CA2211262C - Micellar nanoparticles - Google Patents
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- CA2211262C CA2211262C CA002211262A CA2211262A CA2211262C CA 2211262 C CA2211262 C CA 2211262C CA 002211262 A CA002211262 A CA 002211262A CA 2211262 A CA2211262 A CA 2211262A CA 2211262 C CA2211262 C CA 2211262C
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- 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
- A61K9/107—Emulsions ; Emulsion preconcentrates; Micelles
- A61K9/1075—Microemulsions or submicron emulsions; Preconcentrates or solids thereof; Micelles, e.g. made of phospholipids or block copolymers
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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- Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10S977/00—Nanotechnology
- Y10S977/70—Nanostructure
- Y10S977/773—Nanoparticle, i.e. structure having three dimensions of 100 nm or less
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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- Y10S977/902—Specified use of nanostructure
- Y10S977/904—Specified use of nanostructure for medical, immunological, body treatment, or diagnosis
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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- Y10S977/00—Nanotechnology
- Y10S977/902—Specified use of nanostructure
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Abstract
The present invention relates to micellar nanoparticles and methods of their production. Micellar nanoparticles are made by hydrating a mixture of an oil, a stabilizer/surfactant, and an alcoholic initiator with an aqueous solution. These micellar nanoparticles are normally less than 100 nanometers in diameter. The micellar nanoparticles are particularly advantageous in delivering materials such as estradiol topically through the skin because their small size allows easy penetration.
Description
MICELLAR NANOPARTICLES
Background of the Invention _ __ The present invention is concerned with the materials and methods for constructing "micellar nanoparticles." micelle-like particles with mean diameters less than nanometers (one micron). These micellar nanoparticles are submicron-sized, oil-based particles. the smallest of which are filterable through a 0.2 micron filter such as is standardly used for microbiological purification. The micellar nanoparticles of the invention may be formed into stable dispersions in aqueous solutions and buffers.
The micellar nanoparticles have a variety of uses because of their small size.
Other synthetic particles such as liposomes, nonphospholipid lipid vesicles and microcapsules are normally a micron or larger. In contrast, it is possible to form the micellar nanoparticles of the invention in sizes less than 100 nanometers diameter. Unlike lipid vesicles. some of which can be engineered to carry an oil, see, e.g., United States Patent No.
4, 911,928 to Wallach. the present particles reauire at least an oil. a stabilizer/surfactant. an initiator. and water or another diluent in their manufacture. However, neither cholesterol nor phospholipids are used. In fact, these nanoparticles can be made using food grade, USP or NF grade materials suitable for human use applications. This is particularly important if these micellar nanoparticles are to be used for topical delivery of a material into the bloodstream. One specific use of this type of system is the delivery of natural or synthetic hormones such as estradiol. These materials often have solubility problems;
e.g., they are often only soluble in materials such as ethanol which can be difficult to incorporate in stable particulate systems.
Micellar nanoparticles are unique in that they allow materials that are soluble in any of water. oil, or the initiator (i.e., ethanol or methanol) to be incorporated into stable particles with mean diameters between about 30 and 1000 nanometers. Most preparations have particle diameters between 30 to 500 nanometers, are mixable in water. and filterable through either 0.2 or 0.4~ micron filters. They can be stored at between -20 and 25 degrees C°.
Utilizing the materials and methods describe, one can produce micellar nanoparticles that do the following:
1. Incorporate ethanol or methanol soluble drugs into the particles.
Background of the Invention _ __ The present invention is concerned with the materials and methods for constructing "micellar nanoparticles." micelle-like particles with mean diameters less than nanometers (one micron). These micellar nanoparticles are submicron-sized, oil-based particles. the smallest of which are filterable through a 0.2 micron filter such as is standardly used for microbiological purification. The micellar nanoparticles of the invention may be formed into stable dispersions in aqueous solutions and buffers.
The micellar nanoparticles have a variety of uses because of their small size.
Other synthetic particles such as liposomes, nonphospholipid lipid vesicles and microcapsules are normally a micron or larger. In contrast, it is possible to form the micellar nanoparticles of the invention in sizes less than 100 nanometers diameter. Unlike lipid vesicles. some of which can be engineered to carry an oil, see, e.g., United States Patent No.
4, 911,928 to Wallach. the present particles reauire at least an oil. a stabilizer/surfactant. an initiator. and water or another diluent in their manufacture. However, neither cholesterol nor phospholipids are used. In fact, these nanoparticles can be made using food grade, USP or NF grade materials suitable for human use applications. This is particularly important if these micellar nanoparticles are to be used for topical delivery of a material into the bloodstream. One specific use of this type of system is the delivery of natural or synthetic hormones such as estradiol. These materials often have solubility problems;
e.g., they are often only soluble in materials such as ethanol which can be difficult to incorporate in stable particulate systems.
Micellar nanoparticles are unique in that they allow materials that are soluble in any of water. oil, or the initiator (i.e., ethanol or methanol) to be incorporated into stable particles with mean diameters between about 30 and 1000 nanometers. Most preparations have particle diameters between 30 to 500 nanometers, are mixable in water. and filterable through either 0.2 or 0.4~ micron filters. They can be stored at between -20 and 25 degrees C°.
Utilizing the materials and methods describe, one can produce micellar nanoparticles that do the following:
1. Incorporate ethanol or methanol soluble drugs into the particles.
2. Incorporate ethanol or methanol soluble pesticides into the particles.
3. Incorporate adjuvants into the particles.
4. Incorporate proteins into the particles.
5. Incorporate whole viruses containing intact nucleic acids-into the particles. It must be noted. however, that the smaller particles of the invention are about the same size as many viruses.
6. Incorporate ethanol-extracted flavors into the particles.
7. Incorporate volatile oils (flavors and fragrances) into the particles.
8. Incorporate a charge into the particles.
9. Create colored particles.
Of particular importance is the ability to transmit drugs topically. It has been known for many years that small particles, such as those below one micron in diameter. can more easily traverse the skin boundary than larger particles. However, the small amount of drug transmitted in small particles has often limited their usefulness In addition, most particles have only had limited classes of materials they could deliver.
Accordingly, an object of the invention is to produce submicron particles which can deliver a variety of classes of materials.
Another object of the invention is to produce submicron particles that can deliver materials that are soluble in ethanol or methanol but have limited or no solubility in aqueous and oil systems.
A further object of the invention is to produce particles below 100 nanometers in diameter that can be used for drug delivery.
A still further object of the invention is to produce a particle for topical delivery of hormones such as estradiol.
These and other objects and features of the invention will be apparent from the description and the claims.
Summary of the Invention The present invention features micellar nanoparticles and methods of their manufacture. These micellar nanoparticles have particular utility as drug delivery vehicles with specific applications to topical delivery of materials that are soluble in ethanol and methanol. However, these micellar nanoparticles can also be used to deliver many different classes of drugs and other materials. The small size of the micellar nanoparticles and their compatibility with tissue render them applicable to numerous uses.
The micellar nanoparticles of the invention have diameters of about 10-1000 nanometers, with most of the particles having diameters of under I 00 nanometers. This small particle size allows passage through a 0.2 micron filter. The nanoparticles are made of a lipophilic phase which includes an oil, a stabilizer (or surfactant) and an initiator such as ethanol or methanol. This lipophilic phase is hydrated by an aqueous solution such as water or a buffer. Preferred stabilizers are non-phospholipid surfactants, particularly the Tween* (polyoxyethylene derivatives of sorbitan fatty acid esters) family of surfactants and the nonylphenol polyethylene glycol ethers. Most preferred surfactants are Tween*
60 (polyoxyethylene 20 sorbitan monostearate) and Tween* 80 (polyoxyethylene sorbitan monooleate), and Tergitol* NP-40 (Poly(oxy-1,2-ethanediyl), a-(4-nonylphenol)-c~-hydroxy, branched [molecular weight average 1980]) and Tergitol* NP-70 (a mixed surfactant-AQ=70%). The high molecular weight of these surfactants appears to have advantageous properties in manufacture and stability of the resulting micellar nanoparticles.
The preferred initiators in the present invention are ethanol and methanol, but other short chain alcohols and/or amides may be used in certain circumstances. While pure ethanol or methanol are preferred, mixtures of the two, and materials, blended or unblended, containing at least 50% alcohol, can be used. This group of initiators can *Trade Mark include flavored initiators such as alcoholic extracts of flavors like peppermint, lemon, orange and the like.
In addition to the initiator and the surfactant of stabilizer, the micellar particles can be modified or custom manufactured by selection of the proper oil. While most oils seem to work, the preferred oils are selected from the group consisting of vegetable oils, nut oils, fish oils, lard oil, mineral oils, squalane, tricaprylin, and mixtures thereof.
A number of other materials may be added to the micellar nanoparticles to customize the particles. Volatile oils, such as volatile flavor oils, can be used in lieu of some of the oil or can be added in addition to the other oil forming the particles. A
coloring agent, such as a food coloring agent can also be used, preferably by adding it to the initiator. The initiator or the oil can also carry actives which are incorporated into the final particle suspension. These actives can be dissolved, or suspended in the liquid. One preferred additive is a steroid or hormone such as estradiol, which can be dissolved in an ethanol initiator and incorporated into the particle. Since estradiol precipitates in aqueous solutions, the addition of the aqueous phase will precipitate the estradiol, which can then be released in a topical preparation. One interesting fact that appears is that the type of crystals formed using the methods of the present invention are different in shape than standard aqueous solution precipitates of estradiol.
The aqueous solution which is used to hydrate the lipophilic phase is preferably a physiologically compatible solution such as water or a buffer, e.g., phosphate buffered saline. The aqueous solution may have an active material dissolved or suspended therein for incorporation. The basic procedure for the manufacture of the micellar nanoparticles is blending the oil, the stabilizer/surfactant, and the initiator to form a lipophilic phase and blending an excess, preferably about a 4:1 ratio, of the lipophilic phase with an aqueous dilulent solution. The blending, or hydrating, of the lipophilic phase with the aqueous phase is preferably accomplished using a device which generates a relative velocity of about 50 m/s through an orifice diameter of 1/18,000 of an inch.
This shear -4a-provides particles in the preferred size range while lower shear values, e.g., by using larger orifices or lower velocities, can cause larger particle size.
All of the different materials and processes described herein can be modified or selected to control the properties of the resulting micellar nanoparticles.
Actives can be carried in the oil, the initiator, or the aqueous phase for incorporation into the particles.
Although it appears that the particles are micelles, they may be in the form of reverse micelles without changing the scope of the invention. The invention is further illustrated by the following detailed description and the drawing.
An aspect of the present invention provides a micellar nanoparticle having a diameter of between about 25 and 1000 nm, said micellar nanoparticle comprising a lipophilic phase which includes an oil, a stabilizer and an alcohol-based initiator, hydrated with a suitable aqueous-based solution wherein said stabilizer is selected from the group consisting of Tween* 60, Tween* 80, nonylphenol polyethylene glycol ethers, and mixtures thereof.
Another aspect of the present invention provides a method of making micellar nanoparticles comprising the steps of: blending an excess of an oil, together with a stabilizer and an initiator to form a lipophilic phase, said stabilizer being selected from the group consisting of Tween* 60, Tween* 80, Nonylphenol Polyethylene Glycol Ethers, and mixtures thereof; preparing a dilulent solution having an aqueous solution base; and blending an excess of said lipophilic phase with said dilulent to form said micellar nanoparticles.
Brief Description of the Drawing Figures la and lb are electromicrographs of the nanoparticles of the invention at two different magnifications; and *Trade Mark -4b-Figure 2 is a graph of serum estradiol lelvels in ovariectomized Rhesus monkeys following topical administration of 1 mg of estradiol using three different types of vehicles.
Detailed Description of the Invention The present invention concerns micellar nanoparticles and methods of their production. Unlike microcapsules and liposomal systems, the present micellar nanoparticles have a significant size population under 100 nanometers in diameter. while still carrying significant quantities of active ingredients. These micellar nanoparticles are particularly useful as topical drug delivery vehicles because their small size and other characteristics which permit rapid dermal penetration. The micellar nanoparticles are also exceptionally versatile in that the active materials which can be carried include those which are suspendable or dissolvable in any of the oil. aqueous dilulent, or, preferable. the initiator. These properties allow this system to be used with actives that are difficult to use in other delivery systems.
Micellar nanoparticles are formed by first combining at least one oil, preferably an oil selected from Table 1. a stabilizer (surfactant), preferably a surfactant from Table 2. and an initiator. preferably ethanol or methanol. Most preferred stabilizers are Tweerf60. Tweeri'80.
Tergitol~lP-40 and Tergito~NP-70. Additional possible initiators are shown in Table 3 (alcohols and related compounds) and Table 4 (alcohol flavored extracts). If any of the 1 ~ alcohol flavored extracts of Table 4 are used which are less than ~0%
ethanol, a 1:1 mixture of ethanol and the extract is used to ensure that at least 50% ethanol is used. Volatile oils can also be added to these chemical components (Table 5), and colors may also be added to the oil-stabilizer-initiator mixture (Table 6). A negative charge may be introduced by addition of oleic acid to the oil-stabilizer-initiator mixture. After pre-mixing these materials, water or a suitable buffer such as those shown in Table 7 is injected under a high velocity into this mixture. The preferred ratio of oilatabilizer:initiator is 25:3:5, respectively, on a volume per volume basis. The preferred ratio of the pre-mixed oil containing phase to water is 4:1, respectively. Nanoparticles can be produced with reciprocating syringe instrumentation, continuous flow instrumentation, or high speed mixing equipment. Particles created at this 4:1 ratio range in diameters from 30 to 500 nanometers. These water miscible particles can then be filtered through either a 0.2 or 0.45 micron filter. Larger micellar particles can be created by simply increasing the water content. decreasing the oil-stabilizer-initiator content.
or changing the shear in forming the particles. We have coined the name "micellar nanoparticles" for particles with mean diameters less than 1000 nanometers (one micron).
'Trade Mark TABLE 1: Oils Utilized in Preparation of Micellar Nanoparticles .
Almond oil, sweet Apricot seed oil Borage oil Canola oil Coconut oil Corn oil Cotton seed oil Fish oil Jojoba bean oil Lard oil Linseed oil, boiled Macadamia nut oil Mineral oil Olive oil Peanut oil Safflower oil Sesame oil Soybean oil Squalane Sunflower seed oil Tricaprylin (1, ?, 3 trioctanoyl glycerol) Wheat germ oil -TABLE 2: Stabilizers/Surfactants Utilized in Preparation of Micellar Nanoparticles.
Tween#60 Tween#80 Nonvlphenol Polyethylene Glycol Ethers (alkylphenol-hydroxypolyoxyethylene) 1. Poly(oxy-1. 2-ethanedivl), alpha-(4-nonylphenol)-omega-hv_ droy-, branched (i.e. TergitoI~NP-40 Surfactant) Formula: C95H 1 g~040 MW (average) = 1980 1 ~ '_'. Nonylphenol Polyethylene Glycol Ether mixtures (i.e. TergitoI~NP-70 (70%AQ) Surfactant]
Formula and MV: not applicable (mixture) ?0 TABLE 3: Initiators Utilized in Preparation of Micellar Nanoparticles.
Ethanol Methanol 3~Trade Mark WO 96/23409 PCTlUS96/01410 _g_ TABLE 4: Flavored Initiators (flavored extracts*) Utilized in Preparation o f Micellar Nanoparticles.
Pure Anise extract (73% Ethanol) Imitation Banana extract (40% Ethanol) Imitation Chem~ extract (24% Ethanol) Chocolate extract (23% Ethanol) Pure Lemon extract (84% Ethanol) Pure Orange extract (80% Ethanol) Pure Peppermint extract (89% Ethanol) Imitation Pineapple extract (42% Ethanol) Imitation Rum extract (35% Ethanol) Imitation Strawberry extract (30% Ethanol) Pure Vanilla extract (35% Ethanol) * Extracts utilized are food grade materials (McCormick). Materials from other sources could be substituted.
TABLE 5: Volatile Oils or Fragrances Utilized in Preparation of Micellar Nanoparticles.
Balm oil Bay oil Bergamot oil Cedarwood oil Cherry oil Cinnamon oil 4~
Clove oil Origanum oil Peppermint oil WO 96/23409 PCTlUS96101410 -S-TABLE 6: Food Colors Utilized in Preparation of Micellar Nanoparticles.
Green Yellow Red Blue * Food colors utilized are food grade materials (McCormick). Materials from other sources could be substituted.
TABLE 7: List of Diluents Utilized in Preparation of Micellar Nanoparticles.
Water for injection Phosphate buffered saline The following Examples will more clearly illustrate the invention and its usefulness.
ExamQle 1- Production of Uncharged Micellar Nanooarticles Table 8 contains the materials used to produce micellar nanoparticles where water is the diluent. Sizing parameters using a Coulter L 130 Laser sizing apparatus are shown in Table 9.
TABLE 8: Preparation of Micellar nanoparticles utilizing water as the diluent.
Chemical Component Amount Soybean oil (Oil) '_'~ mL
Polysorbate 80 (Tween~80) (Stabilizer) 3 mL
Ethanol (Initiator) s mL
The above Oil-Stabilizer-Initiator components are mixed for 60 seconds. One mL
of water is injected into four mL of the mixture using reciprocating syringe instrumentation. This instrumentation has two ~mL syringes connected together through a stainless steel Leurlok connector with a 1/18,000 inch orifice. The solutions are driven between the syringes.
through the connector, for about 100 seconds. The resulting particles were dried on EM
grids, stained with uranyl acetate. and electron micrograph studies performed.
Figure 1 a shows an efectromicrograph of this preparation at a 60.OOOX magnification while Figure 1 b shows the same preparation at a 1 ~O,OOOX magnification. A brief description of the method of production of the micellar nanoparticles follows each table.
Table 9 - Sizing of Micellar Nanoparticles using water as a Diluent Mean Diameter Range Preparation ~nanometersl_ (nanomete~l_ Micellar nanoparticles 312 193-4s~
(SBOIT'v~0/E/WFI) One problem with using the LS 130 sizing device is that it cannot accurately size particles which are less than 200 nanometers in diameter. Using Figures 1 a and 1 b, it is determined that most of the particles are between 70 and 90 nanometers in diameter. with only ~% of particles be greater than 90 nanometers in diameter. Particles in the range of 20-30 nanometers are visible in the higher magnification shown in Figure lb.
3ETrade Mark Example 2 - Incorporation of Estradiot into Micellar Nanoparticles Tables 10 and 12 contain the materials utilized to produce two lots of uncharged a micellar nanoparticles into which estradiol has been incorporated at two different concentrations. Both preparations are made using water as the diluent. The higher estradiol concentration materials were used in the rhesus monkey studies described in Example 3 below . Either ~0 or 100 mg of estradiol is solublized in the initiator (ethanol component) of the preparation prior to formation of the micellar nanoparticles. This is necessar~~ since estradiol precipitates in the presence of water. In fact, the small amount of water in the reagent grade ethanol appears to be sufficient to precipitate the estradiol since the micellar particles formed using the materials and procedures described herein appear to have crystals of estradiol contained therein. However, these crystals appear to have a sheet-like form rather than the needle-like form standardly found in water precipitation.
TABLE 10: Preparation of Micellar Nanoparticles Containing Estradiol Soybean oil (Oil) 25mL
Polysorbate 80 (Tween 80) (Stabilizer) 3mL
Ethanol (Initiator) SmL
Estradiol SOmg The micellar nanoparticles were made using procedures substantially identical to that described in Example 1, except the estradiol was dissolved (or suspended) in the ethanol initiator prior to the mixing of the initiator with the other components. The oil-stabilizer-initiator/estradiol components are hand mixed or can be mixed for 60 seconds using a vortex mixer. One mL of water is injected into four mL of the resulting mixture using reciprocating syringe instrumentation such as is described in Example 1.
w TABLE 11 - Sizing data on Estradiol containing Micellar Nanoparticles (50 mg) Mean Diameter Range Preparation (nanometers) (nanometers) Micellar nanoparticles 289 174-459 (SBO/Tw#80/Etoh-estradiol/WFI) Sizing data on these preparations, measured using a Coulter LS130 Laser sizing apparatus. is shown in Tables 1 l and 13, respectively, for the two preparations.
The LS130 sizing device cannot size particles accurately less than 200 nanometers in diameter. These materials were also dried on EM grids, stained with uranyl acetate and electron micrograph studies performed. Electron micrographs reveal that most of the particles are less than 200 nanometers. Particles in the range of 20-30 nanometers are visible.
Crystallized estradiol is readily visible in the larger micelles. No free drug crystals are noted in any fields suggesting complete incorporation of drug into micelles.
TABLE 12: Preparation of Micellar Nanoparticles Containing Estradiol Chemical Component Amount Soybean oil (Oil) ?~mL
Polysorbate 80 (Tween~90) (Stabilizer) 3mL
Ethanol (Initiator) ~mL
Estradiol 100mg #Trade Mark ' -13-TABLE 13 -Sizing data on Estradiol containing Micellar Nanoparticles (100 mg) Mean Diameter Range Preparation ~nanometersl ~.anometersl Micellar nanoparticles 217 1 ~ 1-291 (SBO/Tw~O/Etoh-estradiol/WFI) Example 3 - Rhesus Monkey Testing of Estradiol Containing Preparations The 100 mg estradiol preparation of Example two was tested against a standard ethanol preparation of estradiol to show efficacy. One milligram of estradioh in either ethanol (Table 14) or micellar nanoparticles (Table 15), was applied to the skin of groups of four ovariectomized rhesus monkeys. Serial blood samples were drawn and serum estradiol levels were determined over the next 32 days. The serum estradiol data is graphically depicted in Figure ?. No additional drug was applied to skin of any animal.
Animals were observed for the next 60 davs to determine whether the time of occurrence.
duration and severity of vaginal bleeding (Table 16).
#Trade Mark WO 96/23409 PCT/tJS96/01410 TABLE 14 -Serum Estradiol Levels in Ovariectomized Female Monkeys Following a Single Topical Application of Micellar Nanoparticles Equivalent to 1 mg Estradiol Monkey Number k.
Sample Serum stradiol Group Mean E
Time #19567 #21792 #22366 #22405 S.E.
(pg/ml) (pg/ml)(pg/ml)(pg/ml) 0 hour 0.0b 0.0b 0.0b 0.0b 0.0 0.0h 0.5 hour 22.2 49.8 36.9 77.5 46.6 11.7 1 hour 37.4 60.9 65.6 108.6 68.1 14.8 2 hours 61.5 80.5 87.3 191.3 105.2 29.2 4 hours 77.2 132.1 120.6 120.4 112.6 12.1 6 hours 89.0 166.3 119.0 158.3 133.2 18.0 8 hours 87.5 157.3 116.1 148.1 127.3 15.9 12 hours 83.0 160.5 100.6 140.3 121.1 17.8 day 1 90.7 178.0 105.7 132.6 126.8 19.2 day 2 95.5 152.8 90.6 83.5 105.6 15.9 day 3 81.9 122.6 51.1 47.2 75.7 17.5 day 4 91.5 83.9 58.7 50.3 71.1 9.9 day ~ 41.6 74.7 3 5.1 40.0 47.9 9.1 day 6 45.2 63.7 25.6 40.9 43.9 7.8 day 7 18.3 25.9 21.9 27.0 23.3 2.0 daV 12 0.0b 0.0b 0.0b 0.0b 0.0 0.06 daV 17 0.0b 0.0b 0.0b 0.06 0.0 0.0b daV 22 0.0b 0.0b 0.0b 0.0b 0.0 0.06 daV 27 0.0b 0.0b 0.0b 0.0b 0.0 0.0b day 32 0.0b 0.0b 0.0b 0.0b 0.0 -~' 0.0b a CDB 3988 = 2.4 mg estradiol/ml of Tween/Oil. The dosing volume was 0.42 ml.
b 0 = Not Detectable. The limit of detection (ED9o) for the assay was 13.3 ~ 2.4 pg/ml (mean ~ S.E., n = 4) TABLE 15 - Serum Estradiol Levels in Ovariectomized Female Monkeys Following a Single Topical Application of 1 mg Ethanol Containing Estradiol$
Monkev Number Sample Serum Group Mean Estradiol Time #G-558 #G-603 #E-920 #E-924 S.E.
(pg/ml~(pg/ml) (pg/ml)(pg/ml) 0 hour 0.0b 0.06 0.0b 0.0b 0.0 0.06 0.5 hour 17.7 97.1 44.8 19.5 44.8 18.5 1 hour 53.2 44.1 88.3 99.9 71.4 13.5 2 hours 144.3 89.4 138.5 155.1 131.8 14.6 4 hours 143.7 202.3 165.1 193.6 176.2 13.4 6 hours 155.8 257.8 173.1 203.7 197.6 22.4 I
8 hours 114.2 266.1 130.7 130.0 160.3 35.5 12 hours 80.8 219.5 86.4 115.9 125.7 32.2 day 1 92.4 145.2 56.9 109.4 101.0 18.4 day 2 74.1 124.2 55.3 107.2 90.2 15.6 day 3 65.0 67.4 51.9 89.2 68.4 7.7 day 4 70.5 79.6 57.8 90.0 74.5 6.8 day 5 53.6 53.2 51.6 47.3 51.4 1.4 day 6 60.1 59.0 59.4 53.0 57.9 1.6 day 7 48.7 40.6 50.3 36.6 44.1 3.3 day 12 28.5 24.2 53.3 O.Ob 26.4 10.9b day 17 O.Ob O.Ob 28.9 O.Ob 7.2 7.2b day 22 O.Ob O.Ob 13.8 0.06 3.5 3.5 I, day 27 O.Ob 0.0b 0.0b 0.06 0.0 0.0b I'' day 32 0.06 0.0b 0.0b 0.0b 0.0 0 .0b a CDB 100 = 2.4 mg estradiol/ml of absolute ethanol. The dosing volume was 0.42 ml.
b 0 = Not Detectable. The limit of detection (ED9o) for the assay was 13.3 ~ 2.4 pg/ml (mean ~ S.E., n = 4) The data in Tables 14 and 1 ~ and Figure 2 show that therapeutic serum levels of estrogen are present in the blood stream of ovariectomized animals in both groups in one t hour after a single application. Mean estradiol levels greater than 40 picograms/ml are maintained for 7 days with the ethanol preparation and for 6 days with the nanoparticle r preparation. When estrogen levels remain low (see Figure 2 and Table 16), vaginal bleeding occurs in both groups. Also of particular interest is the shape of the curves in Figure 2. The ethanol-estradiol preparation yields a "shark tooth" curve showing a high initial action and a sharp fall-off while the micellar nanoparticle preparation yields more of a "mesa" effect with a nearly flat level for several hours. This "mesa" effect is often preferred since some of the problems associated with peaking can be minimized.
RHESUS MONKEYS FOLLOWING A SINGLE TOPICAL APPLICATION OF
ESTRADIOL IN ALCOHOL OR
MICELLAR NANOPARTICLES
WITHDRAWAL EDING
BLE
CDB No. ESTRADIOL ESTER DAYS INTENSITYa ' LATENCY DURATION
100 Estradiol in alcoholic19.5 0.3 4.3 0.9 1.6 0.2 solution 3988 Estradiol formulationb16.5 0.5~7.3 1.5 1.6 0.1 aMean intensity of bleeding (1=scant, moderate, 3=heavy) over bleeding period bNovavax MN Suspension 11294-2 Significantly different (p<0.01) from estradiol in alcohol solution based on a one-way analysis of variance followed by a Student Neuman-Keuls multiple range test Therefore, this Example demonstrates in a non-human primate that the micellar nanoparticles of the invention can be utilized to deliver estradiol through intact skin with maintenance of therapeutic serum estradiol levels for 6 days after a single application. This technology may have numerous therapeutic applications in medicine.
., 5 The estadiol preparation is also stable at a variety of temperatures. Table 17 shows thermal stability data for the micellar nanoparticle preparation of the Example 2 at -20°C, 25°C, and 65°C. As is clear, while the micellar nanoparticles are unstable at high temperatures, they are stable at room temperature and low temperatures.
TABLE 17: Thermal Stability of Micellar Nanoparticles Preparation Mean Range Diameter (nanometers) (nanometers) Micellar nanoparticles 361 168-599 (SBO/Tw80/Etoh-estradiol/WFI) Storage at 25°C
Micellar nanoparticles 312 179-510 (SBO/Tw80/Etoh-estradiol/WFI) Storage at -20°C
Micellar nanoparticles Unstable (SBO/Tw80/Etoh-estradiol/WFI) Storage at 65°C
In addition, the micellar nanoparticles of the invention can be diluted with aqueous solutions without stability loss. This allows the possibility of using high concentration products which can be diluted for use as necessary.
Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation. many equivalents to the specific embodiments of the invention described herein. Such equivalents are intended to be encompassed by the following claims.
a What is claimed is:
Of particular importance is the ability to transmit drugs topically. It has been known for many years that small particles, such as those below one micron in diameter. can more easily traverse the skin boundary than larger particles. However, the small amount of drug transmitted in small particles has often limited their usefulness In addition, most particles have only had limited classes of materials they could deliver.
Accordingly, an object of the invention is to produce submicron particles which can deliver a variety of classes of materials.
Another object of the invention is to produce submicron particles that can deliver materials that are soluble in ethanol or methanol but have limited or no solubility in aqueous and oil systems.
A further object of the invention is to produce particles below 100 nanometers in diameter that can be used for drug delivery.
A still further object of the invention is to produce a particle for topical delivery of hormones such as estradiol.
These and other objects and features of the invention will be apparent from the description and the claims.
Summary of the Invention The present invention features micellar nanoparticles and methods of their manufacture. These micellar nanoparticles have particular utility as drug delivery vehicles with specific applications to topical delivery of materials that are soluble in ethanol and methanol. However, these micellar nanoparticles can also be used to deliver many different classes of drugs and other materials. The small size of the micellar nanoparticles and their compatibility with tissue render them applicable to numerous uses.
The micellar nanoparticles of the invention have diameters of about 10-1000 nanometers, with most of the particles having diameters of under I 00 nanometers. This small particle size allows passage through a 0.2 micron filter. The nanoparticles are made of a lipophilic phase which includes an oil, a stabilizer (or surfactant) and an initiator such as ethanol or methanol. This lipophilic phase is hydrated by an aqueous solution such as water or a buffer. Preferred stabilizers are non-phospholipid surfactants, particularly the Tween* (polyoxyethylene derivatives of sorbitan fatty acid esters) family of surfactants and the nonylphenol polyethylene glycol ethers. Most preferred surfactants are Tween*
60 (polyoxyethylene 20 sorbitan monostearate) and Tween* 80 (polyoxyethylene sorbitan monooleate), and Tergitol* NP-40 (Poly(oxy-1,2-ethanediyl), a-(4-nonylphenol)-c~-hydroxy, branched [molecular weight average 1980]) and Tergitol* NP-70 (a mixed surfactant-AQ=70%). The high molecular weight of these surfactants appears to have advantageous properties in manufacture and stability of the resulting micellar nanoparticles.
The preferred initiators in the present invention are ethanol and methanol, but other short chain alcohols and/or amides may be used in certain circumstances. While pure ethanol or methanol are preferred, mixtures of the two, and materials, blended or unblended, containing at least 50% alcohol, can be used. This group of initiators can *Trade Mark include flavored initiators such as alcoholic extracts of flavors like peppermint, lemon, orange and the like.
In addition to the initiator and the surfactant of stabilizer, the micellar particles can be modified or custom manufactured by selection of the proper oil. While most oils seem to work, the preferred oils are selected from the group consisting of vegetable oils, nut oils, fish oils, lard oil, mineral oils, squalane, tricaprylin, and mixtures thereof.
A number of other materials may be added to the micellar nanoparticles to customize the particles. Volatile oils, such as volatile flavor oils, can be used in lieu of some of the oil or can be added in addition to the other oil forming the particles. A
coloring agent, such as a food coloring agent can also be used, preferably by adding it to the initiator. The initiator or the oil can also carry actives which are incorporated into the final particle suspension. These actives can be dissolved, or suspended in the liquid. One preferred additive is a steroid or hormone such as estradiol, which can be dissolved in an ethanol initiator and incorporated into the particle. Since estradiol precipitates in aqueous solutions, the addition of the aqueous phase will precipitate the estradiol, which can then be released in a topical preparation. One interesting fact that appears is that the type of crystals formed using the methods of the present invention are different in shape than standard aqueous solution precipitates of estradiol.
The aqueous solution which is used to hydrate the lipophilic phase is preferably a physiologically compatible solution such as water or a buffer, e.g., phosphate buffered saline. The aqueous solution may have an active material dissolved or suspended therein for incorporation. The basic procedure for the manufacture of the micellar nanoparticles is blending the oil, the stabilizer/surfactant, and the initiator to form a lipophilic phase and blending an excess, preferably about a 4:1 ratio, of the lipophilic phase with an aqueous dilulent solution. The blending, or hydrating, of the lipophilic phase with the aqueous phase is preferably accomplished using a device which generates a relative velocity of about 50 m/s through an orifice diameter of 1/18,000 of an inch.
This shear -4a-provides particles in the preferred size range while lower shear values, e.g., by using larger orifices or lower velocities, can cause larger particle size.
All of the different materials and processes described herein can be modified or selected to control the properties of the resulting micellar nanoparticles.
Actives can be carried in the oil, the initiator, or the aqueous phase for incorporation into the particles.
Although it appears that the particles are micelles, they may be in the form of reverse micelles without changing the scope of the invention. The invention is further illustrated by the following detailed description and the drawing.
An aspect of the present invention provides a micellar nanoparticle having a diameter of between about 25 and 1000 nm, said micellar nanoparticle comprising a lipophilic phase which includes an oil, a stabilizer and an alcohol-based initiator, hydrated with a suitable aqueous-based solution wherein said stabilizer is selected from the group consisting of Tween* 60, Tween* 80, nonylphenol polyethylene glycol ethers, and mixtures thereof.
Another aspect of the present invention provides a method of making micellar nanoparticles comprising the steps of: blending an excess of an oil, together with a stabilizer and an initiator to form a lipophilic phase, said stabilizer being selected from the group consisting of Tween* 60, Tween* 80, Nonylphenol Polyethylene Glycol Ethers, and mixtures thereof; preparing a dilulent solution having an aqueous solution base; and blending an excess of said lipophilic phase with said dilulent to form said micellar nanoparticles.
Brief Description of the Drawing Figures la and lb are electromicrographs of the nanoparticles of the invention at two different magnifications; and *Trade Mark -4b-Figure 2 is a graph of serum estradiol lelvels in ovariectomized Rhesus monkeys following topical administration of 1 mg of estradiol using three different types of vehicles.
Detailed Description of the Invention The present invention concerns micellar nanoparticles and methods of their production. Unlike microcapsules and liposomal systems, the present micellar nanoparticles have a significant size population under 100 nanometers in diameter. while still carrying significant quantities of active ingredients. These micellar nanoparticles are particularly useful as topical drug delivery vehicles because their small size and other characteristics which permit rapid dermal penetration. The micellar nanoparticles are also exceptionally versatile in that the active materials which can be carried include those which are suspendable or dissolvable in any of the oil. aqueous dilulent, or, preferable. the initiator. These properties allow this system to be used with actives that are difficult to use in other delivery systems.
Micellar nanoparticles are formed by first combining at least one oil, preferably an oil selected from Table 1. a stabilizer (surfactant), preferably a surfactant from Table 2. and an initiator. preferably ethanol or methanol. Most preferred stabilizers are Tweerf60. Tweeri'80.
Tergitol~lP-40 and Tergito~NP-70. Additional possible initiators are shown in Table 3 (alcohols and related compounds) and Table 4 (alcohol flavored extracts). If any of the 1 ~ alcohol flavored extracts of Table 4 are used which are less than ~0%
ethanol, a 1:1 mixture of ethanol and the extract is used to ensure that at least 50% ethanol is used. Volatile oils can also be added to these chemical components (Table 5), and colors may also be added to the oil-stabilizer-initiator mixture (Table 6). A negative charge may be introduced by addition of oleic acid to the oil-stabilizer-initiator mixture. After pre-mixing these materials, water or a suitable buffer such as those shown in Table 7 is injected under a high velocity into this mixture. The preferred ratio of oilatabilizer:initiator is 25:3:5, respectively, on a volume per volume basis. The preferred ratio of the pre-mixed oil containing phase to water is 4:1, respectively. Nanoparticles can be produced with reciprocating syringe instrumentation, continuous flow instrumentation, or high speed mixing equipment. Particles created at this 4:1 ratio range in diameters from 30 to 500 nanometers. These water miscible particles can then be filtered through either a 0.2 or 0.45 micron filter. Larger micellar particles can be created by simply increasing the water content. decreasing the oil-stabilizer-initiator content.
or changing the shear in forming the particles. We have coined the name "micellar nanoparticles" for particles with mean diameters less than 1000 nanometers (one micron).
'Trade Mark TABLE 1: Oils Utilized in Preparation of Micellar Nanoparticles .
Almond oil, sweet Apricot seed oil Borage oil Canola oil Coconut oil Corn oil Cotton seed oil Fish oil Jojoba bean oil Lard oil Linseed oil, boiled Macadamia nut oil Mineral oil Olive oil Peanut oil Safflower oil Sesame oil Soybean oil Squalane Sunflower seed oil Tricaprylin (1, ?, 3 trioctanoyl glycerol) Wheat germ oil -TABLE 2: Stabilizers/Surfactants Utilized in Preparation of Micellar Nanoparticles.
Tween#60 Tween#80 Nonvlphenol Polyethylene Glycol Ethers (alkylphenol-hydroxypolyoxyethylene) 1. Poly(oxy-1. 2-ethanedivl), alpha-(4-nonylphenol)-omega-hv_ droy-, branched (i.e. TergitoI~NP-40 Surfactant) Formula: C95H 1 g~040 MW (average) = 1980 1 ~ '_'. Nonylphenol Polyethylene Glycol Ether mixtures (i.e. TergitoI~NP-70 (70%AQ) Surfactant]
Formula and MV: not applicable (mixture) ?0 TABLE 3: Initiators Utilized in Preparation of Micellar Nanoparticles.
Ethanol Methanol 3~Trade Mark WO 96/23409 PCTlUS96/01410 _g_ TABLE 4: Flavored Initiators (flavored extracts*) Utilized in Preparation o f Micellar Nanoparticles.
Pure Anise extract (73% Ethanol) Imitation Banana extract (40% Ethanol) Imitation Chem~ extract (24% Ethanol) Chocolate extract (23% Ethanol) Pure Lemon extract (84% Ethanol) Pure Orange extract (80% Ethanol) Pure Peppermint extract (89% Ethanol) Imitation Pineapple extract (42% Ethanol) Imitation Rum extract (35% Ethanol) Imitation Strawberry extract (30% Ethanol) Pure Vanilla extract (35% Ethanol) * Extracts utilized are food grade materials (McCormick). Materials from other sources could be substituted.
TABLE 5: Volatile Oils or Fragrances Utilized in Preparation of Micellar Nanoparticles.
Balm oil Bay oil Bergamot oil Cedarwood oil Cherry oil Cinnamon oil 4~
Clove oil Origanum oil Peppermint oil WO 96/23409 PCTlUS96101410 -S-TABLE 6: Food Colors Utilized in Preparation of Micellar Nanoparticles.
Green Yellow Red Blue * Food colors utilized are food grade materials (McCormick). Materials from other sources could be substituted.
TABLE 7: List of Diluents Utilized in Preparation of Micellar Nanoparticles.
Water for injection Phosphate buffered saline The following Examples will more clearly illustrate the invention and its usefulness.
ExamQle 1- Production of Uncharged Micellar Nanooarticles Table 8 contains the materials used to produce micellar nanoparticles where water is the diluent. Sizing parameters using a Coulter L 130 Laser sizing apparatus are shown in Table 9.
TABLE 8: Preparation of Micellar nanoparticles utilizing water as the diluent.
Chemical Component Amount Soybean oil (Oil) '_'~ mL
Polysorbate 80 (Tween~80) (Stabilizer) 3 mL
Ethanol (Initiator) s mL
The above Oil-Stabilizer-Initiator components are mixed for 60 seconds. One mL
of water is injected into four mL of the mixture using reciprocating syringe instrumentation. This instrumentation has two ~mL syringes connected together through a stainless steel Leurlok connector with a 1/18,000 inch orifice. The solutions are driven between the syringes.
through the connector, for about 100 seconds. The resulting particles were dried on EM
grids, stained with uranyl acetate. and electron micrograph studies performed.
Figure 1 a shows an efectromicrograph of this preparation at a 60.OOOX magnification while Figure 1 b shows the same preparation at a 1 ~O,OOOX magnification. A brief description of the method of production of the micellar nanoparticles follows each table.
Table 9 - Sizing of Micellar Nanoparticles using water as a Diluent Mean Diameter Range Preparation ~nanometersl_ (nanomete~l_ Micellar nanoparticles 312 193-4s~
(SBOIT'v~0/E/WFI) One problem with using the LS 130 sizing device is that it cannot accurately size particles which are less than 200 nanometers in diameter. Using Figures 1 a and 1 b, it is determined that most of the particles are between 70 and 90 nanometers in diameter. with only ~% of particles be greater than 90 nanometers in diameter. Particles in the range of 20-30 nanometers are visible in the higher magnification shown in Figure lb.
3ETrade Mark Example 2 - Incorporation of Estradiot into Micellar Nanoparticles Tables 10 and 12 contain the materials utilized to produce two lots of uncharged a micellar nanoparticles into which estradiol has been incorporated at two different concentrations. Both preparations are made using water as the diluent. The higher estradiol concentration materials were used in the rhesus monkey studies described in Example 3 below . Either ~0 or 100 mg of estradiol is solublized in the initiator (ethanol component) of the preparation prior to formation of the micellar nanoparticles. This is necessar~~ since estradiol precipitates in the presence of water. In fact, the small amount of water in the reagent grade ethanol appears to be sufficient to precipitate the estradiol since the micellar particles formed using the materials and procedures described herein appear to have crystals of estradiol contained therein. However, these crystals appear to have a sheet-like form rather than the needle-like form standardly found in water precipitation.
TABLE 10: Preparation of Micellar Nanoparticles Containing Estradiol Soybean oil (Oil) 25mL
Polysorbate 80 (Tween 80) (Stabilizer) 3mL
Ethanol (Initiator) SmL
Estradiol SOmg The micellar nanoparticles were made using procedures substantially identical to that described in Example 1, except the estradiol was dissolved (or suspended) in the ethanol initiator prior to the mixing of the initiator with the other components. The oil-stabilizer-initiator/estradiol components are hand mixed or can be mixed for 60 seconds using a vortex mixer. One mL of water is injected into four mL of the resulting mixture using reciprocating syringe instrumentation such as is described in Example 1.
w TABLE 11 - Sizing data on Estradiol containing Micellar Nanoparticles (50 mg) Mean Diameter Range Preparation (nanometers) (nanometers) Micellar nanoparticles 289 174-459 (SBO/Tw#80/Etoh-estradiol/WFI) Sizing data on these preparations, measured using a Coulter LS130 Laser sizing apparatus. is shown in Tables 1 l and 13, respectively, for the two preparations.
The LS130 sizing device cannot size particles accurately less than 200 nanometers in diameter. These materials were also dried on EM grids, stained with uranyl acetate and electron micrograph studies performed. Electron micrographs reveal that most of the particles are less than 200 nanometers. Particles in the range of 20-30 nanometers are visible.
Crystallized estradiol is readily visible in the larger micelles. No free drug crystals are noted in any fields suggesting complete incorporation of drug into micelles.
TABLE 12: Preparation of Micellar Nanoparticles Containing Estradiol Chemical Component Amount Soybean oil (Oil) ?~mL
Polysorbate 80 (Tween~90) (Stabilizer) 3mL
Ethanol (Initiator) ~mL
Estradiol 100mg #Trade Mark ' -13-TABLE 13 -Sizing data on Estradiol containing Micellar Nanoparticles (100 mg) Mean Diameter Range Preparation ~nanometersl ~.anometersl Micellar nanoparticles 217 1 ~ 1-291 (SBO/Tw~O/Etoh-estradiol/WFI) Example 3 - Rhesus Monkey Testing of Estradiol Containing Preparations The 100 mg estradiol preparation of Example two was tested against a standard ethanol preparation of estradiol to show efficacy. One milligram of estradioh in either ethanol (Table 14) or micellar nanoparticles (Table 15), was applied to the skin of groups of four ovariectomized rhesus monkeys. Serial blood samples were drawn and serum estradiol levels were determined over the next 32 days. The serum estradiol data is graphically depicted in Figure ?. No additional drug was applied to skin of any animal.
Animals were observed for the next 60 davs to determine whether the time of occurrence.
duration and severity of vaginal bleeding (Table 16).
#Trade Mark WO 96/23409 PCT/tJS96/01410 TABLE 14 -Serum Estradiol Levels in Ovariectomized Female Monkeys Following a Single Topical Application of Micellar Nanoparticles Equivalent to 1 mg Estradiol Monkey Number k.
Sample Serum stradiol Group Mean E
Time #19567 #21792 #22366 #22405 S.E.
(pg/ml) (pg/ml)(pg/ml)(pg/ml) 0 hour 0.0b 0.0b 0.0b 0.0b 0.0 0.0h 0.5 hour 22.2 49.8 36.9 77.5 46.6 11.7 1 hour 37.4 60.9 65.6 108.6 68.1 14.8 2 hours 61.5 80.5 87.3 191.3 105.2 29.2 4 hours 77.2 132.1 120.6 120.4 112.6 12.1 6 hours 89.0 166.3 119.0 158.3 133.2 18.0 8 hours 87.5 157.3 116.1 148.1 127.3 15.9 12 hours 83.0 160.5 100.6 140.3 121.1 17.8 day 1 90.7 178.0 105.7 132.6 126.8 19.2 day 2 95.5 152.8 90.6 83.5 105.6 15.9 day 3 81.9 122.6 51.1 47.2 75.7 17.5 day 4 91.5 83.9 58.7 50.3 71.1 9.9 day ~ 41.6 74.7 3 5.1 40.0 47.9 9.1 day 6 45.2 63.7 25.6 40.9 43.9 7.8 day 7 18.3 25.9 21.9 27.0 23.3 2.0 daV 12 0.0b 0.0b 0.0b 0.0b 0.0 0.06 daV 17 0.0b 0.0b 0.0b 0.06 0.0 0.0b daV 22 0.0b 0.0b 0.0b 0.0b 0.0 0.06 daV 27 0.0b 0.0b 0.0b 0.0b 0.0 0.0b day 32 0.0b 0.0b 0.0b 0.0b 0.0 -~' 0.0b a CDB 3988 = 2.4 mg estradiol/ml of Tween/Oil. The dosing volume was 0.42 ml.
b 0 = Not Detectable. The limit of detection (ED9o) for the assay was 13.3 ~ 2.4 pg/ml (mean ~ S.E., n = 4) TABLE 15 - Serum Estradiol Levels in Ovariectomized Female Monkeys Following a Single Topical Application of 1 mg Ethanol Containing Estradiol$
Monkev Number Sample Serum Group Mean Estradiol Time #G-558 #G-603 #E-920 #E-924 S.E.
(pg/ml~(pg/ml) (pg/ml)(pg/ml) 0 hour 0.0b 0.06 0.0b 0.0b 0.0 0.06 0.5 hour 17.7 97.1 44.8 19.5 44.8 18.5 1 hour 53.2 44.1 88.3 99.9 71.4 13.5 2 hours 144.3 89.4 138.5 155.1 131.8 14.6 4 hours 143.7 202.3 165.1 193.6 176.2 13.4 6 hours 155.8 257.8 173.1 203.7 197.6 22.4 I
8 hours 114.2 266.1 130.7 130.0 160.3 35.5 12 hours 80.8 219.5 86.4 115.9 125.7 32.2 day 1 92.4 145.2 56.9 109.4 101.0 18.4 day 2 74.1 124.2 55.3 107.2 90.2 15.6 day 3 65.0 67.4 51.9 89.2 68.4 7.7 day 4 70.5 79.6 57.8 90.0 74.5 6.8 day 5 53.6 53.2 51.6 47.3 51.4 1.4 day 6 60.1 59.0 59.4 53.0 57.9 1.6 day 7 48.7 40.6 50.3 36.6 44.1 3.3 day 12 28.5 24.2 53.3 O.Ob 26.4 10.9b day 17 O.Ob O.Ob 28.9 O.Ob 7.2 7.2b day 22 O.Ob O.Ob 13.8 0.06 3.5 3.5 I, day 27 O.Ob 0.0b 0.0b 0.06 0.0 0.0b I'' day 32 0.06 0.0b 0.0b 0.0b 0.0 0 .0b a CDB 100 = 2.4 mg estradiol/ml of absolute ethanol. The dosing volume was 0.42 ml.
b 0 = Not Detectable. The limit of detection (ED9o) for the assay was 13.3 ~ 2.4 pg/ml (mean ~ S.E., n = 4) The data in Tables 14 and 1 ~ and Figure 2 show that therapeutic serum levels of estrogen are present in the blood stream of ovariectomized animals in both groups in one t hour after a single application. Mean estradiol levels greater than 40 picograms/ml are maintained for 7 days with the ethanol preparation and for 6 days with the nanoparticle r preparation. When estrogen levels remain low (see Figure 2 and Table 16), vaginal bleeding occurs in both groups. Also of particular interest is the shape of the curves in Figure 2. The ethanol-estradiol preparation yields a "shark tooth" curve showing a high initial action and a sharp fall-off while the micellar nanoparticle preparation yields more of a "mesa" effect with a nearly flat level for several hours. This "mesa" effect is often preferred since some of the problems associated with peaking can be minimized.
RHESUS MONKEYS FOLLOWING A SINGLE TOPICAL APPLICATION OF
ESTRADIOL IN ALCOHOL OR
MICELLAR NANOPARTICLES
WITHDRAWAL EDING
BLE
CDB No. ESTRADIOL ESTER DAYS INTENSITYa ' LATENCY DURATION
100 Estradiol in alcoholic19.5 0.3 4.3 0.9 1.6 0.2 solution 3988 Estradiol formulationb16.5 0.5~7.3 1.5 1.6 0.1 aMean intensity of bleeding (1=scant, moderate, 3=heavy) over bleeding period bNovavax MN Suspension 11294-2 Significantly different (p<0.01) from estradiol in alcohol solution based on a one-way analysis of variance followed by a Student Neuman-Keuls multiple range test Therefore, this Example demonstrates in a non-human primate that the micellar nanoparticles of the invention can be utilized to deliver estradiol through intact skin with maintenance of therapeutic serum estradiol levels for 6 days after a single application. This technology may have numerous therapeutic applications in medicine.
., 5 The estadiol preparation is also stable at a variety of temperatures. Table 17 shows thermal stability data for the micellar nanoparticle preparation of the Example 2 at -20°C, 25°C, and 65°C. As is clear, while the micellar nanoparticles are unstable at high temperatures, they are stable at room temperature and low temperatures.
TABLE 17: Thermal Stability of Micellar Nanoparticles Preparation Mean Range Diameter (nanometers) (nanometers) Micellar nanoparticles 361 168-599 (SBO/Tw80/Etoh-estradiol/WFI) Storage at 25°C
Micellar nanoparticles 312 179-510 (SBO/Tw80/Etoh-estradiol/WFI) Storage at -20°C
Micellar nanoparticles Unstable (SBO/Tw80/Etoh-estradiol/WFI) Storage at 65°C
In addition, the micellar nanoparticles of the invention can be diluted with aqueous solutions without stability loss. This allows the possibility of using high concentration products which can be diluted for use as necessary.
Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation. many equivalents to the specific embodiments of the invention described herein. Such equivalents are intended to be encompassed by the following claims.
a What is claimed is:
Claims (24)
1. A micellar nanoparticle having a diameter of between about 25 and 1000 nm, said micellar nanoparticle comprising a lipophilic phase which includes an oil, a stabilizer and an alcohol-based initiator, hydrated with a suitable aqueous-based solution wherein said stabilizer is selected from the group consisting of Tween*
60, Tween* 80, nonylphenol polyethylene glycol ethers, and mixtures thereof.
60, Tween* 80, nonylphenol polyethylene glycol ethers, and mixtures thereof.
2. The micellar nanoparticle of claim 1 wherein said initiator is selected from the group consisting of alcoholic-based materials containing methanol, ethanol and mixtures thereof.
3. The micellar nanoparticle of claim 2 wherein said initiator is selected from the group consisting of alcoholic-based materials containing 50%
or higher ethanol, methanol, and mixtures thereof.
or higher ethanol, methanol, and mixtures thereof.
4. The micellar nanoparticle of any one of claims 1 to 3 wherein said oil is selected from the group consisting of vegetable oils, nut oils, fish oils, lard oil, mineral oils, squalane tricaprylin, and mixtures thereof.
5. The micellar nanoparticle of any one of claims 1 to 4 wherein said aqueous solution comprises a physiologically compatible solution.
6. The micellar nanoparticle of claim 5 wherein said aqueous solution is selected from the group consisting of water, and phosphate buffered saline.
7. The micellar nanoparticle of any one of claims 1 to 6 wherein said aqueous solution has an active material dissolved or suspended therein.
8. The micellar nanoparticle of any one of claims 1 to 7 wherein said oil has an active material dissolved or suspended therein.
9. The micellar nanoparticle of any one of claims 1 to 9 wherein said initiator has an active material dissolved or suspended therein.
10. The micellar nanoparticle of claim 9 wherein said active material comprises estradiol.
11. The micellar nanoparticle of any one of claims 1 to 10 wherein said micellar nanoparticle is dispersible in aqueous solution.
12. The micellar nanoparticle of any one of claims 1 to 11 wherein the diameter of said micellar nanoparticle allows passage through a 0.2 mm filter.
13. A method of making micellar nanoparticles comprising the steps of:
blending an excess of an oil, together with a stabilizer and an initiator to form a lipophilic phase, said stabilizer being selected from the group consisting of Tween* 60, Tween* 80, Nonylphenol Polyethylene Glycol Ethers, and mixtures thereof;
preparing a dilulent solution having an aqueous solution base; and blending an excess of said lipophilic phase with said dilulent to form said micellar nanoparticles.
blending an excess of an oil, together with a stabilizer and an initiator to form a lipophilic phase, said stabilizer being selected from the group consisting of Tween* 60, Tween* 80, Nonylphenol Polyethylene Glycol Ethers, and mixtures thereof;
preparing a dilulent solution having an aqueous solution base; and blending an excess of said lipophilic phase with said dilulent to form said micellar nanoparticles.
14. The method of claim 13 wherein said initiator is selected from the group consisting of alcoholic-based materials containing methanol, ethanol and mixtures thereof.
15. The method of claim 14 wherein said initiator is selected from the group consisting of alcoholic-based materials containing 50% or higher ethanol, methanol, and mixtures thereof.
16. The method of any one of claims 13 to 15 wherein said oil is selected from the group consisting of vegetable oils, nut oils, fish oils, lard oil, mineral oils, squalane, tricaprylin, and mixtures thereof.
17. The method of any one of claims 13 to 16 wherein said aqueous solution comprises a physiologically compatible solution.
18. The method of claim 17 wherein said aqueous solution is selected from the group consisting of water, and phosphate buffered saline.
19. The method of any one of claims 13 to 18 wherein said aqueous solution has an active material dissolved or suspended therein.
20. The method of any one of claims 13 to 19 wherein said oil has an active material dissolved or suspended therein.
21. The method of any one of claims 13 to 20 wherein said initiator has an active material dissolved or suspended therein.
22. The method of claim 21 wherein said active material comprises estradiol.
23. The method of any one of claims 13 to 22 wherein said blending of said lipophilic phase and said dilulent is achieved using a relative velocity of about 50 m/s through a 1/18,000 inch orifice.
24. The method of any one of claims 13 to 23 wherein the ratio of said lipophilic phase to said aqueous phase is about 4:1.
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| US08/380,942 US5629021A (en) | 1995-01-31 | 1995-01-31 | Micellar nanoparticles |
| US08/380,942 | 1995-01-31 | ||
| PCT/US1996/001410 WO1996023409A1 (en) | 1995-01-31 | 1996-01-29 | Micellar nanoparticles |
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|---|---|---|---|---|
| US6387373B1 (en) * | 1993-01-15 | 2002-05-14 | Novavax, Inc. | Vaccines containing paucilsmellar lipid vesicles as immunological adjuvants |
| US6542183B1 (en) | 1995-06-28 | 2003-04-01 | Lynx Systems Developers, Inc. | Event recording apparatus |
| US5766629A (en) | 1995-08-25 | 1998-06-16 | Sangstat Medical Corporation | Oral cyclosporin formulations |
| US6958148B1 (en) * | 1998-01-20 | 2005-10-25 | Pericor Science, Inc. | Linkage of agents to body tissue using microparticles and transglutaminase |
| KR100342835B1 (en) * | 1998-07-31 | 2002-07-02 | 박호군 | Lipid emulsion as a gene or drug carrier, formulation, and method for preparating thereof |
| US6284234B1 (en) * | 1998-08-04 | 2001-09-04 | Johnson & Johnson Consumer Companies, Inc. | Topical delivery systems for active agents |
| US20040258763A1 (en) * | 1999-02-03 | 2004-12-23 | Bell Steve J.D. | Methods of manufacture and use of calcium phosphate particles containing allergens |
| US20020054914A1 (en) * | 1999-02-03 | 2002-05-09 | Tulin Morcol | Compositions and methods for therapuetic agents complexed with calcium phosphate and encased by casein |
| ATE276199T1 (en) * | 1999-02-03 | 2004-10-15 | Biosante Pharmaceuticals Inc | METHODS FOR PRODUCING THERAPEUTIC CALCIUM PHOSPHATE PARTICLES |
| US7374779B2 (en) * | 1999-02-26 | 2008-05-20 | Lipocine, Inc. | Pharmaceutical formulations and systems for improved absorption and multistage release of active agents |
| US6248363B1 (en) * | 1999-11-23 | 2001-06-19 | Lipocine, Inc. | Solid carriers for improved delivery of active ingredients in pharmaceutical compositions |
| US7655252B2 (en) | 1999-04-28 | 2010-02-02 | The Regents Of The University Of Michigan | Antimicrobial nanoemulsion compositions and methods |
| US8232320B2 (en) | 1999-04-28 | 2012-07-31 | 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 |
| US20030236236A1 (en) * | 1999-06-30 | 2003-12-25 | Feng-Jing Chen | Pharmaceutical compositions and dosage forms for administration of hydrophobic drugs |
| CA2400172C (en) | 2000-02-28 | 2010-04-20 | Genesegues, Inc. | Nanocapsule encapsulation system and method |
| US20070225379A1 (en) * | 2001-08-03 | 2007-09-27 | Carrara Dario Norberto R | Transdermal delivery of systemically active central nervous system drugs |
| US8980290B2 (en) | 2000-08-03 | 2015-03-17 | Antares Pharma Ipl Ag | Transdermal compositions for anticholinergic agents |
| US7198801B2 (en) * | 2000-08-03 | 2007-04-03 | Antares Pharma Ipl Ag | Formulations for transdermal or transmucosal application |
| US20040198706A1 (en) * | 2003-03-11 | 2004-10-07 | Carrara Dario Norberto R. | Methods and formulations for transdermal or transmucosal application of active agents |
| US7214381B2 (en) | 2000-08-03 | 2007-05-08 | Antares Pharma Ipl Ag | Composition for transdermal and/or transmucosal administration of active compounds that ensures adequate therapeutic levels |
| US6423707B1 (en) | 2000-08-28 | 2002-07-23 | California Pacific Medical Center | Nitroimidazole ester analogues and therapeutic applications |
| US6503894B1 (en) | 2000-08-30 | 2003-01-07 | Unimed Pharmaceuticals, Inc. | Pharmaceutical composition and method for treating hypogonadism |
| US20040002482A1 (en) * | 2000-08-30 | 2004-01-01 | Dudley Robert E. | Androgen pharmaceutical composition and method for treating depression |
| US20040092494A9 (en) * | 2000-08-30 | 2004-05-13 | Dudley Robert E. | Method of increasing testosterone and related steroid concentrations in women |
| US6537591B2 (en) | 2001-03-02 | 2003-03-25 | Aquarium Pharmaceuticals, Inc. | Aquatic animal treatment method and composition containing Pimenta extract |
| US20030185892A1 (en) * | 2001-08-17 | 2003-10-02 | Bell Steve J. D. | Intraocular delivery compositions and methods |
| NZ531492A (en) * | 2001-08-28 | 2007-11-30 | Eden Research Plc | Treatment and prevention of fungal, bacterial, mycoplasma and phytoplasma infections in plants comprising terpene compounds in true solution |
| US6878693B2 (en) * | 2001-09-28 | 2005-04-12 | Solubest Ltd. | Hydrophilic complexes of lipophilic materials and an apparatus and method for their production |
| US20050227911A1 (en) * | 2001-09-28 | 2005-10-13 | Solubest Ltd. | Hydrophilic dispersions of nanoparticles of inclusion complexes of macromolecules |
| US20050233003A1 (en) * | 2001-09-28 | 2005-10-20 | Solubest Ltd. | Hydrophilic dispersions of nanoparticles of inclusion complexes of salicylic acid |
| US8765167B2 (en) | 2001-10-12 | 2014-07-01 | Monosol Rx, Llc | Uniform films for rapid-dissolve dosage form incorporating anti-tacking compositions |
| US8663687B2 (en) * | 2001-10-12 | 2014-03-04 | Monosol Rx, Llc | Film compositions for delivery of actives |
| US10285910B2 (en) | 2001-10-12 | 2019-05-14 | Aquestive Therapeutics, Inc. | Sublingual and buccal film compositions |
| US20190328679A1 (en) | 2001-10-12 | 2019-10-31 | Aquestive Therapeutics, Inc. | Uniform films for rapid-dissolve dosage form incorporating anti-tacking compositions |
| US11207805B2 (en) | 2001-10-12 | 2021-12-28 | Aquestive Therapeutics, Inc. | Process for manufacturing a resulting pharmaceutical film |
| US7357891B2 (en) | 2001-10-12 | 2008-04-15 | Monosol Rx, Llc | Process for making an ingestible film |
| US20110033542A1 (en) | 2009-08-07 | 2011-02-10 | Monosol Rx, Llc | Sublingual and buccal film compositions |
| US8603514B2 (en) | 2002-04-11 | 2013-12-10 | Monosol Rx, Llc | Uniform films for rapid dissolve dosage form incorporating taste-masking compositions |
| US20070281003A1 (en) | 2001-10-12 | 2007-12-06 | Fuisz Richard C | Polymer-Based Films and Drug Delivery Systems Made Therefrom |
| US8900498B2 (en) | 2001-10-12 | 2014-12-02 | Monosol Rx, Llc | Process for manufacturing a resulting multi-layer pharmaceutical film |
| US8900497B2 (en) | 2001-10-12 | 2014-12-02 | Monosol Rx, Llc | Process for making a film having a substantially uniform distribution of components |
| US7700851B2 (en) * | 2001-11-13 | 2010-04-20 | U.S. Smokeless Tobacco Company | Tobacco nicotine demethylase genomic clone and uses thereof |
| EP1455768A1 (en) * | 2001-12-07 | 2004-09-15 | Eden Research Plc | Respiratory infection prevention and treatment with terpene-containing compositions |
| US7763663B2 (en) * | 2001-12-19 | 2010-07-27 | University Of Massachusetts | Polysaccharide-containing block copolymer particles and uses thereof |
| WO2003070181A2 (en) * | 2002-02-19 | 2003-08-28 | Eden Research Plc | Compositions and methods for preservation of food |
| WO2003070286A1 (en) * | 2002-02-19 | 2003-08-28 | Eden Research Plc | Improvement of indoor air quality and antiseptic composition for use therein |
| US20040038303A1 (en) * | 2002-04-08 | 2004-02-26 | Unger Gretchen M. | Biologic modulations with nanoparticles |
| 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 |
| ATE534373T1 (en) * | 2003-10-10 | 2011-12-15 | Antares Pharma Ipl Ag | TRANSDERMAL PHARMACEUTICAL FORMULATION TO MINIMIZE RESIDUE ON THE SKIN |
| US20050084456A1 (en) * | 2003-10-21 | 2005-04-21 | Liping Tang | Functionalized particles |
| US20050084513A1 (en) * | 2003-10-21 | 2005-04-21 | Board Of Regents | Nanocoating for improving biocompatibility of medical implants |
| WO2005070213A2 (en) | 2004-01-23 | 2005-08-04 | Eden Research Plc | Methods of killing nematodes comprising the application of a terpene component |
| CN1993113B (en) * | 2004-02-13 | 2010-12-08 | Nod药物公司 | Particles comprising a core of calcium phosphate nanoparticles, a biomolecule and a bile acid, methods of manufacturing, therapeutic use thereof |
| US7425340B2 (en) * | 2004-05-07 | 2008-09-16 | Antares Pharma Ipl Ag | Permeation enhancing compositions for anticholinergic agents |
| ES2461178T3 (en) | 2004-05-20 | 2014-05-19 | Eden Research Plc | Hollow glucan or cell wall particle that encapsulates a terpene component |
| US9801982B2 (en) | 2004-09-28 | 2017-10-31 | Atrium Medical Corporation | Implantable barrier device |
| US9012506B2 (en) | 2004-09-28 | 2015-04-21 | Atrium Medical Corporation | Cross-linked fatty acid-based biomaterials |
| US9000040B2 (en) | 2004-09-28 | 2015-04-07 | Atrium Medical Corporation | Cross-linked fatty acid-based biomaterials |
| US8263102B2 (en) | 2004-09-28 | 2012-09-11 | Atrium Medical Corporation | Drug delivery coating for use with a stent |
| US20060067976A1 (en) | 2004-09-28 | 2006-03-30 | Atrium Medical Corporation | Formation of barrier layer |
| CA2586035A1 (en) | 2004-11-01 | 2006-05-11 | Biosante Pharmaceuticals, Inc. | Therapeutic calcium phosphate particles in use for aesthetic or cosmetic medicine, and methods of manufacture and use |
| US20070088012A1 (en) * | 2005-04-08 | 2007-04-19 | Woun Seo | Method of treating or preventing type-2 diabetes |
| US7371738B2 (en) * | 2005-04-15 | 2008-05-13 | University Of South Florida | Method of transdermal drug delivery using hyaluronic acid nanoparticles |
| WO2007124250A2 (en) | 2006-04-21 | 2007-11-01 | Antares Pharma Ipl Ag | Methods of treating hot flashes with formulations for transdermal or transmucosal application |
| WO2006125642A1 (en) | 2005-05-27 | 2006-11-30 | Antares Pharma Ipl Ag | Methods and apparatus for transdermal or transmucosal application of testosterone |
| US20080274195A1 (en) | 2005-07-18 | 2008-11-06 | University Of Massachusetts Lowell | Compositions and Methods for Making and Using Nanoemulsions |
| US9278161B2 (en) | 2005-09-28 | 2016-03-08 | Atrium Medical Corporation | Tissue-separating fatty acid adhesion barrier |
| US9427423B2 (en) | 2009-03-10 | 2016-08-30 | Atrium Medical Corporation | Fatty-acid based particles |
| AU2006299833B2 (en) | 2005-10-12 | 2012-04-12 | Besins Healthcare Luxembourg Sarl | Improved testosterone gel and method of use |
| BRPI0617486A2 (en) | 2005-10-18 | 2011-07-26 | Novavax Inc | vaccine comprising functional influenza virus (vlp) -like particles and use of an influenza vlp |
| US10667512B2 (en) | 2005-11-30 | 2020-06-02 | Eden Research Plc | Terpene-containing compositions and methods of making and using them |
| AP2901A (en) | 2005-11-30 | 2014-05-31 | Eden Research Plc | Compositions and methods comprising terpenes or terpene mixtures selected form thymol, eugenol, geraniol, citral, and L-carvone |
| US9486408B2 (en) | 2005-12-01 | 2016-11-08 | University Of Massachusetts Lowell | Botulinum nanoemulsions |
| PT3144013T (en) | 2005-12-01 | 2019-02-07 | Univ Massachusetts Lowell | Botulinum nanoemulsions |
| US11311477B2 (en) | 2006-03-07 | 2022-04-26 | Sgn Nanopharma Inc. | Ophthalmic preparations |
| US10137083B2 (en) * | 2006-03-07 | 2018-11-27 | SGN Nanopharma Inc | Ophthalmic preparations |
| WO2007103294A2 (en) * | 2006-03-07 | 2007-09-13 | Novavax, Inc. | Nanoemulsions of poorly soluble pharmaceutical active ingredients and methods of making the same |
| EP2010149A1 (en) * | 2006-04-07 | 2009-01-07 | Novavax, Inc. | Nanostructured compositions having antibacterial, anti-fungal, anti-yeast, and/or anti-viral properties |
| US9040250B2 (en) * | 2006-09-07 | 2015-05-26 | The United States Of America, As Represented By The Secretary Of The Navy | Binding interaction of proanthocyanidins with bacteria and bacterial components |
| WO2008039864A1 (en) * | 2006-09-26 | 2008-04-03 | Novavax, Inc. | Emulsion formulations for transdermal delivery of poorly water soluble active agents |
| EP2083875B1 (en) | 2006-11-06 | 2013-03-27 | Atrium Medical Corporation | Coated surgical mesh |
| US9492596B2 (en) | 2006-11-06 | 2016-11-15 | Atrium Medical Corporation | Barrier layer with underlying medical device and one or more reinforcing support structures |
| CA2671133C (en) * | 2006-12-01 | 2015-11-24 | Anterios, Inc. | Peptide nanoparticles and uses therefor |
| US20100150994A1 (en) | 2006-12-01 | 2010-06-17 | Anterios, Inc. | Amphiphilic entity nanoparticles |
| WO2008067991A2 (en) * | 2006-12-08 | 2008-06-12 | Antares Pharma Ipl Ag | Skin-friendly drug complexes for transdermal administration |
| US20080207756A1 (en) * | 2007-02-27 | 2008-08-28 | Atrium Medical Corporation | Bio-absorbable oil suspension |
| WO2008137747A1 (en) | 2007-05-02 | 2008-11-13 | The Regents Of The University Of Michigan | Nanoemulsion therapeutic compositions and methods of using the same |
| CN101765423B (en) | 2007-05-31 | 2014-08-06 | 安特里奥公司 | Nucleic acid nanoparticles and uses therefor |
| PT2540312E (en) | 2007-07-19 | 2015-08-31 | Novavax Inc | Avian influenza chimeric vlps |
| WO2009158687A1 (en) * | 2008-06-26 | 2009-12-30 | Anterios, Inc. | Dermal delivery |
| WO2010057197A1 (en) | 2008-11-17 | 2010-05-20 | The Regents Of The University Of Michigan | Cancer vaccine compositions and methods of using the same |
| HUE051666T2 (en) | 2008-12-09 | 2021-03-29 | Novavax Inc | Modified rsv f proteins and methods of their use |
| US11304960B2 (en) | 2009-01-08 | 2022-04-19 | Chandrashekar Giliyar | Steroidal compositions |
| WO2010116382A2 (en) | 2009-04-08 | 2010-10-14 | Cadila Healthcare Limited | Stable pharmaceutical compositions of diclofenac |
| US8470304B2 (en) | 2009-08-04 | 2013-06-25 | Avidas Pharmaceuticals Llc | Therapeutic vitamin D sun-protecting formulations and methods for their use |
| US20110038910A1 (en) | 2009-08-11 | 2011-02-17 | Atrium Medical Corporation | Anti-infective antimicrobial-containing biomaterials |
| US20110045050A1 (en) * | 2009-08-24 | 2011-02-24 | Atrium Medical Corporation | Nanoemulsion formulations for direct delivery |
| WO2012009707A2 (en) | 2010-07-16 | 2012-01-19 | Atrium Medical Corporation | Composition and methods for altering the rate of hydrolysis of cured oil-based materials |
| US10028972B2 (en) | 2010-10-21 | 2018-07-24 | Cadila Healthcare Limited | Pharmaceutical compositions of anti-acne agents |
| WO2012053009A2 (en) | 2010-10-21 | 2012-04-26 | Cadila Healthcare Limited | Pharmaceutical compositions comprising skin whitening agents |
| WO2012053012A2 (en) | 2010-10-21 | 2012-04-26 | Cadila Healthcare Limited | Topical pharmaceutical compositions comprising tfflocolchicoside |
| WO2012053007A1 (en) | 2010-10-21 | 2012-04-26 | Cadila Healthcare Limited | Topical pharmaceutical compositions containing nanodroplets for the treatment psoriasis |
| US20130295145A1 (en) | 2010-10-21 | 2013-11-07 | Cadila Healthcare Limited | Topical pharmaceutical compositions comprising etodolac |
| US9789123B2 (en) | 2010-10-21 | 2017-10-17 | Cadila Healthcare Limited | Topical pharmaceutical compositions containing nanodroplets for the treatment of psoriasis |
| US9149959B2 (en) | 2010-10-22 | 2015-10-06 | Monosol Rx, Llc | Manufacturing of small film strips |
| WO2012059936A1 (en) | 2010-11-03 | 2012-05-10 | Padma Venkitachalam Devarajan | Pharmaceutical compositions for colloidal drug delivery |
| MX354750B (en) | 2010-11-05 | 2018-03-20 | Novavax Inc | RABIES GLYCOPROTEIN VIRUS-LIKE PARTICLES (VLPs). |
| US20180153904A1 (en) | 2010-11-30 | 2018-06-07 | Lipocine Inc. | High-strength testosterone undecanoate compositions |
| US9358241B2 (en) | 2010-11-30 | 2016-06-07 | Lipocine Inc. | High-strength testosterone undecanoate compositions |
| US9034858B2 (en) | 2010-11-30 | 2015-05-19 | Lipocine Inc. | High-strength testosterone undecanoate compositions |
| US20120148675A1 (en) | 2010-12-10 | 2012-06-14 | Basawaraj Chickmath | Testosterone undecanoate compositions |
| EP3263121B1 (en) | 2010-12-21 | 2019-02-27 | Leidos, Inc. | Methods and compositions for wound treatment |
| US20120189677A1 (en) * | 2011-01-20 | 2012-07-26 | Stephen Tonge | Formulations |
| WO2012127496A1 (en) | 2011-03-01 | 2012-09-27 | Cadila Healthcare Limited | Stable pharmaceutical compositions of lornoxicam or salts thereof |
| CN104023717B (en) | 2011-06-29 | 2017-08-04 | 阿维达斯制药有限责任公司 | The topical formulation and its purposes of mediator are delivered including lipid microcapsules |
| WO2013014680A1 (en) | 2011-07-28 | 2013-01-31 | Cadila Healthcare Limited | Method for treatment of pain and inflammation |
| PT2782584T (en) | 2011-11-23 | 2021-09-02 | Therapeuticsmd Inc | Natural combination hormone replacement formulations and therapies |
| US9301920B2 (en) | 2012-06-18 | 2016-04-05 | Therapeuticsmd, Inc. | Natural combination hormone replacement formulations and therapies |
| US10322089B2 (en) * | 2012-03-14 | 2019-06-18 | The Board Of Trustees Of The Leland Stanford Junior University | Nanoparticles, nanoparticle delivery methods, and systems of delivery |
| US9867880B2 (en) | 2012-06-13 | 2018-01-16 | Atrium Medical Corporation | Cured oil-hydrogel biomaterial compositions for controlled drug delivery |
| US10806697B2 (en) | 2012-12-21 | 2020-10-20 | Therapeuticsmd, Inc. | Vaginal inserted estradiol pharmaceutical compositions and methods |
| US20130338122A1 (en) | 2012-06-18 | 2013-12-19 | Therapeuticsmd, Inc. | Transdermal hormone replacement therapies |
| US20150196640A1 (en) | 2012-06-18 | 2015-07-16 | Therapeuticsmd, Inc. | Progesterone formulations having a desirable pk profile |
| US10806740B2 (en) | 2012-06-18 | 2020-10-20 | Therapeuticsmd, Inc. | Natural combination hormone replacement formulations and therapies |
| GB201220940D0 (en) | 2012-11-21 | 2013-01-02 | Eden Research Plc | Method P |
| US9180091B2 (en) | 2012-12-21 | 2015-11-10 | Therapeuticsmd, Inc. | Soluble estradiol capsule for vaginal insertion |
| US10568891B2 (en) | 2012-12-21 | 2020-02-25 | Therapeuticsmd, Inc. | Vaginal inserted estradiol pharmaceutical compositions and methods |
| US11266661B2 (en) | 2012-12-21 | 2022-03-08 | Therapeuticsmd, Inc. | Vaginal inserted estradiol pharmaceutical compositions and methods |
| US10471072B2 (en) | 2012-12-21 | 2019-11-12 | Therapeuticsmd, Inc. | Vaginal inserted estradiol pharmaceutical compositions and methods |
| US11246875B2 (en) | 2012-12-21 | 2022-02-15 | Therapeuticsmd, Inc. | Vaginal inserted estradiol pharmaceutical compositions and methods |
| US10537581B2 (en) | 2012-12-21 | 2020-01-21 | Therapeuticsmd, Inc. | Vaginal inserted estradiol pharmaceutical compositions and methods |
| MX2016003419A (en) | 2013-09-19 | 2016-10-10 | Novavax Inc | Immunogenic middle east respiratory syndrome coronavirus (mers-cov) compositions and methods. |
| AR100562A1 (en) | 2014-05-22 | 2016-10-12 | Therapeuticsmd Inc | PHARMACEUTICAL COMPOSITION OF ESTRADIOL AND PROGESTERONE FOR HORMONAL REPLACEMENT THERAPY |
| WO2016033549A2 (en) | 2014-08-28 | 2016-03-03 | Lipocine Inc. | (17-ß)-3-OXOANDROST-4-EN-17-YL TRIDECANOATE COMPOSITIONS AND METHODS OF THEIR PREPARATION AND USE |
| US9498485B2 (en) | 2014-08-28 | 2016-11-22 | Lipocine Inc. | Bioavailable solid state (17-β)-hydroxy-4-androsten-3-one esters |
| SG10201902499VA (en) | 2014-09-03 | 2019-04-29 | Genesegues Inc | Therapeutic nanoparticles and related compositions, methods and systems |
| US10894024B2 (en) * | 2014-12-08 | 2021-01-19 | Synaptamine, Inc. | Anti-RDS compounds and method of manufacture and administration thereof to induce dopamine homeostatis |
| US10328087B2 (en) | 2015-07-23 | 2019-06-25 | Therapeuticsmd, Inc. | Formulations for solubilizing hormones |
| CN114699519A (en) | 2015-09-03 | 2022-07-05 | 诺瓦瓦克斯股份有限公司 | Vaccine compositions with improved stability and immunogenicity |
| WO2017173044A1 (en) | 2016-04-01 | 2017-10-05 | Therapeuticsmd Inc. | Steroid hormone compositions in medium chain oils |
| CA3020153A1 (en) | 2016-04-01 | 2017-10-05 | Therapeuticsmd, Inc. | Steroid hormone pharmaceutical composition |
| US11273131B2 (en) | 2016-05-05 | 2022-03-15 | Aquestive Therapeutics, Inc. | Pharmaceutical compositions with enhanced permeation |
| BR112018072539A2 (en) | 2016-05-05 | 2019-03-26 | Aquestive Therapeutics, Inc. | increased administration epinephrine compositions |
| BR112019010131A2 (en) | 2016-11-21 | 2019-10-08 | Eirion Therapeutics Inc | transdermal delivery of large agents |
| JP2020503269A (en) | 2016-11-28 | 2020-01-30 | リポカイン インコーポレーテッド | Oral testosterone undecanoate therapy |
| WO2019023196A1 (en) | 2017-07-24 | 2019-01-31 | Novavax, Inc. | Methods and compositions for treating respiratory disease |
| CN110881462A (en) * | 2019-11-25 | 2020-03-17 | 南京启佑生物科技有限公司 | Nano pesticide composite preparation based on phosphate and metal ions and preparation method thereof |
| US11633405B2 (en) | 2020-02-07 | 2023-04-25 | Therapeuticsmd, Inc. | Steroid hormone pharmaceutical formulations |
Family Cites Families (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4182330A (en) * | 1977-07-25 | 1980-01-08 | Alza Corporation | Means for administering amphipathic medicament |
| US4356167A (en) * | 1978-01-27 | 1982-10-26 | Sandoz, Inc. | Liposome drug delivery systems |
| US4235871A (en) * | 1978-02-24 | 1980-11-25 | Papahadjopoulos Demetrios P | Method of encapsulating biologically active materials in lipid vesicles |
| DE3161962D1 (en) * | 1980-07-01 | 1984-02-23 | Oreal | Process for obtaining stable dispersions in an aqueous phase of at least a water immiscible liquid phase, and corresponding dispersions |
| US4725442A (en) * | 1983-06-17 | 1988-02-16 | Haynes Duncan H | Microdroplets of water-insoluble drugs and injectable formulations containing same |
| US4744989A (en) * | 1984-02-08 | 1988-05-17 | E. R. Squibb & Sons, Inc. | Method of preparing liposomes and products produced thereby |
| US4610868A (en) * | 1984-03-20 | 1986-09-09 | The Liposome Company, Inc. | Lipid matrix carriers for use in drug delivery systems |
| US4911928A (en) * | 1987-03-13 | 1990-03-27 | Micro-Pak, Inc. | Paucilamellar lipid vesicles |
| US4824675A (en) * | 1987-07-13 | 1989-04-25 | Alza Corporation | Dispenser with movable matrix comprising a plurality of tiny pills |
| DE3919982A1 (en) * | 1989-06-19 | 1990-12-20 | Liedtke Pharmed Gmbh | ORAL LIPID MEDICINE FORM |
| CH677886A5 (en) * | 1989-06-26 | 1991-07-15 | Hans Georg Prof Dr Weder | |
| AU658608B2 (en) * | 1991-03-25 | 1995-04-27 | Astellas Pharma Europe B.V. | Topical preparation containing a suspension of solid lipid particles |
-
1995
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1996
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- 1996-01-29 CA CA002211262A patent/CA2211262C/en not_active Expired - Fee Related
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- 1996-01-29 JP JP52373596A patent/JP3974170B2/en not_active Expired - Fee Related
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- 1996-01-31 ZA ZA96738A patent/ZA96738B/en unknown
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| JPH10513185A (en) | 1998-12-15 |
| DE69627309D1 (en) | 2003-05-15 |
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