CA2041075A1 - Process for the production of aqueous liposome suspensions containing active ingredients - Google Patents
Process for the production of aqueous liposome suspensions containing active ingredientsInfo
- Publication number
- CA2041075A1 CA2041075A1 CA002041075A CA2041075A CA2041075A1 CA 2041075 A1 CA2041075 A1 CA 2041075A1 CA 002041075 A CA002041075 A CA 002041075A CA 2041075 A CA2041075 A CA 2041075A CA 2041075 A1 CA2041075 A1 CA 2041075A1
- Authority
- CA
- Canada
- Prior art keywords
- process according
- active ingredient
- liposome
- aqueous phase
- solution
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Abandoned
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/10—Dispersions; Emulsions
- A61K9/127—Liposomes
- A61K9/1277—Processes for preparing; Proliposomes
Abstract
Abstract of the Disclosure A process for the production of aqueous liposome suspensions containing active ingredients from a solution of a substance (substance mixture) for forming liposomes, in a lower alcohol with at most 3 carbon atoms and an aqueous phase, an active ingredient or active ingredient mixture is dissolved or suspended in the lipid solution and/or the aqueous phase. The aqueous phase is added to the alcoholic solution with mixing, the lower alcohol is removed by vacuum distillation and the liposome suspension is optionally freeze-dried.
Description
PR~CE:8~ FOR T~IE P~ClDUCq! I:~N O~ A~VEOtJ~ O~ 3~3PEN~3IO~
CO~l!Al:lilIN~ AC~ N~ )IENq!B
.
~ y Q~ ~h~ l~v~io~
The invention relate~ to ~ process for the production o~ aqueous liposome suspensions containing active ingredient~ from a ~olution o~ a substancQ or. a mixture of substance ~also desi~nat~d b~low ~ lipid or lipid mix~ure), which ~onm liposomes in a lower al~ohol with at most 3 aarhon atom~ and an aqueou~ phase, the acti~e in~rediant or the active ingredi~nt mlxture is d~'s~lved or ~u6pe~ded i~ ~he lipid ~olution andJor the a~ue~us pha~, characteri~ed in tha~ ~he a~ueous pha~ is added to the al~oholic solution with homogenizati~n, and the l~wer alcohol is removed by vacuum distillation.
As is ~nown, liposomes are sel~-contained spherical or e~liptical lipid ve~iale~, whi¢h enclose an ~qu~ou9 phase.
Depe~ding on the numb~r of lipid double layeræ pre~ent, large and small unilamellar liposome~ (LW and S W) are distin~ui~he~ from multil~mell~r l~pos~ tM~V).
The fir~t lipo30mes wer~ pro~uced by ~ispersion of lipid films i~ agueous ~hases. The MLV di~per~ions ~hu~
obtained aan b~ aon~erted to S W ~y u~ing ~uitabla homogeniza~ion proce~s~s, ~uch a~, e.g., ul~rasonic irr~diation or high-pres~ure homogeniz~tlon. But M~V and S W exhibit only a small inaluslon volume (volum~ o~
F~ lr,~ .t, ;~ EI ~ r~ 122 P~
7 rJ
CO~l!Al:lilIN~ AC~ N~ )IENq!B
.
~ y Q~ ~h~ l~v~io~
The invention relate~ to ~ process for the production o~ aqueous liposome suspensions containing active ingredient~ from a ~olution o~ a substancQ or. a mixture of substance ~also desi~nat~d b~low ~ lipid or lipid mix~ure), which ~onm liposomes in a lower al~ohol with at most 3 aarhon atom~ and an aqueou~ phase, the acti~e in~rediant or the active ingredi~nt mlxture is d~'s~lved or ~u6pe~ded i~ ~he lipid ~olution andJor the a~ue~us pha~, characteri~ed in tha~ ~he a~ueous pha~ is added to the al~oholic solution with homogenizati~n, and the l~wer alcohol is removed by vacuum distillation.
As is ~nown, liposomes are sel~-contained spherical or e~liptical lipid ve~iale~, whi¢h enclose an ~qu~ou9 phase.
Depe~ding on the numb~r of lipid double layeræ pre~ent, large and small unilamellar liposome~ (LW and S W) are distin~ui~he~ from multil~mell~r l~pos~ tM~V).
The fir~t lipo30mes wer~ pro~uced by ~ispersion of lipid films i~ agueous ~hases. The MLV di~per~ions ~hu~
obtained aan b~ aon~erted to S W ~y u~ing ~uitabla homogeniza~ion proce~s~s, ~uch a~, e.g., ul~rasonic irr~diation or high-pres~ure homogeniz~tlon. But M~V and S W exhibit only a small inaluslon volume (volum~ o~
F~ lr,~ .t, ;~ EI ~ r~ 122 P~
7 rJ
enclosec~ aqueou~ pha~3e [ lit~,r/mol lipid] ), 30 that ~hey are not suitabl@ ~or ef~ective inclu~ion of hydrophilia sub~:tances ~
In contr~6t, LW exhibit a clearly in~rea;ed inclu~ion volume and thu~ an .tncreased ~n~lusion capacity ~meaf~:ured on the ~asi~ o~ the ~6 of th~ total active ingredien~ that can possibly be aontain~d in the inclu8ion vslume) . LW ' s hav~ a partlale ~l~e of 100-10, 000 nm, preferably 200-2, 000 nm.
Tha most common method~;, up to this day, ~or production of LW C:~n be ~ummarize~ under the generic tex~n o~ olvent e~rapora~ion" methods .
The method prol~ably ~c~st known and beæt wit~ re~;pec:t to the ~ nqlu6ions obtair~ed i~ the RE~ (rever~;e pha~e evaporatiorl) method o~ Papahadjopoulo~ ~U.S. paten~ no.
4,23~,871). In this ~::a~3~3i th~ lipid or lipid mixtur~ i~
fir~t di~;~olved in a ~olv~nt whi~h is not, ~r only 31ightly, w~ter-miscible (g~ner~lly, diethyl ether or chlorofor~
After additic~n to the aqueous phase containing pharmaceutical ~ub~tanc~e~, the mixture is then cor.verted to a W/0 emulsion ~water-~n oil emul~iun) by ultrasonic:
irradiaticn. The solvent i~ then remov4d ~rom th~ latter u~ing a rotary e~apora~or, and a ~1 is fir~t ~o~ d which is converted to a lipo~ome suspenRion by increa~ing ~h~
vacuum or add ing water ~
In contra~t to thi~, in the 'lether in~ection" method introduced by D~am6!r ancl Bangham [13iochf3m. ~3iophy5. Acta, 443 ~1976) 62~-~34], the lipid, also di~olved in diethyl ~ther, is injecte~l in~o a warm aqueouR solutian (50-60C) under partial vac:uum. ~he lipo~omes oh~ain~d a ter filtration through ~ 1. 2 micron filter ar~ heterogen~ous~ and exhibit ~ize i; ~etween 150 250 n~a~
~he solver~t~ u~ad ~n the:e ~Aethods are to b~ viewed critically Por tox~ aologlcal and ~af~ty xea~3on~c .
/ u ~ ~3 But, if ethanol is used a~ a solvent in ~ proce~
preoedlng ~he "ether inj~3ction" method/ greatly diluted SW
d.ispersion~ result which ~re concentrate:d by ultr~filtration ~Bioahem~ Biophy6~ Act~, 298 (1973) 1015-1019~. The ~vera~e $ ~l~e of th~3 liposom~3 thu~2 oktain~d is ~learly under 50 nm and the MLV portion i9 lndicatecl a~ 6%.
An e~odiment of thi~ proc~ i descri~ed in ~5P-A 0 253 ~l9. In ~hiY proce~ or exampl¢, an ethanolic lipid and ph~rma~eu~iaal ag~n~ solul;$on, which constitute~ a tot~l of 10% o~ the to~al formulation, ~re inject~a under pre~ura o~ 1,000 to 3,000,~00 h Pa (heato Pas~als) in an agueou~ pha~e, wh~.ch is agitated by a homogeniz~x.
According to this proa~s~, ~7hi~h $~ very ~xp~n~ive, sm~l~
unilamell~r llposomes r~ult. ~hi3 proce ~ i~ unsuitabl~
fo~ production o~ liposome ~uspensions whi~h cc~nt~in hydrophili~ active ingredien~ .
Also workh m~ntioning i8 a method for the p~oduction of liposome~ with water-~olu~le ~olv~nt~;, whi ::h include~ the production of a "monophase" ~Wo 85/00751). Thi~ ona-pha~o 2~ mixture ic gen6~rally produa~d ~xom 5 ml o~ th~3 lipid-conta ~ nlng solv~nt ( ~ . ~ ., ethanol ) and 0 . ~ ml o~ the ~ ous component. ~h~ ~olvent i~; then removed by introducing an inert ~a~ duxin~ ~he ~imultaneous ultrasoni ;: irradiation o~
the mixture, and a ~ilm r~sults. ThQ latter i~ then ~5 re~;u~pended with an a4ueous ph~ e. In thi~ ca~e, so-calle~l ~: MPV (monophasic ve~i41e~) ~esult, which exhibit ~ever~l lipld doubl~ layers. R~la~ive to the ~t~ndard MPV, the MPVs produced by the ~bov~-d6~scribed process exhi~it incr~a~ed tabil~y in buffers a~ well as incr~3a~ed 3 Q incluxions .
Final}y, the proaeo~ de~cribed in EP-A O 349 429, in whlah an ~thanolic lipid solution optionally ~on~ln~l~g the active ingredient i~ in~roduc~d into an ~queou~ pha~e w~ th 7^~
In contr~6t, LW exhibit a clearly in~rea;ed inclu~ion volume and thu~ an .tncreased ~n~lusion capacity ~meaf~:ured on the ~asi~ o~ the ~6 of th~ total active ingredien~ that can possibly be aontain~d in the inclu8ion vslume) . LW ' s hav~ a partlale ~l~e of 100-10, 000 nm, preferably 200-2, 000 nm.
Tha most common method~;, up to this day, ~or production of LW C:~n be ~ummarize~ under the generic tex~n o~ olvent e~rapora~ion" methods .
The method prol~ably ~c~st known and beæt wit~ re~;pec:t to the ~ nqlu6ions obtair~ed i~ the RE~ (rever~;e pha~e evaporatiorl) method o~ Papahadjopoulo~ ~U.S. paten~ no.
4,23~,871). In this ~::a~3~3i th~ lipid or lipid mixtur~ i~
fir~t di~;~olved in a ~olv~nt whi~h is not, ~r only 31ightly, w~ter-miscible (g~ner~lly, diethyl ether or chlorofor~
After additic~n to the aqueous phase containing pharmaceutical ~ub~tanc~e~, the mixture is then cor.verted to a W/0 emulsion ~water-~n oil emul~iun) by ultrasonic:
irradiaticn. The solvent i~ then remov4d ~rom th~ latter u~ing a rotary e~apora~or, and a ~1 is fir~t ~o~ d which is converted to a lipo~ome suspenRion by increa~ing ~h~
vacuum or add ing water ~
In contra~t to thi~, in the 'lether in~ection" method introduced by D~am6!r ancl Bangham [13iochf3m. ~3iophy5. Acta, 443 ~1976) 62~-~34], the lipid, also di~olved in diethyl ~ther, is injecte~l in~o a warm aqueouR solutian (50-60C) under partial vac:uum. ~he lipo~omes oh~ain~d a ter filtration through ~ 1. 2 micron filter ar~ heterogen~ous~ and exhibit ~ize i; ~etween 150 250 n~a~
~he solver~t~ u~ad ~n the:e ~Aethods are to b~ viewed critically Por tox~ aologlcal and ~af~ty xea~3on~c .
/ u ~ ~3 But, if ethanol is used a~ a solvent in ~ proce~
preoedlng ~he "ether inj~3ction" method/ greatly diluted SW
d.ispersion~ result which ~re concentrate:d by ultr~filtration ~Bioahem~ Biophy6~ Act~, 298 (1973) 1015-1019~. The ~vera~e $ ~l~e of th~3 liposom~3 thu~2 oktain~d is ~learly under 50 nm and the MLV portion i9 lndicatecl a~ 6%.
An e~odiment of thi~ proc~ i descri~ed in ~5P-A 0 253 ~l9. In ~hiY proce~ or exampl¢, an ethanolic lipid and ph~rma~eu~iaal ag~n~ solul;$on, which constitute~ a tot~l of 10% o~ the to~al formulation, ~re inject~a under pre~ura o~ 1,000 to 3,000,~00 h Pa (heato Pas~als) in an agueou~ pha~e, wh~.ch is agitated by a homogeniz~x.
According to this proa~s~, ~7hi~h $~ very ~xp~n~ive, sm~l~
unilamell~r llposomes r~ult. ~hi3 proce ~ i~ unsuitabl~
fo~ production o~ liposome ~uspensions whi~h cc~nt~in hydrophili~ active ingredien~ .
Also workh m~ntioning i8 a method for the p~oduction of liposome~ with water-~olu~le ~olv~nt~;, whi ::h include~ the production of a "monophase" ~Wo 85/00751). Thi~ ona-pha~o 2~ mixture ic gen6~rally produa~d ~xom 5 ml o~ th~3 lipid-conta ~ nlng solv~nt ( ~ . ~ ., ethanol ) and 0 . ~ ml o~ the ~ ous component. ~h~ ~olvent i~; then removed by introducing an inert ~a~ duxin~ ~he ~imultaneous ultrasoni ;: irradiation o~
the mixture, and a ~ilm r~sults. ThQ latter i~ then ~5 re~;u~pended with an a4ueous ph~ e. In thi~ ca~e, so-calle~l ~: MPV (monophasic ve~i41e~) ~esult, which exhibit ~ever~l lipld doubl~ layers. R~la~ive to the ~t~ndard MPV, the MPVs produced by the ~bov~-d6~scribed process exhi~it incr~a~ed tabil~y in buffers a~ well as incr~3a~ed 3 Q incluxions .
Final}y, the proaeo~ de~cribed in EP-A O 349 429, in whlah an ~thanolic lipid solution optionally ~on~ln~l~g the active ingredient i~ in~roduc~d into an ~queou~ pha~e w~ th 7^~
light ~t~rring, is al~o to be mentioned. A~ our tesk~ ~how, conslderably l~w~r inclu~ion c~p~cit~ are a~hle~ed ln this prev~ously known proce~s than by the proce3~ aacor~ing to the invent~on. Moreover, the process acaording to the invQntion has ths advantage th~t with it~ help, even with lipid co~centration o~ over 10% rel~ive to the e~hanoli~
phas~, high inclu ion capacitieæ or even a ~urther increase o~ ~he inclusion capacities ~an b~ achi~v~d.
In comparison wlth these previously known methods, the proces~ ~ccord~ng ~o the invention has ~he advant~g~ ~hat it i~ technically feaci~le on a large s~a~e in a simple wayA
Th~ u~ of strongly toxic or very ea~ily ~lammabls -~olvents i~ avoided. ~The ~ery ~ood reproducibility o~ tha pro~s~
according to the inven~ion i~ also ~o ~e empha~ized relative to the obtained a¢ti~e ingredient in~lu~ions ~nd li~osome -~izes, as well as the very hi~h active ingredient inclusion in the liposomes, up to over 50~, and the unu~ually high ~ctive in~redient ooncen~ra~ions, obtained ~y ~he proce~s accord~nq to ~he ~nven~ion, in the liposome 6usp~n~ion~ ~up to 800 mg/ml). Active ingredient inclusion means the percentage of the total active ingredient ~mployed which is contained in the inclusion ~olume of the lipids. ~ur~her, it is worth m~ntioning that it i~ possible to produae liposQme su~pensions, which have only a very l~w r~idual solvent cont~nt and which hav~ a good shelf li~e, with the help of the proc2s~ aocording to the invention. The proces~
ac~ording to the invention can al~o ~e per~ormed ~robl~m-fre~ under a~ptic conditions.
To perfor~ the proce~ according t~ th~ invention, ~he same substances for forming liposom~s can be used ~s in the previou~ly lcnown proce~-~es~
Sub~tanc~s ~orming ~uitable lipos4mes are u~ually amphiphatic lipid~ oX lipid mlx~ure~, ~uch a~, ~or ~xample, 2 ~ 7 ~
pho~pholipid~, sphin~omyel~nY or eth~r pho~phollpids. ~hese lipids aan h~ve straight-oh~ln or branched, ~aturat~d or un~aturated, similar or dif~erent acyl ~id~ chains.
Sult~ble phospholipld~ are, ~or example, the pho~phatldylchol~ne~, the phosphatidylethanolamine~, the phosphatidylserines, the pho~phatidylinosikol~, the pho~phat~dylglycerols or ~he pho~phatidia ~cids~ Sultable ether pho~pholipid3 are, for example, the plasmalogens (Dr. Otto~Albert Neumuell~r: Roempp~ C~2mi~-Lexikon;
Franak~che Verlagshandlung, Stu~tgart ~D~) 2665, 159, 3g20 and 4045).
For the production o~ liposom~s, lipids or mixtures thereo~ and in particular al~o mixtures of the~e ~ipid~ with cholesterol, chole~terol hemi~uooina~e, alpha-tvcoph~rol, alpha-tocopherol h~misuccinate and/or charg~ aarriers, such a~, for example, stearyl amine, ~teario ~aid, diethyl phosphate, oleic acid~ palmitic acid, bile acid, Cuch a~, for QXample, choli~ a~ld, glycocholi~ a~id, ~an ~e us~d.
Suitabl~ mixture~ can contain about up to ~0 mol~ parc~nt o~
cholester~l an~ up ~o ~0 mole peroent ~f charge c~rri~r. As solvent ~or the phospholipid3 or miX~ureR~ pre~erably isopropanol or in particular ethanol is ussd.
In principl~ the proa~ss aacording to the invention should al~o be feasible with water-~oluble low-boiling solvents o~her than lower a~cohols: for exampl~
t~trahydrofuran is suitable. But, performance of thi~
prooes~ embodiment i5 ~onsiderably ~ore expensiv~.
T~ perform the proce~ ~ccording to the invention, alcoholic ~olutt~n~ are pr~Qrably u~d which contain 0.
to 30 g of lipo~ome-forming ub~tance or su~ance mix~ure per lOO ml o~ ~olvent~ I~ nece6~ary~ the~e ~olutions are produ~ed by he~ti~g the components.
HPF~ i r~ .. lr~ I r~ _E! ,i~ l rEI_ 1 0-l:, i~ P0-~
~ ~ ,1, s Th~ proQass accerding to the invPn~ion i~ p~r~ormed, a~
wa~ already menti~ned, in such a way that the aqueous phase i~ added to ~he alcoholic solution with mlxin~ (such ~, for ex~mple, vigorou~ ~irrlng) and t~e lower alaoh~ re-moved, ~or exampl~, by vacuum di~tl llation or by introducingnitrogen. The mixing rat~ is g~ne~ally 50-1000 rpm~, pre-~e~a~ly 100-500 rp~.
Optlonally, th~ ~ispersion ob~ained after ~ombining the a~u~ou-~ pha~e and the al~ohollc 6ulution i~ mixed ~or so~e 10time ~about 10 to 1~0 minutes~ at a temperatura o~ about 20 to 90C, be~ore the solven~ is compl~tely or parti~lly sepa rate~. The volumstric ra~io of a~ueou~ phase ~o lipid pha~e is generally abol~ 50-1000 ml, pre~erably 100-500 ml o~
aqueou6 pha~e per lQ0 ml o~ lipid phase.
15The temp~ratur~s uit~ble ~or ~he proceY accordi~g to th~ invention are depend~nt on ~he solubility ~ th~ lipo-~ome-forming sub~tances ~ well a~ ~heir pha~e tran~itl~n temperature ~t~ he h~a~ stability of the activa ingr~-dient or a~tive ingr~dient mixtu~e ~nd ~h~ vapor pre~sure o~
~he alcohol to be ~epa~ated. ThP tempera~ure i6 pr~f~rably ~etween 20C and 90~C ~nd in partiaul~r b~tw~n 40~C and 70C. In general, it will be su~ ient to oper~te w~th a v~cuum of 10 h Pa to 100 h Pa.
The aqueou~ pha~e used ~or the pro¢es~ ac~ording to the in~ention can op~ionally ~ontain bu~er substan~e~ and/or i~otonizin~ add~ti~R~ Suitabl~ additl~es are, for example, inorganic or organiC ~alts or bu~fer subs~an¢e6, such ac 60dium chloride, ~ris buf~er, pho~pha~s buf~er, a~tr~t~
~uff er, glycine ~uf~r, citrate-pho~pha~e buffer, male~te bu~r, etc. Mono~ ~ di~ charides, ~uah as glu~ose, la~to~e, s~harose or trehalose, ~ugar alcohol~ ~u~h ~6 manni~ol, or~itol, xylltol or ylycerine or wate~-~oluble polymer~, ~uch as dextran or polyethlyene glycol.
Sinc~ th~ lipid~ and ~lso s~veral activ~ in~redi~n~s are se~itive to oxidation, th8 a~ue~u~ phas~ Gan be mixe~
with antioxldan~s, suoh s sodium a~cor~ate, tocophQ~ol or sodium bisul~ite.
The a~ueous l~posome susp~nsions produced aaco~d~ng to the proaess o~ the invention are used preferably ~or encap~ulaking water-~oluble actiVQ in~redi~nt3.
Suah water-soluble aa~ive ingr~dient~ are, ~or example, diagnostic agent~, such as the X-ray contr~t ~dl~
- lotrolan, iopromide, ioh~xol, iosimide, motriz~mide, s~lt~ of ami~oa~etic acid, iotroxic zcid, lop~midol, 5-hydroxyacetamido-2,4,6-triiodo-i~ophthalic acid-(2,3-dihydroxy-~-methylpropyl)-t~hydroxyethyl)-di~mide (=ZK
119095) and 3-carbamoyl-5-~N-(2~hydroxyethyl)-acetamido~-2,4,~-triiodo-benzo~ acid-~(lRS,2$~ ,3-dihydroXy-l~
hydro~ym~thylpropyl]-~mide (-ZX 13~129) or NM~ contra msdia, ~uch a~ gadolinium DTPA, yadolinium ~OTA and the gadolinium complex o~ 10-~1-hydroxymethyl-2,3-dihydroxypropy~ 4~7-~rls-t(carboxy~thy~ 4~7 tetraazacyclodecane~
Suitable ther~peutic aetive ingredients are, among others, antibiotio agents; such as gentamyoin or kanamycin, aytostatic ag~nt~, such a~ doxorubiain hydroohloride or cyclopho~pham~de and virustat~c ~ge~t~, such as ~idarabin~
or active ingredients, suah as mitoxantrone hydro~hloride~
These water-soluble active ingredi~nt~ ar~ dissolved in th~ a~ueous phase b~or~ per~o~mance o~ he proce~
according to the invention.
Furth~r, the ~queous liposoma susp~nsions aan ~l~o ~e US~d to encapsulata sli~h~ly soluble act~e ~ngr~dients in water.
Such active ingredient~ are, ~or example, plant protecting agents, such a~ ~lightl~ soluble inR~cticides or herbicid~s ~nd in ~ icular ~lightly oluble pharm~eutia~l active ingredient~.
,3~
Sligh~ly water-soluble or insoluble pharmaceutical active ~ n~redlents of the :~ollowing actlve ingredient group~3 are suitablet ~or example, for production o~ aqueous su~pen~;ion~ according to the proae~s cs~ the invention.
Ge~agenlcally a~:t$~e 6terc~id hormone~ uch a6, for ~xa~ple, 13-ekh~ 7~eta-hydroxy~ s-d1nor-l7alpha-pr~gn-4-en-20~ 3~ne ~levonorgestrel)~ 13-e~h~1-17be~a-hydroxy-18,19-dinor-17alpha-pregna-4,15-dien-20-yn-3-one (-ge~to~en) or 13-ethyl-17~e~a-hydroxy-11-methylene-18,1g-dinor-17alpha-1~ preqn-4-~n~20~yn (=de~oge~trel).
Estrogenally a~tive steroid hormones, such a~ 3-hydr~xy-1,3,5-(lo~-es~ra~rlen~17-on~ (-e~one~ or l,9-nor-17alpha-pregna-1,3,5~10~-trien-20-yn-3,17beta-diol ~thinyle tradiol).
Andro~ni~ally aati~ ~teroid hormone~, ~u~h a6, for example, 17~eta-hydroxy-4-andros~en-3-on~ (-t~sto~terone) and i~ e~ter or ~7b~ta-hydroxy-lalpha-methyl-5Alpha-androsten-3-on~ ~-me~terolone).
Antian~rogeni~ally ~c~i~e ~tero~d hormo~e~, such as, ~or example, 17alpha-aaetoxy-~-chloro-lbeta~2~eta-dihydro-3H-ayclopropa~l~2]-pregna-l~4~6-triene-3~2o-diono ~cypot~ronac~tat~)~
Corticoids, ~uch a~, for example, llbeta,17alpha,21-trihydroxy-4-pregnene-3,20-~ione (=hydroaortison~), llbe~a,17alpha,21-trihydroxy-1,4-pragnadi~ne-3,2~-dinne t=prednisolone), llbeta,17~1pha,21~trihydroxy~6alpha-methyl-1,4-pr~gnatriene-3,20-dione (-methylprednisolone) and 6~1pha-fluoro-llbeta,21 dihydroxy-l~alpha-methyl~1,4-prqgnadien2-3,2Q-dione (~ifluoortolone~ and their est~r~.
Ergolines, such as, ~or example, 3-~9,10-dihydro-6-methyl~8alpha-er~olinyl)-1,1-diethylure~ (=ergoline~
~romo-g,10 dihydro-6-mqthyl-8alpha-~rgolinyl~
3 ~
_ g ~
diethylurea ~=bromergoline) or 3-(6-methyl-8~1pha-ergolinyl)~ die~hylurea (=ter~ride).
Antihyper~e~sive agents, such as, for example, 7alpha-~cetylthio-17~1pha-hydroxy-3-oxo-4-pregnen~-21-carboxylic acid-gamma-laotone (aspironolactone) or 7alpha-ac~tylthio-15beta~1~bet~methylene-3-oxo-17alpha-pregna-1,4-diane-21,17-carbolactQns (~m~spirenone).
Antiaoagul~nt~, ~uch a~, f~r example, 5-[hexahydro-5-hydroxy-4-~3-hydroxy-4-methyl-1-octen-~-ynyl)-2(1H)~
1~ pentalenyliden~ pentanoi~ acid ~-iloprost~.
P~ychopharmacological ayents, suah a~, ~or exampl~, 4-(3-oyalopen~yl~xy-4-~ethQxy-phenyl-2-pyrrolidone ~rolipram) ând 7-chloro-1,3-dihydro-1-m~thyl-5-phenyl-2H~
1,4-bQnzodiazepin-2-one ~-diazepam).
Carotinoid~, auch a~, for example, ~lpha-oarotin and bata-carotin .
Fat-solubl~ vitamin~, ~uch a~, ~or example, vitamlns the vitamin A, vitamin ~, ~itamin E and vitamin K group.
Beta-oarbolines, as they ar~ de~cribad, ~or ~xample, in European patent appl~cation~ 234,173 ~nd 23~ 7, aro another group. A~ beta-carboline~, ~or ~xampl~, there can be mentloned 6~be~zoyloxy-4-m~thoxymethyl-b~ta-~rboline-3-carboxylic ~cid-isopropyl ester ~=b~carnil) and 5-~4-chloroph~oxy)-4-methoxymethyl-~eta-carboline~3~aar~0x~
a~id-1 opropyl est~r ~=Cl-P~O~IP).
Sligh~ly ~olubl~ or lnsoluble oon~ra~t m~dia are al80 worth mentioning, such a~ the X-ray contra~t medlu~
iodlpaml~e ethyl ester or NMR contrast media, such a~ the iron or manganes~ porphyrin chelates or ~l~o the ~gnetit~s.
As ~uitable active ingr~dients, salic~lic ~id, re~noic ~cld and azalaic acid further can be mentionqd~
Anti~y~otin~, such a~ the amphotericin B, ~conacol or meconazol ~an al~o be used.
-- 10 -- .
Before par~or~ance o~ the proce~3 aca6rding to the invention, th~:e aotive in~redient~3 ara disE;olved or su~:pended in the a}coholic solut:ion or ln the aqueous pha~e.
In the proces~ ~ccording to th~ invention, generally 0 . 05 to lo g and preferably 1 ~o 3 g o~ ac:tlve ingredient ls used per g o~ ~ubsta~nc~3 ~subs~anc6 mlxture) for forming liposom~. S~coril~ liposome suspensions can ~e produced in a ~i~nple way by the prooess ~coording to the inv~n~ion by both solutions being sterili7.ed by :Eiltration be~ore 10 per~ormanc~ o~ ~he proc~ss and all ~ubse~uent prooess 8~ep6 being per~ormed under a~eptic conditions.
~h~ inclu~ione~ which are achievable in the ll:posome ~u~pensions are dependent, o~ aourss, on tha typf3 o~ active in~redient and the substance ~ub tance mixture3 ~or ~orming 15 lipo om~a~a a~ w~ll a~ their ratio to one another.
Optionally, the unencap~ulated ~ctive ingredlen'c can ~e r~mo~rad, ~or examplo, by ul.trafiltxa~ion, dialy~
micro~iltration or ~entri~uging.
Tha lipo~ome ~uæpension obt~lned c:an be dl.rectly 20 stored. Alternatively, the llpo~ome su~pension can be converted by freeze-drying ~o a stable fo~m o~ ~orag~.
Be~ore the freeze- drying, if neoessary, among ~ther~, ~or example, additi~res, suah as mannitol or ~orbit~l and/or elec~rolytes and vis~osi~y-ln~luencing a~e~t~ su~h as sodium 25 chloride aan be added to the material~ to ~e fr~ez~-dried.
The lyophilizate~ can b~ re3usp~nded with bidistilled water or a~ueou~ phases, which ~an con~ain the same a~ditives aa th~ aqueouæ pha~e~ u~ed ~or lipo~ome prsduation.
Ths amount of r~suspQnsion m~dium can in this c~se b~
O.S t~ 20, but preferably 1 to 6 ml per g ~f lyophiliza~.
The ~u~pen~lon~ obtained can be uaed dlreatly or after ~iltration through filters of ~uitable pore ize.
2 ~ 7~
~ 11 ~
The lipo~ome~ produced by the proc~ss aecordlng to the ln~en~io~ ~.an, for example, b~ used i~ they contain ~ontr~3t media for N~R or X-ray dia~nosis in various diagno23~ic proc~s~s. ~hs~e include, for example, the di~gnosis of tumors in organs of the reticulo-endo~hellal ~ystem (~or example, liver and ~pleen~ or other i~trava3cul~rly acc~ible ti~3ue6 as well as the arthrography. The extrava~cular admini~ra~.lon o~ su~h lipo60~e ~usp~n~ion~
(such a~, ~or sxample, subcu~aneou~, intramuscular or intraperitoneal) ~an al~o be use~ ~or dia~no~is purpo3~s, ~uch as, for example, the indirect and ~irea~ lymphography, or for ther~pautl~ purpose~ ~such a~ intramu~cular depot prepara~lo~. Of c~ur~e, it i~ al~o po~sible to admin~ter tha preparations orally, and the prPparations opti~nally ~an be filled in aoat2d cap~ule~ rasist~nt ~o ga~tric jUiC~5.
Further, the liposom~ su~pensions containing active ingredients produaed ~y ~he pro~ aoco~ding to the invention can be applied in the ca~e o~ suita~le lipid compo~ition as blood pool ag~nt~ or a~ ~arget-directed pharm~c~utical vehicles.
Without further ~la~oration, it 1~ bel~eved ~hat one ~killed in the ~rt can, u~ing th~ pr~a~ding d~scription, utiliza the pr~sent invention o its ~ulle~t extent~ The ~ollowlng pre~erred speci~ic emhodiments are, ther~ore, ~o ~5 be construed a~ merely ~llu~tr~tlve, and not limitative o~
the remaind~r of the disclosure in any way whatsoever.
In the ~oregoing as~d in th~ ~ollowing ex~mples, al~
temperature~ are ~et ~o~th uncorrected in degree~ Celsiu~
and unle~s otherwiæe indioate~, all parts and peraentage3 are by weight.
The entir~ disclo3ure~ o~ all appli~ations, pat~nt~ ~d pu~lication~ ted ab~e and below~ and o~ corr~ponding 2 ~ 7 .j ~ 12 ~
appliaation Federal ~epublic o~ Germ~ny P 4~ 13 580.2, ~iled ~p~il 24, 19~0, are her~by incorporated by re~r~nce.
In the ~ollowing exampl~, th~ foll4wing ~breviation~
are u~ed:
PC - Ph~sphatidylcholine S 100 o~ the Lipoid KG
comp~ny CH = Powd~red ~hol~terol ~JP) ~ ~he Merck AG
company SA V~ry pure st~aric acid oP the Flu~a AG
company ~PPG = Dipalmitoylphosph~tidylylyaerol DPPG o~ th~
Lipoid KG company HSPC =~ Hy~rogenated 60yabean lQcithin ~Phospholipon 90H~ of the Na~t~r~ann AG company DCP ~ etyl phosphat~ of th~ ~igma company .
:
` `~
9.2~ g of lipid ~nixture (PC/~H/SA -- 4:5:1 mol:mc~ mol) is di~olved in loO ml ~ eth~nol and ~t~rllizad by ~iltrativrl at 70C. Th~n, th~ ~olu~iorl i~ trz~ns~rr~d to a reactiorl ve~6el he~ted moder~tely to 55~ and it i~ mixed with f3tirring (300 rp~ wi~h an ac~ive lngredient solution ~terlliæed by flltration (~ontaining ~7 . 75 g c~ ~ opromid~
~nd 200 ml o~ aqueou~ 20 mmol t:ri~ Cl b~fers pH 7~5~
lo T~en, th~ alcohol i~ ~listill~d of~ at 554C~ a vacuum ~I 5, 000 Pa and the re~ulting liposome su3pen~ion is axamined ~or its propertie3.
Then, the lipo~;ome su~pension i~ bottled dir~atly in portion~ of ~0 y in 50 ml gla~s in~usion bottl~ and it i8 ~r2~zQ-drisd. Th~ lyophillzates obtairled are re~usp~3nded 50 that an ~c:t~ve ingredient soncentration o~ a~out 2 oo mg/ml re~ults and al~o examined.
~L:L
Parformance o~ the te~;t talce~ place a; de3ar~ bed in ~0 exzlmpl~ xcept tha~ 18 . 5 g o~ iopromide is us~d.
Per~ormanc2 ot~ the ~ect takes pîac~ a: des¢ribe~ in exampls 2, except that 13 . 9 g of lipid mix1;ure i~ us~3d.
~xample 4 2~ P~arformancs of the te3t take~ placa as ds~cribed in ex~mple ~, except th~t 18 . 5 g af lipid mixture is u~ed~
-- 14 -~
~Ia S
P~rfor~anoe o~ ~h~ test took pl ace as desaribed in exam~pl Q 2, ~xcept that iotrol an i~; u~ed in~te~d of iopromide .
5 ~AP~ 5 Performance C)f the ~est takes placa a~; d~scri}:~ed in example 2, f3xc~pt that iopamidol is u~ed in ~cead o~
iopromide.
a~m~l~ .7 PerformanGe of tne te~t ta~es place as d~cribed in Rxample 2, eXc~pt that iohexal i~ used in~tead ~f i~promide.
~ca~pl ~3 ~
Per~o~ma~ce of ths t~st ~akes place a descri~ed in ex~mpl~ 2, ~xc~pt that the nonionic contra~t medium 5-hydroxyacstamide-~,4,~-tr~iod~ oph~h~lia acid-~,3-dihydrQ~y-N-methylpr~pyl~(2~hydroxy~thyl)-di~:ide ~ used in~tead of iopro~ide.
~x~mpl~ g Performa~ce of the ~e~t took pl~ce a~ desaribe~ in ~0 example 2, eXa~pt tha~ ~ lipid mixture of PC~C~/S~ 4:~
mol :mol :mol is used~
Per~ormanae o~ the taBt t~kes plac~ as des~ribed i~
example 2, except that a l~pid mixture of PC~C~/~C o~ 4:5:1 mol:mol:mol is u6ed.
J
~P~
Performance o~ the ~e~3t takes plaae ~ describ~d in exampl~ 2, exoept that a lipid mixture of PC/CH/SA o~ 6: 3 :1 mol: mol: mol i8 u~ed .
~ 2 Per~ormanc~ oi~ tha te~t take~ place a de~cri~ed in example 2, exc~pt that a lipid mixture~ o~ PC~C~/D~P of 4: 5 :1 i8 used.
Per~ormance o~ the te~t takes place as desc:ril:~e~ in example 2, exsept tha~ a lipid mixture o~ PC~C~I 1 s 1 is uaed .
Per~ormance o~ ~he te~ ~akes plaae as de2;crib~d in example 2, exc:ept that a lipid mixture of Pt::~HSPC/C~I/SA o~
2:2:5:1 i3 used.
Performance of the te~t take~ place as descri~ed in example 2, except that the batch size was ill~rea~d tenfold~
2 0 Per~ormanc:e o~ the test ~ake~ plac:e as descrihed in example ~, exc:ept that ~ one hunclred and ~ifty~old large~
batch 18 UBed.
~he followin~ table ~how~ the result~3 re~a~ed in examples 1 ~o 1~.
,, n, i7 ~.j . .
~;~X~
v ~ v ~ ~ ~ ~ ~ .
n ,. ~ <~ n ~ . g~ ~.
<~ ~ ~ ~ ~ ~ n ~ ~ ~ . ~ 1~
~ I ~ X ~ O 1 ~-V~ o ~ -o u~ ~ ~n ~ ~ ~ ~ ~ ~n . ~ .
V~ ~ ~ 4~ ''O ~ ~ ~ IA ~ V~ '.'~ ~ ~n . y, ' .
.... ~ ~-~
~ _ _ ..
H H H H H 1-1 H H 1:3 f~ 1 H H H H
~ ~ ~ ~-O O O O O O O O ~ X 1'- 0 0 C) O O ¢
O P )~- ~- 1' ~'-. rt ~ ~ O ~ I"' ~. ~.
r~ W ~ ~ ~ ~ o ~o O~ .J ~ ~ C O
~n o ~ o v~
Ul 3 0 ., O-~ ~ ~ ~'--H ~ ~1 C:~'. '~.... U~. .~O~D........ ,~J
_ _ _ ~ t~ ~?
v~ C _ ~ O
. ~ ' ~ C ~ ~ ..
~ o~ ", "~ n ~ n ~ ~ .
~ C~ - ~ W ~ ~
c.~ ~ ~
. . t~
n ~ _ . ~.. . ~
~ O ~ O ~ ~ 1~ ~J
- ' O ~ -J
C~ . . .
~ ,".~ ~' ` : :
mpl~. ~7 The residual ethanol c:ontent o~ the liposomQ
~u~p~nsions, produced according to exampl~ 2, b~or~ ~nd a.~tar ~reeze-dryinS~ i8 determined on 8 13atche~: by gas ahromat4~raphy. The ekh~nol content~ are det~rrnined in the original l~po~om~ ~uspen~ion with 0 . 5~ + 0 . 010% (m/V) .
x~lmPl~ 1~
Liposome ~u pen~ions and lyophilizate~ p~oduced according to example 2 ar~ stored at 4, 25 and 40~C. To ~udge th~ ~t~bility, app~aran~e, ~ize o~ in~ ion ~nd pH
-- in the c~ e o~ the lyophili2a~es a~ter resuspen~ion --~re determined at the re~pee~ve tim~.
A~tex 6evqral months o~ ~orage, the resuspended lyophilizate~ s}~owed no significant deviationæ relativ~3 to the ~x~mined ~3ize~ .
In pharma~:ological t~t~, the lipvsome susperla:ions thu~
produ-ed show th6~ ~ollowing properti~s:
~e~t ;~
Re uspended lyophilizates produced aGcording to example 2 ~re ~xarained relati~te to their aau~e toxic:ity after a ~ngle ~dmini~tration ~o a mou~ and rat.
'rh~ respeati~e LD50 is d~termined to b~:
.8 g o~ I/Rg body weight mou~e ~total iodin~) or . 0 ~ of I/kg body weight rat.
~5 ~
For re~uspended lyophiliz~'ces produs~ed aaaordin5~ to exampl~ ~, the ~ubac:u~ toxicity ~ ~: de'ce~min~d in rat~ ~n =
6) .
A~ter rep~ating the dv~e of l g of I/kg body weight ~total iodine) ~ time~, eaoh ~ an intsrval q~ 3 day~, no chang~ o~ hi~t~pathologioal or ~li~ic~l p~r~meter~ ~r~
de~rmined.
T~t ~
R~ u~pend~d lyophilizate~ ob~ain~d a~oording to ex~mple Z are examin~d rel~tive ~o ~heir organ connecti~n a~ter intravenou~ adminis~rat~on in rat~.
1 hour a~er th~ admin~ s~rati~n o~ 100 mg o~ ~/kg body weight (to~al iodine), 0.54 I~g o~ w~t weight corre~ponding to 24~ o~ the admini~ter~d do~ is ~etectad in th~ liver.
t Re~u pended lyophiliza~e~ obtained acaor~ o exampl~
2 ~re admini~tered intrav~nously in a do~ o~ lOO mg of total iodine/kg body weigh~ ~o rabbit~ With liYer tumors~
Th~ r~ulting differen~e in den~lty o~ ~iver/tumor i6 35 Houn~ied units (~U) after 1 hour.
:` Resu~pended lyophiliz~t~ o~ained a~cordlng to example ~0 2 are ex~min~d on do~s relative ~o their suita~ility ~or indirect C~ lymphography.
3 hours after in~er~igltal admini~tration ~$ 200 mg o~
total iodin~ ~in 3 ml of liposome suspension), at mo~t 200 ~U ~ density i~crea~e ~ 5 measured in ~ in the popllt~al ~5 and iliac lymph nodes.
~ e p~ceding examples C~ be repeated wit~ similar success b~ ~ubstituting the generic~lly or sp~ci~i~ally d~cribe~ reactant~ ~nd/or operating conditions o~ thi~
inv~ntion for tho~e us~d in the preceding axamples~
.,~ .
~ ~ L,- ~ ~t ~rom t;h~ ~oregoing de~orip~ion, one ~killed ~n the art can Rasily as~rtain ~he~ e sent~ al charac~rletics o~ thi~
inv~ntion, and withQut departir~g ~rom the spirit and sao~e thereo~, oan make variou~ changes and modiica~ion~ o~ the 5 inven~ion to adapt i~ to v~;rious u~ges and con~ition~.
phas~, high inclu ion capacitieæ or even a ~urther increase o~ ~he inclusion capacities ~an b~ achi~v~d.
In comparison wlth these previously known methods, the proces~ ~ccord~ng ~o the invention has ~he advant~g~ ~hat it i~ technically feaci~le on a large s~a~e in a simple wayA
Th~ u~ of strongly toxic or very ea~ily ~lammabls -~olvents i~ avoided. ~The ~ery ~ood reproducibility o~ tha pro~s~
according to the inven~ion i~ also ~o ~e empha~ized relative to the obtained a¢ti~e ingredient in~lu~ions ~nd li~osome -~izes, as well as the very hi~h active ingredient inclusion in the liposomes, up to over 50~, and the unu~ually high ~ctive in~redient ooncen~ra~ions, obtained ~y ~he proce~s accord~nq to ~he ~nven~ion, in the liposome 6usp~n~ion~ ~up to 800 mg/ml). Active ingredient inclusion means the percentage of the total active ingredient ~mployed which is contained in the inclusion ~olume of the lipids. ~ur~her, it is worth m~ntioning that it i~ possible to produae liposQme su~pensions, which have only a very l~w r~idual solvent cont~nt and which hav~ a good shelf li~e, with the help of the proc2s~ aocording to the invention. The proces~
ac~ording to the invention can al~o ~e per~ormed ~robl~m-fre~ under a~ptic conditions.
To perfor~ the proce~ according t~ th~ invention, ~he same substances for forming liposom~s can be used ~s in the previou~ly lcnown proce~-~es~
Sub~tanc~s ~orming ~uitable lipos4mes are u~ually amphiphatic lipid~ oX lipid mlx~ure~, ~uch a~, ~or ~xample, 2 ~ 7 ~
pho~pholipid~, sphin~omyel~nY or eth~r pho~phollpids. ~hese lipids aan h~ve straight-oh~ln or branched, ~aturat~d or un~aturated, similar or dif~erent acyl ~id~ chains.
Sult~ble phospholipld~ are, ~or example, the pho~phatldylchol~ne~, the phosphatidylethanolamine~, the phosphatidylserines, the pho~phatidylinosikol~, the pho~phat~dylglycerols or ~he pho~phatidia ~cids~ Sultable ether pho~pholipid3 are, for example, the plasmalogens (Dr. Otto~Albert Neumuell~r: Roempp~ C~2mi~-Lexikon;
Franak~che Verlagshandlung, Stu~tgart ~D~) 2665, 159, 3g20 and 4045).
For the production o~ liposom~s, lipids or mixtures thereo~ and in particular al~o mixtures of the~e ~ipid~ with cholesterol, chole~terol hemi~uooina~e, alpha-tvcoph~rol, alpha-tocopherol h~misuccinate and/or charg~ aarriers, such a~, for example, stearyl amine, ~teario ~aid, diethyl phosphate, oleic acid~ palmitic acid, bile acid, Cuch a~, for QXample, choli~ a~ld, glycocholi~ a~id, ~an ~e us~d.
Suitabl~ mixture~ can contain about up to ~0 mol~ parc~nt o~
cholester~l an~ up ~o ~0 mole peroent ~f charge c~rri~r. As solvent ~or the phospholipid3 or miX~ureR~ pre~erably isopropanol or in particular ethanol is ussd.
In principl~ the proa~ss aacording to the invention should al~o be feasible with water-~oluble low-boiling solvents o~her than lower a~cohols: for exampl~
t~trahydrofuran is suitable. But, performance of thi~
prooes~ embodiment i5 ~onsiderably ~ore expensiv~.
T~ perform the proce~ ~ccording to the invention, alcoholic ~olutt~n~ are pr~Qrably u~d which contain 0.
to 30 g of lipo~ome-forming ub~tance or su~ance mix~ure per lOO ml o~ ~olvent~ I~ nece6~ary~ the~e ~olutions are produ~ed by he~ti~g the components.
HPF~ i r~ .. lr~ I r~ _E! ,i~ l rEI_ 1 0-l:, i~ P0-~
~ ~ ,1, s Th~ proQass accerding to the invPn~ion i~ p~r~ormed, a~
wa~ already menti~ned, in such a way that the aqueous phase i~ added to ~he alcoholic solution with mlxin~ (such ~, for ex~mple, vigorou~ ~irrlng) and t~e lower alaoh~ re-moved, ~or exampl~, by vacuum di~tl llation or by introducingnitrogen. The mixing rat~ is g~ne~ally 50-1000 rpm~, pre-~e~a~ly 100-500 rp~.
Optlonally, th~ ~ispersion ob~ained after ~ombining the a~u~ou-~ pha~e and the al~ohollc 6ulution i~ mixed ~or so~e 10time ~about 10 to 1~0 minutes~ at a temperatura o~ about 20 to 90C, be~ore the solven~ is compl~tely or parti~lly sepa rate~. The volumstric ra~io of a~ueou~ phase ~o lipid pha~e is generally abol~ 50-1000 ml, pre~erably 100-500 ml o~
aqueou6 pha~e per lQ0 ml o~ lipid phase.
15The temp~ratur~s uit~ble ~or ~he proceY accordi~g to th~ invention are depend~nt on ~he solubility ~ th~ lipo-~ome-forming sub~tances ~ well a~ ~heir pha~e tran~itl~n temperature ~t~ he h~a~ stability of the activa ingr~-dient or a~tive ingr~dient mixtu~e ~nd ~h~ vapor pre~sure o~
~he alcohol to be ~epa~ated. ThP tempera~ure i6 pr~f~rably ~etween 20C and 90~C ~nd in partiaul~r b~tw~n 40~C and 70C. In general, it will be su~ ient to oper~te w~th a v~cuum of 10 h Pa to 100 h Pa.
The aqueou~ pha~e used ~or the pro¢es~ ac~ording to the in~ention can op~ionally ~ontain bu~er substan~e~ and/or i~otonizin~ add~ti~R~ Suitabl~ additl~es are, for example, inorganic or organiC ~alts or bu~fer subs~an¢e6, such ac 60dium chloride, ~ris buf~er, pho~pha~s buf~er, a~tr~t~
~uff er, glycine ~uf~r, citrate-pho~pha~e buffer, male~te bu~r, etc. Mono~ ~ di~ charides, ~uah as glu~ose, la~to~e, s~harose or trehalose, ~ugar alcohol~ ~u~h ~6 manni~ol, or~itol, xylltol or ylycerine or wate~-~oluble polymer~, ~uch as dextran or polyethlyene glycol.
Sinc~ th~ lipid~ and ~lso s~veral activ~ in~redi~n~s are se~itive to oxidation, th8 a~ue~u~ phas~ Gan be mixe~
with antioxldan~s, suoh s sodium a~cor~ate, tocophQ~ol or sodium bisul~ite.
The a~ueous l~posome susp~nsions produced aaco~d~ng to the proaess o~ the invention are used preferably ~or encap~ulaking water-~oluble actiVQ in~redi~nt3.
Suah water-soluble aa~ive ingr~dient~ are, ~or example, diagnostic agent~, such as the X-ray contr~t ~dl~
- lotrolan, iopromide, ioh~xol, iosimide, motriz~mide, s~lt~ of ami~oa~etic acid, iotroxic zcid, lop~midol, 5-hydroxyacetamido-2,4,6-triiodo-i~ophthalic acid-(2,3-dihydroxy-~-methylpropyl)-t~hydroxyethyl)-di~mide (=ZK
119095) and 3-carbamoyl-5-~N-(2~hydroxyethyl)-acetamido~-2,4,~-triiodo-benzo~ acid-~(lRS,2$~ ,3-dihydroXy-l~
hydro~ym~thylpropyl]-~mide (-ZX 13~129) or NM~ contra msdia, ~uch a~ gadolinium DTPA, yadolinium ~OTA and the gadolinium complex o~ 10-~1-hydroxymethyl-2,3-dihydroxypropy~ 4~7-~rls-t(carboxy~thy~ 4~7 tetraazacyclodecane~
Suitable ther~peutic aetive ingredients are, among others, antibiotio agents; such as gentamyoin or kanamycin, aytostatic ag~nt~, such a~ doxorubiain hydroohloride or cyclopho~pham~de and virustat~c ~ge~t~, such as ~idarabin~
or active ingredients, suah as mitoxantrone hydro~hloride~
These water-soluble active ingredi~nt~ ar~ dissolved in th~ a~ueous phase b~or~ per~o~mance o~ he proce~
according to the invention.
Furth~r, the ~queous liposoma susp~nsions aan ~l~o ~e US~d to encapsulata sli~h~ly soluble act~e ~ngr~dients in water.
Such active ingredient~ are, ~or example, plant protecting agents, such a~ ~lightl~ soluble inR~cticides or herbicid~s ~nd in ~ icular ~lightly oluble pharm~eutia~l active ingredient~.
,3~
Sligh~ly water-soluble or insoluble pharmaceutical active ~ n~redlents of the :~ollowing actlve ingredient group~3 are suitablet ~or example, for production o~ aqueous su~pen~;ion~ according to the proae~s cs~ the invention.
Ge~agenlcally a~:t$~e 6terc~id hormone~ uch a6, for ~xa~ple, 13-ekh~ 7~eta-hydroxy~ s-d1nor-l7alpha-pr~gn-4-en-20~ 3~ne ~levonorgestrel)~ 13-e~h~1-17be~a-hydroxy-18,19-dinor-17alpha-pregna-4,15-dien-20-yn-3-one (-ge~to~en) or 13-ethyl-17~e~a-hydroxy-11-methylene-18,1g-dinor-17alpha-1~ preqn-4-~n~20~yn (=de~oge~trel).
Estrogenally a~tive steroid hormones, such a~ 3-hydr~xy-1,3,5-(lo~-es~ra~rlen~17-on~ (-e~one~ or l,9-nor-17alpha-pregna-1,3,5~10~-trien-20-yn-3,17beta-diol ~thinyle tradiol).
Andro~ni~ally aati~ ~teroid hormone~, ~u~h a6, for example, 17~eta-hydroxy-4-andros~en-3-on~ (-t~sto~terone) and i~ e~ter or ~7b~ta-hydroxy-lalpha-methyl-5Alpha-androsten-3-on~ ~-me~terolone).
Antian~rogeni~ally ~c~i~e ~tero~d hormo~e~, such as, ~or example, 17alpha-aaetoxy-~-chloro-lbeta~2~eta-dihydro-3H-ayclopropa~l~2]-pregna-l~4~6-triene-3~2o-diono ~cypot~ronac~tat~)~
Corticoids, ~uch a~, for example, llbeta,17alpha,21-trihydroxy-4-pregnene-3,20-~ione (=hydroaortison~), llbe~a,17alpha,21-trihydroxy-1,4-pragnadi~ne-3,2~-dinne t=prednisolone), llbeta,17~1pha,21~trihydroxy~6alpha-methyl-1,4-pr~gnatriene-3,20-dione (-methylprednisolone) and 6~1pha-fluoro-llbeta,21 dihydroxy-l~alpha-methyl~1,4-prqgnadien2-3,2Q-dione (~ifluoortolone~ and their est~r~.
Ergolines, such as, ~or example, 3-~9,10-dihydro-6-methyl~8alpha-er~olinyl)-1,1-diethylure~ (=ergoline~
~romo-g,10 dihydro-6-mqthyl-8alpha-~rgolinyl~
3 ~
_ g ~
diethylurea ~=bromergoline) or 3-(6-methyl-8~1pha-ergolinyl)~ die~hylurea (=ter~ride).
Antihyper~e~sive agents, such as, for example, 7alpha-~cetylthio-17~1pha-hydroxy-3-oxo-4-pregnen~-21-carboxylic acid-gamma-laotone (aspironolactone) or 7alpha-ac~tylthio-15beta~1~bet~methylene-3-oxo-17alpha-pregna-1,4-diane-21,17-carbolactQns (~m~spirenone).
Antiaoagul~nt~, ~uch a~, f~r example, 5-[hexahydro-5-hydroxy-4-~3-hydroxy-4-methyl-1-octen-~-ynyl)-2(1H)~
1~ pentalenyliden~ pentanoi~ acid ~-iloprost~.
P~ychopharmacological ayents, suah a~, ~or exampl~, 4-(3-oyalopen~yl~xy-4-~ethQxy-phenyl-2-pyrrolidone ~rolipram) ând 7-chloro-1,3-dihydro-1-m~thyl-5-phenyl-2H~
1,4-bQnzodiazepin-2-one ~-diazepam).
Carotinoid~, auch a~, for example, ~lpha-oarotin and bata-carotin .
Fat-solubl~ vitamin~, ~uch a~, ~or example, vitamlns the vitamin A, vitamin ~, ~itamin E and vitamin K group.
Beta-oarbolines, as they ar~ de~cribad, ~or ~xample, in European patent appl~cation~ 234,173 ~nd 23~ 7, aro another group. A~ beta-carboline~, ~or ~xampl~, there can be mentloned 6~be~zoyloxy-4-m~thoxymethyl-b~ta-~rboline-3-carboxylic ~cid-isopropyl ester ~=b~carnil) and 5-~4-chloroph~oxy)-4-methoxymethyl-~eta-carboline~3~aar~0x~
a~id-1 opropyl est~r ~=Cl-P~O~IP).
Sligh~ly ~olubl~ or lnsoluble oon~ra~t m~dia are al80 worth mentioning, such a~ the X-ray contra~t medlu~
iodlpaml~e ethyl ester or NMR contrast media, such a~ the iron or manganes~ porphyrin chelates or ~l~o the ~gnetit~s.
As ~uitable active ingr~dients, salic~lic ~id, re~noic ~cld and azalaic acid further can be mentionqd~
Anti~y~otin~, such a~ the amphotericin B, ~conacol or meconazol ~an al~o be used.
-- 10 -- .
Before par~or~ance o~ the proce~3 aca6rding to the invention, th~:e aotive in~redient~3 ara disE;olved or su~:pended in the a}coholic solut:ion or ln the aqueous pha~e.
In the proces~ ~ccording to th~ invention, generally 0 . 05 to lo g and preferably 1 ~o 3 g o~ ac:tlve ingredient ls used per g o~ ~ubsta~nc~3 ~subs~anc6 mlxture) for forming liposom~. S~coril~ liposome suspensions can ~e produced in a ~i~nple way by the prooess ~coording to the inv~n~ion by both solutions being sterili7.ed by :Eiltration be~ore 10 per~ormanc~ o~ ~he proc~ss and all ~ubse~uent prooess 8~ep6 being per~ormed under a~eptic conditions.
~h~ inclu~ione~ which are achievable in the ll:posome ~u~pensions are dependent, o~ aourss, on tha typf3 o~ active in~redient and the substance ~ub tance mixture3 ~or ~orming 15 lipo om~a~a a~ w~ll a~ their ratio to one another.
Optionally, the unencap~ulated ~ctive ingredlen'c can ~e r~mo~rad, ~or examplo, by ul.trafiltxa~ion, dialy~
micro~iltration or ~entri~uging.
Tha lipo~ome ~uæpension obt~lned c:an be dl.rectly 20 stored. Alternatively, the llpo~ome su~pension can be converted by freeze-drying ~o a stable fo~m o~ ~orag~.
Be~ore the freeze- drying, if neoessary, among ~ther~, ~or example, additi~res, suah as mannitol or ~orbit~l and/or elec~rolytes and vis~osi~y-ln~luencing a~e~t~ su~h as sodium 25 chloride aan be added to the material~ to ~e fr~ez~-dried.
The lyophilizate~ can b~ re3usp~nded with bidistilled water or a~ueou~ phases, which ~an con~ain the same a~ditives aa th~ aqueouæ pha~e~ u~ed ~or lipo~ome prsduation.
Ths amount of r~suspQnsion m~dium can in this c~se b~
O.S t~ 20, but preferably 1 to 6 ml per g ~f lyophiliza~.
The ~u~pen~lon~ obtained can be uaed dlreatly or after ~iltration through filters of ~uitable pore ize.
2 ~ 7~
~ 11 ~
The lipo~ome~ produced by the proc~ss aecordlng to the ln~en~io~ ~.an, for example, b~ used i~ they contain ~ontr~3t media for N~R or X-ray dia~nosis in various diagno23~ic proc~s~s. ~hs~e include, for example, the di~gnosis of tumors in organs of the reticulo-endo~hellal ~ystem (~or example, liver and ~pleen~ or other i~trava3cul~rly acc~ible ti~3ue6 as well as the arthrography. The extrava~cular admini~ra~.lon o~ su~h lipo60~e ~usp~n~ion~
(such a~, ~or sxample, subcu~aneou~, intramuscular or intraperitoneal) ~an al~o be use~ ~or dia~no~is purpo3~s, ~uch as, for example, the indirect and ~irea~ lymphography, or for ther~pautl~ purpose~ ~such a~ intramu~cular depot prepara~lo~. Of c~ur~e, it i~ al~o po~sible to admin~ter tha preparations orally, and the prPparations opti~nally ~an be filled in aoat2d cap~ule~ rasist~nt ~o ga~tric jUiC~5.
Further, the liposom~ su~pensions containing active ingredients produaed ~y ~he pro~ aoco~ding to the invention can be applied in the ca~e o~ suita~le lipid compo~ition as blood pool ag~nt~ or a~ ~arget-directed pharm~c~utical vehicles.
Without further ~la~oration, it 1~ bel~eved ~hat one ~killed in the ~rt can, u~ing th~ pr~a~ding d~scription, utiliza the pr~sent invention o its ~ulle~t extent~ The ~ollowlng pre~erred speci~ic emhodiments are, ther~ore, ~o ~5 be construed a~ merely ~llu~tr~tlve, and not limitative o~
the remaind~r of the disclosure in any way whatsoever.
In the ~oregoing as~d in th~ ~ollowing ex~mples, al~
temperature~ are ~et ~o~th uncorrected in degree~ Celsiu~
and unle~s otherwiæe indioate~, all parts and peraentage3 are by weight.
The entir~ disclo3ure~ o~ all appli~ations, pat~nt~ ~d pu~lication~ ted ab~e and below~ and o~ corr~ponding 2 ~ 7 .j ~ 12 ~
appliaation Federal ~epublic o~ Germ~ny P 4~ 13 580.2, ~iled ~p~il 24, 19~0, are her~by incorporated by re~r~nce.
In the ~ollowing exampl~, th~ foll4wing ~breviation~
are u~ed:
PC - Ph~sphatidylcholine S 100 o~ the Lipoid KG
comp~ny CH = Powd~red ~hol~terol ~JP) ~ ~he Merck AG
company SA V~ry pure st~aric acid oP the Flu~a AG
company ~PPG = Dipalmitoylphosph~tidylylyaerol DPPG o~ th~
Lipoid KG company HSPC =~ Hy~rogenated 60yabean lQcithin ~Phospholipon 90H~ of the Na~t~r~ann AG company DCP ~ etyl phosphat~ of th~ ~igma company .
:
` `~
9.2~ g of lipid ~nixture (PC/~H/SA -- 4:5:1 mol:mc~ mol) is di~olved in loO ml ~ eth~nol and ~t~rllizad by ~iltrativrl at 70C. Th~n, th~ ~olu~iorl i~ trz~ns~rr~d to a reactiorl ve~6el he~ted moder~tely to 55~ and it i~ mixed with f3tirring (300 rp~ wi~h an ac~ive lngredient solution ~terlliæed by flltration (~ontaining ~7 . 75 g c~ ~ opromid~
~nd 200 ml o~ aqueou~ 20 mmol t:ri~ Cl b~fers pH 7~5~
lo T~en, th~ alcohol i~ ~listill~d of~ at 554C~ a vacuum ~I 5, 000 Pa and the re~ulting liposome su3pen~ion is axamined ~or its propertie3.
Then, the lipo~;ome su~pension i~ bottled dir~atly in portion~ of ~0 y in 50 ml gla~s in~usion bottl~ and it i8 ~r2~zQ-drisd. Th~ lyophillzates obtairled are re~usp~3nded 50 that an ~c:t~ve ingredient soncentration o~ a~out 2 oo mg/ml re~ults and al~o examined.
~L:L
Parformance o~ the te~;t talce~ place a; de3ar~ bed in ~0 exzlmpl~ xcept tha~ 18 . 5 g o~ iopromide is us~d.
Per~ormanc2 ot~ the ~ect takes pîac~ a: des¢ribe~ in exampls 2, except that 13 . 9 g of lipid mix1;ure i~ us~3d.
~xample 4 2~ P~arformancs of the te3t take~ placa as ds~cribed in ex~mple ~, except th~t 18 . 5 g af lipid mixture is u~ed~
-- 14 -~
~Ia S
P~rfor~anoe o~ ~h~ test took pl ace as desaribed in exam~pl Q 2, ~xcept that iotrol an i~; u~ed in~te~d of iopromide .
5 ~AP~ 5 Performance C)f the ~est takes placa a~; d~scri}:~ed in example 2, f3xc~pt that iopamidol is u~ed in ~cead o~
iopromide.
a~m~l~ .7 PerformanGe of tne te~t ta~es place as d~cribed in Rxample 2, eXc~pt that iohexal i~ used in~tead ~f i~promide.
~ca~pl ~3 ~
Per~o~ma~ce of ths t~st ~akes place a descri~ed in ex~mpl~ 2, ~xc~pt that the nonionic contra~t medium 5-hydroxyacstamide-~,4,~-tr~iod~ oph~h~lia acid-~,3-dihydrQ~y-N-methylpr~pyl~(2~hydroxy~thyl)-di~:ide ~ used in~tead of iopro~ide.
~x~mpl~ g Performa~ce of the ~e~t took pl~ce a~ desaribe~ in ~0 example 2, eXa~pt tha~ ~ lipid mixture of PC~C~/S~ 4:~
mol :mol :mol is used~
Per~ormanae o~ the taBt t~kes plac~ as des~ribed i~
example 2, except that a l~pid mixture of PC~C~/~C o~ 4:5:1 mol:mol:mol is u6ed.
J
~P~
Performance o~ the ~e~3t takes plaae ~ describ~d in exampl~ 2, exoept that a lipid mixture of PC/CH/SA o~ 6: 3 :1 mol: mol: mol i8 u~ed .
~ 2 Per~ormanc~ oi~ tha te~t take~ place a de~cri~ed in example 2, exc~pt that a lipid mixture~ o~ PC~C~/D~P of 4: 5 :1 i8 used.
Per~ormance o~ the te~t takes place as desc:ril:~e~ in example 2, exsept tha~ a lipid mixture o~ PC~C~I 1 s 1 is uaed .
Per~ormance o~ ~he te~ ~akes plaae as de2;crib~d in example 2, exc:ept that a lipid mixture of Pt::~HSPC/C~I/SA o~
2:2:5:1 i3 used.
Performance of the te~t take~ place as descri~ed in example 2, except that the batch size was ill~rea~d tenfold~
2 0 Per~ormanc:e o~ the test ~ake~ plac:e as descrihed in example ~, exc:ept that ~ one hunclred and ~ifty~old large~
batch 18 UBed.
~he followin~ table ~how~ the result~3 re~a~ed in examples 1 ~o 1~.
,, n, i7 ~.j . .
~;~X~
v ~ v ~ ~ ~ ~ ~ .
n ,. ~ <~ n ~ . g~ ~.
<~ ~ ~ ~ ~ ~ n ~ ~ ~ . ~ 1~
~ I ~ X ~ O 1 ~-V~ o ~ -o u~ ~ ~n ~ ~ ~ ~ ~ ~n . ~ .
V~ ~ ~ 4~ ''O ~ ~ ~ IA ~ V~ '.'~ ~ ~n . y, ' .
.... ~ ~-~
~ _ _ ..
H H H H H 1-1 H H 1:3 f~ 1 H H H H
~ ~ ~ ~-O O O O O O O O ~ X 1'- 0 0 C) O O ¢
O P )~- ~- 1' ~'-. rt ~ ~ O ~ I"' ~. ~.
r~ W ~ ~ ~ ~ o ~o O~ .J ~ ~ C O
~n o ~ o v~
Ul 3 0 ., O-~ ~ ~ ~'--H ~ ~1 C:~'. '~.... U~. .~O~D........ ,~J
_ _ _ ~ t~ ~?
v~ C _ ~ O
. ~ ' ~ C ~ ~ ..
~ o~ ", "~ n ~ n ~ ~ .
~ C~ - ~ W ~ ~
c.~ ~ ~
. . t~
n ~ _ . ~.. . ~
~ O ~ O ~ ~ 1~ ~J
- ' O ~ -J
C~ . . .
~ ,".~ ~' ` : :
mpl~. ~7 The residual ethanol c:ontent o~ the liposomQ
~u~p~nsions, produced according to exampl~ 2, b~or~ ~nd a.~tar ~reeze-dryinS~ i8 determined on 8 13atche~: by gas ahromat4~raphy. The ekh~nol content~ are det~rrnined in the original l~po~om~ ~uspen~ion with 0 . 5~ + 0 . 010% (m/V) .
x~lmPl~ 1~
Liposome ~u pen~ions and lyophilizate~ p~oduced according to example 2 ar~ stored at 4, 25 and 40~C. To ~udge th~ ~t~bility, app~aran~e, ~ize o~ in~ ion ~nd pH
-- in the c~ e o~ the lyophili2a~es a~ter resuspen~ion --~re determined at the re~pee~ve tim~.
A~tex 6evqral months o~ ~orage, the resuspended lyophilizate~ s}~owed no significant deviationæ relativ~3 to the ~x~mined ~3ize~ .
In pharma~:ological t~t~, the lipvsome susperla:ions thu~
produ-ed show th6~ ~ollowing properti~s:
~e~t ;~
Re uspended lyophilizates produced aGcording to example 2 ~re ~xarained relati~te to their aau~e toxic:ity after a ~ngle ~dmini~tration ~o a mou~ and rat.
'rh~ respeati~e LD50 is d~termined to b~:
.8 g o~ I/Rg body weight mou~e ~total iodin~) or . 0 ~ of I/kg body weight rat.
~5 ~
For re~uspended lyophiliz~'ces produs~ed aaaordin5~ to exampl~ ~, the ~ubac:u~ toxicity ~ ~: de'ce~min~d in rat~ ~n =
6) .
A~ter rep~ating the dv~e of l g of I/kg body weight ~total iodine) ~ time~, eaoh ~ an intsrval q~ 3 day~, no chang~ o~ hi~t~pathologioal or ~li~ic~l p~r~meter~ ~r~
de~rmined.
T~t ~
R~ u~pend~d lyophilizate~ ob~ain~d a~oording to ex~mple Z are examin~d rel~tive ~o ~heir organ connecti~n a~ter intravenou~ adminis~rat~on in rat~.
1 hour a~er th~ admin~ s~rati~n o~ 100 mg o~ ~/kg body weight (to~al iodine), 0.54 I~g o~ w~t weight corre~ponding to 24~ o~ the admini~ter~d do~ is ~etectad in th~ liver.
t Re~u pended lyophiliza~e~ obtained acaor~ o exampl~
2 ~re admini~tered intrav~nously in a do~ o~ lOO mg of total iodine/kg body weigh~ ~o rabbit~ With liYer tumors~
Th~ r~ulting differen~e in den~lty o~ ~iver/tumor i6 35 Houn~ied units (~U) after 1 hour.
:` Resu~pended lyophiliz~t~ o~ained a~cordlng to example ~0 2 are ex~min~d on do~s relative ~o their suita~ility ~or indirect C~ lymphography.
3 hours after in~er~igltal admini~tration ~$ 200 mg o~
total iodin~ ~in 3 ml of liposome suspension), at mo~t 200 ~U ~ density i~crea~e ~ 5 measured in ~ in the popllt~al ~5 and iliac lymph nodes.
~ e p~ceding examples C~ be repeated wit~ similar success b~ ~ubstituting the generic~lly or sp~ci~i~ally d~cribe~ reactant~ ~nd/or operating conditions o~ thi~
inv~ntion for tho~e us~d in the preceding axamples~
.,~ .
~ ~ L,- ~ ~t ~rom t;h~ ~oregoing de~orip~ion, one ~killed ~n the art can Rasily as~rtain ~he~ e sent~ al charac~rletics o~ thi~
inv~ntion, and withQut departir~g ~rom the spirit and sao~e thereo~, oan make variou~ changes and modiica~ion~ o~ the 5 inven~ion to adapt i~ to v~;rious u~ges and con~ition~.
Claims (15)
1. A process for the production of an aqueous liposome suspension comprising:
preparing a solution containing at least one liposome-forming substance in a lower alcohol solvent having 1-3 carbon atoms;
preparing an aqueous phase;
dissolving and/or suspending at least one active ingredient in said solution and/or said aqueous phase;
introducing said aqueous phase into said solution with mixing;
removing said lower alcohol by vacuum distillation or nitrogen introduction to form a liposome suspension; and optionally freeze-drying said liposome suspension.
preparing a solution containing at least one liposome-forming substance in a lower alcohol solvent having 1-3 carbon atoms;
preparing an aqueous phase;
dissolving and/or suspending at least one active ingredient in said solution and/or said aqueous phase;
introducing said aqueous phase into said solution with mixing;
removing said lower alcohol by vacuum distillation or nitrogen introduction to form a liposome suspension; and optionally freeze-drying said liposome suspension.
2. A process according to claim 1, wherein said active ingredient is water-soluble and is dissolved and/or suspended in said aqueous phase.
3. A process according to claim 1, wherein said active ingredient is a contrast medium.
4. A process according to claim 2, wherein said active ingredient is a contrast medium.
5. A process according to claim 1, wherein the active ingredient inclusion in the liposomes is up to 50%.
6. A process according to claim 1, wherein the active ingredient concentration in said liposome suspension is up to 800 mg/ml.
7. A process according to claim 1, wherein said solution contains 0.2-30 g of said liposome-forming substance per 100 ml of said lower alcohol solvent.
8. A process according to claim 1, wherein, after said aqueous phase is introduced into said solution and prior to removal of said lower alcohol, the combination of aqueous phase and solution are mixed for 10-180 minutes at a temperature of 20-90°C.
9. A process according to claim 1, wherein said vacuum distillation is performed at a vacuum of 10 h Pa to 100 h Pa.
10. A process according to claim 1, wherein said liposome suspension is freeze-dried to form lyophilizates and said lyophilizates are subsequently resuspended in an aqueous resuspension medium, the amount of said resuspension medium being 0.5-20 ml per g of lyophilizate.
11. A process according to claim 1, wherein 0.05-10 g of active ingredient per g of said liposome-forming substance is used to prepare said liposome suspension.
12. A process according to claim 3, wherein said contrast medium is an X-ray contrast medium.
13. A process according to claim 4, wherein said active ingredient is an X-ray contrast medium.
14. In a method for the diagnosis of tumors, the improvement wherein a suspension according to claim 1 is employed as a diagnostic medium and said active ingredient is a tumor diagnostic agent.
15. In a method of indirect lipography, the improvement wherein a suspension according to claim 1 is employed as a diagnostic medium and said active ingredient is a lipographic diagnostic agent.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DE4013580A DE4013580A1 (en) | 1990-04-24 | 1990-04-24 | METHOD FOR THE PRODUCTION OF ACTIVE AQUEOUS LIPOSOME SUSPENSIONS |
| DEP4013580.2 | 1990-04-24 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA2041075A1 true CA2041075A1 (en) | 1991-10-25 |
Family
ID=6405285
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA002041075A Abandoned CA2041075A1 (en) | 1990-04-24 | 1991-04-24 | Process for the production of aqueous liposome suspensions containing active ingredients |
Country Status (15)
| Country | Link |
|---|---|
| EP (1) | EP0478727B1 (en) |
| JP (1) | JPH04506814A (en) |
| AT (1) | ATE141503T1 (en) |
| AU (1) | AU651969B2 (en) |
| CA (1) | CA2041075A1 (en) |
| DE (2) | DE4013580A1 (en) |
| DK (1) | DK0478727T3 (en) |
| ES (1) | ES2093701T3 (en) |
| FI (1) | FI103178B (en) |
| HU (1) | HU215960B (en) |
| IE (1) | IE911350A1 (en) |
| NO (1) | NO303481B1 (en) |
| PT (1) | PT97468B (en) |
| WO (1) | WO1991016039A1 (en) |
| ZA (1) | ZA913091B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7901708B2 (en) | 2002-06-28 | 2011-03-08 | Protiva Biotherapeutics, Inc. | Liposomal apparatus and manufacturing methods |
| US9005654B2 (en) | 2005-07-27 | 2015-04-14 | Protiva Biotherapeutics, Inc. | Systems and methods for manufacturing liposomes |
Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0660714B1 (en) * | 1991-06-18 | 2003-07-02 | ImaRx Pharmaceutical Corp. | Novel liposomal drug delivery systems |
| WO1995004523A1 (en) * | 1993-08-06 | 1995-02-16 | Opperbas Holding B.V. | A method for preparation of vesicles loaded with biological structures, biopolymers and/or oligomers |
| ATE219688T1 (en) * | 1994-03-28 | 2002-07-15 | Nycomed Imaging As | LIPOSOMES CONTAINING AN X-RAY OR ULTRASONIC CONTRAST AGENT |
| WO1997035559A2 (en) * | 1996-03-27 | 1997-10-02 | Ortho Pharmaceutical Corporation | Manufacture of liposomes and lipid-protein complexes by ethanolic injection and thin film evaporation |
| WO2000045791A2 (en) * | 1999-02-08 | 2000-08-10 | Alza Corporation | Method for controlling liposome size |
| CN114344291B (en) * | 2022-02-16 | 2023-09-29 | 中南大学湘雅医院 | Application of 4-phenyl-1H-pyrrole-3-carboxylic acid in preparation of medicine for treating osteoporosis and liposome |
Family Cites Families (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE3416978A1 (en) * | 1983-05-12 | 1984-12-06 | Stauffer Chemical Co., Westport, Conn. | TRANSFORMATION OF EUKARYOTIC CELLS MEDIATED BY LIPOSOME |
| JPS607932A (en) * | 1983-06-29 | 1985-01-16 | Dai Ichi Seiyaku Co Ltd | Preparation of liposome |
| EP0158441B2 (en) * | 1984-03-08 | 2001-04-04 | Phares Pharmaceutical Research N.V. | Liposome-forming composition |
| FR2634375B3 (en) * | 1988-06-30 | 1991-07-05 | Centre Nat Rech Scient | PROCESS FOR THE PREPARATION OF DISPERSIBLE COLLOIDAL LIPID AMPHIPHILIC SYSTEMS IN THE FORM OF SUBMICRON LIPOSOMES |
| JP2554880B2 (en) * | 1987-05-07 | 1996-11-20 | ポーラ化成工業株式会社 | Skin cosmetics |
-
1990
- 1990-04-24 DE DE4013580A patent/DE4013580A1/en not_active Withdrawn
-
1991
- 1991-04-04 AT AT91906756T patent/ATE141503T1/en not_active IP Right Cessation
- 1991-04-04 HU HU9200235A patent/HU215960B/en not_active IP Right Cessation
- 1991-04-04 EP EP91906756A patent/EP0478727B1/en not_active Expired - Lifetime
- 1991-04-04 ES ES91906756T patent/ES2093701T3/en not_active Expired - Lifetime
- 1991-04-04 WO PCT/DE1991/000294 patent/WO1991016039A1/en active IP Right Grant
- 1991-04-04 DK DK91906756.1T patent/DK0478727T3/da active
- 1991-04-04 DE DE59108097T patent/DE59108097D1/en not_active Expired - Fee Related
- 1991-04-04 JP JP3506736A patent/JPH04506814A/en active Pending
- 1991-04-23 IE IE135091A patent/IE911350A1/en unknown
- 1991-04-24 CA CA002041075A patent/CA2041075A1/en not_active Abandoned
- 1991-04-24 PT PT97468A patent/PT97468B/en not_active IP Right Cessation
- 1991-04-24 ZA ZA913091A patent/ZA913091B/en unknown
- 1991-04-24 AU AU75933/91A patent/AU651969B2/en not_active Ceased
- 1991-12-18 NO NO915012A patent/NO303481B1/en unknown
- 1991-12-23 FI FI916120A patent/FI103178B/en active
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7901708B2 (en) | 2002-06-28 | 2011-03-08 | Protiva Biotherapeutics, Inc. | Liposomal apparatus and manufacturing methods |
| US8329070B2 (en) | 2002-06-28 | 2012-12-11 | Protiva Biotherapeutics, Inc. | Liposomal apparatus and manufacturing method |
| US9492386B2 (en) | 2002-06-28 | 2016-11-15 | Protiva Biotherapeutics, Inc. | Liposomal apparatus and manufacturing methods |
| US9504651B2 (en) | 2002-06-28 | 2016-11-29 | Protiva Biotherapeutics, Inc. | Lipid compositions for nucleic acid delivery |
| US11298320B2 (en) | 2002-06-28 | 2022-04-12 | Arbutus Biopharma Corporation | Liposomal apparatus and manufacturing methods |
| US11318098B2 (en) | 2002-06-28 | 2022-05-03 | Arbutus Biopharma Corporation | Liposomal apparatus and manufacturing methods |
| US9005654B2 (en) | 2005-07-27 | 2015-04-14 | Protiva Biotherapeutics, Inc. | Systems and methods for manufacturing liposomes |
Also Published As
| Publication number | Publication date |
|---|---|
| FI103178B1 (en) | 1999-05-14 |
| AU651969B2 (en) | 1994-08-11 |
| HUT62457A (en) | 1993-05-28 |
| ZA913091B (en) | 1992-12-30 |
| IE911350A1 (en) | 1991-11-06 |
| EP0478727A1 (en) | 1992-04-08 |
| EP0478727B1 (en) | 1996-08-21 |
| FI103178B (en) | 1999-05-14 |
| DK0478727T3 (en) | 1997-01-20 |
| NO915012L (en) | 1991-12-18 |
| FI916120A0 (en) | 1991-12-23 |
| DE4013580A1 (en) | 1991-10-31 |
| PT97468A (en) | 1992-01-31 |
| AU7593391A (en) | 1991-11-07 |
| ATE141503T1 (en) | 1996-09-15 |
| WO1991016039A1 (en) | 1991-10-31 |
| HU215960B (en) | 1999-03-29 |
| HU9200235D0 (en) | 1992-04-28 |
| DE59108097D1 (en) | 1996-09-26 |
| JPH04506814A (en) | 1992-11-26 |
| PT97468B (en) | 1999-12-31 |
| NO303481B1 (en) | 1998-07-20 |
| ES2093701T3 (en) | 1997-01-01 |
| NO915012D0 (en) | 1991-12-18 |
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