# US 10,500,165 B2 · Purified therapeutic nanoparticles and preparation methods thereof > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 10,500,165 B2 - Document ID: US10500165B2 - Status: Granted - Kind: B2 - Application number: 14/790,646 - Filed: 2015-07-02 - Published: 2016-01-07 - Granted: 2019-12-10 - Earliest claimed: 2014-07-03 - Assignees: Cspc Zhongqi Pharmaceutical Technology (Shijiazhuang) Co., Ltd. - Applicants: Cspc Zhongqi Pharmaceutical Technology (Shijiazhuang) Co., Ltd. - Inventors: Chunlei Li; Yanhui Li; Min Liang; Caixia Wang; Yajuan Wang; Shixia Wang; Dongjian Chen; Yongfeng Li - Web page: https://patentgrep.com/patent/US10500165B2 ## CPC - A61K 9/5169: {Proteins, e.g. albumin, gelatin} - A61K 31/337: having four-membered rings, e.g. taxol - A61P 35/00: Antineoplastic agents - B82Y 5/00: Nanobiotechnology or nanomedicine, e.g. protein engineering or 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) - A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS - B82Y: SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES ## Abstract Purified therapeutic nanoparticles are provided herein. Such nanoparticles comprise an active pharmaceutical ingredient and human serum albumin, wherein the weight ratio of human serum albumin to the active ingredient in the therapeutic nanoparticles is from 0.01:1 to 1:1, and wherein the nanoparticles are substantially free of free human serum albumin that is not incorporated in the nanoparticles. The present disclosure also provides pharmaceutical compositions that comprise the purified therapeutic nanoparticles and are also substantially free of free human serum albumin. Methods for preparing and using the purified therapeutic nanoparticles and compositions thereof are also provided. ## Claims ### Claim 1 Purified therapeutic nanoparticles, consisting essentially of paclitaxel and human serum albumin (HSA), wherein the weight ratio of HSA to paclitaxel in the therapeutic nanoparticles is from 0.03:1 to 0.95:1, wherein less than 10% HSA in the therapeutic nanoparticles is free HSA that is not incorporated into the nanoparticles, wherein the therapeutic nanoparticles contain less than 0.05 mg/ml organic solvent(s), and wherein when suspended in 0.9% sodium chloride or 5% mannitol, the resulting suspension is stable for 48 hours at room temperature. ### Claim 2 (depends on 1) A pharmaceutical composition, comprising the purified therapeutic nanoparticles of claim 1, wherein less than 10% of HSA in the composition is free HSA that is not incorporated in the nanoparticles. ### Claim 3 (depends on 2) The pharmaceutical composition of claim 2, wherein the weight ratio of HSA to paclitaxel of the therapeutic nanoparticles is from 0.03:1 to 0.7:1, and wherein the average particle size of the therapeutic nanoparticles is from 50 nm to 190 nm. ### Claim 4 (depends on 2) The pharmaceutical composition according to claim 2, wherein the pharmaceutical composition is in the form of liquid or lyophilized powder. ### Claim 5 (depends on 2) The pharmaceutical composition according to claim 2, wherein the pharmaceutical composition also comprises lyophilization excipient when the pharmaceutical composition is in the form of lyophilized powder. ### Claim 6 (depends on 5) The pharmaceutical composition according to claim 5, wherein the lyophilization excipient is selected from one or more of mannitol, sucrose, lactose, maltose, trehalose, and dextran. ### Claim 7 (depends on 2) The pharmaceutical composition according to claim 2, wherein at most 5% of the HSA in the composition is free HSA (by weight) that is not incorporated in the nanoparticles. ### Claim 8 (depends on 2) The pharmaceutical composition according to claim 2, wherein the weight ratio of HSA to paclitaxel of the therapeutic particles is from 0.03:1 to 0.7:1. ### Claim 9 (depends on 2) The pharmaceutical composition according to claim 2, wherein the therapeutic nanoparticles contain less than 5 ug/ml organic solvent(s). ### Claim 10 (depends on 2) The pharmaceutical composition according to claim 2, wherein less than 1% of the HSA in the composition is free HSA (by weight) that is not incorporated in the nanoparticles. ### Claim 11 (depends on 2) The pharmaceutical composition according to claim 2, wherein the average particle size of the therapeutic nanoparticles is from 100 nm to 160 nm. ### Claim 12 (depends on 2) The pharmaceutical composition of claim 2, wherein the weight ratio of HSA to paclitaxel is from 0.10:1 to 0.6:1. ### Claim 13 (depends on 2) The pharmaceutical composition according to claim 2, wherein the organic solvent(s) is a pure solvent having low water-solubility and a low boiling point or a mixture of a solvent having low water-solubility and a low boiling point with a small molecular alcohol. ### Claim 14 (depends on 2) The pharmaceutical composition according to claim 2, wherein the weight ratio of HSA to paclitaxel in therapeutic nanoparticles is from 0.04:1 to 0.75:1. ### Claim 15 (depends on 2) The pharmaceutical composition according to claim 2, wherein the average particle size of the therapeutic nanoparticles is from 30 to 200 nm. ### Claim 16 (depends on 1) A method for preparing the purified therapeutic nanoparticles of claim 1, comprising: 1) dissolving paclitaxel in organic solvent to form an oil phase, and dissolving human serum albumin in water to form an aqueous phase; 2) forming an oil-in-water emulsion using the oil phase and aqueous phase of step 1); 3) removing the organic solvent in the emulsion to obtain a suspension containing the therapeutic nanoparticles; and 4) removing free HSA that is not incorporated in the nanoparticles from the suspension to obtain purified therapeutic nanoparticles. ### Claim 17 (depends on 16) The method according to claim 16, wherein the organic solvent is selected from one or more of chloroform and ethanol. ### Claim 18 (depends on 16) The method according to claim 16, further comprising: between steps 3) and 4), a step of dialyzing the suspension of step 3) with an aqueous solution to remove remaining organic solvent from the suspension. ### Claim 19 (depends on 18) The method according to claim 18, wherein the aqueous solution is water. ### Claim 20 (depends on 16) The method according to claim 16, wherein said separating in step 4) is conducted using a method selected from: centrifugation, dialysis, and exclusion chromatography. ### Claim 21 (depends on 2) A method for preparing the pharmaceutical composition according to claim 2, comprising: 1) dissolving paclitaxel in organic solvent to form an oil phase, and dissolving human serum albumin in water to form an aqueous phase; 2) forming an oil-in-water emulsion using the oil phase and aqueous phase of step 1); 3) removing the organic solvent in the emulsion to obtain a suspension containing the therapeutic nanoparticles; 4) removing free HSA that is not incorporated in the nanoparticles to obtain purified therapeutic nanoparticles; 5) re-suspending the purified therapeutic nanoparticles in an excipient-containing solution; and 6) optionally lyophilizing the re-suspension of the purified therapeutic nanoparticles to obtain the pharmaceutical composition. ### Claim 22 (depends on 21) The method of claim 21, further comprising: between steps 3) and 4), a step of dialyzing the suspension of step 3) with an aqueous solution to remove remaining organic solvent from the suspension. ### Claim 23 (depends on 22) The method according to claim 22, wherein the aqueous solution is water. ### Claim 24 (depends on 2) A method for preparing the pharmaceutical composition according to claim 2, comprising: 1) dissolving paclitaxel in organic solvent to form an oil phase, and dissolving human serum albumin in water to form an aqueous phase; 2) forming an oil-in-water emulsion using the oil phase and aqueous phase of step 1); 3) removing the organic solvent in the emulsion to obtain a suspension containing the therapeutic nanoparticles; 4) dialyzing the suspension obtained after removal of the organic solvent by an excipient-containing solution to remove free HSA that is not incorporated in the nanoparticles; and 5) optionally lyophilizing the dialyzed suspension to obtain the pharmaceutical composition. ### Claim 25 (depends on 24) The method of claim 24, further comprising: between steps 3) and 4), a step of dialyzing the suspension of step 3) with an aqueous solution to remove remaining organic solvent from the suspension. ### Claim 26 (depends on 25) The method of claim 25, wherein the aqueous solution is water. ### Claim 27 (depends on 2) A method for treating cancer, comprising: administering a therapeutically effective amount of the pharmaceutical composition according to claim 2 to a subject in need thereof. ### Claim 28 (depends on 3) A method for treating cancer, comprising: administering a therapeutically effective amount of the pharmaceutical composition according to claim 3 to a subject in need thereof.