# US 11,306,296 B2 · MG53 mutants, methods of making the same, and uses thereof > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 11,306,296 B2 - Document ID: US11306296B2 - Status: Granted - Kind: B2 - Application number: 16/262,833 - Filed: 2019-01-30 - Published: 2019-05-23 - Granted: 2022-04-19 - Earliest claimed: 2016-08-01 - Assignees: Hope Medicine (Nanjing) Co., Ltd. - Applicants: Hope Medicine (Nanjing) Co., Ltd. - Inventors: Rui-ping Xiao; Fengxiang Lu; Yan Zhang; Sile Guo - Web page: https://patentgrep.com/patent/US11306296B2 ## CPC - C12N 9/104: {Aminoacyltransferases (2.3.2)} - A61K 38/17: from animals; from humans {(enzyme inhibitors A61K38/005)} - A61P 9/00: Drugs for disorders of the cardiovascular system - A61P 9/10: for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis - A61P 13/12: of the kidneys - C07K 14/47: from mammals - A61K 38/00: Medicinal preparations containing peptides (peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00) - C12Y 203/02: Aminoacyltransferases (2.3.2) - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D) - A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) - C12Y: ENZYMES ## Abstract The present invention relates to an MG53 mutant, wherein the MG53 mutant is identical to the amino acid sequence of a wild-type MG53 except for at least one serine in the coiled-coil-SPRY region of the wild-type MG53, which is deleted and/or mutated into any other non-serine or non-threonine amino acid(s). The present invention also relates to a pharmaceutical composition comprising the MG53 mutant, a nucleic acid encoding the MG53 mutant, a method for preparing the MG53 mutant, use of the MG53 mutant in the manufacture of a medicament for treating heart diseases, diabetic cerebrovascular diseases, diabetic ocular complications, diabetic neuropathy, diabetic foot, kidney diseases, and diseases associated with cellular and/or tissue damage. In particular, the MG53 mutant of the present invention may avoid or reduce metabolic side effects, such as, insulin resistance, obesity, diabetes, hypertension, dyslipidemia, etc., brought by the wild-type MG53, while treating heart diseases, diabetic cerebrovascular diseases, diabetic ocular complications, diabetic neuropathy, diabetic foot, kidney diseases, and diseases associated with cellular and/or tissue damage. ## Claims ### Claim 1 A Mitsugumin 53 (MG53) mutant, wherein the MG53 mutant is identical to the amino acid sequence of a wild-type MG53 except for at least one serine in the coiled-coil-SPRY region of the wild-type MG53, which is deleted and/or mutated into any other non-serine or non-threonine amino acid(s), wherein the deleted or mutated serine(s) is(are) located at one or more of the following positions of the amino acid sequence of the wild-type MG53: positions 255, 305, 306, and 307, and wherein the wild-type MG53 is a human, mouse, rat, monkey, swine, or dog wild-type MG53, the MG53 mutant has an amino acid sequence having at least 90% homology to any one of the amino acid sequences set forth in SEQ ID NOs: 7-12 or 147-150, and the MG53 mutant avoids or reduces metabolic side effects compared to the wild-type MG53, while still retaining cell repair function and/or cardioprotective function. ### Claim 2 (depends on 1) The MG53 mutant according to claim 1, wherein the coiled-coil-SPRY region is located at positions 122-477 of the amino acid sequence of the wild-type MG53. ### Claim 3 (depends on 1) The MG53 mutant according to claim 1, wherein the amino acid sequence of the wild-type MG53 is set forth in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 139, SEQ ID NO: 140, SEQ ID NO: 141, or SEQ ID NO: 142. ### Claim 4 (depends on 1) The MG53 mutant according to claim 1, wherein the serine is mutated into a non-polar amino acid. ### Claim 5 (depends on 4) The MG53 mutant according to claim 4, wherein the non-polar amino acid is selected from the group consisting of glycine, alanine, leucine, isoleucine, valine, proline, phenylalanine, methionine and tryptophan. ### Claim 6 (depends on 1) The MG53 mutant according to claim 1, wherein the serine is mutated into a non-serine or non-threonine polar amino acid. ### Claim 7 (depends on 6) The MG53 mutant according to claim 6, wherein the polar amino acid is selected from the group consisting of glutamine, cysteine, asparagine, tyrosine, aspartic acid, glutamic acid, lysine, arginine and histidine. ### Claim 8 (depends on 1) The MG53 mutant according to claim 1, wherein the deleted or mutated serine is located at position 255 of the amino acid sequence of the wild-type MG53, or the deleted or mutated serines are located at positions 305, 306 and 307 of the amino acid sequence of the wild-type MG53. ### Claim 9 (depends on 1) The MG53 mutant according to claim 1, wherein the MG53 mutant has an amino acid sequence having at least 95% homology to any one of the amino acid sequences set forth in SEQ ID NOs: 7-12 or 147-150. ### Claim 10 A MG53 mutant, wherein the amino acid sequence of the MG53 mutant is any one of the amino acid sequences set forth in SEQ ID NOs: 7-12 or 147-150. ### Claim 11 (depends on 10) The MG53 mutant according to claim 10, wherein the amino acid sequence of the MG53 mutant is the amino acid sequence set forth in SEQ ID NO: 7. ### Claim 12 (depends on 1) A pharmaceutical composition comprising the MG53 mutant of claim 1 and a pharmaceutically acceptable carrier. ### Claim 13 (depends on 1) An isolated nucleic acid comprising a nucleic acid sequence encoding the amino acid sequence of the MG53 mutant of claim 1. ### Claim 14 (depends on 13) The nucleic acid according to claim 13 comprising any one of the nucleic acid sequences set forth in SEQ ID NOs: 13-18 or 151-154. ### Claim 15 (depends on 13) An expression vector comprising the nucleic acid sequence according to claim 13. ### Claim 16 (depends on 15) A host cell comprising the expression vector according to claim 15. ### Claim 17 (depends on 1) A method for preparing the MG53 mutant according to claim 1, comprising determining one or more serine positions for mutation, performing site-directed mutagenesis at the position on the full-length sequence of a plasmid comprising the nucleic acid sequence encoding the amino acid sequence of wild-type MG53, transfecting the plasmid with site-directed mutagenesis into a host cell, and inducing the host cell to produce the MG53 mutant. ### Claim 18 (depends on 1) A method for treating heart diseases, diabetic cerebrovascular diseases, diabetic ocular complications, diabetic neuropathy, diabetic foot, kidney diseases, and diseases associated with cellular and/or tissue damage, comprising administering to a subject in need thereof a therapeutically effective amount of the MG53 mutant of claim 1. ### Claim 19 (depends on 18) The method according to claim 18, wherein the MG53 mutant avoids metabolic side effects while treating heart diseases, diabetic cerebrovascular diseases, diabetic ocular complications, diabetic neuropathy, diabetic foot, kidney diseases, and diseases associated with cellular and/or tissue damage. ### Claim 20 (depends on 18) The method according to claim 18, wherein the amino acid sequence of the MG53 mutant is the amino acid sequence set forth in SEQ ID NO: 7.