US009394339B2  c12)  United States Patent  Anderson  (54)  STABILIZED FORMULATIONS OF PEPTIDES  AND PROTEINS PROTECTING AGAINST  NON-ENZYMATIC DEGRADATION  (IO)  Patent No.:  US 9,394,339 B2  (45)  Date of Patent:  Jul. 19, 2016  (71)  Applicant:  Lyotropic Therapeutics, Inc.,  Ashland,  VA (US)  (72)  Inventor:  David Anderson,  Ashland, VA (US)  (73)  Assignee:  Lyotropic Therapeutics, Inc.,  Ashland,  VA (US)  ( *)  Notice:  Subject to any disclaimer, the term ofthis  patent is extended or adjusted under 35  U.S.C. 154(b) by  O  days.  (21)  Appl. No.:  14/477,925  (22)  Filed:  Sep. 5, 2014  (63)  Continuation of application No. 12/809,206, filed as  application No. PCT/US2008/087611 on Dec. 19,  2008, now Pat. No. 8,858,927.  (60)  Provisional application No. 61/122,789, filed on Dec.  16, 2008, provisional application No. 61/015,993,  filed on Dec. 21, 2007.  (51)  Int. Cl.  (52)  U.S. Cl.  CPC .................  C07K 7106  (2013.01);  A61K 9/0019  (2013.01);  A61K 9/0048  (2013.01);  A61K 9/19  (2013.01);  A61K  47120  (2013.01);  A61K  47126  (2013.01)  (58)  Field of Classification Search  None  See application file for complete search history.  (65)  Prior Publication Data  OTHER PUBLICATIONS  FOREIGN PATENT DOCUMENTS  (56)  References Cited  (57)  ABSTRACT  5 Claims, 1 Drawing Sheet  U.S. PATENT DOCUMENTS  Related U.S. Application Data  US 2014/0378655 Al  Dec. 25, 2014  Potter, S., et al. "In vitro aging of calmodulin gnerates isoaspartate at  multiple Asn-Gly and Asp-Gly sites in calcium-binding domans II,  III, and IV"; Protein Science (1993), 2, pp. 1648-1663.  Nilsson, M., et al., "Low levels of asparagine deamidation can have a  dramatic effect on aggregation of amyloidogenic peptides: Implica› tions for the study of amyloid formation"; Protein Science (2002) 11,  pp. 342-349.  Perna, A., et al., "Plasma protein aspartyl damage is increased in  hemodialysis patents: Studies on causes and consequences"; J. Am  Soc. Pephrol. (2004)15, pp. 2747-2754.  Paleari, R., et al., Posttranslational deamidation of proteins: the case  of Hemoglobin j sardegna [ alpha50(CD8)His> Asn> Asp]; Clinical  Chemistry, 45:1, Molecular Diagnostics and Genetics (1999) pp.  21-28.  Robinson, N., et al., "Predictin of primary structure deamidationrates  of asparaginyl and glutaminyl peptides through steric and catalytic  effects"; J. Peptide Res., 2004, 63, 437-448.  Cloos, P., et al., "Post-translational modifications of proteins: impli› cations for aging,  antigen recognition,  and autoimmunity";  Biogerontology 5: (2004); pp. 139-158.  Wakankar, A., et al., "Formulation considerations for proteins sus› ceptible to asparagine deamidatino and aspartate isomerization"; J. of  Pharm Sciences, vol. 95, No. 11,(2006) pp. 2321-2336.  Cournoyer, et al., "Analysis of Deamidation in Proteins", Compre› hensive Analytical Chemistry, 52, 37 5-410 (2009).  Wright, T.H., "Nonenzymatic Deamidation of Asparaginyl and  Glutaminyl Residues in Proteins", Crit. Rev. Biochem. Mo!. Biol. 26,  1-52 (1991).  * cited by examiner  Primary Examiner  - Elly-Gerald Stoica  (74)  Attorney, Agent, or Firm  - Whitham, Curtis & Cook,  P.C.  Stabilization of water-containing solutions or lyophilizates of  proteins and peptide against non-enzymatic deamidation deg› radation reactions at asparaginyl or glutaminyl residues is  achieved using organic anions, such as saccharin, benzene› sulfonic acid, gentisic acid or N-acetyltryptophan which have  a pKa within the range of0.5 to 3.5.  3,876,775 A *  4/1975 Izaka  A61K 38/08  514/15.2  5,238,917 A *  8/1993 Fujii  A61K 9/0034  424/430  A61K9/00  A61K9/19  A61K  47120  A61K  47126  A61K 45100  A61K 38121  A61K 39/395  C07K 7106  (2006.01)  (2006.01)  (2006.01)  (2006.01)  (2006.01)  (2006.01)  (2006.01)  (2006.01)  CN  1565413 Al *  1/2005  A61K 7/16  5,569,602 A *  10/1996 Lunardi-Iskandar Cl2N 5/0693  435/366  6,013,773 A  1/2000 Kobayashi et al.  6,274,553 Bl  8/2001  Furuya et al.  2003/0092622 Al  5/2003 Sato et al.  2006/0013936 Al *  1/2006 Miskovsky  A23C 9/1315  426/580  2006/0234989 Al  10/2006 Anderson  2007/0003502 Al*  1/2007 Tanabe.  A61K 8/0212  424/70.13 