(12) United States Patent (10) Patent No.: US 8,343,927 B2 Mickle Et Al

USOO8343927B2

(12) United States Patent (10) Patent No.: US 8,343,927 B2 Mickle et al. (45) Date of Patent: *Jan. 1, 2013

(54) PHARMACEUTICAL COMPOSITIONS FOR (51) Int. Cl. PREVENTION OF OVERDOSE ORABUSE A638/00 (2006.01) A615 L/00 (2006.01) (75) Inventors: Travis Mickle, Charlottesville, VA (US); A638/04 (2006.01) Suma Krishnan, Blacksburg, VA (US); A638/06 (2006.01) A 6LX3L/785 (2006.01) James Scott Moncrief, Christiansburg, A6IP3/04 (2006.01) VA (US); Christopher Lauderback, A6IP 25/00 (2006.01) Blacksburg, VA (US) (52) U.S. Cl...... 514/18.1: 514/17.7: 514/1.3; 514/1.1; 530/330; 530/331; 424/1.69 (73) Assignee: Shire LLC, Florence, KY (US) (58) Field of Classification Search ...... None See application file for complete search history. (*) Notice: Subject to any disclaimer, the term of this patent is extended or adjusted under 35 (56) References Cited U.S.C. 154(b) by 0 days. This patent is Subject to a terminal dis U.S. PATENT DOCUMENTS claimer. 3,331,814 A 7, 1967 Randall et al. 3,676,492 A 7, 1972 Biel et al. 3,843,696 A 10/1974 Wagner et al. (21) Appl. No.: 12/881,008 3,846,399 A 11/1974 Hirschmann et al. 3,875,137 A 4, 1975 Jones et al. (22) Filed: Sep. 13, 2010 3,878,187 A 4, 1975 Schneider et al. 3,884,898 A 5, 1975 Schneider Prior Publication Data 3,975,342 A 8, 1976 Gross (65) 3,998,799 A 12/1976 Bodor et al. US 2011/0046226A1 Feb. 24, 2011 4,025,501 A 5, 1977 Leute 4,040,907 A 8, 1977 Ullman et al. 4,043,989 A 8, 1977 Schneider et al. Related U.S. Application Data 4,064.235 A 12/1977 Yanaihara et al. 4,064.236 A 12/1977 Dornet al. (63) Continuation of application No. 11/089,056, filed on 4,221,778 A 9/1980 Raghunathan Mar. 25, 2005, which is a continuation-in-part of 4,224.316 A 9/1980 Momany application No. 09/933,708, filed on Aug. 22, 2001, 4,242.256 A 12/1980 Sharpe et al. now abandoned, and a continuation-in-part of 4,297,346 A 10/1981 Rips et al. application No. 10/156,527, filed on May 29, 2002, (Continued) now Pat. No. 7,060,708, and a continuation-in-part of application No. 10/953,119, filed on Sep. 30, 2004, FOREIGN PATENT DOCUMENTS now Pat. No. 7.375,083, and a continuation-in-part of AU 541681.65 1, 1965 application No. 10/953,110, filed on Sep. 30, 2004, (Continued) now Pat. No. 7.338,939, and a continuation-in-part of application No. 10/923,257, filed on Aug. 23, 2004, OTHER PUBLICATIONS now Pat. No. 7,622,441, and a continuation-in-part of U.S. Appl. No. 12/698.323, entitled Amphetamine Prodrugs and application No. 10/923,088, filed on Aug. 23, 2004, Uses Thereof, filed Feb. 2, 2010. now Pat. No. 7,427,600, which is a Aggarwal, et al. Synthesis and Biological Evlaluation of Produgs of continuation-in-part of application No. Zidovudine, J. Med. Chem., 33(5):1505-1511 (1990). PCT/US03/05525, filed on Feb. 24, 2003, said Amidon G. et al. 5-Amino Acid Esters of Antiviral Nuclosides, Acyclovir, and AZT Are Absorbed by the Intestinal PEPT1 Peptide application No. 11/089,056 is a continuation-in-part of Transporter, Pharm Res, 16(2): 175 (1999), Abstract. application No. 10/955,006, filed on Sep. 30, 2004, Amidon, G. et al., A Theoretical Basis for a Biophamaceutic Drug now Pat. No. 7,169,752, and a continuation-in-part of Classification: The Corrleation of InVitro Drug Product Dissolution application No. 10/953,116, filed on Sep. 30, 2004, and InVivo Bioavailability, Pharmaceutical Research, vol. 12, No. 3 now Pat. No. 7.375,082, and a continuation-in-part of (1995). application No. 10/953,111, filed on Sep. 30, 2004, now abandoned, and a continuation-in-part of (Continued) application No. PCT/US2004/032131, filed on Sep. Primary Examiner — Maury Audet 30, 2004. (74) Attorney, Agent, or Firm — Blank Rome LLP (60) Provisional application No. 60/358,368, filed on Feb. 22, 2002, provisional application No. 60/366,258, (57) ABSTRACT filed on Mar. 22, 2002, provisional application No. The invention relates to pharmaceutical compositions com 60/358,381, filed on Feb. 22, 2002, provisional prised of a chemical moiety attached to an active agent in a application No. 60/567,800, filed on May 5, 2004, manner that substantially decreases the potential of the active provisional application No. 60/507.012, filed on Sep. agent to cause overdose or to be abused. When delivered at the 30, 2003, provisional application No. 60/567,802, proper dosage the pharmaceutical composition provides filed on May 5, 2004, provisional application No. therapeutic activity similar to that of the parent active agent. 60/568,011, filed on May 5, 2004, provisional application No. 60/362,082, filed on Mar. 7, 2002. 11 Claims, 86 Drawing Sheets US 8,343,927 B2 Page 2

U.S. PATENT DOCUMENTS 5,935,988 A 8, 1999 Matzke et al. 4,346,166 A 8/1982 Montag etal 5,935,995 A 8/1999 Bosslet et al. 4,358,604. A 1 1/1982 Albarella et al. 38.59 A 2:3: alyst al. 4,399,121 A 8/1983 Albarella et al. 5.955,105 A 9/1999 Mitraetal. 4,426,453 A 1/1984 Cree et al. 5,965,519 A 10/1999 Yatvin et al. 3.289 A ! E: Shiman et al. 5,965,695 A 10/1999 Simon et al. 4.483,807 A 1 1/1984 Asano et al. 3.8 A 2. f tal 4,489,080 A 12/1984 Lomen A 36 E." 4.489,165 A 12/1984 Wagner et al. 603094. A 2.2000 Summerton et al 4490,221. A 12/1984 Collange et al. 6,043,230 A 32000 Arimietal 4,552,864 A 1 1/1985 Antoni et al. 6048,736 A 42000 Kosak 4,569,844. A 2/1986 Jones 6,051.685. A 42000 Sakurada et al 4.584,398 A 4, 1986 Kuroiwa et al. F2000 K. tal 4,587,046 A 5/1986 Goodman et al. 8:3W - A 39: S.eronis et al. 3. A 33, et al. 6,093,391 A 7/2000 Kabanov et al. 4657.873 A 4, 1987 Avetal 6,146.658. A 1 1/2000 Bosslet et al. 4.730.04s A 3, 1988 Portoghese 6,235,718 B1 5/2001 Balasubramanium et al. 4,753,804. A 6/1988 Iaccheri et al. 855. R 239; S. et al. 4,776,121. A 10, 1988 Vicino 6,262,1074 - w B1 7/2001 Lietal.ashoua 4,801,575 A 1/1989 Pardridge 6,267.968 B1 7/2001 Bahr et al. 4,829,070 A 5/1989 Bodor 6,306,993 B1 10/2001 Rothbard et al. 33.6 A 99. Sri al 6,309.633 B1 10/2001 Ekwuribe et al. 4,925,673 A 5, 1990 Steiner et al. ge: R $383 ade 4,960,790 A 10/1990 Stella et al. 6.40737 B2 62002 Shashoua 4,976,962 A 12/1990 Bichon et al. 6.439323 B1 & 2002 Laietal. 3. A 88: Miyazaki et al. 6,458,842 B1 10/2002 Dickinson et al. 5,073,641 A 12, 1991 Bundgaard et al. 3. R. 39. Ret al. 5,087.616 A 2/1992 Myers etal Es sys- 5: 5.NEget al. 5, 120,859 A 6/1992 Webb 6,713,452 B2 3/2004 Ekwuribe et al. SE A 13: R al. 6,716.452 B1 4/2004 Piccariello et al. 5,196,107 A 3, 1993 Nakaoka et al. R 858: S. al. 1 5,219,5645,196.406 A 6/19933, 1993 ZalipskyKamei et etal. al. 6,818,659w B2 11/2004 Rajopadhyeackson et al. 4 K-1 6,846,831 B2 1/2005 Clemens 5,225,204 A 7/1993 Chen et al. 6,913,768 B2 7/2005 Couch et al. 5,233,025 A 8/1993 Miyazaki et al. 7,060,708 B2 * 6/2006 Piccariello et al...... 514,282 5,238,714 A 8/1993 Wallace et al. 7,089,140 B1 8/2006 McKenzie et al. 5,298.491. A 3/1994 Chauveau et al. 7,163,918 B2 1/2007 Piccariello et al. $335i A 88: E. et al 7,169,752 B2 * 1/2007 Mickle et al...... 514f13 5.378,712 A 1, 1995 Millet 7,338.939 B2 3/2008 Mickle et al...... 514f13 - w get al. 7,375,082 B2* 5/2008 Mickle et al. ... 514/13 3:1; A 892; list st al. 7,375,083 B2 * 5/2008 Mickle et al. ... 514/13 is A 332 Missal. 7,427,600 B2* 9/2008 Mickle et al...... 514/1.1 W - 7,438,900 B2 10/2008 Piccariello et al.

53. A 2. Best al 7.622,441 B2 * 1 1/2009 Mickle et al...... 514/1.1 5.539.95 A 6, 1996 RE 7,772,222 B2 8/2010 Mickle ... 514, 183 - 4-1 pS et al. 7,776,917 B2 * 8/2010 Mickle ...... 514,561 5.534496 A 7/1996 Lee et al. 2001/0026624 A1 10, 2001 Kon et al. 5,594,110 A 1/1997 Fiume et al. 2001/003 1873 A1 10, 2001 Greenwald et al. 5,610,283 A 3/1997 Buechler 2002/0031236 A1 3/2002 Shimizu et al. 5,618,926 A 4, 1997 Salamone et al. 2002/00590 13 A1 5/2002 Rajala et al. 5,643,957 A 7/1997 Leone-Bay et al. 2002fO095134 A1 7, 2002 Pettis et al. 5,670477. A 9, 1997 Poduslo et al. 2002/0097.947 A1 7, 2002 Lim et al. 5,681,811 A 10/1997 Ekwuribe 2002/0098.999 A1 7/2002 Gallop et al. 3.2993. A 2. SA et h 2002/00990 13 A1 7/2002 Piccariello et al. 5741,705. A 4, 1998 East al. 2002/0151526 A1 10/2002 Gallop et al. I k 2002/0151529 A1 10/2002 Cundy et al. 5,756,291. A 5/1998 Griffin et al. 2002/0164373 Al 1 1/2002 Maloney 322999 A 3. E. 1 2002/0173468 A1 11/2002 Lerchen et al. 5,776.885. A 7, 1998 Ei 2002/0183390 A1 12/2002 Javitt w - i. 2003,0004177 A1 1/2003 Kao et al. 3. A 3. SR sal 2003/0077297 A1 4, 2003 Chen et al. 5,811,127 A 9/1998 What al. 2003/009 1593 Al 52003 Bachmann et al. 5,820.881 A 10/1998 Milstein 2003. O130205 A1 7, 2003 Christian 5.843,634 A 2, 1998 fal 2004/0052731 A1 3, 2004 Hirsh et al. 5,846,743 A 2, 1998 Janmeyet al. 2004/0058946 A1 3/2004 Buchwald et al. 5,85536 A 2, 1998 Yageretal 2004/0063628 A1 4/2004 Piccariello et al. 5.861.387 A 1/1999 Labrieet al. 2004/0106636 A1 6/2004 Kream 5,863,899 A 1/1999 Cheronis et al. 2004/O131680 A1 7/2004 Goldenheim et al. 5,882,645 A 3, 1999 Toth et al. 2004/O132968 A1 7/2004 Reed et al. 5,891.459 A 4, 1999 Cooke et al. 2004/O161382 A1 8/2004 Yum et al. 5,891,478 A 4, 1999 Johnson et al. 2004/018003.6 A1 9, 2004 Ashton et al. 5,898,033. A 4, 1999 Swadesh et al. 2004/0204434 A1 10, 2004 Shafer et al. 5,910,569 A 6/1999 Latham et al. 2005, OO38121 A1 2/2005 Mickle et al. US 8,343,927 B2 Page 3

2005.0054561 A1 3/2005 Mickle et al. De Vrueh, Remco, L.A., et al., Transport of L-Valine-Acyclovir via 2005, OO65086 A1 3/2005 Kirk et al. the Oligopeptide Transporter in the Human Intestinal Cell Line, 2005, OO6955O A1 3/2005 Piccariello et al. CACO-2, Journal of Pharmacology and Experimental Therapeutics, 2005, OO74424 A1 4/2005 Salnikov 286(2): 1166-1170 (1988). 2005/OO74425 A1 4/2005 Waugh et al. Deutsche Banc Alex Bornw 2000 Health Care Conference Presenta 2005, 00800 12 A1 4/2005 Mickle et al. 2005, 0112088 A1 5, 2005 Zhao et al. tion, May 10, 2000, Baltimore, MD. 2005/0176644 A1 8, 2005 Mickle et al. Deutsche Banc Alex Brown 2000. 2005/0176645 A1 8, 2005 Mickle et al. Deutsche Banc Alex Brown 2000 Health Care Conference Presenta 2005/0176646 A1 8, 2005 Mickle et al. tion, May 10, 2000, Baltimore, MD. 2005/0266O70 A1 12/2005 Mickle et al. Feb. 10, 2000, Lotus Presentation. 2006, OO14697 A1 1/2006 Mickle et al. Feb. 2000 Presentation to Andrx. 2007/0042.955 A1 2/2007 Mickle et al. Final Report, Study Completion Date Jun. 25, 1998. 2007/00605OO A1 3/2007 Mickle et al. Friedrichsen, G.M., et al., Model Prodrugs Designed for the Intesti 2007/0066537 A1 3/2007 Mickle et al. nal Peptide Transporter a Synthetic Approach for Coupling of 2007/O197451 A1 8, 2007 Mickle et al. Hydroxy-Containing Compounds to Dipeptides Eur J Pharm Sci. 2007/02O3055 A1 8, 2007 Mickle et al. 2008/0090771 A1 4/2008 Moncrief 1491): 13-19 (2001), Abstract. 2009/0234018 A1* 9/2009 Mickle ...... 514,626 Guo, A. et al., Interactions of a Nonpeptidic Drug Valacyclovir, With 2009, 0239949 A1* 9, 2009 Mickle ..... 514,561 the Human Intestinal Peptide Transporter (HPEPT1) Expressed in a 2010, O292336 A1* 11, 2010 Mickle ..... 514,626 Mammalinan Cell Line, Pharmacol Exp Ther, 289(1):448-454 2010, O292337 A1* 11, 2010 Mickle ..... 514,626 (1999) Abstract. 2011/0213034 A1* 9, 2011 Mickle ...... 514,565 Han H.K. et al., Cellular Uptake Mechanism of Amino Acid Ester Prodrugs in CACO-2HPEPT1 Cells Overexpressing a Human FOREIGN PATENT DOCUMENTS Peptide Transporter, Pharm Res, 15(9): 1382-1386 (1998) Abstract. EP O187547 A2 7, 1986 Han, H. et al., 5-Amino Acid Esters of Antiviral Nuclosides, FR 1421130 A 12, 1965 Acyclovir and AZT and Absorbed by the Intestinal PEPT1 Peotide GB 1092089. A 11, 1967 Transporter, Pharm Res. 15(8): 1154-1159 (1998) Abstract. GB 1112347 A 5, 1968 Han, Hyo-Kyung, et al., Targeted Prodrug Design to Optimize Drug WO WO-94.11021 A1 5, 1994 Delivery, AAPS Pharmsci, 2(1):Article 6 (2000). WO WO-95.12605 A1 5, 1995 WO WO-95.14033 A1 5, 1995 Havranova, Marie et al., A High-Molecular Mass Derivative of WO WO-9736616 A2 10, 1997 Trypsin-Kallikrein Inhibitor for Potential Medical Use, II, Hoppe WO WO-98.04277 A1 2/1998 Seyler's Z. Physiol. Chem., 363:295-303 (1982). WO WO-993.9691 A2 8, 1999 Herrera-Ruiz, D., et al., Spatial Expression Patterns of Peptide Trans WO WO-9949.901 A1 10, 1999 porters in the Human and Rate Gastrointestinal Tracts, CACO-2 In WO WO-00371.03 A2 6, 2000 Vitro Cell Culture Model, and Multiple Human Tissues, AAPS WO WO-OO52O78 A1 9, 2000 Pharmsci. 3(1):E9 (2001), Abstract. WO WO-0053233 A1 9, 2000 Hosztafi, S. et al., Synthesis and Analgetic Activity of Nicotine Esters WO WO-O234237 A1 5/2002 of Morphine Derivatives, Arzneim.-Forsch/Drug Res. 43(II), NR. 11 WO WO-03034980 A2 5, 2003 (1993). WO WO-2004/064839 A1 8, 2004 Hughes, et al., Lipidic Peptides, III: Lipidic Amino Acid and WO WO-2004082620 A2 9, 2004 Oligomer Conjugates of Morphine, Journal of Pharmaceutical Sci WO WO-2005.1.18642 A2 12/2005 ences, vol. 80, No. 12, Dec. 1991. WO WO-2006059 106 A2 6, 2006 International Search Report for PCT/US03/05524 Dated Feb. 24, 2003. OTHER PUBLICATIONS International Search Report for PCT/US03/05525 Dated Oct. 9, Apr. 7, 2000, Letter to Astrzeneca LP. 2003. Apr. 7, 2000, Letter to Banc of America Securities, LLC. International Search Report for PCT/US04/17204 Dated Oct. 15, Apr. 7, 2000, Letter to Chase, Hambrecht and Quist. 2004. Apr. 7, 2000, Letter to Credit Suisse First Boston Corporation. International Search Report, Dated Sep. 3, 2003. Apr. 7, 2000, Letter to Johnson & Johnson. Introducing Polythryoid, Presentation, Mar. 2000. Apr. 7, 2000, Letters to Bear, Stearns & Company, Inc. Investment Banking (Long) Presentation, Apr. 27, 2000. Aug. 31, 1999 Presentation to SCIOS, Inc. Investment Banking (Short) Presentation, Apr. 27, 2000. Bai et al., Structural Specificity of Mucosal-Cell Transport and Investment Banking Presentation, Mar. 27-31, 2000. Metabolism of Peptide Drugs; Implication for Oral Peptide Drug Kim et al., B.M. Journal of Pharmaceutical Sciences, vol. 90 (11), p. Delivery. Pharmaceutical Research, Kluwer Academic Publishers, 1767-1775, Nov. 2001. Knutter, I, et al., A Novel Inhibitor of the Mammalian Peptide Trans NY NY, vol. 9, No. 8, Jan. 1, 1992, pp. 969-978, XPO08069566. porter PEPT1, Biochemistry, 40(14):4454-4458 (2001), Abstract. Balimane P.V., et al., Direct Evidence for Peptide Transporter Kovacs, J. et al., Glutamic and Aspartic Anhydrides, Rearrangement (PEPT1)-Mediated Uptake of a Nonpeptide Prodrug, Valacyclovir, ofN-Carboxyglutamic, 1.5-Anhydride to the Leuchs' Anhydride and Biochem Biophys Res Commun, 250(2):246-251 (1998), Abstract. Conversion of the Latter to Pyroglutamic Acid, 85:1839-1844 (Jun. Balimane, P. et al., Effect of Ionization of the Variable Uptake of 20, 1963). Valacyclovir via the Human Intestinal Peptide Transporter (HEPT1) KPMG Auditors' Report, Mar. 12, 1999. In CHP Cells, Biopharm Drug Dispos, 2195): 165-174 (2000), Kramer, Werner et al., Intestinal Absorption of Peptides by Coupling Abstract. to Bile Acids, The Journal of Biochemistry, 269(14) 10621-1627 Bankers Presentation, Mar. 27-31, 2000. (1994). Bunevicius, R. Effects of Thyroxine AS Compared With Thyroxine Kumar, et al., Safety and Pharmacokinetics of Abacavir Following Plus Triodothyronine in Patients With Hypothyrodisim, The New Oral Administration of Escalating Single Doses in Human England Journal of Medicine, vol. 340, No. 6 (1999). Immunodeficiency Virus Type 1-Infected Adults, Antimicrobial Burnette. Thimysta C., et al., Metabolic Disposition of the Acyclovir Agents and Chemotherapy, vol.43, No. 3, Mar. 1999, pp. 603-608, Produg Valacicolovir in the Rat, Drug Metabolism and Disposition, XPOO2530985 ISSN: OO66-4804. 22(1):60-64 (1994). Leibach, F.H. et al., Peptide Transporters in the Intestine and the Canaris G. The Colorado Thyroid Disease Prevalence Study, Kidney, Annu Rev Nutri, 16:99-119 (1996) Abstract. Archives Internal Medicine Articles and Abstracts, vol. 160, No. 4 Leopold, Caludia S. et al., in Vivo Pharmacokinetic Study for the (2000). Assessment of Poly(L-Aspartic Acid) As a Drug Carrier for Colon US 8,343,927 B2 Page 4

Specific Drug Delivery, Journal of Pharmacokinetics and U.S. Appl. No. 10/923,088, Entitled Active Agent Delivery Systems Biopharmaceutics, Vol. 23, No. 4, 1995, p. 397-406, XP002530986. and Methods for Protecting and Administering Active Agents, Li, Chun, et al., Complete Regression of Well-Estabilished Tumors Mickle, et al., filed Aug. 23, 2004. Using a Novel Water-Soluble Poly(L-Gluatmic Acid)-Paclitaxel U.S. Appl. No. 10/953,111, Entitled Compounds and Compositions for the Prevention of Overdose of Oxycodone, Mickle et al., filed Conjugate, Cancer Res, 58:2404-2409 (1998). Aug. 23, 2004. Ma, Y. et al., Enzymatic Mechanism of Thyroxine Biosynthesis, U.S. Appl. No. 1 1/179,801. Entitled Carbohydrate Conjugates to Identification of the Lost Three-Carbon Fragment, J. Am. Chem. Soc. Prevent Abuse of Controlled Substances, Mickle, et al., filed Jul. 13, 1999, vol. 121, No. 38, p. 8697-8698. 2005. Mar. 15, 2000 Presentation to BASF. U.S. Appl. No. 1 1/392,878, Entitled Pharmaceutical Compositions Marriq, Caludine, et al., Amino Acid Sequence of the Unique 3.5.3'- for Prevention of Overdose or Abuse, Mickleet al., filed Apr. 4, 2006. Trilodothyronine-Containing Sequence From Porcine U.S. Appl. No. 1 1/400,304. Entitled Abuse Resistant Amphetamine Thyroglobulin, Biochemical and Biophysical Research-Communi Prodrugs, Mickle et al., filed Apr. 10, 2006. U.S. Appl. No. 11/933,846: Non-Final Office Action Dated Jun. 11, cations, 112(1):206-213 (1983). 2010, Including Form PTO-892 (11 Pages). Negishi, Naoki, et al., Coupling of Naltrexone to Biodegrable Poly Zunino, Franco, et al., Anti-Tumor Activity of Daunorubicin Linked (?-Amino Acids), Pharmaceutical Research, 4(4):305-310 (1987). to Poly-L-Aspartic Acid, International Journal of Cancer, 30:465 Negishi, Naoki, et al., Coupling of Naltrexone to Biodegrable Poly 470 (1994). (O-Amino Acids), Pharmaceutical Research, 4(4):305-310 (1987). Zunino, Franco, et al., Comparison of Antitumor Effects of Oh, D., et al., Estimating the Fraction Dose Absorbed From Suspen Daunorubicin Covalently Linked to Poly-L-Amino Acid Carriers, sions of Poorly Soluble Compounds in Humans: A Mathematical European Journal of Cancer & Clinical Oncology, 20(3):121-125 Model, Pharmaceutical Research, vol. 10, No. 2 (1993). (1984). Oh, DM, et al., Drug Transport and Targeting. Intestinal Transport, Answer and Counterclaim (Feb. 28, 2011) in Shire LLC v. Kempharm Pharma Biotechnol, 12:59-88 (1999) Abstract. (W.D. Va.), Civil Action No. 7:10-CV-00434. Okada, Masahiko, et al., Synthesis of Glycopeptide-Conjugates via Bai, J.P.F. et al., Gastroinestinal Transport of Peptide and Protein Ring-Opening Polymerization of Sugar-Substituted O-Amino Acid Drugs and Prodrugs: In Welling PG, Balant LP, eds. Handbook of N-Carboxyanyhydrides (Glyconcas), Proc. Japan Acad., 73:205-209 Experimental Pharmacology Heidelberg: Springer-Verlag, (1997). 1994:110:189-206. Orten, James Metal. Thyroxine, Human Biochemistry, 9th Ed. C.V. Bennett, D.B., et al.; Drug-coupled Poly(amino Acids) as Polymeric Mosby Company St. Louis, pp. 401-405 (1975). Prodrugs Journal of Bioactive and Compatible Polymers. vol. 3, pp. 44-52 (1988). Pade, V., et al., Link Between Drug Absorption Solubility and Per Bennett, R., et al., “O-Phosphoric Acid Esters of 3,5-Diiodotyrosine meability Measurements in CACO-2Cells, Journal of Pharmceutical and Thyroxine,” Journal of Medicinal and Pharmaceutical Chemis Sciences, vol. 87, No. 12 (1998). try, 2(5): 493-498 (1960). Pharma Presentation, Apr. 2000. Bennett, Raymond, et al., “O-Phosporic Acid Esters of 3,5- Pharma Presentation, Mar. 27-31, 2000, New York, NY. Diiodoyrosine and Thyroxine.” Chemical Abstracts, 55(9):8303 Pharmaceutical Presentation, Mar. 27-31, 2000. (1961). Presentation to Knoll Pharmaceutical, Apr. 10, 2000. Cohen B.M. Journal of Asthma, 1984; V. 21(5) pp. 305-309. Promise of Polythroid, Presentation, Mar. 2000. Franssen et al., “Low Molecular Weight Proteins as Carriers for Rawitch, Allen B., et al., The Isolation of Indentical Thyroxine Con Renal Drug Targeting. Preparation of Drug-Protein Conjugates and taining Amino Acid Sequences From Bovine, Ovine and Porcine Drug-spacer Derivatives and Their Catabolism in Renal Cortex Thyroglobulins, Biochemical and Biophysical Research Communi Homogenates and Lysosomal LyStates.” J. Med. Chem. 1992: 35: cations, 118(2): 423-429 (1984). 1246-1259. Ryser, Hugues J.P. et al., Conjugation of Methotrexate to Poly Furukawa, et al., “Effects of Glu-His-pro-amphetamine (TRH-am (L-Lysine) Increases Drug Transport and Overcomes Drug Resis phetamine) guinea-pig: Antagonistic effect of amphetamine on TRH tance in Cultured Cells, Proc. Natl. AcadSci USA, 75(8):3867-3870 response”, European Journal of Pharmacology, Elsevier BV. NL, (1978). vol. 112, No. 2, Jun. 7, 1985, pp. 237-241, XPO25547293. Sawada, Kyoko, et al. Recognition of L-Amino Acid Ester Com Giammona et al., Coupling of antiviral agent Zidovudine to pounds by Rat Peptide Transporters PEPT1 and PEPT2, Journal of polyapartamide and in vitro drug release studies, Journal of Con Pharmacology and Experimental Therapeutics, 291(2):705-709 trolled Release 1998 vol. 54, pp. 321-331. Greene, Theodora et al., “Protection for Phenols.” Protective Groups (1999). in Organic Synthesis, 2" Ed., John Wiley & Sones, Inc. (1991). Schmidt, Brigitte F., et al., Peptide-Linked 1.30Dialkyl-3- Hussain et al. Synthesis and Structural Eludication of Lipophilic AcyltriaZenes: Gastrin Receptor Directed Antinoeplastic Alkylating Azidothymidine Conjugates. Liebigs Ann. Chem. 1992, pp. 169-171. Agents, Journal of Medicinal Chemistry, 37(22):3812-381791994), Jung et al., Synthesis and In Vitro/In Vivo Evaluation of 1994. 5-Aminosalicyl-Glycine. Journal of Pharmaceutical Sciences May Shen, H. et al., Developmental Expression of PEPT1 or PEPT2 in Rat 2000 vol. 89, No. 5, pp. 594-602. Small Intestine, Colon, and Kidney, Pediatr Res, 49(6):789-795 Kawai, Tohru, et al., “Direct Polymerization of N-Carboxyl Anhy (2001) Abstract. dride of L-Glutamic Acid, Makromol. Chem., 182:2127-2137 Shiraga T., et al., Cellular and Moelcular Mechanisms of Dietary (1981). Regulation of Rat Intestinal H+/Peptide Transporter PEPT1, Gastro Kinoshita et al., Serum Leucine Aminopeptidase Assay . . . Japanese enterology, 116(2):354-362 (1999) Abstract. Journal of Clinical Chemistry 1993, vol. 22, pp. 143-146. Supplementary European Search Report for EP 01273.387 Dated Sep. Kuchimanchi KR, et al., “Intestinal absorption and biodistribution of 28, 2004. cosalane and its amino acid conjugates: novel anti-HIV agents' Inter Supplementary Partial European Search Report for EP 01966056, national Journal of Pharacuetics 231 (2002) 197-221. Dated Apr. 7, 2005. Matsumoto et al. Double-Drugs'—A New Class or Prodrug . Tamai, I., et al., Improvement of L-Dopa Absorption by Dipeptidyl Bioorganic & Medicinal Chemistry Letters, 2000, vol. 10, pp. 1227 Derivation, Utilizing Peptide Transporter PEPT1, J Pharma Sci. 1231. 87(12): 1542-1546 (1998) Abstract. Nariyoshi, Ebihara, et al., “Polyamino Acid Block Copolymer and Toft, A. Thyroid Hormone Replacement—One Hormone or Two?, Preparation Thereof.” (Abstract of JP55145736) (Nov. 13, 1980). The New England Journal of Medicine, vol. 340, No. 6 (1999). Nishida, Koyo, et al., “Pharmacokinetic Analysis of in Vivo Metabo Toth, Isivan, A Novel Chemical Approach to Drug Delivery: Lipidic lism of Amino Acid or Dipetide Conjugates of Salicyclic Acid in Amino Acid Conjugates, Journal of Drug Targeting, 2:217-239 Rabbit Intestinal Microorganisms.” Pharmaceutical Research, 11(1): (1994). 160-164 (1994). US 8,343,927 B2 Page 5

Perisico, F.J., et al., “Effect of Tolimetin Glycine Amide (McN-4366), Smith CB et al., “Dihydromorphine-peptide hybrids have mu recep a Prodrug of Tolimetin Sodium, on Adjuvant Arthritis in the Rate.” tor antagonistic and delta receptor agonistic activity on the mouse The Journal of Pharmacology and Experimental Therapeutics, was deferens and bind with high affinity to opioid receptors in rat 247(3): 889-896 (1986). brain.” NIDA Res Monogr. 75:189-92 (1986). Physician's Desk Reference, "Acuprin 18 Adult Low Dose Aspirin Smith Richard H., et al., “1,3-Dimethyl-3-acyltriazenes: Synthesis Contaqins 81 Mg of Enteric Coated Aspirin.” Jan. 1, 1996, and Chemistry of a Novel Class of Biological Methylating Agents.” XP002932207.2 pages. J. Org. Chem. 51(20):3751-3757 (1986). Portoghese, P.S. et al., J. Med Chem (1997(30(11) pp. 1991-1994. U.S. Appl. No. 12/169,389: Non-Final Office Action dated Dec. 10, Schenk, J., The functioning neuronal transporter for dopamine: 2010, Including Form PTO-892 (23 pages). kinretic mechanisms and effects of amphetamines, cocaine and Ueki, Masaki, et al., Methylphosphinyl (Omp): A New Protecting methyphenidata, Progress in Drug Research, vol. 59, 2002. Group of Tyrosine Suitable for Peptide Synthesis by Use of Boc Schmidt, Briggitte F., et al., “Peptide-Linked 1,3-Dialkyl-3- Amino Acids, I Tetrahedron Letters, 27(35):4181-4184 (1996). acyltriaZenes: Gastrin Receptor Directed Antineoplastic Alkylating Weber, et al., “Synthesis. In Vitro Skin Permaeation Studies, and Agents.” Journal of Medicinal Chemistry, 37(22): 3812-3817 (1994). PLS-Analysis of New Naproxen Derivatives.” Pharmaceutical Shimizu, NS et al., “Inhibition of Infection of T-cell with Human Research, 2001: 18(5):600-607. Immunodefficiency virus type 1 by dideoxynuclosides conjugates U.S. Appl. No. 13/267,585, filed Oct. 6, 2011. with oligopeptides' Antiviral Chemistry and Chemotherapy (1995) 6(1), 17-24. * cited by examiner U.S. Patent Jan. 1, 2013 Sheet 1 of 86 US 8,343,927 B2

FIGURE 1

Yi 1. LiN(TMS), OMF s s 2. Galactose Chloroforate, M. A A 9. h3CO DMF Hsco Yo --O ) o X -A- “HC U.S. Patent Jan. 1, 2013 Sheet 2 of 86 US 8,343,927 B2

p : gi: O CD his

3 is E (O R CN : D C) H O t

O se O

(ufu) uoleuueouoo U.S. Patent Jan. 1, 2013 Sheet 3 of 86 US 8,343,927 B2

FIGURE 3

1. LiNTMS), DMF O 2. Ribose Chloroformate, ( ) O KX l lco DMF HCO O vost OMe d-b

3. MHC

YCH Y

HCO O -so U.S. Patent Jan. 1, 2013 Sheet 4 of 86 US 8,343,927 B2

FIGURE 4

-0-Ribose-HC -O-Hydrocodone 8 O

40 20

O 2O 40 60 80 1 OO 120 Time (minutes) U.S. Patent US 8,343,927 B2 U.S. Patent Jan. 1, 2013 Sheet 6 of 86 US 8,343,927 B2 U.S. Patent Jan. 1, 2013 Sheet 7 of 86 US 8,343,927 B2

s & 3y 8 & 9 (ufu) uoblueouo U.S. Patent Jan. 1, 2013 Sheet 8 of 86 US 8,343,927 B2

FIGURE 1 O -O-Hydrocodone 40 -0-PPL-HC

O O 20 30 40 50 60 Time (minutes post injection)

FIGURE 11

35.0 -O-Hydrocodone --L-HC 3O.O -A-PP-HC 25.0 --GGGGL-HC -0-GGGGL-HC (2X) a 200 T TO; 15.0 3. 10.0 L 5. O

O. O O O.2 0.4 O.6 0.8 Time (hours post injection) U.S. Patent Jan. 1, 2013 Sheet 9 of 86 US 8,343,927 B2

FIGURE 12 2 O 1 8 16 14

-o-Hydrocodone -- GGGGL-HC

O O.2 0.4 0.6 O.8 12 Time (hours post dose)

FIGURE 13

400

350

300 -O-Hydrocodone --EESEEHC --GGGG-HC

250 C-L-HC

200

150

100

O 10 2O 30 40 50 60 Time (minutes) U.S. Patent Jan. 1, 2013 Sheet 10 of 86 US 8,343,927 B2

3 g- O a

1s s SH E 8 E gr

- - - - - O S 3 3 O N N y (ufu) uolajuebuoo U.S. Patent Jan. 1, 2013 Sheet 11 of 86 US 8,343,927 B2

FIGURE 15

700

600

500 -O-Hydrocodone -A-Gal-HC 400

300

200

100

O 10 20 30 40 50 60 Time (minutes) U.S. Patent Jan. 1, 2013 Sheet 12 of 86 US 8,343,927 B2

FIGURE 16

-o-HC 2 5 -0-EEGGl-HC 12 5O 1 O

O 10 20 30 40 50 60 7O Time (Minutes Post IV Dose) U.S. Patent Jan. 1, 2013 Sheet 13 of 86 US 8,343,927 B2

r 3

2 S3 8 C - s

s e o E ?yt E 5 E .

8 & 3 S & 3 8 & 9 & (ufu) uoeueouoco U.S. Patent Jan. 1, 2013 Sheet 14 of 86 US 8,343,927 B2

8 (9

S. g ce

O 3 8 8 & (ufu) uoenueouo U.S. Patent Jan. 1, 2013 Sheet 15 of 86 US 8,343,927 B2

O 3 o O 8 : ? 5, S2

C w

o E S3 E. t E

d N

O y

O C O O. O. O. O. O. O. O. O. O. d o co V. CN C CO to St. CN N V we v y (ufu) uobuluetouoco U.S. Patent Jan. 1, 2013 Sheet 16 of 86 US 8,343,927 B2

3S.

3 O Ot N 3 & 3 S 8 t turfu) uoblueguo U.S. Patent Jan. 1, 2013 Sheet 17 of 86 US 8,343,927 B2

; O: 8

O w

y G)

oc) E

CN E : D t S

O y

3 g vg g gth g g g a (ufu) uoleuuetouoo U.S. Patent Jan. 1, 2013 Sheet 18 of 86 US 8,343,927 B2

C N

2 () 8 8 8 is 9

? S. OY : S O E. C () co E

O N

O y

O O O O O O . O O O CN N se t (uffju) uoeuueouo) U.S. Patent Jan. 1, 2013 Sheet 19 of 86 US 8,343,927 B2

8 3 8 s & 3 & 8 S. 8 t (ufu) uoeueoudo U.S. Patent Jan. 1, 2013 Sheet 20 of 86 US 8,343,927 B2

O O

C C

9 O 9.

C. w

C C C C C C C C C C, C r N C C D N N N V re - Y - (ufu) uoeueouoso U.S. Patent Jan. 1, 2013 Sheet 21 of 86 US 8,343,927 B2

O O L O C N N y re (u/6u) uoleuueouoco U.S. Patent Jan. 1, 2013 Sheet 22 of 86 US 8,343,927 B2

S3 R U

t S.

8 S. E. E t

O & 8 S. 3 & O U.S. Patent Jan. 1, 2013 Sheet 23 of 86 US 8,343,927 B2

3 O 8 y ty (uffu) uoeulueouoo U.S. Patent Jan. 1, 2013 Sheet 24 of 86 US 8,343,927 B2

()

5 S O C O t L

O c

E OO o 5 CN c E O m CD H T

O N

C y

O O O C c O C O O N CN y vis U.S. Patent Jan. 1, 2013 Sheet 25 of 86 US 8,343,927 B2

d O ; : S2 NZst 9 9 O w

o 5 c) O E CN P r A. D O t N

O s

O O O C O O C L N tu?su)t uoeueouo)y U.S. Patent Jan. 1, 2013 Sheet 26 of 86 US 8,343,927 B2

FIGURE 3 O

200 -O-Hydrocodone -A-DDFF-HC 80 160 e 140 S. S 120 5 100 80 2 8 60 40 20 O O 10 2O 30 40 50 60 Time (minutes)

FIGURE 31

250 -O-Hydrocodone -A-YYEE-HC 200 E S 150 9.

u? OO C O O 50

O 10 20 30 40 50 60 Time (min) U.S. Patent Jan. 1, 2013 Sheet 27 of 86 US 8,343,927 B2

FIGURE 32 250 -O-Hydrocodone

E g 150 9. 100 9 O () 50

O O O 20 30 40 50 60 Time (minutes)

FIGURE 33 250 -o-Hydrocodone --KKEE-HC 200 O -A-EEFF-HC

S-150 O 100

O 50 -

O O O.2 0.4 O6 0.8 1 Time (hours) U.S. Patent Jan. 1, 2013 Sheet 28 of 86 US 8,343,927 B2

FIGURE 34.

18O -O-Hydrocodone --YYFP-HC

O O

O O2 0.4 O.6 0.8 Time (hours)

FIGURE 35

cro O Y 20%.rise Phosgene D."...a? X O JX in toluene O O -\ 6 -A-6 U.S. Patent Jan. 1, 2013 Sheet 29 of 86 US 8,343,927 B2

FIGURE 36 -O-hydrocodone 200 rGa-P2-HC 18O --Ga-P2L-HC

160 14 O 20 OO : O O 0.2 0.4 O.6 0.8 Time (hours)

FIGURE 37

-o-Hydrocodone 4 O -O-Gu-l-HC 23 OO

1 O

O 0.5 1.5 2 2.5 3 Time (hours) U.S. Patent Jan. 1, 2013 Sheet 30 of 86 US 8,343,927 B2

FIGURE 38

O NH2 NH2 O

HO- O1 N HO- O19. N HO KlNYN HO “l.NYN NH

HO HO HO HO Thymidine 2'-deoxycytidine 2'-deoxyadenosine 2'-deoxyguanosine Representative Nucleosides s O TEBDMSO \oyon HO -- al Conjugation Site Protected Thymidine Site of Conjugation for Hydrocodone

4. FIGURE 39 2

1 O --Hydrocodone as EEFF-HC U.S. Patent Jan. 1, 2013 Sheet 31 of 86 US 8,343,927 B2

1 2 - FIGURE 4 O

1 O d

-0-Hydrocodone

6 --EEFFF-HC

2 -

O O 2 3 4 5 6 7 8 hours

FIGURE 41 1O 9 8 is 7 a0HC 6 -HYY-HC E 5 5, 4 3 2 1 O O 2 4. 6 8 Hours U.S. Patent Jan. 1, 2013 Sheet 32 of 86 US 8,343,927 B2

18 FIGURE 42 16

14 -0HC

12 DD-HC 10 U.S. Patent Jan. 1, 2013 Sheet 33 of 86 US 8,343,927 B2

FIGURE 43

O U.S. Patent Jan. 1, 2013 Sheet 34 of 86 US 8,343,927 B2

FIGURE 44 100 90 80 -o-Hydrocodone 70 mEEFF-HC

U.S. Patent Jan. 1, 2013 Sheet 35 of 86 US 8,343,927 B2

FIGURE 45

O 2 4 6 8 Hours

FIGURE 46 120

80 a0HC

60 a DD-HC E Sh U.S. Patent Jan. 1, 2013 Sheet 36 of 86 US 8,343,927 B2

80 FIGURE 47

70 s 60 -0-HC 50 -s-YYFF-HC E 40 5. 30 20 10 O O 1 2 3 4. 5 6 7 8 Hours

FIGURE 48

300

250

2 O O -0-Hydrocodone an EEFFF-HC OO 5 O

O O 10 2O 30 40 50 60 Minutes U.S. Patent Jan. 1, 2013 Sheet 37 of 86 US 8,343,927 B2

FIGURE 49 3OO

250 HC a 200 --- YY-HC

D 15O S OO

O O O 2O 3O 40 5O 60 Minutes

FIGURE 5 O

600

SOO

s 400 DD-HC 2 3OO E ? 200

O O 10 2O 30 40 50 SO

Minutes U.S. Patent Jan. 1, 2013 Sheet 38 of 86 US 8,343,927 B2

FIGURE 51. 600

500 --HC 400 -HYYFF-HC

2OO

O 10 2O 30 40 50 60

Minutes

FIGURE 52

250 -0-Hydrocodone on EEFF-HC 200 150 100 50

O 1 O 2O 3O 40 5O 6O Minutes U.S. Patent Jan. 1, 2013 Sheet 39 of 86 US 8,343,927 B2

FIGURE 53

250 -Hydrocodone of EEFFF-HC 2OO s g s 15O

sS 100

Minutes

FIGURE 54

6OO

5OO

-- YY-HC r 300 E 20o 1 OO

O O 1 O 2O 3O 40 50 6O

Minutes U.S. Patent Jan. 1, 2013 Sheet 40 of 86 US 8,343,927 B2

FIGURE 55

-HYYFF-HC

O 10 20 30 40 50 60 Minutes

FIGURE 56 140

8 O -O-HC1 mg/kg

6 O -0-YYFF-HC 1 mg/kg

4. O

2 O

O 0.0 0.5 1.0 5 2.0 2.5 3.0 3.5 4.0 Time (hours) U.S. Patent Jan. 1, 2013 Sheet 41 of 86 US 8,343,927 B2

120 FIGURE 57

100 -O-HC 1 mg/kg 8 O -0-YYFF-HC1 mg/kg

4.6OO

2 O

0.0 0.5 1.0 15 2.0 2.5 3.0 3.5 4.0 Time (hours) U.S. Patent Jan. 1, 2013 Sheet 42 of 86 US 8,343,927 B2

FIGURE 58

-o-HC 1 mg/kg --YYFFI-HC 1 mg/kg

0. 5

O 0.0 0.5 O 15 2.0 2.5 3.0 3.5 4.0 Time (hours)

BO FIGURE 59

50 -O-HC 2 mg/kg

-0-YYFFI-HC 2 mg/kg

O.O O.5 O 1.5 2.0 2.5 3.O 3.5 4.O Time (hours) U.S. Patent Jan. 1, 2013 Sheet 43 of 86 US 8,343,927 B2

70 FIGURE 6 O 60

50 -o-HC 2 mg/kg -0-YYFF-HC 2 mg/kg 40

2 O

1 O

0.0 O.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 Time (hours) U.S. Patent Jan. 1, 2013 Sheet 44 of 86 US 8,343,927 B2

16 FIGURE 61 14 -o-HC 5 mg/kg 12 -0-YYFF-HC 5 mg/kg O 8

O.O O 2.0 3.0 4.0 Time (hours) U.S. Patent Jan. 1, 2013 Sheet 45 of 86 US 8,343,927 B2

7 O O FIGURE 62 6 O O

5 O O 400 -O-HC 5 mg/kg 300 -0-YYFF-HC 5 mg/kg

200

100

O.O 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 Time (hours)

700 FIGURE 63 600 E 500 -O-HC 5 mg/kg 4OO -o-YYFF-HC 5 mg/kg

300 o O Cs 2OO

1 OO

O.O 1.0 2.O 3.O 4.O Time (hours) U.S. Patent Jan. 1, 2013 Sheet 46 of 86 US 8,343,927 B2

700 FIGURE 64

6OO 500 -O-HC 5 mg/kg -0-YYFF-HC 5 mg/kg 400

2OO

1 OO

O.O. 1.0 2.0 3.0 4.O Time (hours)

FIGURE 65

5OO 45O 4OO --HC 25 mg/kg 350 -O-YYFF-HC 25

250 - 200 15O OO

O.O O.5 1.O 1.5 2.O 2.5 3.0 3.5 4-0 Time (hours) U.S. Patent Jan. 1, 2013 Sheet 47 of 86 US 8,343,927 B2

45O 4OO N FIGURE 66 S 350 g 3OO S. 2 -O-HC 25 mg/kg 250 -o-YYFFI-HC 25 mg/kg 200 5 150 C) 100 SO O O. O. 1.O. 2.O 3.O 4.O Time (hours)

FIGURE 67

60 -O-HC 25 mg/kg 50 -0-YYFF-HC 25 mg/kg

9 40 s 30 O O 20 s 1 O.

O O.O O.5 1.O 1.5 2.O 2.5 3.0 3.5 4.O Time (hours) U.S. Patent Jan. 1, 2013 Sheet 48 of 86 US 8,343,927 B2

FIGURE 68

-o-Hydrocodone 500 bitantrate -O-YYFF-HC S 4oo c S. 300 us e 2OO C) 3. 1OO O O 5 1O 15 2O 25 Dose (mg/kg)

FIGURE 69

600 -O-Hydrocodone bitantrate 500 -- YYFF-HC S 400 O) S. 300 s cb 200 O D c 100

O 100 200 300 400 Human Equivalent Dose (mg hydrocodone bitantrate) U.S. Patent Jan. 1, 2013 Sheet 49 of 86 US 8,343,927 B2

FIGURE 7 O 700

6OO 5OO 5, 400 3oo -O-Hydrocodone bitantrate -0-YYFF-HC 9 200 100

O O 5 10 15 2O 25 Dose (mg/kg)

FIGURE AL

7OO

6OO

500 E 400 g -O-Hydrocodone bitantrate 300 -0-YYFF-HC O 200

1 OO

O 50 OO 150 2OO 250 3OO 350 400 Human Equivalent Dose (mg hydrocodone bitantrate) U.S. Patent Jan. 1, 2013 Sheet 50 of 86 US 8,343,927 B2

3.O O

FIGURE 72 2 5 O

2 O O -O-HC 1 mg/kg IV OO -O-YYFF-HC 1 mg/kg IV

5 O

C O.O O.S O 15 2.O 2.5 3O 3.5 4-O Time (hours)

FIGURE 73 3 O O

2 5 O

2 O O -0-HC 1 mg/kg IV -o-YYFF-HC 1 mg/kg IV OO

5. O

O O.O O 2.O 3.O 4.O Time (hours) U.S. Patent Jan. 1, 2013 Sheet 51 of 86 US 8,343,927 B2

FIGURE 74

2 5 -0-HC 1 mg/kg IV -O-YYFF-HC 1 mg/kg IV 2 O

1 5

O O.O O.5 1.O 15 2.O 2.5 3.0 3.5 4.O. Time (hours)

FIGURE 75

5OO --HC 1 mg/kg IN

-0-YYFF-HC 1 mg/kg IN 4OO

3OO

2OO

1 OO

O 1 O 2O 30 40 50 6O Time (minutes) U.S. Patent Jan. 1, 2013 Sheet 52 of 86 US 8,343,927 B2

FIGURE 76 6OO

E 5OO G 4OO -O-HC 1 mg/kg IN 5 300 -0-YYFF-HC 1 mg/kg N 200 C 3 100

O 10 20 30 40 50 60 Time (minutes)

FIGURE 77

1 -o-HC 1 mg/kg IN -0-YYFF-HC1 mg/kg IN

O 10 20 30 40 50 60 Time (minutes) U.S. Patent Jan. 1, 2013 Sheet 53 of 86 US 8,343,927 B2

140 FIGURE 78 120

-0-YYFF-HC 1 mg/kg

O.O 0.5 1.O 1.5 2.O 2.5 3.0 3.5 4.O Time (hours)

FIGURE 79 12O.

1OO -O-HC 1 mg/kg 8 O -0-YYFF-HC 1 mg/kg

4.6OO

2 O

O.O. 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 Time (hours) U.S. Patent Jan. 1, 2013 Sheet 54 of 86 US 8,343,927 B2

FIGURE 8 O 3. 5

da 3 -O-HC 1 mg/kg 5 -0-YYFF-HC 1 mg/kg S.2 2.5 C 9, 2 s 1.5 d 3 1 s 0.5

O OO O.5 1.O 1.5 2O 2.5 3.O 3.5 4.O Time (hours)

FIGURE 81 8O 70

60 50 -O-HC 2 mg/kg

40 -0-YYFF-HC 2 mg/kg

O.O O.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 Time (hours) U.S. Patent Jan. 1, 2013 Sheet 55 of 86 US 8,343,927 B2

70 FIGURE 82 60

50 -O-HC 2 mg/kg -0-YYFFI-HC 2 mg/kg

O.O O.5 1.O 1.5 2.O 2.5 3.0 3.5 4.O Time (hours)

FIGURE 83 16 14 -O-HC 5 mg/kg 12 -0-YYFF-HC 5 mg/kg 10

O.O 1.0 2.0 3.0 4.0 Time (hours) U.S. Patent Jan. 1, 2013 Sheet 56 of 86 US 8,343,927 B2

7OO

SOO FIGURE 84 5OO 4OO -O-HC 5 mg/kg 3OO -0-YYFF-HC 5 mg/kg

2OO

O.O O.5 1.O 1.5 2.0 2.5 3.O 3.5 4.0 Time (hours)

7OO FIGURE 85

500 -O-HC 5 mg/kg 400 -0-YYFF-HC 5 mg/kg

300

200

100

O.O 1.0 2.O 3.0 4.0 Time (hours) U.S. Patent Jan. 1, 2013 Sheet 57 of 86 US 8,343,927 B2

7OO FIGURE 86 600 500 -o-HC 5 mg/kg -0-YYFF-HC 5 mg/kg 400

2OO :

O.O 1.O 2.0 3.0 4.O Time (hours)

FIGURE 87 500 450 4OO -O-HC 25 mg/kg 350 -O-YYFF-HC 25 3OO mg/kg 250 200 150 OO 50

O.O O.5 1.O 1.5 2.0 2.5 3.0 3.5 4.O Time (hours) U.S. Patent Jan. 1, 2013 Sheet 58 of 86 US 8,343,927 B2

450 400 2S FIGURE 88 S 350 o 5- 300 5 250 -O-HC 25 mg/kg s -0-YYFF-HC 25 mg/kg E 200 d 32 150 100 50 O O.O 1.0 2.O 3.0 4.0 Time (hours)

FIGURE 89

60 -O-HC 25 mg/kg 50 -0-YYFF-HC 25 mg/kg

2 40 s 5 30 o O 20 S C 10

O.O 0.5 1.O 1.5 2.0 2.5 3.O 3.5 4.O Time (hours) U.S. Patent Jan. 1, 2013 Sheet 59 of 86 US 8,343,927 B2

FIGURE 90 6OO -O-Hydrocodone 500 bitantrate -- YYFF-HC 4OO

3OO

2OO

O O 5 O 15 2O 25 Dose (mg/kg)

FIGURE 91

600 -O-Hydrocodone bitantrate 500 -- YYFF-HC 400

3OO

2OO

100

O 1 OO 2OO 3OO 400 Human Equivalent Dose (mg hydrocodone bitantrate) U.S. Patent Jan. 1, 2013 Sheet 60 of 86 US 8,343,927 B2

700 FIGURE 92

600

500

E 400 300 -o-Hydrocodone bitantrate -0-YYFF-HC O 200

100

O O 5 O 5 2O 25 Dose (mg/kg)

FIGURE 93

700

600

500 S 400 g -o-Hydrocodone bitantrate 300 -0-YYFF-HC O 200

100

O 50 100 150 2OO 250 300 350 400 Human Equivalent Dose (mg hydrocodone bitantrate) U.S. Patent Jan. 1, 2013 Sheet 61 of 86 US 8,343,927 B2

3 O O FIGURE 94 2 5 O

2 O O --HC 1 mg/kg IV -0-YYFF-HC 1 mg/kg IV 5151 OOO O

O.O O.5 1.O 5 2.0 2.5 3.0 3.5 4.0 Time (hours)

FIGURE 95

3.O O

2 5 O

2 O O -o-HC 1 mg/kg IV -o-YYFF-HC 1 mg/kg IV OO

5 O

Time (hours) U.S. Patent Jan. 1, 2013 Sheet 62 of 86 US 8,343,927 B2

35 FIGURE 96

30 25 -0-HC 1 mg/kg IV -o-YYFF-HC 1 mg/kg IV

O.O 0.5 1.O 1.5 2.0 2.5 3.0 3.5 4.O Time (hours)

FIGURE 97

50 O -O-HC 1 mg/kg IN -0-YYFFI-HC 1 mg/kg IN 4 O O

300

200

1 OO

O O 2O 30 40 5O 6O Time (minutes) U.S. Patent Jan. 1, 2013 Sheet 63 of 86 US 8,343,927 B2

6 O O FIGURE 98

5 O O

4. O O -O-HC 1 mg/kg IN

3 O O -0-YYFF-HC 1 mg/kg N 2OO

1 OO

O O 2O 3O 4O 5O 6O Time (minutes)

FIGURE 99

18 16 E Sh 14

S.s 12 10 8 -O-HC1 mg/kg IN

E 6 -0-YYFF-HC 1 mg/kg IN 4 8 2 O O 1O 2O 3O 40 50 60 Time (minutes) U.S. Patent Jan. 1, 2013 Sheet 64 of 86 US 8,343,927 B2

FIGURE 1 OO

H3C n N OH -- Tertiary Ester

H3CO O O - Ester

FIGURE 1 O1 U.S. Patent Jan. 1, 2013 Sheet 65 of 86 US 8,343,927 B2

FIGURE 102

OH HO OH N N OH O O

MeO O O O HO HO OH OH

FIGURE 103

U.S. Patent Jan. 1, 2013 Sheet 66 of 86 US 8,343,927 B2

FIGURE 1 O4

N OH HN- NH 1. OH N S ~~ Niacin Biotin O

FIGURE 105

O 0.5 1. 5 2 2.5 3. time (hours) U.S. Patent Jan. 1, 2013 Sheet 67 of 86 US 8,343,927 B2

FIGURE 1 O6 -O-Oxycodone 90 --- (A)2OC -- (V)2-OC --(Y'YV)2-0C

O O.S 1 .5 2 2.5 Time (hours)

FIGURE Of

12O

O O.5. 5 2 2.5 3 Time (hours) U.S. Patent Jan. 1, 2013 Sheet 68 of 86 US 8,343,927 B2

FIGURE 1 O8

-o-, Oxycodone -- (KKV-OC

O O.S S 2 2.5 3 Time (hours)

FIGURE 1 O9

U.S. Patent Jan. 1, 2013 Sheet 69 of 86 US 8,343,927 B2

FIGURE 11 O

O 0.5 1.5 2 2.5 3. Time (hours

FIGURE 111

-o-Oxycodone --(YYhF)2-CC

line (hours) U.S. Patent Jan. 1, 2013 Sheet 70 of 86 US 8,343,927 B2

FIGURE 112

FIGURE 113

0.5 5. 2 25 3. Time (hours) U.S. Patent Jan. 1, 2013 Sheet 71 of 86 US 8,343,927 B2

FIGURE 114 35 3O -ou Oxycodore 5,a- 25 --(YW2YWY2-OC E. 2 s 15 . 3 to o S -

0.5 1. 15 2 2.5 3 Time (hours)

FIGURE 5

40 --Oxycodone

35 ---(LYW2-OC

Tirne (hours) U.S. Patent Jan. 1, 2013 Sheet 72 of 86 US 8,343,927 B2

FIGURE 116

Time (hours)

FIGURE 117

O O.S 1 15 2 2.5 3. Time (hours) U.S. Patent Jan. 1, 2013 Sheet 73 of 86 US 8,343,927 B2

FIGURE 118 aO 35 -O-Oxycodone E 30 -O-(LYW2-OC G S 25 is5 20 3. 5 5 10 5. O O 0.5 1.5 2 2.5 3. 3.5 Time (hours)

FIGURE 119 40 35 3) 2 5 O201 5

O 0.5 1.5 2 2.5 3 Time (hours U.S. Patent Jan. 1, 2013 Sheet 74 of 86 US 8,343,927 B2

FIGURE 12 O SO -On-Oxycodone e 50 -a- (VYW)2-OC 340 al S 3D 2O

5 O.

OOOO O 5 1S 2 2.5 3. time (hours

FIGURE 121

AO -0- Oxycodone 35 -- (LYV)2-OC(LYV) E 30 - 25 5 20 E 15 3. 8 O 5 O O.S t S 2 2.5 3 fine (hours) U.S. Patent Jan. 1, 2013 Sheet 75 of 86 US 8,343,927 B2

FIGURE 122

b E. O e s 8 o 3

- os 5 2 25 3. Time (hours)

FIGURE 123

60 -o-OxyCodore 50 -- (WYV)2-OC E 40 R 30 as 20 5 9 10

O O O.5 1.5 2 2.5 3. fine (hours) U.S. Patent Jan. 1, 2013 Sheet 76 of 86 US 8,343,927 B2

FIGURE 124

1 O O

5 O

C) O O 2. 3) 40 SO 50 Time (minutes)

FIGURE 125

-o-Oxycodore 350 -O-(FFV)2-OC 300 s --(GG)-OC E 250 --(PP)-Oc s 200 150 S.5 100 O 50 - O La C Sa-F Y were O 2 (0.4 0.6 O.8 Time (hours) U.S. Patent Jan. 1, 2013 Sheet 77 of 86 US 8,343,927 B2

O) FIGURE 126 -- Oxycodone 8OO) -- (EEM2-0C 700 re- (TV)2-OC 600 --(YYV)2-0C g A. -- (PPV)2-0C 5

2 c

0.6 8. Tine (hours)

FIGURE 127 250 -o-Oxycodone 2OO -- (YYW)2-OC E. -- (PPV)2-OC 150 S. 1 OO

O5 50

O O O.2 0.4 O.S 0.8 Tine (hours) U.S. Patent Jan. 1, 2013 Sheet 78 of 86 US 8,343,927 B2

FIGURE 128

BOO -o-Oxycodone

-a (KKW)2-OC g 500 s s 400 300 3 200 r O O O O), 2 O-4 O6 0.8 1 Time (hours)

FIGURE 129 -o-oxycodone 700 -A-D-dB)2-0C -0-(YYKYY2-OC 60 -o-(FFKFF)2-OC

SOO -- (FFCha)2-OC

40 U.S. Patent Jan. 1, 2013 Sheet 79 of 86 US 8,343,927 B2

FIGURE 13 O

700 -o- Oxycodone 600 -a- (PPKPP)2-OC E g 500 -(FFNie)2-oc 5 400 300 200 O P 100 O O O.2 0.4 O6 8 Time (hours)

FIGURE 131 OO -0-a- Oxycodone -- YYNie)2-CC 600 -- (PPCha)2-CC -- (YYCha)-OC 500 -- (YYhf2-OC 400 -o- (Y22 5 is 300 / 200 s - (2-S O Kae O O O.2 O.4 O.S O.8 Time (hours) U.S. Patent Jan. 1, 2013 Sheet 80 of 86 US 8,343,927 B2

FIGURE 132 70

SOO

400

200

Time (hours)

FIGURE 133

-O-Oxycodone

re- (GYW2-OC

-- (YV)2-OC

2 ,4 O. 0.8 f Time hours) U.S. Patent Jan. 1, 2013 Sheet 81 of 86 US 8,343,927 B2

FIGURE 134 -o-OxyCodone 8OO -A- (LYW)2-OC 700 -O-(FYV)2-OC

4. O O

200 OO

O O.2 O4 O.S O.8 Time (hours)

FIGURE 135

800

7. O)

0.2 O.4 0.6 O.B. Time (hours) U.S. Patent Jan. 1, 2013 Sheet 82 of 86 US 8,343,927 B2

FIGURE 136 -o-Oxycodone

O O.2 O-4 .6 G.8 Time (hours)

FIGURE 137

O 3.2 0.4 0.6 0.8 Time (hours) U.S. Patent Jan. 1, 2013 Sheet 83 of 86 US 8,343,927 B2

BOO FIGURE 138 o-Oxycodore

5 O O 24.3. OOO OOO OO

O O.2 O4 O6 O-8 Time (hours)

FIGURE 139

O D2 O.4 0.6 0.8 Time (hours) U.S. Patent Jan. 1, 2013 Sheet 84 of 86 US 8,343,927 B2

900 FIGURE 14 O -O-Oxycodone BOO . --(KFV)2-OC 70 -- (PFW)2-OC 2 SOD 5 500 400 . E 8 300 8 200 OO O a O O2 O4 O.S O.8 Time (hours)

FIGURE 141 -O-Oxycodone 900 -0-(VFW)2-OC 80 . --(FOV)2-OC

O 0.2 4. 0.5 8 t Time (hours) U.S. Patent Jan. 1, 2013 Sheet 85 of 86 US 8,343,927 B2

FIGURE 142 16O -0-Oxycodone --P2L(2)-Oxycodone 1 O O

FIGURE 143

-0-Oxycodone

--P2L(2)-Oxycodone

Hours U.S. Patent Jan. 1, 2013 Sheet 86 of 86 US 8,343,927 B2

FIGURE 144 120 --Oxycodone

100 -HP2L(2)-OxyCOdone

Minutes US 8,343,927 B2 1. 2 PHARMACEUTICAL COMPOSITIONS FOR tial for overdose or abuse is reduced due to the limited bio PREVENTION OF OVERDOSE ORABUSE availability of the active agent as compared to the active agent delivered as free drug. CROSS REFERENCE TO RELATED APPLICATIONS BRIEF DESCRIPTION OF THE FIGURES This application is a continuation of U.S. patent applica FIG. 1. illustrates preparation of Galacto-Hydrocodone. tion Ser. No. 11/089,056, filed Mar. 25, 2005, which is a FIG.2. Oral bioavailability of abuse-resistant hydrocodone continuation-in-part application and claims priority to 35 carbohydrate conjugates, measured as free hydrocodone U.S.C. S 120 to U.S. application Ser. No. 09/933,708 filed 10 (with measured plasma levels by ELISA). Aug. 22, 2001; U.S. patent application Ser. No. 11/089,056 is FIG. 3. illustrates preparation of Ribo-Hydrocodone. also a continuation-in-part and claims benefit under 35 U.S.C. FIG. 4. Intranasal bioavailability of abuse-resistant hydro 120 to U.S. application Ser. No. 10/156.527 filed May 29, codone carbohydrate conjugate, measured as free hydroc 2002, which claims the benefit under 35 U.S.C. 119(e) to U.S. 15 odone (with measured plasma levels by ELISA). Provisional application No. 60/358,368 filed Feb. 22, 2002, FIG. 5. illustrates preparation of Leu-Hydrocodone. and U.S. Provisional application No. 60/366,258 filed Mar. FIG. 6. illustrates preparation of Ala-Pro-Hydrocodone. 22, 2002 and U.S. Provisional No. 60/358,381 filed Feb. 22, FIG. 7. illustrates the preparation of Gly-Gly-Leu-Hydro 2002; U.S. patent application Ser. No. 11/089,056 is also a codone. continuation-in-part and claims benefit under 35 U.S.C. 120 FIG. 8. illustrates preparation of Gly-Gly-Gly-Gly-Leu to U.S. application Ser. 10/953,119 filed Sep. 30, 2004, which Hydrocodone. claims benefit under 35 U.S.C. 119(e) to U.S. Provisional FIG.9. Intranasal bioavailability of abuse-resistant hydro application No. 60/567,800 filed May 5, 2004; U.S. Provi codone amino acid, di- and tri-peptide conjugates, measured sional application No. 60/507,012 filed Sep. 30, 2003: U.S. as free hydrocodone. Provisional application No. 60/567,802 filed May 5, 2004; 25 FIG. 10. Analgesic effect of abuse-resistant hydrocodone and U.S. Provisional application No. 60/568,011 filed on tri-peptide conjugate following intranasal administration, May 5, 2004; U.S. patent application Ser. No. 11/089,056 is measured as free hydrocodone. also a continuation-in-part and claims benefit under 35 U.S.C. FIG. 11. Analgesic effect of abuse-resistant hydrocodone S120 to U.S. application Ser. No. 10/953,110 filed Sep. 30, tri- and penta-peptide conjugates following Subcutaneous 2004; U.S. patent application Ser. No. 11/089,056 is also a 30 administration, measured as free hydrocodone. continuation-in-part and claims benefit under 35 U.S.C. S 120 FIG. 12. Analgesic effect of abuse-resistant hydrocodone to Ser. No. 10/923,257 filed Aug. 23, 2004; U.S. patent appli penta-peptide conjugate following intranasal administration, cation Ser. No. 1 1/089,056 is also a continuation-in-part and measured as free hydrocodone. claims benefit under 35 U.S.C. S 120 to Ser. No. 10/923,088 35 FIG. 13. Intranasal bioavailability of abuse-resistant filed Aug. 23, 2004 which claims benefit under 35 U.S.C. hydrocodone tri- and penta-peptide conjugates, measured as S120 to and is a continuation-in-part application of PCT free hydrocodone. application No. US03/05525 filed Feb. 24, 2003 which claims FIG. 14. Intranasal bioavailability of abuse-resistant benefit under 35 U.S.C. S 119(e) to U.S. Provisional Applica hydrocodone tri- and penta-peptide conjugates, measured as tion No. 60/362,082 filed Mar. 7, 2002; U.S. patent applica 40 free hydrocodone. tion Ser. No. 11/089,056 is also a continuation-in-part and FIG. 15. Intranasal bioavailability of abuse-resistant claims benefit under 35 U.S.C. S 120 to U.S. application Ser. hydrocodone an amino acid-carbohydrate peptide conjugate, No. 10/955,006 filed Sep. 30, 2004; U.S. patent application measured as free hydrocodone. Ser. No. 11/089,056 is also a continuation-in-part and claims FIG. 16. Analgesic effect of abuse-resistant hydrocodone benefit under 35 U.S.C. S 120 to U.S. application Ser. No. 45 penta-peptide conjugate following intravenous administra 10/953,116 filed Sep. 30, 2004; U.S. patent application Ser. tion, measured as free hydrocodone. No. 11/089,056 is also a continuation-in-part and claims ben FIG. 17. Intranasal bioavailability of an abuse-resistant efit under 35 U.S.C. 120 to U.S. application Ser. No. 10/953, hydrocodone tri-peptide conjugate, measured as free hydro 111 filed Sep. 30, 2004; U.S. patent application Ser. No. codone. 11/089,056 is a continuation-in-part and claims benefit under 50 FIG. 18. Intranasal bioavailability of an abuse-resistant 35 U.S.C. S 119 to PCT/US04/32131 filed Sep. 30, 2004. hydrocodone penta-peptide conjugate, measured as free Each of the above applications are hereby incorporated by hydrocodone. FIG. 19. Intranasal bioavailability of an abuse-resistant reference in their entirety. hydrocodone tri-peptide conjugate, measured as free hydro 55 codone. BACKGROUND FIG. 20. Intranasal bioavailability of abuse-resistant hydrocodone tri- and penta-peptide conjugates, measured as Accidental and intentional overdose with prescription and free hydrocodone. over the counter drugs is a serious health problem with thou FIG. 21. Intranasal bioavailability of abuse-resistant sands of fatalities occurring each year as a result. The present 60 hydrocodone penta-peptide conjugates, measured as free invention relates to pharmaceutical compositions comprised hydrocodone. of a chemical moiety attached to an active agent in a manner FIG. 22. Intranasal bioavailability of an abuse-resistant that Substantially decreases the potential of the active agent to hydrocodone penta-peptide conjugate, measured as free cause overdose or to be abused. When delivered at the proper hydrocodone. dosage the pharmaceutical composition provides therapeutic 65 FIG. 23. Intravenous bioavailability of an abuse-resistant activity similar to that of the parent active agent. However, hydrocodone tri-peptide conjugate, measured as free hydro when the composition is delivered at higher doses the poten codone. US 8,343,927 B2 3 4 FIG. 24. Intranasal bioavailability of an abuse-resistant FIG. 47. Oral bioavailability in rats for hydrocodone vs. hydrocodone tri-peptide conjugate, measured as free hydro YYFFI-HC at a dose (5 mg/kg) approaching a human over codone. dose equivalent measured as free hydrocodone. FIG. 25. Oral bioavailability of an abuse-resistant hydroc FIG.48. Decrease in bioavailability ofEEFFF-HC as com odone penta-peptide conjugate, measured as free hydroc pared to hydrocodone by the intranasal route of administra odone. tion measured as free hydrocodone. FIG. 26. Intranasal bioavailability of an abuse-resistant FIG. 49. Decrease in bioavailability of YYI-HC as com hydrocodone tri-penta-peptide conjugate, measured as free pared to hydrocodone by the intranasal route of administra hydrocodone. tion measured as free hydrocodone. 10 FIG. 50. Decrease in bioavailability of DDI-HC as com FIG. 27. Intranasal bioavailability of an abuse-resistant pared to hydrocodone by the intranasal route of administra hydrocodone penta-peptide conjugate, measured as free tion measured as free hydrocodone. hydrocodone. FIG.51. Decrease in bioavailability ofYYFFI-HC as com FIG. 28. Intranasal bioavailability of abuse-resistant pared to hydrocodone by the intranasal route of administra hydrocodone penta-peptide conjugates, measured as free 15 tion measured as free hydrocodone. hydrocodone. FIG.52. Decrease in bioavailability of EEFFI-HC as com FIG. 29. Intranasal bioavailability of an abuse-resistant pared to hydrocodone by the intravenous route of administra hydrocodone tri-peptide conjugate containing D- and L-iso tion measured as free hydrocodone. mers, measured as free hydrocodone. FIG.53. Decrease in bioavailability ofEEFFF-HC as com FIG. 30. Intranasal bioavailability of an abuse-resistant pared to hydrocodone by the intravenous route of administra hydrocodone penta-peptide conjugate, measured as free tion measured as free hydrocodone. hydrocodone. FIG. 54. Decrease in bioavailability of YYI-HC as com FIG. 31. Intranasal bioavailability of an abuse-resistant pared to hydrocodone by the intravenous route of administra hydrocodone penta-peptide conjugate, measured as free tion measured as free hydrocodone. hydrocodone. 25 FIG.55. Decrease in bioavailability ofYYFFI-HC as com FIG. 32. Intranasal bioavailability of an abuse-resistant pared to hydrocodone by the intravenous route of administra hydrocodone penta-peptide conjugate, measured as free tion measured as free hydrocodone. hydrocodone. FIG. 56. Oral bioavailability of hydrocodone plus hydro FIG. 33. Intranasal bioavailability of abuse-resistant morphone (concentration vs. time) following administration hydrocodone penta-peptide conjugates, measured as free 30 of hydrocodone bitratrate orYYFFI-HC at 1 mg/kg (equimo hydrocodone. lar doses with equivalent content of hydrocodone base) in FIG. 34. Intranasal bioavailability of an abuse-resistant rats, measured as free hydrocodone. hydrocodone penta-peptide conjugate, measured as free FIG. 57. Oral bioavailability of hydrocodone (concentra hydrocodone. tion vs. time) following administration of hydrocodone FIG. 35. illustrates preparation of 1.2:3,4-di-O-isopropy 35 bitratrate or YYFFI-HC at 1 mg/kg (equimolar doses with lidene-D-galactopyranose. equivalent content of hydrocodone base) in rats, measured as FIG. 36. Oral bioavailability of abuse-resistant hydroc free hydrocodone. odone glyco-peptide conjugates, measured as free hydroc FIG. 58. Oral bioavailability of hydromorphone (concen odone. tration vs. time) following administration of hydrocodone FIG. 37. Oral bioavailability of an abuse-resistant hydroc 40 bitratrate or YYFFI-HC at 1 mg/kg (equimolar doses with odone amino acid-carbohydrate conjugate, measured as free equivalent content of hydrocodone base) in rats, measured as hydrocodone. free hydrocodone. FIG. 38. illustrates nucleosides and conjugation sites. FIG. 59. Oral bioavailability of hydrocodone plus hydro FIG. 39. Oral bioavailability in rats for hydrocodone vs. morphone (concentration vs. time) following administration EEFFFI-HC at a dose (1 mg/kg) approximating a therapeutic 45 of hydrocodone bitratrate orYYFFI-HC at 2 mg/kg (equimo human dose equivalent measured as free hydrocodone. lar doses with equivalent content of hydrocodone base) in FIG. 40. Oral bioavailability in rats for hydrocodone vs. rats, measured as free hydrocodone. EEFFF-HC at a dose (1 mg/kg) approximating a therapeutic FIG. 60. Oral bioavailability of hydrocodone (concentra human dose equivalent measured as free hydrocodone. tion vs. time) following administration of hydrocodone FIG. 41. Oral bioavailability in rats for hydrocodone vs. 50 bitratrate or YYFFI-HC at 2 mg/kg (equimolar doses with YYI-HC at a dose (1 mg/kg) approximating a therapeutic equivalent content of hydrocodone base) in rats, measured as human dose equivalent measured as free hydrocodone. free hydrocodone. FIG. 42. Oral bioavailability in rats for hydrocodone vs. FIG. 61. Oral bioavailability of hydromorphone (concen DDI-HC at a dose (1 mg/kg) approximating a therapeutic tration vs. time) following administration of hydrocodone human dose equivalent measured as free hydrocodone. 55 bitratrate or YYFFI-HC at 2 mg/kg (equimolar doses with FIG. 43. Oral bioavailability in rats for hydrocodone vs. equivalent content of hydrocodone base) in rats, measured as YYFFI-HC at a dose (1 mg/kg) approximating a therapeutic free hydrocodone. human dose equivalent measured as free hydrocodone. FIG. 62. Oral bioavailability of hydrocodone plus hydro FIG. 44. Oral bioavailability in rats for hydrocodone vs. morphone (concentration vs. time) following administration EEFFI-HC at a dose (5 mg/kg) approaching a human over 60 of hydrocodone bitratrate orYYFFI-HC at 5 mg/kg (equimo dose equivalent measured as free hydrocodone. lar doses with equivalent content of hydrocodone base) in FIG. 45. Oral bioavailability in rats for hydrocodone vs. rats, measured as free hydrocodone. YYI-HC at a dose (5 mg/kg) approaching a human overdose FIG. 63. Oral bioavailability of hydrocodone (concentra equivalent measured as free hydrocodone. tion vs. time) following administration of hydrocodone FIG. 46. Oral bioavailability in rats for hydrocodone vs. 65 bitratrate or YYFFI-HC at 5 mg/kg (equimolar doses with DDI-HC at a dose (5 mg/kg) approaching a human overdose equivalent content of hydrocodone base) in rats, measured as equivalent measured as free hydrocodone. free hydrocodone. US 8,343,927 B2 5 6 FIG. 64. Oral bioavailability of hydromorphone (concen bitratrate or YYFFI-HC at 1 mg/kg (equimolar doses with tration vs. time) following administration of hydrocodone equivalent content of hydrocodone base) in rats, measured as bitratrate or YYFFI-HC at 5 mg/kg (equimolar doses with free hydrocodone. equivalent content of hydrocodone base) in rats, measured as FIG. 77. Intranasal bioavailability of hydromorphone free hydrocodone. (concentration vs. time) following administration of hydroc FIG. 65. Oral bioavailability of hydrocodone plus hydro odone bitratrate or YYFFI-HC at 1 mg/kg (equimolar doses morphone (concentration vs. time) following administration with equivalent content of hydrocodone base) in rats, mea of hydrocodone bitratrate or YYFFI-HC at 25 mg/kg Sured as free hydrocodone. (equimolar doses with equivalent content of hydrocodone FIG. 78. Oral bioavailability of hydrocodone plus hydro base) in rats, measured as free hydrocodone. 10 morphone (concentration vs. time) following administration FIG. 66. Oral bioavailability of hydrocodone (concentra of hydrocodone bitratrate orYYFFI-HC at 1 mg/kg (equimo tion vs. time) following administration of hydrocodone lar doses with equivalent content of hydrocodone base) in bitratrate or YYFFI-HC at 25 mg/kg (equimolar doses with rats, measured as free hydrocodone. equivalent content of hydrocodone base) in rats, measured as 15 FIG. 79. Oral bioavailability of hydrocodone (concentra free hydrocodone. tion vs. time) following administration of hydrocodone FIG. 67. Oral bioavailability of hydromorphone (concen bitratrate or YYFFI-HC at 1 mg/kg (equimolar doses with tration vs. time) following administration of hydrocodone equivalent content of hydrocodone base) in rats, measured as bitratrate or YYFFI-HC at 25 mg/kg (equimolar doses with free hydrocodone. equivalent content of hydrocodone base) in rats, measured as FIG. 80. Oral bioavailability of hydromorphone (concen free hydrocodone. tration vs. time) following administration of hydrocodone FIG. 68. Oral bioavailability (AUC) of hydrocodone bitratrate or YYFFI-HC at 1 mg/kg (equimolar doses with plus hydromorphone (concentration vs. dose) in proportion to equivalent content of hydrocodone base) in rats, measured as dose following administration of hydrocodone bitratrate or free hydrocodone. YYFFI-HC at escalating doses (1, 2, 5, and 25 mg/kg 25 FIG. 81. Oral bioavailability of hydrocodone plus hydro equimolar doses with equivalent content of hydrocodone morphone (concentration vs. time) following administration base) in rats, measured as free hydrocodone. of hydrocodone bitratrate orYYFFI-HC at 2 mg/kg (equimo FIG. 69. Oral bioavailability (AUC) of hydrocodone lar doses with equivalent content of hydrocodone base) in plus hydromorphone in proportion to human equivalent doses rats, measured as free hydrocodone. (HED) following administration of hydrocodone bitratrate or 30 FIG. 82. Oral bioavailability of hydrocodone (concentra YYFFI-HC at escalating doses (1, 2, 5, and 25 mg/kg equimolar doses with equivalent content of hydrocodone tion vs. time) following administration of hydrocodone base) in rats, measured as free hydrocodone. bitratrate or YYFFI-HC at 2 mg/kg (equimolar doses with FIG. 70. Oral bioavailability (C) of hydrocodone plus equivalent content of hydrocodone base) in rats, measured as hydromorphone (concentration VS. dose) in proportion to 35 free hydrocodone. dose following administration of hydrocodone bitratrate or FIG. 83. Oral bioavailability of hydromorphone (concen YYFFI-HC at escalating doses (1, 2, 5, and 25 mg/kg tration vs. time) following administration of hydrocodone equimolar doses with equivalent content of hydrocodone bitratrate or YYFFI-HC at 2 mg/kg (equimolar doses with base) in rats, measured as free hydrocodone. equivalent content of hydrocodone base) in rats, measured as FIG. 71. Oral bioavailability (C) of hydrocodone plus 40 free hydrocodone. hydromorphone in proportion to human equivalent doses FIG. 84. Oral bioavailability of hydrocodone plus hydro (HED) following administration of hydrocodone bitratrate or morphone (concentration vs. time) following administration YYFFI-HC at escalating doses (1, 2, 5, and 25 mg/kg of hydrocodone bitratrate orYYFFI-HC at 5 mg/kg (equimo equimolar doses with equivalent content of hydrocodone lar doses with equivalent content of hydrocodone base) in base) in rats, measured as free hydrocodone. 45 rats, measured as free hydrocodone. FIG. 72. Intravenous bioavailability of hydrocodone plus FIG. 85. Oral bioavailability of hydrocodone (concentra hydromorphone andYYFFI-HC (concentration vs. time) fol tion vs. time) following administration of hydrocodone lowing administration of hydrocodone bitratrate or YYFFI bitratrate or YYFFI-HC at 5 mg/kg (equimolar doses with HC at 1 mg/kg (equimolar doses with equivalent content of equivalent content of hydrocodone base) in rats, measured as hydrocodone base) in rats, measured as free hydrocodone. 50 free hydrocodone. FIG. 73. Intravenous bioavailability of hydrocodone (con FIG. 86. Oral bioavailability of hydromorphone (concen centration vs. time) following administration of hydrocodone tration vs. time) following administration of hydrocodone bitratrate or YYFFI-HC at 1 mg/kg (equimolar doses with bitratrate or YYFFI-HC at 5 mg/kg (equimolar doses with equivalent content of hydrocodone base) in rats, measured as equivalent content of hydrocodone base) in rats, measured as free hydrocodone. 55 free hydrocodone. FIG. 74. Intravenous bioavailability of hydromorphone FIG. 87. Oral bioavailability of hydrocodone plus hydro (concentration vs. time) following administration of hydroc morphone (concentration vs. time) following administration odone bitratrate or YYFFI-HC at 1 mg/kg (equimolar doses of hydrocodone bitratrate or YYFFI-HC at 25 mg/kg with equivalent content of hydrocodone base) in rats, mea (equimolar doses with equivalent content of hydrocodone Sured as free hydrocodone. 60 base) in rats, measured as free hydrocodone. FIG. 75. Intranasal bioavailability of hydrocodone plus FIG. 88. Oral bioavailability of hydrocodone (concentra hydromorphone (concentration vs. time) following adminis tion vs. time) following administration of hydrocodone tration of hydrocodone bitratrate or YYFFI-HC at 1 mg/kg bitratrate or YYFFI-HC at 25 mg/kg (equimolar doses with (equimolar doses with equivalent content of hydrocodone equivalent content of hydrocodone base) in rats, measured as base) in rats, measured as free hydrocodone. 65 free hydrocodone. FIG. 76. Intranasal bioavailability of hydrocodone (con FIG. 89. Oral bioavailability of hydromorphone (concen centration vs. time) following administration of hydrocodone tration vs. time) following administration of hydrocodone US 8,343,927 B2 7 8 bitratrate or YYFFI-HC at 25 mg/kg (equimolar doses with FIG. 106. Oral bioavailability of abuse-resistant oxyc equivalent content of hydrocodone base) in rats, measured as odone disubstituted tripeptide conjugates, measured as free free hydrocodone. oxycodone. FIG. 90. Oral bioavailability (AUC) of hydrocodone FIG. 107. Oral bioavailability of abuse-resistant oxyc plus hydromorphone (concentration vs. dose) in proportion to 5 odone disubstituted tripeptide conjugates, measured as free dose following administration of hydrocodone bitratrate or oxycodone. YYFFI-HC at escalating doses (1, 2, 5, and 25 mg/kg FIG. 108. Oral bioavailability of abuse-resistant oxyc equimolar doses with equivalent content of hydrocodone odone disubstituted tripeptide conjugates, measured as free base) in rats, measured as free hydrocodone. oxycodone. FIG. 91. Oral bioavailability (AUC) of hydrocodone 10 FIG. 109. Oral bioavailability of abuse-resistant oxyc plus hydromorphone in proportion to human equivalent doses odone disubstituted tripeptide conjugates, measured as free (HED) following administration of hydrocodone bitratrate or oxycodone. YYFFI-HC at escalating doses (1, 2, 5, and 25 mg/kg FIG. 110. Oral bioavailability of abuse-resistant oxyc equimolar doses with equivalent content of hydrocodone 15 odone disubstituted tripeptide conjugates, measured as free base) in rats, measured as free hydrocodone. oxycodone. FIG. 92. Oral bioavailability (C) of hydrocodone plus FIG. 111. Oral bioavailability of abuse-resistant oxyc hydromorphone (concentration VS. dose) in proportion to odone disubstituted tripeptide conjugates, measured as free dose following administration of hydrocodone bitratrate or oxycodone. YYFFI-HC at escalating doses (1, 2, 5, and 25 mg/kg. 20 FIG. 112. Oral bioavailability of abuse-resistant oxyc equimolar doses with equivalent content of hydrocodone odone disubstituted tripeptide conjugates, measured as free base) in rats, measured as free hydrocodone. oxycodone. FIG.93. Oral bioavailability (C) of hydrocodone plus FIG. 113. Oral bioavailability of abuse-resistant oxyc hydromorphone in proportion to human equivalent doses odone disubstituted tripeptide conjugates, measured as free (HED) following administration of hydrocodone bitratrate or 25 oxycodone. YYFFI-HC at escalating doses (1, 2, 5, and 25 mg/kg FIG. 114. Oral bioavailability of abuse-resistant oxyc equimolar doses with equivalent content of hydrocodone odone disubstituted tripeptide conjugates, measured as free base) in rats, measured as free hydrocodone. oxycodone. FIG. 94. Intravenous bioavailability of hydrocodone plus FIG. 115. Oral bioavailability of abuse-resistant oxyc hydromorphone andYYFFI-HC (concentration vs. time) fol- 30 odone disubstituted tripeptide conjugates, measured as free lowing administration of hydrocodone bitratrate or YYFFI oxycodone. HC at 1 mg/kg (equimolar doses with equivalent content of FIG. 116. Oral bioavailability of abuse-resistant oxyc hydrocodone base) in rats, measured as free hydrocodone. odone disubstituted tripeptide conjugates, measured as free FIG.95. Intravenous bioavailability of hydrocodone (con oxycodone. centration vs. time) following administration of hydrocodone 35 FIG. 117. Oral bioavailability of abuse-resistant oxyc bitratrate or YYFFI-HC at 1 mg/kg (equimolar doses with odone disubstituted tripeptide conjugates, measured as free equivalent content of hydrocodone base) in rats, measured as oxycodone. free hydrocodone. FIG. 118. Oral bioavailability of abuse-resistant oxyc FIG. 96. Intravenous bioavailability of hydromorphone odone disubstituted tripeptide conjugates, measured as free (concentration vs. time) following administration of hydroc- 40 oxycodone. odone bitratrate or YYFFI-HC at 1 mg/kg (equimolar doses FIG. 119. Oral bioavailability of abuse-resistant oxyc with equivalent content of hydrocodone base) in rats, mea odone disubstituted tripeptide conjugates, measured as free Sured as free hydrocodone. oxycodone. FIG. 97. Intranasal bioavailability of hydrocodone plus FIG. 120. Oral bioavailability of abuse-resistant oxyc hydromorphone (concentration vs. time) following adminis- 45 odone disubstituted tripeptide conjugates, measured as free tration of hydrocodone bitratrate or YYFFI-HC at 1 mg/kg oxycodone. (equimolar doses with equivalent content of hydrocodone FIG. 121. Oral bioavailability of abuse-resistant oxyc base) in rats, measured as free hydrocodone. odone disubstituted tripeptide conjugates, measured as free FIG. 98. Intranasal bioavailability of hydrocodone (con oxycodone. centration vs. time) following administration of hydrocodone 50 FIG. 122. Oral bioavailability of abuse-resistant oxyc bitratrate or YYFFI-HC at 1 mg/kg (equimolar doses with odone disubstituted tripeptide conjugates, measured as free equivalent content of hydrocodone base) in rats, measured as oxycodone. free hydrocodone. FIG. 123. Oral bioavailability of abuse-resistant oxyc FIG. 99. Intranasal bioavailability of hydromorphone odone disubstituted tripeptide conjugates, measured as free (concentration vs. time) following administration of hydroc- 55 oxycodone. odone bitratrate or YYFFI-HC at 1 mg/kg (equimolar doses FIG. 124. Intranasal bioavailability of abuse-resistant oxy with equivalent content of hydrocodone base) in rats, mea codone disubstituted tripeptide conjugates, measured as free Sured as free hydrocodone. oxycodone. FIG. 100. depicts oxycodone. FIG. 125. Intranasal bioavailability of abuse-resistant oxy FIG. 101. depicts oxycodone with lysine branched pep- 60 codone disubstituted tripeptide conjugates, measured as free tides. oxycodone. FIG. 102. depicts a glycosylated oxycodone. FIG. 126. Intranasal bioavailability of abuse-resistant oxy FIG. 103. depicts formation of an enol ether with serine. codone disubstituted tripeptide conjugates, measured as free FIG. 104. depicts niacin and biotin. oxycodone. FIG. 105. Oral bioavailability of abuse-resistant oxyc- 65 FIG. 127. Intravenous bioavailability of abuse-resistant odone disubstituted tripeptide conjugates, measured as free oxycodone disubstituted tripeptide conjugates, measured as oxycodone. free oxycodone. US 8,343,927 B2 10 FIG. 128. Intranasal bioavailability of abuse-resistant oxy agent conjugate is Substantially decreased as compared to the codone disubstituted tripeptide conjugates, measured as free parent active agent. The relative decrease in bioavailability at oxycodone. higher doses abates the euphoria obtained when doses of the FIG. 129. Intranasal bioavailability of abuse-resistant oxy active agent conjugate are taken above those of the intended codone disubstituted tripeptide conjugates, measured as free 5 prescription. This in turn diminishes the abuse potential, oxycodone. whether unintended or intentionally sought. FIG. 130. Intranasal bioavailability of abuse-resistant oxy Persons that abuse prescription drugs commonly seek to codone disubstituted tripeptide conjugates, measured as free increase their euphoria by Snorting or injecting the drugs. oxycodone. These routes of administration increase the rate and extent of FIG. 131. Intranasal bioavailability of abuse-resistant oxy 10 drug absorption and provide a faster, nearly instantaneous, codone disubstituted tripeptide conjugates, measured as free oxycodone. effect. This increases the amount of drug that reaches the FIG. 132. Intranasal bioavailability of abuse-resistant oxy central nervous system where it has its effect. In a particular codone disubstituted tripeptide conjugates, measured as free embodiment of the invention the bioavailability of the oxycodone. 15 covalently modified active agent is Substantially decreased by FIG. 133. Intranasal bioavailability of abuse-resistant oxy the intranasal and intravenous routes as compared to the par codone disubstituted tripeptide conjugates, measured as free ent active agent. Thus the illicit practice of Snorting and oxycodone. shooting the drug loses its advantage. FIG. 134. Intranasal bioavailability of abuse-resistant oxy In accordance with the present invention and as used codone disubstituted tripeptide conjugates, measured as free herein, the following terms are defined with the following oxycodone. meanings, unless explicitly stated otherwise. For additional FIG. 135. Intranasal bioavailability of abuse-resistant oxy methods of attaching active agents to carriers, see U.S. appli codone disubstituted tripeptide conjugates, measured as free cation Ser. No. 10/156,527, and/or PCT/US03/05524, and/or oxycodone. PCT/USO3/05525 and/or PCT/USO4f17204 each of which is FIG. 136. Intranasal bioavailability of abuse-resistant oxy 25 hereby incorporated by reference in its entirety. In the present codone disubstituted tripeptide conjugates, measured as free invention, dextroamphetamine is covalently attached to the oxycodone. peptide via the amino group. FIG. 137. Intranasal bioavailability of abuse-resistant oxy The invention utilizes covalent modification of an active codone disubstituted tripeptide conjugates, measured as free agent to decrease its potential for causing overdose or being oxycodone. 30 abused. The active agent is covalently modified in a manner FIG. 138. Intranasal bioavailability of abuse-resistant oxy that decreases its pharmacological activity, as compared to codone disubstituted tripeptide conjugates, measured as free the unmodified active agent, at doses above those considered oxycodone. therapeutic, e.g., at doses inconsistent with the manufactur FIG. 139. Intranasal bioavailability of abuse-resistant oxy er's instructions. When given at lower doses. Such as those codone disubstituted tripeptide conjugates, measured as free 35 intended for therapy, the covalently modified active agent oxycodone. retains pharmacological activity similar to that of the unmodi FIG. 140. Intranasal bioavailability of abuse-resistant oxy fied active agent. The covalent modification of the active codone disubstituted tripeptide conjugates, measured as free agent may comprise the attachment of any chemical moiety oxycodone. through conventional chemistry. FIG. 141. Intranasal bioavailability of abuse-resistant oxy 40 Compounds, compositions and methods of the invention codone disubstituted tripeptide conjugates, measured as free provide reduced potential for overdose, reduced potential for oxycodone. abuse or addiction and/or improve the active agent's charac FIG. 142. Oral bioavailability in rats of oxycodone vs. teristics with regard to high toxicities or suboptimal release P2L2-Oxycodone at a dose (2.5 mg/kg) approximating a profiles. therapeutic human dose equivalent measured as free oxyc 45 The following is a non-exclusive list of compounds that odone. may be used in connection with the invention: alphacetyl FIG. 143. Decrease in bioavailability of P2L-Oxyc methadol hydrochloride, anilleridine, apomorphine, bemi odone as compared to oxycodone by the intranasal route of done, betacetylmethadol hydrochloride, buprenorphine administration-dose 2.5 mg/kg measured as free oxycodone. hydrochloride, butorphanol tartrate, codeine, dezocine, dihy FIG. 144. Decrease in bioavailability of P2L-Oxyc 50 drocodeine, dihydromorphine, dipanone hydrochloride, epta odone as compared to oxycodone by the intravenous route of Zocine hydrobromide, ethylmorphine, etorphine hydrochlo administration-dose 0.5 mg/kg measured as free oxycodone. ride, hydromorphone, ketobemidone, levorphanol tartrate, loperamide, meptazinol hydrochloride, methyldihydromor DETAILED DESCRIPTION OF THE INVENTION phinone, nalbuphine hydrochloride, nalbuphine hydrochlo 55 ride, normorphine, oxycodone, Oxymorphone, pentazocine, The present invention relates to changing the pharmacoki piminodine, tramadol, allobarbitone, alprazolan, amylobar netic and pharmacological properties of active agents through bitone, barbitone sodium, butobarbitone, captodiame hydro covalent modification. Covalent attachment of a chemical chloride, chloral betaine, chloral hydrate, chloralose, chlo moiety to an active agent can change the rate and extent of rhexadol, chlormethiazole edisylate, cinolazepam, potassium absorption, metabolism, distribution, and elimination of the 60 cloraZepate, cyclobarbitone calcium, delorZepam, difebar active agent. When administered at a normal therapeutic dose bamate, enciprazine hydrochloride, flunitrazepam, hexobar the bioavailability (area under the time-versus-concentration bitone sodium, ibomal, lorazepam, lormetazepam, mep curve; AUC) of the active agent is similar to that of the parent robamate, methylpentynol, midazolam maleate, oxazepam, active agent compound. As the oral dose is increased, how pentabarbitone calcium, phenprobamate, proxibarbal, ever, the bioavailability of the covalently modified active 65 quinalbaritone, quinalbarbitone sodium, secbutobarbitone agent relative to the parent active agent begins to decline. At Sodium, temazepam, triclofos sodium, Zalepan, and Suprapharmacological doses the bioavailability of the active Zolazepam hydrochloride. US 8,343,927 B2 11 12 Throughout this application the use of “opioid' is meant to glycosyl-amino-acid is a compound consisting of saccharide include any drug that activates the opioid receptors found in linked through a glycosyl linkage (O—, N— or S-) to an the brain, spinal cord and gut. There are three broad classes of amino acid. opioids: naturally occurring opium alkaloids, such as mor A “composition” as used herein, refers broadly to any phine (the prototypical opioid) and codeine; semi-synthetics composition containing a described molecule conjugates. Such as heroine, oxycodone and hydrocodone that are pro The composition may comprise a dry formulation, an aque duced by modifying natural opium alkaloids and have similar ous Solution, or a sterile composition. Compositions compris chemical structures; and pure synthetics Such as fentanyl and ing the molecules described herein may be stored in freeze methadone that are not produced from opium and may have dried form and may be associated with a stabilizing agent very different chemical structures than the opium alkaloids. 10 Such as a carbohydrate. In use, the composition may be Other opioids include hydroxymorphone, oxymorphone, deployed in an aqueous solution containing salts, e.g., NaCl, methadone, levorphanol, dihydrocodeine, meperidine, diphe detergents, e.g., Sodium dodecyl sulfate (SDS), and other noxylate, Sufentanil, alfentanil, propoxyphene, pentazocine, components. nalbuphine, butorphanol, buprenorphine, meptazinol, dezo A “controlled substance' is a substance subject to federal cine, and pharmaceutically acceptable salts thereof. 15 regulation of its manufacture, sale, or distribution because of Throughout this application the use of “oxycodone” is the potential for, or proved evidence of abuse; because of its meant to include a narcotic alkaloid (chemical formula potential for psychic or physiological dependence; because it CHNO) and its derivatives such as the hydrochloride salt constitutes a public health risk; because of the scientific evi of oxycodone. Oxycodone is related to codeine and is used as dence of its pharmacologic effect; or because of its role as a an analgesic and/or a sedative. Oxycodone is a powerful and precursor of other controlled substances. potentially addictive opioid analgesic synthesized from the Important note regarding stereochemistry: This patent is baine. It is similar to codeine, but is more potent and has a meant to cover all compounds discussed regardless of abso higher dependence potential. It is effective orally and is often lute configurations. Thus, natural, L-amino acids are dis marketed in combination with aspirin (Percodan R) or cussed but the use of D-amino acids are also included. acetaminophen (Percocet(R) for the relief of pain. It is also 25 The following abbreviations may be in this application: sold in a Sustained-release form under the trade name Oxy BOC-t-butyloxycarbonyl contin R. All of these deriviatives or combinations of oxyc CMC-carboxymethylcellulose odone are encompassed by the present invention. DIPEA-di-isopropyl ethylamine Throughout this application the use of “hydrocodone” is mp-melting point meant to include a semisynthetic narcotic analgesic and anti 30 NMR-nuclear magnetic resonance tussive prepared from codeine with multiple actions qualita OSu-hydroxysuccinimido ester tively similar to those of codeine. It is commonly used for the Nia=Niacin relief of moderate to moderately severe pain. Trade names Bio=Biotin include AnexsiaR), Hycodan R), Hycomine(R), Lorcet(R), The attached chemical moiety may be any chemical Sub Lortab(R), Norco(R, Tussionex R, Tylox R, and Vicodin(R). 35 stance that decreases the pharmacological activity until the Derivatives of hydrocodone, such as hydrocodone bitartrate active agent is released. Preferably the chemical moiety is a and hydrocodone polistirex, are encompassed by the present single amino acid, dipeptide or tripeptide, tetrapeptide, pen invention. tapeptide, or hexapeptide. The active agent binds to specific Throughout this application the use of "peptide' is meant sites to produce various effects (Hoebel, et al., 1989). The to include a single amino acid, a dipeptide, a tripeptide, an 40 attachment of certain chemical moieties can therefore dimin oligopeptide, a polypeptide, or the carrier peptide. Oligopep ish or prevent binding to these biological target sites. Prefer tide is meant to include from 2 amino acids to 70amino acids. ably, absorption of the composition into the brain is prevented Further, at times the invention is described as being an active or substantially diminished and/or delayed when delivered by agent attached to an amino acid, a dipeptide, a tripeptide, an routes other than oral administration. oligopeptide, or polypeptide to illustrate specific embodi 45 The attached chemical moiety may further comprise natu ments for the active agent conjugate. Preferred lengths of the rally occurring or synthetic Substances. This would include conjugates and other preferred embodiments are described but is not limited to the attachment of an active agent to one or herein. more amino acids, peptides, lipids, carbohydrates, glycopep Throughout this application the use of “chemical moiety’ tides, nucleic acids or vitamins. These chemical moieties is meant to include at least amino acids, peptides, glycopep 50 could be expected to affect delayed release in the gastrointes tides, carbohydrates, lipids, nucleosides, or vitamins. tinal tract and prevent rapid onset of the desired activity, “Carbohydrates' includes Sugars, starches, cellulose, and particularly when delivered by parenteral routes. (Hoebel, B. related compounds. e.g., (CHO), wherein n is an integer G. L. Hernandez, et al. (1989). “Microdialysis studies of larger than 2 or C(HO), with n larger than 5. More spe brain norepinephrine, serotonin, and dopamine release during cific examples include for instance, fructose, glucose, lactose, 55 ingestive behavior. Theoretical and clinical implications.” maltose, Sucrose, glyceraldehyde, dihydroxyacetone, eryth Ann NY AcadSci 575: 171-91). rose, ribose, ribulose, Xylulose, galactose, mannose, Sedohep For each of the embodiments recited herein, the amino acid tulose, neuraminic acid, dextrin, and glycogen. or peptide may comprise of one or more of the naturally A glycoprotein’ is a compound containing carbohydrate occurring (L-) amino acids: alanine, arginine, asparagine, (or glycan) covalently linked to protein. The carbohydrate 60 aspartic acid, cysteine, glycine, glutamic acid, glutamine, may be in the form of a monosaccharide, disaccharide(s). histidine, isoleucine, leucine, lysine, methionine, proline, oligosaccharide(s), polysaccharide(s), or their derivatives phenylalanine, serine, tryptophan, threonine, tyrosine, and (e.g. Sulfo- or phospho-substituted). Valine. In another embodiment the amino acid or peptide is A glycopeptide' is a compound consisting of carbohy comprised of one or more of the naturally occurring (D) drate linked to an oligopeptide composed of L- and/or 65 amino acids: alanine, arginine, asparagine, aspartic acid, cys D-amino acids. A glyco-amino-acid is a saccharide attached teine, glycine, glutamic acid, glutamine, histidine, isoleucine, to a single amino acid by any kind of covalent bond. A leucine, lysine, methionine, proline, phenylalanine, serine, US 8,343,927 B2 13 14 tryptophan, threonine, tyrosine, and valine. In another Exemplary nonsteroidal anti-inflammatory agents include embodiment the amino acid or peptide is comprised of one or ibuprofen, naproxen or indomethacin, aspirin or a salicylic more unnatural, non-standard or synthetic amino acids Such acid derivative, or acetaminophen. as, aminohexanoic acid, biphenylalanine, cyclohexylalanine, Exemplary anti-depressants include citalopram, fluoxet cyclohexylglycine, diethylglycine, dipropylglycine, 2,3-di ine, norfluoxetine, fluvoxamine, paroxetine, Sertraline, ami aminoproprionic acid, homophenylalanine, homoserine, triptyline, desipramine, doxepin, imipramine, nortryiptyline, homotyrosine, naphthylalanine, norleucine, ornithine, phey bupropion, mirtazapine, nefazodone, traZodone, or Venlafax lalanine(4-fluoro), phenylalanine(2,3,4,5,6 pentafluoro), 1C. phenylalanine(4-nitro), phenylglycine, pipecolic acid, sar Exemplary anti-psychotics include clozapine, haloperidol, cosine, tetrahydroisoquinoline-3-carboxylic acid, and tert 10 olanzapine, quetiapine, or risperidone. leucine. In another embodiment the amino acid or peptide comprises of one or more amino acid alcohols. In another The compositions and methods of the invention provide embodiment the amino acid or peptide comprises of one or active agents which when bound to the chemical moiety pro more N-methyl amino acids. vide safer and/or more effective dosages for the above recited In another embodiment, the specific carriers are utilized as 15 active agent classes through improved bioavailability curves a base short chain amino acid sequence and additional amino and/or safer C, and/or reduce area under the curve for acids are added to the terminus or side chain. In another bioavailability, particularly for abused substances taken in embodiment, the above amino acid sequence may have one doses above therapeutic levels. As a result, the compositions more of the amino acids substituted with one of the 20 natu and methods of the invention may provide improved methods rally occurring amino acids. It is preferred that the Substitu of treatment for attention deficit hyperactivity, attention defi tion be with an amino acid which is similar in structure or cit hyperactivity disorder (ADHD), attention deficit disorder charge compared to the amino acid in the sequence. For (ADD), cognitive decline associated with acquired immuno instance, isoleucine (Ile)I is structurally very similar to leu deficiency syndrome (AIDS) or AIDS-related complex, cine (Leu) L, whereas, tyrosine (Tyr)Y is similar to phe depression, anxiety and anxiety related disorders, psychosis, nylalanine (Phe) F, whereas serine (Ser)SI is similar to 25 nicotine addiction, narcotic addiction, alcoholism, narco threonine (Thr)T, whereas cysteine (Cys)C is similar to lepsy, and/or analgesia. methionine (Met)M), whereas alanine (Ala)A is similar to In one embodiment the chemical moiety is comprised of an valine (Val)V, whereas lysine (Lys) K is similar to arginine amino acid or a polypeptide. Preferred amino acid and pep (Arg) R, whereas asparagine (Asn)N is similar to tide chemical moieties include, for example, Lys, Ser, Ala, glutamine (Gln)Q, whereas aspartic acid (Asp)D is simi 30 Phe, Ile, Pro-Pro-Leu, Pro-Pro-Ile, Val-Val. Lys-Lys, Gly lar to glutamic acid (Glu)|E|, whereas histidine (His)H is Gly-Ile, Phe-Phe-Leu, Thr-Thr-Val, Tyr-Tyr-Val, Tyr-Tyr similar to proline (Pro)P, and glycine (Gly) G is similar to Phe, Glu-Glu-Val. Asp-Asp-Val. Lys-Lys-Val. Glu-Glu-Phe tryptophan (Trp)W. In the alternative the preferred amino Phe-Ile, Glu-Glu-Phe-Phe-Phe, Tyr-Tyr-Ile, Asp-Asp-Ile, acid Substitutions may be selected according to hydrophilic Tyr-Tyr-Phe-Phe-Ile, Tyr-Tyr-Lys-Tyr-Tyr, Phe-Phe-Lys properties (i.e. polarity) or other common characteristics 35 associated with the 20 essential amino acids. While preferred Phe-Phe, Glu-Glu-Phe-Phe-Ile, (Lys-Lys-Gly-Gly), and embodiments utilize the 20 natural amino acids for their (1)-Lys-(d)-Lys-Leu. In some embodiments, the active GRAS characteristics, it is recognized that minor substitu agent is disubstituted with one or more of the preceding tions along the amino acid chain which do not effect the chemical moieties. essential characteristics of the amino are also contemplated. 40 Another embodiment of the invention is a composition for In one embodiment the carrier range is between one to 12 preventing overdose comprising an active agent which has chemical moieties with one to 8 moieties being preferred. In been covalently bound to a chemical moiety. another embodiment the number of chemical moieties Another embodiment of the invention is a composition for attached is selected from 1, 2, 3, 4, 5, 6, or 7, etc. In another safely delivering an active agent comprising providing a embodiment of the invention the molecular weight of the 45 therapeutically effective amount of said active agent which carrier portion of the conjugate is below about 2,500, more has been covalently bound to a chemical moiety wherein said preferably below about 1,000 and most preferably below chemical moiety reduces the rate of absorption of the active about 500. agent as compared to delivering the unbound active agent. The compositions and methods of the invention may be Another embodiment of the invention is a composition for applied to various therapeutically valuable active agents (e.g., 50 reducing drug toxicity comprising providing a patient with an drugs) and include, for example, stimulants such as amphet active agent which has been covalently bound to a chemical amines, anticonvulsants, muscle relaxants, antidepressants, moiety wherein said chemical moiety increases the rate of anxiolytics, benzodiazepines, sedatives, hypnotics, narcotics, clearance of an active agent when given at doses exceeding steroids, respiratory agents, including antihistamines, antip those within the therapeutic range of said active agent. sychotics including risperidone, and nonsteroidal anti-in 55 Another embodiment of the invention is a composition for flammatory agents. reducing drug toxicity comprising providing a patient with an Exemplary narcotics include opioids, hydrocodone, oxyc active agent which has been covalently bound to a chemical odone, morphine, codeine, hydroxymorphone, oxymor moiety wherein said chemical moiety provides a serum phone, methadone, fentanyl, levorphanol, dihydrocodeine, release curve which does not increase above said active agent meperidine, diphenoxylate, Sufentanil, alfentanil, pro 60 toxicity level when given at doses exceeding those within the poxyphene, pentazocine, nalbuphine, butorphanol, buprenor therapeutic range of said active agent. phine, meptazinol, dezocine or pharmaceutically acceptable Another embodiment of the invention is a composition for salts thereof. reducing bioavailability of active agent comprising active Exemplary benzodiazepines include alprazolam, chlor agent covalently bound to a chemical moiety wherein said diazepoxide, clonazepam, cloraZepate, diazepam, estazolam, 65 bound active agent maintains a steady-state serum release flurazepam, halazepam, lorazepam, midazolam, oxazepam, curve which provides a therapeutically effective bioavailabil quaZepam, temazepam, or triazolam. ity but prevents spiking or increase blood serum concentra US 8,343,927 B2 15 16 tions compared to unbound active agent when given at doses Arg, Phe-Cys, Phe-Gln, Phe-Tyr, Val-Glu, Val-Ser, Val-Asp, exceeding those within the therapeutic range of said active Val-Asn. Val-Thr, Val-Arg, Val-Cys, Val-Gln, or Val-Tyr. agent. In one embodiment, the —X—Z component of the formula Another embodiment of the invention is a composition for A-X, Z comprises Pro-Pro-Leu, Pro-Pro-He, Val-Val. preventing a C, spike for active agent while still providing Lys-Lys, Gly-Gly-Ile, Phe-Phe-Ile, Phe-Phe-Leu, Thr-Thr a therapeutically effective bioavailability curve comprising Val, Tyr-Tyr-Val, Tyr-Tyr-Phe, Glu-Glu-Val. Asp-Asp-Val. an active agent which has been covalently bound to a chemi Lys-Lys-Val, Glu-Glu-Phe-Phe-Ile, Glu-Glu-Phe-Phe-Phe, cal moiety. Tyr-Tyr-Ile, Asp-Asp-Ile, Tyr-Tyr-Phe-Phe-Ile, Tyr-Tyr-Lys Another embodiment of the invention is a composition for Tyr-Tyr, Phe-Phe-Lys-Phe-Phe, Glu-Glu-Phe-Phe-Ile, (Lys preventing a toxic release profile in a patient comprising 10 Lys-Gly-Gly), or (1)-Lys-(d)-Lys-Leu. In one embodiment, the —X—Z component of the formula active agent covalently bound to a chemical moiety wherein A-X, Z comprises Ser-Ser, PolySer, Glu-Glu, Asp-Asp, said bound active agent maintains a steady-state serum Asp-Asp-Asp, Asp-Asp-Glu, Asp-Asp-Ser, Asp-Asp-LyS, release curve which provides a therapeutically effective bio Asp-Asp-Cys, Ala-Glu, Ala-Ser, Ala-Asp, Ala-ASn, Ala-Thr, availability but prevents spiking or increase blood serum con 15 Ala-Arg, Ala-Cys, Ala-Gln, Ala-Tyr, Leu-Glu, Leu-Ser, Leu centrations compared to unbound active agent. Asp, Leu-ASn, Leu-Thr, Leu-Arg, Leu-Cys, Leu-Gln, Leu Another embodiment of the invention is a compound of Tyr, Phe-Glu, Phe-Ser, Phe-Asp, Phe-ASn, Phe-Thr, Phe-Arg, Formula I: Phe-Cys, Phe-Gln, Phe-Tyr, Val-Glu, Val-Ser, Val-Asp, Val ASn, Val-Thr, Val-Arg, Val-Cys, Val-Gln, Val-Tyr, Ala-Pro, A-X. Z. Gly-Gly-Leu, Gly-Gly-Gly-Gly-Leu, Glu-Glu-Phe-Phe wherein A is active agent as defined herein; X is a chemical Phe-Ile, Glu-Glu-Phe-Phe-Phe, Tyr-Tyr-Ile, Asp-Asp-Ile, moiety as defined herein and n is between 1 and 50 and Tyr-Tyr-Phe-Phe-Ile, or Glu-Glu-Phe-Phe-Ile. increments therein; and Z is a further chemical moiety differ In one embodiment, the —X—Z component of the formula ent from X which acts as an adjuvant and m is between 1 and A-X, Z comprises Ser-Ser, PolySer, Lys-Lys, Lys-Lys 50 and increments therein. In another embodiment n is 25 Val, (Lys-Lys-Gly-Gly), (1)-Lys-(d)-Lys-Leu, Glu-Glu, between 1 and 10 and m is 0. It should be recognized that the Glu-Glu-Phe-Phe-Phe-Ile, Glu-Glu-Phe-Phe-Phe, Glu-Glu compounds of this formula may be used alone or in combi Phe-Phe-Ile, Glu-Glu-Val. Asp-Asp, Asp-Asp-Asp, Asp nation with any of the recited embodiments of the invention. Asp-Glu, Asp-Asp-Ser, Asp-Asp-LyS, Asp-Asp-Cys, Asp In one embodiment, a compound of the formula: A-X - Asp-Val, Ala-Glu, Ala-Pro, Ala-Ser, Ala-Asp, Ala-Asn, Ala Z, wherein A is an active agent; X is a linker covalently bound 30 Thr, Ala-Arg, Ala-Cys, Ala-Gln, Ala-Tyr, Leu-Glu, Leu-Ser, to A; and Z is an amino acid, peptide, or oligopeptide Leu-Asp, Leu-ASn, Leu-Thr, Leu-Arg, Leu-Cys, Leu-Gln, covalently bound to X, wherein mand n range from 1 to 50 is Leu-Tyr, Phe-Glu, Phe-Ser, Phe-Asp, Phe-ASn, Phe-Thr, utilized in connection with the present invention. It should be Phe-Arg, Phe-Cys, Phe-Gln, Phe-Tyr, Phe-Phe-Ile, Phe-Phe recognized that the formula A-X, Z generally describes Leu, Phe-Phe-Lys-Phe-Phe, Val-Glu, Val-Ser, Val-Asp, Val the components making up the compound and does not imply 35 ASn, Val-Thr, Val-Arg, Val-Cys, Val-Gln, Val-Tyr, Val-Val. any particular order in the attachment or type of bond between Gly-Gly-Leu, Gly-Gly-Gly-Gly-Leu, Gly-Gly-Ile, Tyr-Tyr the elements A, X, or Z. In the above formula X may comprise Ile, Tyr-Tyr-Phe-Phe-Ile, Tyr-Tyr-Val, Tyr-Tyr-Phe, Tyr-Tyr a small linear or cyclic molecule containing 2-6 atoms with Lys-Tyr-Tyr, Asp-Asp-Ile, Pro-Pro-Leu, Pro-Pro-Ile, or Thr one or more heteroatoms and one or more functional groups. Thr-Val. The functional groups may be selected from amines, amides, 40 In one embodiment, the component A of the formula alcohols or acids. A may be oxycodone or hydrocodone. X A-X, Z is a pain relief drug such as Oxycodone or hydro may be an enolate oran amino acid. In one embodiment, Z is codone. The component A of the formula A-X, Z may also bonded to X through a side chain of the amino acid. In another be alphacetylmethadol hydrochloride, anileridine, apomor embodiment, Z may be bonded to X through a peptide bond. phine, bemidone, betacetylmethadol hydrochloride, The linker X may comprise the following amino acids: Ser, 45 buprenorphine hydrochloride, butorphanol tartrate, codeine, Lys, Glu, Asp, Ala, Leu, Phe, Val, Gly, Tyr, Pro, or Thr. In one dezocine, dihydrocodeine, dihydromorphine, dipanone embodiment, where X is lysine, Z is bonded to lysine through hydrochloride, eptazocine hydrobromide, ethylmorphine, the side chain of the lysine. In another embodiment, where X etorphine hydrochloride, hydromorphone, ketobemidone, is lysine, Z is bonded to lysine through a peptide bond. levorphanol tartrate, loperamide, meptazinol hydrochloride, In one embodiment, the —X—Z component of the formula 50 methyldihydromorphinone, nalbuphine hydrochloride, nal A-X, Z comprises Lys-Lys, Lys-Lys-Val. (Lys-Lys-Gly buphine hydrochloride, normorphine, oxycodone, oxymor Gly), (1)-Lys-(d)-Lys-Leu, Glu-Glu, Glu-Glu-Phe-Phe phone, pentazocine, piminodine, tramadol, allobarbitone, Phe-Ile, Glu-Glu-Phe-Phe-Phe, Glu-Glu-Phe-Phe-Ile, Glu alprazolan, amylobarbitone, barbitone sodium, butobarbi Glu-Val. Asp-Asp, Asp-Asp-Asp, Asp-Asp-Glu, Asp-Asp tone, captodiame hydrochloride, chloral betaine, chloral Ser, Asp-Asp-LyS, Asp-Asp-Cys, Asp-Asp-Val. Ala-Ser, Ala 55 hydrate, chloralose, chlorhexadol, chlormethiazole edisylate, Cys, Leu-Glu, Leu-Ser, Leu-Asp, Leu-ASn, Leu-Thr, Leu cinolazepam, potassium cloraZepate, cyclobarbitone cal Arg, Leu-Cys, Leu-Gln, Leu-Tyr, Val-Glu, Val-Ser, Val-Asp, cium, delorZepam, difebarbamate, enciprazine hydrochlo Val-Asn. Val-Thr, Val-Arg, Val-Cys, Val-Gln, Val-Tyr, Val ride, flunitrazepam, hexobarbitone sodium, ibomal, Val, Gly-Gly-Leu, Gly-Gly-Gly-Gly-Leu, Gly-Gly-Ile, Asp lorazepam, lormetazepam, meprobamate, methylpentynol, Asp-Ile, Pro-Pro-Leu, Pro-Pro-Ile, or Thr-Thr-Val. 60 midazolam maleate, oxazepam, pentabarbitone calcium, In one embodiment, the —X—Z component of the formula phenprobamate, proxibarbal, quinalbaritone, quinalbarbi A-X, Z comprises Ser-Ser, PolySer, Lys, Glu-Glu, Asp tone sodium, secbutobarbitone sodium, temazepam, triclofoS Asp, Asp-Asp-Asp, Asp-Asp-Glu, Asp-Asp-Ser, Asp-Asp Sodium, Zalepan, or Zolazepam hydrochloride. LyS, Asp-Asp-Cys, Ala-Glu, Ala-Ser, Ala-Asp, Ala-ASn, Ala In one embodiment, the compound A-X, Z may be a Thr, Ala-Arg, Ala-Cys, Ala-Gln, Ala-Tyr, Leu6lu, Leu-Ser, 65 component of a pharmaceutical composition including the Leu-Asp, Leu-ASn, Leu-Thr, Leu-Arg, Leu-Cys, Leu-Gln, compound and a pharmaceutically acceptable excipient. The Leu-Tyr, PheGlu, Phe-Ser, Phe-Asp, Phe-ASn, Phe-Thr, Phe compound A-X, Z may be administered to a mammal in a US 8,343,927 B2 17 18 therapeutically effective amount to treat pain. The mammal Cys, Phe-Gln, Phe-Tyr, Val-Glu, Val-Ser, Val-Asp, Val-Asn. may be a human, primate, equine, canine, or feline. Val-Thr, Val-Arg, Val-Cys, Val-Gln, Val-Tyr, Ala-Pro, Gly In one embodiment, a compound of the formula: A-X - Gly-Leu, Gly-Gly-Gly-Gly-Leu, Glu-Glu-Phe-Phe-Phe-Ile, Z, wherein A is a drug radical; X is a small linear or cyclic Glu-Glu-Phe-Phe-Phe, Tyr-Tyr-Ile, Asp-Asp-Ile, Tyr-Tyr molecule containing 2-6 atoms with one or more heteroatoms Phe-Phe-Ile, and Glu-Glu-Phe-Phe-Ile, wherein an optional and one or more functional groups, wherein X is bonded to A linker attaches the first amino acid to the drug radical may be through a carboxyl group; and Z is an amino acid, peptide, or utilized in connection with the present invention. oligopeptide covalently bound to X, wherein m and n range In one embodiment, a compound comprising a drug radical between 1 and 50 is utilized in connection with the present covalently bonded to a peptide selected from the group con invention. 10 sisting of Ser-Ser, PolySer, Lys-Lys, Lys-Lys-Val. (Lys-Lys In one embodiment, a method of making a drug more Gly-Gly), (1)-Lys-(d)-Lys-Leu, Glu-Glu, Glu-Glu-Phe difficult to abuse, comprising bonding to a nucleophile on the Phe-Phe-Ile, Glu-Glu-Phe-Phe-Phe, Glu-Glu-Phe-Phe-Ile, drug a radical of the formula—X—Z, wherein X is a linker; Glu-Glu-Val. Asp-Asp, Asp-Asp-Asp, Asp-Asp-Glu, Asp and Z is an amino acid or peptide covalently bound to X may Asp-Ser, Asp-Asp-Lys, Asp-Asp-Cys, Asp-Asp-Val. Ala be utilized. 15 Glu, Ala-Pro, Ala-Ser, Ala-Asp, Ala-ASn, Ala-Thr, Ala-Arg, In one embodiment, a compound comprising a drug radical Ala-Cys, Ala-Gln, Ala-Tyr, Leu-Glu, Leu-Ser, Leu-Asp, Leu covalently bonded to a first amino acid selected from the ASn, Leu-Thr, Leu-Arg, Leu-Cys, Leu-Gln, Leu-Tyr, Phe group consisting of Ser, LyS, Glu, Asp, Ala, Leu, Phe, Val, Glu, Phe-Ser, Phe-Asp, Phe-ASn, Phe-Thr, Phe-Arg, Phe Gly, Tyr, Pro, and Thr, and a second amino acid or peptide Cys, Phe-Gln, Phe-Tyr, Phe-Phe-Ile, Phe-Phe-Leu, Phe-Phe covalently bonded to the first amino acid, wherein an optional Lys-Phe-Phe, Val-Glu, Val-Ser, Val-Asp, Val-Asn, Val-Thr, linker attaches the first amino acid to the drug radical may be Val-Arg, Val-Cys, Val-Gln, Val-Tyr, Val-Val, Gly-Gly-Leu, utilized in connection with the present invention. Gly-Gly-Gly-Gly-Leu, Gly-Gly-Ile, Tyr-Tyr-Ile, Tyr-Tyr In one embodiment, a compound comprising a drug radical Phe-Phe-Ile, Tyr-Tyr-Val, Tyr-Tyr-Phe, Tyr-Tyr-Lys-Tyr-Tyr, covalently bonded to a peptide selected from the group con Asp-Asp-Ile, Pro-Pro-Leu, Pro-Pro-Ile, and Thr-Thr-Val. sisting of Lys-Lys, Lys-Lys-Val. (Lys-Lys-Gly-Gly), (1)- 25 wherein an optional linker attaches the first amino acid to the Lys-(d)-Lys-Leu, Glu-Glu, Glu-Glu-Phe-Phe-Phe-Ile, Glu drug radical may be utilized in connection with the present Glu-Phe-Phe-Phe, Glu-Glu-Phe-Phe-Ile, Glu-Glu-Val, Asp invention. Asp, Asp-Asp-Asp, Asp-Asp-Glu, Asp-Asp-Ser, Asp-Asp In one embodiment, a compound comprising a drug LyS, Asp-Asp-Cys, Asp-Asp-Val. Ala-Ser, Ala-Cys, Leu-Glu, attached to a first amino acid via a carboxyl group of the first Leu-Ser, Leu-Asp, Leu-ASn, Leu-Thr, Leu-Arg, Leu-Cys, 30 amino acid, which forms an ester linkage between the first Leu-Gln, Leu-Tyr, Val-Glu, Val-Ser, Val-Asp, Val-Asn, Val amino acid and the drug, and a second amino acid or peptide Thr, Val-Arg, Val-Cys, Val-Gln, Val-Tyr, Val-Val, Gly-Gly attached to the first amino acid through the terminal carboxyl Leu, Gly-Gly-Gly-Gly-Leu, Gly-Gly-Ile, Asp-Asp-Ile, Pro group of the second amino acid or peptide, wherein the drug Pro-Leu, Pro-Pro-Ile, and Thr-Thr-Val, wherein an optional is Susceptible to release from the compound in therapeutically linker attaches the first amino acid to the drug radical may be 35 significant amounts through enzymatic cleavage, and not utilized in connection with the present invention. released from the compound in therapeutically significant In one embodiment, a compound comprising a drug radical amounts absent enzymatic action may be utilized in connec covalently bonded to a peptide selected from the group con tion with the present invention. sisting of Ser-Ser, PolySer, Lys, Glu-Glu, Asp-Asp, Asp-Asp In one embodiment, a compound comprising a drug Asp, Asp-Asp-Glu, Asp-Asp-Ser, Asp-Asp-Lys, Asp-Asp 40 attached to a first amino acid via a carboxyl group of the first Cys, Ala-Glu, Ala-Ser, Ala-Asp, Ala-ASn, Ala-Thr, Ala-Arg, amino acid, which forms an ester linkage between the first Ala-Cys, Ala-Gln, Ala-Tyr, Leuglu, Leu-Ser, Leu-Asp, Leu amino acid and the drug, and a second amino acid or peptide ASn, Leu-Thr, Leu-Arg, Leu-Cys, Leu-Gln, Leu-Tyr, Phe attached to the first amino acid through the terminal carboxyl Glu, Phe-Ser, Phe-Asp, Phe-ASn, Phe-Thr, Phe-Arg, Phe group of the second amino acid or peptide, wherein the drug Cys, Phe-Gln, Phe-Tyr, Val-Glu, Val-Ser, Val-Asp, Val-Asn. 45 is susceptible to release from the compound through enzy Val-Thr, Val-Arg, Val-Cys, Val-Gln, and Val-Tyr, wherein an matic cleavage may be utilized in connection with the present optional linker attaches the first amino acid to the drug radical invention. may be utilized in connection with the present invention. In one embodiment, a compound comprising a drug In one embodiment, a compound comprising a drug radical covalently bonded to a first amino acid via a carboxyl group covalently bonded to a peptide selected from the group con 50 of the first amino acid, which forms an ester linkage between sisting of Pro-Pro-Leu, Pro-Pro-He, Val-Val. Lys-Lys, Gly the first amino acid and the drug, and a second amino acid or Gly-Ile, Phe-Phe-He, Phe-Phe-Leu, Thr-Thr-Val, Tyr-Tyr peptide attached to the first amino acid through the terminal Val, Tyr-Tyr-Phe, Glu-Glu-Val. Asp-Asp-Val. Lys-Lys-Val. carboxyl group of the second amino acid or peptide thereby Glu-Glu-Phe-Phe-Ile, Glu-Glu-Phe-Phe-Phe, Tyr-Tyr-Ile, forming a peptide bond with an NH group of the first Asp-Asp-He, Tyr-Tyr-Phe-Phe-Ile, Tyr-Tyr-Lys-Tyr-Tyr, 55 amino acid, wherein the drug is susceptible to release from Phe-Phe-Lys-Phe-Phe, Glu-Glu-Phe-Phe-Ile, (Lys-Lys-Gly the compound through enzymatic cleavage may be utilized in Gly), and (1)-Lys-(d)-Lys-Leu, wherein an optional linker connection with the present invention. attaches the first amino acid to the drug radical may be uti In one embodiment, a compound comprising an active lized in connection with the present invention. agent covalently bonded to a natural or synthetic amino acid In one embodiment, a compound comprising a drug radical 60 or peptide through a linker may be utilized in connection with covalently bonded to a peptide selected from the group con the present invention. The linker may comprise a small linear sisting of Ser-Ser, PolySer, Glu-Glu, Asp-Asp, Asp-Asp-Asp, or cyclic molecule containing 2-6 atoms with one or more Asp-Asp-Glu, Asp-Asp-Ser, Asp-Asp-Lys, Asp-Asp-Cys, heteroatoms and one or more functional groups. The func Ala-Glu, Ala-Ser, Ala-Asp, Ala-ASn, Ala-Thr, Ala-Arg, Ala tional groups may be selected from amines, amides, alcohols Cys, Ala-Gln, Ala-Tyr, Leu-Glu, Leu-Ser, Leu-Asp, Leu 65 or acids. ASn, Leu-Thr, Leu-Arg, Leu-Cys, Leu-Gln, Leu-Tyr, Phe Compositions of the invention comprise four essential Glu, Phe-Ser, Phe-Asp, Phe-ASn, Phe-Thr, Phe-Arg, Phe types of attachment. These types of attachment are termed: