Liposome of irinotecan or its hydrochloride and preparation method thereof

Abstract

A liposome of irinotecan or irinotecan hydrochloride and its preparation method are disclosed. The liposome contains irinotecan or irinotecan hydrochloride, neutral phospholipid and cholesterol, wherein the weight ratio of the cholesterol to the neutral phospholipid is 1:3 to 1:5. The liposome is prepared by an ion gradient method.

1 .- 23 . (canceled) 
     
     
         24 . A liposome of irinotecan or irinotecan hydrochloride, comprising irinotecan or irinotecan hydrochloride, a neutral phospholipid and cholesterol, wherein the weight ratio of the cholesterol to the neutral phospholipid is 1:3 to 1:5. 
     
     
         25 . The liposome according to  claim 24 , wherein the weight ratio of the neutral phospholipid to the irinotecan or the irinotecan hydrochloride is 2:1 to 5:1. 
     
     
         26 . The liposome according to  claim 24 , wherein the neutral phospholipid comprises hydrogenated soybean phosphatidylcholine. 
     
     
         27 . The liposome according to  claim 26 , wherein the neutral phospholipid consists of the hydrogenated soybean phosphatidylcholine. 
     
     
         28 . The liposome according to  claim 24 , wherein the weight ratio of the cholesterol to the neutral phospholipid is 1:3.5 to 1:4.5. 
     
     
         29 . The liposome according to  claim 24 , wherein the liposome is prepared by an ion gradient method. 
     
     
         30 . The liposome according to  claim 29 , wherein the liposome has an ion gradient formed between the internal water phase and the external water phase of the liposome. 
     
     
         31 . The liposome according to  claim 24 , wherein the liposome further comprises a lipid derivative of a hydrophilic polymer. 
     
     
         32 . The liposome according to  claim 31 , wherein the weight ratio of the lipid derivative of the hydrophilic polymer to the irinotecan or the irinotecan hydrochloride is 0.2 to 0.4. 
     
     
         33 . The liposome according to  claim 24 , wherein the liposome further comprises a charged phospholipid selected from the group consisting of dilauroyl phosphatidylglycerol, dipalmitoyl phosphatidylglycerol, distearoyl phosphatidylglycerol, dimyristoyl phosphatidylglycerol, dioleic acid phosphatidylserine, dioleoyl phosphatidylglycerol, dilauroyl phosphatidic acid, dimyristoyl phosphatidic acid, distearoyl phosphatidic acid and a mixture thereof, and the weight ratio of the charged phospholipid to the neutral phospholipid is 1:5 to 1:100. 
     
     
         34 . The liposome according to  claim 24 , comprising: 
       
         
           
                 
                 
                 
               
                     
                 
                   irinotecan hydrochloride 
                   1 
                   part by weight; 
                 
                   hydrogenated soybean 
                   3.4-3.8 
                   parts by weight; 
                 
                   phosphatidylcholine 
                 
                   polyethylene glycol 2000-distearoyl 
                   0.34-0.38 
                   part by weight; and 
                 
                   phosphatidyl ethanolamine 
                 
                   cholesterol 
                   0.8-0.95 
                   part by weight, 
                 
                     
                 
             
                
               
               
                
                
                
                
                
                
                
               
            
           
         
         wherein the weight ratio of the cholesterol to the hydrogenated soybean phosphatidylcholine is 1:4. 
       
     
     
         35 . A method of preparing a liposome of irinotecan or irinotecan hydrochloride, comprising:
 1) preparing a blank liposome comprising a neutral phospholipid and cholesterol, wherein the weight ratio of the cholesterol to the neutral phospholipid is 1:3 to 1:5;   2) replacing the external water phase of the blank liposome to obtain a blank liposome having an ionic gradient between the internal water phase and the external water phase of the blank liposome;   3) preparing an aqueous solution of the irinotecan or the irinotecan hydrochloride;   4) mixing the aqueous solution with the blank liposome having the ionic gradient in a dispersion; and   5) incubating the dispersion with heating and stirring to obtain a preparation comprising the liposome of irinotecan or irinotecan hydrochloride.   
     
     
         36 . The method according to  claim 35 , further comprising:
 6) removing non-encapsulated irinotecan or irinotecan hydrochloride from the preparation; and   7) concentrating the non-encapsulated irinotecan or irinotecan hydrochloride removed preparation.   
     
     
         37 . The method according to  claim 35 , wherein the blank liposome has a desired particle size and is prepared using a method selected from the group consisting of methods A, B, C and D:
 A. dissolving the neutral phospholipid and the cholesterol in anhydrous ethanol or a mixed solvent of anhydrous ethanol and tert-butyl alcohol to obtain a solution; mixing the solution with a buffer to obtain a mixture; obtaining a crude blank liposome after removing the ethanol from the mixture through a reduced pressure distillation; and obtaining the blank liposome from the crude blank liposome using at least one of a high-pressure homogenizer and an extrusion equipment;   B. dissolving the neutral phospholipid and the cholesterol in chloroform or a mixed solvent of chloroform and methanol to obtain a solution; forming a lipid film from the solution using rotary evaporation; obtaining a crude blank liposome after hydrating the lipid film with a buffer; and preparing the blank liposome from the crude blank liposome using at least one of a high-pressure homogenizer and an extrusion equipment;   C. mixing the neutral phospholipid, the cholesterol and a buffer to obtain a mixture; preparing the blank liposome from the mixture using at least one of a high-pressure homogenizer and an extrusion equipment; and   D. dissolving the neutral phospholipid and the cholesterol in anhydrous ethanol or a mixed solvent of anhydrous ethanol and tert-butyl alcohol to obtain a solution; mixing the solution with a buffer to obtain a mixture; and preparing the blank liposome from the mixture using at least one of a high-pressure homogenizer and an extrusion equipment;   wherein the buffer is selected from the group consisting of a buffer comprising Na + , K + , Fe 2+ , Ca 2+ , Ba 2+ , Mg 2+ , Li + , NH 4 + , H 4+  ion salts and a mixture thereof.   
     
     
         38 . A liposome injection comprising the liposome of irinotecan or irinotecan hydrochloride according to  claim 24 . 
     
     
         39 . The liposome injection according to  claim 38 , wherein the injection comprises a stabilizer selected from the group consisting of ethylene diamine tetraacetic acid, ethylene diamine tetraacetic acid disodium, ethylene diamine tetraacetic acid dicalcium and a mixture thereof; and the amount of the stabilizer is more than 0% (w/v), but less than or equal to 0.5% (w/v), of the liposome injection. 
     
     
         40 . The liposome injection according to  claim 38 , wherein the injection is a liquid injection or a lyophilized powder for injection. 
     
     
         41 . The liposome injection according to  claim 38 , wherein the injection comprises an osmotic pressure regulator selected from the group consisting of glucose, sucrose, sorbitol, mannitol, sodium chloride, glycerine, histidine, histidine hydrochloride, glycine, glycine hydrochloride, lysine, serine, glutamic acid, arginine, valine and a mixture thereof; and the amount of the osmotic pressure regulator is more than 0% (w/v), but less than or equal to 5% (w/v), of the liposome injection. 
     
     
         42 . The liposome injection according to  claim 38 , wherein the injection further comprises an antioxidant selected from the group consisting of a water-soluble antioxidant and an oil-soluble antioxidant; wherein the oil-soluble antioxidant is selected from the group consisting of α-tocopherol, α-tocopherol succinate, α-tocopherol acetate and a mixture thereof; the water-soluble antioxidant is selected from the group consisting of ascorbic acid, sodium bisulfite, sodium sulfite, sodium pyrosulfite, L-cysteine and a mixture thereof; and the amount of the antioxidant is more than 0% (w/v), but less than or equal to 0.5% (w/v), of the liposome injection. 
     
     
         43 . The liposome injection according to  claim 40 , wherein the injection is the lyophilized powder for injection comprising a lyoprotectant, and the lyophilized powder is prepared by freeze-drying. 
     
     
         44 . The liposome injection according to  claim 38 , comprising: 
       
         
           
                 
                 
                 
               
                     
                 
                   irinotecan hydrochloride 
                   1 
                   part by weight; 
                 
                   hydrogenated soybean 
                   3.4-3.8 
                   parts by weight; 
                 
                   phosphatidylcholine 
                 
                   polyethylene glycol 2000-distearoyl 
                   0.34-0.38 
                   part by weight; 
                 
                   phosphatidyl ethanolamine 
                 
                   cholesterol 
                   0.8-0.95 
                   part by weight; and 
                 
                   ethylene diamine tetraacetic acid 
                   0.05-0.09 
                   part by weight 
                 
                   disodium 
                 
                     
                 
             
                
               
               
                
                
                
                
                
                
                
                
                
               
            
           
         
         wherein the weight ratio of the cholesterol to the hydrogenated soybean phosphatidylcholine is 1:4. 
       
     
     
         45 . A method of preparing a liposome injection comprising a liposome of irinotecan or irinotecan hydrochloride, the method comprising preparing the liposome using the method of  claim 35 . 
     
     
         46 . The method according to  claim 45 , further comprising:
 adjusting the concentration of the irinotecan or the irinotecan hydrochloride in the injection, metering the volume of the injection, sterilizing the injection by filtration, filling the sterilized injection to vials and sealing the vials to obtain a liquid injection; or   adding a lyoprotectant to the injection, adjusting the concentration of the irinotecan or the irinotecan hydrochloride in the injection, metering the volume of the injection, sterilizing the injection by filtration, filling the sterilized injection to vials, sealing the vials, freeze-drying the sealed vials to obtain a lyophilized power for injection.   
     
     
         47 . The liposome according to  claim 24 , wherein the weight ratio of the neutral phospholipid to the irinotecan or the irinotecan hydrochloride is 2.5:1 to 4:1. 
     
     
         48 . The liposome according to  claim 24 , wherein the weight ratio of the cholesterol to the neutral phospholipid is 1:4. 
     
     
         49 . The liposome according to  claim 30 , wherein the internal water phase of the liposome has a higher ion concentration than the external water phase of the liposome. 
     
     
         50 . The liposome according to  claim 31 , wherein the hydrophilic polymer is DSPE-PEG 2000 . 
     
     
         51 . The liposome injection according to  claim 39 , wherein the stabilizer is ethylene diamine tetraacetic acid disodium.