# US 11,077,173 B2 · Lipid-based nanoparticles and methods using same > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 11,077,173 B2 - Document ID: US11077173B2 - Status: Granted - Kind: B2 - Application number: 16/916,936 - Filed: 2020-06-30 - Published: 2020-10-22 - Granted: 2021-08-03 - Earliest claimed: 2017-03-13 - Assignees: SDG, Inc. - Applicants: SDG, Inc. - Inventors: W. Blair Geho - Web page: https://patentgrep.com/patent/US11077173B2 ## CPC - A61K 38/28: Insulins - A61K 9/0019: {Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner (non-active ingredients are additionally classified in A61K47/00)} - A61K 9/127: Synthetic bilayered vehicles, e.g. liposomes or liposomes with cholesterol as the only non-phosphatidyl surfactant - A61K 47/22: Heterocyclic compounds, e.g. ascorbic acid, tocopherol or pyrrolidones - A61K 47/24: containing atoms other than carbon, hydrogen, oxygen, halogen, nitrogen or sulfur, e.g. cyclomethicone or phospholipids - A61K 47/28: Steroids, e.g. cholesterol, bile acids or glycyrrhetinic acid - Y10S 977/773: Nanoparticle, i.e. structure having three dimensions of 100 nm or less - Y10S 977/906: Drug delivery - 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) - Y10S: TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS ## Abstract The invention provides an improved lipid-based nanoparticle, which can be used to deliver a therapeutic agent to a subject, such as but not limited to a mammal, such as but not limited to a human. In certain embodiments, the nanoparticle of the invention has reduced aggregation properties as compared to those taught in the prior art. ## Claims ### Claim 1 A composition comprising a lipid-based nanoparticle, wherein the nanoparticle is enclosed by a bipolar lipid membrane, which comprises cholesterol, dicetyl phosphate, 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), stearoyl lysophosphatidylcholine, and 2,3-diacetoxypropyl 2-(5-((3aS,6aR)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamido) ethyl phosphate (biotin DHPE); wherein the membrane comprises cholesterol, dicetyl phosphate, DSPC, stearoyl lysophosphatidylcholine, and biotin DHPE in a % (w/w) ratio selected from the group consisting of: (a) about 9.4:18.1:56.8:14.1:1.5, and (b) about 8.4:16.2:47.5:7.6:1.3; wherein the biotin-DHPE extends outward from the nanoparticle; and wherein the size of the nanoparticle ranges from about 10 nm to about 150 nm. ### Claim 2 (depends on 1) The composition of claim 1, wherein the composition further comprises a therapeutic agent which is dispersed within the nanoparticle. ### Claim 3 (depends on 2) The composition of claim 2, wherein the therapeutic agent is covalently bound to the nanoparticle or wherein the therapeutic agent is not covalently bound to the nanoparticle. ### Claim 4 (depends on 2) The composition of claim 2, wherein the therapeutic agent comprises at least one selected from the group consisting of insulin, insulin analogs, interferon, parathyroid hormone, calcitonin, serotonin, serotonin agonist, serotonin reuptake inhibitor, human growth hormone, gastric inhibitory polypeptide (GIP), anti-GIP monoclonal antibody, metformin, bromocriptine, dopamine, glucagon, amylin and glucagon-like peptide-1 (GLP-1). ### Claim 5 (depends on 2) The composition of claim 2, wherein the nanoparticle is suspended in an aqueous solution comprising a free dissolved therapeutic agent that is not dispersed within the nanoparticle. ### Claim 6 (depends on 2) The composition of claim 2, wherein the therapeutic agent is insulin. ### Claim 7 (depends on 6) The composition of claim 6, wherein the nanoparticle-dispersed insulin and the free dissolved insulin are independently selected from the group consisting of insulin lispro, insulin aspart, regular insulin, insulin glargine, insulin zinc, extended human insulin zinc suspension, isophane insulin, human buffered regular insulin, insulin glulisine, recombinant human regular insulin, recombinant human insulin isophane, and any combinations thereof. ### Claim 8 (depends on 2) The composition of claim 2, further comprising at least one of the following: cellulose acetate phthalate, which is at least partially bound to the therapeutic agent dispersed within the nanoparticle; at least one charged organic molecule associated with the therapeutic agent dispersed within the nanoparticle, wherein the charged organic molecule is at least one selected from the group consisting of protamines, polylysine, a polymer with the amino acids Arg, Pro, and Thr in a mole ratio of 1:1:1, a polymer with the amino acids DL-Ala and L-Lys in a mole ratio of 6:1, histones, sugar polymers comprising a primary amino group, polynucleotides with primary amino groups, proteins comprising amino acid residues with carboxyl (COO−) or sulfhydral (S−) functional groups, and acidic polymers. ### Claim 9 A composition comprising a lipid-based nanoparticle, wherein the nanoparticle is enclosed by a bipolar lipid membrane comprising cholesterol, dicetyl phosphate, an amphipathic lipid, stearoyl lysophosphatidylcholine, and a hepatocyte receptor binding molecule; wherein the membrane comprises the cholesterol, dicetyl phosphate, amphipathic lipid, stearoyl lysophosphatidylcholine, and hepatocyte receptor binding molecule in a % (w/w) ratio such that: the cholesterol ranges from about 5% to about 15% (w/w), the dicetyl phosphate ranges from about 10% to about 25% (w/w), the hepatocyte receptor binding molecule ranges from about 0.5% to about 4% (w/w), and the stearoyl lysophosphatidylcholine ranges from about 1% to about 30% (w/w); wherein the amphipathic lipid comprises at least one selected from the group consisting of 1,2-distearoyl-sn-glycero-3-phosphocholine, 1,2-dipalmitoyl-sn-glycerol-[3-phospho-rac-(1-glycerol)], 1,2-distearoyl-sn-glycero-3-phosphoethanolamine, 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-(succinyl), 1,2-dimyristoyl-sn-glycero-3-phosphate, 1,2-dimyristoyl-sn-glycero-3-phosphocholine, 1,2-distearoyl-sn-glycero-3-phosphate, 1,2-dipalmitoyl-sn-glycero-3-phosphate, and 1,2-dipalmitoyl-sn-glycero-3-phosphocholine; wherein the at least one hepatocyte receptor binding molecule extends outward from the nanoparticle; and wherein the size of the nanoparticle ranges from about 10 nm to about 150 nm. ### Claim 10 (depends on 9) The composition of claim 9, wherein the composition further comprises a therapeutic agent which is dispersed within the nanoparticle. ### Claim 11 (depends on 10) The composition of claim 10, wherein the therapeutic agent is covalently bound to the nanoparticle or wherein the therapeutic agent is not covalently bound to the nanoparticle. ### Claim 12 (depends on 10) The composition of claim 10, wherein the therapeutic agent comprises at least one selected from the group consisting of insulin, insulin analogs, interferon, parathyroid hormone, calcitonin, serotonin, serotonin agonist, serotonin reuptake inhibitor, human growth hormone, gastric inhibitory polypeptide (GIP), anti-GIP monoclonal antibody, metformin, bromocriptine, dopamine, glucagon, amylin and glucagon-like peptide-1 (GLP-1). ### Claim 13 (depends on 10) The composition of claim 10, wherein the nanoparticle is suspended in an aqueous solution comprising a free dissolved therapeutic agent that is not dispersed within the nanoparticle. ### Claim 14 (depends on 10) The composition of claim 10, wherein the therapeutic agent is insulin. ### Claim 15 (depends on 14) The composition of claim 14, wherein the nanoparticle-dispersed insulin and the free dissolved insulin are independently selected from the group consisting of insulin lispro, insulin aspart, regular insulin, insulin glargine, insulin zinc, extended human insulin zinc suspension, isophane insulin, human buffered regular insulin, insulin glulisine, recombinant human regular insulin, recombinant human insulin isophane, and any combinations thereof. ### Claim 16 (depends on 9) The composition of claim 9, wherein the amphipathic lipid comprises at least one selected from the group consisting of 1,2-distearoyl-sn-glycero-3-phosphocholine, 1,2-dipalmitoyl-sn-glycero-3-phosphocholine, 1,2-dipalmitoyl-sn-glycero-3-[phospho-rac-(1-glycerol)], 1,2-distearoyl-sn-glycero-3-phosphoethanolamine, and 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-(succinyl). ### Claim 17 (depends on 9) The composition of claim 9, wherein the hepatocyte receptor binding molecule comprises biotin. ### Claim 18 (depends on 17) The composition of claim 17, wherein the biotin-containing hepatocyte receptor binding molecule comprises at least one selected from the group consisting of N-hydroxysuccinimide (NHS) biotin; sulfo-NHS-biotin; N-hydroxysuccinimide long chain biotin; sulfo-N-hydroxysuccinimide long chain biotin; D-biotin; biocytin; sulfo-N-hydroxysuccinimide-S-S-biotin; biotin-BMCC; biotin-HPDP; iodoacetyl-LC-biotin; biotin-hydrazide; biotin-LC-hydrazide; biocytin hydrazide; biotin cadaverine; carboxybiotin; photobiotin; ρ-aminobenzoyl biocytin trifluoroacetate; ρ-diazobenzoyl biocytin; biotin DHPE (2,3-diacetoxypropyl 2-(5-((3aS,6aR)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamido)ethyl phosphate); biotin-X-DHPE (2,3-diacetoxypropyl 2-(6-(5-((3aS,6aR)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamido)hexanamido) ethyl phosphate); 12-((biotinyl)amino)dodecanoic acid; 12-((biotinyl)amino)dodecanoic acid succinimidyl ester; S-biotinyl homocysteine; biocytin-X; biocytin x-hydrazide; biotinethylenediamine; biotin-XL; biotin-X-ethylenediamine; biotin-XX hydrazide; biotin-XX-SE; biotin-XX, SSE; biotin-X-cadaverine; α-(t-BOC)biocytin; N-(biotinyl)-N′-(iodoacetyl) ethylenediamine; DNP-X-biocytin-X-SE; biotin-X-hydrazide; norbiotinamine hydrochloride; 3-(N-maleimidylpropionyl)biocytin; ARP; biotin-1-sulfoxide; biotin methyl ester; biotin-maleimide; biotin-poly(ethyleneglycol) amine; (+) biotin 4-amidobenzoic acid sodium salt; Biotin 2-N-acetylamino-2-deoxy-β-D-glucopyranoside; Biotin-α-D-N-acetylneuraminide; Biotin-α-L-fucoside; Biotin lacto-N-bioside; Biotin—Lewis-A trisaccharide; Biotin—Lewis-Y tetrasaccharide; Biotin-a-D-mannopyranoside; and biotin 6-O-phospho-α-D-mannopyranoside. ### Claim 19 (depends on 17) The composition of claim 17, wherein the biotin-containing hepatocyte receptor binding molecule comprises at least one selected from the group consisting of biotin DHPE and biotin-X-DHPE. ### Claim 20 (depends on 10) The composition of claim 10, further comprising at least one of the following: cellulose acetate phthalate, which is at least partially bound to the therapeutic agent dispersed within the nanoparticle; at least one charged organic molecule associated with the therapeutic agent dispersed within the nanoparticle, wherein the charged organic molecule is at least one selected from the group consisting of protamines, polylysine, a polymer with the amino acids Arg, Pro, and Thr in a mole ratio of 1:1:1, a polymer with the amino acids DL-Ala and L-Lys in a mole ratio of 6:1, histones, sugar polymers comprising a primary amino group, polynucleotides with primary amino groups, proteins comprising amino acid residues with carboxyl (COO−) or sulfhydral (S−) functional groups, and acidic polymers. ### Claim 21 (depends on 9) The composition of claim 9, wherein the amphipathic lipid is 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC) and wherein the amount of the stearoyl lysophosphatidylcholine in the membrane is about 5%-30% (w/w) of the amount of DSPC in the membrane. ### Claim 22 (depends on 9) The composition of claim 9, wherein the stearoyl lysophosphatidylcholine is present in the membrane at about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, or 30% (w/w). ### Claim 23 (depends on 9) The composition of claim 9, wherein the amphipathic lipid comprises 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC). ### Claim 24 (depends on 9) The composition of claim 9, wherein the amphipathic lipid comprises 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC) and wherein the biotin-containing hepatocyte receptor binding molecule comprises at least one selected from the group consisting of biotin DHPE and biotin-X-DHPE. ### Claim 25 A method of preparing a lipid-based nanoparticle, wherein the nanoparticle is enclosed by a bipolar lipid membrane, which comprises cholesterol, dicetyl phosphate, 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), stearoyl lysophosphatidylcholine, and 2,3-diacetoxypropyl 2-(5-((3aS,6aR)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamido) ethyl phosphate (biotin DHPE); wherein the membrane comprises cholesterol, dicetyl phosphate, DSPC, stearoyl lysophosphatidylcholine, and biotin DHPE in a % (w/w) ratio selected from the group consisting of: (a) about 9.4:18.1:56.8:14.1:1.5, and (b) about 8.4:16.2:47.5:7.6:1.3; wherein the biotin-DHPE extends outward from the nanoparticle; and wherein the size of the nanoparticle ranges from about 10 nm to about 150 nm, the method comprising contacting in an aqueous system cholesterol, dicetyl phosphate, DSPC, biotin-DHPE, and stearoyl lysophosphatidylcholine. ### Claim 26 (depends on 25) The method of claim 25, wherein the nanoparticle further comprises a therapeutic agent dispersed therewithin. ### Claim 27 (depends on 26) The method of claim 26, wherein one of the following applies: (a) the therapeutic agent, cholesterol, dicetyl phosphate, DSPC, stearoyl lysophosphatidylcholine, and biotin-DHPE are simultaneously contacted in the aqueous system; (b) the nanoparticle is formed in the absence of the therapeutic agent, wherein optionally the nanoparticle is at least partially concentrated, purified or isolated, and the therapeutic agent is subsequently contacted with the nanoparticle, whereby at least a portion of the therapeutic agent is dispersed within the nanoparticle. ### Claim 28 A method of preparing a lipid-based nanoparticle, wherein the nanoparticle is enclosed by a bipolar lipid membrane comprising cholesterol, dicetyl phosphate, an amphipathic lipid, stearoyl lysophosphatidylcholine, and a hepatocyte receptor binding molecule; wherein the membrane comprises the cholesterol, dicetyl phosphate, amphipathic lipid, stearoyl lysophosphatidylcholine, and hepatocyte receptor binding molecule in a % (w/w) ratio such that: the cholesterol ranges from about 5% to about 15% (w/w); the dicetyl phosphate ranges from about 10% to about 25% (w/w); the hepatocyte receptor binding molecule ranges from about 0.5% to about 4% (w/w); the stearoyl lysophosphatidylcholine ranges from about 1% to about 30% (w/w); wherein the amphipathic lipid comprises at least one selected from the group consisting of 1,2-distearoyl-sn-glycero-3-phosphocholine, 1,2-dipalmitoyl-sn-glycerol-[3-phospho-rac-(1-glycerol)], 1,2-distearoyl-sn-glycero-3-phosphoethanolamine, 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-(succinyl), 1,2-dimyristoyl-sn-glycero-3-phosphate, 1,2-dimyristoyl-sn-glycero-3-phosphocholine, 1,2-distearoyl-sn-glycero-3-phosphate, 1,2-dipalmitoyl-sn-glycero-3-phosphate, and 1,2-dipalmitoyl-sn-glycero-3-phosphocholine; wherein the at least one hepatocyte receptor binding molecule extends outward from the nanoparticle; and wherein the size of the nanoparticle ranges from about 10 nm to about 150 nm, the method comprising contacting in an aqueous system cholesterol, dicetyl phosphate, amphipathic lipid, hepatocyte receptor binding molecule, and stearoyl lysophosphatidylcholine. ### Claim 29 (depends on 28) The method of claim 28, wherein the nanoparticle further comprises a therapeutic agent dispersed therewithin. ### Claim 30 (depends on 29) The method of claim 29, wherein one of the following applies: (a) the therapeutic agent, cholesterol, dicetyl phosphate, amphipathic lipid, hepatocyte receptor binding molecule, and stearoyl lysophosphatidylcholineare simultaneously contacted in the aqueous system; (b) the nanoparticle is formed in the absence of the therapeutic agent, wherein optionally the nanoparticle is at least partially concentrated, purified or isolated, and the therapeutic agent is subsequently contacted with the nanoparticle, whereby at least a portion of the therapeutic agent is dispersed within the nanoparticle.