CA2049342A1 - Formulations for stabilizing of igm antibodies - Google Patents
Formulations for stabilizing of igm antibodiesInfo
- Publication number
- CA2049342A1 CA2049342A1 CA 2049342 CA2049342A CA2049342A1 CA 2049342 A1 CA2049342 A1 CA 2049342A1 CA 2049342 CA2049342 CA 2049342 CA 2049342 A CA2049342 A CA 2049342A CA 2049342 A1 CA2049342 A1 CA 2049342A1
- Authority
- CA
- Canada
- Prior art keywords
- composition
- igm
- sodium
- lyophilized
- igm antibodies
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Abandoned
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/02—Inorganic compounds
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
- A61K39/39591—Stabilisation, fragmentation
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/16—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing nitrogen, e.g. nitro-, nitroso-, azo-compounds, nitriles, cyanates
- A61K47/18—Amines; Amides; Ureas; Quaternary ammonium compounds; Amino acids; Oligopeptides having up to five amino acids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/42—Proteins; Polypeptides; Degradation products thereof; Derivatives thereof, e.g. albumin, gelatin or zein
-
- 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
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
Abstract
Compositions suitable for intraveneous injection are disclosed, which comprise a stabilizing buffer solution containing IgM
antibodies. The compositions are effective in stabilizing the antibodies in solution, inhibiting precipitation and the formation of particulates in the final product vial, while maintaining a high level of immunoreactivity. The compositions can be lyophilized to form dry, stable products which can be readily reconstituted to provide injectable, particle-free antibody solutions.
antibodies. The compositions are effective in stabilizing the antibodies in solution, inhibiting precipitation and the formation of particulates in the final product vial, while maintaining a high level of immunoreactivity. The compositions can be lyophilized to form dry, stable products which can be readily reconstituted to provide injectable, particle-free antibody solutions.
Description
W090/11091 ,~ T/USgOt~1383 FOR~ULATIONS FOR
________________ _________ __ Back~round of the Invention ____ ______________________ It is well known that many protein preparations 05 intended for administration to humans require stabil-izers to prevent denaturation of the proteins, agglomeration and other alterations to the proteins prior to the use of the preparation. Many protein preparations are particularly unstable in dilute lO solutions. This instability is manifested in the formation of insoluble particles, and is often increased when the protein preparation is stored, or shipped.
A major challen-ge -that exists in the field of protein drugs is in the development of formulations that 15 maintain both protein solubility and activity.
Immunoglobulins, in particular, are recognized as possessing characteristics that tend to form partic-ulates in solution, requiring filtration of these formulations prior to using them for intra~enous in-20 jection. The formation of protein aggregates andparticulates has long been a problem in the development of parenteral immunoglobulin products. The admini-stration of immunoglobulin G (IgG~, for example, was limited to the intramuscular route be~ause of endogenous 25 anticomplementary activity due to aggregated immuno-globulin until the recent development of chemically and enzymatically treated immunoglobulin G. J. E.
Pennington, ~ev. Inf. Dis.,8(4):5371-5373 (1986).
Recent ~odifications in immunoglobulin G formulations 30 have helped to alleviate the problem. J. P. McCue et WO~0/11091 ~ ? PCT/US90/01383 6~ ~ , ,.
~.
al., Rev. Inf. Dis~, 8(4):5374-5381 (1986). However, most commercially available formulations now in use require iltration of the product prior to iniection to remove these insoluble aggregates or particulates.
05 The immuno~lobulin (IgM) isotype is, the largest of the immunoglobulins, having a ~olecular wei~ht of approximately 900,000 daltons. IgM mole!cules tend to be inherently unstable and precipitate reaclily upon being subjected to various forms of physical and chemical 10 stress. This characteristic makes the for~ulation of a stable composition containing IgM intended for parenteral administration di~ficult.
Summary of the Invention ______ _________________ The invention comprises stabilizing compositions 15 for IgM antibodies. The present compositions contain a buffer, human serum albumin, sodium chloride, and IgM
antibodies or antibody fragments. The compositions enhance the stability of IgM antibodies in solution intended for intravenous administration.
The present compositions can be lyophilized to form a dry powder. Lyophilization preserves the biological activity of the IgM antibody, and minimizes formation of particulates, which can occur in a liquid for~ulation under physical or chemical stress. The lyophilized 25 product can be readily reconstituted to a particle-free solution which shows no loss of biological activity, and which can be adminlstered without prior filtration.
The present liquid and lyophilized formulations both exhibit superior stabilizing characteristics in 30 terms of minimal protein particle formation, and . : : : , :
WO90/11091 P~T/US90/01383 ~' ^` :1 r~ ~. .~ rl preservation o~ immunoreactivity over time, and under stress conditions such as elevated temperatures, vial filling and shipping.
The present liquid and lyophilized compositions 05 have boeh been successful in stabilizing IgM antibodies.
The compositions maintain a pareicle-free, stable solution for injectable monoclonal antibodies and do not have to be filtered prior to administration. The lyophilized product, in particular, can be shipped and stored ~ithout loss of immunoreactivity. Neither formulation requires refrigeration or other special handling.
Brlef Descri~tion of the Fi~ures ___________________________ ____ Figure l shows gel filtration HPLC results 15 comparing non-lyophilized and lyophilized/reconstituted IgM formulations and placebos.
Figure 2 is a diagram of the results of an immuno-reactivity assay comparing the ability of non-lyophilized and lyophilized/reconstituted IgM
20 formulations to bind to solid phase lipid A.
Detailed Descri~tion of The Invention _______________ ____________._________ The compositions of this invention minimize the ~ormation of protein aggregates and particulates in reagents containing immunoglobulin M (IgM) antibodies 25 and insure that the antibody in solution maintains its immunoreactLvity over time. The preparation comprises a sterile, pharmaceutically acceptable solution containing tromethamine or phosphate buffer, having a neutral or .
WO90/11091 ~r~:~, PCT/US90/01383 .,, - ' :
~4-basic p~ (e.g., 6.8 or above), sodium chloride, IgM
antibodies and human serum albu~in.
Buffers ha~e long been used to stabilize the p~ of antibody products for parenteral injection. Protein 05 solubility in the buffer solution depends upon a number of factors, such as ionic strength and pH of the solution.
Buffers which can be used for this formulation include tromethamine and phosphate buffers ha~ing a lO neutral or basic pH. Lower p~ formulations showed less stability, i.e., a higher tendency to form aggregates.
Tromethamine is described in the Merck Index, 10th edition, ~erck and Co., Inc., Rahway, N.J. The concen-tration of tromethamine can be from about 5 to about lO0 15 mM, ha~ing a pH from about 8 to about lO.
A phosphate buffer, such as sodium phosphate, can also be used. A concentration of from about 8:to about 20 mM phosphate can be used in the present composition, having a pH of rrom about 6.8 to about 7.4.
A stabilizing protein is added to the formulation.
Stabilizing proteins are proteins which increase the solubility and/or stability of immunoglobulins in aqueous solutions. For example, when added to an aqueous solution of im~unoglobulins, these proteins 25 prevent the ammunoglobulins from precipitating out of the solution, thereby permitting higher concentrations of immunoglobulins to be solubilized. It has been found for the present compositions that human serum albumin (HSA) is a particularly useful stabilizer for IgM for 30 both liquid and lyophilized formulations. HSA is present in the formulation in an amount of about 2.5 to '' . ' . ~ ' WO 90/11091 c~ r ,, P.C~ S90/01383 about 10~ by weight per volume. Levels of HSA of from about 2.5~ (w/v), to about 5~ (w/v), are particularly effective in maintaining a stable solution of IgM.
In one embodiment of the invention stabilizing 05 reagents for HSA, e.g., sodium caprylate and N-acetyl tryptophanate, are present in the formation. HSA is less stable in solution (i.e., more likely to aggregate) in the absence of these compounds. For example, a 25 solution (w/v) oi` HSA contains 20 mM ~odium caprylate 10 and 20 mM N-acetyl tryptophanate, therefore, 2.5~ (w/v) HSA added to a formulation includes 2 mM sodium capylate and 2 mM N-acetyl tryptophanate. Other stabilizing reagents can be used other than N-acetyl tryptophanate and sodium caprylate, which are mentioned above for 15 illustrative purposes.
Sodium chloride is added to the present composition to increase the ionic strength which is required for the solubility of the IgM proteins. IgM proteins are more soluble in an aqueous salt solution than in water alone.
20 The amount of sodium chloride added is from about 200 to about 350 mM. About 270-300 mM sodium chloride is particularly effective for this purpose.
The present liquid and lyo,ohilized compositions can be used to stabilize all subclasses of IgM antibodies, 25 as well as I~M. The present compositions are particularly useful in stabilizing human monoclonal IgM
antibodies.
One embodiment of this invention comprises a composi~ion containin~ from about 5mM to about 100mM
30 tromethamine (pH 8-10), from about 200 mM to about 300mM
sodiu~ chloride, from about 2.5 to about 5% (w/v) HSA
and from about 2.5 to about 10.Omg/ml IgM antibody.
WO90/1~091 ~ PCT/US90/01383 ~ .
Sodium caprylate in an amount of from about 2 mM eo about 4 mM, and N-acetyl tryptophanate in an amount of from about 2 mM eo about 4 mM can, optionally, be included to stabilize the HSA. A preferred embodiment 05 of the invention comprises about 4.5 mM tromethamiQe (pH
8.5) about 270 mM sodium chloride, about 2.5% (w/v) HSA, about 5 mg/ml IgM antibodies or antibody fragments, and about 2 mH each of N-aceeyl tryptophanate and sodium caprylate. This formulation enhances the stability of lO immunological activity of the monoclonal antibody, and prevents the immunoglobulins in solution intended for intravenous administration to human subjects from precipitating and forming particulates in the final product vial. ~
Another embodiment of the present invention com-prises a composition containing from about 8 mM to about 20 mM of sterile, pyrogen-free sodium phosphate.~pH
6.8-7.4), from about 250 mM to about 350 mM sodium ohloride, from about 2.5 to about 5~ (w/v) HSA and from 20 about 2.5 to about 10.0 mg/ml IgM antibody or antibody fragments. Sodium caprylate and N-acetyl tryptophanate may be included in the formulation in the amount of about 2 mM to about 4 ~M of each. A preferred embodiment of this formulation comprises about 8 mH
25 sodium phosphate (pH 7.2), about 270 mM sodium chloride, about 5.0~ ~w/v) human serum albumin, about 5 mg/ml IgM
an~ibodies or antibody fragments, and about 2 mM each of sodium caprylate and N-acetyl tryptophanate.
In another embodiment of the present invention, the 30 above formulations can be lyophilized to form a dry, storable powder, which can be easily reconstituted to a ...... :
, .
~ ?
particle free solution suitable for intravenous injection. Lyophilization is a freeze drying process which is often used in the preparation of pharmaceutical products to preserve their biological ac~:ivity The 05 liquid composition is prepared, then lyophilized to form a dry cake-like product. The process generally involves drying a previously frozen sample in a vacuum to remove the ice, leaving the non-water components intact, in the form of a powdery or cake-like substance. The 10 lyophilized product can be stored ~or prolonged periods of time, and at elevated temperatures, without loss of biological activity, and can be readily reconstituted into a particle-free solution by the addition of an appropriate diluent. An appropriate diluent can be any 15 liquid which is biologically acceptable and in which the lyophilized powder is completely soluble Water, particularly sterile, pyrogen-free water, is the preferred diluent, since it does not include salts or other compounds which may affect the stability of the 20 antibody. The advanta~e of lyophilization is that the water content is reduced to levels which greatly reduce the various molecular events which lead to instability of the product. The lyophilized product is also better able to withstand the physical stresses of shipping. The . 25 reconstituted product is particle free, so it can be ad~inistered intravenously without prior filtration.
The present invention is further illustrated by the follow~ng Examples, which is not intended to be limiting in any way.
, . :~
WO90/11091 ,~ ~CT/US90/01383 6~ . .
EXAMPL~ l _ _ _ _ _ _ __ _ Pre~aration of I~ _Li~_id__n__Lyophilized Formulations Based On Tromethamine ___________ _________ Lig~id Formulation 05 IgM (HA-IA IgM, lot ~ 012567, Centocor, Inc., Malvern, PA) was concentrated to 5.5 mgJml using a Centriprep 30 Concentrator (Amicon). The concentrated protein (20 ml) was dispensed into a 25 ml graduated cylinder and 2 ml of HSA containing sodium caprylate and l0 N-acetyl tryptophanate (U.S.P. 25~ HSA in 20 mM sodium caprylate and 20 mM N-acetyl tryptophanate, Armour Pharmaceutical Co.), tromethamine (50 mM, pH 8.50) and 300 mM sodium chloride (NaCl) were added. The solution was filtered with a 0.2~ syrin~e filter into a 50 ml lS centrifuge tube. Sodium azide ( 0.22 ml of l0~
solution) was added to a final concentration of 0.l~.
The final solution, was a clear, light y2110w liquid composed of 4.95 mg/ml IgM, 45 mM tromethamine (pH
; 8.35), 270 mM sodium chloride, 2.5~ HSA, 2 mM sodium 20 caprylate and 2 mM N-acetyl tryptophanate A "placebo" formulation was also made, which was exacely the same as the above formulation except that the I8M was left out.. Thus, the placebo formulation contained 45 mM Tris buffer (pH 8.35)~ 270 mM NaCl, 2.5 25 HSA, 2 mM sodium caprylate and 2 mM N-acetyl trypto-phanate.
Lvu~hilization The liq~id IgM formulation was dispensed in l ml increments into 2 ml Type l Tubing vials (~est Co.). A
30 total of 20 vials were filled. The vials were placed in , 1., i, .
, WO9~111091 PCT/US90/01383 ,~ ~
a lyophiliæer (FTS) havin~ a l' x l' shelf. In order to generate a full thermal load, the remainder of the shelf space was loaded with placebo vials.
The vials were capped with 13 mM gray butyl 05 lyophilization closures (#224142, Uheaton). The shelves of the lyophili7er were prechilled to about 5C, ~ 2C.
The test vials and placebo ~ials were loaded onto a shelf in a tray, and a sli~ht vaccuum was induced in the chamber to maintain a good door seal. A eotal of 20 lO vials of product and 355 vials of placebo were filled to occupy the entire shelf space.
After the vials had reached and maintained 5C for at least l hour, the shelf surface temperature was set for about -40~C.- The-vials were-a~owed-to-remain at - `
lS -40C for at least one hour. The condenser was chilled to about -70C. The pressure in the chamber was reduced by Means of a mechanical pump to less than 50 Torr. The shelf surface temperature was re~ulated such that the product temperature remained between -47C and -42C.
2Q After the product temperature had reached and maintained the shelf temperature for at least one hour, a mass spectrum of the residual gasses in the chamber was rscorded. The shelf surface temperature was then set for about ~20C. When the temperature reached and 25 maintained +20C for at least 2 hours, the partial pressures of the residual gasses in the chamber was recorded.
The chamber was then backfilled with dry nitrogen to a pressure of about 600 Torr. The product was 30 removed from the dryer and crimp seals were applied to . .
, -.: .
WOg~/11091 PCTtUS90/~1383 the vials. The formulated protein formed a very dense cake upon freezing.
The lyophilized cakes that were formed did not possess any crust or glaze on the surface, and were 05 uniform throughout the vial.
Reconstitution of Lvophilized I~M
The crimp seals were removed from t:he vials to expose the closure, and the closure was removed from one vial containing the product and one placebo vial. A
lO sterile pipette was filled with l.O ml of sterile/pyrogen-free (s/pf) water (McGaw), which was dispensed into the vial holding the lyophilized product.
Once all of the water was injected, the length of ti~e necessary to dissolve all visually observable material lS ~as measured:
Reconstitution Times ____________________ Placebo Product 0.5 minutes 0.3 minutes Visual Examination After_Reconstitution The vials were held directly in front of a black background for ~isual examination. This was accom-plished by placing a light source below the vial so that the beam o f light proceded upwards through the liquid.
Changes in color, turbidity, flocculation, fine precipi-25 tation or any other pareiculate matter were examined.
No discernable difference could be seen between the non-lyophilized formulation and the lyophilized ` . ' - ' , . ` ~
':
. .
: .
::
. ~ ,, ~ . .
WO90/11091 PCT/US90/0l383 ;J
J~ j reconstituted formulation. The results are shown in the following Table:
Visual Particle & Cosmetic Analvsis Vial Appearance 05 Place.bo (non-lyophilized) clear, yellowish liquid Product (non-lyophilized) clear, yellowish liquid Placebo (lyophilized) clear, yellowish liquid Product (lyophilized) clear, yellowish liquid HPLC Gel Filtration ___________________ The lypophilized product and placebo were measured by HPLC (Waters) gel filtration. Non-lyophilized product ~- --and placebo were also run. A DuPont Zorbax GF-450 gel column was equilibrated with a mixture of 0.2M sodium phosphate buffer (pH 6.8) and 0.3 M NaCl at a flow rate 15 of 1 ml/min. Absorbance wavelength was set for 214 nm.
One ~1 of undiluted sample (lyophilized and pre-lyophilized product and placebo) was injected onto the column through an automatic injector and run for 15 minutes. The results, shown in Figure 1, indicated no 20 detectable difference between the pre-lyophilized placebo and product and lyophilized/reconstitut~ed placebo and product.
Immunoactivity Assay _____ _______ _ ___ The immunological activity of the IgM in each 25 formulation was determined using an enzyme-linked immunoassay to measure binding to solid-phase lipid A.
': .. . .:
, .:
.
r~ ` :.. .
. -12-A vial of Salmonella minnesota R595 lipid A (List Biological Laboratories, Inc., Campbell, CA; catalog #401) was reconstituted to 1 mg/ml with 0.5~ TEA
~triethylamine) in s/pf water. A 10 ~g/ml solution of 05 lipid A was made in a buffer solution consisting o 10 mM HEPES and sterile/pyrogen-free 0.9~ NaCl ~s/pf saline, McGraw), pH 7.2 (Buffer #1). This formuation was then dispensed into a PVC microtiter plate (Dynatech Laboratories, Inc., Chantilly, VA catalog ~011-010- , 10 2101), 50 ~l/well, and the plate was covered and incubated overnight at 4C.
The plates were removed from the incubator and washed three times with s/pf saline, then blocked by .... ...d.ispensing 200.~1/well-of a buffer consisting of: 10 mM
15 HEPES, s/pf saline and 2~ heat-inactivated FBS, pH 7.2 (Buffer ~2). The plates were covered and incubaeed for 1 hour at 37C. After incubation, the plates were washed three times with s/pf saline.
Solutions were prepared of the test formulations, 20 an IgM standard and a negative control of human myeloma IgM (Chrompure Human Myeloma IgM, Jackson Immuno Research Laboratories, West Grove, PA) at a concentration of 5.0 ~g/ml.
Buffer #2 was dispensed into the wells in rows B-H
25 (50 ~l/well). The IgM standard was dispensed into row A, columns 1-3 (100 ~l/well). Test formulations were dispensed in triplicate in row A, columns 4-12 (100 ~l/well). Serial 50~1 dilutions were then made down the rows of the plate to row H. 50 ~1 of ehe 100 ~1 in row 30 H was discarded, and the negative control was added.
. .
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. . , , ~ .
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WO90/11~91 PCT/~'S90/01383 -l3~
The plate was covered and incubated for 2 hours at 37C, then washed three times with s/pf saline.
Substrate solution was prepared by ,adding l phosphatase substrate table tsigma Chemicals, Inc., St.
05 Louis, MO) to 5 ml of s/pf water containing a l:500 dilution of an alkaline buffer solution (Sigma Chemicals, catalog #014-105), and incubated for 20 minutes. The reaction was stopped by adding 50 ~l of 3M
NaOH.
The optical density of the solutions were measured at 414 nm or using a plate reader. The data were analyzed using a 4 parameter fit of OD versus concentration.
The results, shown in Figure 2, indicated no detectable difference between the activity of the prelyophilized and lyophilized product.
Tem~erature Stress Testin~
Lyophilized product samples were stored at 4~, 22~C and 40C. The samples were evaluated periodically for activity and appearance (i.e., particulate formation). The samples were reconstituted prior to evaluation. The results are shown in the following table:
Tem~erature/Time Activity Ap~earance 25 4C/2 months no change clear 22-C/2 months no change clear 40C/2 months slight decrease some particulates4C/5 months no change clear . ::
: ........................ : , ~ : . . . . .. .
- . : :. ~ ., :
: . . . , : :
:
. .
WO90/11091 PCT~US90/01383 ~ ' ,,~
22C/5 months no change clear 4C/7 months no change clezr E~uivalents Those skilled in the art will recognize, or be able 05 to ascertain, using no more than routine experiment-ation, many equivalents co the specific embodiments of the invention described herein. These and all other equivalents are intended to be encompassed by the following claims.
:. ' ~: : :
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________________ _________ __ Back~round of the Invention ____ ______________________ It is well known that many protein preparations 05 intended for administration to humans require stabil-izers to prevent denaturation of the proteins, agglomeration and other alterations to the proteins prior to the use of the preparation. Many protein preparations are particularly unstable in dilute lO solutions. This instability is manifested in the formation of insoluble particles, and is often increased when the protein preparation is stored, or shipped.
A major challen-ge -that exists in the field of protein drugs is in the development of formulations that 15 maintain both protein solubility and activity.
Immunoglobulins, in particular, are recognized as possessing characteristics that tend to form partic-ulates in solution, requiring filtration of these formulations prior to using them for intra~enous in-20 jection. The formation of protein aggregates andparticulates has long been a problem in the development of parenteral immunoglobulin products. The admini-stration of immunoglobulin G (IgG~, for example, was limited to the intramuscular route be~ause of endogenous 25 anticomplementary activity due to aggregated immuno-globulin until the recent development of chemically and enzymatically treated immunoglobulin G. J. E.
Pennington, ~ev. Inf. Dis.,8(4):5371-5373 (1986).
Recent ~odifications in immunoglobulin G formulations 30 have helped to alleviate the problem. J. P. McCue et WO~0/11091 ~ ? PCT/US90/01383 6~ ~ , ,.
~.
al., Rev. Inf. Dis~, 8(4):5374-5381 (1986). However, most commercially available formulations now in use require iltration of the product prior to iniection to remove these insoluble aggregates or particulates.
05 The immuno~lobulin (IgM) isotype is, the largest of the immunoglobulins, having a ~olecular wei~ht of approximately 900,000 daltons. IgM mole!cules tend to be inherently unstable and precipitate reaclily upon being subjected to various forms of physical and chemical 10 stress. This characteristic makes the for~ulation of a stable composition containing IgM intended for parenteral administration di~ficult.
Summary of the Invention ______ _________________ The invention comprises stabilizing compositions 15 for IgM antibodies. The present compositions contain a buffer, human serum albumin, sodium chloride, and IgM
antibodies or antibody fragments. The compositions enhance the stability of IgM antibodies in solution intended for intravenous administration.
The present compositions can be lyophilized to form a dry powder. Lyophilization preserves the biological activity of the IgM antibody, and minimizes formation of particulates, which can occur in a liquid for~ulation under physical or chemical stress. The lyophilized 25 product can be readily reconstituted to a particle-free solution which shows no loss of biological activity, and which can be adminlstered without prior filtration.
The present liquid and lyophilized formulations both exhibit superior stabilizing characteristics in 30 terms of minimal protein particle formation, and . : : : , :
WO90/11091 P~T/US90/01383 ~' ^` :1 r~ ~. .~ rl preservation o~ immunoreactivity over time, and under stress conditions such as elevated temperatures, vial filling and shipping.
The present liquid and lyophilized compositions 05 have boeh been successful in stabilizing IgM antibodies.
The compositions maintain a pareicle-free, stable solution for injectable monoclonal antibodies and do not have to be filtered prior to administration. The lyophilized product, in particular, can be shipped and stored ~ithout loss of immunoreactivity. Neither formulation requires refrigeration or other special handling.
Brlef Descri~tion of the Fi~ures ___________________________ ____ Figure l shows gel filtration HPLC results 15 comparing non-lyophilized and lyophilized/reconstituted IgM formulations and placebos.
Figure 2 is a diagram of the results of an immuno-reactivity assay comparing the ability of non-lyophilized and lyophilized/reconstituted IgM
20 formulations to bind to solid phase lipid A.
Detailed Descri~tion of The Invention _______________ ____________._________ The compositions of this invention minimize the ~ormation of protein aggregates and particulates in reagents containing immunoglobulin M (IgM) antibodies 25 and insure that the antibody in solution maintains its immunoreactLvity over time. The preparation comprises a sterile, pharmaceutically acceptable solution containing tromethamine or phosphate buffer, having a neutral or .
WO90/11091 ~r~:~, PCT/US90/01383 .,, - ' :
~4-basic p~ (e.g., 6.8 or above), sodium chloride, IgM
antibodies and human serum albu~in.
Buffers ha~e long been used to stabilize the p~ of antibody products for parenteral injection. Protein 05 solubility in the buffer solution depends upon a number of factors, such as ionic strength and pH of the solution.
Buffers which can be used for this formulation include tromethamine and phosphate buffers ha~ing a lO neutral or basic pH. Lower p~ formulations showed less stability, i.e., a higher tendency to form aggregates.
Tromethamine is described in the Merck Index, 10th edition, ~erck and Co., Inc., Rahway, N.J. The concen-tration of tromethamine can be from about 5 to about lO0 15 mM, ha~ing a pH from about 8 to about lO.
A phosphate buffer, such as sodium phosphate, can also be used. A concentration of from about 8:to about 20 mM phosphate can be used in the present composition, having a pH of rrom about 6.8 to about 7.4.
A stabilizing protein is added to the formulation.
Stabilizing proteins are proteins which increase the solubility and/or stability of immunoglobulins in aqueous solutions. For example, when added to an aqueous solution of im~unoglobulins, these proteins 25 prevent the ammunoglobulins from precipitating out of the solution, thereby permitting higher concentrations of immunoglobulins to be solubilized. It has been found for the present compositions that human serum albumin (HSA) is a particularly useful stabilizer for IgM for 30 both liquid and lyophilized formulations. HSA is present in the formulation in an amount of about 2.5 to '' . ' . ~ ' WO 90/11091 c~ r ,, P.C~ S90/01383 about 10~ by weight per volume. Levels of HSA of from about 2.5~ (w/v), to about 5~ (w/v), are particularly effective in maintaining a stable solution of IgM.
In one embodiment of the invention stabilizing 05 reagents for HSA, e.g., sodium caprylate and N-acetyl tryptophanate, are present in the formation. HSA is less stable in solution (i.e., more likely to aggregate) in the absence of these compounds. For example, a 25 solution (w/v) oi` HSA contains 20 mM ~odium caprylate 10 and 20 mM N-acetyl tryptophanate, therefore, 2.5~ (w/v) HSA added to a formulation includes 2 mM sodium capylate and 2 mM N-acetyl tryptophanate. Other stabilizing reagents can be used other than N-acetyl tryptophanate and sodium caprylate, which are mentioned above for 15 illustrative purposes.
Sodium chloride is added to the present composition to increase the ionic strength which is required for the solubility of the IgM proteins. IgM proteins are more soluble in an aqueous salt solution than in water alone.
20 The amount of sodium chloride added is from about 200 to about 350 mM. About 270-300 mM sodium chloride is particularly effective for this purpose.
The present liquid and lyo,ohilized compositions can be used to stabilize all subclasses of IgM antibodies, 25 as well as I~M. The present compositions are particularly useful in stabilizing human monoclonal IgM
antibodies.
One embodiment of this invention comprises a composi~ion containin~ from about 5mM to about 100mM
30 tromethamine (pH 8-10), from about 200 mM to about 300mM
sodiu~ chloride, from about 2.5 to about 5% (w/v) HSA
and from about 2.5 to about 10.Omg/ml IgM antibody.
WO90/1~091 ~ PCT/US90/01383 ~ .
Sodium caprylate in an amount of from about 2 mM eo about 4 mM, and N-acetyl tryptophanate in an amount of from about 2 mM eo about 4 mM can, optionally, be included to stabilize the HSA. A preferred embodiment 05 of the invention comprises about 4.5 mM tromethamiQe (pH
8.5) about 270 mM sodium chloride, about 2.5% (w/v) HSA, about 5 mg/ml IgM antibodies or antibody fragments, and about 2 mH each of N-aceeyl tryptophanate and sodium caprylate. This formulation enhances the stability of lO immunological activity of the monoclonal antibody, and prevents the immunoglobulins in solution intended for intravenous administration to human subjects from precipitating and forming particulates in the final product vial. ~
Another embodiment of the present invention com-prises a composition containing from about 8 mM to about 20 mM of sterile, pyrogen-free sodium phosphate.~pH
6.8-7.4), from about 250 mM to about 350 mM sodium ohloride, from about 2.5 to about 5~ (w/v) HSA and from 20 about 2.5 to about 10.0 mg/ml IgM antibody or antibody fragments. Sodium caprylate and N-acetyl tryptophanate may be included in the formulation in the amount of about 2 mM to about 4 ~M of each. A preferred embodiment of this formulation comprises about 8 mH
25 sodium phosphate (pH 7.2), about 270 mM sodium chloride, about 5.0~ ~w/v) human serum albumin, about 5 mg/ml IgM
an~ibodies or antibody fragments, and about 2 mM each of sodium caprylate and N-acetyl tryptophanate.
In another embodiment of the present invention, the 30 above formulations can be lyophilized to form a dry, storable powder, which can be easily reconstituted to a ...... :
, .
~ ?
particle free solution suitable for intravenous injection. Lyophilization is a freeze drying process which is often used in the preparation of pharmaceutical products to preserve their biological ac~:ivity The 05 liquid composition is prepared, then lyophilized to form a dry cake-like product. The process generally involves drying a previously frozen sample in a vacuum to remove the ice, leaving the non-water components intact, in the form of a powdery or cake-like substance. The 10 lyophilized product can be stored ~or prolonged periods of time, and at elevated temperatures, without loss of biological activity, and can be readily reconstituted into a particle-free solution by the addition of an appropriate diluent. An appropriate diluent can be any 15 liquid which is biologically acceptable and in which the lyophilized powder is completely soluble Water, particularly sterile, pyrogen-free water, is the preferred diluent, since it does not include salts or other compounds which may affect the stability of the 20 antibody. The advanta~e of lyophilization is that the water content is reduced to levels which greatly reduce the various molecular events which lead to instability of the product. The lyophilized product is also better able to withstand the physical stresses of shipping. The . 25 reconstituted product is particle free, so it can be ad~inistered intravenously without prior filtration.
The present invention is further illustrated by the follow~ng Examples, which is not intended to be limiting in any way.
, . :~
WO90/11091 ,~ ~CT/US90/01383 6~ . .
EXAMPL~ l _ _ _ _ _ _ __ _ Pre~aration of I~ _Li~_id__n__Lyophilized Formulations Based On Tromethamine ___________ _________ Lig~id Formulation 05 IgM (HA-IA IgM, lot ~ 012567, Centocor, Inc., Malvern, PA) was concentrated to 5.5 mgJml using a Centriprep 30 Concentrator (Amicon). The concentrated protein (20 ml) was dispensed into a 25 ml graduated cylinder and 2 ml of HSA containing sodium caprylate and l0 N-acetyl tryptophanate (U.S.P. 25~ HSA in 20 mM sodium caprylate and 20 mM N-acetyl tryptophanate, Armour Pharmaceutical Co.), tromethamine (50 mM, pH 8.50) and 300 mM sodium chloride (NaCl) were added. The solution was filtered with a 0.2~ syrin~e filter into a 50 ml lS centrifuge tube. Sodium azide ( 0.22 ml of l0~
solution) was added to a final concentration of 0.l~.
The final solution, was a clear, light y2110w liquid composed of 4.95 mg/ml IgM, 45 mM tromethamine (pH
; 8.35), 270 mM sodium chloride, 2.5~ HSA, 2 mM sodium 20 caprylate and 2 mM N-acetyl tryptophanate A "placebo" formulation was also made, which was exacely the same as the above formulation except that the I8M was left out.. Thus, the placebo formulation contained 45 mM Tris buffer (pH 8.35)~ 270 mM NaCl, 2.5 25 HSA, 2 mM sodium caprylate and 2 mM N-acetyl trypto-phanate.
Lvu~hilization The liq~id IgM formulation was dispensed in l ml increments into 2 ml Type l Tubing vials (~est Co.). A
30 total of 20 vials were filled. The vials were placed in , 1., i, .
, WO9~111091 PCT/US90/01383 ,~ ~
a lyophiliæer (FTS) havin~ a l' x l' shelf. In order to generate a full thermal load, the remainder of the shelf space was loaded with placebo vials.
The vials were capped with 13 mM gray butyl 05 lyophilization closures (#224142, Uheaton). The shelves of the lyophili7er were prechilled to about 5C, ~ 2C.
The test vials and placebo ~ials were loaded onto a shelf in a tray, and a sli~ht vaccuum was induced in the chamber to maintain a good door seal. A eotal of 20 lO vials of product and 355 vials of placebo were filled to occupy the entire shelf space.
After the vials had reached and maintained 5C for at least l hour, the shelf surface temperature was set for about -40~C.- The-vials were-a~owed-to-remain at - `
lS -40C for at least one hour. The condenser was chilled to about -70C. The pressure in the chamber was reduced by Means of a mechanical pump to less than 50 Torr. The shelf surface temperature was re~ulated such that the product temperature remained between -47C and -42C.
2Q After the product temperature had reached and maintained the shelf temperature for at least one hour, a mass spectrum of the residual gasses in the chamber was rscorded. The shelf surface temperature was then set for about ~20C. When the temperature reached and 25 maintained +20C for at least 2 hours, the partial pressures of the residual gasses in the chamber was recorded.
The chamber was then backfilled with dry nitrogen to a pressure of about 600 Torr. The product was 30 removed from the dryer and crimp seals were applied to . .
, -.: .
WOg~/11091 PCTtUS90/~1383 the vials. The formulated protein formed a very dense cake upon freezing.
The lyophilized cakes that were formed did not possess any crust or glaze on the surface, and were 05 uniform throughout the vial.
Reconstitution of Lvophilized I~M
The crimp seals were removed from t:he vials to expose the closure, and the closure was removed from one vial containing the product and one placebo vial. A
lO sterile pipette was filled with l.O ml of sterile/pyrogen-free (s/pf) water (McGaw), which was dispensed into the vial holding the lyophilized product.
Once all of the water was injected, the length of ti~e necessary to dissolve all visually observable material lS ~as measured:
Reconstitution Times ____________________ Placebo Product 0.5 minutes 0.3 minutes Visual Examination After_Reconstitution The vials were held directly in front of a black background for ~isual examination. This was accom-plished by placing a light source below the vial so that the beam o f light proceded upwards through the liquid.
Changes in color, turbidity, flocculation, fine precipi-25 tation or any other pareiculate matter were examined.
No discernable difference could be seen between the non-lyophilized formulation and the lyophilized ` . ' - ' , . ` ~
':
. .
: .
::
. ~ ,, ~ . .
WO90/11091 PCT/US90/0l383 ;J
J~ j reconstituted formulation. The results are shown in the following Table:
Visual Particle & Cosmetic Analvsis Vial Appearance 05 Place.bo (non-lyophilized) clear, yellowish liquid Product (non-lyophilized) clear, yellowish liquid Placebo (lyophilized) clear, yellowish liquid Product (lyophilized) clear, yellowish liquid HPLC Gel Filtration ___________________ The lypophilized product and placebo were measured by HPLC (Waters) gel filtration. Non-lyophilized product ~- --and placebo were also run. A DuPont Zorbax GF-450 gel column was equilibrated with a mixture of 0.2M sodium phosphate buffer (pH 6.8) and 0.3 M NaCl at a flow rate 15 of 1 ml/min. Absorbance wavelength was set for 214 nm.
One ~1 of undiluted sample (lyophilized and pre-lyophilized product and placebo) was injected onto the column through an automatic injector and run for 15 minutes. The results, shown in Figure 1, indicated no 20 detectable difference between the pre-lyophilized placebo and product and lyophilized/reconstitut~ed placebo and product.
Immunoactivity Assay _____ _______ _ ___ The immunological activity of the IgM in each 25 formulation was determined using an enzyme-linked immunoassay to measure binding to solid-phase lipid A.
': .. . .:
, .:
.
r~ ` :.. .
. -12-A vial of Salmonella minnesota R595 lipid A (List Biological Laboratories, Inc., Campbell, CA; catalog #401) was reconstituted to 1 mg/ml with 0.5~ TEA
~triethylamine) in s/pf water. A 10 ~g/ml solution of 05 lipid A was made in a buffer solution consisting o 10 mM HEPES and sterile/pyrogen-free 0.9~ NaCl ~s/pf saline, McGraw), pH 7.2 (Buffer #1). This formuation was then dispensed into a PVC microtiter plate (Dynatech Laboratories, Inc., Chantilly, VA catalog ~011-010- , 10 2101), 50 ~l/well, and the plate was covered and incubated overnight at 4C.
The plates were removed from the incubator and washed three times with s/pf saline, then blocked by .... ...d.ispensing 200.~1/well-of a buffer consisting of: 10 mM
15 HEPES, s/pf saline and 2~ heat-inactivated FBS, pH 7.2 (Buffer ~2). The plates were covered and incubaeed for 1 hour at 37C. After incubation, the plates were washed three times with s/pf saline.
Solutions were prepared of the test formulations, 20 an IgM standard and a negative control of human myeloma IgM (Chrompure Human Myeloma IgM, Jackson Immuno Research Laboratories, West Grove, PA) at a concentration of 5.0 ~g/ml.
Buffer #2 was dispensed into the wells in rows B-H
25 (50 ~l/well). The IgM standard was dispensed into row A, columns 1-3 (100 ~l/well). Test formulations were dispensed in triplicate in row A, columns 4-12 (100 ~l/well). Serial 50~1 dilutions were then made down the rows of the plate to row H. 50 ~1 of ehe 100 ~1 in row 30 H was discarded, and the negative control was added.
. .
",~:, . , ' . . ' ' ' .
. . , , ~ .
:. ~ : :., :
WO90/11~91 PCT/~'S90/01383 -l3~
The plate was covered and incubated for 2 hours at 37C, then washed three times with s/pf saline.
Substrate solution was prepared by ,adding l phosphatase substrate table tsigma Chemicals, Inc., St.
05 Louis, MO) to 5 ml of s/pf water containing a l:500 dilution of an alkaline buffer solution (Sigma Chemicals, catalog #014-105), and incubated for 20 minutes. The reaction was stopped by adding 50 ~l of 3M
NaOH.
The optical density of the solutions were measured at 414 nm or using a plate reader. The data were analyzed using a 4 parameter fit of OD versus concentration.
The results, shown in Figure 2, indicated no detectable difference between the activity of the prelyophilized and lyophilized product.
Tem~erature Stress Testin~
Lyophilized product samples were stored at 4~, 22~C and 40C. The samples were evaluated periodically for activity and appearance (i.e., particulate formation). The samples were reconstituted prior to evaluation. The results are shown in the following table:
Tem~erature/Time Activity Ap~earance 25 4C/2 months no change clear 22-C/2 months no change clear 40C/2 months slight decrease some particulates4C/5 months no change clear . ::
: ........................ : , ~ : . . . . .. .
- . : :. ~ ., :
: . . . , : :
:
. .
WO90/11091 PCT~US90/01383 ~ ' ,,~
22C/5 months no change clear 4C/7 months no change clezr E~uivalents Those skilled in the art will recognize, or be able 05 to ascertain, using no more than routine experiment-ation, many equivalents co the specific embodiments of the invention described herein. These and all other equivalents are intended to be encompassed by the following claims.
:. ' ~: : :
,: . ~: ' ' ' ': ' : ' ' ' ' ' .-:, ~
Claims (29)
- l. A composition for IgM antibodies comprising a buffer, human serum albumin, sodium chloride, and IgM antibodies.
- 2. A composition of Claim 1, wherein the buffer com-prises phosphate buffer or tromethamine.
- 3. A composition of Claim 2, wherein the phosphate buffer comprises sodium phosphate having a concen-tration from about 8 mM and about 20 mM and having a pH from about 6.8 to about 7.4.
- 4. A composition of Claim 2, wherein the tromethamine buffer has a concentration from about 5 to about 100 mM and a pH from about 8 to about 10.
- 5. A composition of Claim 1, wherein the concentration of sodium chloride is about 270 mM.
- 6. A composition of Claim 1 further comprising N-acetyl tryptophanate and sodium caprylate.
- 7. A composition of Claim 7, wherein the concentration of N-acetyl tryptophanate is from about 2 mM to about 4 mM and the concentration of sodium caprylate is from about 2 mM to about 4 mM.
- 8. A composition of Claim 1, wherein the concentration of IgM antibodies is about 5.0 mg/ml.
- 9. A composition of Claim 8, wherein the IgM
antibodies are human immunoglobulin. - 10. A composition of Claim 1, wherein the IgM
antibodies are monoclonal antibodies. - 11. A composition of Claim 1 containing from about 2.5%
to about 5% (w/v) human serum albumin. - 12. A composition of Claim 1 which is lyophilized.
- 13. A composition of Claim 12 comprising a dry powder which can be reconstituted to yield an injectable solution of IgM.
- 14. An injectable composition for IgM, which comprises:
a. about 5 mM to about 100 mM tromethamine having a pH of from about 8 to about 10;
b. about 200 to about 300 mM sodium chloride;
c. about 2.5 to about 5% weight per volume human serum albumin; and d. about 2.5 to about 10.0 mg/ml of IgM
antibodies. - 15. An injectable composition of Claim 14, further comprising about 2 mM to about 4 mM sodium caprylate and about 2 mM to about 4 mM N-acetyl tryptophanate.
- 16. A composition of Claim 15 which comprises:
a. 45 mM tromethamine, pH 8.5;
b. 270 mM sodium chloride;
c. 2.5% weight per volume human serum albumin;
d. 5 mg/ml IgM antibodies;
e. 2 mM sodium caprylate; and f. 2 mM N-acetyl tryptophanate. - 17. A composition of Claim 16 which is lyophilized.
- 18. A composition of Claim 17 comprising a dry powder which can be reconstituted to yield an injectable solution of IgM.
- 19. An injectable composition of IgM, which comprises:
a. about 8 mM to about 20 mM sodium phosphate, having a pH of from about 6.8 to about 7.4;
b. about 250 to about 350 mM sodium chloride;
c. about 2.5 to about 5.0% weight per volume human serum albumin; and d. about 2.5 to about 10.0 mg/ml of IgM
antibodies. - 20. An injectable composition of Claim 19, further comprising about 2 mM to about 4 mM sodium caprylate and about 2 mM to about 4 mM N-acetyl tryptophanate.
- 21. A composition of Claim 20 which comprises:
a. 8 mM sodium phosphate, pH 7.2;
b. 270 mM sodium chloride;
c. 5.0% weight per volume human serum albumin;
d. 5 mg/ml IgM antibodies;
e. 2 mM sodium caprylate; and f. 2 mM N-acetyl tryptophanate. - 22. A composition of Claim 21 which is lyophilized.
- 23. A composition of Claim 22 comprising a dry powder which can be reconstituted to yield an injectable solution of IgM.
- 24. In an IgM composition containing buffer, protein and IgM, the improvement comprising combining the IgM antibodies with phosphate or tromethamine buffer, sodium chloride, and human serum albumin.
- 25. An improved composition of Claim 24, wherein the human serum albumin is stabilized with N-acetyl tryptophanate and sodium caprylate.
- 26. An improved composition of Claim 24, comprising:
a. about 5 mM to about 100 mM tromethamine having a pH of from about 8 to about 10;
b. about 200 to about 300 mM sodium chloride;
c. about 2.5 to about 5% (w/v) human serum albumin;
d. about 2.5 to about 10.0 mg/ml of IgM
antibodies.
e. about 2 mM to about 4 mM sodium caprylate; and f. about 2 mM to about 4 mM N-acetyl tryptophanate. - 27. An improved composition of Claim 24 comprising:
a. about 8 mM to about 20 mM sodium phosphate;
b. about 250 to about 350 mM sodium chloride;
c. about 2.5 to about 5.0% (w/v) human serum albumin;
d. about 2.5 to about 10.0 mg/ml of IgM
antibodies;
e. about 2 mM to about 4 mM sodium caprylate; and f. about 2 mM to about 4 mM N-acetyl tryptophanate. - 28. An improved composition of Claim 26 which is lyophilized to a dry powder, which powder can be reconstituted to yield an injectable solution of IgM.
- 29. An improved composition of Claim 27 which is lyophilized to a dry powder, which powder can be reconstituted to form an injectable solution of IgM.
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US32857989A | 1989-03-27 | 1989-03-27 | |
US328,579 | 1989-03-27 |
Publications (1)
Publication Number | Publication Date |
---|---|
CA2049342A1 true CA2049342A1 (en) | 1990-09-28 |
Family
ID=23281559
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CA 2049342 Abandoned CA2049342A1 (en) | 1989-03-27 | 1990-03-13 | Formulations for stabilizing of igm antibodies |
Country Status (4)
Country | Link |
---|---|
EP (1) | EP0465513A1 (en) |
JP (1) | JPH04504253A (en) |
CA (1) | CA2049342A1 (en) |
WO (1) | WO1990011091A1 (en) |
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EP0537222A1 (en) * | 1990-07-03 | 1993-04-21 | Akzo N.V. | Immunoreactive compound |
JPH0565233A (en) * | 1991-03-08 | 1993-03-19 | Mitsui Toatsu Chem Inc | Monoclonal antibody-containing lyophilized preparation |
CA2097932A1 (en) * | 1991-10-11 | 1993-04-12 | Shojiro Yamazaki | Antibody containing composition |
ATE146359T1 (en) * | 1992-01-21 | 1997-01-15 | Stanford Res Inst Int | IMPROVED METHOD FOR PRODUCING MICRONIZED POLYPEPTIDE DRUGS |
US6267958B1 (en) | 1995-07-27 | 2001-07-31 | Genentech, Inc. | Protein formulation |
US6171586B1 (en) | 1997-06-13 | 2001-01-09 | Genentech, Inc. | Antibody formulation |
US6991790B1 (en) | 1997-06-13 | 2006-01-31 | Genentech, Inc. | Antibody formulation |
WO1999037329A1 (en) * | 1998-01-22 | 1999-07-29 | Astrazeneca Ab | Pharmaceutical formulation comprising an antibody and a citrate buffer |
AU1344102A (en) | 2000-10-12 | 2002-04-22 | Genentech Inc | Reduced-viscosity concentrated protein formulations |
US8703126B2 (en) | 2000-10-12 | 2014-04-22 | Genentech, Inc. | Reduced-viscosity concentrated protein formulations |
US7682608B2 (en) | 2001-08-29 | 2010-03-23 | Chugai Seiyaku Kabushiki Kaisha | Stabilized preparations containing antibody |
PT1441589E (en) | 2001-11-08 | 2012-08-13 | Abbott Biotherapeutics Corp | Stable liquid pharmaceutical formulation of igg antibodies |
US20040033228A1 (en) | 2002-08-16 | 2004-02-19 | Hans-Juergen Krause | Formulation of human antibodies for treating TNF-alpha associated disorders |
BRPI0403964B8 (en) | 2003-04-04 | 2021-05-25 | Genentech Inc | stable liquid formulations, article of manufacture and use of these formulations for the treatment of ige-mediated dysfunction |
JP5322405B2 (en) * | 2007-06-07 | 2013-10-23 | 小林製薬株式会社 | Protein-containing composition |
US8883146B2 (en) | 2007-11-30 | 2014-11-11 | Abbvie Inc. | Protein formulations and methods of making same |
EP2403874A1 (en) | 2009-03-06 | 2012-01-11 | Genentech, Inc. | Antibody formulation |
SG10201502587SA (en) | 2010-03-01 | 2015-06-29 | Bayer Healthcare Llc | Optimized monoclonal antibodies against tissue factor pathway inhibitor (tfpi) |
EP3443985B1 (en) | 2010-08-23 | 2020-12-16 | XBiotech, Inc | Treatment for neoplastic diseases |
US9427477B2 (en) | 2011-05-09 | 2016-08-30 | Mayo Foundation For Medical Education And Research | Cancer treatments |
BR112014000042A2 (en) * | 2011-07-05 | 2017-02-21 | Novozymes Biopharma Dk As | composition, cell culture medium, use of a composition, and method for culturing cells |
MX356808B (en) | 2011-09-23 | 2018-06-14 | Xbiotech Inc | Cachexia treatment. |
US9592297B2 (en) | 2012-08-31 | 2017-03-14 | Bayer Healthcare Llc | Antibody and protein formulations |
EP2903610B1 (en) | 2012-10-01 | 2021-11-03 | Mayo Foundation For Medical Education And Research | Cancer treatments |
CN113134095A (en) | 2014-06-16 | 2021-07-20 | 梅约医学教育与研究基金会 | Treatment of myeloma |
US9446148B2 (en) | 2014-10-06 | 2016-09-20 | Mayo Foundation For Medical Education And Research | Carrier-antibody compositions and methods of making and using the same |
KR102210495B1 (en) * | 2014-10-06 | 2021-02-02 | 메이오 파운데이션 포 메디칼 에쥬케이션 앤드 리써치 | Carrier-antibody compositions and methods of making and using the same |
JP6921802B2 (en) * | 2015-08-18 | 2021-08-18 | マヨ ファウンデーション フォー メディカル エデュケーション アンド リサーチMayo Foundation For Medical Education And Research | Carrier Binder Composition and Methods for Making and Using It |
TW201707725A (en) | 2015-08-18 | 2017-03-01 | 美國馬友醫藥教育研究基金會 | Carrier-antibody compositions and methods of making and using the same |
TW201713360A (en) | 2015-10-06 | 2017-04-16 | Mayo Foundation | Methods of treating cancer using compositions of antibodies and carrier proteins |
US11571469B2 (en) | 2016-01-07 | 2023-02-07 | Mayo Foundation For Medical Education And Research | Methods of treating cancer with interferon wherein the cancer cells are HLA negative or have reduced HLA expression |
US11351254B2 (en) | 2016-02-12 | 2022-06-07 | Mayo Foundation For Medical Education And Research | Hematologic cancer treatments |
CA3018340A1 (en) | 2016-03-21 | 2017-09-28 | Mayo Foundation For Medical Education And Research | Methods for improving the therapeutic index for a chemotherapeutic drug |
WO2017165440A1 (en) | 2016-03-21 | 2017-09-28 | Mayo Foundation For Medical Education And Research | Methods for reducing toxicity of a chemotherapeutic drug |
US10618969B2 (en) | 2016-04-06 | 2020-04-14 | Mayo Foundation For Medical Education And Research | Carrier-binding agent compositions and methods of making and using the same |
EP4177271A1 (en) | 2016-09-01 | 2023-05-10 | Mayo Foundation for Medical Education and Research | Carrier-pd-l1 binding agent compositions for treating cancers |
US11160876B2 (en) | 2016-09-01 | 2021-11-02 | Mayo Foundation For Medical Education And Research | Methods and compositions for targeting t-cell cancers |
EP3510048A1 (en) | 2016-09-06 | 2019-07-17 | Mayo Foundation for Medical Education and Research | Methods of treating pd-l1 expressing cancer |
EP3509635A1 (en) | 2016-09-06 | 2019-07-17 | Vavotar Life Sciences LLC | Methods of treating triple-negative breast cancer using compositions of antibodies and carrier proteins |
CA3035653A1 (en) | 2016-09-06 | 2018-03-15 | Mayo Foundation For Medical Education And Research | Paclitaxel-albumin-binding agent compositions and methods for using and making the same |
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GB1546177A (en) * | 1976-11-19 | 1979-05-16 | Biokema Sa | Process for the preparation of a stable injectable solution |
JPS5822085B2 (en) * | 1977-07-19 | 1983-05-06 | 株式会社ミドリ十字 | Intravenous gamma globulin preparations |
ATE82136T1 (en) * | 1987-08-10 | 1992-11-15 | Miles Inc | PURIFIED IGM. |
-
1990
- 1990-03-13 EP EP19900905158 patent/EP0465513A1/en not_active Withdrawn
- 1990-03-13 CA CA 2049342 patent/CA2049342A1/en not_active Abandoned
- 1990-03-13 JP JP50510090A patent/JPH04504253A/en active Pending
- 1990-03-13 WO PCT/US1990/001383 patent/WO1990011091A1/en not_active Application Discontinuation
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JPH04504253A (en) | 1992-07-30 |
EP0465513A1 (en) | 1992-01-15 |
WO1990011091A1 (en) | 1990-10-04 |
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