前苏联专利SU1158031A3 Method of obtaining liposoms

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专利摘要:
1. METHOD OF FLOORING OF LIPOSOM, including dispersing the encapsulated liquid in the presence of a phosphor-iOpidOB and a solution of a surfactant substance in a vololerasoluble organic solvent by means of ultrasound and removing the solvent, about 1 and-1 and 10. so that, in order to simplify the process, after dispersion, water or an aqueous salt solution is added to the resulting mixture, emulsified by stirring, and the solvent is removed by passing gas and distillation or distillation at a lower pressure before or after emulsification. 2. Method IO of clause 15 of a tl and h and y and the fact that as a water-insoluble solvent use benzene, alkyl- and gas-benzene benzene, and.piphatic zfira, a.aldehydes, esters, ketones, free and halogenated aliphatic and cycloaliphatic carbons. 3, the method according to p. 1, about tl and h and 10 u and with the fact that as lipids are used substances from the group of lecithin, phosphates, ethanolamine, lysolecithin. lysophosphate 1 vdil-ethanolamine, phosphatidyl serine, phosphatidyl inositol, sphingomyelin, cardiolipi a, phosphate acid, cerebroside, stearylamine, dipalmitoyl phosphatidyl choline, or a mixture of a single phosphodipid, cholesterol, 4 topherol, nitric nitroyl phosphatidylcholine, or a mixture of one phosphoripid, cholesterol, tocopherol, acetyl amine M SL solution of sodium chloride. 00 5. The method according to claim 1, of which is that dyes5 aromatic substances, drugs, enzymes or polypetides are used as the encapsulating substance.
公开号:SU1158031A3
申请号:SU782649649
申请日:1978-08-04
公开日:1985-05-23
发明作者:Шнейдер Мишель
申请人:Баттель Мемориал Институт (Фирма)；
IPC主号:

专利说明:

111
5grpcdrrrrrn is related to medicine, namely g, 1 liposome is obtained by way of
Aim and gain simplification of the way
Lp and measures KB 10 milliliter rr pivxcoiiyio flask interfere with 2 ml of jpJCVTH.TOTforo ether and 1 ml of u, wp.O PKc; 3Ha then add 0.2 ml of liHcyjniHOBoro solution of 10 mg / l in p. One 0% NaCC- at pH 3. Then, 150 mg of dipalmitoylphosphatidylcholine (hydrophilic lipophilic compound) is released, and at room temperature: the temperature of the homogeneous mixture noABP.prsjciT to the action of ultrasound in
t-c-cheni-NiHH with the help of the BRANSON generator (r-model B-12, 20 kHz, 150 W), according to a transparent rastg: op, its;, kl.a.a; ky liposomal preceding HHrjft-r G: g : id, e microbubble insu .niHCBi Fo solution. Such microbubbles g, /: c: j,;, iipyK) T in the colloidal G (; yyy) .- g .: - n1- in an organic solvent, o / ;, h: h / dem excess dissolved :; 5 Oierhio1: tth active substance.
. ::. Srg anic solution pod gg; ;;,RU. flask containing 30 ml:,. ;: -; o aqueous Na (C - concentration;, nc;, 9 / i Si; mixture emulsified;. ;; -: 3 I. with ii Mooinbio emulsifying that hl: -; f uui:; n, rotating with high o, or o -.; .--.; after this stage the mixture can be; :: g, 1, ep}); at dispersion of small kaiii -n / v (p3 -: ical solvent in
Calsda droplets in dispersion: liper described liposomes
apejvierl vrn ;; iki and excess lipids.
. g :: omp ds.kt trubkl flow of air-xi ICi.u.asor inutry kolb and pass: - (e, bl1-1; co f; hyperiostrum; emulsions v; .-) H- -v-pr- MeLTT-iBaHHii at 30 ° C; mixture - oz, z (yk to gtars solvent cooled-: og; - condenser at -IOC, in bVfrie of which the vapors are condensed -; (co ::. 1b, this operatdao continues -i; ri, until kkdeis .Л1, |; and (approximately 20-30).
. ;; i; - of this stage, a flask of 20 ml of clear solution (NaC f,) remains in the flask.
2 chg of the liposome solution was subjected to chromatography on a ionic bath containing 2.5 g of Sephadex G-50 (eluent: 0.15 M aqueous NaCtj Oj, 05 M phosphate buffer, pI7.5 Irm analysis (spectrometry) elute was determined that only 20 25% peroxal insulin solution
incaiculir is beaten up (r. "Therefore, in most cases of cases, the Lipscomm solution dissolves cryptic disease) by way of you, as if pi; ;-; en. BS: Og, g (cleaning.
i p and M e 15 2, Vodl-i and rstvor and; yuglmkozi. da1b 10 g / l d; 1СГ1ергчую in 0.15 M NaCt solution as follows. A mixture of 7A mg of lepitol, 0.1.L of amylog7-glucosidase of pacTnofia vi and 3 ml of diisopropyl alcohol was treated with ultrasound for 10 minutes; 2: un (20 kHz, 1 5Г1 W) rip and cooling below 30 ° C with the help of cooling: bath. 1 rush: from transparent i th o h e n i y t; ;; i u b a v a g yvi g.; 1 к о с т i X KOTOp-oii ext; and; | I; -t 15 ml of 0.15 M aqueous / vaC5. (Mons, sig |) HcIcir og as oggisano in npiiMt pc 1, ate something Topkuyu chmulsigo about ;; squeezed by a stream of nitrogen, which evaporates the organic 1-g11 solvent. This product. 1g / ka; from until then. until evaporation ceases (20-30 mime). A clear suspension of colloidal chppos in water is obtained; O, 15 M solution of l.Sy, s. Content 1e -; a very small amount (example (; 5/0 uncovered enzyme „
Depending on the purpose, this solution can be used}) by itself only after separating out ;; a:; sulirovanie, ag lgococides (Hanpiimer, by means of chromatography on Sephadex gel).
Irina p 3. The method of example 1 is repeated, but 0.05 1-1 aqueous 6y (bep: ii; i yjacTBOp (phosphate buffer 10 mmol, pH 7.2) 0.5 MF) is used as an encapsulated solution. MJi cytosis1- narab1p) PS. Dispersion of 1 day phase1 {ete / 0.15 M JACCC (10 1-1: 1), phpidom - car; i1olipin (i7 mg) 5 organic solvent - dibutylone simple 3p1: p (2.4 ml) and SCS , (0.6 ml).
Example, The method of example 1 is repeated, but using the following 1 (their substances: an encapsulated concentration of 100 mg / ml (0.1 mol) encapsulated with a penicillon of 0.15 M Huass, a surfactant - lecithin 105 mg, cholesterol 35 mg, butyl acetate solvent 3 ml.
Example 5. The method of Example 1 is repeated, but the following ingredients are used: for the encapsulation, betametal acid concentration 15P mg / l in a mixture of sodium phosphate (0.05 g-1 l); the dispersing phase is 0.15 M A / aC {; lipi / - lecithin 85 mg and phosphatidyl-etaiolamine / 45 mg; solvent 3-heptanone 3 ml.
Example 6. The method according to npiepy 1 is repeated using the following substances as ingredients: 0.1 ml of a diluted solution of a polyiscipulatory – gyuli111 dilovy KVicjioT (poly J) –poly (c) concentration of 1 mg / ml complex to encapsulate j the dispersing phase is 0.15 M / JaCP surfactant — lecithin 60 mg, stearylamine 20 mg and cholesterol 15 mg, the solvent is a 1: 1 mixture by volume of diisopropyl tfir and butyl acetate 3 ml.
Example 7. 1.5 g of lecithin, 0.4 g of phosphatid serine and 0.5 g of cholesterol 15a, then a solution of 1.8 mg of actinomycin D in 3 ml of phosphate buffer (0.1 M, pH 7); The mixture according to; 1 was subjected to ultrasonic treatment for 5 minutes, as described in Example 1, 100 ml of aqueous phosphate buffer (0.1 K, pH 7) were then added and the dilution was carried out as in the example; 1. Do not stop the emulsifier, the flask, the flask is attached to a vacuum trap and the pressure is gradually reduced to 10 Torr while maintaining the temperature at 20-22 ° C. A field of about D5 min diisoylyl ether is completely off. The residual mixture is obtained in the form of a prose (1) proprietary JiiiiiocoMHoro solution. By means of chromatography of the sample on Ce (1) adex, we find that 88% of the initial actinium is captured.
Example 8, 1 g of trypsin in 30 ml of 0.1 M (pil 7 of phosphate buffer was treated with ultrasound in the presence of a solution of 30 g of phosphatidyl inositol in 100 ml of dipropyl simple sephira. Then 460 l of 0.5% aqueous L / aC and wire were added t dmsification. A thin emulsion is evaporated by spraying a pressure of 10 Torr for 3 hours, resulting in a transparent liposome solution. By anaizing the sample (chromatography on Sephadex G-50
It is established that the output of i Hp; ar; cyj; ir.o-panyl is 85% at black.
For example, K 3 MP dibutyl ether was added with 0.1 f-gl aqueous solution of 3, 9-bis-dimethyl; 1 of 1 azioti p-1H chloride chloride) 1 Trap1 and 0.001 M and 0.15 M lASS. and 55 mg dipalbmitoyl-foefat; -1dilcholine. mixture subjected; s} from ultrasound
effect, as described in example 1, and a transparent organic solution is obtained, containing a microbubble1) Ci (.p1 - sedimentary base) of the specified aqueous solution,
surrounding layer: dipalmitoyl-phos-fattide and T X o JH-T n a. The orga gus assortment is then centrifuged for 20 min at 8000 s ,. At the end of this stage, a transparent surface is obtained and a semi-transparent blue phase containing microbubbles. This phase, which has a consistency gel, is transferred to 10-mil.iroB1- ;; reservoir, where its mixes are supplied with plutgel help with 0.3 mp 200 mg / ml
solution, (july pnoe ratio 20; 1) mix 1 pglecithin and phosphatidylser -; ia in dibutic. simple, simple, then 5 are added to the solid mass, 1 L of 0.1 L of a clear solution of L-aCO and the mixture of 1 mixture is intensively mixed by mixing with a magnetic stirrer. After 101 5 min, a homogenous transparent solution is obtained, containing in suspension lstosomes with mixed shells. The outer layer of this cellulose consists of a mixture of granules: 1 and green phosphatidyls, and its core is 1; 1 dipole-mitoyl-phosphatide: 1H1; 11a.
Example 10: Liposomne precursors, withholding insulin, Poluch1yut as described in Example 1. Then, instead of directly using organic organic
a solution containing microbubbles in an aqueous 0.9% solution of l / aC (, the organic solution is first concentrated 1ot, subjected to reduced pressure1-: and: o (20 torr-) at room temperature
temperature After evaporation. organic solvents at the bottom of the tank receive an oily translucent 1 st: st layer consisting of adherent microbubbles. All this is added to the flask with 7 l of an aqueous solution of 0.9% and with the help of a magnetic stirrer this oily phase dispersion oil; are in the aquatic environment.
This oily phase gradually disappears and after 10–15 minutes a clear geyugenny solution containing the necessary liposomes in suspension is obtained. After chromatography of this solution on Sephadex 4B (Pharmacia Sweden), the analysis of the non-encapsulated phase shows that 52% of the original insulin is effectively encapsulated in liposomes.
Example 11 a A mixture of lecithin 40 g, 600 ml dibutyl ether and an aqueous 0.9% solution of l / aCg containing 10 g / l insulin, 400 ml is homogenized using a MIN1SONIC homogenizer (Ultrasonics Ltd, Great Britain) . Get a stable milky solution, which is poured in A-liter of a flask containing 2 liters of water 0.9%
solution / VaC. Then, using an emulsifying mixer, the organic suspension and medium are emulsified together for 15 minutes. After that, the organic solvent is removed by blowing air, i.e., the mixture circulates downward in the column opposite
FLOW subplme; of air. The latter is collected, after saturating the solvent with vapor, at the top of the column. In this way, a homogeneous translucent solution of liposomes containing insulin is obtained.
Example 12 3 ml Dibutlnaya EF11P, 9% aqueous solution / AC5- containing 10 mg / ml luliia, 1 ml and 125 mg Leaditine are placed in a solid 1O ml gyrex tube together with 2 g glass glasses of 1 mm diameter The tube is closed, placed on a stirrer and three days for 30 minutes at a speed of 100 rpm. In this way, a rather homogeneous milky solution is obtained, which is drawn into a T00-ml-1-flask flask containing 30 W 1 of an aqueous 9% solution. Then the organic solvent is evaporated, as described in Example 2, using a stream of nitrogen. After purging for 20-30 minutes, a lipid solution of liposomes is obtained, containing 11-silent; 1-capsuled insulin.

权利要求:
Claims (5)
[1]
1. METHOD FOR PRODUCING LIPOS, which involves dispersing the encapsulated liquid in the presence of phospholipids and a solution of a surfactant in a water-insoluble organic solvent using ultrasound and removing the solvent, which differs in that, in order to simplify the method, after dispersion to the resulting mixture add water or an aqueous solution of salts, emulsify with stirring, and remove the solvent by passing gas and distillation or distillation under reduced pressure before or after emulsification.
[2]
2. The method according to claim 1, characterized in that benzene, alkyl- and gas-alkylbenzenes, aliphatic esters, aldehydes, esters, ketones, free and halogenated aliphatic and cycloaliphatic carbon are used as a water-insoluble solvent s.
[3]
3. The method according to claim 1, characterized in that the lipids use substances from the group of lecithin, phosphatidyl-ethanolamine, lysoleiitium, lysophosphatidyl-ethanol-amine, phosphatidylserine, phosphatidyl-_nositol, sphingomyelin, cardioli - 3 pins, phosphate acid, cerebroside, stearylamine, dipalmitoylphosphatidylcholine or a mixture of one phospholipid, cholesterol and tocopherol.
[4]
4. The method according to p.
[5]
5. The method according to claim 1, characterized in that dyes, aromatic substances, NyAA e O ehe drugs, enzymes or polypeptides are used as the encapsulated substance.
1 1 580'31

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法律状态:

优先权:

申请号 | 申请日 | 专利标题

CH961677A|CH624011A5|1977-08-05|1977-08-05|

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