IP Library › Granted Patent US 12,365,768
Granted Patent B2
US 12,365,768 · App. 17/676,675 · Granted Jul 22, 2025

Method for deacetylation of biopolymers

Inventors: Johan Olsson (Bromma, SE); Craig Steven Harris (Biot, FR)
Assignee: Galderma Holding SA
C08J3/075A61K8/042A61K8/73A61K8/735A61K9/0019A61Q19/00C07C209/62C07C213/00C07C269/06C07F7/083C08B37/0063C08B37/0069C08B37/0072C08J3/24C08J7/14C08K5/09C08L5/00C08L5/08C07C2603/18C08J2305/00C08J2305/08
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Quick Facts
Patent No.
US 12,365,768
App. No.
17/676,675
Granted
Jul 22, 2025
Kind
B2
Abstract

A method for at least partial deacetylation of a biopolymer comprising acetyl groups, including: a1) providing a biopolymer including acetyl groups; a2) reacting the biopolymer including acetyl groups with hydroxylamine (NH 2 OH) or a salt thereof at a temperature of 100° C. or less for 2-200 hours to form an at least partially deacetylated biopolymer; and a3) recovering the at least partially deacetylated biopolymer.

Claims (26)

1. A method of preparing a hydrogel product comprising crosslinked glycosaminoglycan molecules, the method comprising:

(a) obtaining a solution comprising an at least first glycosaminoglycan that is a partially deacetylated glycosaminoglycan with a degree of acetylation between 50% and 99% and a second glycosaminoglycan, wherein the second glycosaminoglycan is different from the at least partially deacetylated glycosaminoglycan;

(b) activating carboxyl groups on the at least partially deacetylated glycosaminoglycan and/or the second glycosaminoglycan with a coupling agent, to form activated glycosaminoglycans;

(c) crosslinking the activated glycosaminoglycans, solely with amide bonds, using a crosslinker comprising at least two amine groups, to provide crosslinked glycosaminoglycans; and

(d) acylating residual amine groups of the crosslinked glycosaminoglycans obtained in (c) to form acylated crosslinked glycosaminoglycans; and

(e) subjecting the crosslinked glycosaminoglycans provided in (c) or (d) to alkaline treatment to hydrolyze ester crosslinks formed as byproducts during the amide crosslinking in (c),

wherein the first and second glycosaminoglycans are not the crosslinker, and wherein the crosslinker comprises a di-, tri-, or tetra-saccharide, oligosaccharide, or polysaccharide.

2. The method according to claim 1 , wherein the second glycosaminoglycan is at least partially deacetylated.

3. The method according to claim 1 , wherein at least 1% of N-acetyl groups of the first glycosaminoglycan have been converted to free amine groups.

4. The method according to claim 2 , wherein at least 1% of N-acetyl groups of the second glycosaminoglycan have been converted to free amine groups.

5. The method according to claim 1 , wherein the second glycosaminoglycan is not deacetylated.

6. The method according to claim 1 , wherein the first and second glycosaminoglycans are selected from hyaluronic acid, chondroitin, chondroitin sulphate, heparin sulphate, heparosan, heparin, dermatan sulphate, and keratin sulphate.

7. The method according to claim 1 , wherein the first glycosaminoglycan and/or the second glycosaminoglycan is hyaluronic acid.

8. The method according to claim 1 , wherein the first glycosaminoglycan and the second glycosaminoglycan are hyaluronic acids.

9. The method according to claim 1 , wherein the crosslinker comprises a di, tri-, or tetra-saccharide, or combinations thereof.

10. The method according to claim 1 , wherein the crosslinker comprises a disaccharide.

11. The method according to claim 1 , wherein the crosslinker comprises diaminotrehalose (DATH).

12. The method according to claim 1 , wherein (b) and (c) occur simultaneously.

13. The method according to claim 1 , wherein (b) occurs prior to (c).

14. The method according to claim 1 , wherein the hydrogel product comprises a portion of glycosaminoglycans which are not crosslinked.

15. The method according to claim 1 , wherein the crosslinked glycosaminoglycan molecules comprise at least 80% by weight of the hydrogel product.

16. The method according to claim 1 , wherein the first glycosaminoglycan is a partially deacetylated glycosaminoglycan with a degree of acetylation between 93% and 99%.

17. The method according to claim 1 , wherein the second glycosaminoglycan obtained in a) is not altered or modified by addition or removal of functional groups.

18. The method according to claim 1 , wherein the first glycosaminoglycan has a weight average molecular weight of 0.5 MDa or more.

19. The method according to claim 1 , wherein the hydrogel product has a degree of amide crosslinking (CrDamide) of at least 0.5.

20. The method according to claim 1 , wherein the hydrogel product has a degree of amide crosslinking (CrDamide) of at least 0.9.

Priority Claims (4)
EP 15202944 · Dec 29, 2015 · regional
EP 16172225 · May 31, 2016 · regional
EP 16172241 · May 31, 2016 · regional
EP 16172254 · May 31, 2016 · regional
Continuity (2)
Continuation 16066746
Related Publication 20220177655A1 · Jun 9, 2022
References Cited (132)
US 5332812A · Nicolson et al. · 1994 [cited by applicant]
US 5731298A · Reinmuller · 1998 [cited by applicant]
US 6132750A · Perrier et al. · 2000 [cited by applicant]
US 6495314B1 · Kent et al. · 2002 [cited by applicant]
US 6703444B2 · Zhao et al. · 2004 [cited by applicant]
US 6831172B1 · Barbucci et al. · 2004 [cited by applicant]
US 8858999B2 · Giammona et al. · 2014 [cited by applicant]
US 8887243B2 · Thomson et al. · 2014 [cited by applicant]
US 10889894B2 · Hwung et al. · 2021 [cited by applicant]
US 11198765B2 · Olsson et al. · 2021 [cited by applicant]
US 11530301B2 · Mojarradi et al. · 2022 [cited by applicant]
US 20030093157A1 · Casares et al. · 2003 [cited by applicant]
US 20040014960A1 · Moon et al. · 2004 [cited by applicant]
US 20040072793A1 · Aeschlimann et al. · 2004 [cited by applicant]
US 20040219630A1 · Tsubouchi · 2004 [cited by applicant]
US 20060166928A1 · Moon et al. · 2006 [cited by applicant]
US 20070053987A1 · Bayer et al. · 2007 [cited by applicant]
US 20070065481A1 · Chudzik et al. · 2007 [cited by applicant]
US 20070066816A1 · Tsai et al. · 2007 [cited by applicant]
US 20090011045A1 · Mertin et al. · 2009 [cited by applicant]
US 20090247741A1 · Zhao · 2009 [cited by applicant]
US 20100136070A1 · Dobak et al. · 2010 [cited by applicant]
US 20100255068A1 · Stroumpoulis et al. · 2010 [cited by applicant]
US 20120231046A1 · Asius et al. · 2012 [cited by applicant]
US 20130085187A1 · Kim et al. · 2013 [cited by applicant]
US 20130203697A1 · Hashimoto et al. · 2013 [cited by applicant]
US 20130338352A1 · Yasugi et al. · 2013 [cited by applicant]
US 20140094568A1 · James et al. · 2014 [cited by applicant]
US 20150045573A1 · Cheng et al. · 2015 [cited by applicant]
US 20160106718A1 · Gupta · 2016 [cited by applicant]
US 20190016830A1 · Olsson et al. · 2019 [cited by applicant]
US 20190023812A1 · Mojarradi et al. · 2019 [cited by applicant]
US 20190023855A1 · Olsson et al. · 2019 [cited by applicant]
CN 1529716A · 2004 [cited by applicant]
CN 1538825A · 2004 [cited by applicant]
CN 1570128A · 2005 [cited by applicant]
CN 1694903A · 2005 [cited by applicant]
CN 102952281 · 2013 [cited by applicant]
CN 103038339A · 2013 [cited by applicant]
CN 103788222A · 2014 [cited by applicant]
CN 104194066A · 2014 [cited by applicant]
DE 4439575A1 · 1996 [cited by applicant]
EP 0224987A2 · 1987 [cited by applicant]
EP 0903152A2 · 1999 [cited by applicant]
EP 1837347A1 · 2007 [cited by applicant]
EP 2609924A1 · 2013 [cited by applicant]
EP 2682409A1 · 2014 [cited by applicant]
EP 2727597A1 · 2014 [cited by applicant]
EP 3020733A1 · 2016 [cited by applicant]
EP 3397651A1 · 2018 [cited by applicant]
JP S62265998A · 1987 [cited by applicant]
JP H11152234A · 1999 [cited by applicant]
JP 2002519481A · 2002 [cited by applicant]
JP 2004507588A · 2004 [cited by applicant]
JP 2004511588A · 2004 [cited by applicant]
JP 2006505633A · 2006 [cited by applicant]
JP 2009507103A · 2009 [cited by applicant]
JP 2014531433A · 2014 [cited by applicant]
JP 2015537078A · 2015 [cited by applicant]
RU 2230550C2 · 2004 [cited by applicant]
RU 2434633C2 · 2011 [cited by applicant]
RU 2448740C2 · 2012 [cited by applicant]
RU 2653729C2 · 2018 [cited by applicant]
WO WO9711958A1 · 1997 [cited by applicant]
WO WO0001733A1 · 2000 [cited by applicant]
WO WO0046252A1 · 2000 [cited by applicant]
WO WO0046253A1 · 2000 [cited by applicant]
WO WO0218450A1 · 2002 [cited by applicant]
WO WO0230990A1 · 2002 [cited by applicant]
WO WO02081739A2 · 2002 [cited by applicant]
WO WO02082078A2 · 2002 [cited by applicant]
WO WO2004011503A1 · 2004 [cited by applicant]
WO WO2004057008A1 · 2004 [cited by applicant]
WO WO2007026362A2 · 2007 [cited by applicant]
WO WO2013086024A2 · 2013 [cited by applicant]
WO WO2014072330A1 · 2014 [cited by applicant]
WO WO2015021092 · 2015 [cited by applicant]
WO WO2015034436 · 2015 [cited by applicant]
WO WO2015043757A1 · 2015 [cited by applicant]
WO WO2015181365A1 · 2015 [cited by applicant]
WO WO2015181369A1 · 2015 [cited by applicant]
WO WO2017114861 · 2017 [cited by applicant]
WO WO2017114864 · 2017 [cited by applicant]
WO WO2017114865 · 2017 [cited by applicant]
WO WO2017114867A1 · 2017 [cited by applicant]
WO WO2019001784A1 · 2019 [cited by applicant]
WO WO2019002368A1 · 2019 [cited by applicant]
WO WO2019002370A1 · 2019 [cited by applicant]
Jeon et al., “Mechanical properties and degradation behaviors of hyaluronic acid hydrogels cross-linked at various cross-linking densities”, Apr. 12, 2007, Carbohydrate Polymers, vol. 70, pp. 251-257. [cited by applicant]
U.S. Appl. No. 10/105,197-B1, filed Oct. 23, 2018, Colon, Fernando. [cited by applicant]
International Search Report and Written Opinion on PCT Appl. Ser. No. PCT/IB2020/061336 dated Mar. 1, 2021 (9 pages). [cited by applicant]
International Search Report and Written Opinion on PCT PCT/IB2020/060066 dated Jan. 12, 2021 (14 pages). [cited by applicant]
A Tsigos I et al: “Chitin deacetylases: new, versatile tools in biotechnology”, Trends in Biotechnology, Elsevier Publications, Cambridge, GB, vol. 18, No. 7, Jul. 1, 2000 (Jul. 1, 2000), pp. 305-312, XP004908536, ISSN:… [cited by applicant]
Babasola, Oladunni, et al., “Chemically Modified N-Acylated Hyaluronan Fragments Modulate Proinflammatory Cytokine Production by Stimulated Human Macrophages,” The Journal of Biological Chemistry, vol. 289, No. 36, pp. … [cited by applicant]
Borke et al., “Optimized triazine-mediated amidation for efficient and controlled functionalization of hyaluronic acid”, Carbohydrate Polymers 115, pp. 42-50, 2015. [cited by applicant]
Canova-Davis, Eleanor, et al., “Chemical heterogeneity as a result of hydroxylamine cleavage of a fusion protein of human insulin-like growth factor I”, Biochem. J., vol. 285, pp. 207-213, 1992. [cited by applicant]
Crimmins et al., “Chemical Cleavage of Proteins in Solutions”, Current Protocols in Protein Science, 2005 pp. 11.4.1-11.4.11 (11 pages). [cited by applicant]
D'Este et al., “A systematic analysis of DMTMM vs EDC/NHS for ligation of amines to Hyaluronan in water”, Carbohydrate Polymers 108, pp. 239-246, 2014. [cited by applicant]
E Canova-Davis et al: “Chemical heterogeneity as a result of hydroxylamine cleavage of a fusion protein of human insulin-like growth factor I”, Biochemical Journal, vol. 285, No. 1, Jul. 1, 1992 (Jul. 1, 1992), pp. 207-… [cited by applicant]
Gi Qisheng et al., “The Production and Clinical Application of Sodium Hyaluronate” Shanghai Science and Technology Press, 2012 (6 pages). [cited by applicant]
Gómez-Reyes, et al., “Metal-catalyzed hydroxylaminolysis of unactivated amide and peptide bonds,” Org. Biomol. Chem., vol. 1, 2003, pp. 866-872. [cited by applicant]
Gupta et al., “Hydrogels for wound healing applications”, Biomedical Hydrogels Biochemistry, 2011, pp. 184-227. [cited by applicant]
Hoffman, Organic Chemistry, An Intermediate Text, Second edition, 2004, p. 187 (4 pages). [cited by applicant]
International Search Report (PCT/ISA/210) issued on Jul. 31, 2017, by the European Patent Office as the International Searching Authority for International Application No. PCT/EP2017/063029 (5 pages). [cited by applicant]
International Search Report (PCT/ISA/210) mailed on Feb. 20, 2017, by the European Patent Office as the International Searching Authority for International Application No. PCT/EP2016/082778 (4 pages). [cited by applicant]
International Search Report (PCT/ISA/210) mailed on Feb. 20, 2017, by the European Patent Office as the International Searching Authority for International Application No. PCT/EP2016/082781 (4 pages). [cited by applicant]
International Search Report (PCT/ISA/210) mailed on Feb. 22, 2017, by the European Patent Office as the International Searching Authority for International Application No. PCT/EP2016/082783 (3 pages). [cited by applicant]
International Search Report (PCT/ISA/210) mailed on Feb. 23, 2017, by the European Patent Office as the International Searching Authority for International Application No. PCT/EP2016/082774 (5 pages). [cited by applicant]
International Search Report (PCT/ISA/210) mailed on May 10, 2017, by the European Patent Office as the International Searching Authority for International Application No. PCT/EP2016/082770 (5 pages). [cited by applicant]
Kurita, K., et al., “Synthetic Carbohydrate Polymers Containing Trehalose Residues in the Main Chain: Preparation and Characteristic Properties,” Macromolecules, vol. 27, 1994, pp. 7544-7549. [cited by applicant]
Lauder R.M., “Chondroitin sulphate: A complex molecule with potential impacts on a wide range of biological systems”, Complementary Therapies in Medicine, 2009, vol. 17, pp. 56-62 (7 pages). [cited by applicant]
Maleki et al., “Characterization of the chemical degradation of hyaluronic acid during chemical gelation in the presence of different cross-linker agents”, Carbohydrate Research, vol. 342, 2007 pp. 2776-2792. [cited by applicant]
Maleki et al.; “Characterization of the chemical degradation of hyaluronic acid during chemical gelation in the presence of different cross-linker agents”; Carbohydrate Research, vol. 342, pp. 2776-2792; (2007). [cited by applicant]
Olson, et al., “Specificity of Fatty Acid Acylation of Cellular Proteins,” The Journal of Biological Chemistry, vol. 260, No. 6, pp. 3784-3790 (1985). (Year: 1985). [cited by applicant]
Paterson et al., “Carbohydrate-Based Crosslinking Agents: Potential Use in Hydrogels”, Journal of Polymer Science Part A: Polymer Chemistry, vol. 49, (2011), pp. 4312-4315 (4 pages). [cited by applicant]
Sara Rydergren; “Chemical Modifications of Hyaluronan using DMTMM-Activated Amidation”; Uppsala Universitet; Aug. 2013. (English Abstract only) (35 pages). [cited by applicant]
Schante, Carole E, et al., “Chemical modification of hyaluronic acid for the synthesis of derivatives for a broad range of biomedical applications”, Carbohydrate Polymers 85, pp. 469-489, 2011. [cited by applicant]
Shimizu et al., “Cleavage of unactivated amide bonds by ammonium salt-accelerated hydrazinolysis”, ChemComm, 2014, pp. 12623-12625 (4 pages). [cited by applicant]
Shimizu et al., “Microwave-Assisted Deacylation of Unactivated Amides using Ammonium-Salt-Accelerated Transamidation”, Angewandte Communications, 2012, vol. 51, pp. 8564-8567 (4 pages). [cited by applicant]
Stern et al., “The many ways to cleave hyaluronan”, Biotechnology Advances, vol. 25, 2007, pp. 537-557 (21 pages). [cited by applicant]
Tokita et al., “Hydrolytic degradation of hyaluronic acid”, Polymer Degradation and Stability, (1995), vol. 48, pp. 269-273 (5 pages). [cited by applicant]
Tomihata, Kenji, et al., Crosslinking of hyaluronic acid with water-soluble carbodiimide Research Center for Biomedical Engineering, pp. 243-251, 1995-1996. [cited by applicant]
Tsigos et al., “Chitin deacetylases: new, versatile tools in biotechnology”, Tibtech vol. 18, pp. 305-312, Jul. 2000. [cited by applicant]
Werner et al., “Regulation of Wound Healing by Growth Factors and Cytokines”, Physiol Rev, 2003, vol. 83, pp. 835-870. [cited by applicant]
Written Opinion (PCT/ISA/210) issued on Jul. 31, 2017 by the European Patent Office as the International Searching Authority for International Application No. PCT/EP2017/063029 (6 pages). [cited by applicant]
Written Opinion (PCT/ISA/237) mailed on Feb. 20, 2017, by the European Patent Office as the International Searching Authority for International Application No. PCT/EP2016/082778 (5 pages). [cited by applicant]
Written Opinion (PCT/ISA/237) mailed on Feb. 20, 2017, by the European Patent Office as the International Searching Authority for International Application No. PCT/EP2016/082781 (6 pages). [cited by applicant]
Written Opinion (PCT/ISA/237) mailed on Feb. 22, 2017, by the European Patent Office as the International Searching Authority for International Application No. PCT/EP2016/082783 (4 pages). [cited by applicant]
Written Opinion (PCT/ISA/237) mailed on Feb. 23, 2017, by the European Patent Office as the International Searching Authority for International Application No. PCT/EP2016/082774 (7 pages). [cited by applicant]
Written Opinion (PCT/ISA/237) mailed on May 10, 2017, by the European Patent Office as the International Searching Authority for International Application No. PCT/EP2016/082770 (6 pages). [cited by applicant]
Zhu et al: “Selective cleavage of isoaspartyl peptide bonds by hydroxylamine after methyltransferase priming”, Analytical Biochemistry, Elsevier, Amsterdam, NL, vol. 364, No. 1, Mar. 31, 2007 (Mar. 31, 2007), pp. 1-7, X… [cited by applicant]
Zhu, Jeff X., et al., “Selective cleavage of isoaspartyl peptide bonds by hydroxylamine after methyltransferase priming”, Analytical Biochemistry 364, pp. 1-7, 2007. [cited by applicant]