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# c12) United States Patent Anderson 

# (54) STABILIZED FORMULATIONS OF PEPTIDES AND PROTEINS PROTECTING AGAINST NON-ENZYMATIC DEGRADATION 

(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 

## (65) Prior Publication Data 

US 2014/0378655 Al Dec. 25, 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. 

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(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. 

## (56) References Cited 

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## Related U.S. Application Data 

## U.S. PATENT DOCUMENTS 

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# (IO) Patent No.: US 9,394,339 B2 

# (45) Date of Patent: Jul. 19, 2016 

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CN 1565413 Al * 1/2005 A61K 7/16 

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. 

## (57) ABSTRACT 

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. 

US009394339B2 

## FOREIGN PATENT DOCUMENTS 

## OTHER PUBLICATIONS 

## 5 Claims, 1 Drawing Sheet 

