Method of cleaning liposomas

专利摘要:
Pharmaceutical suspensions of liposomes containing a drug (e.g. doxorubicin hydrochloride, aminosidine sulphate or 5- fluorouracil) are prepared by dissolution of lipidic components in a solvent, addition of an aqueous solution of the drug, and emulsification by insufflation of an inert gas until the solvent is completely removed by evaporation, and then the liposomes are separated from non-entrapped drug by shaking the suspension with a polymeric ion exchange resin or a polymeric absorbent resin to absorb the non- entrapped drug, then filtering off the resin. The ion exchange resin may be a strong or weak, anionic or cationic resin. The resultant liposomic suspensions may be lyophilised.
公开号:SU1367839A3
申请号:SU802869298
申请日:1980-01-17
公开日:1988-01-15
发明作者:Моро Луиджи；Нери Гвидо；Ригамонти Алессандро
申请人:Фармиталия Карло Эрба С.П.А. (Фирма)；
IPC主号:

专利说明:

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The invention relates to medicine, in particular to methods for the preparation and purification of pharmaceutical formulations (lyophilic liposomes).
The aim of the invention is to increase the stability of the liposome by purifying the liposomal suspension using ion exchange resins.
The method is carried out as follows.
An ion exchange resin, for example, from the group of styrene, divinylbenzene, acrylic and methacrylic acid is introduced directly into a flask with a liposomal suspension, after which the contents of the flask are mixed for 10-60 minutes. Then it is filtered through a glass filter that is capable of holding an ion exchange resin, on which the unsecured preparation was adsorbed. We obtain a pure liposomal suspension, which is then lyophilized. The proposed purification method allows to obtain highly concentrated liposomal suspensions (up to 5 mg / ml of hydrochloric doxorubicin), which cannot be obtained by means of molecular sieve column chromatography (maximum 0.3 mg / ml). Moreover, this liposomal suspension is More stable and not subject to sedimentation, unlike suspensions produced by ultracentricity.
 and measures 1. In the flask for saponification, the following amount of lipids is dissolved in chloroform: 1.5 g of egg lecithin, 0.4 g of cholesterol and 0.2 of diethylphosphate, and evaporated under vacuum conditions to a dry state, hydrochloric doxorubicin solution (with concentration of 10 mg / ml) and 0.007 N buffer phosphate solution is poured into the flask, after which the resulting suspension is subjected to ultrasonic stirring for 1 min. The suspension is settled for 30 minutes under a layer of nitrogen at room temperature. Then, 2 g of resin previously activated in formic sodium and obtained by polymerization of methyl methacrylate, which forms a cross-link with divinylbenzene, which has the functionality of a carboxylic acid and has a macroporous structure, are added to it, which also allows its use in hydrophobic solutions that have a brand IRC 50 (dry weight equivalent to 5 ml of resin filled). The contents of the flask were mixed for 30 minutes, after which the suspension was filtered through a porous sheet, size G 1. Liposomes, the size of which varies from 0.5 to 2 microns and containing about 60% of the initial amount of doxorubicin, are stabilized by lyophilization.
EXAMPLE 2 The same amounts of lipids and the same dissolution conditions are used as in Example 1. Then a solution of doxorubicin is added to the flask and the resulting suspension is stirred for 10 minutes to obtain liposomes with a size of less than 1 micron. . Since the size of the liposomes was not uniform, a non-granular resin was used, which also served as a sieve. Therefore, add 10 ml of resin with the Dowex-50-X4 trademark (100-200 mesh), pre-activated in sodium formic acid. After filtration, a suspension was obtained containing liposomes with a size of 0.2-0.8 microns and comprising 75% of the initial doxorubition. Liposomes are stabilized by lyophilization. . ,
Example 3. A solution of 5-fluorouracil with a concentration of 10 mg / ml in 0.007 N phosphate buffer solution at pH 8 is poured into a saponification flask, which also contains a lipid phase, prepared analogously to example 1. The suspension obtained is treated as in example 1 using 10 ml of filter resin having the trade name Amberlite IRA 400 (C1), previously activated in chlorohydrate. The resulting liposomes were stabilized by lyophilization.
EXAMPLE 4 Liposomes are prepared analogously to Example 1, using the following lipids: 1.5 g egg lecithin, 0.4 g cholesterol, and 0.2 g stearylamine. 10 ml of Dowex-1 resin (50-100 mesh), which was previously activated, were used for purification. The resulting liposomes were stabilized by lyophilization.
PRI me R 5. All operations were carried out analogously to example 1, except that 15 g of Dowex-50YX resin (100-200 mesh) were used for cleaning, while the stirring time was increased to 40 minutes. After
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Filtering through a filter from a sintered glass G 1 was obtained a suspension of liposomes containing about 50% of the initial amount of doxorubicin. Liposomes were stabilized by lyophilization.
Example 6. All operations were carried out similarly to Example 1, except that IRC 50 (equivalent to 5 g of dry resin) was used for cleaning, stirring was carried out for 1 hour, after which the liposomal suspension was filtered on a sintered glass filter, followed by lyophilization of the product.
The proposed method is less expensive (by saving time and material) as compared with the known. In addition, it is suitable for industrial use. The resulting product is more concentrated (in a ratio of not less than 1:20 to the product obtained using gel filtration), the product is obtained 20 times more diluted when using gel filtration,



more stable, which is very important for industrial use and due to the fact that dialysis used in a known method requires at least 24 hours. In this method, the process takes about 30 minutes. Since this product remains in solution less time, it is more stable.
Data on the stability and other characteristics of liposomes, as well as processing conditions are given in the table.
15Fore Mule Invention
A method of cleaning liposomes, which includes treatment of the preparations followed by lyophilization, characterized in that, in order to increase the stability of the target product, the treatment is carried out by shaking for 10-60 minutes with an ion-exchange resin: Amberlit IRC 50 or Dowex-50, or Amberlit IRA 400 (C1), or Dowex-1 or Dauksx-50XX, and then filtered through glass.

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权利要求:
Claims (1)
[1]
Claim
The method of purification of liposomes, including the processing of drugs with subsequent lyophilization, characterized in that, in order to increase the stability of the target product, the treatment is carried out by shaking for 10-60 minutes with an ion exchange resin: Amberlite IRC = 50 or Dowex-50, or Amberlite IRA = 400 (C1), or Dowex-1, or Dowex-50, and then filtered through a glass filter. 'ι
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引用文献:

---

公开号 | 申请日 | 公开日 | 申请人 | 专利标题

---

---

DE2249552A1|1971-10-12|1973-05-30|Inchema S A|PROCESS FOR THE INCAPSULATION OF IN PARTICULAR WATER-SOLUBLE COMPOUNDS|

---

JPS5126213A|1974-08-21|1976-03-04|Tanabe Seiyaku Co| Johoseibiryushiseizaino seiho |

---

US4131815A|1977-02-23|1978-12-26|Oceanography International Corporation|Solid piezoelectric sand detection probes|

---

GB1575343A|1977-05-10|1980-09-17|Ici Ltd|Method for preparing liposome compositions containing biologically active compounds|

---

CA1116518A|1977-09-30|1982-01-19|Guido Neri|Method for purifying a liposomicsuspension|HU184141B|1979-12-27|1984-07-30|Human Oltoanyagtermelo|Adjuvant particles compositions containing said particles and biologically active substances adsorbed thereon and a process for the preparation thereof|

---

FR2521565B1|1982-02-17|1985-07-05|Dior Sa Parfums Christian|PULVERULENT MIXTURE OF LIPID COMPONENTS AND HYDROPHOBIC CONSTITUENTS, METHOD FOR PREPARING SAME, HYDRATED LIPID LAMELLAR PHASES AND MANUFACTURING METHOD, PHARMACEUTICAL OR COSMETIC COMPOSITIONS COMPRISING HYDRATED LAMID PHASES|

---

FR2553002B1|1983-10-06|1992-03-27|Centre Nat Rech Scient|IMPROVED PROCESS FOR OBTAINING UNILAMELLAR LIPOSOMES OF HIGH DIAMETERS, THEIR PHARMACOLOGICAL APPLICATION FOR THE ENCAPSULATING OF AN ACTIVE INGREDIENT FOR ITS EXTEMPORANEOUS ADMINISTRATION AND CORRESPONDING DEVICE|

---

US5736155A|1984-08-08|1998-04-07|The Liposome Company, Inc.|Encapsulation of antineoplastic agents in liposomes|

---

CA1270198A|1984-08-08|1990-06-12|Marcel B. Bally|Encapsulation of antineoplastic agents in liposomes|

---

US4755388A|1984-11-09|1988-07-05|The Regents Of The University Of California|Liposome-encapsulated 5-fluoropyrimidines and methods for their use|

---

IL79114A|1985-08-07|1990-09-17|Allergan Pharma|Method and composition for making liposomes|

---

US6406713B1|1987-03-05|2002-06-18|The Liposome Company, Inc.|Methods of preparing low-toxicity drug-lipid complexes|

---

DE69120089T2|1990-07-31|1996-12-12|Liposome Co Inc|Accumulation of amino acids and peptides in liposomes|

---

DE4107152C2|1991-03-06|1994-03-24|Gregor Cevc|Preparations for non-invasive administration of antidiabetics|

---

DE4107153A1|1991-03-06|1992-09-10|Gregor Cevc|Compsns. for application of active agents|

---

JPH04127874U|1991-05-13|1992-11-20|

---

JPH04134673U|1991-06-07|1992-12-15|

---

JPH09502700A|1993-04-02|1997-03-18|ザリポソームカンパニー、インコーポレーテッド|Method for producing liposome|

---

IT1270678B|1994-10-20|1997-05-07|Bayer Ag|KETOPROFEN LIPOSOMES|

---

DE19639811A1|1996-09-27|1998-04-02|Artur Herzog Dr Mesmer|Use of a liposome solution to enhance the effectiveness and / or decrease the toxicity of drugs|

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法律状态:

优先权:

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申请号 | 申请日 | 专利标题

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IT19434/79A|IT1110989B|1979-01-19|1979-01-19|PHARMACEUTICAL FORMS CONSTITUTED BY LIPOSOMES AND RELATED PROCEDURES|

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