CA1320400C - Method and apparatus for producing lipid vesicles - Google Patents
Method and apparatus for producing lipid vesiclesInfo
- Publication number
- CA1320400C CA1320400C CA000592763A CA592763A CA1320400C CA 1320400 C CA1320400 C CA 1320400C CA 000592763 A CA000592763 A CA 000592763A CA 592763 A CA592763 A CA 592763A CA 1320400 C CA1320400 C CA 1320400C
- Authority
- CA
- Canada
- Prior art keywords
- phase
- inlet
- lipophilic
- mixing chamber
- orifice
- 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.)
- Expired - Lifetime
Links
Classifications
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B65—CONVEYING; PACKING; STORING; HANDLING THIN OR FILAMENTARY MATERIAL
- B65B—MACHINES, APPARATUS OR DEVICES FOR, OR METHODS OF, PACKAGING ARTICLES OR MATERIALS; UNPACKING
- B65B1/00—Packaging fluent solid material, e.g. powders, granular or loose fibrous material, loose masses of small articles, in individual containers or receptacles, e.g. bags, sacks, boxes, cartons, cans, or jars
-
- 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
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
- B01F25/00—Flow mixers; Mixers for falling materials, e.g. solid particles
- B01F25/10—Mixing by creating a vortex flow, e.g. by tangential introduction of flow components
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J13/00—Colloid chemistry, e.g. the production of colloidal materials or their solutions, not otherwise provided for; Making microcapsules or microballoons
- B01J13/02—Making microcapsules or microballoons
Abstract
ABSTRACT
The present invention provides an apparatus and method for manufacture of multilamllar or paucilamellar lipid vesicles. The apparatus and method use shear mixing in a substantially cylindrical mixing chamber having at least one tangential input for rapid production of the lipid vesicles.
The present invention provides an apparatus and method for manufacture of multilamllar or paucilamellar lipid vesicles. The apparatus and method use shear mixing in a substantially cylindrical mixing chamber having at least one tangential input for rapid production of the lipid vesicles.
Description
METHQD AND APPARATUS FOR PRODUCIN~
I~E~
1 Backqroun~ of the In~ention The present ln~ention relates to a metho~
an~ apparatus for producing lipi~ vesicles More particularly, the invention ls ~irecte~ to a S pro~uctlon technique that permit~ rapi~, high volume formatlon of llpi~ ve~lcle~ from ~urf~ctants n~
other amphlphlllc molecule~.
~lpl~ ve~lcle~ aro llpl~ ~tructures whlch ~urrouna ~n~ encapsulat~ a~ueoua volume~ Thero are lo ma~y u~e~ for these ~tructures, ~ g , as ~uva~t~ or ~8 carr~ers for tbe transportatlo~ of encapsulate~
I~E~
1 Backqroun~ of the In~ention The present ln~ention relates to a metho~
an~ apparatus for producing lipi~ vesicles More particularly, the invention ls ~irecte~ to a S pro~uctlon technique that permit~ rapi~, high volume formatlon of llpi~ ve~lcle~ from ~urf~ctants n~
other amphlphlllc molecule~.
~lpl~ ve~lcle~ aro llpl~ ~tructures whlch ~urrouna ~n~ encapsulat~ a~ueoua volume~ Thero are lo ma~y u~e~ for these ~tructures, ~ g , as ~uva~t~ or ~8 carr~ers for tbe transportatlo~ of encapsulate~
drugs or biologically-active substances. Lipi~
vesicles are often classifie~ into three groups by size an~ structure: multilamellar vesicles (~MLV'5~), large unilamellar vesicles ~L W 's~), and small unilamellar vesicles (~S W 's'). ~LV'8 are onion-like structures having a serie~ of substanti~lly spherical shells forme~ of lipid b~layer~ intersperse~ with aqueous layers. L W 's have a ~iameter greater than 1 ~ ana are forme~ of a lipi~ bil~yer ~urroun~ing a 10 large, unstructure~ a~ueous phase. S W 'fi are ~imilar in ~tructure to the L W '~ escept their ~ameters are less than 0.2 p.
A ourth type of lipi~ vesicle, which is partioularly well suite~ for transport of either lipiDs or aqueou~ materials, i~ the paucilamellar vesicle (~PLV~ Q, Callow and McGrath, Cryobiology 1985, ~3), pp. 251-267. This type of vesicle has an e~ternal structure of about two tc five peripheral lipid bilayers with a large, 20 unstructure~ aqueous center. Lipid droplets, e.g., oil droplets, may be suspended in the aqueous center, lea~ing to very high uptake of aqueous or lipophilic materials. Paucilamellar vesicles range from about 2-15 ~ in ~iameter.
E~ch type of l~pi~ vesicle has distin~t advantages for certa~n uses. Because of the relatively large amount of lipi~ ln the lip~
bilayer~ of the NLV's, they are consi~ere~ best for encapsulation or transportation of l~pophille 30 material~. She L W '~, because of the~r large aqueous~lip~ volume rat~o, are consi~ere~ best for .~;, encapsulation of hydrophilic molecules, particularly macromolecules. S W 's have the advantage of small size which allows relatively easy access to the cells of tissue, but their small volume limits delivery o~
hydrophilic aqueous materials to trace amounts.
However, S W 's may be useful in place o MLY'~ for the transportation of small guant~ties of lipophilic material~ because of high lipid~water ratios. PLV's can tran~port largs quantities of agueous or lipophilic material~ but their large size can preclude approach to certain tissues.
The pre~ent invention pertains to the formation of MLV's an~ PLV's. Since SW '~ are commonly made by sonification of multilamellar lipid Yesicles, it follows that the processes by which MLV'~ are produced can be used as part of a technigue for making SW'8.
The conventional approach to producing multilamellar lipid vesicles, particularly liposomes maDe of pho~pholipi~s, ~tarts by difisol~ing the lipid~, together with any lipophilic aaditlves, in an organic solvent. The organic solYent is then removed by evaporation using heat or by passing a stream of an inert gas (e.g., nitrogen) oYer the dissol~e~
lipids. ~he resi~u~ ~s then hydrated with an agueous phase, generally containing electrolytes an~
a~ditives ~uch as hyaroph~lic biologlcally-actiYe materials, to form large multilamellar llpl~ membrane structuro~. In rome ~ariat~ons, ~ifferent types of 30 partlculate mattor or stru~tures have been u~e~
~uring the ovaporation process to assist in the ~ ~3~0400 formation of the lipi~ residue. Those in tbe fiel~
have shown that by changing the physical structure of the lipid resiaue, better vesicles form upon hydration. Two recent review publications, Szoka ana Papahdjopoulo~, Ann. Rev. Biophy6. Bioeng. 2:467-508 (1980), an~ Dousset ana Douste-Blazy ~in LL~sssme~, Puisieus ana Delattre, E~itors, technigue~
et Documentation Lavoi6ier, P-ris, pp.~l-73 ~1985)), summarize the methoa6 which have been use~ to make MLV ' .
~o matter how the MIV'~ or PLV'~ are formea, once maae it i8 necessary to determine th~
effectiveness of the process. ~wo measurement~
commonly usea to determine the effectiveness of encapsulation of materials in lipi~ vesicles are the encapsulatet mass and capturea volume. The encapsulated m~s is the masfi of the substanco encapsulatea per unit mass of the lipia ana ~s often given as a percentage. The capture~ volume i~
defined as the amount of the aqueous phase trappe~
inside the vesicle aiviaea by the amount of lipi~ in the vesi~le structure, normally given in ml ligui~g lipi~.
A ai6a~vantage as~ociatea with proaucing MUV'~ or PLV'~ usinq stan~ard metho~s ~s that these processes are costly, slow an~ relatively ineffic~ent ~n terms of matorial. For eYample, the stan~ard time to manufacture M~Y's ~ ~n the or~er 2-20 hour~. If ~ W '~ ~re rogulre~, the sonication which break~ the multila~ellar lipi~ ~tructures ~nto 8 W '- take~
a~itlonal t~mo. I~iJ ~low processing i~ unwiel~y s 1320~0o - -an~ e~pensive for any large scale production of lipi~
ve~icles .
While rapi~, continuous-flow, mi~ing processes are known in other arts, e.g., chemical kinetics, tbe a~aptation of such processes for the production of lipiD ~esicles has not heretofore been deviseD, or even suggesteD.
Accordingly, an ob~ect of the invention i~
to provi~e an improvea metho~ and apparatus for making multilamellar or paucilamellar lipid vesicle5.
A further object of the invention is to provide a lipid vesicle forming technique which len~s itself to commercial, high-volume production.
Another object of the invention to provide a metho~ and apparatus for the rapid, efficient encapsulat~on o biologically-active macromolecules into lipid vesicles.
These an~ other object~ an~ features of the in~ention will be apparent from the following 20 descriptlon an~ firawings.
~ummar~ of the ~nvention She presen~ invention feature~ a metho~ of pro~ucing multilamellar or paucilamellar lipi~
vesi~le~ and an apparatu8 useful ln thelr proDuction. She metho~ i~ base~ on ths shear mi~ing of a ~olventless lipophilic phase, an~ a hy~rophilic or aqueoug phase ~n order to rapi~ly hy~rate the lipophilic phase, thereby formlng multilamellar or paucilamellar ~tructures. ~he ~ifference in structure is dependent, ln part, on the choice of materials forming tho lipophilic pha~o.
The method of the in~ention uses a mising chamber having a substantially cylin~rical lnner surface with at least ono substantially tangentially locate~ ~nlet orifice which allow flow of one phase, 10 preferably tho lipi~ phase, into the hollow interior. An ad~itional inlet orifice, which may be tangentially or ~ubstantially asially locate~, provi~es input of the other phase, preferably the aqueous phaso. There i8 also a substantially asially locate~ outlet orifice which allows removal of the l~pi~ vesicles after formation.
Th~ l~pophilic phase i~ forme~ of -surfactant, preferably blen~e~ with ~ steroi~ such a5 cholesterol an~ an amphiphilic charge-pro~uc~ng 20 material, a~ well a8 any lipophilic material~ to be incorporated into the lipi~ vesiclo. Thi~ lipophilic phase iB injecte~ into the mising chamber through at least one of the inlet or~fices. An agueous phase, which contain~ primarily water or buffer an~ any hy~roph~l~c material~ to be incorporate~ into tho l~p~ ve~iclo, enter~ the ~ising chsmber through a ~ubstantially tangentially locats~ input orif~ce or an asially locate~ inlet orifico. Both tho l~poph~lic pha~ an~ agueou~ pha~e ar~ in~ecto~ with 30 ~ufficient force to croat- a rap~ ubstantially flo~, ~rsferably tangont~al, about the inner ~urface ~7~ 13 2 0 ~00 of the mi~ing chsmber. The two streams of flowing liquid, the lipophilic phase and the aqueous phase, intersect in such a manner as to cause shear mi~ing, thereby forming the lipid vesicles. Turbulent flow for each liguid i8 preferred but may be unnecessary so long as the two phases intersect with sufficient force. The formed lipid vesicles are removed from the chamber through the asial outlet orifice and collected. In the preferred embodiment of th-lnvention, two tangential flow~ of liquid intersect when travellng in the same direction, e.9., both clockwise or counter-clockwise about the inner surface of the mis~ng vessel. Depending on the pump speed, this flow allows shear mi~ing without causing e~cessive turbulence which would slow down the net flow. ~Shear mising~ is define~ as the mising of the lipophilic phase with the agueous phase un~er turbulent or ~hear con~itions which provide a~eguate mising to hyarate the lipid and form lipid vesicles.
20 The pump speeds are modified depending on the viscosity of the materials and the size of the orifices selected. ~Shear mi~ing~ is achieved by liqui~ shear which i~ substantially eguivalent to a flow rate for the combined phases of 5-30 m~s through a 1 mm radius oriflce. In Dnother embodiment of the invention, the two ~treams of fluid intersect by traveling in countercurrent directions, e.g., one clockwlse and one counter-clockw~e. Thl~ 18 most u~eful fo~ very dilute ~olutions where escea~i~e 30 turbulence and resultin~ back pressure is not as much of a problem.
-8- 1~ 20~100 Preferred surfactants for use ~n the ~nvention lnclude polyosyethylene acyl ether~, polyo~yethylene acyl amine~, polyosyethylene alkyl esters, polyosyethylene ~orbita~ alkyl esters, and polyosyethylene glycerol alkyl ester~, where sa~d ~
alkyl or acyl groups have 16-20 carbon atoms and not more than two ~ouble bon~g. Many of these materials are liquid at room temperature ~o no heatlng or other treatment ~tep is necessary to achieve the fluld 10 stage helpful for rap~ flow. If the mater1als are not liguid at operating temperature, heating befor~
introduction through the tangential inlet orifice ls a preferre~ step since the liqul~ state provi~es proper flow about the mising chamber.
Selection of the proper surfactant can modify the structura of the final lipid vesicles.
Certain surfactants will form PLV's wh~le others will form MLV's. Although not necessary to the invention, the difference 1~ theorized to occur because of the 20 ~ifferences in the lipophilic~hydrophilic balance of the surfactants.
The chamber should be ma~e of materials whlch can withstand ~olut~ons whlch are heated above the melting point of the lipi~. A chamber which will withstand ~0C. ~a temperature high enough to melt ~ubstantially all of the llpids within thls class) ~s helpful.
Th~ inv-ntlon al80 feature~ a ~evlce or ma~lng llp1d v-~icles. Th~ central part of thls 30 de~ice 1s the mlsing chamber w~th the inputs as -9_ 1320400 previously descri~ed. In addition, this device should have reservoirs for the lipophilic and hydrophilic phases an~ pumps or drive mechanisms which force the phases, preferably in fluid form, - through the inlet orifices into the mising chamber.
In addition, the inlet path from the reservoirs, or optionally the reservoirs themselves, can include mising device insuring uniform dispersions of each of the lipophilic an~ ~gueous phases. Another option i8 to have heaters, either withln the reserYoir or between the reser~oir and the mising chamber, to allow heating of the phases if necessary for proper liguid flow.
The ~pecifics of the pumps, feea lines, and orifice ~ize selection are a matter of engineering within the skill of those practicing the art and depend on amount an~ viscosity of the material to be processed and the desired flow rates.
These an~ other features of the invention 20 will be more apparent from the drawings and the det~iled description.
Brief Description of the Drawins~
FI¢URE 1 i~ an perspect~ve view of 8 mising chamber for use ~n accordance with the pr~ctice of the ~nvent~on;
FIGURE 2 ~ a ~ectional v~ew taken along l~ne I-I of the mising chamber of FIGURE l;
~o 1320~00 FIGURE 3 i~ a schematic representation of a lipi~ vesicle producing apparatus in accordance with a first embodiment of the invention; an~
FIGURE ~ i8 a schematic representation of a lipi~ vesicle pro~ucing apparatus in accordance with a ~econ~ embo~iment of the invention.
Detaile~ Description of Preferre~ Embo~iment~
As note~ previously, the present invention features a metho~ of making multilsmellar or lO paucilamellar l~pi~ ~e~icle~ an~ a ~evice for the manufacture of those lipid vesicles. The metho~s an~
~evice have a mising chamber with a substantially cylin~rical interior Qurface as a central feature.
This mi~ing chsmber permits input of material~ with controlled intersection of the lipophilic an~
hy~rophil~c phase~ ~n or~er to form the multilamellar or paucilamellar lipi~ ve~icles. ~he tsrm ~substantially cylin~rical~ as use~ herein means an~
~nclu~es cylin~r~cal, frustoconical an~ other ~hapes 20 having substantially similar surfaces of re~olution.
The following description will more clearly eluci~ate the principleR of the invention.
l. The M~ina Chamber A mising chamber ~uitable $n the pre~ent ~n~entlon ~ ~hown ln FIGUXE l. Mi~ng chamber 20 h~ a ~ub~tant~-lly cyl~n~rical cro~-section form~ng ~ central, hollowe~ out portion of a block of -ll- 1320~00 material 300. Inlet feed lines, illustrateD as 310A-310D, allow the lipophilic anD agueous phases to be in~ecte~ into mising chamber 20. Each of these inlet feed lines 310A-310D ends in an inlet orifice 320A-320D.
FIGURE 2 18 a cross-sectional illustration of mising chamber 20 about lin~ I-I. As shown in FIGURES 1 anD 2, mi~ing chamber 20 has a cylin~rical inner ~urfac~ 330 DisposeD about a central asis ~Desi9nateD ~c~) anD i8 close~ at each enD by a circular face 332 anD 334. Discharge orifice 350 ~s disposeD through one of the circular faces, for esample, as shown, through face 332. Discharge orifice 350, which preferably has of circular cross-section centereD on the central asis c, ha~ a ~iameter ~ubstantially less than that of the cylinDrical surface 330. In this preferre~
embodiment, discharge orifice 350 i~ both asially and raDially spaced from the inlet orifices 320A-320D.
Inlet orifices 320A-320D ser~e as in~ection ~ets anD are arrangea in paceD relation perpherally about mising chamber 20. While it is preferreD that inlet orifices 320A-320D, as well as inlet fee~ lines 310A-310D, are arrangeD in a plane ~ispo~eD
perpenD~cular to the central asi~ c of the cylinDrical mlsing chamber 20, th~ ~s not requireD.
The four ~nlet orifices 320A-320D shown ~n FIGURE 1 are ~paeeD about the cy~in~rical inner ~urface 330 at nin~ty ~egres ~ntor~al~ anD are tangential relati~e 30 to cylinDrical innor ~urace 330.
-12- 13~0~00 Lipi~ material is introduced into mising chamber 20 through inlet orifices 320A and 320B which are connectea to esternal feed lines 158 and 162 (F~GURE 3) through inlet feed lines 310A-3108 ~t diametrically oppo~ite si~es of cylindrical ~urfsce 330. The agueous phase i8 introducea through inlet orifices 320C an~ 320D which are connecte~ to esternal feed lines lC0 and 164 ~FIGU~E 3) through inlet feed lines 310C an~ 310D at di~metrically opposite ~ides of cylindrical ~urface 330. With the illustrated arrangement, the fluids flow rotationally, as indicated by ~rrow ~a~, within the mising chamber 20. All the fluids flow in the ~ame direction, either clockwi~e or counter-clockwise, depending on whether viewed from face 332 or face 334.
The flow rate~, volumes and particularly the flow paths are key~to the efficient production of quality vesicles according to the invention. The fluids enter the mi~ing chamber 30 tanqentially with respect to cylindrical wall 330, ~pprosimately 90-from the other inlet orifices. The flui~ stream 1 guide~ ~n ~ ~piral flow path from each ~n~ect~on orifiee 310A-310D arouna the inner ~urface of the cylindrical w~ll 330, ~nd then out through d~scharge oriflco 350 ~nto an oslt tube 170. $he flow p~th~
are controlled by the or~ent~tion ~nd pl~cement of the ~n~ection orifices 310A-310D 80 a8 to intersect, creating the mi-~ng rone. The pump ~pee~, a~ well as the orifice an~ fee~ l~ne ~iameters, are ~electea 30 to achieve proper ~hear mising for l~pi~ vesicle format~on. In mo~t c~rcumstance~, turbulent flow rolecte~ to prov~e ~equ~to mis~ng.
2. Pump Dr~ven Flui~ DeviGg As note~ previously, the mising chamber is the heart of the apparatus for making the multilamellar or paucilamellar lipi~ vesicle~.
FIGURE 3 illustrates one embodiment of the invent~on which uses th~ mising chamber.
~ i~ing chamber 20 (not shown) i~ within block 300. She lipid reservoir 40 hol~s the lipophilic pha~e an~ the agueous reservoir S0 hol~s 10 the agueous phase. Connected to the reservoir~ an~
the outlet from the mising chamber are the first, second and third pump~ 60, 62 and 64 with their respective controller~ 70, 72 an~ 74. Thir~ pump 64 and it~ controller 74 are optional and are unnece~sary un~er most circumstance~.
The lipophilic phase, consisting the surfactant an~ the other lipophilic materials ~e.g., a steroid such a~ chole~terol, a charge-pro~ucinq amphiphile, an~ any lipophilic materials to be 20 encapsulated) are blended and placed in lipi~
reservoir 40. Heater 30 ~ optional an~ i8 meant to illustrate any device which can preheat the lipophilic phass (if neede~) to allow it to flow a~ a flui~ at tempersture of operation. Heater 30 may be a hot plate, an oven, or the entlre apparatus may be place~ in a warm room.
Preferably, the temperature of the lip~
material within hol~ing reservoir 40 i8 such that the lipi~ material remain~ ~n a flui~ ~tate. For lipi~
-l~- 1320400 materials with melting points above room temperature, ~t maybe necessary to keep the material at elevate~ -temperature within the reservoir. So that en~, holding reservoir 40 can serve as an incubator, optionally having a warming element such as tho illustrate~ heater coll 106 which is helically wound about the interior holding ves~el 102, and~or -thermally insulating ~ac~et enclosing hol~ing vess-l 102. ~n certain instance~, it 1~ ~esirable to melt the lipid material directly within holding reservolr 40 by employing warming element 106, thereby eliminating the nee~ for separate heating ~evice 30.
If the lipi~ is a ligui~ at room temperature, no heating device i~ necessary.
- Lipophilic additives, e.g., steroi~s such as cholesterol, are normally blended into the lipid before ths lipophilic phase reaches holding reservoir 40. However, additives can be introduced ~irectly into holding reservoir 40 and blendea into ths 20 lipid. If 80, a mising device such 8S a magnetic or mechanical miser 108 is provide~. Illustrated miser 108 has a vaned, mising element 110 e~ten~ing longitu~inally within the vessel 102. Mising element 110 is rotationally driven by a driver, such a~ a motor or an electromagnetic driving coil 112 disposed outside holding ves~el 102, with which mising element 110 i~ magnetically couple~.
Lipi~ re~ervoir 40 is coupled to the inlet of pump 60 by tubing 132. Preferably, pump 60 i8 a 30 po~itive ~l~placement, magnetically ~riven pump. lt ~ al~o preferred that the pump be detachable from -15- 1~20400 ~t~ dr~ving motor~ (not ~eparately ~hown) for steril~ing, for esample, by autoclav~ng. A ~u~table pump for the l~p~ mater~al, for e~ample, ~ made by Cole-Parmer, Model ~o. J-7002-23, coupled with motor drive Model No. 7144-0~.
The output from pump C0 ~ connected to ml~lng chamber 20 by fee~ line 150. Feed l~ne 150 ~8 provided wlth a flow ~plitting or branch f~ttlng arran~ement lC6 whereby fir~t llne lS~ 1~ connect~ad to both estern~l feed llne 158 and esternal feed llne 162. Feed line 150 ha~, for esample, a 1~4 lnch (0.1 cm) outer dlameter and a 1~8 lnch ~0.05 cm) inner dlameter. All connectlons are pre~era~ly made with compression fitting~, for esample, of Teflon brand material which can be autoclaved at the ~ame time ~s the lines.
Each of the esternal feed llnes 158 ~n~ 162 are connecte~ to inlet fee~ llnes for the l~poph~lic phase, specific~lly inlet feed lines 310A and 310B, 20 respectively.
FIGURE 3 al~o illustrates the agueous phase portion of the system. Aqueous reservoir S0 ~
~imilar ln construction to llpi~ re~ervolr ~0 escept cham~er 122 ~ 8 larger ~ecauso the aqueou~ volume ~dde~ to hydrate the lipophilic pha~e i~ normally much larger than the llpi~ volu~e. Neater 126 ~
optional an~ ia used normally if the agueou~ phase ~s heate~. Thi~ permlt~ the reaction to ta~e pl~ce witbout largc temperature aifference~ which provi~e~
30 better lipl~ vesicl~ formatlon.
*Trade Mark -16- 1320~00 The aqueoua ph~se, ~nclud~ng any a~t~vea or b~ologicals which are to be encapsulated, ~
place~ In agueou~ reservo~r 50 an~ flow~ through tubing 13~ to pump 62. Aga~n, pump 62 Ig preferably a positlve d~splacement, magnetieally drl~en pump w~th a detachable ~rl~e motor. A Cole-Parmer Model ~o. J-7003-04 pump eouple~ with motor dri~e Mo~el ~o.
7617-70 Is a ~uitablo pump for the aqueou~ phase.
The output of pump C2 18 connected to feed lIne 150 which 18 then 6pllt at branch f~ttlng 168 Into e~ternal fee~ lIne lC0 and esternal feed l~ne lC4. Llnes 160 and 164 are attache~ to inlet feeds 310C an~ 310D, respect~vely, allow~ng the agueou~
phage to enter mlsIng ehamber 20.
In operation, pump~ 60 and 62 drlve the l~pophIlle phage and agueou~ phase, respectlvely, Into mising chamber 20. ln m~-ing chamber 20, ~hear mlsIng occur~, eau~ing the formatlon of hydrated llpl~ whleh qu~ekly eoalesce~ into multilamellar or paucllamellar lIpi~ ~eslelea. These lIpid ves~el-~are w~thdrawn from mislng ehamber 20 through eslt tube 170. Removal of tho ~es~cle~ may be enhaneed, In eertain Instanee~, by the act~on of pump 6~, preferably a vacuum pump.
Prototyp~cal mlsing chamber~ ha~e been ma~e ln bloek of Luelto brand plagt~e whleh flt- ~nto the palm o the hand. She eyl~n~r~eal Inner ~urfaee of the ehamber ha~ on 8mm ~lameter wlth a 5m~
~iamoter dl~eharge or~fle- 320. In th~ prototype, ~nlet orlf~ee~ 310A-310D eae~ have lmm ro~lu~. In *Trade Mark -17- 1320~00 the teste~ ~ystem, two inlet orifices, 310A-~lOB are fed by the described pump 60 to del$ver the l$pi~
mi~ture to the chamber wh$1e the other two inlet orifices 310C-310D are fe~ by the other describe~
pump 62 which delivers aqueous phase. The absolute an~ relat$ve flow velocities are controlle~ to g$ve lipi~ vesicles of desired properties. Water-soluble materials to bs encapsulate~ are include~ in the agueou~ phase, ana lipoph~lic materials ~n the llpi~
10 phase. ~ip$~ ves$cle~ can be fashionod in le~ than a ~econ~ when the preferre~ l$near ~low rate of t least lOm~ i8 used. Actual flow of the material~
can be ad~usted to optim$ze the proces~. For esample, for an aqueous flow rate of 1.9 l~minute, the l$pid misture flow rate $s selected between a rate of 472ml~mm for low hydration and a rate of 94ml~minute for high hydrat$on.
3. Syrinae Dr$ven System FIGURE 4 ~bows an alternat$ve embodiment for 20 forminq lipi~ vesicles. Instea~ of motor-~riven pumps to dr~ve the fluid, apparatus 200 has a lever arrangement for operating syringes 210 an~ 220.
Rai~ing the lever inject~ the lipid material and the aqueous phase into mising chamber 20 where it meets an~ form~ lipi~ vesicle~.
More specif~cally, syringes 210 and 220, prefersbly of con~ent~onal con~truction, have barrels 222 an~ 224 for hol~ng the lip~ n~ aqueou~ phases, respectively. A~ ~hown, barrel~ 222 n~ 22~ re 30 mounte~ on a ~upport member 22S of a platform 228.
For e~ample, tho barrel~ 222 an~ 224 can be hel~ by clamp~ 229 which are also releasably clampable to the support member 226. Other arrangements incorporating different setting mechanisms, e g., ratcheting, detent and retention elements, will be apparent to one ~ille~ in the art. A horizontal base 230 supports the platform 228, an~, as illustrate~, e~ten~s in ~pace~, generally parallel relation to the support member 226. A ~ertical linking member 232 interconnects the tyringe support member 226 an~ tho base 230, as shown, at a f~rst en~ of each.
Syringes 210 an~ 220 are operate~ by manually rais$ng lever arm 240 As shown, lever arm 240 is pivotally connecte~ at one en~ to lin~ing member 232 by pivot connection 242 which ~erves as a fulcrum and, at the other end, e~ten~s longitudlnally beyond the support member 226, culminating in a handle 244 Illustratea lever arm 240 i~ ~ispo6e~ in the space between the support member 226 an~ the base 230 for vertical angular motion As lever arm 240 is pi~oted upwar~ from its horizontal position, it drives syrinqe plungers 246 an~ 248 into the respecti~e syringe barrels 222 an~ 224, in~ecting the fluid materials contalne~ therein ~nto fee~ tubing 250 an~ 252 for ~elivery to mising chamber 20 A~ illustrtte~, nsar the han~le 242 i~ -biasing spring 260 which provide~ restoring force to ~ri~e lover rm 240 ~ownwardly to it~ prof-rably hori~ontal, rea~y po~it~on, n~ maintain~ l-v-r arm 30 240 1~ th- bori~o~tal position when not actuato~.
-lg- 1320400 ~ever arm 240 can be actuate~ manually, or by an optlonal ~ri~er 256, such as a linear motor.
This ~pparatus provides proportional - ~n~ection of the flui~s from æyringes 210 an~ 220.
As will be apparent to one ækille~ in the art, the estent of displacement of the plunger~ 246 an~ 248 into barrelæ 222 an~ 228 i8 proportional to the vertical ~isplacement of lever arm 240 at plunger~
246 and 24B. Thi~, ln turn, is proportional to the distance from pivot connection 242 to respoctive plunqer~ 246 and 248. Thu~, if all else is egual, by placing the lipi~ material in syringe 210 an~ buffer in syringe 220, more buffer than lipi~ material i8 ~n~ected ~ince syringe 210 iæ closer than syringe 220 to pivot connect~on 242~ By proper placement of the syringes 210 anh 220 along the æupport member 226, a selected ratio of buffer to lipi~ material can be controllably in~ecte~.
Both of the describe~ drive mechanisms can be u~e~ with the Eame mising chamber an~ perform the metho~ of the ~nvention. Other type~ of ~riv-app~ratus an~ mo~ifications in the mising chamber can be use~ in the metho~æ of the in~ention. ~uch other mo~ifications, Including mo~ification~ of the apparatu~, will be read~ly apparent to one ~kille~ in the art. ~uch oth~r mo~ificat~ons are includea w~th~n the following cla~m~.
What ~ cla~mea ~:
vesicles are often classifie~ into three groups by size an~ structure: multilamellar vesicles (~MLV'5~), large unilamellar vesicles ~L W 's~), and small unilamellar vesicles (~S W 's'). ~LV'8 are onion-like structures having a serie~ of substanti~lly spherical shells forme~ of lipid b~layer~ intersperse~ with aqueous layers. L W 's have a ~iameter greater than 1 ~ ana are forme~ of a lipi~ bil~yer ~urroun~ing a 10 large, unstructure~ a~ueous phase. S W 'fi are ~imilar in ~tructure to the L W '~ escept their ~ameters are less than 0.2 p.
A ourth type of lipi~ vesicle, which is partioularly well suite~ for transport of either lipiDs or aqueou~ materials, i~ the paucilamellar vesicle (~PLV~ Q, Callow and McGrath, Cryobiology 1985, ~3), pp. 251-267. This type of vesicle has an e~ternal structure of about two tc five peripheral lipid bilayers with a large, 20 unstructure~ aqueous center. Lipid droplets, e.g., oil droplets, may be suspended in the aqueous center, lea~ing to very high uptake of aqueous or lipophilic materials. Paucilamellar vesicles range from about 2-15 ~ in ~iameter.
E~ch type of l~pi~ vesicle has distin~t advantages for certa~n uses. Because of the relatively large amount of lipi~ ln the lip~
bilayer~ of the NLV's, they are consi~ere~ best for encapsulation or transportation of l~pophille 30 material~. She L W '~, because of the~r large aqueous~lip~ volume rat~o, are consi~ere~ best for .~;, encapsulation of hydrophilic molecules, particularly macromolecules. S W 's have the advantage of small size which allows relatively easy access to the cells of tissue, but their small volume limits delivery o~
hydrophilic aqueous materials to trace amounts.
However, S W 's may be useful in place o MLY'~ for the transportation of small guant~ties of lipophilic material~ because of high lipid~water ratios. PLV's can tran~port largs quantities of agueous or lipophilic material~ but their large size can preclude approach to certain tissues.
The pre~ent invention pertains to the formation of MLV's an~ PLV's. Since SW '~ are commonly made by sonification of multilamellar lipid Yesicles, it follows that the processes by which MLV'~ are produced can be used as part of a technigue for making SW'8.
The conventional approach to producing multilamellar lipid vesicles, particularly liposomes maDe of pho~pholipi~s, ~tarts by difisol~ing the lipid~, together with any lipophilic aaditlves, in an organic solvent. The organic solYent is then removed by evaporation using heat or by passing a stream of an inert gas (e.g., nitrogen) oYer the dissol~e~
lipids. ~he resi~u~ ~s then hydrated with an agueous phase, generally containing electrolytes an~
a~ditives ~uch as hyaroph~lic biologlcally-actiYe materials, to form large multilamellar llpl~ membrane structuro~. In rome ~ariat~ons, ~ifferent types of 30 partlculate mattor or stru~tures have been u~e~
~uring the ovaporation process to assist in the ~ ~3~0400 formation of the lipi~ residue. Those in tbe fiel~
have shown that by changing the physical structure of the lipid resiaue, better vesicles form upon hydration. Two recent review publications, Szoka ana Papahdjopoulo~, Ann. Rev. Biophy6. Bioeng. 2:467-508 (1980), an~ Dousset ana Douste-Blazy ~in LL~sssme~, Puisieus ana Delattre, E~itors, technigue~
et Documentation Lavoi6ier, P-ris, pp.~l-73 ~1985)), summarize the methoa6 which have been use~ to make MLV ' .
~o matter how the MIV'~ or PLV'~ are formea, once maae it i8 necessary to determine th~
effectiveness of the process. ~wo measurement~
commonly usea to determine the effectiveness of encapsulation of materials in lipi~ vesicles are the encapsulatet mass and capturea volume. The encapsulated m~s is the masfi of the substanco encapsulatea per unit mass of the lipia ana ~s often given as a percentage. The capture~ volume i~
defined as the amount of the aqueous phase trappe~
inside the vesicle aiviaea by the amount of lipi~ in the vesi~le structure, normally given in ml ligui~g lipi~.
A ai6a~vantage as~ociatea with proaucing MUV'~ or PLV'~ usinq stan~ard metho~s ~s that these processes are costly, slow an~ relatively ineffic~ent ~n terms of matorial. For eYample, the stan~ard time to manufacture M~Y's ~ ~n the or~er 2-20 hour~. If ~ W '~ ~re rogulre~, the sonication which break~ the multila~ellar lipi~ ~tructures ~nto 8 W '- take~
a~itlonal t~mo. I~iJ ~low processing i~ unwiel~y s 1320~0o - -an~ e~pensive for any large scale production of lipi~
ve~icles .
While rapi~, continuous-flow, mi~ing processes are known in other arts, e.g., chemical kinetics, tbe a~aptation of such processes for the production of lipiD ~esicles has not heretofore been deviseD, or even suggesteD.
Accordingly, an ob~ect of the invention i~
to provi~e an improvea metho~ and apparatus for making multilamellar or paucilamellar lipid vesicle5.
A further object of the invention is to provide a lipid vesicle forming technique which len~s itself to commercial, high-volume production.
Another object of the invention to provide a metho~ and apparatus for the rapid, efficient encapsulat~on o biologically-active macromolecules into lipid vesicles.
These an~ other object~ an~ features of the in~ention will be apparent from the following 20 descriptlon an~ firawings.
~ummar~ of the ~nvention She presen~ invention feature~ a metho~ of pro~ucing multilamellar or paucilamellar lipi~
vesi~le~ and an apparatu8 useful ln thelr proDuction. She metho~ i~ base~ on ths shear mi~ing of a ~olventless lipophilic phase, an~ a hy~rophilic or aqueoug phase ~n order to rapi~ly hy~rate the lipophilic phase, thereby formlng multilamellar or paucilamellar ~tructures. ~he ~ifference in structure is dependent, ln part, on the choice of materials forming tho lipophilic pha~o.
The method of the in~ention uses a mising chamber having a substantially cylin~rical lnner surface with at least ono substantially tangentially locate~ ~nlet orifice which allow flow of one phase, 10 preferably tho lipi~ phase, into the hollow interior. An ad~itional inlet orifice, which may be tangentially or ~ubstantially asially locate~, provi~es input of the other phase, preferably the aqueous phaso. There i8 also a substantially asially locate~ outlet orifice which allows removal of the l~pi~ vesicles after formation.
Th~ l~pophilic phase i~ forme~ of -surfactant, preferably blen~e~ with ~ steroi~ such a5 cholesterol an~ an amphiphilic charge-pro~uc~ng 20 material, a~ well a8 any lipophilic material~ to be incorporated into the lipi~ vesiclo. Thi~ lipophilic phase iB injecte~ into the mising chamber through at least one of the inlet or~fices. An agueous phase, which contain~ primarily water or buffer an~ any hy~roph~l~c material~ to be incorporate~ into tho l~p~ ve~iclo, enter~ the ~ising chsmber through a ~ubstantially tangentially locats~ input orif~ce or an asially locate~ inlet orifico. Both tho l~poph~lic pha~ an~ agueou~ pha~e ar~ in~ecto~ with 30 ~ufficient force to croat- a rap~ ubstantially flo~, ~rsferably tangont~al, about the inner ~urface ~7~ 13 2 0 ~00 of the mi~ing chsmber. The two streams of flowing liquid, the lipophilic phase and the aqueous phase, intersect in such a manner as to cause shear mi~ing, thereby forming the lipid vesicles. Turbulent flow for each liguid i8 preferred but may be unnecessary so long as the two phases intersect with sufficient force. The formed lipid vesicles are removed from the chamber through the asial outlet orifice and collected. In the preferred embodiment of th-lnvention, two tangential flow~ of liquid intersect when travellng in the same direction, e.9., both clockwise or counter-clockwise about the inner surface of the mis~ng vessel. Depending on the pump speed, this flow allows shear mi~ing without causing e~cessive turbulence which would slow down the net flow. ~Shear mising~ is define~ as the mising of the lipophilic phase with the agueous phase un~er turbulent or ~hear con~itions which provide a~eguate mising to hyarate the lipid and form lipid vesicles.
20 The pump speeds are modified depending on the viscosity of the materials and the size of the orifices selected. ~Shear mi~ing~ is achieved by liqui~ shear which i~ substantially eguivalent to a flow rate for the combined phases of 5-30 m~s through a 1 mm radius oriflce. In Dnother embodiment of the invention, the two ~treams of fluid intersect by traveling in countercurrent directions, e.g., one clockwlse and one counter-clockw~e. Thl~ 18 most u~eful fo~ very dilute ~olutions where escea~i~e 30 turbulence and resultin~ back pressure is not as much of a problem.
-8- 1~ 20~100 Preferred surfactants for use ~n the ~nvention lnclude polyosyethylene acyl ether~, polyo~yethylene acyl amine~, polyosyethylene alkyl esters, polyosyethylene ~orbita~ alkyl esters, and polyosyethylene glycerol alkyl ester~, where sa~d ~
alkyl or acyl groups have 16-20 carbon atoms and not more than two ~ouble bon~g. Many of these materials are liquid at room temperature ~o no heatlng or other treatment ~tep is necessary to achieve the fluld 10 stage helpful for rap~ flow. If the mater1als are not liguid at operating temperature, heating befor~
introduction through the tangential inlet orifice ls a preferre~ step since the liqul~ state provi~es proper flow about the mising chamber.
Selection of the proper surfactant can modify the structura of the final lipid vesicles.
Certain surfactants will form PLV's wh~le others will form MLV's. Although not necessary to the invention, the difference 1~ theorized to occur because of the 20 ~ifferences in the lipophilic~hydrophilic balance of the surfactants.
The chamber should be ma~e of materials whlch can withstand ~olut~ons whlch are heated above the melting point of the lipi~. A chamber which will withstand ~0C. ~a temperature high enough to melt ~ubstantially all of the llpids within thls class) ~s helpful.
Th~ inv-ntlon al80 feature~ a ~evlce or ma~lng llp1d v-~icles. Th~ central part of thls 30 de~ice 1s the mlsing chamber w~th the inputs as -9_ 1320400 previously descri~ed. In addition, this device should have reservoirs for the lipophilic and hydrophilic phases an~ pumps or drive mechanisms which force the phases, preferably in fluid form, - through the inlet orifices into the mising chamber.
In addition, the inlet path from the reservoirs, or optionally the reservoirs themselves, can include mising device insuring uniform dispersions of each of the lipophilic an~ ~gueous phases. Another option i8 to have heaters, either withln the reserYoir or between the reser~oir and the mising chamber, to allow heating of the phases if necessary for proper liguid flow.
The ~pecifics of the pumps, feea lines, and orifice ~ize selection are a matter of engineering within the skill of those practicing the art and depend on amount an~ viscosity of the material to be processed and the desired flow rates.
These an~ other features of the invention 20 will be more apparent from the drawings and the det~iled description.
Brief Description of the Drawins~
FI¢URE 1 i~ an perspect~ve view of 8 mising chamber for use ~n accordance with the pr~ctice of the ~nvent~on;
FIGURE 2 ~ a ~ectional v~ew taken along l~ne I-I of the mising chamber of FIGURE l;
~o 1320~00 FIGURE 3 i~ a schematic representation of a lipi~ vesicle producing apparatus in accordance with a first embodiment of the invention; an~
FIGURE ~ i8 a schematic representation of a lipi~ vesicle pro~ucing apparatus in accordance with a ~econ~ embo~iment of the invention.
Detaile~ Description of Preferre~ Embo~iment~
As note~ previously, the present invention features a metho~ of making multilsmellar or lO paucilamellar l~pi~ ~e~icle~ an~ a ~evice for the manufacture of those lipid vesicles. The metho~s an~
~evice have a mising chamber with a substantially cylin~rical interior Qurface as a central feature.
This mi~ing chsmber permits input of material~ with controlled intersection of the lipophilic an~
hy~rophil~c phase~ ~n or~er to form the multilamellar or paucilamellar lipi~ ve~icles. ~he tsrm ~substantially cylin~rical~ as use~ herein means an~
~nclu~es cylin~r~cal, frustoconical an~ other ~hapes 20 having substantially similar surfaces of re~olution.
The following description will more clearly eluci~ate the principleR of the invention.
l. The M~ina Chamber A mising chamber ~uitable $n the pre~ent ~n~entlon ~ ~hown ln FIGUXE l. Mi~ng chamber 20 h~ a ~ub~tant~-lly cyl~n~rical cro~-section form~ng ~ central, hollowe~ out portion of a block of -ll- 1320~00 material 300. Inlet feed lines, illustrateD as 310A-310D, allow the lipophilic anD agueous phases to be in~ecte~ into mising chamber 20. Each of these inlet feed lines 310A-310D ends in an inlet orifice 320A-320D.
FIGURE 2 18 a cross-sectional illustration of mising chamber 20 about lin~ I-I. As shown in FIGURES 1 anD 2, mi~ing chamber 20 has a cylin~rical inner ~urfac~ 330 DisposeD about a central asis ~Desi9nateD ~c~) anD i8 close~ at each enD by a circular face 332 anD 334. Discharge orifice 350 ~s disposeD through one of the circular faces, for esample, as shown, through face 332. Discharge orifice 350, which preferably has of circular cross-section centereD on the central asis c, ha~ a ~iameter ~ubstantially less than that of the cylinDrical surface 330. In this preferre~
embodiment, discharge orifice 350 i~ both asially and raDially spaced from the inlet orifices 320A-320D.
Inlet orifices 320A-320D ser~e as in~ection ~ets anD are arrangea in paceD relation perpherally about mising chamber 20. While it is preferreD that inlet orifices 320A-320D, as well as inlet fee~ lines 310A-310D, are arrangeD in a plane ~ispo~eD
perpenD~cular to the central asi~ c of the cylinDrical mlsing chamber 20, th~ ~s not requireD.
The four ~nlet orifices 320A-320D shown ~n FIGURE 1 are ~paeeD about the cy~in~rical inner ~urface 330 at nin~ty ~egres ~ntor~al~ anD are tangential relati~e 30 to cylinDrical innor ~urace 330.
-12- 13~0~00 Lipi~ material is introduced into mising chamber 20 through inlet orifices 320A and 320B which are connectea to esternal feed lines 158 and 162 (F~GURE 3) through inlet feed lines 310A-3108 ~t diametrically oppo~ite si~es of cylindrical ~urfsce 330. The agueous phase i8 introducea through inlet orifices 320C an~ 320D which are connecte~ to esternal feed lines lC0 and 164 ~FIGU~E 3) through inlet feed lines 310C an~ 310D at di~metrically opposite ~ides of cylindrical ~urface 330. With the illustrated arrangement, the fluids flow rotationally, as indicated by ~rrow ~a~, within the mising chamber 20. All the fluids flow in the ~ame direction, either clockwi~e or counter-clockwise, depending on whether viewed from face 332 or face 334.
The flow rate~, volumes and particularly the flow paths are key~to the efficient production of quality vesicles according to the invention. The fluids enter the mi~ing chamber 30 tanqentially with respect to cylindrical wall 330, ~pprosimately 90-from the other inlet orifices. The flui~ stream 1 guide~ ~n ~ ~piral flow path from each ~n~ect~on orifiee 310A-310D arouna the inner ~urface of the cylindrical w~ll 330, ~nd then out through d~scharge oriflco 350 ~nto an oslt tube 170. $he flow p~th~
are controlled by the or~ent~tion ~nd pl~cement of the ~n~ection orifices 310A-310D 80 a8 to intersect, creating the mi-~ng rone. The pump ~pee~, a~ well as the orifice an~ fee~ l~ne ~iameters, are ~electea 30 to achieve proper ~hear mising for l~pi~ vesicle format~on. In mo~t c~rcumstance~, turbulent flow rolecte~ to prov~e ~equ~to mis~ng.
2. Pump Dr~ven Flui~ DeviGg As note~ previously, the mising chamber is the heart of the apparatus for making the multilamellar or paucilamellar lipi~ vesicle~.
FIGURE 3 illustrates one embodiment of the invent~on which uses th~ mising chamber.
~ i~ing chamber 20 (not shown) i~ within block 300. She lipid reservoir 40 hol~s the lipophilic pha~e an~ the agueous reservoir S0 hol~s 10 the agueous phase. Connected to the reservoir~ an~
the outlet from the mising chamber are the first, second and third pump~ 60, 62 and 64 with their respective controller~ 70, 72 an~ 74. Thir~ pump 64 and it~ controller 74 are optional and are unnece~sary un~er most circumstance~.
The lipophilic phase, consisting the surfactant an~ the other lipophilic materials ~e.g., a steroid such a~ chole~terol, a charge-pro~ucinq amphiphile, an~ any lipophilic materials to be 20 encapsulated) are blended and placed in lipi~
reservoir 40. Heater 30 ~ optional an~ i8 meant to illustrate any device which can preheat the lipophilic phass (if neede~) to allow it to flow a~ a flui~ at tempersture of operation. Heater 30 may be a hot plate, an oven, or the entlre apparatus may be place~ in a warm room.
Preferably, the temperature of the lip~
material within hol~ing reservoir 40 i8 such that the lipi~ material remain~ ~n a flui~ ~tate. For lipi~
-l~- 1320400 materials with melting points above room temperature, ~t maybe necessary to keep the material at elevate~ -temperature within the reservoir. So that en~, holding reservoir 40 can serve as an incubator, optionally having a warming element such as tho illustrate~ heater coll 106 which is helically wound about the interior holding ves~el 102, and~or -thermally insulating ~ac~et enclosing hol~ing vess-l 102. ~n certain instance~, it 1~ ~esirable to melt the lipid material directly within holding reservolr 40 by employing warming element 106, thereby eliminating the nee~ for separate heating ~evice 30.
If the lipi~ is a ligui~ at room temperature, no heating device i~ necessary.
- Lipophilic additives, e.g., steroi~s such as cholesterol, are normally blended into the lipid before ths lipophilic phase reaches holding reservoir 40. However, additives can be introduced ~irectly into holding reservoir 40 and blendea into ths 20 lipid. If 80, a mising device such 8S a magnetic or mechanical miser 108 is provide~. Illustrated miser 108 has a vaned, mising element 110 e~ten~ing longitu~inally within the vessel 102. Mising element 110 is rotationally driven by a driver, such a~ a motor or an electromagnetic driving coil 112 disposed outside holding ves~el 102, with which mising element 110 i~ magnetically couple~.
Lipi~ re~ervoir 40 is coupled to the inlet of pump 60 by tubing 132. Preferably, pump 60 i8 a 30 po~itive ~l~placement, magnetically ~riven pump. lt ~ al~o preferred that the pump be detachable from -15- 1~20400 ~t~ dr~ving motor~ (not ~eparately ~hown) for steril~ing, for esample, by autoclav~ng. A ~u~table pump for the l~p~ mater~al, for e~ample, ~ made by Cole-Parmer, Model ~o. J-7002-23, coupled with motor drive Model No. 7144-0~.
The output from pump C0 ~ connected to ml~lng chamber 20 by fee~ line 150. Feed l~ne 150 ~8 provided wlth a flow ~plitting or branch f~ttlng arran~ement lC6 whereby fir~t llne lS~ 1~ connect~ad to both estern~l feed llne 158 and esternal feed llne 162. Feed line 150 ha~, for esample, a 1~4 lnch (0.1 cm) outer dlameter and a 1~8 lnch ~0.05 cm) inner dlameter. All connectlons are pre~era~ly made with compression fitting~, for esample, of Teflon brand material which can be autoclaved at the ~ame time ~s the lines.
Each of the esternal feed llnes 158 ~n~ 162 are connecte~ to inlet fee~ llnes for the l~poph~lic phase, specific~lly inlet feed lines 310A and 310B, 20 respectively.
FIGURE 3 al~o illustrates the agueous phase portion of the system. Aqueous reservoir S0 ~
~imilar ln construction to llpi~ re~ervolr ~0 escept cham~er 122 ~ 8 larger ~ecauso the aqueou~ volume ~dde~ to hydrate the lipophilic pha~e i~ normally much larger than the llpi~ volu~e. Neater 126 ~
optional an~ ia used normally if the agueou~ phase ~s heate~. Thi~ permlt~ the reaction to ta~e pl~ce witbout largc temperature aifference~ which provi~e~
30 better lipl~ vesicl~ formatlon.
*Trade Mark -16- 1320~00 The aqueoua ph~se, ~nclud~ng any a~t~vea or b~ologicals which are to be encapsulated, ~
place~ In agueou~ reservo~r 50 an~ flow~ through tubing 13~ to pump 62. Aga~n, pump 62 Ig preferably a positlve d~splacement, magnetieally drl~en pump w~th a detachable ~rl~e motor. A Cole-Parmer Model ~o. J-7003-04 pump eouple~ with motor dri~e Mo~el ~o.
7617-70 Is a ~uitablo pump for the aqueou~ phase.
The output of pump C2 18 connected to feed lIne 150 which 18 then 6pllt at branch f~ttlng 168 Into e~ternal fee~ lIne lC0 and esternal feed l~ne lC4. Llnes 160 and 164 are attache~ to inlet feeds 310C an~ 310D, respect~vely, allow~ng the agueou~
phage to enter mlsIng ehamber 20.
In operation, pump~ 60 and 62 drlve the l~pophIlle phage and agueou~ phase, respectlvely, Into mising chamber 20. ln m~-ing chamber 20, ~hear mlsIng occur~, eau~ing the formatlon of hydrated llpl~ whleh qu~ekly eoalesce~ into multilamellar or paucllamellar lIpi~ ~eslelea. These lIpid ves~el-~are w~thdrawn from mislng ehamber 20 through eslt tube 170. Removal of tho ~es~cle~ may be enhaneed, In eertain Instanee~, by the act~on of pump 6~, preferably a vacuum pump.
Prototyp~cal mlsing chamber~ ha~e been ma~e ln bloek of Luelto brand plagt~e whleh flt- ~nto the palm o the hand. She eyl~n~r~eal Inner ~urfaee of the ehamber ha~ on 8mm ~lameter wlth a 5m~
~iamoter dl~eharge or~fle- 320. In th~ prototype, ~nlet orlf~ee~ 310A-310D eae~ have lmm ro~lu~. In *Trade Mark -17- 1320~00 the teste~ ~ystem, two inlet orifices, 310A-~lOB are fed by the described pump 60 to del$ver the l$pi~
mi~ture to the chamber wh$1e the other two inlet orifices 310C-310D are fe~ by the other describe~
pump 62 which delivers aqueous phase. The absolute an~ relat$ve flow velocities are controlle~ to g$ve lipi~ vesicles of desired properties. Water-soluble materials to bs encapsulate~ are include~ in the agueou~ phase, ana lipoph~lic materials ~n the llpi~
10 phase. ~ip$~ ves$cle~ can be fashionod in le~ than a ~econ~ when the preferre~ l$near ~low rate of t least lOm~ i8 used. Actual flow of the material~
can be ad~usted to optim$ze the proces~. For esample, for an aqueous flow rate of 1.9 l~minute, the l$pid misture flow rate $s selected between a rate of 472ml~mm for low hydration and a rate of 94ml~minute for high hydrat$on.
3. Syrinae Dr$ven System FIGURE 4 ~bows an alternat$ve embodiment for 20 forminq lipi~ vesicles. Instea~ of motor-~riven pumps to dr~ve the fluid, apparatus 200 has a lever arrangement for operating syringes 210 an~ 220.
Rai~ing the lever inject~ the lipid material and the aqueous phase into mising chamber 20 where it meets an~ form~ lipi~ vesicle~.
More specif~cally, syringes 210 and 220, prefersbly of con~ent~onal con~truction, have barrels 222 an~ 224 for hol~ng the lip~ n~ aqueou~ phases, respectively. A~ ~hown, barrel~ 222 n~ 22~ re 30 mounte~ on a ~upport member 22S of a platform 228.
For e~ample, tho barrel~ 222 an~ 224 can be hel~ by clamp~ 229 which are also releasably clampable to the support member 226. Other arrangements incorporating different setting mechanisms, e g., ratcheting, detent and retention elements, will be apparent to one ~ille~ in the art. A horizontal base 230 supports the platform 228, an~, as illustrate~, e~ten~s in ~pace~, generally parallel relation to the support member 226. A ~ertical linking member 232 interconnects the tyringe support member 226 an~ tho base 230, as shown, at a f~rst en~ of each.
Syringes 210 an~ 220 are operate~ by manually rais$ng lever arm 240 As shown, lever arm 240 is pivotally connecte~ at one en~ to lin~ing member 232 by pivot connection 242 which ~erves as a fulcrum and, at the other end, e~ten~s longitudlnally beyond the support member 226, culminating in a handle 244 Illustratea lever arm 240 i~ ~ispo6e~ in the space between the support member 226 an~ the base 230 for vertical angular motion As lever arm 240 is pi~oted upwar~ from its horizontal position, it drives syrinqe plungers 246 an~ 248 into the respecti~e syringe barrels 222 an~ 224, in~ecting the fluid materials contalne~ therein ~nto fee~ tubing 250 an~ 252 for ~elivery to mising chamber 20 A~ illustrtte~, nsar the han~le 242 i~ -biasing spring 260 which provide~ restoring force to ~ri~e lover rm 240 ~ownwardly to it~ prof-rably hori~ontal, rea~y po~it~on, n~ maintain~ l-v-r arm 30 240 1~ th- bori~o~tal position when not actuato~.
-lg- 1320400 ~ever arm 240 can be actuate~ manually, or by an optlonal ~ri~er 256, such as a linear motor.
This ~pparatus provides proportional - ~n~ection of the flui~s from æyringes 210 an~ 220.
As will be apparent to one ækille~ in the art, the estent of displacement of the plunger~ 246 an~ 248 into barrelæ 222 an~ 228 i8 proportional to the vertical ~isplacement of lever arm 240 at plunger~
246 and 24B. Thi~, ln turn, is proportional to the distance from pivot connection 242 to respoctive plunqer~ 246 and 248. Thu~, if all else is egual, by placing the lipi~ material in syringe 210 an~ buffer in syringe 220, more buffer than lipi~ material i8 ~n~ected ~ince syringe 210 iæ closer than syringe 220 to pivot connect~on 242~ By proper placement of the syringes 210 anh 220 along the æupport member 226, a selected ratio of buffer to lipi~ material can be controllably in~ecte~.
Both of the describe~ drive mechanisms can be u~e~ with the Eame mising chamber an~ perform the metho~ of the ~nvention. Other type~ of ~riv-app~ratus an~ mo~ifications in the mising chamber can be use~ in the metho~æ of the in~ention. ~uch other mo~ifications, Including mo~ification~ of the apparatu~, will be read~ly apparent to one ~kille~ in the art. ~uch oth~r mo~ificat~ons are includea w~th~n the following cla~m~.
What ~ cla~mea ~:
Claims (20)
1. A method of producing multilamellar or paucilamellar lipid vesicles comprising the steps of:
providing a mixing chamber, said mixing chamber having a substantially cylindrical inner surface having with at least one tangentially directed inlet orifice and at least one additional inlet orifice opening through said inner surface into said mixing chamber, said mixing chamber further including an outlet orifice for removing said lipid vesicles after formation, inputting a lipophilic phase into said mixing chamber through one of said inlet orifices with sufficient force to create a flow of said lipophilic phase about said inner surface, inputting an aqueous phase through another of said inlet orifices with sufficient force to create a flow of said aqueous phase about said inner surface, wherein at least one of said aqueous phase and said lipophilic phase is input through said tangentially directed inlet orifice, allowing said flow of said lipophilic phase and said flow of said aqueous phase to intersect, thereby providing shear mixing of said lipophilic phase and said aqueous phase, said shear mixing providing sufficient mixing to form said lipid vesicles, and removing said lipid vesicles from said mixing chamber through said outlet orifice.
providing a mixing chamber, said mixing chamber having a substantially cylindrical inner surface having with at least one tangentially directed inlet orifice and at least one additional inlet orifice opening through said inner surface into said mixing chamber, said mixing chamber further including an outlet orifice for removing said lipid vesicles after formation, inputting a lipophilic phase into said mixing chamber through one of said inlet orifices with sufficient force to create a flow of said lipophilic phase about said inner surface, inputting an aqueous phase through another of said inlet orifices with sufficient force to create a flow of said aqueous phase about said inner surface, wherein at least one of said aqueous phase and said lipophilic phase is input through said tangentially directed inlet orifice, allowing said flow of said lipophilic phase and said flow of said aqueous phase to intersect, thereby providing shear mixing of said lipophilic phase and said aqueous phase, said shear mixing providing sufficient mixing to form said lipid vesicles, and removing said lipid vesicles from said mixing chamber through said outlet orifice.
2. The method of claim 1 wherein said liopohilic phase comprises a surfactant.
3. The method of claim 2 wherein said surfactant is selected from a group consisting of -polyoxyethylene acyl ethers, polyoxyethlene acyl amines, polyoxyethylene alkyl esters, polyoxyethylene sorbitan alkyl esters, and polyoxyethylene glycerol alkyl esters, where said alkyl or acyl groups have 16-20 carbon atoms atoms not more than two double bonds.
4. The method of claim 2 wherein said lipophilic phase further comprises a steroid selected from a group consisting of cholesterol, hydrocortisone and their analogs or chemical equivalents.
5. The method of claim 1 wherein said inlet orifice for said aqueous phase is located axially and removed from said outlet orifice.
6. The method of claim 1 wherein said inlet orifices for said aqueous and lipophilic phases are both located tangentially and said phases have tangential flow about said inner surface.
7. The method of claim 6 wherein said tangential flow of said lipophilic phase and said tangential flow of said aqueous phase are co-directional.
8. The method of claim 2 further comprising the step of liquifying said lipophilic phase by heating said lipophilic phase to a temperature above the melting point of said surfactant before introduction of said lipiohilic phase into said mixing chamber.
9. The method of claim 8 further comprising the step of heating said aqueous phase to a temperature above the melting point of said surfactant before introduction of said aqueous phase into said mixing chamber.
10. The method of claim 1 wherein a lipophilic material is encapsulated in said lipid vesicles by incorporation of said lipophilic material into said lipophilic phase before introduction of said lipophilic phase into said mixing chamber.
11. An apparatus for the production of multilamellar or paucilamellar lipid vesicles by shear mixing of a lipophilic phase and a hydrophilic phase, said apparatus comprising:
a substantially hollow mixing chamber having an interior surface, an exterior surface, and a cross-section which is substantially cylindrical about a first axis, said mixing chamber further containing a first inlet orifice for inlet of said lipophilic phase, a second inlet orifice, for inlet of said hydrophilic phase, and an outlet orifice, said first inlet orifice bing located substantially tangential to said inner surface of said mixing chamber;
a first reservoir for holding said lipophilic phase:
a first delivery system for delivering said lipophilic phase from said first reservoir to said first inlet orifice;
a second reservoir for holding said hydrophilic phase; and a second reservoir system for delivering said hydrophilic phase from said second reservoir to said second said inlet orifice.
a substantially hollow mixing chamber having an interior surface, an exterior surface, and a cross-section which is substantially cylindrical about a first axis, said mixing chamber further containing a first inlet orifice for inlet of said lipophilic phase, a second inlet orifice, for inlet of said hydrophilic phase, and an outlet orifice, said first inlet orifice bing located substantially tangential to said inner surface of said mixing chamber;
a first reservoir for holding said lipophilic phase:
a first delivery system for delivering said lipophilic phase from said first reservoir to said first inlet orifice;
a second reservoir for holding said hydrophilic phase; and a second reservoir system for delivering said hydrophilic phase from said second reservoir to said second said inlet orifice.
12. The apparatus of claim 11 wherein said outlet orifice is located substantially axially about said first axis.
13. She apparatus of claim 12 wherein said second inlet orifice is located substantially axially about said first axis at a position removed from said outlet orifice.
14. The apparatus of claim 12 wherein said second inlet orifice is located substantially tangential to said inner surface of said mixing chamber.
15. The apparatus of claim 11 wherein said first delivery system comprises a first pump, a first inlet feed line connecting said first pump, a first pump to said first inlet orifice, and a first tubing connecting first reservoir to said first pump.
16. The apparatus of claim 15 wherein said second delivery system comprises a second pump, a second inlet feed line connecting said second pump to said second inlet orifice, and a second tubing connecting said second reservoir to said second pump.
17. She apparatus of claim 11 wherein said first reservoir comprises a first holding tank and a first heater.
18. The apparatus of claim 11 wherein said second reservoir comprises a second holding tank and a second heater.
19. The apparatus of claim 14 wherein said mixing chamber comprises a plurality of said first inlet orifices and a plurality of said second inlet orifices.
20. The apparatus of claim 16 wherein said first and second pumps, said first and second tubing, said first and second inlet feed lines, and said first and second inlet orifices are selected in size to provide shear mixing
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US163,806 | 1988-03-03 | ||
| US07/163,806 US4895452A (en) | 1988-03-03 | 1988-03-03 | Method and apparatus for producing lipid vesicles |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA1320400C true CA1320400C (en) | 1993-07-20 |
Family
ID=22591653
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA000592763A Expired - Lifetime CA1320400C (en) | 1988-03-03 | 1989-03-03 | Method and apparatus for producing lipid vesicles |
Country Status (9)
| Country | Link |
|---|---|
| US (1) | US4895452A (en) |
| EP (1) | EP0406273B1 (en) |
| JP (1) | JP2542271B2 (en) |
| KR (1) | KR900700072A (en) |
| AU (1) | AU614598B2 (en) |
| BR (1) | BR8907293A (en) |
| CA (1) | CA1320400C (en) |
| DE (1) | DE68910969T2 (en) |
| WO (1) | WO1989007929A1 (en) |
Families Citing this family (108)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1992011083A1 (en) * | 1987-09-28 | 1992-07-09 | Redding Bruce K Jr | Apparatus and method for making microcapsules |
| US5271881A (en) * | 1987-09-28 | 1993-12-21 | Redding Bruce K | Apparatus and method for making microcapsules |
| US5013497A (en) * | 1988-03-03 | 1991-05-07 | Micro-Pak, Inc. | Method and apparatus for producing lipid vesicles |
| FR2649335B1 (en) * | 1989-07-05 | 1991-09-20 | Texinfine Sa | METHOD AND DEVICE FOR THE DIRECT PRODUCTION OF LIPOSOMES |
| US6407061B1 (en) * | 1989-12-05 | 2002-06-18 | Chiron Corporation | Method for administering insulin-like growth factor to the brain |
| US5624898A (en) * | 1989-12-05 | 1997-04-29 | Ramsey Foundation | Method for administering neurologic agents to the brain |
| US5230253A (en) * | 1990-02-22 | 1993-07-27 | Beckman Instruments, Inc. | Fluid mixing device |
| DE4031094A1 (en) * | 1990-10-02 | 1992-04-09 | Basf Ag | METHOD FOR PRODUCING STABLE INJECTABLE (BETA) CAROTINE SOLUBILISATES |
| US5256422A (en) * | 1991-03-28 | 1993-10-26 | Micro Vesicular Systems, Inc. | Lipid vesicle containing water-in-oil emulsions |
| US5326165A (en) * | 1991-06-26 | 1994-07-05 | Irvine Scientific Sales Co. | Mixing apparatus |
| US5523063A (en) * | 1992-12-02 | 1996-06-04 | Applied Materials, Inc. | Apparatus for the turbulent mixing of gases |
| CA2150463A1 (en) * | 1992-12-02 | 1994-06-09 | Donald F. H. Wallach | Extended duration antacid product |
| JPH08512056A (en) * | 1993-06-30 | 1996-12-17 | ジェネンテク・インコーポレイテッド | Method for producing liposomes |
| WO1995016437A1 (en) * | 1993-12-17 | 1995-06-22 | Micro-Pak, Inc. | Method of transmitting a biologically active material to a cell |
| DE19507410C2 (en) * | 1995-03-03 | 1997-05-22 | Gsf Forschungszentrum Umwelt | Method and device for producing aerosols |
| US5662957A (en) * | 1996-05-03 | 1997-09-02 | Novavax, Inc. | Oil containing lipid vesicles with marine applications |
| US5700679A (en) * | 1996-06-07 | 1997-12-23 | Novavax, Inc. | Lipid vesicles having a bilayer containing a surfactant with anti-viral and spermicidal activity |
| US5846551A (en) * | 1996-06-10 | 1998-12-08 | E-L Management Corp. | Water-based makeup compositions and methods for their preparation |
| US5776536A (en) * | 1996-12-23 | 1998-07-07 | Igen, Inc. | Reduced fat chocolate and method of manufacture |
| US6039960A (en) * | 1997-05-28 | 2000-03-21 | E-L Management Corp. | Water containing wax-based product |
| TW476788B (en) | 1998-04-08 | 2002-02-21 | Kimberly Clark Co | A cleanser and the making process thereof |
| US6193997B1 (en) * | 1998-09-27 | 2001-02-27 | Generex Pharmaceuticals Inc. | Proteinic drug delivery system using membrane mimetics |
| US6855296B1 (en) | 1998-11-13 | 2005-02-15 | Optime Therapeutics, Inc. | Method and apparatus for liposome production |
| IL143104A (en) * | 1998-11-13 | 2005-09-25 | Optime Therapeutics Inc | Method and apparatus for liposome production |
| US7273618B2 (en) * | 1998-12-09 | 2007-09-25 | Chiron Corporation | Method for administering agents to the central nervous system |
| US6080211A (en) * | 1999-02-19 | 2000-06-27 | Igen, Inc. | Lipid vesicle-based fuel additives and liquid energy sources containing same |
| US6559189B2 (en) | 1999-04-28 | 2003-05-06 | Regents Of The University Of Michigan | Non-toxic antimicrobial compositions and methods of use |
| US6635676B2 (en) | 1999-04-28 | 2003-10-21 | Regents Of The University Of Michigan | Non-toxic antimicrobial compositions and methods of use |
| US7655252B2 (en) | 1999-04-28 | 2010-02-02 | The Regents Of The University Of Michigan | Antimicrobial nanoemulsion compositions and methods |
| US7767216B2 (en) * | 1999-04-28 | 2010-08-03 | The Regents Of The University Of Michigan | Antimicrobial compositions and methods of use |
| US6506803B1 (en) | 1999-04-28 | 2003-01-14 | Regents Of The University Of Michigan | Methods of preventing and treating microbial infections |
| US8236335B2 (en) * | 1999-04-28 | 2012-08-07 | The Regents Of The University Of Michigan | Antimicrobial nanoemulsion compositions and methods |
| JP2003504390A (en) * | 1999-07-15 | 2003-02-04 | イネックス ファーマスーティカルズ コーポレイション | Method and apparatus for the production of lipid vesicles |
| US6309664B1 (en) | 1999-09-15 | 2001-10-30 | Igen, Incorporated | Methods, uses and compositions of fluid petrolatum |
| EP1203614A1 (en) * | 2000-11-03 | 2002-05-08 | Polymun Scientific Immunbiologische Forschung GmbH | Process and apparatus for preparing lipid vesicles |
| EP1528937B1 (en) * | 2001-06-05 | 2016-08-10 | The Regents Of The University Of Michigan | Nanoemulsion vaccines |
| AU2003230600B2 (en) * | 2002-03-05 | 2009-06-04 | Transave, Inc. | An inhalation system for prevention and treatment of intracellular infections |
| US6899890B2 (en) * | 2002-03-20 | 2005-05-31 | Kv Pharmaceutical Company | Bioadhesive drug delivery system |
| ES2354607T3 (en) | 2002-06-28 | 2011-03-16 | Protiva Biotherapeutics Inc. | PROCEDURE AND APPLIANCE TO PRODUCE LIPOSOMES. |
| KR101301653B1 (en) * | 2002-10-29 | 2013-08-30 | 트랜세이브, 인코포레이티드 | Sustained release of antiinfectives |
| US7718189B2 (en) * | 2002-10-29 | 2010-05-18 | Transave, Inc. | Sustained release of antiinfectives |
| US7879351B2 (en) | 2002-10-29 | 2011-02-01 | Transave, Inc. | High delivery rates for lipid based drug formulations, and methods of treatment thereof |
| US20040087564A1 (en) * | 2002-10-31 | 2004-05-06 | Wright D. Craig | Delivery composition and method |
| US20050208083A1 (en) | 2003-06-04 | 2005-09-22 | Nanobio Corporation | Compositions for inactivating pathogenic microorganisms, methods of making the compositons, and methods of use thereof |
| US20050100601A1 (en) * | 2003-11-07 | 2005-05-12 | Viratox, L.L.C. | Virucidal activities of cetylpyridinium chloride |
| PT103072B (en) * | 2004-02-13 | 2009-12-02 | Faculdade De Engenharia Da Uni | MIXER IN NETWORK AND RESPECTIVE MIXING PROCESS |
| WO2006071659A1 (en) * | 2004-12-29 | 2006-07-06 | Trustees Of Boston University | Delivery of h2 antagonists |
| US20070054834A1 (en) * | 2005-04-11 | 2007-03-08 | Nanobio Corporation | Quaternary ammonium halides for treatment of infectious conditions |
| JP5639338B2 (en) * | 2005-07-27 | 2014-12-10 | プロチバ バイオセラピューティクス インコーポレイティッド | Liposome production system and production method |
| US20070036831A1 (en) * | 2005-08-09 | 2007-02-15 | Nanobio Corporation | Nanoemulsion compositions having anti-inflammatory activity |
| US7797337B2 (en) * | 2005-09-29 | 2010-09-14 | Scenera Technologies, Llc | Methods, systems, and computer program products for automatically associating data with a resource as metadata based on a characteristic of the resource |
| JP2009510168A (en) | 2005-10-03 | 2009-03-12 | マーク エー． ピンスカイ | Compositions and methods for improved skin care |
| EP3067047B1 (en) | 2005-12-08 | 2022-04-20 | Insmed Incorporated | Lipid-based compositions of antiinfectives for treating pulmonary infections |
| US10138279B2 (en) | 2006-04-13 | 2018-11-27 | Regents Of The University Of Michigan | Compositions and methods for Bacillus anthracis vaccination |
| JP4771151B2 (en) * | 2006-04-28 | 2011-09-14 | 公立大学法人大阪府立大学 | Micro mixer |
| EP2023727B1 (en) | 2006-05-17 | 2016-04-13 | Albert Einstein College of Medicine, Inc. | Prostaglandin transporter inhibitors |
| CA2652362C (en) * | 2006-05-19 | 2014-10-07 | Viroblock S.A. | A composition for inactivating an enveloped virus |
| JP2010516695A (en) * | 2007-01-18 | 2010-05-20 | マーク エー． ピンスカイ | Materials and methods for delivering antioxidants into the skin |
| WO2008103416A1 (en) * | 2007-02-21 | 2008-08-28 | Capps Charles L | Synergistic enhancement of calcium propionate |
| EP2152304B1 (en) | 2007-05-02 | 2018-08-22 | The Regents of the University of Michigan | Nanoemulsion therapeutic compositions and methods of using the same |
| WO2008137717A1 (en) | 2007-05-04 | 2008-11-13 | Transave, Inc. | Compositions of multicationic drugs for reducing interactions with polyanionic biomolecules and methods and uses thereof |
| US9333214B2 (en) | 2007-05-07 | 2016-05-10 | Insmed Incorporated | Method for treating pulmonary disorders with liposomal amikacin formulations |
| US9114081B2 (en) | 2007-05-07 | 2015-08-25 | Insmed Incorporated | Methods of treating pulmonary disorders with liposomal amikacin formulations |
| US9119783B2 (en) | 2007-05-07 | 2015-09-01 | Insmed Incorporated | Method of treating pulmonary disorders with liposomal amikacin formulations |
| US10251839B2 (en) * | 2008-01-22 | 2019-04-09 | Igi Laboratories, Inc. | Lipid vesicles derived from olive oil fatty acids |
| CN102083415A (en) * | 2008-04-18 | 2011-06-01 | 纳米生物公司 | Methods for treating herpes virus infections |
| CA2721800A1 (en) * | 2008-04-21 | 2009-10-29 | Nanobio Corporation | Nanoemulsion influenza vaccine |
| AU2009240389A1 (en) * | 2008-04-25 | 2009-10-29 | Nanobio Corporation | Nanoemulsions for treating fungal, yeast and mold infections |
| WO2009132343A1 (en) * | 2008-04-25 | 2009-10-29 | Nanobio Corporation | Nanoemulsions for treating onchomycosis |
| US9415006B2 (en) * | 2008-05-23 | 2016-08-16 | The Regents Of The University Of Michigan | Immunogenic compositions comprising nanoemulsion and hepatitis B virus immunogen and methods of using the same |
| US20090291095A1 (en) * | 2008-05-23 | 2009-11-26 | The Regents Of The University Of Michigan | Nanoemulsion adjuvants |
| EP2349209A2 (en) | 2008-09-26 | 2011-08-03 | Nanobio Corporation | Nanoemulsion therapeutic compositions and methods of using the same |
| CA2743904A1 (en) | 2008-11-17 | 2010-05-20 | The Regents Of The University Of Michigan | Cancer vaccine compositions and methods of using the same |
| JP2012509932A (en) * | 2008-11-26 | 2012-04-26 | ビロブロック エスアー | Method for inhibiting gram-positive bacteria using nonphospholipid lipid vesicles |
| EP2391342A2 (en) * | 2009-01-28 | 2011-12-07 | Nanobio Corporation | Compositions for treatment and prevention of acne, methods of making the compositions, and methods of use thereof |
| US8668911B2 (en) | 2009-05-14 | 2014-03-11 | The Regents Of The University Of Michigan | Streptococcus vaccine compositions and methods of using the same |
| US20100316673A1 (en) * | 2009-06-16 | 2010-12-16 | The Regents Of The University Of Michigan | Nanoemulsion vaccines |
| DE102009031274A1 (en) * | 2009-06-30 | 2011-01-13 | Justus-Liebig-Universität Giessen | Liposomes for pulmonary application |
| CA2675704C (en) * | 2009-08-18 | 2010-05-25 | The Procter & Gamble Company | A method of making a fabric softening composition using a dynamic orifice with a valve in different positions |
| US10143652B2 (en) | 2009-09-23 | 2018-12-04 | Curirx Inc. | Methods for the preparation of liposomes |
| WO2011038073A1 (en) * | 2009-09-23 | 2011-03-31 | Formatech, Inc. | Methods for the preparation of liposomes comprising docetaxel |
| US8173589B2 (en) * | 2010-03-18 | 2012-05-08 | The Procter & Gamble Company | Low energy methods of making pearlescent fabric softener compositions |
| US9617323B2 (en) | 2010-06-07 | 2017-04-11 | Joshua Rabbani | Sulfonated sclerostin, antibodies, epitopes and methods for identification and use therefor |
| US9403882B2 (en) | 2010-06-07 | 2016-08-02 | Joshua Rabbani | Sulfation of Wnt pathway proteins |
| US9493541B2 (en) | 2010-06-07 | 2016-11-15 | Joshua Rabbani | Antibodies specific for sulfated sclerostin |
| WO2013006797A1 (en) | 2011-07-06 | 2013-01-10 | Nanobio Corporation | Human respiratory syncytial virus vaccine |
| JP6205360B2 (en) | 2011-08-22 | 2017-09-27 | ナノバイオ コーポレーション | Herpes simplex virus nanoemulsion vaccine |
| EP2753354A4 (en) | 2011-09-09 | 2015-04-15 | Nanobio Corp | Nanoemulsion respiratory syncytial virus (rsv) subunit vaccine |
| PT2773326T (en) | 2011-11-04 | 2019-04-23 | Nitto Denko Corp | Single use system for sterilely producing lipid-nucleic acid particles |
| US9579338B2 (en) | 2011-11-04 | 2017-02-28 | Nitto Denko Corporation | Method of producing lipid nanoparticles for drug delivery |
| CN104349783B (en) | 2012-05-21 | 2018-07-13 | 英斯麦德公司 | The system for treating pulmonary infection |
| RU2675859C2 (en) | 2012-11-29 | 2018-12-25 | Инсмед Инкорпорейтед | Stabilised vancomycin formulations |
| US9693958B2 (en) * | 2013-03-15 | 2017-07-04 | Cureport, Inc. | Methods and devices for preparation of lipid nanoparticles |
| US9868595B1 (en) * | 2013-05-20 | 2018-01-16 | James A. Scruggs | Vortex effect production device and method of improved transport of materials through a tube, pipe, and/or cylinder structure |
| GR1008481B (en) | 2013-12-05 | 2015-05-12 | Συμβουλοι Αναπτυξης Πωλησεων Επε, | Method for the confinement of plant oils (olive oil) with use of specific edible liposomes without phospholipids- application of said method in food, charcuterie, dairy products and fish preparations |
| PT3142643T (en) | 2014-05-15 | 2019-10-28 | Insmed Inc | Methods for treating pulmonary non-tuberculous mycobacterial infections |
| EP3204039B1 (en) | 2014-10-10 | 2022-06-08 | The Regents Of The University Of Michigan | Nanoemulsion compositions for preventing, suppressing or eliminating allergic and inflammatory disease |
| JP6316182B2 (en) | 2014-12-19 | 2018-04-25 | 富士フイルム株式会社 | Liposome production method and liposome production apparatus |
| CN107427465A (en) * | 2015-02-05 | 2017-12-01 | 马克·赛尔纳尔 | Ionic nano vesicle suspension and the biocide from its preparation |
| EP3270897A4 (en) | 2015-03-20 | 2018-12-05 | The Regents Of The University Of Michigan | Immunogenic compositions for use in vaccination against bordetella |
| US10675243B2 (en) | 2015-06-19 | 2020-06-09 | Global Health Solutions Llc | Petrolatum-based compositions comprising cationic biocides |
| US11173197B2 (en) | 2015-07-07 | 2021-11-16 | Bluewillow Biologics, Inc. | Methods and compositions for nanoemulsion vaccine formulations |
| WO2017201390A1 (en) | 2016-05-19 | 2017-11-23 | The Regents Of The University Of Michigan | Novel adjuvant compositions |
| EP3773505A4 (en) | 2018-03-30 | 2021-12-22 | Insmed Incorporated | Methods for continuous manufacture of liposomal drug products |
| US11369578B2 (en) | 2018-11-15 | 2022-06-28 | Bluewillow Biologics, Inc. | Persistent topical antimicrobial compositions and methods of using the same |
| CA3119836A1 (en) | 2018-11-15 | 2020-05-22 | Bluewillow Biologics, Inc. | Nanoemulsion compositions having enhanced permeability |
| AU2020241756A1 (en) | 2019-03-19 | 2021-11-04 | Arcturus Therapeutics, Inc. | Method of making lipid-encapsulated RNA nanoparticles |
| EP3711749A1 (en) * | 2019-03-19 | 2020-09-23 | Polymun Scientific Immunbiologische Forschung GmbH | Method of making lipid nanoparticles |
Family Cites Families (18)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US2043108A (en) * | 1930-04-22 | 1936-06-02 | Paul Lechler | Mixing apparatus for liquids |
| US3238021A (en) * | 1963-01-21 | 1966-03-01 | Monsanto Co | Mixing equipment |
| GB1193350A (en) * | 1967-11-09 | 1970-05-28 | Ilford Ltd | Liquid Mixing Apparatus |
| BE795321A (en) * | 1972-09-20 | 1973-08-13 | Gardinier Ets | REACTOR SUITABLE FOR REACTIONS BETWEEN TWO FLUIDS |
| SE387862B (en) * | 1974-09-13 | 1976-09-20 | G A Staaf | PIPE MIXER, INCLUDING A HOUSE DESIGNED AS A ROTARY BODY, TWO OR MORE CONNECTED PIPE PIPES FOR SUPPLYING THE MIXING COMPONENTS, AS WELL AS A TO THE HOUSE AXIALLY CONNECTED |
| FR2408387A2 (en) * | 1975-06-30 | 1979-06-08 | Oreal | COMPOSITIONS BASED ON AQUEOUS DISPERSIONS OF LIPID SPHERULES |
| US4019720A (en) * | 1975-10-16 | 1977-04-26 | Exxon Research And Engineering Company | Method and apparatus for mixing viscous materials |
| JPS5857970B2 (en) * | 1976-07-01 | 1983-12-22 | 高砂熱学工業株式会社 | Continuous dilution device for high viscosity fluids |
| DE2746188A1 (en) * | 1977-10-14 | 1979-04-19 | Bayer Ag | METHOD AND DEVICE FOR MIXING AND APPLYING REACTIVE SUBSTANCES |
| US4480925A (en) * | 1980-11-10 | 1984-11-06 | Dietrich David E | Method of mixing fluids |
| US4533254A (en) * | 1981-04-17 | 1985-08-06 | Biotechnology Development Corporation | Apparatus for forming emulsions |
| FR2534487B1 (en) * | 1982-10-15 | 1988-06-10 | Dior Christian Parfums | METHOD FOR THE HOMOGENEIZATION OF HYDRATED LIPIDAL LAMELLAR PHASE DISPERSIONS, AND SUSPENSIONS OBTAINED THEREBY |
| JPS607932A (en) * | 1983-06-29 | 1985-01-16 | Dai Ichi Seiyaku Co Ltd | Preparation of liposome |
| DE3442268C1 (en) * | 1984-03-09 | 1990-08-02 | Papierfabrik August Koehler AG, 7602 Oberkirch | Process for encapsulating dissolved reactants of color reaction systems, the capsules that are then available and their use in color reaction papers |
| US4753788A (en) * | 1985-01-31 | 1988-06-28 | Vestar Research Inc. | Method for preparing small vesicles using microemulsification |
| US4776991A (en) * | 1986-08-29 | 1988-10-11 | The United States Of America As Represented By The Secretary Of The Navy | Scaled-up production of liposome-encapsulated hemoglobin |
| US4911928A (en) * | 1987-03-13 | 1990-03-27 | Micro-Pak, Inc. | Paucilamellar lipid vesicles |
| JP2617346B2 (en) * | 1987-03-13 | 1997-06-04 | マイクロ ベシキュラー システムズ，インコーポレイテッド | Lipid vesicles formed from surfactants and steroids |
-
1988
- 1988-03-03 US US07/163,806 patent/US4895452A/en not_active Expired - Lifetime
-
1989
- 1989-03-02 BR BR898907293A patent/BR8907293A/en not_active IP Right Cessation
- 1989-03-02 WO PCT/US1989/000850 patent/WO1989007929A1/en active IP Right Grant
- 1989-03-02 AU AU32865/89A patent/AU614598B2/en not_active Ceased
- 1989-03-02 DE DE68910969T patent/DE68910969T2/en not_active Expired - Fee Related
- 1989-03-02 EP EP89903640A patent/EP0406273B1/en not_active Expired - Lifetime
- 1989-03-02 JP JP1503339A patent/JP2542271B2/en not_active Expired - Fee Related
- 1989-03-02 KR KR1019890702024A patent/KR900700072A/en not_active Application Discontinuation
- 1989-03-03 CA CA000592763A patent/CA1320400C/en not_active Expired - Lifetime
Also Published As
| Publication number | Publication date |
|---|---|
| EP0406273A1 (en) | 1991-01-09 |
| EP0406273A4 (en) | 1991-08-28 |
| KR900700072A (en) | 1990-08-11 |
| DE68910969D1 (en) | 1994-01-05 |
| AU614598B2 (en) | 1991-09-05 |
| JP2542271B2 (en) | 1996-10-09 |
| BR8907293A (en) | 1991-03-12 |
| AU3286589A (en) | 1989-09-22 |
| JPH03504101A (en) | 1991-09-12 |
| WO1989007929A1 (en) | 1989-09-08 |
| EP0406273B1 (en) | 1993-11-24 |
| DE68910969T2 (en) | 1994-06-23 |
| US4895452A (en) | 1990-01-23 |
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| MKEX | Expiry |
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