# US 11,992,581 B2 · Methods of producing optimized gene-activated materials > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 11,992,581 B2 - Document ID: US11992581B2 - Status: Granted - Kind: B2 - Application number: 16/376,552 - Filed: 2019-04-05 - Published: 2019-07-25 - Granted: 2024-05-28 - Earliest claimed: 2016-10-07 - Assignees: Histograft, LLC - Applicants: Histograft, LLC - Inventors: Roman Vadimovich Deev; Artur Aleksandrovich Isaev; Ilya Yadigerovich Bozo; Vladimir Sergeevich Komlev - Web page: https://patentgrep.com/patent/US11992581B2 ## CPC - A61L 27/54: Biologically active materials, e.g. therapeutic substances {(A61L27/227 takes precedence)} - A61K 48/00: Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy - A61L 27/12: Phosphorus-containing materials, e.g. apatite - A61L 27/18: obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds - A61L 27/3608: {Bone, e.g. demineralised bone matrix [DBM], bone powder} - A61L 27/46: with phosphorus-containing inorganic fillers - C12Q 1/6834: Enzymatic or biochemical coupling of nucleic acids to a solid phase - A61L 2300/258: Genetic materials, DNA, RNA, genes, vectors, e.g. plasmids - A61L 2300/414: Growth factors - A61L 2430/02: for reconstruction of bones; weight-bearing implants - A61L: METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES (preservation of bodies or disinfecting characterised by the agents employed A01N; preserving, e.g. sterilising, food or foodstuffs A23; preparations for medical, dental or toiletry purposes A61K) - 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) - C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES ## Abstract Provided are gene-activated materials comprising a scaffold and at least one nucleic acid molecule which may be chemical bound together or in which the at least one nucleic acid is not bound but coated on a surface of the scaffold. Methods for regenerating bone using these gene-activated materials are also provided. ## Claims ### Claim 1 A gene-activated material comprising (i) a solid scaffold comprising a first portion of a predetermined dose of at least one nucleic acid; and (ii) a surface of the solid scaffold comprising a second portion of the predetermined dose of the at least one nucleic acid physically bound to said surface; wherein the first and second portion together comprise the predetermined dose; wherein the solid scaffold incorporating the at least one nucleic acid is produced by admixing in solution the at least one nucleic acid with scaffolding materials to produce a primary complex containing nucleic acids that are chemically-bound to the scaffolding materials and that are not removed by washing in a 0.9% solution of NaCl, and then, without washing, lyophilizing the admixture, thereby producing a gene-activated material containing a first-chemically bound fraction of the nucleic acid that is incorporated into the scaffold, which is not removed by washing in a 0.9% solution of NaCl, and a second unbound fraction of the nucleic acid that can be removed from the solid scaffold by washing in a 0.9% solution of NaCl; wherein the gene-activated material is produced by admixing the predetermined dose of the at least one nucleic acid with scaffolding materials in the presence of beta tri-calcium phosphate to produce a primary complex containing the first portion of the dose of the at least one nucleic acid; and lyophilizing or drying the solution used to prepare the scaffold, thereby producing said gene-activated material; wherein the scaffolding materials comprise beta tri-calcium phosphate and a complexing agent; and wherein the at least one nucleic acid is DNA. ### Claim 2 (depends on 1) The gene-activated material of claim 1, wherein the predetermined dose ranges from 100 to 500 μg per gram of scaffold. ### Claim 3 (depends on 1) E gene-activated material of claim 1, wherein the at least one nucleic acid is a plasmid. ### Claim 4 (depends on 1) The gene activated material of claim 1, wherein the solid scaffold further comprises at least one of Ba2+, Mg2+, and Sr2+ as the complexing agent. ### Claim 5 (depends on 1) The gene-activated material of claim 1, wherein the predetermined dose of the at least one nucleic acid is admixed with materials that form the solid scaffold prior to formation of the solid scaffold. ### Claim 6 (depends on 1) The gene-activated material of claim 1, wherein the predetermined dose of the at least one nucleic acid is admixed with the solid scaffold after synthesis of the solid scaffold. ### Claim 7 (depends on 1) The gene activated material of claim 1, wherein the first portion of the predetermined dose of at least one nucleic acid is further hound to the scaffold via complex formation with at least one of Ba2+, Mg2+, and Sr2+; and wherein the second portion of the predetermined dose of at least one nucleic acid is bound to the surface of the scaffold by drying. ### Claim 8 (depends on 7) The gene-activated material of claim 7, wherein the first portion of the predetermined dose of the at least one nucleic acid is complexed with Ca2+ and further complexed with at least one of Ba2+, Mg2+, and Sr2+; and wherein said gene-activated material provides an earlier and higher release of the predetermined dose than an otherwise identical gene-activated material not comprising Ba2+, Mg2+, and Sr2+. ### Claim 9 (depends on 1) The gene-activated material of claim 1, wherein the second portion of the at least one nucleic acid is physically bound to the surface of the scaffold in combination with at least one of glucose, dextrose, sodium hydrogen phosphate dodecahydrate, or sodium dihydrogenphosphate dehydrate. ### Claim 10 (depends on 1) The gene-activated material of claim 1 that is sterile. ### Claim 11 (depends on 1) The gene-activated material of claim 1 that is produced by admixing the predetermined dose of the at least one nucleic acid with a solid scaffold that further contains at least one complexing agent selected from the group consisting of Ba2+, Mg2+, and Sr2+, to produce a primary complex containing the first portion of the dose of the at least one nucleic acid; and lyophilizing the solution used to prepare the scaffold, thereby producing said gene-activated material.