USOO8133863B2 (12) United States Patent (10) Patent No.: US 8,133,863 B2 Maggio (45) Date of Patent: Mar. 13, 2012 (54) STABILIZING ALKYLGLYCOSIDE 3.5. A g E. S. et al COMPOSITIONS AND METHODS THEREOF 5,369,095 A 1 1/1994 KeeOnahoe et al. et al. 5,556,757 A 9, 1996 A1 tal. (75) Inventor: Edward T. Maggio, San Diego, CA 5,556,940 A 9, 1996 WRC (US) 6,524,557 B1 2/2003 Backstrom et al. 6,551.578 B2 * 4/2003 Adjei et al. ..................... 424/45 (73) Assignee: Aegis Therapeutics, LLC, San Diego, 2002fO1105247,425,542 A1B2 9/20088, 2002 CowanMaggio et al. CA (US) 2004/0209814 A1 10, 2004 Nauck et al. 2005/0215475 A1 9/2005 Ong et al. (*) Notice: Subject to any disclaimer, the term of this 2005/0276843 Al 12/2005 Quay et al. patent is extended or adjusted under 35 2006/0046962 A1 3/2006 Meezan et al. .................. 514/12 2006, OO74025 A1 4/2006 Quay et al. U.S.C. 154(b) by 443 days. 2006/0183674 A1 8/2006 Brand et al. 21) Appl. No.: 12/360,758 2007/01 11938 A1 5, 2007 Pert et al. (21) Appl. No.: 9 (Continued) (22) Filed: Jan. 27, 2009 FOREIGN PATENT DOCUMENTS US 2009/0197816A1 Aug. 6, 2009 (Continued) OTHER PUBLICATIONS Related U.S. Application Data Brown et al. Affinity Purification of a Somatostatin Receptor-G- (60) Division of application No. 12/119,378, filed on May protein Complex... The Journal of Biological Chemistry. Mar. 25. 12, 2008, which is a continuation-in-part of 1993, Vol. 268, pp. 6668-6676. application No. 12/050,038, filed on Mar. 17, 2008 Hovgaardet al., "Stabilization of insulin by alkylmaltosides. A. Spec which is a continuation-in- art of a lication No. troscopic evaluation'. International Journal of Pharmaceutics, p pp 132(1-2): 107-113 (1996). 1 1/474,055, filed on Jun. 23, 2006, now Pat. No. 7,425,542. (Continued) 51) Int. C Primary Examiner — Jeffrey E. Russel (51) 38/08 (2006.01) (74) Attorney, Agent, or Firm — DLA Piper LLP (US) A6 IK38/2 (2006.01) (57) ABSTRACT A6 IK 47/26 (2006.01) The present invention relates to alkylglycoside-containing (52) U.S. Cl. ...................................... 514/11.1: 514/21.1 compositions and methods for increasing the stability, reduc (58) Field of Classification Search ........................ None ing the aggregation and immunogenicity, increasing the bio See application file for complete search history. logical activity, and reducing or preventing fibrillar formation of a peptide, polypeptide, or variant thereof, for example (56) References Cited amylin, a monoclonal antibody, insulin, Peptide T or analog thereof, gastrin, gastrir releasing peptides, gastrin releasing U.S. PATENT DOCUMENTS peptide-like (GRP) proteins, epidermal growth factor orana 4,130,709 A 12/1978 Nagarajan log thereof. 4,440,675 A 4, 1984 Braude 37 Claims, 7 Drawing Sheets Background Information Fibri Oxis Four sheets US 8,133,863 B2 Page 2 U.S. PATENT DOCUMENTS OTHER PUBLICATIONS 2008. O194461 A1 8, 2008 Maggio 2008/0268032 A1 10, 2008 Maggio Hovgaard et al., “Stabilization of insulin by alkylmaltosides. B. Oral 2009/0047347 A1 2, 2009 Maggio absorption in vivo in rats'. International Journal of Pharmaceutics, 2009. O156478 A1 6, 2009 Lau et al. 132(1-2): 115-121 (1996). 2009/0326193 A1 12, 2009 Maggio et al. van der Lubben et al., "Chitosan and its derivatives in mucosal drug and vaccine delivery'. Eur, J. Pharm. Sci., 14(3):201-207 (2001). FOREIGN PATENT DOCUMENTS Mitrano & Newton, “Factors affecting insulin adherence to type I WO WO 2006/025882 A3 3, 2006 glass bottles”, Am. J. Hosp. Pharm., 39(9): 1491-5 (1982). WO WO 2006/051110 A2 5, 2006 WO WO 2006/051110 A3 5, 2006 * cited by examiner U.S. Patent Mar. 13, 2012 Sheet 1 of 7 US 8,133,863 B2 Background Information Fig. 1 U.S. Patent Mar. 13, 2012 Sheet 2 of 7 US 8,133,863 B2 T-contro" 8-8282% A 3.125% A 3 8 -: - & 3.25.0% A k-3.08.2% 8 -X-3:23%. 8: .'-' ' '.' '.' '.'''''''-' ' ' ' ' ' ' ' ' ' ' '-'-'-'s' - - - - - - - - - 3.35.3% 8. U.S. Patent Mar. 13, 2012 Sheet 3 of 7 US 8,133,863 B2 -8-0.062% A -- 0.25% A -- 0,250% A -%-0.082% 8. -8-0.25% B i -- 0.250% B U.S. Patent Mar. 13, 2012 Sheet 4 of 7 US 8,133,863 B2 Gh p 6.5 traya .25% sapie '''''''x''-W-swaxwww.axwww.www.W.W ------------------------------a .....-------as-a-was--------- 3-ph S.S. travai (, 24% St. sapie 188 go-cr; U.S. Patent Mar. 13, 2012 Sheet 5 of 7 US 8,133,863 B2 Coaparisor of irs begside: Effect of Aggregation of peptide {A-3A Stored at 4 Degrees for Preierg period of inte (lot 336238 -8-6-----. .hia - - - - - - degree:- - - - - - : -8-3 week :*:- 2 wa&ks 3 weaks i-x-4 weaks Pepticis ; (383&; lag 8 Fig. 5 U.S. Patent Mar. 13, 2012 Sheet 6 of 7 US 8,133,863 B2 n 3&A Ferraiation; (3.8 rigfa risixties of 3W-3 &A seceign of 8-acre ages: variots reatests for Staixity A3+ APA, not FE Ex, aged A3 - APA/FE x, aged 34. A 8: Afif x, aged : 83+ CAPTA, no TFE Tx, 33 - DAPAf:FE, aged CAPTA aged 34d x BAPIA aged 7d x : wifts ory E8 SE AS s &ewerse raiseritese &xivity *erce of virts City Cerro: y 8.38: 888 & Fig. 6 U.S. Patent Mar. 13, 2012 Sheet 7 of 7 US 8,133,863 B2 {{3} 800 63 f f : ji 40G ? f 200 X. --- 3 : S. 7 & 28, 2 39 43 54; 8: 8? 73 82 88 Rays at 37C, 353 RP8; -e-contro --0.1% dodecy maitoside (less than 10% 8 anomer) - x - -3.2% dodecy; naitoside (less than 30% Sanomer) x . 4% dodecy: iaitoside (greater {ha : 98% 8 artorner) -x- 3.2% 8ociecy: 3:a::side great &f that 88% 3 artorie US 8,133,863 B2 1. 2 STABILIZING ALKYLGLYCOSDE Many products are only effective when delivered by injec COMPOSITIONS AND METHODS THEREOF tion in relatively high concentration. Preventing aggregation has become a major issue for pharmaceutical formulators CROSS REFERENCE TO RELATED since the trend toward high-concentration solutions increases APPLICATION(S) the likelihood of protein-protein interactions favoring aggre gation. Thus, protein aggregation may impact biological This application is a divisional of U.S. application Ser. No. product process yield and potency. Since aggregation is fre 12/119,378, filed May 12, 2008, currently pending: which is quently mediated by higher temperatures, protein therapeu a continuation-in-part of U.S. application Ser. No. 12/050, tics require certain so-called “Cold Chain’ handling require 10 ments to guarantee a continuous chain of refrigerated 038 filed Mar. 17, 2008, currently pending; which is a con temperatures during shipping and storage (DePalma Jan. 15. tinuation-in-part of U.S. application Ser. No. 1 1/474,055 filed 2006). Cold chain requirements significantly increase the Jun. 23, 2006 now U.S. Pat. No. 7,425,542, now issued. The cost of storing and transporting drugs. The present invention entire content of each of the prior applications is incorporated mitigates and, in some cases, may eliminate the need for strict herein by reference. 15 cold-chain maintenance. Over the last five years, the FDA and other regulatory FIELD OF THE INVENTION agencies have increased their scrutiny of aggregation events, and thus biopharmaceutical companies have increased their The present invention relates generally to compositions efforts to understand them. Of particular concernis the induc and methods thereof that increase stability, reduce aggrega tion of unwanted immunogenicity. The immunogenicity of a tion and immunogenicity, increase biological activity, and self-associating peptide can be influenced by the formation of reduce or prevent fibrillar formation of peptides or proteins in aggregates formed as a result of non-covalent intermolecular therapeutically useful formulations, and specifically, to com interactions. For example, interferon has been shown to positions having at least one peptide or protein drug and at aggregate resulting in an antibody response (Hermeling et al. least one alkylglycoside or saccharide alkyl ester Surfactant. 25 2006). The antibody response to erythropoietin has been shown to produce “pure red cell aplasia” in a number of BACKGROUND INFORMATION patients receiving recombinant EPO, (Casadevallet al. 2002) which is potentially a life threatening side effect of EPO Proteins undergo numerous physical and chemical changes therapy. Insulin is well known to lose activity rapidly as a that affect potency and safety. Among these are aggregation, 30 result of protein aggregation upon agitation at temperatures which includes dimerization, trimerization, and higher-order above those found upon refrigerated Storage (Pezron et al. aggregates, plus crystallization and precipitation. Aggrega 2002; Sluzky et al. 1991). Aggregation of recombinant AAV2 tion is rapidly emerging as a key issue underlying multiple results in reduced yield during purification and has deleteri deleterious effects for peptide or protein-based therapeutics, ous effects on immunogenicity following in vivo administra including loss of efficacy, altered pharmacokinetics, reduced 35 tion (Wright 2005). Monoclonal antibody based therapeutics stability or product shelf life, and induction of unwanted have also been shown to be subject to inactivation as a result immunogenicity. In addition, bioavailability and pharmaco of protein aggregation (King et al. 2002). The number of kinetics of a self-associating peptide can be influenced by monoclonal antibodies in human clinical trials has been on aggregate size and the ease of disruption of the non-covalent the rise. Often monoclonal antibodies require relatively high intermolecular interactions at the Subcutaneous site.