USOO9446.134B2 (12) United States Patent (10) Patent No.: US 9,446,134 B2 Maggio (45) Date of Patent: *Sep. 20, 2016 (54) STABILIZING ALKYLGLYCOSIDE 5, 198420 A 3, 1993 Donahoe et al. COMPOSITIONS AND METHODS THEREOF 5,369,095 A 11/1994 Kee 5,556,757 A 9/1996 Alstyne et al. (71) Applicant: Aegis Therapeutics, LLC, San Diego, 5,556,940 A 9, 1996 Willick et al. 5,661,130 A 8, 1997 Meezan CA (US) 6,524,557 B1 2/2003 Backstrom et al. (72) Inventor: Edward T. Maggio, San Diego, CA 6,551,578 B2 4/2003 Adjei et al. (US) 7,425,542 B2 9/2008 Maggio 7.998,927 B2 8/2011 Maggio (73) Assignee: Aegis Therapeutics, LLC, San Diego, 8,133,863 B2 * 3/2012 Maggio ........................ 514,111 CA (US) 8,226,949 B2 7/2012 Maggio 2002/0110524 A1 8, 2002 Cowan et al. (*) Notice: Subject to any disclaimer, the term of this 2004/0209814 A1 10/2004 Nauck patent is extended or adjusted under 35 2005/0215475 A1 9/2005 Ong et al. U.S.C. 154(b) by 0 days. 2005/0276843 A1 12/2005 Quay et al. 2006, 0046962 A1 3/2006 Meezan et al. This patent is Subject to a terminal dis 2006, OO74025 A1 4/2006 Quay et al. claimer. 2006/00846.04 A1 4/2006 Kitaura et al. 2006,0183674 A1 8, 2006 Brand (21) Appl. No.: 14/494,990 2007/011 1938 A1 5/2007 Pert 2008. O194461 A1 8/2008 Maggio (22) Filed: Sep. 24, 2014 2008/0268032 A1 10/2008 Maggio (65) Prior Publication Data 2009/0047347 A1 2/2009 Maggio 2009. O156478 A1 6/2009 Lau et al. US 2015/0216980 A1 Aug. 6, 2015 2009/0326193 A1 12/2009 Maggio Related U.S. Application Data FOREIGN PATENT DOCUMENTS (63) Continuation of application No. 13/544,851, filed on WO WO 2006/025882 A3 3, 2006 Jul. 9, 2012, now Pat. No. 8,846,044, which is a WO WO 2006/051110 A3 5, 2006 continuation of application No. 12/618,558, filed on Nov. 13, 2009, now Pat. No. 8,226,949, which is a continuation-in-part of application No. 12/119,378, OTHER PUBLICATIONS filed on May 12, 2008, now Pat. No. 7.998,927, which Brown and Schonbrunn, "Affinity Purification of a Somatostatin is a continuation-in-part of application No. Receptor-G-Protein Complex demonstrates Specificity in Receptor 12/050,038, filed on Mar. 17, 2008, now Pat. No. G-Protein Coupling.” J. Biol. Chem. (1993), 268(9):6668-6676. The 8,084,022, which is a continuation-in-part of American Society for Biochemistry and Molecular Biology, Inc. application No. 11/474,055, filed on Jun. 23, 2006, Hovgaard et al., "Stabilization of Insulin by Alkylmaltosides. A. now Pat. No. 7,425,542. Spectroscopic Evaluation.” Int. J. Pharm. (1996), 132: 107-113, (51) Int. Cl. Elsevier Science B.V. A6 IK 38/08 (2006.01) Hovgaard et al., "Stabilization of Insulin by Alkylmaltosides. B. A6 IK 47/26 (2006.01) Oral Absorption in vivo in Rats.” Int. J. Pharm. (1996), 132:115 A6 IK 9/00 (2006.01) 121, Elsevier Science B.V. A 6LX 9/27 (2006.01) Mitrano and Newton, “Factors Affectina Insulin Adherence to Type A 6LX 9/19 (2006.01) I Glass Bottles”, Am. J. Hosp. Pharm. (1982), 39(9): 1491-1495. Van Der Lubben et al., “Chitosan and its Derivatives in Mucosal A6 IK3I/70 (2006.01) Drug and Vaccine Delivery.” Eur.J. Pharm, Sci. (2001) 14:201-207, A6 IK 45/06 (2006.01) Elsevier. A6 IK 47/10 (2006.01) Ahsan et al., “Mutual inhibition of the insulin absorption-enhancing A6 IK 47/8 (2006.01) properties of dodecylmaitoside and dimethyl-beta-cyclodextrin fol A6 IK 47/36 (2006.01) lowing nasal administration’. Pharm Res., 18(5):608-14 (2001). (52) U.S. Cl. (Continued) CPC ............. A61K 47/26 (2013.01); A61K 9/0043 (2013.01); A61K 9/127 (2013.01); A61K 9/19 (2013.01); A61 K3I/70 (2013.01); A61K 38/08 Primary Examiner — Jeffrey E. Russel (2013.01); A61K 45/06 (2013.01); A61K 47/10 (2013.01); A61K 47/186 (2013.01); A61 K (74) Attorney, Agent, or Firm — DLA Piper LLP (US) 47/36 (2013.01) (58) Field of Classification Search (57) ABSTRACT None See application file for complete search history. The present invention relates to alkylglycoside-containing compositions and methods for increasing the stability, (56) References Cited reducing the aggregation and immunogenicity, increasing the biological activity, and reducing or preventing fibrillar U.S. PATENT DOCUMENTS formation of a peptide, polypeptide, or variant thereof, for 4,130,709 A 12/1978 Nagarajan example octreotide or variant thereof. 4,440,675 A 4, 1984 Braude 4476,116 A 10, 1984 Anik 5, 122,187 A 6, 1992 Schwarz et al. 25 Claims, 15 Drawing Sheets US 9,446,134 B2 Page 2 (56) References Cited European Search Report (ESR) from application No. EP 1079 2685., completed Jul. 8, 2013. OTHER PUBLICATIONS Tachibana, Hiroshi et al.: “Differentiation of Entamoeba histolytica Vanaken et al., “Alkyl glycoside detergents: synthesis and applica- fkDE. diIspar C.ilitated al P by C. lonalt santibodi i 9 t3inst tions to the study of membrane proteins”. Methods Enzymol. -kDa surface antigen”; Parasitol Res, May 1, , pp. ty. 125:27-35 (1986). European Search Report issued on Mar. 21, 2016, regarding EP 12 Maggio, E.T., “Intravail: highly effective intranasal delivery of 165 052.7. peptide and protein drugs'. Expert Opin. Drug Deliv. 3(4):529-39 (2006). * cited by examiner U.S. Patent Sep. 20, 2016 Sheet 1 of 15 US 9,446,134 B2 Background Information Figure 1 U.S. Patent Sep. 20, 2016 Sheet 2 of 15 US 9,446,134 B2 insulin pH 6.5 1200 1OOO -- Control s 8OO -- 0.062% A 0.125% A 600 - x - O.25.0% A 5. 400 -k- OO62 B -o-O.125% B 200 --O.250% B O O 1 2 6 9 13 16 2O Days 37C Figure 2 U.S. Patent Sep. 20, 2016 Sheet 3 of 15 US 9,446,134 B2 insulin pH 7.4 -0- Control -- 0.062% A c -- 0.125% A s -3%a O.250% A wd -- 0.062% B e -O-O.125% B -- 0.250% B Days 37C Figure 3 U.S. Patent Sep. 20, 2016 Sheet 4 of 15 US 9,446,134 B2 hCGh pH 6.5 Intravail 0.125% 10ul sample -0- Control --Intravail 7 10 13 16 20 Days hGH pH 6.5 intravail 0.124% 5ul Sample 1 5O 1 OO -0- Control 5O -- intravail O O 1 2 3 7 10 13 16 20 Figure 4 U.S. Patent Sep. 20, 2016 Sheet 5 Of 15 US 9,446,134 B2 Comparison of Time Dependent Effect of Aggregation of Peptide T (DAPTA) Stored at 4 Degrees for Prolong Period of Time (Lot 036299) s -0-6 h 14 degree s w -- 1 week s -A-2 weeks - orio 3 weeks w -- 4 weeks -O-2 months -10 -9 Peptide T (DAPTA) LogM Figure 5 U.S. Patent Sep. 20, 2016 Sheet 6 of 15 US 9,446,134 B2 DAPTA Formulation (0.5 mg/ml). Inhibition of HIV-1 (ADA) Infection of Macrophages: Various Treatments for Stability A3+ DAPTA/ not TFE Tx, aged 14d A3+ DAPTAWTFE Tx, aged 14d DAPTA/TFE Tx, aged i4d B3+ DAPTA/ no TFE Tx, aged 14d B3 + DAPTA/TFE, aged 14d DAPTA aged 14d DAPTA aged 7d Virus only 25 50 75 1OO 1.25 Reverse Transcriptase Activity Percent of Virus Only Control by 0.1nM DAPTA Figure 6 U.S. Patent Sep. 20, 2016 Sheet 7 Of 15 US 9,446,134 B2 insulin pH 7.6 1 O OO s (d t w c o - es s 1. 11 14 20 27 39 45 54 60 67 75 82 88 Days at 37C, 150 RPM -- Control -A-0.1% dodecyl maltoside (less than 10% B anomer) --X -- 0.2% dodecyl maltoside (less than 10% B anomer) - - -X -- 0.1% dodecyl maltoside (greater than 99% B anomer) -o-0.2% dodecyl maltoside (greater than 99% 8 anomer) Figure 7 U.S. Patent Sep. 20, 2016 Sheet 8 of 15 US 9,446,134 B2 Ostabolin C, pH 3.5 40 deg. C, 150 RPM 120 OO 8 O 60 40 20 -20 O.O 2.0 4.0 6.0 8.0 0.0 2.0 14.0 Figure 8 U.S. Patent Sep. 20, 2016 Sheet 9 Of 15 US 9,446,134 B2 Ostabolin C, pH 5.0 40 deg. C, 150 RPM 1000 8 O O 0.0 2.0 4.0 6.0 8.0 10.0 2.0 14.0 Figure 9 U.S. Patent Sep. 20, 2016 Sheet 10 of 15 US 9,446,134 B2 PTH 1-34, pH 3.0, 40 deg. C 0. 0 O 800 6 0 0 - 400 200 rer Figure 10 U.S. Patent Sep. 20, 2016 Sheet 11 of 15 US 9.446,134 B2 PTH 1-34, pH 5.0, 40 deg. C 200 - Figure 11 U.S. Patent Sep. 20, 2016 Sheet 12 of 15 US 9.446,134 B2 1200 PTH 1-34, pH 5.0 at 37 deg. C 1000 800 600 400 200 -200 Day Figure 12 U.S. Patent Sep. 20, 2016 Sheet 13 of 15 US 9,446,134 B2 800 700 600 500 400 300 200 100 |--|--|-- |--|--|-- |-|--|-- 0.00twE.