) N(R)—to be a GI instances, R is a side chain of arginine. In certain instances, enzyme-cleavable moiety, such as a trypsin-cleavable moiety. R is a side chain of lysine. 45 In certain instances, R is a side chain of an amino acid In certain instances, R is a side chain of an L-amino acid selected from arginine, lysine, homoarginine, homolysine, selected from L-arginine, L-lysine, L-homoarginine, L-ho ornithine, arginine mimic, arginine homologue, arginine molysine, L-ornithine, L-arginine mimic, L-arginine homo truncate, arginine with varying oxidation states, lysine logue, L-arginine truncate, L-arginine with varying oxidation mimic, lysine homologue, lysine truncate, and lysine with states, L-lysine mimic, L-lysine homologue, L-lysine trun 50 varying oxidation states. In certain instances, R is a side cate, and L-lysine with varying oxidation states. In certain chain of an amino acid selected from arginine, lysine, instances, R is a side chain of an L-amino acid selected from homoarginine, homolysine, and ornithine. In certain L-arginine, L-lysine, L-homoarginine, L-homolysine, and instances, R is a side chain of an amino acid selected from L-ornithine. In certain instances, R is a side chain of an arginine or lysine. In certain instances, R is a side chain of L-amino acid selected from L-arginine or L-lysine. In certain 55 arginine. In certain instances, R is a side chain of lysine. instances, R is a side chain of L-arginine. In certain In certain instances, R is a side chain of an L-amino acid instances, R is a side chain of L-lysine. selected from L-arginine, L-lysine, L-homoarginine, L-ho In certain instances, R molysine, L-ornithine, L-arginine mimic, L-arginine homo represents CHCHCH-NH(C(=NH)(NH)) O logue, L-arginine truncate, L-arginine with varying oxidation —CH2CH2CHCH-NH2, the configuration of the carbon 60 states, L-lysine mimic, L-lysine homologue, L-lysine trun atom to which R is attached corresponding with that in an cate, and L-lysine with varying oxidation states. In certain L-amino acid. instances, R is a side chain of an L-amino acid selected from Informulae V, IX, XII, and XV. Rican be a side chain of an L-arginine, L-lysine, L-homoarginine, L-homolysine, and amino acid, a side chain of an amino acid variant, a derivative L-ornithine. In certain instances, R is a side chain of an of a side chain of an amino acid, or a derivative of a side chain 65 L-amino acid selected from L-arginine and L-lysine. In cer of an amino acid variant that effects —C(O) C(R) N tain instances, R is a side chain of L-arginine. In certain (R)—to be a GIenzyme-cleavable moiety, such as a trypsin instances, US 8,685,916 B2 63 64 R is a side chain of L-lysine. to R is a side chain of L-alanine. In certain instances, R that In certain instances, R is immediately adjacent to R is a side chain of glycine. represents CHCHCH-NH(C(=NH)(NH)) O Informulae I-XV. R' can be selected from hydrogen, alkyl, —CH2CH2CHCH-NH2, the configuration of the carbon Substituted alkyl, acyl, Substituted acyl, alkoxycarbonyl, Sub atom to which R is attached corresponding with that in an 5 stituted alkoxycarbonyl, aryl, Substituted aryl, arylalkyl, and L-amino acid. substituted arylalkyl. Informulae I-XV, b is a number from Zero to 100. In certain In certain instances, R is hydrogen, alkyl, acyl, or substi instances, b is Zero to 50. In certain instances, b is zero to 90, tuted acyl. In certain instances, R is hydrogen, acyl, or sub 80, 70, 60, 50, 40, 30, 20, or 10. In certain instances, b is 100. stituted acyl. In certain instances, R is hydrogen. In certain 10 instances, R is alkyl. In certain instances, R is acyl or sub In certain instances, b is 75. In certain instances, b is 50. In stituted acyl. In certain instances, R is acyl. In certain certain instances, b is 25. In certain instances, b is 20. In instances, R is substituted acyl. In certain instances, R' can certain instances, b is 15. In certain instances, b is 10. In be acetyl, benzoyl, malonyl, piperonyl or Succinyl. In certain certain instances, b is 9. In certain instances, b is 8. In certain instances, R' can be acetyl. In certain instances, R' can be instances, b is 7. In certain instances, b is 6. In certain 15 malonyl. instances, b is 5. In certain instances, b is 4. In certain In certain instances, a certain group of compounds are instances, b is 3. In certain instances, b is 2. In certain compounds of formulae I-XV, wherein R is a side chain of an instances, b is one. In certain instances, b is Zero. In certain amino acid selected from arginine and lysine and b is one. In instances, b is Zero or one. In certain instances, b is Zero or one certain instances, a certain group of compounds are com Or tWO. 2O pounds of formulae I-XV, wherein R is a side chain of an In formulae I-III, VI-VII, X, and XIII, each R can be amino acid selected from arginine and lysine; R is a side independently selected from hydrogen, alkyl, Substituted chain of an amino acid selected from alanine and glycine; and alkyl, aryl, substituted aryl, arylalkyl, substituted arylalkyl, b is one. In certain instances, a certain group of compounds heteroalkyl, substituted heteroalkyl, heteroaryl, substituted are compounds of formulae I-XV, wherein R is a side chain heteroaryl, heteroarylalkyl, and substituted heteroarylalkyl. 25 of an amino acid selected from L-arginine and L-lysine and b In certain instances, formulae I-III, VI-VII, X, and XIII, is one. In certain instances, a certain group of compounds are each R is independently selected from hydrogen, alkyl, sub compounds of formulae I-XV, wherein R is a side chain of an stituted alkyl, aryl, substituted aryl, arylalkyl, substituted ary amino acid selected from L-arginine and L-lysine; R is a side lalkyl, heteroalkyl, substituted heteroalkyl, heteroaryl, sub chain of an amino acid selected from L-alanine and glycine; stituted heteroaryl, heteroarylalkyl, and substituted 30 and b is one. In certain instances, a certain group of com heteroarylalkyl. In certain instances, R is hydrogen. In cer pounds are compounds of formulae I-XV, wherein Risa side tain instances, R is alkyl. In certain instances, R is substi chain of L-arginine; R is a side chain of an amino acid tuted alkyl. In certain instances, R is arylalkyl or substituted selected from L-alanine and glycine; and b is one. In certain arylalkyl. In certain instances, R is heteroarylalkyl or substi instances, a certain group of compounds are compounds of tuted heteroarylalkyl. 35 formulae I-XV, wherein R is a side chain of L-lysine; R is a In certain instances, formulae I-III, VI-VII, X, and XIII, R side chain of an amino acid selected from L-alanine and is a side chain of an amino acid, a side chain of an amino acid glycine; and b is one. variant, a derivative of a side chain of an amino acid, or a In certain instances, a certain group of compounds are derivative of a side chain of an amino acid variant. In certain compounds of formulae I-XV, wherein R is a side chain of instances, R is a side chain of an amino acid. In certain 40 glycine and b is one. In certain instances, a certain group of instances, R is a side chain of an amino acid variant. In compounds are compounds of formulae I-XV, wherein R is certain instances, Risa derivative of a side chain of an amino a side chain of an amino acid selected from arginine and acid. In certain instances, R is a derivative of a side chain of lysine; R is a side chain of glycine; and b is one. In certain an amino acid variant. instances, a certain group of compounds are compounds of Informulae IV, V, VIII, IX, XI, XII, XIV, and XV each R 45 formulae I-XV, wherein R is a side chain of an amino acid is a side chain of an amino acid independently selected from selected from L-arginine and L-lysine; R is a side chain of alanine, arginine, asparagine, aspartic acid, cysteine, glycine; and b is one. glutamic acid, glutamine, glycine, histidine, isoleucine, leu In certain instances, a certain group of compounds are cine, lysine, methionine, phenylalanine, proline, serine, compounds of formulae I-XV, wherein R is a side chain of threonine, tryptophan, tyrosine and valine. 50 alanine and b is one. In certain instances, a certain group of In certain instances, R is a side chain of an L-amino acid compounds are compounds of formulae I-XV, wherein R is selected from L-alanine, L-arginine, L-asparagine, L-aspartic a side chain of L-alanine and b is one. In certain instances, a acid, L-cysteine, L-glutamic acid, L-glutamine, glycine, his certain group of compounds are compounds of formulae tidine, L-isoleucine, L-leucine, L-lysine, L-methionine, I-XV, wherein R is a side chain of an amino acid selected L-phenylalanine, L-proline, L-serine, L-threonine, L-tryp- 55 from arginine and lysine; R is a side chain of alanine; and b tophan, L-tyrosine, and L-valine. is one. In certain instances, a certain group of compounds are In certain instances, R that is immediately adjacent to R compounds of formulae I-XV, wherein R is a side chain of an represents —H or —CH, the configuration of the carbon amino acid selected from L-arginine and L-lysine; R is a side atom to which R is attached corresponding with that in an chain of L-alanine; and b is one. L-amino acid. In certain instances, R that is immediately 60 In certain instances, a certain group of compounds are adjacent to R represents —H. In certain instances, R that is compounds of formulae I-XV, wherein R is a side chain of immediately adjacent to R represents —CHs, the configura L-arginine: R is a side chain of L-alanine; and b is one. In tion of the carbonatom to which R is attached corresponding certain instances, a certain group of compounds are com with that in an L-amino acid. pounds of formulae I-XV, wherein R is a side chain of L-argi In certain instances, R that is immediately adjacent to R 65 nine; R is a side chain of glycine; and b is one. In certain is a side chain of an amino acid selected from L-alanine and instances, a certain group of compounds are compounds of glycine. In certain instances, R that is immediately adjacent formulae I-XV, wherein R is a side chain of L-lysine; R is a US 8,685,916 B2 65 66 side chain of L-alanine; and b is one. In certain instances, a Certain examples of amino acid variants include, but are certain group of compounds are compounds of formulae not limited to: dehydroalanine, ethionine, hypusine, lanthion I-XV, wherein R is a side chain of L-lysine; R is a side chain ine, pyrrolysine, C.-aminoisobutyric acid, selenomethionine of glycine; and b is one. Amino Acids Found in Prodrugs and derivatives thereof. Amino acid means a building block of a polypeptide. As Certain examples of amino acid variants include, but are used herein, “amino acid includes the 20 common naturally not limited to: (3.2-amino benzoic acid, 2-amino methylben occurring L-amino acids and all amino acids variants. In Zoic acid, 2-amino-3-guanidinopropionic acid, 2-amino-3- certain embodiments, an amino acid is a cleavable Substrate methoxy benzoic acid, 2-amino-3-ureidopropionic acid, for a gastrointestinal enzyme. 10 3-amino benzoic acid, 4-amino benzoic acid, 4-amino methyl “Naturally occurring amino acids' means the 20 common benzoic acid, 4-nitroanthranillic acid, 5-acetamido-2-ami naturally occurring L-amino acids, that is, alanine, arginine, nobenzoic acid, butanoic acid (HMB), glutathione, homocys asparagine, aspartic acid, cysteine, glutamic acid, glutamine, teine, statine, taurine, B-alanine, 2-hydroxy-4-(methylthio), glycine, histidine, isoleucine, leucine, lysine, methionine, 15 (3,4)-diamino benzoic acid, (3,5)-diamino benzoic acid and phenylalanine, proline, serine, threonine, tryptophan, derivatives thereof. tyrosine and valine. Certain examples of amino acid variants include, but are Amino acid variants' means an amino acid other than any not limited to: (2 amino ethyl). cysteine, 2-amino-3-ethyOX of the 20 common naturally occurring L-amino acids that is ybutanoic acid, buthionine, cystathion, cysteic acid, ethion hydrolyzable by a protease in a manner similar to the ability ine, ethoxytheorine, methylserine, N-e-e-dimethyl-lysin, of a protease to hydrolyze a naturally occurring L-amino acid. N-(p-nitro-arginine, Saccharopine, isoserine derivatives Amino acid variants, thus, include amino acids or analogs of thereof, and combinations thereof. amino acids other than the 20 naturally-occurring amino Certain examples of amino acid variants include, but are acids. Amino acid variants include synthetic amino acids. 25 The embodiments also include derivatives of amino acids not limited to: 1-carnitine, selenocysteine, 1-sarcosine, l-lysi and of amino acid variants. A derivative of anamino acid or of nol, benzoic acid, citric acid, choline, EDTA or Succinic acid an amino acid variant refers to a Substance that has been and derivatives thereof. altered from another substance by modification, partial sub Certain examples of amino acid variants are amino alco stitution, homologation, truncation, or a change in oxidation 30 hols. Examples of amino alcohols include, but are not limited state, while retaining the ability to be cleaved by a GI enzyme. to: alaminol, indano, norephedrine, asparaginol, aspartimol, Certain examples of amino acid variants include, but are glutamol, leucinol, methioninol, phenylalaminol, prolinol, not limited to: 2-aminoindane-2-carboxylic acid, 2-ami tryptophanol, Valinol, isoleucinol, argininol, serinol, tyrosi noisobutyric acid, 4-amino-phenylalanine, 5-hydroxylysine, 35 nol, threoninol, cysteinol, lysinol, histidinol and derivatives biphenylalanine, citrulline, cyclohexylalanine, cyclohexylg thereof. lycine, diethylglycine, dipropylglycine, homoarginine, General Synthetic Procedures for Formulae I-XV homocitrulline, homophenylalanine, homoproline, Representative synthetic Schemes for compounds dis homoserine, homotyrosine, hydroxyproline, lanthionine, naphthylalanine, norleucine, ornithine, phenylalanine(4- 40 closed herein are shown below. Compounds of Formulae fluoro), phenylalanine(4-nitro), phenylglycine, pipecolic I-XV can be synthesized by using the disclosed methods. acid, tert-butylalanine, tert-butylglycine, tert-leucine, tet Representative Synthetic Schemes rahydroisoquinoline-3-carboxylic acid, C.-aminobutyric acid, A representative synthesis for Compound S-104 is shown y-aminobutyric acid, 2,3-diaminoproprionic acid, phenyla 45 in Scheme 1. In Scheme 1, R. A ring, Y, and c are defined lanine(2,3,4,5,6 pentafluoro), aminohexanoic acid and herein. PG' is an amino protecting group. Although the derivatives thereof. schemes herein show a morphinan structure for X in Formu Certain examples of amino acid variants include, but are lae I-XV, the entire scope of X as an opioid as applicable to not limited to, N-methyl amino acids. For example, N-me Formula I-XV is contemplated. Also, although the schemes thyl-alanine, N-methyl aspartic acid, N-methyl-glutamic 50 herein show RandR as being hydrogen and a being one, the acid, N-methyl-glycine (sarcosine) are N-methyl amino entire scope of R', R, anda as applicable to Formula I-XV is acids. contemplated.

Scheme 1 -CH3 N Ra ON O O S-101

HC-O S-100 US 8,685,916 B2

-continued CH N1 HN N NH-PG Ra A (Y),

O -e-

HC-O O O usO S-102 N1 CH 3 Ra

HC-O O O ul N YNH-PG A (Y),

S-104

25 In Scheme 1, Compound S-100 is a commercially available formates. In Scheme 1, the activation agent Compound S-101 starting material. Alternatively, Compound S-100 can be is 4-nitrophenyl chloroformate. Other suitable activation semi-synthetically derived from natural materials or synthe agents can be used prior to reaction with Compound S-103. sized via a variety of different synthetic routes using com With continued reference to Scheme 1, Compound S-102 mercially available starting materials and/or starting materi 30 reacts with Compound S-103 to form Compound S-104. In als prepared by conventional synthetic methods. Scheme 1, Compound S-103 is a commercially available With continued reference to Scheme 1, Compound S-100 starting material. Alternatively, Compound S-103 can be syn is enolized. Enolization of a ketone can be performed with thesized via a variety of different synthetic routes using com reaction with a strong base, such as potassium hexamethyld mercially available starting materials and/or starting materi isilazide (KHMDS). The enolate of Compound S-100 is then 35 als prepared by conventional synthetic methods. reacted with an activation agent, such as Compound S-101, to A representative synthesis for Compound S-203 is shown form intermediate Compound S-102. Suitable activation in Scheme 2. In Scheme 2, R". A ring, Y, c, and Rare defined agents include carbonate-forming reagents, such as chloro herein. PG' and PG are amino protecting groups. Scheme 2 N1 CH3 Ra

HC-O O ls N YNH-PG A (Y),

S-104 N1 CH3 Ra O N-PG? O HO R5 O l N N NH2 HC-O A S-2O2

(Y), US 8,685,916 B2 70

entinued3 N1 Ra

HC-O O O ul N NN A H

(Y),

S-2O3

In Scheme 2, the protecting group PG is removed from diisopropylethylamine (DIEA). Suitable coupling reagents Compound S-104 to form Compound S-201. Conditions to for use include, by way of example, carbodiimides, such as remove amino groups can be found in Greene and Wuts. ethyl-3-(3-dimethylamino)propylcarbodiimide (EDC), dicy When PG' is a Boc group, the protecting group can be clohexylcarbodiimide (DCC), diisopropylcarbodiimide (DIC) and the like, and other well-known coupling reagents, removed with acidic conditions. Such as treatment with such as N,N'-carbonyldiimidazole, 2-ethoxy-1-ethoxycarbo hydrochloric acid or trifluoroacetic acid. nyl-1,2-dihydroquinoline (EEDQ), benzotriazol-1-yloxy-tris With reference to Scheme 2, Compound S-201 reacts with (dimethylamino)phosphonium hexafluorophosphate (BOP), Compound S-202 to form Compound S-203 in a peptide O-(7-azabenzotriazol-1-yl)-N.N.N.N',N'-tetramethyluro coupling reaction. In certain embodiments, R is a side chain 25 nium hexafluorophosphate (HATU) and the like. Optionally, of an amino acid and is optionally protected. Protecting well-known coupling promoters, such as N-hydroxysuccin groups for the side chain of amino acids are known to those imide, 1-hydroxybenzotriazole (HOBT), 1-hydroxy-7-aza skilled in art and can be found in Greene and Wuts. In certain benzotriazole (HOAT), N,N-dimethylaminopyridine instances, the protecting group for the side chain of arginine (DMAP) and the like, can be employed in this reaction. Typi is a Sulfonyl-type protecting group, Such as 2.2,4,6,7-pentam 30 cally, this coupling reaction is conducted at a temperature ethyldihydrobenzofurane (Pbf). Other protecting groups ranging from about 0°C. to about 60°C. for about 1 to about include 2,2,5,7,8-pentamethylchroman (Pmc) and 1,2-dim 72 hours in an inert diluent, such as THF or DMF. In certain ethylindole-3-sulfonyl (MIS). instances, Compound S-201 reacts with Compound S-202 to A peptide coupling reaction typically employs a conven form Compound S-203 in the presence of HATU. tional peptide coupling reagent and is conducted under con 35 A representative synthesis for Compound S-303 is shown ventional coupling reaction conditions, typically in the pres in Scheme 3. In Scheme 3, R. A ring, Y, c, R. R. and Rare ence of a trialkylamine. Such as triethylamine or defined herein. PG is an amino protecting group.

Scheme 3 CH3

Ra

O O ls N-PG2 -> HC-O N NN A H R5 (Y),

S-2O3 CH3 O Ra HN-R7 HO

O O R6 l NH, - S-302 - HC-O N NN A H R5 (Y),

S-301 US 8,685,916 B2 71 72 CH -continued N1 3 Ra

HC-O - NN A

In Scheme 3, the protecting group PG is removed from Additional amino acids can be added to the compound Compound S-203 to form Compound S-301. Conditions to through standard peptide coupling reactions as discussed remove amino groups can be found in Greene and Wuts. herein. Removal of other protecting groups can be performed When PG is a Boc group, the protecting group can be if other protecting groups were used, such as protecting removed with acidic conditions. Such as treatment with groups present on the R or R moiety. Conditions for removal hydrochloric acid or trifluoroacetic acid. of other protecting groups depend on the identity of the pro With reference to Scheme 3, Compound S-301 reacts with tecting group and are known to those skilled in the art. The Compound S-302 to form Compound S-303 in a peptide 25 conditions can also be found in Greene and Wuts. coupling reaction. A peptide coupling reaction typically Additional Representative Synthetic Schemes employs a conventional peptide coupling reagent and is con Representative synthesis for Compound S-404 is shown in ducted under conventional coupling reaction conditions, typi Scheme 4. In Scheme 4. A ring, Y, c, and Rare defined herein. cally in the presence of a trialkylamine. Such as triethylamine PG' and PG are amino protecting groups. Although the or diisopropylethylamine (DIEA). Suitable coupling reagents 30 schemes herein show R' and R as being hydrogen and a for use include, by way of example, carbodiimides, such as being one, the entire scope of R', R, and a as applicable to ethyl-3-(3-dimethylamino)propylcarbodiimide (EDC), dicy Formula I-XV is contemplated. clohexylcarbodiimide (DCC), diisopropylcarbodiimide (DIC) and the like, and other well-known coupling reagents, such as N,N'-carbonyldiimidazole, 2-ethoxy-1-ethoxycarbo 35 Scheme 4 nyl-1,2-dihydroquinoline (EEDQ), benzotriazol-1-yloxy-tris (dimethylamino)phosphonium hexafluorophosphate (BOP), HN YH-PG O-(7-azabenzotriazol-1-yl)-N.N.N.N',N'-tetramethyluro A -e- nium hexafluorophosphate (HATU) and the like. Optionally, well-known coupling promoters, such as N-hydroxySuccin 40 (Y), imide, 1-hydroxybenzotriazole (HOBT), 1-hydroxy-7-aza S-401 benzotriazole (HOAT), N,N-dimethylaminopyridine O (DMAP) and the like, can be employed in this reaction. Typi H cally, this coupling reaction is conducted at a temperature N-PG2 ranging from about 0°C. to about 60°C. for about 1 to about 45 HO 72 hours in an inert diluent, such as THF or DMF. In certain R5 instances, Compound S-301 reacts with Compound S-302 to N form Compound S-303 in the presence of HATU. HN NH2 S-403 In certain instances in Scheme 3, Compound S-301 is A -- reacted with Compound S-302 with R" as a protecting group 50 (Y), for an amino group. In these instances, the protecting group can be removed and the R group as an N-derivative group can S-402 be attached. Conditions for removal of other protecting O groups depend on the identity of the protecting group and are HN-PG? known to those skilled in the art. The conditions can also be 55 N NN found in Greene and Wuts. For example, a malonyl group can A H be attached via a reaction with mono-tert-butyl malonate. R5 Reaction using mono-tert-butyl malonate can be aided with (Y), use of activation reagents, such as symmetric anhydrides, O S-404 -(benzotriazol-1-yl)-N.N.N',N'-tetramethyluronium 60 hexafluorophosphate (HBTU), dicyclohexylcarbodiimide (DCC) diisopropylcarbodiimide (DIC)/1-hydroxybenzotria In Scheme 4, Compound S-401 is a commercially available Zole (HOBt), and benzotriazole-1-yl-oxytris(dimethy starting material. Alternatively, Compound S-401 can be lamino)phosphonium hexafluorophosphate (BOP). In semi-synthetically derived from natural materials or synthe another example, an alkanoyl group. Such as an acetyl group, 65 sized via a variety of different synthetic routes using com can be attached via a reaction with alkanoyl anhydride or mercially available starting materials and/or starting materi alkanoyl halide. als prepared by conventional synthetic methods. US 8,685,916 B2 73 74 With continued reference to Scheme 4, the protecting formates. In Scheme 5, the activation agent Compound S-502 group PG is removed from Compound S-401 to form Com is 4-nitrophenyl chloroformate. Other suitable activation pound S-402. Conditions to remove amino groups can be agents can be used prior to reaction with Compound S-404. found in Greene and Wuts. When PG' is a Boc group, the With continued reference to Scheme 5, activated Com protecting group can be removed with acidic conditions. Such pound S-501 reacts with Compound S-404 to form Com as treatment with hydrochloric acid or trifluoroacetic acid. pound S-503. In Scheme 5, Compound S-501 is a commer With continued reference to Scheme 4, Compound S-402 cially available starting material. Alternatively, Compound reacts with Compound S-403 to form Compound S-404 in a S-501 can be synthesized via a variety of different synthetic peptide coupling reaction. In certain embodiments, R is a routes using commercially available starting materials and/or side chain of an amino acid and is optionally protected. Pro 10 starting materials prepared by conventional synthetic meth tecting groups for the side chain of amino acids are known to ods. those skilled in art and can be found in Greene and Wuts. In certain instances, the protecting group for the side chain of arginine is a Sulfonyl-type protecting group. Such as 2.2.4.6. Scheme 6 7-pentamethyldihydrobenzofurane (Pbf). Other protecting 15 groups include 2.2.5.7.8-pentamethylchroman (Pmc) and O O 1,2-dimethylindole-3-sulfonyl (MIS). l N N-PG A peptide coupling reaction typically employs a conven A tional peptide coupling reagent and is conducted under con GoD-O RS N R5 -- ventional coupling reaction conditions, typically in the pres ence of a trialkylamine. Such as triethylamine or (Y), diisopropylethylamine (DIEA). Suitable coupling reagents for use include, by way of example, carbodiimides, such as S-503 ethyl-3-(3-dimethylamino)propylcarbodiimide (EDC), dicy O O clohexylcarbodiimide (DCC), diisopropylcarbodiimide 25 l N-R (DIC) and the like, and other well-known coupling reagents, such as N,N'-carbonyldiimidazole, 2-ethoxy-1-ethoxycarbo God-o15-yA H nyl-1,2-dihydroquinoline (EEDQ), benzotriazol-1-yloxy-tris R5 (dimethylamino)phosphonium hexafluorophosphate (BOP), (Y), O-(7-azabenzotriazol-1-yl)-N.N.N.N',N'-tetramethyluro 30 nium hexafluorophosphate (HATU) and the like. Optionally, S-601 well-known coupling promoters, such as N-hydroxySuccin imide, 1-hydroxybenzotriazole (HOBT), 1-hydroxy-7-aza benzotriazole (HOAT), N,N-dimethylaminopyridine Additional amino acids can be added to the compound (DMAP) and the like, can be employed in this reaction. Typi 35 through standard peptide coupling reactions as discussed cally, this coupling reaction is conducted at a temperature herein. For example, additional amino acids can be added to ranging from about 0°C. to about 60°C. for about 1 to about Compound S-503 with removal of protecting group PG and 72 hours in an inert diluent, such as THF or DMF. In certain addition of amino acids through standard peptide coupling instances, Compound S-402 reacts with Compound S-403 to reactions. Additional amino acids can be also added to Com 40 pound S-404 before reaction with Compound S-501 with form Compound S-404 in the presence of HATU. removal of protecting group PG and addition of amino acids A representative synthesis for Compound S-503 is shown through standard peptide coupling reactions. in Scheme 5. In Scheme 5. A ring, Y, c, and R are defined In Scheme 6, Compound S-503 is converted to Compound herein. PG is an amino protecting group. S-601 with R" as an N-derivative group. Conditions for 45 removal of protecting groups depend on the identity of the protecting group and are known to those skilled in the art. The Scheme 5 conditions can also be found in Greene and Wuts. In certain instances, for example, a malonyl group can be attached via a O Gpioid-OH reaction with mono-tert-butyl malonate. Reaction using N N-PG2 S-501 50 mono-tert-butyl malonate can be aided with use of activation N N O C reagents, such as symmetric anhydrides, O-(benzotriazol-1- A H yl)-N.N.N',N'-tetramethyluronium hexafluorophosphate R5 O (HBTU), dicyclohexylcarbodiimide (DCC) diisopropylcar (Y), ON bodiimide (DIC)/1-hydroxybenzotriazole (HOBt), and ben S-404 S-502 55 Zotriazole-1-yl-oxytris(dimethylamino)phosphonium O O hexafluorophosphate (BOP). In another example, an alkanoyl group, such as an acetyl group, can be attached via a reaction ul N-PG2 with alkanoyl anhydride or alkanoyl halide. Gioid-01 y-ryA H Removal of other protecting groups can be performed if R5 60 other protecting groups were used. Such as protecting groups (Y), present on the R or R moiety. Conditions for removal of other protecting groups depend on the identity of the protect S-503 ing group and are known to those skilled in the art. The conditions can also be found in Greene and Wuts. In Scheme 5, Compound S-501 is reacted with an activa 65 Enzyme Inhibitors tion agent, such as Compound S-502. Suitable activation The enzyme capable of cleaving the enzyme-cleavable agents include carbonate-forming reagents, such as chloro moiety of an opioid prodrug can be a peptidase, also called a US 8,685,916 B2 75 76 protease. In certain embodiments, the enzyme is an enzyme (phenylmethyl sulfonyl fluoride). Examples of small mol located in the gastrointestinal(GI) tract, i.e., a gastrointestinal ecule irreversible inhibitors that are protease inhibitors enzyme, or a GI enzyme. The enzyme can be a digestive include, but are not limited to benzamidine, apixaban, camo enzyme such as a gastric, intestinal, pancreatic or brush bor stat, 3,4-dichloroisocoumarin, e-aminocaprionic acid, der enzyme or enzyme of GI microbial flora, such as those amastatin, lysianadioic acid, 1,10-phenanthroline, cysteam involved in peptide hydrolysis. Examples include a pepsin, ine, and bestatin. Other examples of small molecule inhibi Such as pepsin A or pepsin B; a trypsin; a chymotrypsin; an tors are Compound 101, Compound 102, Compound 103. elastase; a carboxypeptidase, Such as carboxypeptidase A or Compound 104, Compound 105, Compound 106, Compound carboxypeptidase B; an aminopeptidase (such as aminopep 107, Compound 108, Compound 109 and Compound 110. tidase N or aminopeptidase A.; an endopeptidase; an exopep 10 The following table shows examples of gastrointestinal tidase; a dipeptidylaminopeptidase such as dipeptidylami nopeptidase IV: a dipeptidase; a tripeptidase; or an (GI) proteases, examples of their corresponding Substrates, enteropeptidase. In certain embodiments, the enzyme is a and examples of corresponding inhibitors. cytoplasmic protease located on or in the GI brush border. In certain embodiments, the enzyme is trypsin. Accordingly, in 15 certain embodiments, the corresponding composition is Table of Examples of GI Proteases and Corresponding Substrates and administered orally to the patient. Inhibitors The disclosure provides for a composition comprising a GI enzyme inhibitor. Such an inhibitor can inhibit at least one of GI Protease Substrates Inhibitors any of the GI enzymes disclosed herein. An example of a GI Trypsin Arg, LyS, TLCK, Benzamidine, enzyme inhibitor is a protease inhibitor, Such as a trypsin positively Apixaban, Bowman Birk inhibitor. charged residues Chymotrypsin Phe, Tyr, Trp, e-Aminocaprionic As used herein, the term “GI enzyme inhibitor” refers to bulky TPCK any agent capable of inhibiting the action of a GI enzyme on hydrophobic Bowman-Birk a substrate. The ability of an agent to inhibit a GI enzyme can 25 residues Pepsin Leu, Phe, Trp, Pepstatin, PMSF be measured using assays well known in the art. Tyr In certain embodiments, the GI enzyme capable of cleav Carboxypeptidase B Arg, Lys Potato-derived inhibitor, ing the enzyme-cleavable moiety may be a protease. The Lysianadioic acid disclosure provides for inhibitors of proteases. Carboxypeptidase A not Arg, Lys Potato-derived inhibitor, Proteases can be classified as exopeptidases or endopepti 30 1,10-phenanthroline Elastase Ala, Gly, Ser, C1-antitrypsin, 3,4- dases. Examples of exopeptidases include aminopeptidase Small neutral dichlorocoumarin and carboxypeptidase (A, B, or Y). Examples of endopepti residues dases include trypsin, chymotrypsin, elastase, pepsin, and Aminopeptidase All free N- Bestatin, Amastatin papain. The disclosure provides for inhibitors of exopepti terminal AA dase and endopeptidase. 35 In some embodiments, the enzyme is a digestive enzyme of Trypsin Inhibitors a protein. The disclosure provides for inhibitors of digestive As used herein, the term “trypsin inhibitor” refers to any enzymes. Agastric phase involves stomach enzymes, such as agent capable of inhibiting the action of trypsinona Substrate. pepsin. An intestinal phase involves enzymes in the Small The term “trypsin inhibitor also encompasses salts of trypsin intestine duodenum, Such as trypsin, chymotrypsin, elastase, 40 carboxypeptidase A, and carboxypeptidase B. An intestinal inhibitors. The ability of an agent to inhibit trypsin can be brush border phase involves enzymes in the small intestinal measured using assays well known in the art. For example, in brush border, Such as aminopeptidase N. aminopeptidase A, a typical assay, one unit corresponds to the amount of inhibi endopeptidases, dipeptidases, dipeptidylaminopeptidase, torthat reduces the trypsinactivity by one benzoyl-L-arginine and dipeptidylaminopeptidase IV. An intestinal intracellular 45 ethyl ester unit (BAEE-U). One BAEE-U is the amount of phase involves intracellular peptidases, such as dipeptidases enzyme that increases the absorbance at 253 nm by 0.001 per (i.e. iminopeptidase) and aminopeptidase. minute at pH 7.6 and 25°C. See, for example, K. Ozawa, M. In certain embodiments, the enzyme inhibitor in the dis Laskowski, 1966, J. Biol. Chem. 241,3955 andY. Birk, 1976, closed compositions is a peptidase inhibitor or protease Meth. Enzymol. 45,700. In certain instances, a trypsin inhibi inhibitor. In certain embodiments, the enzyme is a digestive 50 enzyme Such as a gastric, pancreatic or brush border enzyme, tor can interact with an active site of trypsin, Such as the S1 Such as those involved in peptide hydrolysis. Examples pocket and the S3/4 pocket. The S1 pocket has an aspartate include pepsin, trypsin, chymotrypsin, colipase, elastase, residue which has affinity for positively charged moiety. The aminopeptidase N. aminopeptidase A, dipeptidylaminopep S3/4 pocket is a hydrophobic pocket. The disclosure provides tidase IV, tripeptidase or enteropeptidase. 55 for specific trypsin inhibitors and non-specific serine protease Proteases can be inhibited by naturally occurring peptide inhibitors. or protein inhibitors, or by Small molecule naturally occurring There are many trypsin inhibitors known in the art, both or synthetic inhibitors. Examples of protein or peptide inhibi those specific to trypsin and those that inhibit trypsin and tors that are protease inhibitors include, but are not limited to, other proteases such as chymotrypsin. The disclosure pro al-antitrypsin from human plasma, aprotinin, trypsin inhibi 60 vides for trypsin inhibitors that are proteins, peptides, and tor from soybean (SBTI), Bowman-Birk Inhibitor from soy bean (BBSI), trypsin inhibitor from egg white (ovomucoid), small molecules. The disclosure provides for trypsin inhibi chromostatin, and potato-derived carboxypeptidase inhibitor. tors that are irreversible inhibitors or reversible inhibitors. Examples of small molecule irreversible inhibitors that are The disclosure provides for trypsin inhibitors that are com protease inhibitors include, but are not limited to, TPCK 65 petitive inhibitors, non-competitive inhibitors, or uncompeti (1-chloro-3-tosylamido-4-phenyl-2-butanone), TLCK tive inhibitors. The disclosure provides for natural, synthetic (1-chloro-3-tosylamido-7-amino-2-heptone), and PMSF or semi-synthetic trypsin inhibitors. US 8,685,916 B2 77 78 Trypsin inhibitors can be derived from a variety of animal wherein: or vegetable sources: for example, soybean, corn, lima and Q is selected from O-Q or -Q-COOH, where Q is other beans, squash, Sunflower, bovine and other animal pan C-C alkyl; creas and lung, chicken and turkey egg white, soy-based infantformula, and mammalian blood. Trypsin inhibitors can 5 Q is N or CH; and also be of microbial origin: for example, antipain; see, for Q is aryl or substituted aryl. example, H. Umezawa, 1976, Meth. Enzymol. 45,678. Certain trypsin inhibitors include compounds of formula: In one embodiment, the trypsin inhibitor is derived from soybean. Trypsin inhibitors derived from soybean (Glycine 10 max) are readily available and are considered to be safe for (Q), NH2, human consumption. They include, but are not limited to, r SBTI, which inhibits trypsin, and Bowman-Birk inhibitor, Q3 NH which inhibits trypsin and chymotrypsin. Such trypsin inhibi- pi tors are available, for example from Sigma-Aldrich, St. 15 Louis, Mo., USA. A trypsin inhibitor can be an arginine mimic or lysine wherein: mimic, either natural or synthetic compound. In certain Q is –C(O)-COOH or - NH-Q-Q-SO CHs. embodiments, the trypsin inhibitor is an arginine mimic or a where lysine mimic, wherein the arginine mimic O lysine mimic is 20 Q is -(CH2). COOH: a synthetic compound. As used herein, an arginine mimic or 7 lysine mimic can include a compound capable of binding to Q is —(CH2), CHs; the P' pocket of trypsin and/or interfering with trypsin active Q is NH; site function. The arginine or lysine mimic can be a cleavable as n is a number from Zero to two: or non-cleavable moiety. Examples of trypsin inhibitors,- - - - which are arginine mimics o 1S Zero or one; and/or lysine mimics, include, but not limited to, arylguani- p is an integer from one to three; and dine, benzamidine, 3,4-dichloroisocoumarin, diisopropy- r is an integer from one to three. ifluorophosphate, gabexate mesylate, and phenylmethane- 30 Other examples of trypsin inhibitors include compounds of sulfonyl fluoride, or substituted versions or analogs thereof. formula: In certain embodiments, trypsin inhibitors comprise a covalently modifiable group. Such as a chloroketone moiety, an aldehyde moiety, or an epoxide moiety. Other examples of NH trypsin inhibitors are aprotinin, camo.stat and pentamidine. 35 Other examples of trypsin inhibitors include compounds of NH2, formula: Q3

Q1 O 40 4. Y. -Q3 wherein: O 1. HN \, Q is –C(O)-COOH or - NH-Q-Q7-SO, CH, N- where H 45 Q is —(CH.). COOH: O N NH2, Q is —(CH), C.Hs; and p is an integer from one to three; and NH r is an integer from one to three. Certain trypsin inhibitors include the following:

Compound (S)-ethyl 4-(5-guanidino 101 2-(naphthalene-2- Sulfonamido)pentanoyl) O O piperazine-1-carboxylate O1. r HN1W\, N O

US 8,685,916 B2 83 84 A description of methods to prepare Compound 101, Com represents a 4-7 membered monocyclic hetero-ring contain pound 102, Compound 103, Compound 104, Compound 105, ing 1 to 2 nitrogen or oxygen atoms, Compound 107, and Compound 108 is provided in PCT Inter R'' represents a hydrogenatom, a C- alkyl group substi national Publication Number WO 2010/045599A1, pub tuted by a phenyl group or a group of formula: COOR'7. lished 22 Apr. 2010, which is hereby incorporated by refer wherein R' represents a hydrogen atom, a C- alkyl group ence in its entirety. Compound 106, Compound 109, and or a C alkyl group Substituted by a phenyl group; Compound 110 can be obtained commercially (Sigma-Ald rich, St. Louis, Mo., USA.). provided that R', R'' and R' do not represent simulta In certain embodiments, the trypsin inhibitor is SBTI, neously hydrogen atoms; BBSI, Compound 101, Compound 106, Compound 108, 10 or nontoxic salts, acid addition salts or hydrates thereof. Compound 109, or Compound 110. In certain embodiments, In certain embodiments, the trypsin inhibitor is a com the trypsin inhibitor is camoStat. pound selected from the following: In certain embodiments, the trypsin inhibitor is a com pound of formula T-I: 15

(T-I) O o

HN A. HN COOCHs HC O HN O o wherein 25 A represents a group of the following formula: HNX--()-d KCOOCH5 HN O M s COOCHs, or -CEC A R ig Riv,i0 30 cy-O RS R’ and R' each represents independently a hydrogen atom or a C alkyl group, R represents a group selected from the following formu 35 HN ) () , lae: )--()–: -Uy

Rt 11 Rt 11 In certain embodiments, the trypsin inhibitor is a com CON CON 40 pound of formula T-II: Rt 12 Rt-( 13 Rt 12 O cC (T-II) wherein R', R'' and R' each represents independently HN (1) a hydrogen atom, 45 (2) a phenyl group, HNX- X -()– Ril (3) a C alkyl group Substituted by a phenyl group, (4) a Co alkyl group, (5) a Coalkoxyl group, wherein (6) a Coalkenyl group having 1 to 3 double bonds, 50 X is NH; (7) a Co alkynyl group having 1 to 2 triple bonds, n is Zero or one; and (8) a group of formula: R' C(O)xR', R' is selected from hydrogen, halogen, nitro, alkyl, Sub wherein R' represents a single bond or a Cls alkylene stituted alkyl, alkoxy, carboxyl, alkoxycarbonyl, acyl, ami grOup, 55 noacyl, guanidine, amidino, carbamide, amino, Substituted X represents an oxygen atom or an NH-group, and amino, hydroxyl, cyano and —(CH), C(O)—O— R' represents a hydrogen atom, a C- alkyl group, a (CH), C(O) N. R'R'', wherein each m is indepen phenyl group or a C alkyl group Substituted by a dently zero to 2; and R" and R' are independently selected phenyl group, or from hydrogen and C alkyl. (9) a C-7 cycloalkyl group: 60 the structure In certain embodiments, informula T-II, R' is guanidino or amidino. In certain embodiments, in formula T-II, R' is (CH), C(O)-O-(CH), C(O)-N-R'R'', S) 65 wherein m is one and R'' and R' are methyl. In certain embodiments, the trypsin inhibitor is a com pound of formula T-III: US 8,685,916 B2 85 86 In certain embodiments, the trypsin inhibitor is (T-III) HN O 5 HNX-x-( )--- O ouls N1 O NH O wherein

X is NH; 10 H2 -- H n is Zero or one; L' is selected from C(O)—O : —O C(O)— —O In certain embodiments, the trypsin inhibitor is Compound (CH), O—: OCH Art CHO : C(O) 110 or a bis-arylamidine variant thereof; see, for example, J. NR' ; and NR. C(O) ; 15 D. Geratz, M. C.-F. Cheng and R. R. Tidwell (1976) J. Med. R’ is selected from hydrogen, C alkyl, and substituted Chem. 19, 634–639. C. alkyl, It will be appreciated that the pharmaceutical composition Ar' and Arfare independently a substituted or unsubsti according to the embodiments may further comprise one or tuted aryl group; more additional trypsin inhibitors. It is to be appreciated that the invention also includes m is a number from 1 to 3; and inhibitors of other enzymes involved in protein assimilation R is selected from hydrogen, halogen, nitro, alkyl, Sub that can be used in combination with a prodrug disclosed stituted alkyl, alkoxy, carboxyl, alkoxycarbonyl, acyl, ami herein comprising an amino acid of alanine, arginine, aspar noacyl, guanidine, amidino, carbamide, amino, Substituted agine, aspartic acid, cysteine, glutamic acid, glutamine, gly amino, hydroxyl, cyano and —(CH), C(O)—O— 25 cine, histidine, isoleucine, leucine, lysine, methionine, phe (CH), C(O) N. R'R'', wherein each m is indepen nylalanine, proline, serine, threonine, tryptophan, tyrosine, or dently zero to 2; and R'' and R' are independently selected Valine or amino acid variants thereof. from hydrogen and C alkyl. Combinations of Prodrug and Trypsin Inhibitor As disclosed above, the present disclosure also provides In certain embodiments, in formula T-III, R is guanidino 30 pharmaceutical compositions, and their methods of use, or amidino. where the pharmaceutical compositions comprise an opioid In certain embodiments, in formula T-III, R' is prodrug, that provides controlled release of an opioid, and a (CH), C(O)-O-(CH), C(O)-N-R'R'', trypsin inhibitor that interacts with the trypsin that mediates wherein m is one and R'' and R" are methyl. the-controlled release of the opioid from the prodrug so as to 35 attenuate enzymatic cleavage of the prodrug. In certain embodiments, the trypsin inhibitor is a com The embodiments provide a pharmaceutical composition, pound of formula T-IV: which comprises a trypsin inhibitor and a compound of gen eral Formulae I-XII, or a pharmaceutically acceptable salt thereof. The embodiments provide a pharmaceutical compo (T-IV) 40 sition, which comprises a trypsin inhibitor and a compound of NH general Formulae XIII-XV, or a pharmaceutically acceptable HN salt thereof. The embodiments provide a pharmaceutical composition, which comprises a compound of Formulae T-I to T-IV and a HN}-x-( )--v-x. NH2 45 compound of general Formulae I-XII, or a pharmaceutically acceptable salt thereof. The embodiments provide a pharma ceutical composition, which comprises a compound of For wherein mulae T-I to T-IV and a compound of general Formulae each X is NH; XIII-XV, or a pharmaceutically acceptable salt thereof. The each n is independently Zero or one; 50 embodiments provide a pharmaceutical composition, which comprises Compound 109 and a compound of general For L' is selected from —C(O)—O : —O C(O)— —O mulae I-XII, or a pharmaceutically acceptable salt thereof. (CH), O—; OCH Ar’ CHO : C(O) The embodiments provide a pharmaceutical composition, NR' ; and NR. C(O) ; which comprises Compound 109 and a compound of general R is selected from hydrogen, C. alkyl, and substituted 55 Formulae XIII-XV, or a pharmaceutically acceptable salt C. alkyl, thereof. Ar' and Arfare independently a substituted or unsubsti Certain embodiments provide for a combination of a com tuted aryl group; and pound of Formula I and a trypsin inhibitor, shown in the table below. Certain embodiments provide for a combination of a m is a number from 1 to 3. 60 compound of Formulae II-V and a trypsin inhibitor, shown in In certain embodiments, in formula T-IV. Ar' or Ar' is the table below. Certain embodiments provide for a combi phenyl. nation of a compound of Formulae VI-IX and a trypsin inhibi In certain embodiments, in formula T-IV. Ar' or Ar' is tor, shown in the table below. Certain embodiments provide naphthyl. for a combination of a compound of Formulae X-XII and a 65 trypsin inhibitor, shown in the table below. Certain embodi In certain embodiments, the trypsin inhibitor is Compound ments provide for a combination of a compound of Formulae 109. XIII-XV and a Trypsin inhibitor, shown in the table below. US 8,685,916 B2 87 88

Prodrug of Formula Prodrug of Formula Prodrug of Formula Prodrug of Formula Prodrug of Formula I II-V and Trypsin VI-IX and Trypsin X-XII and Trypsin XIII-XV and Trypsin and Trypsin Inhibitor Inhibitor Inhibitor Inhibitor Inhibitor SBTI SBTI SBTI SBTI SBTI BBSI BBSI BBSI BBSI BBSI Compound 101 Compound 101 Compound 101 Compound 101 Compound 101 Compound 106 Compound 106 Compound 106 Compound 106 Compound 106 Compound 108 Compound 108 Compound 108 Compound 108 Compound 108 Compound 109 Compound 109 Compound 109 Compound 109 Compound 109 Compound 110 Compound 110 Compound 110 Compound 110 Compound 110

Combinations of Opioid Prodrugs and Other Drugs use in therapy or prevention or as a medicament. The present The disclosure provides for an opioid prodrug of the 15 disclosure also provides the use of a disclosed composition embodiments and a further prodrug or drug included in a for the manufacture of a medicament, especially for the pharmaceutical composition. Such a prodrug or drug would manufacture of a medicament for the treatment or prevention provide additional analgesia, e.g., a synergistic effect, or of pain. other benefits. Examples include opioids, opioid prodrugs, The compositions of the present disclosure can be used in acetaminophen, non-steroidal anti-inflammatory drugs the treatment or prevention of pain including, but not limited (NSAIDs) and other analgesics. In one embodiment, two or to include, acute pain, chronic pain, neuropathic pain, acute more opioid agonist prodrugs and/or drugs (e.g., a morphine traumatic pain, arthritic pain, osteoarthritic pain, rheumatoid prodrug or drug and an oxycodone prodrug of the embodi arthritic pain, muscular skeletal pain, post-dental Surgical ments), each at a Sub-analgesic dose, would be combined to pain, dental pain, myofascial pain, cancer pain, visceral pain, provide a synergistic response leading to effective analgesia 25 diabetic pain, muscular pain, post-herpetic neuralgic pain, with reduced side effects. In one embodiment, an opioid chronic pelvic pain, endometriosis pain, pelvic inflammatory agonist prodrug or drug would be combined with an opioid pain and childbirth related pain. Acute pain includes, but is antagonist prodrug or drug. Other examples include drugs or not limited to, acute traumatic pain or post-Surgical pain. prodrugs that have benefits other than, or in addition to, Chronic pain includes, but is not limited to, neuropathic pain, analgesia. The embodiments provide a pharmaceutical com 30 arthritic pain, osteoarthritic pain, rheumatoid arthritic pain, position, which comprises an opioid prodrug and acetami muscular skeletal pain, dental pain, myofascial pain, cancer nophen and optionally comprises an enzyme inhibitor. Also pain, diabetic pain, visceral pain, muscular pain, post-her included are pharmaceutically acceptable salts thereof. petic neuralgic pain, chronic pelvic pain, endometriosis pain, In certain embodiments, the enzyme inhibitor is selected pelvic inflammatory pain and back pain. from SBTI, BBSI, Compound 101, Compound 106, Com 35 The present disclosure provides use of a compound of pound 108, Compound 109, and Compound 110. In certain Formulae I-XV in the treatment of pain. The present disclo embodiments, the enzyme inhibitor is camoStat. sure provides use of a compound of Formulae I-XV in the In certain embodiments, a pharmaceutical composition can prevention of pain. comprise an opioid prodrug, a non-opioid drug and at least The present disclosure provides use of a compound of one opioid or opioid prodrug. 40 Formulae I-XV in the manufacture of a medicament for treat Pharmaceutical Compositions and Methods of Use ment of pain. The present disclosure provides use of a com The present disclosure provides a composition, such as a pound of Formulae I-XV in the manufacture of a medicament pharmaceutical composition, which comprises a compound for prevention of pain. of Formulae I-XV. Such a pharmaceutical composition In another aspect, the embodiments provide a method of according to the embodiments can further comprise a phar 45 treating pain in a patient in need thereof, which comprises maceutically acceptable carrier. The composition is conve administering to Such a patient an effective amount of a phar niently formulated in a form suitable for oral (including buc maceutical composition as described herein. In another cal and Sublingual) administration, for example as a tablet, aspect, the embodiments provide a method of preventing pain capsule, thin film, powder, Suspension, Solution, syrup, dis in a patient in need thereof, which comprises administering to persion or emulsion. The composition can contain compo 50 Such a patient an effective amount of a pharmaceutical com nents conventional in pharmaceutical preparations, e.g. one position as described herein. or more carriers, binders, lubricants, excipients (e.g., to The amount of composition disclosed herein to be admin impart controlled release characteristics), pH modifiers, istered to a patient to be effective (i.e. to provide blood levels Sweeteners, bulking agents, coloring agents or further active of an opioid sufficient to be effective in the treatment or agents. 55 prophylaxis of pain) will depend upon the bioavailability of Patients can be humans, and also other mammals, such as the particular composition, the Susceptibility of the particular livestock, Zoo animals and companion animals. Such as a cat, composition to enzyme activation in the gut, as well as other dog or horse. factors, such as the species, age, weight, sex, and condition of In another aspect, the embodiments provide a pharmaceu the patient, manner of administration and judgment of the tical composition as described herein for use in the treatment 60 prescribing physician. If the composition also comprises a of pain. The pharmaceutical composition according to the trypsin inhibitor, the amount of composition disclosed herein embodiments is useful, for example, in the treatment of a to be administered to a patient would also depend on the patient Suffering from, or at risk of Suffering from pain. amount and potency of trypsin inhibitor present in the com Accordingly, the present disclosure provides methods of position. In general, the dose can be such that the opioid treating or preventing pain in a Subject, the methods involving 65 prodrug is in the range of from 0.01 milligrams per kilogram administering to the Subject a disclosed composition. The to 20 milligrams prodrug per kilogram (mg/kg) body weight. present disclosure provides for a disclosed composition for For example, an opioid prodrug can be administered at a dose US 8,685,916 B2 89 90 equivalent to administering free opioid in the range of from The embodiments include a dose unit comprising a com 0.02-mg/kg to 0.5-mg/kg body weight or 0.01-mg/kg to position, such as a pharmaceutical composition, comprising 10-mg/kg body weight or 0.01-mg/kg to 2-mg/kg body an opioid prodrug of Formulae I-XV and a GI enzyme inhibi weight. In one embodiment, the composition can be admin tor, where the opioid prodrug of Formulae I-XV and GI istered at a dose such that the level of the opioid achieved in enzyme inhibitor are present in the dose unit in an amount the blood is in the range of from 0.5 ng/ml to 200 ng/ml. In one effective to provide for a pre-selected pharmacokinetic (PK) embodiment, the composition can be administered at a dose profile following ingestion. In further embodiments, the pre such that the level of the opioid achieved in the blood is in the selected PK profile comprises at least one PK parameter value range of from 0.5 ng/ml to 20 ng/ml. In one embodiment, the that is less than the PK parameter value of opioid released composition can be administered at a dose such that the level 10 following ingestion of an equivalent dosage of an opioid of the opioid achieved in the blood is in the range of from 0.5 ng/ml to 10 ng/ml. prodrug of Formulae I-XV in the absence of inhibitor. In As disclosed above, the present disclosure also provides a further embodiments, the PK parameter value is selected pharmaceutical composition that comprises an opioid pro from an opioid Cmax value, an opioid exposure value, and a drug of Formulae I-XV and a trypsin inhibitor. Such an opioid 15 (1/opioid Tmax) value. prodrug comprises a promoiety comprising a trypsin-cleav In certain embodiments, the dose unit provides for a pre able moiety that, when cleaved, facilitates release of opioid. selected PK profile following ingestion of at least two dose The present disclosure provides use of a compound of units. In related embodiments, the pre-selected PK profile of Formulae I-XV and a trypsin inhibitor in the treatment of such dose units is modified relative to the PK profile follow pain. The present disclosure provides use of a compound of ing ingestion of an equivalent dosage of an opioid prodrug of Formulae I-XV and a trypsin inhibitor in the prevention of Formulae I-XV without inhibitor. In related embodiments, pa1n. Such a dose unit provides that ingestion of an increasing The present disclosure provides use of a compound of number of the dose units provides for a linear PK profile. In Formulae I-XV and a trypsin inhibitor in the manufacture of related embodiments. Such a dose unit provides that ingestion a medicament for treatment of pain. The present disclosure 25 of an increasing number of the dose units provides for a provides use of a compound of Formulae I-XV and a trypsin nonlinear PK profile. In related embodiments, the PK param inhibitor in the manufacture of a medicament for prevention eter value of the PK profile of such a dose unit is selected from of pain. an opioid Cmax value, a (1/opioid Tmax) value, and an opioid In another aspect, the embodiments provide a method of exposure value. treating pain in a patient in need thereof, which comprises 30 The embodiments include methods for treating a patient administering to Such a patient an effective amount of a phar comprising administering any of the compositions, such as maceutical composition comprising a compound of Formulae pharmaceutical compositions, comprising an opioid prodrug I-XV and a trypsin inhibitor. In another aspect, the embodi of Formulae I-XV and a GI enzyme inhibitor or dose units ments provide a method of preventing pain inapatient in need described herein to a patient in need thereof. The embodi thereof, which comprises administering to such a patient an 35 ments include methods to reduce side effects of a therapy effective amount of a pharmaceutical composition compris comprising administering any of Such compositions, e.g., ing a compound of Formulae I-XV and a trypsin inhibitor. pharmaceutical compositions, or dose units described herein, In Such pharmaceutical compositions, the amount of a to a patient in need thereof. The embodiments include meth trypsin inhibitor to be administered to the patient to be effec ods of improving patient compliance with a therapy pre tive (i.e. to attenuate release of an opioid when administration 40 scribed by a clinician comprising directing administration of of a compound disclosed herein alone would lead to overex any of such compositions, e.g., pharmaceutical compositions, posure of the opioid) will depend upon the effective dose of or dose units described herein, to a patient in need thereof. the particular prodrug and the potency of the particular inhibi Such embodiments can provide for improved patient compli tor, as well as other factors, such as the species, age, weight, ance with a prescribed therapy as compared to patient com sex and condition of the patient, manner of administration and 45 pliance with a prescribed therapy using drug and/or using judgment of the prescribing physician. In general, the dose of prodrug without inhibitor as compared to prodrug with inhibitor can be in the range of from 0.05 mg to 50 mg per mg inhibitor. of prodrug disclosed herein. In a certain embodiment, the The embodiments include methods of reducing risk of dose of inhibitor can be in the range of from 0.001 mg to 50 unintended overdose of an opioid comprising directing mg per mg of prodrug disclosed herein. In one embodiment, 50 administration of any of such compositions, e.g., pharmaceu the dose of inhibitor can be in the range of from 0.01 nano tical compositions, or dose units described herein, to a patient moles to 100 micromoles per micromole of prodrug disclosed in need of treatment. herein. The embodiments include methods of making a dose unit Representative Embodiments of Dose Units of Prodrug and comprising combining an opioid prodrug of Formulae I-XV GI Enzyme Inhibitor Having a Desired Pharmacokinetic Pro 55 and a GI enzyme inhibitor in a dose unit, wherein the opioid file prodrug of Formulae I-XV and GI enzyme inhibitor are The embodiments include a composition that comprises (a) present in the dose unit in an amount effective to attenuate an opioid prodrug of Formulae I-XV, which comprises an release of opioid from the opioid prodrug of Formulae I-XV. opioid covalently bound to a promoiety comprising a GI The embodiments include methods of deterring misuse or enzyme-cleavable moiety, wherein cleavage of the GI 60 abuse of multiple dose units of an opioid prodrug of Formulae enzyme-cleavable moiety by a GI enzyme mediates release of I-XV comprising combining an opioid prodrug of Formulae an opioid, and (b) a GI enzyme inhibitor that interacts with the I-XV and a GI enzyme inhibitor in a dose unit, wherein the GI enzyme that mediates enzymatically-controlled release of opioid prodrug of Formulae I-XV and GI enzyme inhibitor the opioid from the prodrug following ingestion of the com are present in the dose unit in an amount effective to attenuate position. In one embodiment, the GI enzyme is trypsin, the GI 65 release of an opioid from the opioid prodrug of Formulae enzyme-cleavable moiety is a trypsin-cleavable moiety, and I-XV such that ingestion of multiples of dose units by a the GI enzyme inhibitor is a trypsin inhibitor. patient does not provide a proportional release of the opioid. US 8,685,916 B2 91 92 In further embodiments, release of drug is decreased com therapeutic range). “Multiple dose units’ or “multiples of a pared to release of drug by an equivalent dosage of prodrug in dose unit' or a “multiple of a dose unit” refers to at least two the absence of inhibitor. single dose units. One embodiment is a method for identifying a GI enzyme As used herein, a “PK profile' refers to a profile of drug inhibitor and prodrug of Formulae I-XV suitable for formu concentration in blood or plasma. Such a profile can be a lation in a dose unit. Such a method can be conducted as, for relationship of drug concentration over time (i.e., a “concen example, an in vitro assay, an in vivo assay, or an ex vivo tration-time PK profile') or a relationship of drug concentra assay. In one embodiment, the GI enzyme inhibitor is a tion versus number of doses ingested (i.e., a “concentration trypsin inhibitor. dose PK profile'.) A PK profile is characterized by PK 10 parameters. The embodiments include methods for identifying a GI As used herein, a “PK parameter” refers to a measure of enzyme inhibitor and prodrug of Formulae I-XV suitable for drug concentration in blood or plasma, such as: 1) "drug formulation in a dose unit comprising combining a prodrug of Cmax', the maximum concentration of drug achieved in Formulae I-XV, a GI enzyme inhibitor, and a GI enzyme in a blood or plasma; 2) "drug Tmax', the time elapsed following reaction mixture, and detecting prodrug conversion, wherein 15 ingestion to achieve Cmax; and 3) "drug exposure', the total a decrease in prodrug conversion in the presence of the GI concentration of drug present in blood or plasma over a enzyme inhibitor as compared to prodrug conversion in the selected period of time, which can be measured using the area absence of the GI enzyme inhibitor indicates the GI enzyme under the curve (AUC) of a time course of drug release over inhibitor and prodrug of Formulae I-XV are suitable for for a selected period of time (t). Modification of one or more PK mulation in a dose unit. parameters provides for a modified PK profile. The embodiments include methods for identifying a GI For purposes of describing the features of dose units of the enzyme inhibitor and prodrug of Formulae I-XV suitable for present disclosure, “PK parameter values” that define a PK formulation in a dose unit comprising administering to an profile include drug Cmax (e.g., opioid Cmax), total drug animala GI enzyme inhibitor and prodrug of Formulae I-XV exposure (e.g., area under the curve) (e.g., opioid exposure) and detecting prodrug conversion, wherein a decrease in 25 and 1/(drug Tmax) (such that a decreased 1/Tmax is indica opioid conversion in the presence of the GI enzyme inhibitor tive of a delay in Tmax relative to a reference Tmax) (e.g., as compared to opioid conversion in the absence of the GI 1/opioid Tmax). Thus a decrease in a PK parameter value enzyme inhibitor indicates the GI enzyme inhibitor and pro relative to a reference PK parameter value can indicate, for drug of Formulae I-XV are suitable for formulation in a dose example, a decrease in drug Cmax, a decrease in drug expo unit. In certain embodiments, administering comprises 30 Sure, and/or a delayed Tmax. Dose units of the present disclosure can be adapted to administering to the animal increasing doses of inhibitor co provide for a modified PK profile, e.g., a PK profile that is dosed with a selected fixed dose of prodrug. Detecting pro different from that achieved from dosing a given dose of drug conversion can facilitate identification of a dose of prodrug in the absence of inhibitor (i.e., without inhibitor). inhibitor and a dose of prodrug that provides for a pre-se 35 For example, dose units can provide for at least one of lected pharmacokinetic (PK) profile. Such methods can be decreased drug Cmax, delayed drug Tmax and/or decreased conducted as, for example, an in vivo assay or an ex vivo drug exposure compared to ingestion of a dose of prodrug in assay. the same amount but in the absence of inhibitor. Such a The embodiments include methods for identifying a GI modification is due to the inclusion of an inhibitor in the dose enzyme inhibitor and prodrug of Formulae I-XV suitable for 40 unit. formulation in a dose unit comprising administering to an As used herein, “a pharmacodynamic (PD) profile' refers animal tissue a GI enzyme inhibitor and prodrug of Formulae to a profile of the efficacy of a drug in a patient (or Subject or I-XV and detecting prodrug conversion, wherein a decrease user), which is characterized by PD parameters. “PD param in prodrug conversion in the presence of the GI enzyme eters' include “drug Emax” (the maximum drug efficacy), inhibitor as compared to prodrug conversion in the absence of 45 “drug EC50' (the concentration of drug at 50% of the Emax), the GI enzyme inhibitor indicates the GI enzyme inhibitor and side effects. and prodrug of Formulae I-XV are suitable for formulation in FIG. 1 is a schematic illustrating an example of the effect of a dose unit. increasing inhibitor concentrations upon the PK parameter Dose Units of Prodrug and Inhibitor Having a Desired Phar drug Cmax for a fixed dose of prodrug. At low concentrations macokinetic Profile 50 of inhibitor, there may be no detectable effect on drug release, The present disclosure provides dose units of prodrug and as illustrated by the plateau portion of the plot of drug Cmax inhibitor that can provide for a desired pharmacokinetic (PK) (Y axis) versus inhibitor concentration (X axis). As inhibitor profile. Dose units can provide a modified PK profile com concentration increases, a concentration is reached at which pared to a reference PK profile as disclosed herein. It will be drug release from prodrug is attenuated, causing a decrease appreciated that a modified PK profile can provide for a 55 in, or suppression of drug Cmax. Thus, the effect of inhibitor modified pharmacodynamic (PD) profile. Ingestion of mul upon a prodrug PK parameter for a dose unit of the present tiples of such a dose unit can also provide a desired PK profile. disclosure can range from undetectable, to moderate, to com Unless specifically stated otherwise, “dose unit as used plete inhibition (i.e., no detectable drug release). herein refers to a combination of a GI enzyme-cleavable A dose unit can be adapted to provide for a desired PK prodrug (e.g., trypsin-cleavable prodrug) and a GI enzyme 60 profile (e.g., a concentration-time PK profile) following inhibitor (e.g., a trypsin inhibitor). A 'single dose unit' is a ingestion of a single dose. A dose unit can be adapted to single unit of a combination of a GI enzyme-cleavable pro provide for a desired PK profile (e.g., a concentration-dose drug (e.g., trypsin-cleavable prodrug) and a GI enzyme PK profile) followingingestion of multiple dose units (e.g., at inhibitor (e.g., trypsin inhibitor), where the single dose unit least 2, at least 3, at least 4 or more dose units). provide a therapeutically effective amount of drug (i.e., a 65 Dose Units Providing Modified PK Profiles Sufficient amount of drug to effect a therapeutic effect, e.g., a A combination of a prodrug and an inhibitor in a dose unit dose within the respective drugs therapeutic window, or can provide a desired (or “pre-selected') PK profile (e.g., a US 8,685,916 B2 93 94 concentration-time PK profile) following ingestion of a example, dose units with a decreased drug Cmax provide less single dose. The PK profile of such a dose unit can be char reward for ingestion than does a dose of the same amount of acterized by one or more of a pre-selected drug Cmax, a drug, and/or the same amount of prodrug without inhibitor. pre-selected drug Tmax or a pre-selected drug exposure. The Dose Units Providing Modified Pk Profiles Upon Ingestion PK profile of the dose unit can be modified compared to a PK of Multiple Dose Units profile achieved from the equivalent dosage of prodrug in the A dose unit of the present disclosure can be adapted to absence of inhibitor (i.e., a dose that is the same as the dose provide for a desired PK profile (e.g., a concentration-time unit except that it lacks inhibitor). PK profile or concentration-dose PK profile) following inges A modified PK profile can have a decreased PK parameter tion of multiples of a dose unit (e.g., at least 2, at least 3, at value relative to a reference PK parameter value (e.g., a PK 10 least 4, or more dose units). A concentration-dose PK profile parameter value of a PK profile following ingestion of a refers to the relationship between a selected PK parameter dosage of prodrug that is equivalent to a dose unit except and a number of single dose units ingested. Such a profile can without inhibitor). For example, a dose unit can provide for a be dose proportional, linear (a linear PK profile) or nonlinear decreased drug Cmax, decreased drug exposure, and/or (a nonlinear PK profile). A modified concentration-dose PK delayed drug Tmax. 15 profile can be provided by adjusting the relative amounts of FIG. 2 presents schematic graphs showing examples of prodrug and inhibitor contained in a single dose unit and/or by modified concentration-time PK profiles of a single dose unit. using a different prodrug and/or inhibitor. Panel A is a schematic of drug concentration in blood or FIG. 3 provides schematics of examples of concentration plasma (Y axis) following a period of time (X axis) after dose PK profiles (exemplified by drug Cmax,Y axis) that can ingestion of prodrug in the absence or presence of inhibitor. be provided by ingestion of multiples of a dose unit (X axis) The solid, top line in Panel A provides an example of drug of the present disclosure. Each profile can be compared to a concentration following ingestion of prodrug without inhibi concentration-dose PK profile provided by increasing doses tor. The dashed, lower line in Panel A represents drug con of drug alone, where the amount of drug in the blood or centration following ingestion of the same dose of prodrug plasma from one dose represents a therapeutically effective with inhibitor. Ingestion of inhibitor with prodrug provides 25 amount equivalent to the amount of drug released into the for a decreased drug Cmax relative to the drug Cmax that blood or plasma by one dose unit of the disclosure. Such a results from ingestion of the same amount of prodrug in the “drug alone. PK profile is typically dose proportional, having absence of inhibitor. Panel A also illustrates that the total drug a forty-five degree angle positive linear slope. It is also to be exposure following ingestion of prodrug with inhibitor is also appreciated that a concentration-dose PK profile resulting decreased relative to ingestion of the same amount of prodrug 30 fromingestion of multiples of a dose unit of the disclosure can without inhibitor. also be compared to other references, such as a concentration Panel B of FIG. 2 provides another example of a dose unit dose PK profile provided by ingestion of an increasing num having a modified concentration-time PK profile. As in Panel ber of doses of prodrug without inhibitor wherein the amount A, the Solid top line represents drug concentration over time of drug released into the blood or plasma by a single dose of in blood or plasma following ingestion of prodrug without 35 prodrug in the absence of inhibitor represents a therapeuti inhibitor, while the dashed lower line represents drug concen cally effective amount equivalent to the amount of drug tration following ingestion of the same amount of prodrug released into the blood or plasma by one dose unit of the with inhibitor. In this example, the dose unit provides a PK disclosure. profile having a decreased drug Cmax, decreased drug expo As illustrated by the relationship between prodrug and Sure, and a delayed drug Tmax (i.e., decreased (1/drug Tmax) 40 inhibitor concentration in FIG. 2, a dose unit can include relative to ingestion of the same dose of prodrug without inhibitor in an amount that does not detectably affect drug inhibitor. release following ingestion. Ingestion of multiples of Such a Panel C of FIG. 2 provides another example of a dose unit dose unit can provide a concentration-dose PK profile Such having a modified concentration-time PK profile. As in Panel that the relationship between number of dose units ingested A, the Solid line represents drug concentration over time in 45 and PK parameter value is linear with a positive slope, which blood or plasma following ingestion of prodrug without is similar to, for example, a dose proportional PK profile of inhibitor, while the dashed line represents drug concentration increasing amounts of prodrug alone. Panel A of FIG. 3 following ingestion of the same amount of prodrug with depicts such a profile. Dose units that provide a concentra inhibitor. In this example, the dose unit provides a PK profile tion-dose PK profile having Such an undetectable change in having a delayed drug Tmax (i.e., decreased (1/drug Tmax) 50 drug Cmax in vivo compared to the profile of prodrug alone relative to ingestion of the same dose of prodrug without can find use in thwarting enzyme conversion of prodrug from inhibitor. a dose unit that has sufficient inhibitor to reduce or prevent in Dose units that provide for a modified PK profile (e.g., a vitro cleavage of the enzyme-cleavable prodrug by its respec decreased drug Cmax and/or delayed drug Tmax as compared tive enzyme. to, a PK profile of drug or a PK profile of prodrug without 55 Panel B in FIG.3 represents a concentration-dose PK pro inhibitor), find use in tailoring of drug dose according to a file such that the relationship between the number of dose patient’s needs (e.g., through selection of a particular dose units ingested and a PK parameter value is linear with positive unit and/or selection of a dosage regimen), reduction of side slope, where the profile exhibits a reduced slope relative to effects, and/or improvement in patient compliance (as com panel A. Such a dose unit provides a profile having a pared to side effects or patient compliance associated with 60 decreased PK parameter value (e.g., drug Cmax) relative to a drug or with prodrug without inhibitor). As used herein, reference PK parameter value exhibiting dose proportional “patient compliance' refers to whether a patient follows the ity. direction of a clinician (e.g., a physician) including ingestion Concentration-dose PK profiles following ingestion of of a dose that is neither significantly above nor significantly multiples of a dose unit can be non-linear. Panel C in FIG. 3 below that prescribed. Such dose units also reduce the risk of 65 represents an example of a non-linear, biphasic concentra misuse, abuse or overdose by a patient as compared to Such tion-dose PK profile. In this example, the biphasic concen risk(s) associated with drug or prodrug without inhibitor. For tration-dose PK profile contains a first phase over which the US 8,685,916 B2 95 96 concentration-dose PK profile has a positive rise, and then a composition of inhibitor and drug provide for a modified PK second phase over which the relationship between number of profile by increasing a PK parameter. Inhibitor can also be dose units ingested and a PK parameter value (e.g., drug combined with a prodrug that is Susceptible to degradation by Cmax) is relatively flat (substantially linear with Zero slope). a GI enzyme and has a site of action outside the GI tract. In For Such a dose unit, for example, drug Cmax can be this composition, the inhibitor protects ingested prodrug in increased for a selected number of dose units (e.g. 2, 3, or 4 the GI tract prior to its distribution outside the GI tract and dose units). However, ingestion of additional dose units does cleavage at a desired site of action. not provide for a significant increase in drug Cmax. Methods Used to Define Relative Amounts of Prodrug and Panel D in FIG. 3 represents another example of a non Inhibitor in a Dose Unit linear, biphasic concentration-dose PK profile. In this 10 Dose units that provide for a desired PK profile, such as a example, the biphasic concentration-dose PK profile is char desired concentration-time PK profile and/or a desired con acterized by a first phase over which the concentration-dose centration-dose PK profile, can be made by combining a PK profile has a positive rise and a second phase over which prodrug and an inhibitor in a dose unit in relative amounts the relationship between number of dose units ingested and a effective to provide for release of drug that provides for a PK parameter value (e.g., drug Cmax) declines. Dose units 15 desired drug PK profile following ingestion by a patient. that provide this concentration-dose PK profile provide for an Prodrugs can be selected as suitable for use in a dose unit increase in drug Cmax for a selected number of ingested dose by determining the GI enzyme-mediated drug release com units (e.g., 2, 3, or 4 dose units). However, ingestion of further petency of the prodrug. This can be accomplished in vitro, in additional dose units does not provide for a significant vivo or ex vivo. increase in drug Cmax and instead provides for decreased In vitro assays can be conducted by combining a prodrug drug Cmax. with a GI enzyme (e.g., trypsin) in a reaction mixture. The GI Panel E in FIG.3 represents a concentration-dose PK pro enzyme can be provided in the reaction mixture in an amount file in which the relationship between the number of dose Sufficient to catalyze cleavage of the prodrug. Assays are units ingested and a PK parameter (e.g., drug Cmax) is linear conducted under Suitable conditions, and optionally may be with zero slope. Such dose units do not provide for a signifi 25 under conditions that mimic those found in a GI tract of a cant increase or decrease in drug Cmax with ingestion of Subject, e.g., human. "Prodrug conversion” refers to release multiples of dose units. of drug from prodrug. Prodrug conversion can be assessed by Panel F in FIG.3 represents a concentration-dose PK pro detecting a level of a product of prodrug conversion (e.g., file in which the relationship between number of dose units released drug) and/or by detecting a level of prodrug that is ingested and a PK parameter value (e.g., drug Cmax) is linear 30 maintained in the presence of the GI enzyme. Prodrug con with a negative slope. Thus drug Cmax decreases as the version can also be assessed by detecting the rate at which a number of dose units ingested increases. product of prodrug conversion occurs or the rate at which Dose units that provide for concentration-dose PK profiles prodrug disappears. An increase in released drug, or a when multiples of a dose unit are ingested find use intailoring decrease in prodrug, indicate prodrug conversion has of a dosage regimen to provide a therapeutic level of released 35 occurred. Prodrugs that exhibit an acceptable level of prodrug drug while reducing the risk of overdose, misuse, or abuse. conversion in the presence of the GI enzyme within an accept Such reduction in risk can be compared to a reference, e.g., to able period of time are suitable for use in a dose unit in administration of drug alone or prodrug alone. In one embodi combination with an inhibitor of the GI enzyme that is shown ment, risk is reduced compared to administration of a drug or to mediate prodrug conversion. prodrug that provides a proportional concentration-dose PK 40 In vivo assays can assess the Suitability of a prodrug for use profile. A dose unit that provides for a concentration-dose PK in a dose unit by 15' administration of the prodrug to an profile can reduce the risk of patient overdose through inad animal (e.g., a human or non-human animal, e.g., rat, dog. Vertent ingestion of dose units above a prescribed dosage. pig, etc.). Such administration can be enteral (e.g., oral Such a dose unit can reduce the risk of patient misuse (e.g., administration). Prodrug conversion can be detected by, for through self-medication). Such a dose unit can discourage 45 example, detecting a product of prodrug conversion (e.g., abuse through deliberate ingestion of multiple dose units. For released drug or a metabolite of released drug) or detecting example, a dose unit that provides for a biphasic concentra prodrug in blood or plasma of the animal at a desired time tion-dose PK profile can allow for an increase in drug release point(s) following administration. for a limited number of dose units ingested, after which an Ex vivo assays, such as a gut loop or inverted gut loop increase in drug release with ingestion of more dose units is 50 assay, can assess the Suitability of a prodrug for use in a dose not realized. In another example, a dose unit that provides for unitby, for example, administration of the prodrug to a ligated a concentration-dose PK profile of Zero slope can allow for section of the intestine of an animal. Prodrug conversion can retention of a similar drug release profile regardless of the be detected by, for example, detecting a product of prodrug number of dose units ingested. conversion (e.g., released drug or a metabolite of released Ingestion of multiples of a dose unit can provide for adjust 55 drug) or detecting prodrug in the ligated gut loop of the ment of a PK parameter value relative to that of ingestion of animal at a desired time point(s) following administration. multiples of the same dose (either as drug alone or as a Inhibitors are generally selected based on, for example, prodrug) in the absence of inhibitor Such that, for example, activity in interacting with the GI enzyme(s) that mediate ingestion of a selected number (e.g., 2, 3, 4 or more) of a release of drug from a prodrug with which the inhibitor is to single dose unit provides for a decrease in a PK parameter 60 be co-dosed. Such assays can be conducted in the presence of value compared to ingestion of the same number of doses in enzyme either with or without prodrug. Inhibitors can also be the absence of inhibitor. selected according to properties such as half-life in the GI Pharmaceutical compositions include those having an system, potency, avidity, affinity, molecular size and/or inhibitor to provide for protection of a therapeutic compound enzyme inhibition profile (e.g., steepness of inhibition curve from degradation in the GI tract. Inhibitor can be combined 65 in an enzyme activity assay, inhibition initiation rate). Inhibi with a drug (i.e., not a prodrug) to provide for protection of the tors for use in prodrug-inhibitor combinations can be selected drug from degradation in the GI system. In this example, the through use of in vitro, in vivo and/or ex vivo assays. US 8,685,916 B2 97 98 One embodiment is a method for identifying a prodrug and bined dose (i.e., in the same formulation). Prodrug conversion a GI enzyme inhibitor suitable for formulation in a dose unit can be detected by, for example, detecting a product of pro wherein the method comprises combining a prodrug (e.g., a drug conversion (e.g., released drug or drug metabolite) or opioid prodrug), a GI enzyme inhibitor (e.g., a trypsin inhibi detecting prodrug in blood or plasma of the animal at a tor), and a GI enzyme (e.g., trypsin) in a reaction mixture and 5 desired time point(s) following administration. Combina detecting prodrug conversion. Such a combination is tested tions of prodrug and inhibitor can be identified that provide for an interaction between the prodrug, inhibitor and enzyme, for a prodrug conversion level that yields a desired PK profile i.e., tested to determinehow the inhibitor will interact with the as compared to, for example, prodrug without inhibitor. enzyme that mediates enzymatically-controlled release of the Combinations of relative amounts of prodrug and inhibitor drug from the prodrug. In one embodiment, a decrease in 10 prodrug conversion in the presence of the GI enzyme inhibi that provide for a desired PK profile can be identified by tor as compared to prodrug conversion in the absence of the dosing animals with a fixed amount of prodrug and increasing GI enzyme inhibitor indicates the prodrug and GI enzyme amounts of inhibitor, or with a fixed amount of inhibitor and inhibitor are suitable for formulation in a dose unit. Such a increasing amounts of prodrug. One or more PK parameters method can be an in vitro assay. 15 can then be assessed, e.g., drug Cmax, drug Tmax, and drug One embodiment is a method for identifying a prodrug and exposure. Relative amounts of prodrug and inhibitor that a GI enzyme inhibitor suitable for formulation in a dose unit provide for a desired PK profile are identified as amounts of wherein the method comprises administering to an animal a prodrug and inhibitor for use in a dose unit. The PK profile of prodrug and a GI enzyme inhibitor and detecting prodrug the prodrug and inhibitor combination can be, for example, conversion. In one embodiment, a decrease in prodrug con characterized by a decreased PK parameter value relative to version in the presence of the GI enzyme inhibitor as com prodrug without inhibitor. A decrease in the PK parameter pared to prodrug conversion in the absence of the GI enzyme value of an inhibitor-to-prodrug combination (e.g., a decrease inhibitor indicates the prodrug and GI enzyme inhibitor are in drug Cmax, a decrease in 1/drug Tmax (i.e., a delay in drug suitable for formulation in a dose unit. Such a method can be Tmax) or a decrease in drug exposure) relative to a corre an in vivo assay; for example, the prodrug and GI enzyme 25 sponding PK parameter value following administration of inhibitor can be administered orally. Such a method can also prodrug without inhibitor can be indicative of an inhibitor-to be an ex vivo assay; for example, the prodrug and GI enzyme prodrug combination that can provide a desired PK profile. inhibitor can be administered orally or to a tissue. Such as an Assays can be conducted with different relative amounts of intestine, that is at least temporarily exposed. Detection can inhibitor and prodrug. occur in the blood or plasma or respective tissue. As used 30 In vivo assays can be used to identify combinations of herein, tissue refers to the tissue itself and can also refer to contents within the tissue. prodrug and inhibitor that provide for dose units that provide One embodiment is a method for identifying a prodrug and for a desired concentration-dose PK profile following inges a GI enzyme inhibitor suitable for formulation in a dose unit tion of multiples of the dose unit (e.g., at least 2, at least 3, at wherein the method comprises administering a prodrug and a 35 least 4 or more). Ex vivo assays can be conducted by direct gastrointestinal (GI) enzyme inhibitor to an animal tissue that administration of prodrug and inhibitor into a tissue and/or its has removed from an animal and detecting prodrug conver contents of an animal. Such as the intestine, including by Sion. In one embodiment, a decrease in prodrug conversion in introduction by injection into the lumen of a ligated intestine the presence of the GI enzyme inhibitor as compared to pro (e.g., a gut loop, or intestinal loop, assay, or an inverted gut drug conversion in the absence of the GI enzyme inhibitor 40 assay). An ex vivo assay can also be conducted by excising a indicates the prodrug and GI enzyme inhibitor are suitable for tissue and/or its contents from an animal and introducing formulation in a dose unit. prodrug and inhibitor into Such tissues and/or contents. In vitro assays can be conducted by combining a prodrug, For example, a dose of prodrug that is desired for a single an inhibitor and a GI enzyme in a reaction mixture. The GI dose unit is selected (e.g., an amount that provides an effica enzyme can be provided in the reaction mixture in an amount 45 cious plasma drug level). A multiple of single dose units for Sufficient to catalyze cleavage of the prodrug, and assays which a relationship between that multiple and a PK param conducted under Suitable conditions, optionally under condi eter to be tested is then selected. For example, if a concentra tions that mimic those found in a GI tract of a Subject, e.g., tion-dose PK profile is to be designed for ingestion of 2, 3, 4, human. Prodrug conversion can be assessed by detecting a 5, 6, 7, 8, 9 or 10 dose units, then the amount of prodrug level of a product of prodrug conversion (e.g., released drug) 50 equivalent to ingestion of that same number of dose units is and/or by detecting a level of prodrug maintained in the determined (referred to as the “high dose”). The multiple of presence of the GI enzyme. Prodrug conversion can also be dose units can be selected based on the number of ingested assessed by detecting the rate at which a product of prodrug pills at which drug Cmax is modified relative to ingestion of conversion occurs or the rate at which prodrug disappears. the single dose unit. If, for example, the profile is to provide Prodrug conversion that is modified in the presence of inhibi 55 for abuse deterrence, then a multiple of 10 can be selected, for tor as compared to a level of prodrug conversion in the example. A variety of different inhibitors (e.g., from a panel absence of inhibitor indicates the inhibitor is suitable for of inhibitors) can be tested using different relative amounts of attenuation of prodrug conversion and for use in a dose unit. inhibitor and prodrug. Assays can be used to identify Suitable Reaction mixtures having a fixed amount of prodrug and combination(s) of inhibitor and prodrug to obtain a single increasing amounts of inhibitor, or a fixed amount of inhibitor 60 dose unit that is therapeutically effective, wherein such a and increasing amounts of prodrug, can be used to identify combination, when ingested as a multiple of dose units, pro relative amounts of prodrug and inhibitor which provide for a vides a modified PK parameter compared to ingestion of the desired modification of prodrug conversion. same multiple of drug or prodrug alone (wherein a single dose In vivo assays can assess combinations of prodrugs and of either drug or prodrug alone releases into blood or plasma inhibitors by co-dosing of prodrug and inhibitor to an animal. 65 the same amount of drug as is released by a single dose unit). Such co-dosing can be enteral. "Co-dosing refers to admin Increasing amounts of inhibitor are then co-dosed to ani istration of prodrug and inhibitor as separate doses or a com mals with the high dose of prodrug. The dose level of inhibitor US 8,685,916 B2 99 100 that provides a desired drug Cmax following ingestion of the frangible seal elements include a seal positioned over a con high dose of prodrug is identified and the resultant inhibitor tainer opening such that access to a dose unit within the to-prodrug ratio determined. container requires disruption of the seal (e.g., by peeling Prodrug and inhibitor are then co-dosed in amounts equiva and/or piercing the seal). Examples of frangible seal elements lent to the inhibitor-to-prodrug ratio that provided the desired include a frangible ring disposed around a container opening result at the high dose of prodrug. The PK parameter value of and in connection with a cap Such that the ring is broken upon interest (e.g., drug Cmax) is then assessed. If a desired PK opening of the cap to access the dose units in the container. parameter value results followingingestion of the single dose Dry and liquid dose units can be placed in a container (e.g., unit equivalent, then single dose units that provide for a bottle or package, e.g., a flexible bag) of a size and configu desired concentration-dose PK profile are identified. For 10 ration adapted to maintain stability of dose units over a period example, where a Zero dose linear profile is desired, the drug during which the dose units are dispensed into a prescription. Cmax following ingestion of a single dose unit does not For example, containers can be sized and configured to con increase significantly following ingestion of a multiple num tain 10, 20, 30, 40, 50, 60, 70, 80,90, 100 or more single dry ber of the single dose units. or liquid dose units. The containers can be sealed or reseal Methods for Manufacturing. Formulating, and Packaging 15 able. The containers can packaged in a carton (e.g., for ship Dose Units ment from a manufacturer to a pharmacy or other dispensary). Dose units of the present disclosure can be made using Such cartons can be boxes, tubes, or of other configuration, manufacturing methods available in the art and can be of a and may be made of any material (e.g., cardboard, plastic, and variety of forms suitable for enteral (including oral, buccal the like). The packaging system and/or containers disposed and Sublingual) administration, for example as a tablet, cap therein can have one or more affixed labels (e.g., to provide Sule, thin film, powder, Suspension, Solution, syrup, disper information Such as lot number, dose unit type, manufacturer, sion or emulsion. The dose unit can contain components and the like). conventional in pharmaceutical preparations, e.g. one or The container can include a moisture barrier and/or light more carriers, binders, lubricants, excipients (e.g., to impart barrier, e.g., to facilitate maintenance of stability of the active controlled release characteristics), pH modifiers, flavoring 25 ingredients in the dose units contained therein. Where the agents (e.g., Sweeteners), bulking agents, coloring agents or dose unit is a dry dose unit, the container can include a further active agents. Dose units of the present disclosure can desiccant pack which is disposed within the container. The include can include an enteric coating or other component(s) container can be adapted to contain a single dose unit or to facilitate protection from stomach acid, where desired. multiples of a dose unit. The container can include a dispens Dose units can be of any suitable size or shape. The dose 30 ing control mechanism, such as a lock out mechanism that unit can be of any shape Suitable for enteral administration, facilitates maintenance of dosing regimen. e.g., ellipsoid, lenticular, circular, rectangular, cylindrical, The dose units can be provided in solid or semi-solid form, and the like. and can be a dry dose unit. "Dry dose unit refers to a dose Dose units provided as dry dose units can have a total unit that is in other than in a completely liquid form. weight of from about 1 microgram to about 1 gram, and can be 35 Examples of dry dose units include, for example, tablets, from about 5 micrograms to 1.5 grams, from about 50 micro capsules (e.g., Solid capsules, capsules containing liquid). grams to 1 gram, from about 100 micrograms to 1 gram, from thin film, microparticles, granules, powder and the like. Dose 50 micrograms to 750 milligrams, and may be from about 1 units can be provided as liquid dose units, where the dose microgram to 2 grams. units can be provided as single or multiple doses of a formu Dose units can comprise components in any relative 40 lation containing prodrug and inhibitor in liquid form. Single amounts. For example, dose units can be from about 0.1% to doses of a dry or liquid dose unit can be disposed within a 99% by weight of active ingredients (i.e., prodrug and inhibi sealed container, and sealed containers optionally provided in tor) pertotal weight of dose unit (0.1% to 99% total combined a packaging system, e.g., to provide for a prescribed number weight of prodrug and inhibitor pertotal weight of single dose of doses, to provide for shipment of dose units, and the like. unit). In some embodiments, dose units can be from 10% to 45 Dose units can be formulated Such that the prodrug and 50%, from 20% to 40%, or about 30% by weight of active inhibitor are present in the same carrier, e.g., Solubilized or ingredients per total weight dose unit. Suspended within the same matrix. Alternatively, dose units Dose units can be provided in a variety of different forms can be composed of two or more portions, where the prodrug and optionally provided in a manner Suitable for storage. For and inhibitor can be provided in the same or different por example, dose units can be disposed within a container Suit 50 tions, and can be provided in adjacent or non-adjacent por able for containing a pharmaceutical composition. The con tions. tainer can be, for example, a bottle (e.g., with a closure device, Dose units can be provided in a container in which they are Such as a cap), a blister pack (e.g., which can provide for disposed, and may be provided as part of a packaging system enclosure of one or more dose units per blister), a vial, flexible (optionally with instructions for use). For example, dose units packaging (e.g., sealed Mylar or plastic bags), an ampule (for 55 containing different amounts of prodrug can be provided in single dose units in Solution), a dropper, thin film, a tube and separate containers, which containers can be disposed within the like. a larger container (e.g., to facilitate protection of dose units Containers can include a cap (e.g., Screw cap) that is for shipment). For example, one or more dose units as removably connected to the container over an opening described herein can be provided in separate containers, through which the dose units disposed within the container 60 where dose units of different composition are provided in can be accessed. separate containers, and the separate containers disposed Containers can include a seal which can serve as a tamper within package for dispensing. evident and/or tamper-resistant element, which seal is dis In another example, dose units can be provided in a double rupted upon access to a dose unit disposed within the con chambered dispenser where a first chamber contains a pro tainer. Such seal elements can be, for example, a frangible 65 drug formulation and a second chamber contains an inhibitor element that is broken or otherwise modified upon access to a formulation. The dispenser can be adapted to provide for dose unit disposed within the container. Examples of Such mixing of a prodrug formulation and an inhibitor formulation US 8,685,916 B2 101 102 prior to ingestion. For example, the two chambers of the The present disclosure provides methods to reduce misuse dispenser can be separated by a removable wall (e.g., fran and abuse of a drug, as well as to reduce risk of overdose, that gible wall) that is broken or removed prior to administration can accompany ingestion of multiples of doses of a drug, e.g., to allow mixing of the formulations of the two chambers. The ingested at the same time. Such methods generally involve first and second chambers can terminate into a dispensing combining in a dose unit a prodrug and an inhibitor of a GI outlet, optionally through a common chamber. The formula enzyme that mediates release of drug from the prodrug, where tions can be provided in dry or liquid form, or a combination the inhibitor is present in the dose unit in an amount effective thereof. For example, the formulation in the first chamber can to attenuate release of drug from the prodrug, e.g., following be liquid and the formulation in the second chamber can be ingestion of multiples of dose units by a patient. Such meth 10 ods provide for a modified concentration-dose PK profile dry, both can be dry, or both can be liquid. while providing therapeutically effective levels from a single Dose units that provide for controlled release of prodrug, of dose unit, as directed by the prescribing clinician. Such meth inhibitor, or of both prodrug and inhibitor are contemplated ods can provide for, for example, reduction of risks that can by the present disclosure, where “controlled release' refers to accompany misuse and/or abuse of a prodrug, particularly release of one or both of prodrug and inhibitor from the dose 15 where conversion of the prodrug provides for release of a unit over a selected period of time and/or in a pre-selected narcotic or other drug of abuse (e.g., opioid). For example, a. when the prodrug provides for release of a drug of abuse, dose Methods of Use of Dose Units units can provide for reduction of reward that can follow Dose units are advantageous because they find use in meth ingestion of multiples of dose units of a drug of abuse. ods to reduce side effects and/or improve tolerability of drugs Dose units can provide clinicians with enhanced flexibility to patients in need thereof by, for example, limiting a PK in prescribing drug. For example, a clinician can prescribe a parameter as disclosed herein. The present disclosure thus dosage regimen involving different dose strengths, which can provides methods to reduce side effects by administering a involve two or more different dose units of prodrug and dose unit of the present disclosure to a patient in need so as to inhibitor having different relative amounts of prodrug, differ provide for a reduction of side effects as compared to those 25 ent amounts of inhibitor, or different amounts of both prodrug associated with administration of drug and/or as compared to and inhibitor. Such different strength dose units can provide administration of prodrug without inhibitor. The present dis for delivery of drug according to different PK parameters closure also provides methods to improve tolerability of (e.g., drug exposure, drug Cmax, and the like as described drugs by administering a dose unit of the present disclosure to herein). For example, a first dose unit can provide for delivery a patient in need so as to provide for improvement in toler 30 of a first dose of drug following ingestion, and a second dose ability as compared to administration of drug and/or as com unit can provide for delivery of a second dose of drug follow pared to administration of prodrug without inhibitor. ing ingestion. The first and second prodrug doses of the dose Dose units find use in methods for increasing patient com units can be different strengths, e.g., the second dose can be pliance of a patient with a therapy prescribed by a clinician, greater than the first dose. A clinician can thus prescribe a where such methods involve directing administration of a 35 collection of two or more, or three or more dose units of dose unit described hereinto a patient in need of therapy so as different strengths, which can be accompanied by instruc to provide for increased patient compliance as compared to a tions to facilitate a degree of self-medication, e.g., to increase therapy involving administration of drug and/or as compared delivery of an opioid drug according to a patient’s needs to to administrations of prodrug without inhibitor. Such meth treat pain. ods can help increase the likelihood that a clinician-specified 40 Thwarting Tampering by Trypsin Mediated Release of therapy occurs as prescribed. Opioid from Prodrugs Dose units can provide for enhanced patient compliance The disclosure provides for a composition comprising a and clinician control. For example, by limiting a PK param compound disclosed herein and a trypsin inhibitor that eter (e.g., Such as drug Cmax or drug exposure) when mul reduces drug abuse potential. A trypsin inhibitor can thwart tiples (e.g., two or more, three or more, or four or more) dose 45 the ability of a user to apply trypsin to effect the release of an units are ingested, a patient requiring a higher dose of drug opioid from the opioid prodrug in vitro. For example, if an must seek the assistance of a clinician. The dose units can abuser attempts to incubate trypsin with a composition of the provide for control of the degree to which a patient can readily embodiments that includes an opioid prodrug and a trypsin “self-medicate', and further can provide for the patient to inhibitor, the trypsin inhibitor can reduce the action of the adjust dose to a dose within a permissible range. Dose units 50 added trypsin, thereby thwarting attempts to release the can provide for reduced side effects, by for example, provid opioid for purposes of abuse. ing for delivery of drug at an efficacious dose but with a modified PK profile over a period of treatment, e.g., as EXAMPLES defined by a decreased PK parameter (e.g., decreased drug Cmax, decreased drug exposure). 55 The following examples are put forth so as to provide those Dose units find use in methods to reduce the risk of unin of ordinary skill in the art with a complete disclosure and tended overdose of drug that can follow ingestion of multiple description of how to make and use the embodiments, and are doses taken at the same time or over a short period of time. not intended to limit the scope of what the inventors regard as Such methods of the present disclosure can provide for reduc their invention nor are they intended to represent that the tion of risk of unintended overdose as compared to risk of 60 experiments below are all or the only experiments performed. unintended overdose of drug and/or as compared to risk of Efforts have been made to ensure accuracy with respect to unintended overdose of prodrug without inhibitor. Such numbers used (e.g. amounts, temperature, etc.) but some methods involve directing administration of a dosage experimental errors and deviations should be accounted for. described herein to a patient in need of drug released by Unless indicated otherwise, parts are parts by weight, conversion of the prodrug. Such methods can help avoid 65 molecular weight is weight average molecular weight, tem unintended overdosing due to intentional or unintentional perature is in degrees Celsius, and pressure is at or near misuse of the dose unit. atmospheric. Standard abbreviations may be used. US 8,685,916 B2 103 104 Synthesis of Ketone-modified Opioid Prodrugs (330 mg, 5% on carbon) was added. The mixture was shaken in a Parr hydrogenator flask (50 psi H) for 4 h. The mixture Example 1 was then filtered through a celite pad, and the filtrate was concentrated to give Compound B (1.08 g, yield exceeded Synthesis of oxycodone 6-(N-methyl-N-(2-amino) quantitative). Compound B was used without further purifi ethylcarbamate (Compound KC-19) cation. Preparation of Compound C Compound B (500mg, 1.82 mmol) and NEt (0.4 mL, 2.74 mmol) were mixed together in dichloromethane (4 mL). The 10 mixture was added to a pre-chilled 0°C. solution of phosgene (5.5 mL, 0.5M in toluene). The reaction mixture was stirred at 0°C. for 1 h, followed by dilution with ether (20 mL) and filtration through filter paper. The filtrate was concentrated 15 and passed through a short silica gel column (10 cmx3 cm). and eluted with 3/1 hexanes/EtOAc. The fractions were con centrated to give N,N-Bis(tert-butyl) N'-2-(chlorocarbonyl (methyl)amino)ethylcarbamate (Compound C) as a colorless solid in quantitative yield (615 mg, 1.82 mmol). MS: (m/z) calc: 336.1, observed (M+Na") 359.8. Synthesis of Oxycodone 6-(N-methyl-N-(2-amino) ethylcarbamate (Compound KC-19)

25 1. KHMDS:

30

phosgene s O Her C r --~. N& K 2. TFADCM

40 N1 OH

45

-O O O--~-Ns

KC-19 50 Preparation of Compound A Oxycodone free base (6.5g, 20.6 mmol) was dissolved in 2-(Aminoethyl)-methyl-carbamic acid benzyl ester (2.0 g, dry, degassed tetrahydrofuran (120 mL), and the mixture was 9.6 mmol) was dissolved in dichloroethene (DCE) (20 mL) at cooled to -10°C. using a dry ice/acetone bath. Potassium room temperature. Triethyl amine (NEt) (1.40 mL, 11.5 55 bis(trimethylsilyl)amide (KHMDS) (103.0 mL, 51.6 mmol, mmol) was added, followed by di-tert-butyl dicarbonate 0.5 M in toluene) was added via cannula. The mixture was (BOCO) (10.5 g, 48 mmol) and dimethylaminopyridine stirred under N at below -5°C. for 30 min. N,N-Bis(tert (DMAP) (120 mg). The reaction mixture was stirred at room butyl) N'-2-(chlorocarbonyl(methyl)amino)ethylcarbamate temperature under nitrogen (N) for 2 hand then heated at 60° (8.0g, 23.7 mmol), (Compound C) in THF (30 mL) was then C. for 16 h. The reaction mixture was then concentrated. The 60 added via cannula over 15 min. The mixture was stirred at -5° residue was purified by Silica gel chromatography, using 4/1 C. for 30 min. Another portion of carbamoyl chloride (4.0 g, hexanes/EtOAc, to give Compound A in 86% yield (3.4g, 8.3 11.9 mmol) in THF (10 mL) was added. The reaction was mmol). MS: (m/z) calc: 408.2, observed (M+Na") 431.9. stirred at room temperature for 2 h. Sodium bicarbonate (10 Preparation of Compound B mL, sat. aq.) was added. The mixture was concentrated under Compound A (1.3 g, 3.18 mmol) was dissolved in metha 65 vacuum to half of its initial volume. EtOAc (50 mL) was nol/EtOAc (10 mL/3 mL respectively). The mixture was added, and layers were separated. The organic phase was degassed and saturated with N. Palladium on carbon (Pd/C) further washed with water (3x20 mL) and brine (40 mL), and US 8,685,916 B2 105 106 then was concentrated. The residue was purified by silica gel Example 2 chromatography, using DCM/MeOH (gradient 100/1 to 100/ 15) to afford a white foam in 55% yield (7.0g, 13.4 mmol). This material was dissolved in a 1:1 mixture of DCM/trifluo Synthesis of N-1-2-(oxycodone-6-enol-carbonyl roacetic acid (TFA) (20 mL/20 mL) at room temperature and 5 methyl-amino)-ethylamine-L-arginine-malonate stirred for 1 h. The solution was then concentrated under (Compound KC-3) also named: N-(S)-4-guani vacuum to afford a TFA salt of oxycodone 6-(N-methyl-N- dino-1-2-(methyl-(5R.9R,13S, 14S)-4,5a-epoxy-6, (2-amino)ethylcarbamate (Compound KC-19) as a thick oil 7-didehydro-14-hydroxy-3-methoxy-17-methylmor (7.3 g, 11.4 mmol. 99% purity). MS: (m/z) calc: 415.2. phinan-6-oxycarbonyl-amino)-ethylcarbamoyl observed (M+H") 416.5. butyl-malonate

CFC(O)OEt He 2 ACN, water

N NY YPbf NH2 Boc-Arg(Pbf)-OH, HATU, DIEA \ se b OC N1\- r H ch/N \ NH O F close H N. N N N Y n Pbf O NY YPbf NH2 NH2 -u X O O H BOP, DIEA Sr- N 1st, Cbz O --- H I pric MeOH N NY YPbf NH2 O O H NN 1-N-N H 1sH O O

OC, KHMDS, 4-NPCF US 8,685,916 B2 107 108 -continued N Y YPbf NH2 O O - O us,uk H

1. TFA, 5% m-cresol 2. Lyo with 0.1N HCI

Preparation of Compound D (CHNO+H, calc: 209.3). Compound F was used A solution of N-methylethylenediamine (27.0 g, 364 30 directly in the next reaction without purification as a DMF mmol) and ethyl trifluoroacetate (96.6 mL, 812 mmol) in a Solution. mixture of ACN (350 mL) and water (7.8 mL, 436 mmol) was Preparation of Compound G refluxed with stirring overnight. Solvents were evaporated A solution of Boc-Arg(Pbf)-OH (16.0 g, -30.4 mmol), under vacuum. The residue was re-evaporated with i-PrCH 35 Compound F hydrochloride (8.2g, 33.4 mmol), and DIEA (3x100 mL), followed by heat-cool crystallization from (16.9 mL. 97.2 mmol) in DMF (150 mL) was cooled in an ice DCM (500 mL). Formed crystals were filtered, washed with bath followed by the addition of a solution of HATU (13.8 g. DCM and dried under vacuum to provide Compound D (88.3 36.4 mmol) dropwise over 20 min. The temperature of the g, 85%) as white solid powder. reaction mixture was raised to ambient temperature, and stir Preparation of Compound E 40 ring was continued for an additional 1 h. The reaction mixture A solution of Compound D (88.2g, 311 mmol) and DIEA was diluted with EtOAc (1 L) and extracted with water (54.1 mL, 311 mmol) in THF (350 mL) was cooled in an ice (3x200 mL) and brine (200 mL). The organic layer was dried bath, followed by the addition of a solution of N-(benzyloxy over MgSO and evaporated to provide Compound G (24.4g, carbonyl)succinimide (76.6 g. 307 mmol) in THF (150 mL) yield exceeded quantitative) as a yellowish oil. LC-MS dropwise over the period of 20 min. The temperature of the 45 M+H 717.4 (CHNOS+H, calc: 717.9). Compound G reaction mixture was raised to ambient temperature and stir was used directly in the next reaction without purification as ring was continued for an additional 30 min. Solvents were a dioxane Solution. then evaporated and the resulting residue was dissolved in Preparation of Compound H EtOAc (600 mL). The organic layer was extracted with 5% Compound G (24.4g, ~30.4 mmol) was dissolved in diox aq. NaHCO (2x150 mL) and brine (150 mL). The organic 50 ane (150 mL) and treated with 4 NHCl/dioxane (150 mL, 600 layer was evaporated to provide Compound E as yellowish mmol) at ambient temperature for 1 h. The solvent was then oil. LC-MS M+H 305.1 (CHFNO+H, calc: 305.3). evaporated. The residue was suspended in i-PrCH (100 mL), Compound E was used directly in the next reaction without and the mixture was evaporated (procedure was repeated purification as a MeOH solution. twice). The residue was then dried under vacuum to provide Preparation of Compound F 55 Compound H (21.1 g, yield exceeded quantitative) as a yel To a solution of Compound E (-311 mmol) in MeCH (1.2 lowish solid. LC-MSM-I-H 617.5 (CHNOS+H, calc: L) was added a solution of LiOH (14.9 g, 622 mmol) in water 617.8). Compound H was used directly in the next reaction (120 mL). The reaction mixture was stirred at ambient tem without purification as a DMF solution. perature for 3 h. Solvents were evaporated to 75% of the Preparation of Compound I initial volume followed by dilution with water (400 mL). The 60 A solution of Compound H (21.1 g, ~30.4 mmol), mono solution was extracted with EtOAc (2x300 mL). The organic tert-butyl malonate (5.9 mL, 36.7 mmol), BOP (16.2g, 36.7 layer was washed with brine (200 mL), dried over MgSO and mmol) and DIEA (14.9 mL, 83.5 mmol) in DMF (100 mL) evaporated under vacuum. The residue was dissolved in ether was maintained at ambient temperature for 1 h. The reaction (300 mL) and treated with 2 NHCl/ether (200 mL). Formed mixture was diluted with EtOAc (1 L) and extracted with precipitate was filtrated, washed with ether and dried under 65 water (500 mL), 5% aq. NaHCO, (500 mL), water (3x500 vacuum to provide the hydrochloric salt of Compound F (67.8 mL) and brine (500 mL). The organic layer was dried over g, 89%) as a white solid. LC-MS M+H 209.0 MgSO filtered, and then evaporated to provide Compound I US 8,685,916 B2 109 110 (24.5 g, 97%) as a yellowish amorphous solid. LC-MS Synthesis of N-1-2-(oxycodone-6-enol-carbonyl M+H 759.6 (CHNOS+H, calc: 759.9). Compound I methyl-amino)-ethylamine-L-arginine-malonate was used without further purification. (Compound KC-3) Preparation of Compound J Compound I (12.3 g 16.7 mmol) was dissolved in metha nol (100 mL) followed by the addition of a Pd/C (5% wt, 2.0 Compound K (13.6 g., 14.1 mmol) was dissolved in a mix g) Suspension in water (2 mL). The reaction mixture was ture of 5% m-cresol/TFA (100 mL). The reaction mixture was Subjected to hydrogenation (Parr apparatus, 70 psi H) at maintained at ambient temperature for 1 h, followed by dilu ambient temperature for 1 h. The catalyst was then filtered tion with ethyl ether (1 L). The formed precipitate was fil and washed with methanol. The filtrate was evaporated under 10 tered, washed with ether and hexane, and dried under vacuum vacuum to provide Compound J (10.0 g, 99%) as a colorless to provide a TFA salt of Compound KC-3 (11.4g, 81%) as an amorphous solid. LC-MSM-I-H 625.5 (CHNO,S+H, off-white solid. LC-MSM-H 658.6 (CHN,O+H, calc: calc: 625.8). Compound J was used without further purifica 658.7). tion. The TFA salt of crude Compound KC-3 (11.4 g, 11.4 Preparation of Oxycodone Free Base 15 mmol) was dissolved in water (50 mL). The obtained solution Oxycodone hydrochloride (10.0 g, 28.5 mmol) was dis was subjected to HPLC purification. Nanosyn-Pack YMC solved in chloroform (150 mL) and washed with 5% aq. NaHCO (50 mL). The organic layer was dried over MgSO GEL-ODSA (100-10) C-18 column (75x500 mm); flow rate: and evaporated. The residue was dried under vacuum over 250 mL/min: injection volume 50 mL. mobile phase A: 100% night to provide oxycodone free base (8.3g, 93%) as a white water, 0.1% TFA: mobile phase B: 100% ACN, 0.1% TFA: solid. isocratic elution at 0% B in 4 min, gradient elution from 0% Preparation of Compound K to 10% B in 20 min, isocratic elution at 10% B in 30 min, A solution of oxycodone free base (6.6 g. 21.0 mmol) in gradient elution from 10% B to 30% B in 41 min; detection at 254 nm. Fractions containing Compound KC-3 were com THF (400 mL) was cooled to -20°C., followed by addition of bined and concentrated under vacuum. The TFA counterion a 0.5 M solution of KHMDS in toluene (46.3 mL, 23.1 25 mmol). The obtained solution was then added to a solution of of the latter was replaced with an HCl counterion via lyo 4-nitro-phenyl chloroformate (4.3 g 21.0 mmol) in THF (100 philization using 0.1N HCl to provide an HCl salt of Com mL) dropwise over the period of 20 min at -20° C. The pound KC-3 (4.2 g, 41% yield) as a white solid. LC-MS reaction was maintained at -20° C. for an additional 1 h, M+H 658.6 (CHNO+H, calc: 658.7). followed by addition of a solution of Compound J (10.0 g, 30 16.1 mmol) in THF (200 mL) at -20°C. The reaction mixture Example 3 was allowed to warm to ambient temperature and stirred overnight. Solvents were evaporated under vacuum. The resulting residue was dissolved in EtOAc (20 mL) and pre Synthesis of N-(oxycodone-6-enol-carbonyl)piperi cipitated with ether (1 L). The formed precipitate was fil 35 dine-2-methylamine (Compound KC-11) and trated, washed with ether and dried under vacuum to provide N-(oxycodone-6-enol-carbonyl)piperidine-2-methy Compound K (13.6 g., 87%) as an off-white solid. LC-MS lamine-L-arginine-L-alanine-acetate (Compound M+H 966.9 (CH7NOS+H, calc: 966.2). KC-13)

KHMDS;

US 8,685,916 B2 111 112 -continued

HATU, DIEA

KC-11

HC Dioxane

N Boc-Ala-OH NH HATU, DIEA

1. Pbf N l N H H

HC Dioxane US 8,685,916 B2 113 114 -continued

O E N NH2 Ac2O, DIEA O Hip NH

N ls N1 Pbf H H

O O

N TFAm-Cresol -e- NH N N

O O

N N NullsN H 1s O NH

H 1. 2

KC-13

Preparation of Oxycodone Free Base (L): mL). This reaction mixture was then stirred for 30 minat-60° Oxycodone-hydrochloride (21.0 g, 59.7 mmol) was dis C., followed by addition of piperidine-2-yl-methylcarbamic solved in water (250 mL). This solution was basified with acid tert-butyl ester, also referred to herein as (R.S)-piperi saturated aqueous NaHCO (to pH 8-9) and extracted with 55 dine-2-yl-methylcarbamic acid tert-butyl ester, (5.0 g, 23.3 DCM (3x250 mL). The combined organic layer was dried mmol) in portions. The reaction was allowed to warm to over NaSO and filtered; removal of solvents under vacuum ambient temperature and then stirred for 18 h. The reaction afforded Compound L in 98% yield (18.5g, 58.8 mmol) as a was then concentrated under vacuum, and the residue diluted white solid. LC-MS M--H 316.1 (CHNO+H, calc: with EtOAc (500 mL). The mixture was then washed with 316.2). Compound L was used directly in the next reaction 60 without further purification. water (2x250 mL) and brine (250 mL). The organic layer was Preparation of Compound N separated, dried over NaSO, and filtered. Removal of sol To a solution of CompoundL (14.71 g, 46.7 mmol) in THF vents under vacuum afforded crude Compound N. Crude (250 mL) at –60° C. was added 0.5 M KHMDS solution in Compound N was purified by flash chromatography using THF (103 mL) dropwise. After stirring at-60° C. for 30 min, 65 100% EtOAc. Removal of solvent under vacuum afforded the reaction mixture was added to a solution of 4-nitrophenyl Compound N in 50% yield (6.5 g., 11.7 mmol) as a white chloroformate at –60° C. (9.41 g, 46.7 mmol) in THF (200 solid. LC-MS M+H 556.1 (CHNO+H, calc: 555.3). US 8,685,916 B2 115 116 Preparation of N-(oxycodone-6-enol-carbonyl)pip white solid. LC-MSM-I-H 935.8 (CHNOS+H, calc: eridine-2-methylamine) (KC-11) 935.5). Compound R was used directly in the next reaction without further purification. A solution of Compound N (6.5 g, 11.7 mmol) in 1,4- Preparation of Compound S dioxane (100 mL) was treated with hydrogen chloride (4.0M To a solution of Compound R (0.67 g. 0.66 mmol) and solution in 1,4-dioxane, 100 mL). After 1 h, most of the DIEA (0.37 mL, 2.1 mmol) in CHCl (50 mL) and cooled to 1,4-dioxane was removed under vacuum to -20 mL remain -0° C., was added acetic anhydride (AcO) (0.07 mL, 0.7 ing. To this solution was added EtO (-750 mL). The product mmol). The reaction mixture was stirred at ambient tempera was then precipitated as an HCl salt. The precipitate was ture for 30 min. The reaction mixture was diluted with CHCl filtered, washed with ether and dried under vacuum to afford 10 (50 mL), and washed with water (2x100 mL) and brine (50 Compound KC-11 in 97% yield (5.96 g, 11.3 mmol) as a mL). The organic layer was separated, dried over NaSO, white solid. LC-MS IM+H 456.3 (CHNO+H, calc: and filtered. Removal of solvents under vacuum afforded the 456.2). Compound KC-11 was used directly in the next reac crude CompoundS, yield exceeded quantitative, (0.65 g, 0.66 tion without further purification. mmol) as an off-white solid. LC-MS M+H 977.4 Preparation of Compound O 15 (CHNOS+H, calc: 977.5). Compound S was used To a solution of Boc-Arg(Pbf)-OH (5.94 g, 11.3 mmol), directly in the next reaction without further purification. Compound KC-11 (5.95 g, 11.3 mmol) and DIEA (8.24 mL, 47.4 mmol) in DMF (100 mL) at -0° C. was added HATU Preparation of N-(oxycodone-6-enol-carbonyl)pip (4.28 g, 11.3 mmol) in portions over 10 min. The temperature eridine-2-methylamine-L-arginine-L-alanine-acetate of the reaction mixture was raised to ambient temperature and (Compound KC-13) stirring was continued for an additional 1 h. DMF was Compound S (0.65 g, 0.66 mmol) was treated with 5% removed under vacuum, and the reaction mixture was diluted m-cresol in TFA (15 mL) for 1 h. The product was precipi with EtOAc (300 mL), washed with water (3x150 mL) and tated via addition of EtO (100 mL). The precipitate was brine (150 mL). The organic layer was separated, dried over 25 washed with EtO (2x100 mL) and dried under vacuum to NaSO, and filtered. Removal of solvents under vacuum afford crude Compound KC-13. This product was dissolved afforded crude Compound O. This compound was purified by in water (15 mL), and the solution was subjected to HPLC silica gel chromatography using CHCl and 0% to 20% purification. Nanosyn-Pack Microsorb (100-10) C-18 col MeOH. Removal of solvents under vacuum afforded Com umn (50x300mm); flow rate: 100 mL/min: injection volume pound O in 23% yield (2.5g, 2.6 mmol) as a foamy solid. 30 15 mL. mobile phase A: 100% water, 0.1% TFA; mobile LC-MSM--H964.8 (CHNOS+H, calc: 964.5). phase B: 100% ACN, 0.1%TFA: isocratic elution at 0% B in Preparation of Compound P 5 min, gradient elution from 0% to 20% B in 20 min, isocratic A solution of Compound O (2.5 g. 2.6 mmol) in 1,4- elution at 20% B in 20 min, gradient elution from 20% B to dioxane (50 mL) was treated with hydrogen chloride (4.0 M 45% B in 40 min; detection at 254 nm. Fractions containing solution in 1,4-dioxane, 50 mL). After 1 h, most of the 1,4- 35 the desired compound were combined and concentrated dioxane was removed under vacuum until ~10 mL remained. under vacuum. The residue was dissolved in ACN (-2 mL) To this solution was added EtO (-500 mL). The product and 0.1 NHCl (-8 mL), and lyophilized overnight to provide precipitated as an HCl salt. The precipitate was filtered off, the hydrochloric salt of Compound KC-13 in 90% yield (0.65 washed with ether, and dried under vacuum to afford Com g, 0.59 mmol. 93.1% purity) as a white solid. LC-MSM-H pound P in 52% yield (1.25 g, 1.33 mmol) as a white solid. 40 LC-MSM-I-H 864.6 (CHNOS+H, calc: 863.4). Com 725.8 (CHNO+H, calc: 725.4). pound P was used directly in the next reaction without further purification. Example 4 Preparation of Compound Q Synthesis of N-(oxycodone-6-enol-carbonyl)pyrroli To a solution of Boc-Ala-OH (0.13 g, 0.66 mmol), Com 45 dine-2-methylamine (Compound KC-9) pound P (0.62 g, 0.66 mmol), and DIEA (0.48 mL, 2.77 mmol) in DMF (10 mL) at 5° C., was added HATU (0.25g, 0.66 mmol) in portions over 5 min. The temperature of the reaction mixture was raised to ambient temperature, and stir ring was continued for an additional 1 h. DMF was removed 50 under vacuum. Next the reaction mixture was diluted with EtOAc (100 mL), and washed with water (3x50 mL) and brine (50 mL). The organic layer was separated, dried over NaSO, and filtered. Removal of solvents under vacuum afforded crude Compound Q, yield exceeded quantitative, 55 (0.69 g, 0.66 mmol) as an off-white solid. LC-MS M--H 1035.6 (CHNOS+H, calc: 1035.5). Compound Q was used directly in the next reaction without further purification. Preparation of Compound R A solution of Compound Q (0.69 g, 0.66 mmol) in 1,4- 60 dioxane (10 mL) was treated with hydrogen chloride (4.0 M solution in 1,4-dioxane, 10 mL). After 1 h, most of the 1,4- dioxane was removed under vacuum until ~2 mL remained. To this solution was added EtO (~100 mL). The product precipitated as an HCl salt. The precipitate was washed with 65 ether and dried under vacuum to afford crude Compound R. Compound KC-9 was prepared following the method yield exceeded quantitative, (0.67 g. 0.66 mmol) as an off described in Example 3 to prepare N-(oxycodone-6-enol US 8,685,916 B2 117 118 carbonyl)piperidine-2-methylamine (Compound KC-11), carbonyl)piperidine-2-methylamine-L-arginine-L-alanine but using pyrrolidine-2-yl-methylcarbamic acid tert-butyl acetate (Compound KC-13), but using mono-tert-butyl mal ester instead of piperidine-2-yl-methylcarbamic acid tert-bu onate instead of Boc-Ala-OH. LC-MS M+H 698.4 tyl ester. LC-MS M+H 442.1 (CHNO+H, calc: (C.H.N.O+H, calc: 698.7). 442.3). Example 5 Example 7

Synthesis of N-(oxycodone-6-enol-carbonyl)pyrroli Synthesis of N-(oxycodone-6-enol-carbonyl)piperi dine-2-methylamine-L-arginine-malonate (Com 10 pound KC-10) dine-2-methylamine-L-arginine-glycine-acetate (Compound KC-14)

KC-10 15

KC-14

OH

N N ls NH 25 N NH2

30 Compound KC-10 was prepared following the method described in Example 3 to prepare N-(oxycodone-6-enol Compound KC-14 was prepared following the method carbonyl)piperidine-2-methylamine-L-arginine-L-alanine described in Example 3 to prepare N-(oxycodone-6-enol acetate (Compound KC-13), but using pyrrolidine-2-yl-me carbonyl)piperidine-2-methylamine-L-arginine-L-alanine thylcarbamic acid tert-butyl ester instead of piperidine-2-yl 35 acetate (Compound KC-13), but using Boc-Gly-OH instead methylcarbamic acid tert-butyl ester, and using mono-tert of Boc-Ala-OH. LC-MS M+H 711.3 (CHNO+H, butylmalonate instead of Boc-Ala-OH. LC-MSM-I-H 684.4 calc: 711.4). (CHNO+H, calc: 684.4). Example 6 40 Example 8 Synthesis of N-(oxycodone-6-enol-carbonyl)piperi dine-2-methylamine-L-arginine-malonate (Com Synthesis of N-(oxycodone-6-enol-carbonyl)piperi pound KC-12) dine-2-methylamine-L-arginine-L-alanine-malonate 45 (Compound KC-15)

KC-12 KC-15 50

O OH Os N O n O H O O H N () is lusu lulls N 55 \ HQ N 1N OH H N O O % NH NH2

H-- 2 60

Compound KC-15 was prepared following the method 65 described in Example 3 to prepare N-(oxycodone-6-enol Compound KC-12 was prepared following the method carbonyl)piperidine-2-methylamine-L-arginine-L-alanine described in Example 3 to prepare N-(oxycodone-6-enol acetate (Compound KC-13), but using mono-tert-butyl mal US 8,685,916 B2 119 120 onate instead of acetic anhydride. LC-MS M+H 769.6 butyl malonate instead of acetic anhydride. LC-MSM-H (C2HNO+H, calc: 769.4). 755.5 (CHNO+H, calc: 755.4). Example 9 Example 11

Synthesis of N-(oxycodone-6-enol-carbonyl)piperi Synthesis of N-(oxycodone-6-enol-carbonyl)-R-(pip dine-2-methylamine-L-arginine-glycine-malonate eridine-2-methylamine) (Compound KC-18) (Compound KC-16) 10

KC-18 KC-16

15 lulO O NNH 1NH OH O

N NH H 2

25

Compound KC-16 was prepared following the method described in Example 3 to prepare N-(oxycodone-6-enol Compound KC-18 was prepared following the method carbonyl)piperidine-2-methylamine-L-arginine-L-alanine 30 described in Example 3 to prepare N-(oxycodone-6-enol acetate (Compound KC-13), but using Boc-Gly-OH instead carbonyl)piperidine-2-methylamine (Compound KC-11), of Boc-Ala-OH, and using mono-tert-butyl malonate instead but using (R)-piperidine-2-yl-methylcarbamic acid tert-butyl of acetic anhydride. LC-MSM-I-H 755.4 (CHNO+H, ester instead of (R.S)-piperidine-2-yl-methylcarbamic acid calc: 755.4). 35 tert-butyl ester. LC-MSM--H456.2 (CHNO+H, calc: 456.3). Example 10 Example 12 Synthesis of N-(oxycodone-6-enol-carbonyl)-R-(pip- 40 eridine-2-methylamine)-L-arginine-glycine-malonate Synthesis of N-(hydrocodone-6-enol-carbonyl)-R- (Compound KC-17) (piperidine-2-methylamine)-L-arginine-glycine-mal onate (Compound KC-31)

45 KC-17

KC-31

O O 50 NH lul O O 1NH OH NH lul O 1NH OH O .N. NH2 55 H N NH H 2

60 Compound KC-17 was prepared following the method described in Example 3 to prepare N-(oxycodone-6-enol Compound KC-31 was prepared following the method carbonyl)piperidine-2-methylamine-L-arginine-L-alanine described in Example 10 to prepare N-(oxycodone-6-enol acetate (Compound KC-13), but using (R)-piperidine-2-yl carbonyl)-R-(piperidine-2-methylamine)-L-arginine-gly methylcarbamic acid tert-butyl ester instead of (R.S)- 65 cine-malonate (Compound KC-17), except hydrocodone was piperidine-2-yl-methylcarbamic acid tert-butyl ester, using used instead of oxycodone. LC-MS M+H 739.6 Boc-Gly-OH instead of Boc-Ala-OH, and using mono-tert (CHNO+H calc: 739.9).

US 8,685,916 B2 123 124 -continued

r O H O N OH NH YY O O NH

s H

TP-1

Preparation of Compound U (Cas H.N.OS+H, calc: 870.5). Compound X was used A solution of 2-(tert-butoxycarbonylamino-methyl)-pip directly in the next reaction without further purification. eridine-1-carboxylic acid benzyl ester (Compound T) (3.0 g, Preparation of Compound Y 8.61 mmol) was treated with HCl (4.0 M solution in 1,4- A solution of Compound X (1.7 g, 1.94 mmol) in 1,4- dioxane, 20 mL) for 1 h. The solvents were removed under dioxane (10 mL) was treated with HCl (4.0 M solution in vacuum untila volume of ~10 mL remained, after which EtO 1,4-dioxane, 10 mL) for 30 min. The solvents were then (500 mL) was added. The precipitate was filtered off and removed until a volume of -5 mL was reached, after which washed with EtO (2x100 mL) and dried to afford crude 25 EtO was added (250 mL). The resulting precipitate was Compound U in a quantitative yield (2.87 g., 8.61 mmol) as a filtered off, washed with EtO (2x75 mL), and dried to afford white solid. LC-MS IM+H 249.3 (CHHO-H, calc: crude Compound Y. The crude compound was dissolved in 249.3). Compound U was used directly in the next reaction water (15 mL), and the solution was subjected to HPLC without further purification. purification. Nanosyn-Pack Microsorb (100-10) C-18 col Preparation of Compound V 30 umn (50x300mm); flow rate: 100 mL/min: injection volume A solution of Boc-Arg(Pbf)-OH (4.54 g, 8.61 mmol), 15 mL. mobile phase A: 100% water, 0.1% TFA; mobile Compound U (2.87g, 8.61 mmol), and DIEA (3.9 mL, 22.4 phase B: 100% ACN, 0.1%TFA: isocratic elution at 0% B in mmol) in DMF (100 mL) was cooled to 0°C. (in an ice bath): 5 min, gradient elution from 0% to 30% B in 30 min, isocratic HATU (3.5g, 8.61 mmol) was added in portions over 15 min. elution at 30% B in 20 min, gradient elution florin 30% B to The reaction mixture was raised to ambient temperature, and 35 50% B in 40 min; detection at 254 nm. Fractions containing stirring was continued for an additional 2 h. DMF was then the desired product were combined and concentrated under removed under vacuum, and the residue was diluted with vacuum. The residue was dissolved in MeCN (-2 mL) and 0.1 EtOAc (500 mL) and extracted with water (3x100 mL) and N HCl (-8 mL) and lyophilized overnight to provide Com brine (100 mL). The organic layer was dried over NaSO and pound Y in 53% yield (0.84g, 1.00 mmol) as a foamy solid. filtered. The removal of the solvents under vacuum yielded 40 LC-MSM-I-H 770.4 (CHN.OS+H, calc: 770.5). the crude Compound V which was purified by flash chroma Preparation of Compound Z tography using CHCl and MeOH to afford Compound V in To a solution of Compound Y (0.75 g, 0.89 mmol), mono 45% yield (2.9 g, 3.83 mmol) as a foamy solid. LC-MS tert-Butyl Malonate (0.13 mL, 1.08 mmol), and DIEA (0.46 M+H 757.6 (ClsHNOS+H, calc: 757.4). mL, 2.7 mmol) in DMF (15 mL) at 5° C. was added BOP Preparation of Compound W 45 (0.39 g, 1.08 mmol) in portions. The reaction mixture was A solution of Compound V (2.7 g, 3.57 mmol) in MeOH stirred at ambient temperature for 1 h. DMF was removed (100 mL) was treated with palladium (5 wt.% on activated under vacuum, and the residue was diluted with EtOAc (100 carbon, 0.6 g) and subjected to hydrogenation at 70 psi for 1 mL), and washed with water (2x50 mL) and brine (25 mL). h. The reaction mixture was filtered using a Celite pad. The The organic layer was dried over NaSO and filtered; the removal of MeOH afforded Compound Win 99% yield (2.19 50 removal of the solvents afforded Compound Z in quantitative g, 3.51 mmol) as a foamy solid. LC-MS M+H 623.6 yield (1.2g, 0.89 mmol) as an oil. LC-MS M+H 912.8 (CHNOS+H, calc: 623.4). Compound W was used (C.H.N.OS+H, calc: 912.5). Compound Z was used directly in the next reaction without further purification. directly in the next reaction without further purification. Preparation of Compound X To a solution of tapentadol hydrochloride (0.5 g, 1.94 55 Preparation of N-(Tapentadol-carbonyl)piperidine-2- mmol) and DIEA (0.34 mL, 1.94 mmol) in CHCl (15 mL) methylamine-L-arginine-malonate (Compound was added 4-nitrophenyl chloroformate (0.38g, 1.89 mmol), TP-5) and the reaction mixture was sonicated for 30 min. To this reaction mixture was added Compound W (1.18 g, 1.89 A solution of Compound Z (1.2g, 0.89 mmol) in TFA (10 mmol) in DMF (5 mL) at 5°C. The resultant reaction mixture 60 mL) was treated with 5% m-cresol for 1 h. The product was was warmed to ambient temperature, and then allowed to stir precipitated via addition of EtO (100 mL). The precipitate for 2 h. The solvents were then removed under vacuum, and was washed with EtO (2x100 mL) and dried under vacuum. the residue was diluted with EtOAc (100 mL), and washed The resultant product was dissolved in water (15 mL), and the with water (2x50 mL) and brine (25 mL). The organic layer solution was subjected to HPLC purification. Nanosyn-Pack was dried over NaSO and filtered; the removal of the sol 65 Microsorb (100-10) C-18 column (50x300 mm); flow rate: vents under vacuum afforded Compound X in quantitative 100 mL/min: injection volume 15 mL, mobile phase A: 100% yield (1.7 g, 1.94 mmol) as an oil. LC-MS M+H 870.8 water, 0.1% TFA: mobile phase B: 100% ACN, 0.1% TFA: US 8,685,916 B2 125 126 gradient elution from 0% to 20% B in 30 min, isocratic carbonyl)-R-(piperidine-2-methylamine)-L-arginine-gly elution at 20% B in 30 min, gradient elution from 20% B to cine-malonate (Compound KC-17), except using acetic 45% B in 35 min; detection at 254 nm. Fractions containing anhydride instead of mono-tert-butyl malonate and using the desired product were combined and concentrated under hydrocodone instead of oxycodone. LC-MS IM+H 695.8 vacuum. The residue was dissolved in MeCN (-2 mL) and 0.1 (CHNO+H calc: 695.8). N HCl (-8 mL) and lyophilized overnight to provide Com pound TP-5 in 88% yield (0.56g, 0.79 mmol, 95.0% purity) Example 16 as a foamy solid. LC-MS M+H 604.5 (CHN,O+H, calc: 604.4). Synthesis of N-(hydrocodone-6-enol-carbonyl)-R- 10 (piperidine-2-methylamine)-L-arginine-L-alanine Example 14 acetate (Compound KC-37) Synthesis of N-(hydrocodone-6-enol-carbonyl)-R- (piperidine-2-methylamine)-L-arginine-L-alanine malonate (Compound KC-35) 15 KC-37

KC-35

O O N. H lul 25 1NH OH O NH s H 2 30 Compound KC-37 was prepared following the method described in Example 10 to prepare N-(oxycodone-6-enol Compound KC-35 was prepared following the method carbonyl)-R-(piperidine-2-methylamine)-L-arginine-gly described in Example 10 to prepare N-(oxycodone-6-enol 35 cine-malonate (Compound KC-17), except using acetic carbonyl)-R-(piperidine-2-methylamine)-L-arginine-gly anhydride instead of mono-tert-butyl malonate, using Boc cine-malonate (Compound KC-17), except using Boc-Ala Ala-OH instead of Boc-Gly-OH and using hydrocodone OH instead of Boc-Gly-OH and using hydrocodone instead instead of oxycodone. LC-MS M+H 695.8 of oxycodone. LC-MS M+H 753.7 (CHNO+H calc: (CHNO,+H calc: 695.8). 753.9). 40 Example 17 Example 15 Synthesis of N-(hydrocodone-6-enol-carbonyl)-R- Synthesis of N-(hydrocodone-6-enol-carbonyl)-R- (piperidine-2-methylamine)-L-arginine-acetate (piperidine-2-methylamine)-L-arginine-glycine-ac (Compound KC-38) etate (Compound KC-36) 45

KC-38 KC-36 50

55

60

65 Compound KC-36 was prepared following the method Compound KC-38 was prepared following the method described in Example 10 to prepare N-(oxycodone-6-enol described in Example 10 to prepare N-(oxycodone-6-enol US 8,685,916 B2 127 128 carbonyl)-R-(piperidine-2-methylamine)-L-arginine-gly- cine-malonate (Compound KC-17), except employing ethyl cine-malonate (Compound KC-17), except not employing 3-((tert-butoxycarbonylamino)methyl)piperazine-1-car Boc-Gly-OH, using acetic anhydride instead of mono-tert- boxylate (compound CC, see Example 20 for synthesis) butyl malonate, and using hydrocodone instead of oxyc- instead of piperidine-2-yl-methylcarbamic acid tert-butyl odone. LC-MSM-H 638.5 (CHNO+H calc: 638.7). 5 ester, using acetic anhydride instead of mono-tert-butyl mal onate, and using hydrocodone instead of oxycodone. LC-MS Example 18 M+H 768.7 (CHNO+H calc: 768.9). Synthesis of N-(hydrocodone-6-enol-carbonyl)-R- Example 20 (piperidine-2-methylamine)-L-arginine-malonate 10 (Compound KC-39) Synthesis of Ethyl 3-((tert-butoxycarbonylamino) methyl)piperazine-1-carboxylate (Compound CC)

KC-39 15

O OH N 1. N 1 or D -esBoc.O 9 r D ls. AA H2/Pd, Pt/C, 11.2 trichloroethane so N- N Compound KC-39 was prepared following the method 1. Hel J. H carbonyl)-R-(piperidine-2-methylamine)-L-arginine-gly-described in Example 10 to prepare N-(oxycodone-6-enol- OY irN N D (EtOCO)2O OY irN N D oHCI cine-malonate (Compound KC-17), except not employing 35 N N Boc-Gly-OH and using hydrocodone instead of oxycodone. -N BB H LC-MSM-H 682.7 (CHNO+H calc: 682.8). 11so CC Example 19 40 Synthesis of N-(hydrocodone-6-enol-carbonyl)-N- Synthesis of pyrazin-2-ylmethyl-carbamic acid ethyl-(2-methylamino)piperazine-4-carboxylate-L- tert-butyl ester (Compound AA) arginine-glycine-acetate (Compound KC-42) To a solution of 2-aminomethyl-pyrazine (5.0 g, 45.87 45 mmol) in isopropanol (50 ml) was added di-tert-butyl-pyro carbonate (12.0 g, 55.84 mmol); the mixture was stirred at KC-42 ambient temperature for 2h. Next, the solvent was evaporated and the residue was dissolved in DCM (30 ml), and subjected to silica gel purification (chloroform/methanol gradient 50 0->30% in 100 min.). Fractions containing desired product were combined and evaporated. Residue was dried under vacuum to provide Compound AA in quantitative yield (10.22 g, 99% purity) as amorphous solid. LC-MS: (m/z) observed (M+H") 210.5 (CHNO+H calc: 210.3). 55 Synthesis of piperazin-2-ylmethyl-carbamic acid tert-butyl ester (Compound BB) Compound AA (10.22 g, 45.87 mmol) was dissolved in 60 methanol (350 ml) followed by the addition of 1,1,2-trichlo roethane (9.39 ml, 100.91 mmol), a suspension of palladium on activated carbon (10 wt.%, 488 mg) and platinum on activated carbon (10 wt.%. 897 mg) in water (20 ml). The mixture was Subjected to hydrogen (70 psi) on a Parr appa Compound KC-42 was prepared following the method 65 ratus at ambient temperature for 3h. The reaction mixture was described in Example 10 to prepare N-(oxycodone-6-enol- then filtered through the pad of celite and the filtrate was carbonyl)-R-(piperidine-2-methylamine)-L-arginine-gly- separated and evaporated. The residue was re-dissolved in US 8,685,916 B2 129 130 isopropanol and re-evaporated. The resulting solid was dried Example 22 under vacuum at ambient temperature overnight to provide the hydrochloric salt of Compound BB in quantitative yield Synthesis of N-(hydrocodone-6-enol-carbonyl)-N- (14.05 g, 99% purity) as off-white solid. LC-MS. (m/z) ethyl-(2-methylamino)piperazine-4-carboxylate-L- observed (M+H") 216.5 (CHHO-H calc: 216.3). The arginine-malonate (Compound KC-44) desired product was used directly without further purifica tion.

KC-44 Synthesis of Ethyl 3-((tert-butoxycarbonylamino) 10 methyl)piperazine-1-carboxylate (Compound CC)

To a solution of Compound BB (7.00g, 22.79 mmol) in EtOH (abs., 200 ml) was added diethyl pyrocarbonate (3.35 ml, 22.79 mmol) portionwise (330 ulx10) over 5 min: The 15 reaction was monitored via LC-MS until complete consump tion of the starting material. Upon completion, the formed precipitate was filtered and discarded. The filtrate was evapo rated until dryness and was then dried under vacuum to pro vide hydrochloric salt of Compound CC in 94% yield (6.75 g, 99% purity) as an off-white solid. LC-MS: (m/z) observed (M+H") 288.2 (CHNO+H calc: 288.4). 25 Example 21 Compound KC-44 was prepared following the method Synthesis of N-(hydrocodone-6-enol-carbonyl)-N- described in Example 10 to prepare N-(oxycodone-6-enol ethyl-(2-methylamino)piperazine-4-carboxylate-L- 30 carbonyl)-R-(piperidine-2-methylamine)-L-arginine-gly arginine-acetate (Compound KC-43) cine-malonate (Compound KC-17), except using ethyl 3-((tert-butoxycarbonylamino)methyl)piperazine-1-car boxylate (compound CC, see Example 20 for synthesis) instead of piperidine-2-yl-methylcarbamic acid tert-butyl KC-43 35 ester, not employing Boc-Gly-OH, and using hydrocodone

instead of oxycodone. LC-MS M+H 755.5 (CHNO+H calc: 755.8). Example 23 40 Synthesis of N-(hydrocodone-6-enol-carbonyl)-N- ethyl-(2-methylamino)piperazine-4-carboxylate-L- arginine-glycine-malonate (Compound KC-45)

45

KC-45

50

55 Compound KC-43 was prepared following the method described in Example 10 to prepare N-(oxycodone-6-enol carbonyl)-R-(piperidine-2-methylamine)-L-arginine-gly cine-malonate (Compound KC-17), except using ethyl 3-((tert-butoxycarbonylamino)methyl)piperazine-1-car 60 boxylate (Compound CC, see Example 20 for synthesis) instead of piperidine-2-yl-methylcarbamic acid tert-butyl Compound KC-45 was prepared following the method ester, not employing Boc-Gly-OH, using acetic anhydride described in Example 10 to prepare N-(oxycodone-6-enol instead of mono-tert-butyl malonate, and using hydrocodone 65 carbonyl)-R-(piperidine-2-methylamine)-L-arginine-gly instead of oxycodone. LC-MS M+H 711.7 cine-malonate (Compound KC-17), except using ethyl (C5H5NO+H calc: 711.8). 3-((tert-butoxycarbonylamino)methyl)piperazine-1-car US 8,685,916 B2 131 132 boxylate (compound CC, see Example 20 for synthesis) instead of piperidine-2-yl-methylcarbamic acid tert-butyl ester and using hydrocodone instead of oxycodone. LC-MS M+H 812.8 (CHNO+H calc: 812.9). Example 24 Synthesis of N-(hydrocodone-6-enol-carbonyl)-(2- methylamino)piperidine-4-carboxylate-L-arginine glycine-acetate (Compound KC-40) 10

US 8,685,916 B2 139 140 Preparation of Compound DD Preparation of Compound HH 2-Bromo isonicotinic acid (20.2 g, 100 mmol) was dis A solution of hydrocodone-free base (1.90 g. 6.35 mmol) solved in DMF (500 mL) at ambient temperature. CsCO in THF (50 mL) was cooled to -78° C. and then 0.5M toluene (32.6 g. 100 mmol) was added in one portion, followed by solution of KHMDS (12.7 mL, 6.35 mmol) was added drop MeI (6.3 mL, 100 mmol). The mixture was stirred at ambient wise over 5 min under nitrogen. The reaction mixture was temperature for 15 h, followed by the addition of water (500 stirred for 30 min, and then added to a solution of 4-nitrophe mL). The mixture was extracted with EtOAc (500 mL). The nyl chloroformate (1.35 g. 6.35 mmol) in THF (25 mL) drop organic layer was washed with water (500 mL), brine (500 wise over 5 min under nitrogen and cooling with dry icef mL) and then dried over NaSO. The organic layer was then acetone. Upon completion, 2 M HCl in diethyl ether (25 mL) filtrated and concentrated to give compound DD as a white 10 solid in 80% yield (17.4g, 80.5 mmol). LC-MS. M+H 217.0 and ether (100 mL) was added dropwise to the reaction mix (C.HBrNO+H, calc: 216.1). Compound DD was used ture to produce a fine white precipitate. The precipitate was directly without further purification. filtered on a glass frit and washed with ether (3x50 mL). The Solid was dried under high vacuum overnight, then dissolved Preparation of Compound EE 15 Compound DD (17.4g, 80.5 mmol) was dissolved in DMF in 5%aq KHPO solution (200 mL) and extracted with DCM (160 mL), followed by the addition of Zn(CN), (5.7g, 48.53 (2x50 mL). The organic phase was dried over NaSO (anh.), mmol) in one portion. The mixture was degassed using nitro filtered, and the solvent was concentrated under vacuum to gen and then Pd(PPh3)4 (4.7 g) was added. The mixture was the volume -10 mL. To the mixture was added 2 M solution degassed again and then heated in an oil bath (120° C.). After of HCl in diethyl ether (20 mL) and ether (100 mL). The 2.5 h, the reaction was cooled to ambient temperature and resulting fine white precipitate was filtered off, washed with water (200 mL) was added. The mixture was stirred for 30 ether (2x50 mL) and dried under high vacuum to afford min and then filtered through a frit. The solid collected was compound HH in 66.1% yield (2.1 g, 4.20 mmol). LC-MS washed with water (2x100 mL) and then dried under vacuum M+H: 465.3 (CHNO+H, calc: 464.2). Retention time to give compound EE in 84% yield (11.0 g. 67.9 mmol). 25 Chromolith Speed Rod RP-18e C18 column (4.6x50 mm); LC-MS. M+H 163.2 (CHNO+H, calc: 162.1). Com flow rate: 1.5 mL/min: mobile phase A: 0.1% TFA/water; pound EE was used directly without further purification. mobile phase B 0.1%TFA/ACN:gradient elution from 5% B Preparation of Compound FF to 100% B over 9.6 min, detection 254 nm: 4.94 min. Compound EE (20.0g, 123.4 mmol) was dissolved in IPA 30 Preparation of Compound II (500 mL). BocO (37.7 g, 172.8 mmol), Pd/5% on barium Compound GG (6.0 g, 22.0 mmol) and compound HH sulfate (6.0 g) and NEt (35 mL. 246.9 mmol) were added to (12.5g, 24.0 mmol) were dissolved in DMF (40 mL) and the reaction mixture. The mixture was hydrogenated at 55 psi DIEA (15.3 mL. 88 mmol) was added. The reaction mixture for 4h on a Parr hydrogenator. The mixture was then filtered was stirred at 40° C. for approximately 4 h, until all the through a celite pad and then the celite pad was washed with 35 MeOH (3x80 mL). The combined filtrate was then concen starting amine GG was consumed. Upon reaction completion, trated and the residue was partitioned between EtOAc (300 the DMF was evaporated and the resulting oily product was mL) and water (100 mL). The organic layer was washed with dissolved in DCM (700 mL). The mixture was then washed 10% citric acid (50 mL) and brine (50 mL), followed by with 5% sodium phosphate (2x700 mL), 0.1 Naq. HCl (500 drying over NaSO. Next the mixture was filtered and con 40 mL) and brine (750 mL). The organic phase was dried over centrated to afford compound FF in 86% yield (28.0 g, 106.8 NaSO (anhydrous), filtered and the solvent was evaporated. mmol). LC-MS. M+H 267.4 (CHNO+H, calc: 266.5). The oily product was dried in high vacuum overnight to afford Compound FF was used directly without further purification. compound II in 91.2% yield. (12.2g, 20.1) LC-MS: M-H Preparation of Compound GG 578.6 (CHNO+H, calc: 578.7). Retention time Chro To a solution of compound FF (1.50 g, 5.64 mmol) in 45 molith Speed Rod RP-18e C18 column (4.6x50 mm); flow MeOH (25 mL) was carefully added, under nitrogen, 10% rate: 1.5 mL/min: mobile phase A: 0.1% TFA/water, mobile Pd/C (250mg), 10% PBC (200mg) and 1,1,2-trichloroethane phase B 0.1% TFA/ACN; gradient elution from 5% B to (630 mL, 6.8 mmol. 1.2 eq). The reaction mixture was stirred 100% B over 9.6 min, detection 254 nm): as 4 isomers, 5.90 at 65 psi overnight. Upon completion, the reaction mixture min(A), 5.94 min(A), 6.47 min (B), 6.58 min (C). was filtered through a Celite-padded glass frit and washed 50 Preparation of Compound JJ (Major Isomers) with MeOH (3x20 mL). The filtrate was concentrated under A solution of compound II (12.2g, 21.2 mmol) in DCM vacuum to the volume ~10 mL and diethyl ether (100 mL) (100 mL) was treated with 4M solution of hydrogen chloride was added. The resulting fine white precipitate was filtered, in 1,4-dioxane (50 mL). After 1 h, solvent was removed under washed with ether (2x50 mL) and dried under high vacuum. 55 vacuum until about ~50 mL remained. Diethyl ether (-500 The resulting HCl salt was dissolved withsonication in water mL) was added to the reaction mixture, which produced a fine (30 mL) and aqueous 1 NNaOH solution (10 mL) was added. white precipitate. The precipitate was filtered off, washed The reaction mixture was extracted with DCM (3x25 mL). with ether (3x150 mL) and dried under vacuum to give the The organic phase was dried over anhydrous Na2SO4, fil HCl salt of compound JJ as a fine white solid. The solid was tered, the solvent was evaporated under vacuum and the oily 60 product was dried under high vacuum overnight. This dissolved in water (70 mL) and acetic acid (10 mL) and the afforded Compound GG (1.08 g., 72.5%). LC-MS. M+H solution was subjected to HPLC purification, 5 runs: Nano 267.2 (CH NO+H, calc: 267.1). Retention time Chro syn-Pack Microsorb (100-10) C-18 column (50x300 mm); molith Speed Rod RP-18e C18 column (4.6x50 mm); flow flow rate: 100 mL/min: injection volume: 4x5 mL. mobile rate: 1.5 mL/min: mobile phase A: 0.1% TFA/water, mobile 65 phase A: 100% water, 0.1% TFA; mobile phase B: 100% phase B 0.1% TFA/ACN; gradient elution from 5% B to acetonitrile, 0.1%TFA:gradient elution from 5% to 30% B in 100% B over 9.6 min): 2.79 min. 60 min; detection at UV 254 nm. Fractions containing the US 8,685,916 B2 141 142 major isomers were combined and concentrated under night to afford compound MM in 93.8% yield (22.9 g, 23.9 vacuum. The resulting oily residue was co-evaporated with mmol) as a yellow oil. LC-MS: M+H 1005.7 isopropanol (3x100 mL). The oily product was treated with 2 (CHNOS+H, calc: 1005.2). Retention time Chro M HCl in ether (20 mL) and ether (400 mL) to produce a fine molith Speed Rod RP-18e C18 column (4.6x50 mm); flow white precipitate. The precipitate was filtered off, washed rate: 1.5 mL/min: mobile phase A: 0.1% TFA/water, mobile with ether (2x50 mL) and dried under high vacuum to afford phase B 0.1% TFA/ACN; gradient elution from 5% B to the two major isomer of compound JJ in 69.2% yield (8.1 g, 100% B over 9.6 min, detection 254 nm): 4.75 min. 14.7 mmol). LC-MS, M+H 498.4 (CHNO+H, calc: Preparation of Compound NN (Major Isomers) 498.6). Retention time Chromolith Speed Rod RP-18e C18 10 To a solution of Compound MM (22.8 g. 22.9 mmol) in column (4.6x50mm); flow rate: 1.5 mL/min: mobile phase A: methanol (120 mL) at 5° C. was added aqueous solution of 0.1% TFA/water; mobile phase B 0.1% TFA/ACN; gradient LiOH (1.6 g. 70 mmol) in water (50 mL). The temperature of elution from 5% B to 100% B over 9.6 min, detection 254 nm: as 2 isomers: 2.81 min (major-1), 2.96 min (major-2). the reaction mixture was raised to ambient temperature and Preparation of Compound KK (Major Isomers) 15 stirring was continued for an additional 2h. Upon the reaction To a solution of Boc-Arg(Pbf)-OH (18.96 g., 36.0 mmol), completion, the reaction mixture was neutralized with acetic Compound JJ (19.6 g., 34.3 mmol) and HATU (13.3 g, 37.7 acid to pH -4.0 and the methanol was evaporated under mmol) in DMF (200 mL) at 5°C. was added DIEA (24.0 mL, vacuum. The resulted solution was subjected to prep HPLC 137 mmol) dropwise over 5 min. The temperature of the purification. Nanosyn-Pack Microsorb (100-10) C-18 col reaction mixture was raised to ambient temperature and stir umn (50x300mm); flow rate: 100 mL/min: injection volume: ring was continued for an additional hour. Upon reaction 4x5 mL. mobile phase A: 100% water, 0.1% TFA; mobile completion, DMF was removed under vacuum and the reac phase B: 100% acetonitrile, 0.1%TFA; gradient elution from tion mixture was then diluted with DCM (300 mL), washed 5% to 30% B in 20 min; isocratic 30% B in 15 min, 30% to with 2% aq. HSO (500 mL), then with 5% sodium phos 25 65% in 25 min; detection at UV 254 nm. Fractions containing phate (500 mL) and brine (750 mL). The organic phase was the pure product were combined and concentrated under dried over NaSO (anh.), filtered, and the solvent was evapo vacuum. The resulting oily residue was co-evaporated with rated under vacuum. The oily product was dried under high toluene (3x100 mL). The oily product was dried under high vacuum to give compound NN in 54.9% yield (12.4g, 12.6 vacuum overnight to afford compound KK as a foamy solid in 30 97.2% yield. (34.2 g, 33.3 mmol) LC-MS. M+H 1007.1 mmol). LC-MS, M+H 991.7 (CHNOS+H, calc: (CH, N,OS+H, calc: 1007.2). Retention time Chro 991.5). Retention time Chromolith Speed Rod RP-18e C18 molith Speed Rod RP-18e C18 column (4.6x50 mm); flow column (4.6x50mm); flow rate: 1.5 mL/min: mobile phase A: rate: 1.5 mL/min: mobile phase A: 0.1% TFA/water, mobile 0.1% TFA/water; mobile phase B 0.1%TFA/ACN; gradient phase B 0.1% TFA/ACN; gradient elution from 5% B to 35 elution from 5% B to 100% B over 9.6 min, detection 254 100% B over 9.6 min, detection 254 nm): 5.43 min. nm: 4.59 min. Preparation of Compound LL (Major Isomers) Preparation of N-(hydrocodone-6-enol-carbonyl)- To a solution of compound KK (34.3 g, 34.0 mmol) in (2-methylamino)piperidine-4-carboxylate-L-argin DCM (100 mL) was added 4.0 M solution of hydrogen chlo 40 ine-glycine-acetate (Compound KC-40, Major Iso ride in 1,4-dioxane (150 mL). After 1 h, the solvent was mers) evaporated under vacuum to about 50 mL and to the reaction mixture was added diethyl ether (500 mL) to produce a fine Compound NN (7.9 g, 7.9 mmol) was dissolved in TFA (50 white precipitate. The precipitate was filtered off, washed mL) and was stirred for 1 h. Next the TFA was evaporated with ether (3x150 mL) and dried under vacuum to give the 45 under vacuum and the resulting oily residue was dissolved in HCl salt of compound LL in 82.7% yield (23.9 g, 28.1 mmol) acetic acid/DCM (10 mL/10 mL) and treated with 2 M HCl/ as a fine white solid. LC-MS, M+H 906.6 (CHN,OS+ ether. The formed white precipitate was filtered off and H, calc: 906.1). Retention time Chromolith SpeedRod washed with ether (2x50 mL). The solid was dissolved in RP-18e C18 column (4.6x50 mm); flow rate: 1.5 mL/min: water (60 mL) and subjected to prep HPLC purification. mobile phase A: 0.1% TFA/water; mobile phase B 0.1% 50 Nanosyn-Pack Microsorb (100-10) C-18 column (50x300 TFA/ACN; gradient elution from 5% B to 100% B over 9.6 mm); flow rate: 100 mL/min: injection volume: 4x5 mL.; min, detection 254 nm: 4.46 min. mobile phase A: 100% water, 0.1% TFA; mobile phase B: Preparation of Compound MM (Major Isomers) 100% acetonitrile, 0.1% TFA; gradient elution from 5% to To a solution of HO-Gly-NAc (3.0g, 25.5 mmol), Com 55 30% B in 60 min; detection at UV 210 nm. Fractions contain pound LL (23.4g, 24.3 mmol. 1 eq) and HATU (9.7 g. 25.5 ing the desired product were combined and concentrated mmol) in DMF (100 mL) at 5° C. was added DIEA (17 mL, under vacuum. The resulting oily residue was co-evaporated 100 mmol) dropwise over 5 min. The temperature of the with toluene (3x100 mL). The oily product was dried under reaction mixture was raised to ambient temperature and stir high vacuum to give a solid. The resulting Solid was dissolved ring was continued for an additional hour. Upon reaction 60 in 0.1 M HCl (50 mL) and lyophilized to give Compound completion, DMF was removed in high vacuum, and the KC-40 in 61.9% yield (3.2g, 4.9 mmol). LC-MS, M+H reaction mixture was diluted with DCM (300 mL), washed 739.7 (CHNO+H, calc: 739.4). Retention time Chro with 2% aq. HSO (500 mL), then with 5% sodium phos molith Speed Rod RP-18e C18 column (4.6x50 mm); flow phate (500 mL) and brine (750 mL). The organic phase was 65 rate: 1.5 mL/min: mobile phase A: 0.1% TFA/water, mobile dried over NaSO (anh.), filtered and the solvent was evapo phase B 0.1% TFA/ACN; gradient elution from 5% B to rated. The oily product was dried under high vacuum over 100% B over 9.6 detection 210 nm): 3.11 min. US 8,685,916 B2 143 144 Example 25 Example 27 N-(hydrocodone-6-enol-carbonyl)-(2-methylamino) N-(hydrocodone-6-enol-carbonyl)-(2-methy piperidine-3-carboxylate-L-arginine-glycine-acetate lamino)-N,N-dimethyl piperidine-4-carboxamide-L- (Compound KC-32) arginine-glycine-acetate (Compound KC-46)

KC-32 KC-46 10

15

25 Compound KC-46 was prepared following the method Compound KC-32 was prepared following the method described in Example 24 to prepare N-(hydrocodone-6-enol described in Example 24 to prepare N-(hydrocodone-6-enol carbonyl)-(2-methylamino)piperidine-3-carboxylate-L- carbonyl)-(2-methylamino)piperidine-3-carboxylate-L- 30 arginine-glycine-acetate (Compound KC-40), except con arginine-glycine-acetate (Compound KC-40), except using ducting a amide bond coupling of KC-40 with dimethyl methyl 6-bromonicotinate instead of methyl 2-bromoisomi amine using standard HATU coupling procedures (see Syn cothinate. LC-MS M+H 739.9 (CHNO+H calc: thesis of Compound O for a representative example). LC-MS 739.8). 35 M+H 766.6 (CHNO+H calc: 766.9). Example 26 Example 28 N-(hydrocodone-6-enol-carbonyl)-(2-methy lamino)-methyl piperidine-4-carboxylate-L-argin N-(hydrocodone-6-enol-carbonyl)-(2-methy ine-glycine-acetate (Compound KC-41) 40 lamino)-piperidine-4-carboxylate-L-arginine-gly cine-malonate (Compound KC-47)

KC-41 KC-47 O 45 ls O OH

O O 50 O O O NH lul N '^ ls 1NH OH O NH 55 st N NH H 2 H 2

60 Compound KC-47 was prepared following the method Compound KC-41 was prepared following the method described in Example 24 to prepare N-(hydrocodone-6-enol described in Example 24 to prepare N-(hydrocodone-6-enol carbonyl)-(2-methylamino)piperidine-3-carboxylate-L- carbonyl)-(2-methylamino)piperidine-3-carboxylate-L- arginine-glycine-acetate (Compound KC-40), except arginine-glycine-acetate (Compound KC-40), except not 65 employing Boc-Gly-OH instead of NAc-Gly-OH, followed employing LiOH (for methyl ester hydrolysis). LC-MS by Boc removal and coupling with mono-tert-butyl malonate M+H 753.7 (C.H.N.O+H calc: 753.9). (See synthetic procedures for the synthesis of compound