|Publication number||WO1991005548 A1|
|Publication date||2 May 1991|
|Filing date||20 Sep 1990|
|Priority date||10 Oct 1989|
|Publication number||PCT/1990/5345, PCT/US/1990/005345, PCT/US/1990/05345, PCT/US/90/005345, PCT/US/90/05345, PCT/US1990/005345, PCT/US1990/05345, PCT/US1990005345, PCT/US199005345, PCT/US90/005345, PCT/US90/05345, PCT/US90005345, PCT/US9005345, WO 1991/005548 A1, WO 1991005548 A1, WO 1991005548A1, WO 9105548 A1, WO 9105548A1, WO-A1-1991005548, WO-A1-9105548, WO1991/005548A1, WO1991005548 A1, WO1991005548A1, WO9105548 A1, WO9105548A1|
|Inventors||Kallidaikurichi N. Sivaramakrishnan, Matthew W. Gray|
|Export Citation||BiBTeX, EndNote, RefMan|
|Patent Citations (3), Referenced by (212), Classifications (10), Legal Events (3)|
|External Links: Patentscope, Espacenet|
Sustained Release Composition for Macromolecular Proteins
This invention relates generally to sustained release compositions and particularly to a sustained release composition for the prolonged release of macromolecular proteins.
Background of the Invention The delivery of macromolecular proteins to animals is complicated by the need for delivery devices which will maintain the bioactivity of the protein prior to, during, and after administration to the animal. Merely exposing macromolecular proteins to aqueous environments causes the large proteins to form precipitates and aggregates which destroy the bioactivity and therefore the usefulness of the protein. Also, hydrolytic reactions resulting from "wetting" the proteins may produce bio-inactive products which reduce the .in vivo effectiveness of the protein.
Prior art methods for overcoming this problem have included encapsulating the macromolecular proteins in polymer-coated tablets which limit the ingress of water, pressing the macromolecular proteins into pellet compositions which control the release of the protein by limiting the surface area of the pellet and therefore the amount of water that has access to the protein, placing the macromolecular proteins in porous devices which limit the ability of water to enter the device and the ability of the protein to exit the device, and the like. Patents and other references describing these methods are voluminous and well known in the art.
Sustained release of drugs can be achieved by dispersing or dissolving them in a water-insoluble matrix. Suitable matrix materials for such diffusion-controlled delivery devices include polymers such as poly(ethylene-co-vinyl acetate) (EVA), ethyl cellulose, and polysiloxanes. The mechanism of release is the slow diffusion of the dissolved drug through interconnecting pores and channels formed in the matrix. See, Rhine et al. , J. Pharm. Sci. , Vol. 69(3), p. 265 (1980) and Goodhart et al. , J. Pharm. Sci., Vol. 63(11), p. 1748 (1974). The release characteristics of these devices are determined by the physical properties such as visco-elasticity, glass transition temperature, and porosity of the matrix.
Waxes have been used in solid dosage forms as lubricants, coatings, and inert fillers. Appropriate formulation of the drug in a wax matrix can give rise to sustained release of the drug. A sustained release dosage form of tripelennamine hydrochloride was developed using carnauba wax as the matrix; Dakkuri et al. , J. Pharm. Sci., Vol. 67(3), p. 354 (1978). The drug was dispersed along with a surfactant in molten carnauba wax containing stearyl alcohol. The congealed mass was granulated and compressed into cores. Hydrophobic surfactants such as glycerol monostearate did not affect the dissolution rate of the tablets. On the other hand, hydrophilic surfactants such as polyoxyethylene lauryl ether had a profound effect, releasing about 80% of the drug in about 4 hours. The data were interpreted in terms of facile wetting of the matrix, promoted by the surfactant, thus creating channels for the diffusion of the drug. Inclusion of polyvinylpyrrolidone in the wax matrix accomplished the same result.
Controlled delivery of indomethacin was achieved from carnauba wax matrix containing surfactants. A combination of erosion, leaching, and solubilization of the drug was used to explain the sustained release of sulfaethylthiadiazole from tablets made by compressing a spray-congealed wax matrix; Hamid et al. , J. Pharm. Sci. , Vol. 59 (4), p. 511 (1970). Sustained release of drugs could also be obtained through "slowly disintegrating tablets".as detailed in the U.S. Patent 4,695,467. Sustained elevation of plasma growth hormone level from a soybean oil/beeswax implant has been reported; Davis et al. , J. Dairy Sci. , Vol. 66, p. 1980-2 (1983).
Also, U.S. Patent No. 4,404,183 discloses a sustained release composition of solid medical material which has water-soluble surfactants such as polyethylene glycol as a component. U.S. Patent No. 4,404,183 discloses a controlled release composition which may contain surfactants which are soluble in water and in solvent and are stable at high temperatures. Neither of these patents, however, discloses the use of a water-insoluble surfactant. Schroeder et al. , J. Pharm. Sci. , Vol. 67, p. 350 (1978) discloses sustained release combinations of small organic drugs such as tripelennamine hydrochloride or tolazoline hydrochloride from wax matrixes. Similarly, Hamid et al. , J. Pharm. Sci. , Vol. 59 (4), p. 511 (1970) discloses sustained release compositions containing a small organic drug sulfaethylthiadiazole in wax matrixes. The compositions did not, however, contain surfactants nor provide release characteristics for macromolecular proteins.
Al-shora et al. , Int. J. Pharm. , Vol. 7, p. 77 (1980) discloses using wax matrixes for producing sustained release compositions containing small organic drugs such as tripelennamine hydrochloride and water-soluble channeling agents such as polyethylene glycol. The compositions did not, however, contain water-insoluble surfactants nor provide a sustained release composition for macromolecular proteins. Because of the problems associated with prior sustained release compositions and devices for macromolecular proteins, there exists a continuing need for new sustained release compositions which control the manner and timing of delivery while maintaining the stability and bioactivity of the macromolecular proteins when the device is administered to an animal.
Summary of the Invention
It is, therefore, an object of the present invention to provide a sustained release composition for macromolecular proteins.
It is another object of the present invention to provide a sustained release composition for macromolecular proteins which controls the manner and timing of delivery while maintaining the stability and bioactivity of the macromolecular proteins.
These and other objects are achieved using a sustained release composition for macromolecular proteins which comprises a solid wax matrix having the macromolecular protein and a water-insoluble surfactant uniformly dispersed therein. The water-insoluble surfactant provides advantageous sustained release properties for the protein; the protein is released from the composition in amounts greater than amounts released from protein-wax compositions or from protein-wax compositions containing a water-soluble surfactant. The total amount of protein released from the present composition is greater than the total amount of protein released from protei -wax compositions or from protein-wax compositions containing a water-soluble surfactant and the amount of protein released at any particular time following administration is greater than the amount of protein released from protein-wax compositions or from protein-wax compositions containing a water-soluble surfactant.
Other objects, advantages, and novel features of the present invention will become apparent from the following detailed description of the invention.
Description of the Drawings
Figure 1 shows the cumulative percent protein released for different weight percents of beeswax compositions containing Mazol®.
Figure 2 shows the cumulative percent protein released for different weight percents of beeswax compositions containing polypropylene glycol (PPG) .
Figure 3 shows the cumulative percent protein released for different weight percents of beeswax compositions alone, containing PPG, and containing pentaerytherol. Figure 4 shows the cumulative percent protein released for different surfactants having different Hydrophile Lipophile Balance's (HLBs).
Figure 5 shows the cumulative percent lysozyme released for different Mazol® concentrations.
Figure 6 shows the cumulative percent Bovine Serum Albumin (BSA) released for different waxes.
Figure 7 shows the cumulative percent protein released for different formulation techniques.
Detailed Description of the Invention
The present invention is a sustained release composition for delivering macromolecular proteins to an animal over a prolonged period which comprises a solid wax matrix having the macromolecular protein and a water-insoluble surfactant uniformly dispersed therein. Surprisingly, the inclusion of a water-insoluble surfactant in a wax-protein composition provides advantageous sustained release properties for the protein; the protein is released from the composition in amounts greater than amounts released from protein-wax compositions or from protein-wax compositions containing a water-soluble surfactant. The total amount of protein released from the present composition is greater than the total amount of protein released from protein-wax compositions or from protein-wax compositions containing a water-soluble surfactant, and the amount of protein released at any particular time following administration is greater than the amount of protein released from protein-wax compositions or from protein-wax compositions containing a water-soluble surfactant.
The composition of the present invention can be formed by any procedure (1) which provides an essentially uniform solid wax matrix containing the macromolecular protein and water-insoluble surfactant and (2) which is non-destructive to the macromolecular protein. Several such methods are known in the art. In one preferred embodiment, (1) the wax is heated until it melts; (2) the water-insoluble surfactant is added to the melt and the surfactant and wax composition are mixed thoroughly until the surfactant is uniformly dispersed in the wax composition; and (3) the macromolecular protein is added to the melt and the protein and wax are mixed thoroughly until the protein is uniformly dispersed in the wax. The order of addition of the protein and surfactant is not crucial. In another embodiment, the dry particles of the wax, macromolecular protein, and water-insoluble surfactant are mixed thoroughly to produce a homogeneous mixture. The mixture is subsequently compressed or preferably molded into the desired size and shape.
Waxes useful for producing the composition include animal waxes such as beeswax, lanolin, shellac wax, and Chinese insect wax; vegetable waxes such as hydrogenated soybean oil, cottonseed oil, carnauba, candelilla, bayberry, and sugar cane; and mineral waxes such as fossil or earth waxes (ozocerite, ceresin, montan) and petroleum waxes (paraffin, microcrystalline, slack or scale wax), or combinations thereof. Preferably, the wax material used in the present invention is beeswax, vegetable wax, carnauba wax, or combinations thereof.
The wax or combination of waxes should comprise from about 50-99% by weight of the composition, preferably from about 70-99%. Macromolecular proteins suitable for inclusion and thus deliverable in the sustained release composition of the present invention include but are not limited to proteins having a molecular weight from about 2000 to about 200,000 daltons. The macromolecular proteins include but are not limited to natural, recombinant, synthetic and mutein proteins which have deleted, inserted, substituted, or otherwise modified sequences and biologically active fragments and derivatives thereof. More specifically, bioactive proteins such as enzymes, enzyme inhibitors, antibodies, antigens, interferons, insulins, prolactins, somatomedins, somatostatins, interleukins, somatocrinins (GRF) and somatotropins can be delivered according to the present invention.
Preferably, human, porcine, bovine, equine, ovine, and avian somatotropins can be delivered using the sustained release compositions of the present invention. Somatotropin is defined herein to include all proteins having somatotropin activity including the natural, recombinant, synthetic, and mutein somatotropins having deleted, inserted, substituted, or otherwise modified sequences and biologically active fragments and derivatives thereof. Additionally, metals or metal compounds associated with biologically active proteins, peptides and polypeptides, as well as acid salts, derivatives and complexes and antihydrating agents are suitable for incorporation into the sustained release composition of the invention. Somatotropins useful in the present invention can be obtained from any suitable source. Methods for producing, isolating and purifying native and recombinant somatotropins are well known in the art. The amino acid sequences of various somatotropins useful in the present invention are well known: C.H. Li in Kirk-Othmer "Encyclopedia of Chemical Technology", 3rd E., Vol. 12, pp. 549-552 (human somatotropin), R.P. Woychik, Nucleic Acid Res. , Vol. 10, p. 7197 (1982) (bovine somatotropin), C.H. Li et ah., Arch. Biochem. Biophvs.. Vol. 156, p. 493-508 (1973) (ovine somatotropin) and P.H. Seeburg et al. , DNA, Vol. 2, p. 37, 45 (1983) (porcine somatotropin).
Recombinant somatotropins are also known in the art. European Patent Application Publication No. 0 103 395 describes the construction of a transformant strain of E__ coli containing a first plasmid which codes for delta-9 (Ser) bovine somatotropin (somatotropin less its 9 N-terminal amino acids and having an additional serine residue at the N- erminus) under the control of the lambda PL promoter-operator and which has a Shine-Dalgarno region derived from bacteriophage mu. The transformant also contains a second plasmid, pcI857, which codes for the production of the pcI857 temperature-sensitive repressor protein. The repressor protein can be inactivated by raising the temperature to about 42°C, thereby inducing expression of delta-9 (Ser) bovine somatotropin. A transformant strain of this type, E^ coli HB101 (PL-mu-delta-9 (Ser) bovine somatotropin and pcl857) has been deposited with The
American Type Culture Collection (ATCC), Rockville, MD, and assigned Accession No. 53030.
Construction of a similar transformant strain which codes for the production of delta-7 porcine somatotropin (porcine somatotropin less its first 7
N-terminal amino acids) is described in European Patent Application Publication No. 0 104 920. A transformant strain of this type, E__ coli HB101 (PL-mu-delta-7 porcine somatotropin and pcI857) has been deposited with ATCC and assigned Accession No. 53031.
Strains 53030 and 53031 are prolific producers of delta-9 (Ser) bovine somatotropin and delta-7 porcine somatotropin, respectively. Other methods for many similar proteins are known in the art.
The macromolecular proteins should comprise about 1-30% by weight of the composition, preferably from about 5-20%. Water-insoluble surfactants suitable for inclusion in the sustained release composition of the present invention include non-ionic, water-insoluble surfactants having a molecular weight (MW) of from about 100-2000 and a Hydrophile Lipophile balance (HLB) value of from about 1-17. Such surfactants include glycerol and polyglycerol esters, fatty alcohol ethers, ethoxylated sorbitan fatty acid esters, and the like. Examples of such surfactants, described generally in Table 1, include but are not limited to Mazol®, Macol® CA-2, Macol® CA-20 and T-MAZ®-61. Preferably, the surfactant used in the present invention is Mazol® 80 MG-K. The composition and characteristics of these surfactants are well known to skilled artisans. For example, the surfactants used herein can be purchased from Mazer Chemicals, a division of PPG Industries Chemical Group, Gurnee IL.
The surfactant should comprise from about 0.1-20% by weight of the composition, preferably from about 0.5-10%. In the preferred embodiment of the present invention, the composition comprises from about 50-99% beeswax, 1-30% somatotropin, and 0.1-20% Mazol®.
The composition of the present invention can be produced in many shapes and sizes. The wax melt or dry mixture containing the protein and surfactant can be molded and shaped into any desired form and size. For example, the composition can be sliced into "slabs", shaped into pellets, rods or tablets, and the like. Also, the viscous wax melt can be poured into molds or the dry mixture can be compressed to produce implant devices which can be implanted into an animal.
Pellets, implant devices or tablets preferably comprise from about 50-99% wax, 1-30% protein and 0.1-20% water-insoluble surfactant. Such pellets, implant devices or tablets are designed to deliver the protein to the animal in the required amounts, typically from about 0.1-20 mg protein/animal/day, over a prolonged period, typically from about 1-14 days. Preferably, pellets, implant devices or tablets comprise from about 50-99% beeswax, 1-30% somatotropin and 0.1-20% Mazol®. Such pellets, implant devices or tablets are designed to deliver from about 0.1-20 mg somatotropin/animal/day, preferably from about 1-10 mg/animal/day, over a period of from about 1-14 days. The resulting pellet, implant device or tablet can be implanted into an animal to promote growth and increase feed utilization efficiency.
According to the present invention, a method for delivering macromolecular proteins to an animal comprises administering to the animal the sustained release composition of the present invention.
In another aspect of the present invention, a method for promoting growth and increasing feed utilization efficiency in an animal over a prolonged period comprises administering to an animal the device of the present invention containing somatotropin as the macromolecular protein. The somatotropin should be present in the sustained release composition in an amount effective to promote growth when administered to the animal. Although the amount of somatotropin required to promote growth may vary depending upon the particular somatotropin, the type of animal, and the desired results, the somatotropin is generally present in amounts from about 1-30% by weight of the composition, preferably from about 5-20%. Somatotropin is typically administered to animals in dosages from about 0.1-20 mg/animal/day, preferably from about 1-10 mg/animal/day to promote growth and increase feed utilization efficiency.
The invention having been generally described, the following examples are given as particular embodiments of the invention and to demonstrate the practice and advantages thereof. It is understood that the examples are given by way of illustration and are not intended to limit the specification or the claims to follow in any manner.
Materials used in the Examples The following materials and equipment were used in the examples herein: Bovine Serum Albumin (BSA), lysozyme and micrococcus lysodekticus were obtained from Sigma Chemical Co., St. Louis, MO. Lysozyme was purified, sterilized, and lyophilized before use. Beeswax (white bleached) and candelilla wax (#1 Yellow American) were obtained from Strahl & Pitsch Inc., West Babylon, NY. Stearine (Durkee-07) was obtained from Durkee Industrial Foods, SCM Corporation, Cleveland, Ohio. Surfactants Mazol® 80 MG-K, Macol® CA-2, Macol®
CA-20 and T-MAZ®-61 were obtained from Mazer Chemical Co., Gurnee, IL. The properties of these surfactants are listed in Table 1. Polypropylene glycol (PPG; molecular weight: 425 daltons) was obtained from Aldrich Chemical Co., Milwaukee, WI. Pentaerythritol was sieved to less than 106 microns. A Carver laboratory press (Model #2698 made by Carver Inc., Menomonee Falls, WI. ) was used to prepare tablets.
Release studies were conducted by placing the slabs of tablets in test tubes and shaking them with the buffer solution in a water shaker bath at 37°C. UV absorbance was measured with a Perkin Elmer UV/VIS Spectrophotometer (Model: Lambda 7).
Methods used in the Examples
Sieving of Proteins: BSA was sieved to <106 microns and used in all the experiments except those which studied the effect of particle size on release profile (Trial 7). For Trial 7, BSA was used after sieving to the following fractions: 425-250 microns, 250-150 microns, 150-106 microns, 106-53 microns, and less than 53 microns. Purified lysozyme was sieved to 150-106 microns. Preparation of wax/protein slabs: Beeswax and an appropriate amount of the additive (PPG, Mazol®, etc.) were placed in a 100 ml beaker and heated for about 2 hours in an oven at 150°C with occasional stirring. Five grams of the wax-additive mixture were transferred to a 10 ml beaker and equilibrated for an hour at 85°C in the heating block. The 10 ml beaker containing the wax mixture was then placed on a hot plate stirrer (kept at low setting to minimize heat loss). The protein (0.555 g) was added to the wax mixture, stirred at first with a spatula and then with a magnetic stir bar. The beaker was then placed back into the heating block for about 1 minute to allow wax on the sides of the beaker to remelt. The beaker was placed back onto the hot plate and stirred briefly to resuspend the protein. The magnetic stir bar was removed and the wax/protein suspension was quickly transferred to an aluminum weighing pan (previously warmed on a hot plate). A spatula was used to quickly scrape the wax from the sides of the beaker into the pan. The pan was then transferred to a leveled thermoelectric cold plate set on the coldest setting. After the wax had solidified, the pan was removed from the cold plate and brought to room temperature. The procedure, from the addition of the protein to the wax to the initial solidification of the wax, took about 2 minutes. When pentaerythritol was used as the additive, it was dry-mixed with the solid protein before being added to. the molten wax.
After cooling to room temperature, the slab of protein/wax matrix was removed from the pan and placed onto a glass plate. A rectangular section (2.0 cm x 4 cm) was cut from the center portion of the slab with a razor blade. This was further cut into eight smaller slabs measuring 2.0 cm x 0.5 cm x 0.20-0.25 cm (1 x w x h) . The smaller slabs were weighed individually prior to use in release studies. The loading of the protein was about 10% w/w. Preparation of Compressed Wax Disks: Candelilla wax and Macol® CA-20 (100/2 by weight) were placed into a beaker and melted at 150°C for 1 hour to allow any trapped water to escape. The wax was poured onto a piece of aluminum foil, cooled and ground in a high speed grinding mill in intervals of 15 seconds. The wax powder was sieved after each interval to avoid undersizing the wax. The fraction from 250-150 microns was used. "Dry-mixed" formulations were prepared by mixing BSA with Macol® CA-20 wax powder as prepared above in the ratio of 10/90 (w/w). The matrix (200 mg) was pelleted in a mold (Perkin Elmer; Dia: 1.3 cm) at 24,000 psi in the Carver Press.
"Melt-mixed" formulations were prepared by melting the candelilla wax with the Macol® CA-20 (2%) at 150C for one hour. Five grams of this mixture were placed into a beaker and cooled to 85°C and mixed with BSA (0.555 g) . The suspension was poured onto a piece of aluminum foil to cool and ground in the mill as described previously to collect all the particles <250 microns. The homogeneity of the matrix was checked by sampling out 200 mg of the matrix, extracting it with 10 mM PBS (pH: 7.4), and analyzing the solution for protein concentration. The matrix was then compressed as described before.
"Melt-cast" slabs were prepared with candelilla wax and 2% Macol® CA-20 using the procedure given for the beeswax slabs above. Strips were cut from the slab with a hot razor blade because of the hard, brittle nature of the candelilla wax. The weights and surface areas of the devices from the three techniques were similar. Density of Compressed Disks and Cast Slabs: The true density of the disks were determined with a helium pycnometer. The bulk volumes of the compressed disks were calculated from their dimensions. The bulk volumes of the melt-cast slabs were determined with a pycnometer by displacement of an ethanol/water mixture of known density. The porosities of the compressed disks and cast slabs were calculated from their true volume and bulk volume. In vitro release profiles were conducted on slabs by placing them into screw cap culture tubes (125 mm x 20 mm) which were previously autoclaved, dried, and filled with 8 ml of 10 mM PBS (pH: 7.4). Screw-cap Erlenmeyer flasks (50ml) were used for the compressed disks. Gentamycin sulfate (100 ppm) was added to the buffer as an antimicrobial agent. The tubes were shaken (200 rpm) in a water bath at 37°C. The solutions were decanted and replaced with fresh buffer first after 8 hours and thereafter at 24 hour intervals. Eight implants were used for each system for all studies.
The absorbance of release solutions (previously filtered through 0.22 microns) was measured at 276 nanometers (nm) for BSA and 281 nm for lysozyme. The absorbance at 320 nm was subtracted from the 276 nm and 281 nm values to correct for random scattering. The concentrations of the protein solutions were determined by using the following absorptivities: BSA: 0.595 mg"1 ml cm-1 and lysozyme: 2.41 mg"1 ml cm-1.
Activities of lysozyme samples were analyzed by measuring the change in absorbance of a suspension of micrococcus lysodekticus substrate.
Example 1 Ten (10) trials comprising thirty-one (31) sets of compositions having different combinations of BSA, waxes, and surfactants were evaluated using the materials and methods described above. The compositions are shown in Table 2. The results are shown in Figures 1 and 2.
Referring to Figures 1 and 2, the water-insoluble surfactant Mazol® dramatically increased the amount of protein released from beeswax pellets when compared to the water-soluble surfactant PPG. Beeswax-PPG (10%) compositions released only about 25% of the protein after 10 days; beeswax-Mazol (10%) compositions released almost 100% of the protein after 14 days. Indeed, beeswax-Mazol (2%) compositions release about twice as much protein (about 60%) as do the beeswax-PPG (10%) compositions, both compared after 14 days.
Effects of BSA loading and water-soluble additives: The formation of channels/pores in a solid matrix can be enhanced either by increasing the loading of the protein in the matrix or by incorporating other water-soluble additives. In both cases, the transport of the protein is made easier giving rise to high release rates. This point was tested by preparing slabs containing 20% BSA (Set 9) and slabs containing 10% BSA along with 10% pentaerythritol (Set 10) and testing the release profiles as described previously. The results are shown in Figure 3. Referring to Figure 3, both sets 9 and 10 showed similar release profiles and displayed higher release-rates than sets with 10% protein loading (Sets 6 and 11). These data suggest that the release-rates are affected by the type of additive as well as its concentration in the matrix. Comparing Figures 1 and 2, it is clear that high release rates of BSA can be obtained by: (1) increasing the loading of the protein; (2) incorporating large amounts of water soluble additives such as pentaerythritol; and (3) incorporating a small amount of water-insoluble surfactant such as Mazol. The effectiveness of Mazol may be attributed to its water-insoluble, surface-active nature which can promote the permeation of water into the matrix. PPG, on the other hand, is not surface-active, but is water-soluble. The difference between Mazol and PPG was also evident visually during the release study. Slabs containing Mazol became white and slightly swollen, but those containing PPG did not.
Effect of different surfactants as additive: In order to test the effect of different surfactants on the release of BSA, slabs were prepared using surfactants with different HLB (Hydrophile Lipophile balance) numbers. A higher HLB number means the material is more hydrophilic. Table 1 lists the HLB numbers of the different surfactants used. The effectiveness of these additives was compared at a 2% level in the matrix as previously described. The release profiles for these sets are shown in Figure 4.
Referring to Figure 4, there were minor differences in the release profiles for Mazol and Polysorbate 61, but surprisingly there was no difference between Macol® CA-2 and Macol CA-20. This could have arisen from the differences in the molecular weights of Macol CA-2 and Macol CA-20 (about 350 daltons and about 1200 daltons respectively). Thus, the molar concentration of Macol CA-2 in the wax was higher than that of Macol CA-20.
Release of Lysozyme from Wax Slabs: During the preparation of the wax/protein slabs, the protein had to be kept at 85°C for about 2 minutes in the molten ' wax. As these conditions could inactivate the protein, it became necessary to use a protein whose activity could be readily measured. Lysozyme was chosen for this purpose and was released from slabs containing 0.0178 olal Macol CA-2 and 0.0178 molal Macol CA-20. The release profile of the matrix without any surfactant was similar to that with Macol CA-2 (Sets 24 and 25; Figure 5). The slab containing Macol CA-20
(Set 26) released substantially more of the lysozyme. The cumulative amount of lysozyme released, however, was much less than in the case of BSA. At the end of the release study, two slabs from Set 24 were extracted first with PBS and then with 50 mM glycine buffer (pH: 3.0). The total recovery from PBS and glycine buffers was only about 60%, with the balance being unaccounted. In contrast, all the protein could be accounted for in the experiments involving BSA. These results illustrate the intrinsic differences between these two proteins.
Release studies with lysozyme were stopped after 20 days and the residual protein was extracted from the wax slabs for measurement of bioactivity. Activity assays were performed on the following four lysozyme samples: (i) as received; (ii) purified; (iii) lysozyme extracted from a freshly cast slab and (iv) lysozyme recovered from the slabs at the end of the release study. The results are shown in Table 3. Referring to Table 3, there was no difference in the activity of the four samples indicating that neither the formulation technique nor the release conditions had affected the bioactivity.
Example 5 A device containing BSA in a 1:1 mixture of beeswax and Stearine was formulated for testing as described previously. This composition also contained Macol CA-20 at 2% by weight (0.0178 molal). The composition was tested as previously described for release characteristics. The results are shown graphically in Figure 6.
Referring to Figure 6, release of BSA was more rapid from the Stearine/beeswax matrix than from the beeswax matrix (Set 20).
Comparison of Cast Wax Slabs and Compressed Wax Disks: As discussed in the Background, there is prior art relating to the sustained release of low molecular weight organic drugs from wax matrices. In almost all the cases cited, the drug was dispersed or dissolved in molten wax. The matrix was then crushed, sized, and tableted. Sustained release of the drug was obtained from these devices over a period of several hours (usually less than 24 hours). The presence of surfactants in the matrix influenced the release rate of the drug. Other significant variables were the particle size before tableting and the drug loading. The role of the formulation technique in prolonging the duration of release was determined. Three techniques were compared in their effectiveness to achieve sustained release of BSA: (a) "Melt-cast slab" (Set 29); (b) "Dry-mixed compressed disk" (Set 30), and (c) "Melt-mixed compressed disk" (Set 31).
Compressed disks and cast slabs were prepared as previously described and tested for release profile. Candelilla wax was used in all the cases because of its low melting point (65-70°C) and ease of tableting. The surface areas of the disks and the slabs were similar (3.31 sq. cm vs 3.25 sq. cm). The loading of BSA was 10% w/w for all disks and slabs tested. The porosities of the compressed disks and cast-slabs were calculated using the equation: E = 100(l-(vt/vb) ) , where vt = true volume and vb = bulk volume of the matrix, respectively. The true volume and bulk volume were determined as described previously. The results are shown in Figure 7.
Referring to Figure 7, release of BSA was much faster from the compressed disks than from the cast slab. The "dry-mixed" and "melt-mixed" compressed disks (Sets 30 and 31 respectively) showed similar release profiles and released 100% in about 3 days.
The "melt-mixed" disks, however, gave a slower release rate than the "dry-mixed" disks in the first 24 hours of the study. In the "melt-mixed" technique, the protein was added to the molten wax.
Example 7 A mixture of Zn-rpST/L-Arginine (1:1) was formulated into slabs containing Beeswax (100%) or Beeswax/Mazol (90:10). The compositions were prepared and tested as described above. From the results shown in Table 4, it is evident that PST release is higher from wax slabs containing Mazol than from those without Mazol.
Obviously many modifications and variations of the present invention are possible in light of the above teachings. It is therefore to be understood that within the scope of the appended claims the invention may be practiced otherwise than as specifically described. Table 1 Properties of Surfactants Used
Surfactant Chemical Classification Molecular HLB2
Mazol® 80 MG-K Ethoxylated mono- and 1400 13.5 di-glycerides Macol® CA-2 Polyoxyethylene ether 30 4.9 of cetyl alcohol (2 units of EO)
Macol® CA-20 Polyoxyethylene ether 1122 15.7 of cetyl alcohol (20 units of EO)
T-MAZ®-61 Polyoxyethylene ether 706 9.5 of sorbitan monostearate (4 units of EO)
Hydrophile Lipophile Balance number EO = ethylene oxide
Table 2 Compositions of the Systems Evaluated
Set Code Trial Description
Beeswax/Macol CA2/BSA 90/2/10
Beeswax/Macol CA20/BSA 90/2/10
Beeswa /Lysozyme 90/10
Beeswax/Macol CA2/Lysozyme 90/.59/10
Beeswax/Macol CA20/Lysozyme 90/2/10 Stearin/Beeswax/CA-20/BSA 45/45/2/10 Candelilla Wax/CA-20/BSA 90/2/10 Melt cast slab Candelilla Wax/CA-20/BSA 90/2/10 Compressed disd; dry mixed
Activity of Lysozyme Recovered From the Wax Formulations
Sample Activity (10" 4*).1
1 units/mg protein (Average of 5 replicates) purified lysozyme used in slabs extracted from unreleased slab one sample from each set was used
Release of PST from Beeswax Slabs with and without Mazol
Time Cumulative Percent Released (Days) Beeswax without Mazol Beeswax with Mazol
|Cited Patent||Filing date||Publication date||Applicant||Title|
|EP0246540A2 *||12 May 1987||25 Nov 1987||Paul Y. Wang||Implant preparations for delivery of bioactive macromolecules|
|EP0257368A1 *||4 Aug 1987||2 Mar 1988||American Cyanamid Company||Compositions for parenteral administration and their use|
|US2413419 *||27 Feb 1943||31 Dec 1946||Searle & Co||Pellet for administering gonadotropic pituitary hormones|
|Citing Patent||Filing date||Publication date||Applicant||Title|
|WO1993013792A1 *||8 Jan 1993||22 Jul 1993||Pitman-Moore, Inc.||Delayed release device for transition metal/protein complexes|
|WO2004083231A2 *||11 Mar 2004||30 Sep 2004||Centre National De La Recherche Scientifique||Use of peptide ligands for plasma membrane receptors or similar for mitochondrial activity modulation|
|WO2004083231A3 *||11 Mar 2004||11 Nov 2004||Centre Nat Rech Scient||Use of peptide ligands for plasma membrane receptors or similar for mitochondrial activity modulation|
|WO2004091510A2||12 Apr 2004||28 Oct 2004||Medimmune, Inc.||Recombinant il-9 antibodies and uses thereof|
|WO2006102095A2||17 Mar 2006||28 Sep 2006||Medimmune, Inc.||Framework-shuffling of antibodies|
|WO2007002543A2||23 Jun 2006||4 Jan 2007||Medimmune, Inc.||Antibody formulations having optimized aggregation and fragmentation profiles|
|WO2007024715A2||18 Aug 2006||1 Mar 2007||Abbott Laboratories||Dual variable domain immunoglobin and uses thereof|
|WO2007147090A2||14 Jun 2007||21 Dec 2007||Macrogenics, Inc.||Methods for the treatment of autoimmune disorders using monoclonal antibodies with reduced toxicity|
|WO2008157379A2||13 Jun 2008||24 Dec 2008||Macrogenics, Inc.||Covalent diabodies and uses thereof|
|WO2009092011A1||16 Jan 2009||23 Jul 2009||Medimmune, Llc||Cysteine engineered antibodies for site-specific conjugation|
|WO2009149185A2||3 Jun 2009||10 Dec 2009||Abbott Laboratories||Dual variable domain immunoglobulins and uses thereof|
|WO2010080538A1||17 Dec 2009||15 Jul 2010||Macrogenics, Inc.||Covalent diabodies and uses thereof|
|WO2010095130A1 *||17 Feb 2010||26 Aug 2010||Technion - Research & Development Foundation Ltd||Food containing prolactin|
|WO2010099079A1||23 Feb 2010||2 Sep 2010||Abbott Laboratories||Antibodies to troponin i and methods of use thereof|
|WO2010104815A1||9 Mar 2010||16 Sep 2010||Abbott Laboratories||Antibodies binding to pivka-ii amino acids 13-27|
|WO2010141329A1||28 May 2010||9 Dec 2010||Medimmune, Llc||Molecules with extended half-lives and uses thereof|
|WO2011025964A2||27 Aug 2010||3 Mar 2011||Abbott Laboratories||Therapeutic dll4 binding proteins|
|WO2011044368A1||7 Oct 2010||14 Apr 2011||Macrogenics, Inc.||Fc region-containing polypeptides that exhibit improved effector function due to alterations of the extent of fucosylation, and methods for their use|
|WO2011070045A1||8 Dec 2010||16 Jun 2011||Abbott Gmbh & Co. Kg||Monoclonal antibodies against the rgm a protein for use in the treatment of retinal nerve fiber layer degeneration|
|WO2011130377A2||13 Apr 2011||20 Oct 2011||Abbott Laboratories||Amyloid-beta binding proteins|
|WO2011138391A1||5 May 2011||10 Nov 2011||Novartis Ag||Compositions and methods of use for therapeutic low density lipoprotein - related protein 6 (lrp6) multivalent antibodies|
|WO2011138392A1||5 May 2011||10 Nov 2011||Novartis Ag||Compositions and methods of use for therapeutic low density lipoprotein -related protein 6 (lrp6) antibodies|
|WO2011143562A2||13 May 2011||17 Nov 2011||Abbott Laboratories||Il-1 binding proteins|
|WO2012006500A2||8 Jul 2011||12 Jan 2012||Abbott Laboratories||Monoclonal antibodies against hepatitis c virus core protein|
|WO2012018476A1||8 Jul 2011||9 Feb 2012||Abbott Laboratories||Antibodies relating to pivka-ii and uses thereof|
|WO2012018687A1||29 Jul 2011||9 Feb 2012||Macrogenics, Inc.||Covalent diabodies and uses thereof|
|WO2012022814A1||22 Aug 2011||23 Feb 2012||Novartis Ag||Antibodies for epidermal growth factor receptor 3 (her3)|
|WO2012024187A1||12 Aug 2011||23 Feb 2012||Abbott Laboratories||Amyloid-beta binding proteins|
|WO2012024650A2||19 Aug 2011||23 Feb 2012||Abbott Laboratories||Anti-ngf antibodies and their use|
|WO2012069466A1||22 Nov 2011||31 May 2012||Novartis Ag||Multispecific molecules|
|WO2012088094A2||20 Dec 2011||28 Jun 2012||Abbott Laboratories||Il-1 binding proteins|
|WO2012121775A2||20 Dec 2011||13 Sep 2012||Abbott Laboratories||Dual variable domain immunoglobulins and uses thereof|
|WO2012145493A1||19 Apr 2012||26 Oct 2012||Amplimmune, Inc.||Antibodies and other molecules that bind b7-h1 and pd-1|
|WO2012162068A2||16 May 2012||29 Nov 2012||Macrogenics, Inc.||Deimmunized serum-binding domains and their use for extending serum half-life|
|WO2013025779A1||15 Aug 2012||21 Feb 2013||Amplimmune, Inc.||Anti-b7-h4 antibodies and their uses|
|WO2013043070A2||19 Sep 2012||28 Mar 2013||Technophage, Investigação E Desenvolvimento Em Biotecnologia, Sa||Anti-tumor necrosis factor-alpha agents and uses thereof|
|WO2013063095A1||24 Oct 2012||2 May 2013||Abbvie Inc.||Immunobinders directed against sclerostin|
|WO2013084147A2||4 Dec 2012||13 Jun 2013||Novartis Ag||Antibodies for epidermal growth factor receptor 3 (her3)|
|WO2013084148A2||4 Dec 2012||13 Jun 2013||Novartis Ag||Antibodies for epidermal growth factor receptor 3 (her3) directed to domain ii of her3|
|WO2013090633A2||13 Dec 2012||20 Jun 2013||AbbVie Deutschland GmbH & Co. KG||Composition and method for the diagnosis and treatment of iron-related disorders|
|WO2013090635A2||13 Dec 2012||20 Jun 2013||AbbVie Deutschland GmbH & Co. KG||Composition and method for the diagnosis and treatment of iron-related disorders|
|WO2013093809A1||19 Dec 2012||27 Jun 2013||Pfizer Inc.||Engineered antibody constant regions for site-specific conjugation and methods and uses therefor|
|WO2013112922A1||25 Jan 2013||1 Aug 2013||AbbVie Deutschland GmbH & Co. KG||Composition and method for diagnosis and treatment of diseases associated with neurite degeneration|
|WO2013184514A1||31 May 2013||12 Dec 2013||Irm Llc||Site-specific labeling methods and molecules produced thereby|
|WO2014011955A2||12 Jul 2013||16 Jan 2014||Abbvie, Inc.||Il-1 binding proteins|
|WO2014072876A1||28 Oct 2013||15 May 2014||Pfizer Inc.||Platelet-derived growth factor b specific antibodies and compositions and uses thereof|
|WO2014100483A1||19 Dec 2013||26 Jun 2014||Amplimmune, Inc.||Anti-human b7-h4 antibodies and their uses|
|WO2014100823A1||23 Dec 2013||26 Jun 2014||Amplimmune, Inc.||Anti-h7cr antibodies|
|WO2014124258A2||7 Feb 2014||14 Aug 2014||Irm Llc||Specific sites for modifying antibodies to make immunoconjugates|
|WO2014124316A2||7 Feb 2014||14 Aug 2014||Irm Llc||Specific sites for modifying antibodies to make immunoconjugates|
|WO2014144355A2||14 Mar 2014||18 Sep 2014||Abbott Laboratories||Anti-gp73 monoclonal antibodies and methods of obtaining the same|
|WO2014159239A2||10 Mar 2014||2 Oct 2014||Novartis Ag||Antibodies against notch 3|
|WO2014160160A2||13 Mar 2014||2 Oct 2014||Novartis Ag||Antibody drug conjugates|
|WO2014190356A2||27 May 2014||27 Nov 2014||Amplimmune, Inc.||Anti-b7-h5 antibodies and their uses|
|WO2015058868A1||11 Jun 2014||30 Apr 2015||Pangaea Biotech, S.L.||Compositions and methods for the treatment of cancer|
|WO2015073580A1||12 Nov 2014||21 May 2015||Pfizer Inc.||Tumor necrosis factor-like ligand 1a specific antibodies and compositions and uses thereof|
|WO2015084883A2||2 Dec 2014||11 Jun 2015||Abbvie, Inc.||Compositions and methods for treating osteoarthritis|
|WO2015109212A1||16 Jan 2015||23 Jul 2015||Pfizer Inc.||Anti-il-2 antibodies and compositions and uses thereof|
|WO2015138337A1||9 Mar 2015||17 Sep 2015||Abbvie, Inc.||Compositions and methods for treating rheumatoid arthritis|
|WO2015138615A2||11 Mar 2015||17 Sep 2015||Irm Llc||Specific sites for modifying antibodies to make immunoconjugates|
|WO2016020791A1||27 Jul 2015||11 Feb 2016||Novartis Ag||Ckit antibody drug conjugates|
|WO2016024195A1||7 Aug 2015||18 Feb 2016||Novartis Ag||Anti-cdh6 antibody drug conjugates|
|WO2016075670A1||13 Nov 2015||19 May 2016||Novartis Ag||Antibody drug conjugates|
|WO2016094881A2||11 Dec 2015||16 Jun 2016||Abbvie Inc.||Lrp-8 binding proteins|
|WO2016118921A1||22 Jan 2016||28 Jul 2016||Abbvie, Inc.||Compositions and methods for treating psoriatic arthritis|
|WO2016139482A1||3 Mar 2016||9 Sep 2016||Kymab Limited||Antibodies, uses & methods|
|WO2016203432A1||17 Jun 2016||22 Dec 2016||Novartis Ag||Antibody drug conjugates|
|WO2017046676A1||8 Sep 2016||23 Mar 2017||Novartis Ag||Polyomavirus neutralizing antibodies|
|WO2017070561A1||21 Oct 2016||27 Apr 2017||Pfizer Inc.||Anti-il-2 antibodies and compositions and uses thereof|
|WO2017096051A1||1 Dec 2016||8 Jun 2017||Stcube & Co., Inc.||Antibodies and molecules that immunospecifically bind to btn1a1 and the therapeutic uses thereof|
|WO2017125897A1||20 Jan 2017||27 Jul 2017||Novartis Ag||Multispecific molecules targeting cll-1|
|EP1104296A1 *||20 Jul 1999||6 Jun 2001||Peptech Limited||Bioimplant formulation|
|EP1104296B1 *||20 Jul 1999||13 Jun 2012||Peptech Animal Health Pty Limited||Bioimplant formulation|
|EP1420818A1 *||26 Jul 2002||26 May 2004||Agresearch Limited||Antigenic compositions|
|EP2027874A2||28 Nov 2001||25 Feb 2009||Medimmune, Inc.||Methods of administering/dosing anti-rsv antibodies for prophylaxis and treatment|
|EP2116556A1||8 May 2009||11 Nov 2009||Abbott GmbH & Co. KG||Antibodies to receptor of advanced glycation end products (rage) and uses thereof|
|EP2272530A2 *||26 Jul 2002||12 Jan 2011||Otago Innovation Limited||Antigenic Compositions|
|EP2272530A3 *||26 Jul 2002||22 May 2013||Otago Innovation Limited||Antigenic Compositions|
|EP2272533A1||13 Jan 2004||12 Jan 2011||MacroGenics, Inc.||Soluble FcyR fusion proteins and methods of use thereof|
|EP2272566A2||18 Aug 2004||12 Jan 2011||MedImmune, LLC||Humanisation of antibodies|
|EP2292663A2||24 Aug 2007||9 Mar 2011||Kyowa Hakko Kirin Co., Ltd.||Antagonistic human light-specific human monoclonal antibodies|
|EP2316487A1||12 Apr 2004||4 May 2011||MedImmune, LLC||Recombinant IL-9 antibodies & uses thereof|
|EP2338512A1||28 Nov 2001||29 Jun 2011||MedImmune, LLC||Methods of administering/dosing anti-RSV antibodies for prophylaxis and treatment|
|EP2341060A1||12 Dec 2001||6 Jul 2011||MedImmune, LLC||Molecules with extended half-lives, compositions and uses thereof|
|EP2354149A1||12 Dec 2001||10 Aug 2011||MedImmune, LLC||Molecules with extended half-lives, compositions and uses thereof|
|EP2357187A1||12 Dec 2001||17 Aug 2011||MedImmune, LLC||Molecules with extended half-lives, compositions and uses thereof|
|EP2368578A1||9 Jan 2004||28 Sep 2011||Macrogenics, Inc.||Identification and engineering of antibodies with variant Fc regions and methods of using same|
|EP2371389A2||14 Aug 2003||5 Oct 2011||MacroGenics, Inc.||FcgammaRIIB-specific antibodies and methods of use thereof|
|EP2407548A1||16 Oct 2007||18 Jan 2012||MedImmune, LLC||Molecules with reduced half-lives, compositions and uses thereof|
|EP2412384A1||28 Nov 2001||1 Feb 2012||MedImmune, LLC||Methods of administering/dosing anti-RSV antibodies for prophylaxis and treatment|
|EP2422811A2||27 Oct 2005||29 Feb 2012||MedImmune, LLC||Modulation of antibody specificity by tailoring the affinity to cognate antigens|
|EP2484696A1||24 Aug 2007||8 Aug 2012||Kyowa Hakko Kirin Co., Ltd.||Antagonistic human light-specific human monoclonal antibodies|
|EP2495257A2||18 Aug 2006||5 Sep 2012||Abbott Laboratories||Dual variable domain immunoglobulin and uses thereof|
|EP2500352A1||18 Aug 2006||19 Sep 2012||Abbott Laboratories||Dual variable domain immunoglobulin and uses thereof|
|EP2500353A2||18 Aug 2006||19 Sep 2012||Abbott Laboratories||Dual variable domain immunoglobulin and uses thereof|
|EP2500354A2||18 Aug 2006||19 Sep 2012||Abbott Laboratories||Dual variable domain immunoglobulin and uses thereof|
|EP2500355A2||18 Aug 2006||19 Sep 2012||Abbott Laboratories||Dual variable domain immunoglobulin and uses thereof|
|EP2500356A2||18 Aug 2006||19 Sep 2012||Abbott Laboratories||Dual variable domain immunoglobulin and uses thereof|
|EP2500357A2||18 Aug 2006||19 Sep 2012||Abbott Laboratories||Dual variable domain immunoglobulin and uses thereof|
|EP2500358A2||18 Aug 2006||19 Sep 2012||Abbott Laboratories||Dual variable domain immunoglobulin and uses thereof|
|EP2500359A2||18 Aug 2006||19 Sep 2012||Abbott Laboratories||Dual variable domain immunoglobulin and uses thereof|
|EP2500361A1||8 May 2009||19 Sep 2012||Abbott GmbH & Co. KG||Antibodies to receptor of advanced glycation end products (rage) and uses thereof|
|EP2505209A1||26 Jun 2007||3 Oct 2012||MacroGenics, Inc.||Fcgamma-RIIB-specific antibodies and methods of the use thereof|
|EP2520588A1||18 Aug 2006||7 Nov 2012||Abbott Laboratories||Dual variable domain immunoglobulin and uses thereof|
|EP2573114A1||10 Aug 2006||27 Mar 2013||MacroGenics, Inc.||Identification and engineering of antibodies with variant Fc regions and methods of using same|
|EP2626371A1||30 Jul 2008||14 Aug 2013||MedImmune, LLC||Multispecific epitope binding proteins and uses thereof|
|EP2711018A1||21 Jun 2010||26 Mar 2014||MedImmune, LLC||Engineered Fc regions for site-specific conjugation|
|EP2772269A2||5 Mar 2010||3 Sep 2014||Abbvie Inc.||IL-17 binding proteins|
|EP2786762A2||17 Dec 2009||8 Oct 2014||MacroGenics, Inc.||Covalent diabodies and uses thereof|
|EP2810652A2||5 Mar 2010||10 Dec 2014||AbbVie Inc.||IL-17 binding proteins|
|EP2810654A1||8 Jul 2009||10 Dec 2014||AbbVie Inc.||Prostaglandin E2 binding proteins and uses thereof|
|EP2815764A1||14 Jun 2007||24 Dec 2014||Macrogenics, Inc.||Methods for the treatment of autoimmune disorders using monoclonal antibodies with reduced toxicity|
|EP2839842A1||20 Dec 2013||25 Feb 2015||MacroGenics, Inc.||Bi-specific monovalent diabodies that are capable of binding CD123 and CD3 and uses thereof|
|EP2840091A1||20 Dec 2013||25 Feb 2015||MacroGenics, Inc.||Bi-specific diabodies that are capable of binding gpA33 and CD3 and uses thereof|
|EP2853540A1||23 Feb 2010||1 Apr 2015||Abbott Laboratories||Antibodies to troponin I and methods of use therof|
|EP2899209A1||28 Apr 2009||29 Jul 2015||Abbvie Inc.||Dual Variable Domain Immunoglobulins and uses thereof|
|EP2921177A2||8 Jul 2011||23 Sep 2015||AbbVie Inc.||Dual variable domain immunoglobulins and uses thereof|
|EP3002299A1||3 Jun 2009||6 Apr 2016||AbbVie Inc.||Dual variable domain immunoglobulins and uses thereof|
|EP3029070A1||27 Aug 2010||8 Jun 2016||AbbVie Inc.||Therapeutic dll4 binding proteins|
|EP3059248A1||8 May 2009||24 Aug 2016||Abbvie Deutschland GmbH & Co. KG||Antibodies to receptor of advanced glycation end products (rage) and uses thereof|
|EP3059249A1||9 Mar 2010||24 Aug 2016||Abbott Laboratories||Antibodies binding to pivka-ii amino acids 13-27|
|EP3072904A1||28 Feb 2011||28 Sep 2016||Abbvie Inc.||Therapeutic dll4 binding proteins|
|EP3124499A1||14 Mar 2014||1 Feb 2017||Abbott Laboratories||Anti-gp73 monoclonal antibodies and methods of obtaining the same|
|US5662917 *||22 Mar 1996||2 Sep 1997||Lucky Limited||Release-controlled implantable somatotropin composition|
|US6818216||28 Nov 2001||16 Nov 2004||Medimmune, Inc.||Anti-RSV antibodies|
|US6855493||28 Nov 2001||15 Feb 2005||Medimmune, Inc.||Methods of administering/dosing anti-RSV antibodies for prophylaxis and treatment|
|US7132100||13 Jun 2003||7 Nov 2006||Medimmune, Inc.||Stabilized liquid anti-RSV antibody formulations|
|US7179900||31 Mar 2003||20 Feb 2007||Medimmune, Inc.||Methods of administering/dosing anti-RSV antibodies for prophylaxis and treatment|
|US7229619||28 Nov 2000||12 Jun 2007||Medimmune, Inc.||Methods of administering/dosing anti-RSV antibodies for prophylaxis and treatment|
|US7294336||24 Feb 2006||13 Nov 2007||Medimmune, Inc.||Stabilized liquid anti-RSV antibody formulations|
|US7323172||8 Oct 2004||29 Jan 2008||Medimmune, Inc.||Methods of administering/dosing anti-RSV antibodies for prophylaxis and treatment|
|US7326571||17 Jul 2003||5 Feb 2008||Boston Scientific Scimed, Inc.||Decellularized bone marrow extracellular matrix|
|US7425618||13 Jun 2003||16 Sep 2008||Medimmune, Inc.||Stabilized anti-respiratory syncytial virus (RSV) antibody formulations|
|US7553489||20 Dec 2006||30 Jun 2009||Medimmune, Llc||Methods of administering/dosing anti-RSV antibodies for prophylaxis and treatment|
|US7612181||18 Aug 2006||3 Nov 2009||Abbott Laboratories||Dual variable domain immunoglobulin and uses thereof|
|US7700739||29 Jun 2006||20 Apr 2010||Abbott Laboratories||IL-12/p40 binding proteins|
|US7915388||7 Sep 2007||29 Mar 2011||Abbott Laboratories||Interleukin-13 binding proteins|
|US8030026||24 Feb 2009||4 Oct 2011||Abbott Laboratories||Antibodies to troponin I and methods of use thereof|
|US8052982||27 Mar 2007||8 Nov 2011||Peptech Animal Health Pty Limited||Bioimplant formulation comprising lecithin and stearin|
|US8283162||10 Mar 2009||9 Oct 2012||Abbott Laboratories||Antibodies relating to PIVKAII and uses thereof|
|US8323651||8 May 2009||4 Dec 2012||Abbott Laboratories||Antibodies to receptor of advanced glycation end products (RAGE) and uses thereof|
|US8383778||29 Jan 2010||26 Feb 2013||Abbvie Inc.||IL-1 binding proteins|
|US8420083||28 Oct 2010||16 Apr 2013||Abbvie Inc.||Antibodies to receptor for advanced glycation end products (RAGE) and uses thereof|
|US8586714||1 Sep 2010||19 Nov 2013||Abbvie, Inc.||Dual variable domain immunoglobulins and uses thereof|
|US8604177||8 Mar 2011||10 Dec 2013||Abbvie Inc.||Interleukin-13 binding proteins|
|US8623358||27 Aug 2010||7 Jan 2014||Abbvie Inc.||Therapeutic DLL4 binding proteins|
|US8629257||19 Apr 2010||14 Jan 2014||Abbvie Inc.||IL-12/p40 binding proteins|
|US8691224||30 Nov 2006||8 Apr 2014||Abbvie Inc.||Anti-Aβ globulomer 5F7 antibodies|
|US8716450||15 Oct 2010||6 May 2014||Abbvie Inc.||Dual variable domain immunoglobulins and uses thereof|
|US8722855||28 Oct 2010||13 May 2014||Abbvie Inc.||Dual variable domain immunoglobulins and uses thereof|
|US8735546||2 Aug 2011||27 May 2014||Abbvie Inc.||Dual variable domain immunoglobulins and uses thereof|
|US8790920||29 Jan 2008||29 Jul 2014||Boston Scientific Scimed, Inc.||Decellularized bone marrow extracellular matrix|
|US8822645||8 Jul 2009||2 Sep 2014||Abbvie Inc.||Prostaglandin E2 dual variable domain immunoglobulins and uses thereof|
|US8835610||5 Mar 2010||16 Sep 2014||Abbvie Inc.||IL-17 binding proteins|
|US8841417||13 May 2011||23 Sep 2014||Abbvie Inc.||IL-1 binding proteins|
|US8877190||6 May 2011||4 Nov 2014||Abbvie Inc.||Aβ conformer selective anti-Aβ globulomer monoclonal antibodies|
|US8895004||27 Feb 2008||25 Nov 2014||AbbVie Deutschland GmbH & Co. KG||Method for the treatment of amyloidoses|
|US8906864||29 Sep 2006||9 Dec 2014||AbbVie Deutschland GmbH & Co. KG||Binding domains of proteins of the repulsive guidance molecule (RGM) protein family and functional fragments thereof, and their use|
|US8916517||2 Nov 2010||23 Dec 2014||The Administrators Of The Tulane Educational Fund||Analogs of pituitary adenylate cyclase-activating polypeptide (PACAP) and methods for their use|
|US8962803||20 Feb 2009||24 Feb 2015||AbbVie Deutschland GmbH & Co. KG||Antibodies against the RGM A protein and uses thereof|
|US8986686||30 Apr 2013||24 Mar 2015||Medimmune, Llc||Stabilized liquid anti-RSV antibody formulations|
|US8987418||3 Jul 2014||24 Mar 2015||Abbvie Inc.||Dual specific binding proteins directed against IL-1β and/or IL-17|
|US8987419||13 Apr 2011||24 Mar 2015||AbbVie Deutschland GmbH & Co. KG||Amyloid-beta binding proteins|
|US9006181||21 Jul 2005||14 Apr 2015||The Administrators Of The Tulane Educational Fund||Treatment of renal dysfunction and multiple myeloma using PACAP compounds|
|US9029508||28 Apr 2009||12 May 2015||Abbvie Inc.||Dual variable domain immunoglobulins and uses thereof|
|US9035027||3 Jun 2009||19 May 2015||Abbvie Inc.||Dual variable domain immunoglobulins and uses thereof|
|US9045551||11 Jun 2014||2 Jun 2015||Abbvie Inc.||Anti-DLL4/VEGF dual variable domain immunoglobulin and uses thereof|
|US9046513||25 Aug 2011||2 Jun 2015||Abbvie Inc.||Dual variable domain immunoglobulins and uses thereof|
|US9062101||12 Aug 2011||23 Jun 2015||AbbVie Deutschland GmbH & Co. KG||Amyloid-beta binding proteins|
|US9062108||14 Mar 2014||23 Jun 2015||Abbvie Inc.||Dual specific binding proteins directed against IL-1 and/or IL-17|
|US9067996||15 Mar 2013||30 Jun 2015||Abbvie Inc.||Antibodies to receptor for advanced glycation end products (RAGE) and uses thereof|
|US9096877||7 Oct 2010||4 Aug 2015||Macrogenics, Inc.||Fc region-containing polypeptides that exhibit improved effector function due to alterations of the extent of fucosylation, and methods for their use|
|US9102722||23 Sep 2013||11 Aug 2015||AbbVie Deutschland GmbH & Co. KG||Composition and method for the diagnosis and treatment of diseases associated with neurite degeneration|
|US9109026||3 Jun 2009||18 Aug 2015||Abbvie, Inc.||Dual variable domain immunoglobulins and uses thereof|
|US9115195||1 Mar 2011||25 Aug 2015||Abbvie Inc.||Therapeutic DLL4 binding proteins|
|US9120862||26 Jul 2010||1 Sep 2015||Abbott Laboratories||Antibodies relating to PIVKA-II and uses thereof|
|US9120870||28 Dec 2012||1 Sep 2015||Abbvie Inc.||Dual specific binding proteins directed against IL-13 and IL-17|
|US9132190||26 Nov 2013||15 Sep 2015||Abbvie Inc.||Therapeutic DLL4 binding proteins|
|US9163093||31 Oct 2013||20 Oct 2015||Abbvie Inc.||Anti-DLL4/VEGF dual variable domain immunoglobulin and uses thereof|
|US9175075||8 Dec 2010||3 Nov 2015||AbbVie Deutschland GmbH & Co. KG||Methods of treating retinal nerve fiber layer degeneration with monoclonal antibodies against a retinal guidance molecule (RGM) protein|
|US9176150||1 Aug 2011||3 Nov 2015||AbbVie Deutschland GmbH & Co. KG||Amyloid beta(1-42) oligomers, derivatives thereof and antibodies thereto, methods of preparation thereof and use thereof|
|US9272032||26 Nov 2014||1 Mar 2016||Medimmune, Llc||Stabilized liquid anti-RSV antibody formulations|
|US9283274||27 Sep 2013||15 Mar 2016||Medimmune Limited||RSV specific binding molecule|
|US9303085||12 Dec 2013||5 Apr 2016||Abbvie Inc.||IL-1 binding proteins|
|US9321831||13 Sep 2013||26 Apr 2016||Medimmune Limited||RSV-specific binding molecules and means for producing them|
|US9359430||15 Apr 2013||7 Jun 2016||Abbvie Inc.||Abeta conformer selective anti-Abeta globulomer monoclonal antibodies|
|US9365643||23 Dec 2014||14 Jun 2016||AbbVie Deutschland GmbH & Co. KG||Antibodies that bind to repulsive guidance molecule A (RGMA)|
|US9371374||23 Dec 2013||21 Jun 2016||Abbott Laboratories||HCV core lipid binding domain monoclonal antibodies|
|US9394360||14 Oct 2014||19 Jul 2016||Abbvie Inc.||Aβ conformer selective anti-Aβ globulomer monoclonal antibodies|
|US9394363||27 Nov 2012||19 Jul 2016||AbbVie Deutschland GmbH & Co. KG||Antibodies to receptor of advanced glycation end products (RAGE) and uses thereof|
|US9409986||18 Dec 2013||9 Aug 2016||Abbvie Inc.||IL-1 binding proteins|
|US9441038||17 Dec 2013||13 Sep 2016||Abbvie Inc.||IL-1 binding proteins|
|US9447183||16 Dec 2013||20 Sep 2016||Abbvie Inc.||IL-1 binding proteins|
|US9447184||26 Dec 2013||20 Sep 2016||Abbvie Inc.||IL-1 binding proteins|
|US9469686||15 Mar 2013||18 Oct 2016||Abbott Laboratories||Anti-GP73 monoclonal antibodies and methods of obtaining the same|
|US9469688||10 Aug 2015||18 Oct 2016||Abbvie Inc.||Therapeutic DLL4 binding proteins|
|US9469689||14 Jul 2015||18 Oct 2016||Abbvie Inc.||Therapeutic DLL4 binding proteins|
|US9481735||24 Apr 2014||1 Nov 2016||Abbvie Inc.||IL-17 binding proteins|
|US9481736||28 Apr 2014||1 Nov 2016||Abbvie, Inc.||IL-17 binding proteins|
|US9493560||19 Dec 2013||15 Nov 2016||Abbvie Inc.||Dual variable domain immunoglobulins and uses thereof|
|US9505829||19 Aug 2011||29 Nov 2016||Zoetis Belgium S.A.||Anti-NGF antibodies and their use|
|US9540432||14 May 2013||10 Jan 2017||AbbVie Deutschland GmbH & Co. KG||Anti-Aβ globulomer 7C6 antibodies|
|US9592293||9 Dec 2013||14 Mar 2017||Abbvie Inc.||Interleukin-13 binding proteins|
|US9605069||7 Jan 2015||28 Mar 2017||AbbVie Deutschland GmbH & Co. KG||Antibodies against the RGM a protein and uses thereof|
|US9617334||6 Jun 2013||11 Apr 2017||Zoetis Services Llc||Caninized anti-NGF antibodies and methods thereof|
|US9636398||13 Dec 2012||2 May 2017||AbbVie Deutschland GmbH & Co. KG||Composition and method for the diagnosis and treatment of iron-related disorders|
|US9663587||25 Apr 2014||30 May 2017||Abbvie Inc.||IL-17 binding proteins|
|US9670276||12 Jul 2013||6 Jun 2017||Abbvie Inc.||IL-1 binding proteins|
|US9790478||23 Dec 2013||17 Oct 2017||Abbott Laboratories||HCV NS3 recombinant antigens and mutants thereof for improved antibody detection|
|US20090220646 *||12 Jan 2007||3 Sep 2009||Ab Emzymes Gmbh||Enzyme Formulations For Animal Feed|
|US20140322330 *||9 Dec 2011||30 Oct 2014||Ns Technologies Pty Ltd||Methods for forming miniemulsions and use thereof for delivering bioactive agents|
|USRE45763||24 Sep 2013||20 Oct 2015||Abbott Laboratories||Antibodies to troponin I and methods of use thereof|
|International Classification||A61K9/22, A61K9/00, A61K9/20, A61K9/16|
|Cooperative Classification||A61K9/2013, A61K9/0024, A61K9/1617|
|European Classification||A61K9/20H4, A61K9/16H4, A61K9/00M5D|
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