Ulllted States Patent [19]
`Sarno et al.
`
`[54] STABLE INTRAVENOUSLY-
`ADMINISTRABLE IMMUNE GLOBULIN
`PREPARATION
`
`_
`Inventors: Maria Erlinda CI Sarno, Cerritos;
`Rodolfo Anthony Vasquez, NorWalk,
`_
`_
`_
`sau'Gee Yullg’ R1211“), Ch?ord R-_
`Graf, Lakeview Terrace, all of Calif.
`
`[4] Notice:
`
`.
`
`[73] Ass1gnee: ?laxter International Inc., Deer?eld,
`'
`This' patent issued on a continued pros-
`ecution application ?led under 37 CFR
`1.53(d), and is subject to the tWenty year
`patent term provisions of 35 U.S.C.
`154(a)(2)_
`
`.
`
`.
`
`.
`
`US005945098A
`[11] Patent Number:
`[45] Date of Patent:
`
`5,945,098
`*Aug. 31, 1999
`
`4,692,331
`4,719,290
`4,721,777
`4,764,369
`
`9/1987 Uemura et al. ......................... .. 424/85
`1/1988 Curry etal.
`530/357
`1/1988 Uemura et al. .
`530/388.15
`8/1988 Neurath et al. ......................... .. 424/89
`
`Woods et al. ......................... .. 4,876,088 10/1989 Hirao et al. ..
`
`4 877 608 10
`,
`,
`/1989 Lee et al.
`424/858
`5,177,194
`1/1993 Saras et al. ........................... .. 530/412
`
`424/85.8
`
`FOREIGN PATENT DOCUMENTS
`0187712 7/1986 European Pat‘ Off‘ '
`0196761 10/1986 European Pat. Off. .
`5347515 42978 Ia???”
`55464630 12/1980 Japan_
`59.97057 6/1984 Japan _
`
`2500076 7 1976 G
`
`.
`
`OTHER PUBLICATIONS
`Tomono et al., Vox Sang vol. 51 (2) pp. 81—86 (1986).
`Stephan, W., “Undegraded Gamma—Globulin for Intrave
`nous Therapy: ANeW Preparation of Immune Serum Globu
`lin for Intravenous Administration,” XXIVth Scienti?c Meet
`ing of the Blood Research Institute, pp. 469—478, 1971.
`Fernandes et al., “Preparation of a Stable Intravenous Gam
`ma_G1Obuhn: Process Deslgn and SCa1e_Up’” VOX Sang‘ 39:
`101—12(1980)~
`Gerber, “Stabilization of Gamma Globulin With Copper
`Complexes. possible Relevance to the Etiology of Rheuma_
`t .d Arth .t'.
`,, I
`.
`D.
`a, C
`1982
`
`n 1S’_ fit/‘lamination ‘$64868 an 01 Gerber, “Inhibition of Denaturation of Human Gamma Opper (
`
`
`Globulin by a Mixture of L—Histidine L—Cystine and
`Copper, and Its Clinical Implication in Rheumatoid Arthri_
`tiS,”Ar[hri[iS and Rheumatism) V01' 19, 3.593_601 (1976)'
`Florence et al., “Emulsion Stabilization by Non—Ionic Sur
`factants: Experiment and Theory,” J. Pharm. Pharmac.
`23;153_169 (1971)_
`Miller, G., et al., “Stabilization of Cytotoxic Antibody and
`Complement With Cysteine,” J. 0fImmun., vol. 101, No. 5,
`
`a
`
`a
`
`[21] Appl- NO-I 08/935,294
`.
`_
`[22] Flled'
`
`Sep' 22’ 1997
`t- D t
`R l t d Us A l-
`e a e
`pp lea Ion a a
`[63] Continuation of application No. 08/504,854, Jul. 20, 1995,
`abandoned, which is a continuation of application No.
`08/317,214, Oct. 3, 1994, abandoned, which is a continua-
`tion of application No. 08/178,432, Jan. 6, 1994, abandoned,
`which is a continuation of application No. 07/866,089, Apr.
`
`6, 1992, abandoned, which is a continuation of application NQ ()7/473j5547 Feb 1, 1990, abandoned
`
`6
`
`Int. Cl. ................................................... .. A61K 38/21
`[51]
`[52] US. Cl. .............................. .. 424/855; 514/2; 514/12;
`514/21; 530/387'1; 530/389'1; 530/380;
`530/390'5
`[58] Field of Search .............................. .. 424/85.5; 514/2,
`514/12> 21; 530/387-1> 389~1> 380> 3905
`_
`
`
`
`References Clted U'S' PATENT DOCUMENTS
`
`Primary Examiner—Chhaya D. Sayala
`
`3,903,262
`4,165,370
`4,168,303
`4,186,192
`
`9/1975 Pappenhagen et al. ................ .. 424/85
`8/1979 Coval .......... ..
`424/858
`9/1979 Nishida et al.
`530/387
`1/1980 Lundblad et al.
`.. 424/858
`
`Attorney) Agent) or Firm—R0IhW@11, Figg, Ernst & KHIZ,
`P-C
`
`[57]
`
`ABSTRACT
`
`128225 9128 $22,311.11; .................. .1252/221 4374 763 2/1983 Takagi .................................. .. 424/858
`
`4:384j993
`5/1983 Sam et a1_
`__ 424/85_1
`4,396,608
`8/1983 Tenold ________ __
`424/85_8
`4,439,421
`3/1984 Hooper et al.
`424/85
`4,477,432 10/1984 Hardie .................................. .. 424/85.1
`Curry et al. .......................... ..
`giurath 6t ‘111'
`7/1986 Zolton et al. .
`4,597,966
`4,613,501
`9/1986 Horowitz . . . . . .
`4 623 717 11/1986 Fernandes et a1_
`
`,
`
`,
`
`mura e a.
`
`..... ..
`
`.. 424/858
`. . . . .. 424/89
`530681
`
`muons are Stablhzed agamst aggreganon and Polymenza'
`tion and rendered isotonic With amino acid(s) and non-ionic
`detergents, polysorbate and polyethylene glycol. The immu
`noglobulins are derived from human or animal sources, or
`from hybridornag, Optional, additional stabilizers include
`Various physiologically_acceptable Carbohydrates and Salts_
`Polyvinylpyrrolidone can be used in addition to the amino
`:‘.C1d(S)'AIz;rt ?om the Tr?umglfl’ulfm Half’ the melt)?“
`1on5 are 0 . “.Wlse essen la y pro em .ree'
`é’prepara Ions
`are useful in immunotherapy and as diagnostic reagents.
`
`-
`
`-
`
`-
`
`_
`
`-
`
`4,637,834
`
`1/1987 Thurow . . . . . . . . . . . . .
`
`. . . . . . .. 514/3
`
`4,639,513
`
`1/1987 Hou et al. ............................. .. 530/387
`
`15 Claims, No Drawings
`
`Ex. 1023 - Page 1 of 5
`
`AMGEN INC.
`Exhibit 1023
`
`

`
`1
`STABLE INTRAVENOUSLY
`ADMINISTRABLE IMMUNE GLOBULIN
`PREPARATION
`
`This is a continuation of Ser. No. 08/504,854 ?led on Jul.
`20, 1995 noW abandoned, Which is a continuation of Ser. No.
`08/317,214 ?led Oct. 3, 1994, noW abandoned, Which is a
`continuation of Ser. No. 08/178,432, ?led Jan. 6, 1994, noW
`abandoned, Which is a continuation of Ser. No. 07/866,089,
`?led Apr. 6, 1992, noW abandoned, Which is a continuation
`of Ser. No. 07/473,554, ?led Feb. 1, 1990, noW abandoned.
`BACKGROUND OF THE INVENTION
`1. Technical Field
`The present invention relates to intravenously
`administrable immune globulin preparations. More
`speci?cally, the invention relates to immune serum globulin
`preparations that, aside from the immune globulin itself, are
`otherWise essentially protein free. The preparations are
`administrable by other routes as Well. The preparations also
`are useful reagents for diagnostic testing and can be supplied
`in their solution form in diagnostic test kits.
`The term immune globulin, also knoWn as Immunoglo
`bulin G, IgG and gamma globulin, encompasses both
`immune serum globulin and monoclonal immune globulins.
`Immune serum globulin is obtained from pooled plasma
`samples from either normal or hyperimmune donors and
`contains IgG antibodies to many common bacterial and viral
`infectious agents. Monoclonal immune globulins
`(monoclonal antibodies) are obtained from hybridomas.
`Immune globulin long has been used in connection With
`the prophylaxis and treatment of a variety of diseases and
`disorders. Certain patients With immunode?ciencies are
`rendered susceptible to acute and chronic infections, With
`life-threatening consequences, Which the immune system of
`the normal patient can easily combat. These patients With
`compromised immune systems are not able to produce
`normal levels of antibodies and can greatly bene?t from the
`administration of exogenous immune globulin.
`Patients With normal immune systems and antibody levels
`may require additional defense in overcoming certain
`antibiotic-resistant bacterial infections, such as those caused
`by Pseudomonas aeruainosa. Immunotherapy via the
`administration of immune globulin has become a standard
`treatment in these circumstances.
`The administration of immune globulin is perhaps most
`Widespread in the prophylaxis of What Were once highly
`common diseases. Many viral infections, such as German
`measles, measles, mumps and smallpox, for example, can be
`controlled via the Widespread administration of the appro
`priate immune globulins to children. The occurrence of
`many bacterial infections also have been controlled or
`virtually eliminated due to the Widespread administration of
`immune globulins.
`Immune serum globulins for human administration Were
`?rst produced via an alcohol fractionation method developed
`by E. Cohn et al., J. Amer. Chem. Soc. 68: 459 (1946). These
`preparations contained approximately ten to eighteen (Wt.)
`percent protein and Were relatively stable during storage at
`2 to 8° C. Although originally intended for intravenous
`administration, it Was found that untoWard reactions, includ
`ing anaphylactic shock, rendered unsafe such administration
`of the alcohol fractionated immune serum globulins. These
`products are acceptable for intra-muscular injection, hoW
`ever.
`Aparticularly preferred method for producing an immune
`globulin fraction from Cohn Fractions I+II+III, for use in the
`
`10
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`20
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`25
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`30
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`35
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`40
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`45
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`
`65
`
`5,945,098
`
`2
`present invention, is disclosed in US. Pat. No. 5,177,194
`?led concurrently hereWith and incorporated by reference
`herein.
`Although intramuscular injection of immune globulin can
`effectively raise circulating antibody levels in most patients,
`this mode of administration suffers from several disadvan
`tages. The antibodies administered in this manner diffuse
`rather sloWly into the circulation. The ultimate blood levels
`attained from a given dose varies from patient to patient, and
`local tissue injury at the injection site is a possibility.
`Moreover, only a fraction of the total immune globulin
`administered to the patient reaches the bloodstream.
`The intravenous administration of immune globulin over
`comes these disadvantages. The desired level of circulating
`antibody is reached almost instantaneously and can be
`relatively accurately controlled by control of the dosage.
`Effectively larger doses can be more quickly administered
`Without the discomfort associated With intramuscular injec
`tion.
`As mentioned earlier, the intravenous injection of certain
`immune globulin preparations has lead to severe reactions.
`These reactions are generally believed to be caused by the
`activation of complement by aggregates and/or fragments of
`immunoglobulin Which form during the preparation and/or
`storage of the immune globulin product. Thus, the safe
`administration of immune globulin requires steps to be taken
`to avoid the formation of immunoglobulin fragments and
`agglomerates and their resulting anticomplement activity.
`Immune globulins (antibodies) also ?nd Wide use in the
`diagnostics ?eld. Diagnostic test kits, intended for a single
`use, often contain a monoclonal antibody preparation as one
`of the test reagents. These antibody preparations are com
`monly provided in lyophiliZed form in order to increase their
`shelf life, and must be reconstituted just prior to use. The
`immune globulin preparations of the present invention,
`hoWever, are quite stable and are Well suited to being
`included in diagnostic test kits in their solution form.
`2. Description of the Prior Art
`The processing of immune globulin preparations in order
`to reduce anticomplement activity is documented in the
`literature. In “Preparation of a Stable Intravenous Gamma
`Globulin: Process Design and Scale Up”, Fernandes, P. M.
`and Lundblad, J. L., Vox Sang. 39: 101—112 (1980), the
`authors note that a variety of such processing methods have
`been investigated. The various techniques are (a) removal of
`high-molecular-Weight aggregates by centrifugation, (b) pH
`adjustment to 4.0, (c) ‘gentle’ fractionation techniques, (d)
`af?nity methods, (e) use of protein stabiliZers and
`enZy
`matic treatment and other chemical modi?cations. None of
`these methods has been entirely successful from either the
`purely medical or commercial vieWpoints, hoWever. For
`example, the various chemical treatments can alter the
`immunoglobulin’s physical structure and hence its activity.
`Some treatments for disassociating immunoglobulin
`agglomerates are not permanent; the agglomerates reappear
`during storage or during pH shifts to physiological condi
`tions.
`Japanese Kokai No. 60-146832 (1985) discloses that IgG
`preparations prepared via the cold ethanol fractionation and
`ammonium sulfate fractionation methods contain from 10 to
`40 percent aggregated IgG in addition to 75 IgG. These
`preparations are unsuitable for intravenous administration
`because of their anticomplement activating effects. It is
`reported that these same preparations, after treatment by
`maintenance at pH 3.7 to 4.3 at 1 to 10° C. over the course
`of 0.5 to 20 hours, no longer exhibit anticomplement activ
`
`Ex. 1023 - Page 2 of 5
`
`

`
`5,945,098
`
`3
`ity. A stabilizing compound selected from among various
`inorganic salts, sugars, proteins and organic acid salts is
`added to prevent denaturation of the immunoglobulin chain
`during the acid treatment.
`US. Pat. No. 4,719,290 (Curry et al.; Jan. 12, 1988)
`provides a reportedly intravenously-administrable gamma
`globulin preparation by stabilizing a puri?ed Cohn Fraction
`II With relatively large amounts of human serum albumin.
`The ?nal solution is formulated With the addition of sodium
`chloride and/or a carbohydrate.
`US. Pat. No. 4,396,608 (Tenold et al.; Aug. 2, 1983)
`relates to a process for stabiliZing immune serum globulin
`preparations so as to avoid aggregation. The pH of a Fraction
`II (or other Cohn fraction) ?ltrate is adjusted to 3.5—5.0 and
`the ionic strength is then adjusted (loWered) via ?ltration,
`dialysis, etc. or combinations thereof. The solution is then
`made isotonic via the addition of an amino acid, a carbo
`hydrate or a sugar.
`US. Pat. No. 4,093,606 (Coval; Jun. 6, 1978) relates to
`fractionation techniques Whereby a suspension of Cohn
`Fraction II or II+III plasma protein is reportedly rendered
`stable for intravenous injection. Preparations are formulated
`With glycine, albumin and a non-ionic surfactant such as one
`of the polysorbates.
`Although albumin is knoWn for its ability to stabiliZe
`proteins in solution, its stabiliZing properties diminish With
`time. Other factors also detract from the use of albumin. The
`presence of serum-derived albumin in a preparation may
`detract from its overall purity, as it is noW Well knoWn that
`many contaminants, including proteins and infectious
`agents, can be neither easily detected nor removed With
`certainty in a cost effective manner. The presence of albumin
`also can mask the presence of immunoglobulin fragments in
`the immune serum. If such fragments are present, yet
`undetected, the presence of undesirable enZymes Which
`produce such fragments also may go undetected. Thus, a
`need exists for improved stabiliZed immune globulin prepa
`rations Which are albumin-free.
`
`SUMMARY OF THE INVENTION
`
`In one of its aspects the present invention relates to a
`stable, intravenously-administrable immune globulin prepa
`ration comprising an aqueous solution of immune globulin,
`an amount of an amino acid effective to impart a
`physiologically-acceptable tonicity to the preparation and to
`maintain the immune globulin in monomeric form, and a
`physiologically-acceptable amount of a non-ionic detergent
`Wherein, apart from the immune globulin itself, the prepa
`ration is essentially protein-free.
`In another aspect, the present invention relates to a stable,
`intravenously-administrable immune globulin preparation
`comprising an aqueous solution of immune globulin, a
`physiologically acceptable amount of polyvinylpyrrolidone
`effective to maintain said immune globulin in monomeric
`form, an amount of a carbohydrate or a physiologically
`acceptable salt or an amino acid effective to impart a
`physiologically-acceptable tonicity to the preparation, and a
`physiologically-acceptable amount of a non-ionic detergent
`Wherein again the preparation is essentially protein-free
`apart from the immune serum globulin.
`The present invention advantageously provides a storage
`stable liquid product, thus eliminating the inconvenience
`and expense of lyophiliZation and reconstitution. The ionic
`strength, pH and protein concentration can be optimiZed
`during processing, and the ?nal product can be stored at
`ambient temperatures or under refrigeration. The stabiliZers
`
`4
`are non-toxic and present in minimal amounts, and they do
`not alter the native biological form or function of the
`immunoglobulin. PolymeriZed immunoglobulin does not
`form during storage; thus, the preparations are Well suited
`for intravenous administration. The preparations also are
`useful as stable immune globulin reagents for use in diag
`nostic applications.
`
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`
`DETAILED DESCRIPTION OF THE
`INVENTION
`
`The immune globulin preparations according to the
`present invention are prepared from any starting materials
`based on native or monoclonal immunoglobulin G. The
`teachings herein are applicable to immunoglobulins derived
`from both native and monoclonal sources. Most frequently
`the preparations Will be prepared from gamma globulin
`containing products produced by the alcohol fractionation
`and/or ion exchange and af?nity chromatography methods
`Well knoWn to those skilled in the art. Puri?ed Cohn Fraction
`II is commonly used and is the starting point for the
`preparations disclosed herein unless otherWise noted.
`As accepted in the art, the starting Cohn Fraction II paste
`is about 95 percent pure IgG and is comprised of each of the
`four subclasses of IgG, namely, IgG1, IgG2, IgG3 and IgG4.
`The different subclasses are present in Fraction II in approxi
`mately the same ratio as they are found in the pooled human
`plasma from Which they are obtained. The Fraction II must
`be further puri?ed before formulation into an administrable
`product, and this can be accomplished via any of a number
`of knoWn procedures so that the ?nal immune serum globu
`lin product does not contain impurities, such as IgG poly
`mers and/or fragments, that possess anticomplement activi
`ties.
`For example, the Fraction II paste can be dissolved in a
`cold puri?ed aqueous alcohol solution and impurities
`removed via precipitation and ?ltration. FolloWing the ?nal
`?ltration the immunoglobulin suspension is dia?ltered using
`ultra?ltration membranes having a nominal molecular
`Weight limit of 2 100,000 daltons to remove the alcohol. The
`solution is either concentrated or diluted to obtain the
`desired protein concentration and can be further puri?ed by
`techniques Well knoWn to those skilled in the art.
`Aphysiologically acceptable amount of a non-ionic deter
`gent is added to the immunoglobulin solution in accordance
`With the present invention. By ‘physiologically acceptable’
`it is meant that the amount added is a stabiliZing, non-toxic
`amount loW enough to avoid any adverse toxic reactions as
`derived from knoWn LD50 values and/or published literature
`pertinent to the potential toxicity of the particular additive.
`It is advantageous to use the least amount necessary to
`produce the desired stabiliZation. The detergent reduces the
`tendency of the immunoglobulin molecules to come out of
`solution. Suitable detergents (surfactants) include the partial
`esters of sorbitol and polyoxyethylene oxides of long chain
`fatty acids knoWn as the polysorbates, alkylpolyethoxyetha
`nols and alkarylpolyethoxyethanols. Polysorbate 80, com
`mercially available as TWeenTM 80 from Atlas Chemical
`Industries, is most preferred. The non-ionic detergents are
`advantageously used in minimal amounts and preferably
`beloW about 0.01 percent (W/v). It has been found that from
`0.0005 to 0.01 percent is useful, and ?nal products contain
`ing about 0.002 percent (W/v) of non-ionic detergent are
`preferred When the immune serum globulin content is 5
`percent (W/v). At higher immune serum globulin
`concentrations, for example 10 percent, the detergent level
`(i.e. detergent concentration to protein ratio) can be reduced
`
`Ex. 1023 - Page 3 of 5
`
`

`
`10
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`15
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`20
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`30
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`35
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`40
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`5
`even further. At these higher protein concentrations the
`immunoglobulin molecules appear to stabilize themselves.
`The optimal detergent content is obtained by formulation
`experiments Well knoWn to those skilled in the art. In any
`case, it appears that a desirable detergent concentration
`provides a balance of hydrophilic and hydrophobic proper
`ties to the protein solution so that the immunoglobulin is less
`prone to denature. The optimal detergent concentration Will
`be the minimal amount that Will prevent particles from
`coming out of solution, Will not cause toxic effect and Will
`not adversely alter the efficacy of the ?nal product under the
`selected storage conditions.
`An amount of one or more amino acids effective to impart
`a physiologically-acceptable tonicity (i.e. about 300 mOsm/
`l) to the preparation and to maintain said immune serum
`globulin in monomeric form also is added to the solution.
`Preferred amino acids include glycine, lysine, arginine and
`phenylalanine, With glycine being particularly preferred. A
`glycine concentration of about 0.2 M generally is preferred.
`The IgG solution can further comprise polyvinylpyrroli
`done (“PVP”) at certain pH ranges, typically pH 4 to pH 7,
`in addition to or in place of the amino acid(s) as an agent for
`preventing polymeriZation and/or particulation of the
`gamma globulin. PVP alone typically does not provide long
`term stabiliZation of immunoglobulins, but PVP does con
`tribute to effective long term stabiliZation in the preparations
`of this invention. The amount of PVP useful in the prepa
`rations is limited by its toxicity and insolubility at high
`concentrations. From 0.1 to 1 percent (W/v) of PVP K-15 (or
`other Water soluble grade) is useful to prevent particulation.
`Amounts Within this range are soluble and Well beloW toxic
`levels.
`The addition of PVP does not provide an isotonic solution
`(except When the PVP is added at unacceptably high levels).
`Therefore, PVP generally is added to the IgG solutions in
`combination With other additives such as amino acids,
`physiologically-acceptable salts or carbohydrates Which can
`be used to adjust for isotonicity. Useful carbohydrates
`include sugars such as dextrose, mannose, trehalose,
`galactose, dextran, fructose, lactose, sucrose, maltose, man
`nitol and sorbitol. Non-reducing carbohydrates are pre
`ferred. Almost any physiologically acceptable salt, such as
`sodium chloride, sodium acetate, sodium citrate, etc., can be
`used to adjust the tonicity of the preparations. Examples
`include the alkali metal, alkaline earth metal, ammonium
`and phosphate salts as Well as various organic salts
`(including but not limited to oxalate, valerate, oleate,
`laurate, borate, benZoate, lactate, tosylate, citrate, maleate,
`fumarate, succinate and tartrate). Sodium chloride and
`sodium citrate are preferred salts. The speci?c amount of
`any particular tonicity-adjusting agent(s) needed to provide
`an isotonic solution Will vary from preparation to prepara
`tion. The artisan Will readily determine appropriate amounts,
`hoWever.
`It has been found that the pH of the immune globulin
`preparation is an important factor relative to the IgG mono
`mer content of the ?nal product. In the preparations of the
`present invention the pH can range from 4.0 to 7.4. Within
`this range, a loWer pH is desired, hoWever, to ensure
`maximum monomer content. A pH of 4.2105 is preferred
`for a 5 percent immune serum globulin preparation. Ten
`percent preparations, hoWever, are most stable at a pH of
`5 .2102. Optimal pH is obtained by formulation techniques
`Well knoWn to those skilled in the art. For example, optimal
`pH can be determined from siZe exclusion chromatography
`determinations as Well as heat stability data and anticomple
`ment titers of the various preparations under differing pH
`conditions.
`
`55
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`60
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`5,945,098
`
`6
`Immune globulin solutions, including immune globulin
`solutions useful as starting materials for the present
`preparations, often include minor amounts of poly(ethylene
`glycol) (“PEG”) as a stabiliZer. While PEG alone cannot
`provide a preparation as stable as those described herein, its
`presence is believed to be important to the overall stability
`of any immune globulin solution, including those of the
`present invention. Thus, if PEG is not already present in the
`starting source of immune globulin, a small amount
`(typically less than 0.2 gram %) should be included in the
`preparations of the invention.
`It Will be apparent that the appropriate concentrations of
`the various stabiliZers in the present immune globulin com
`positions Will vary With protein concentration. Normal
`variations among pooled sera also Will cause adjustments in
`stabiliZer requirements. Those skilled in the art Will readily
`determine acceptable stabiliZer levels based on the teachings
`herein.
`The invention is illustrated by the folloWing Examples.
`All percentages are by Weight unless otherWise noted.
`
`EXAMPLE I
`
`Glycine at concentrations of 0.210.1M is added to aque
`ous IgG solutions puri?ed according to the method taught in
`US. Pat. No. 5,177,194 from Cohn Fr. I+II+III, or other
`more crude IgG sources Which contain residual amounts of
`polyethylene glycol, usually less than 0.2 g %. The protein
`concentration is then adjusted to approximately 5% either by
`dissolution and dilution of a puri?ed IgG precipitate or
`concentration through the use of tangential ?oW ultra?ltra
`tion and other suitable means of concentrating proteins. The
`pH is adjusted to 5 .0110 and Polysorbate 80 (TWeenTM 80)
`is then added to a 0.003:0.001% level. The IgG solution is
`sterile ?ltered and ?lled into ?nal containers for use.
`
`EXAMPLE II
`
`Glycine at concentrations of 0.210.1M is added to aque
`ous IgG solutions puri?ed from Cohn Fr. II, Cohn Fr. III
`?ltrate, or other more crude IgG sources. The protein con
`centration is then adjusted to approximately 10% and the pH
`is adjusted to 6011.0. Polysorbate 80 (TWeenTM 80) is then
`added to a 0.007:0.003% level. The IgG solution is sterile
`?ltered and ?lled into ?nal containers for use.
`
`EXAMPLE III
`
`Glycine at concentrations of 0.210.1M, PEG at concen
`trations of 0.01% and polysorbate at concentrations of
`approximately 0.001% are added to an aqueous solution of
`puri?ed monoclonal antibody derived from hybridomas. The
`concentration of the antibody is approximately 2 mg/ml and
`the solution is kept at pH 5.2 using 10 mM sodium citrate or
`sodium phosphate as buffer. This preparation can be used as
`a diagnostic reagent or can be sterile ?ltered and ?lled into
`?nal containers for intravenous or intramuscular adminis
`tration.
`
`EXAMPLE IV
`
`Glycine at concentrations of 0.210.1M, and polyvinylpyr
`rolidone at concentrations ranging from 0.1 to 1% is added
`to aqueous IgG solutions puri?ed from Cohn Fr. II, Cohn Fr.
`III ?ltrate, or other more crude IgG sources Which contain
`residual amount of PEG (typically less than 0.2%). The
`protein concentration is then adjusted to the desired value,
`usually 5 or 10%, by knoWn dilution or concentration
`techniques such as ultra?ltration. The pH is adjusted to
`
`Ex. 1023 - Page 4 of 5
`
`

`
`5,945,098
`
`7
`5011.0. Polysorbate 80 (TWeenTM 80) is then added to a
`concentration that Will not cause particulation in the
`solution, typically, 000310.001 for 5% protein solution or
`0.007%:0.003% for a 10% protein solution. The IgG solu
`tion is sterile ?ltered and ?lled into ?nal containers for use.
`
`EXAMPLE V
`
`Glycine at concentrations of 0.210.1M and carbohydrates
`such as trehalose, glucose, mannitol, hydroXyethyl starch,
`carboXy methyl cellulose, hydroXymethyl starch, maltose,
`glycerol, lactose etc. at concentrations of 0.1 to 3% is added
`to aqueous IgG solutions puri?ed from Cohn Fr. II, Cohn Fr.
`III ?ltrate, or other more crude IgG sources Which contain
`residual amount of PEG, typically less than 0.2 g %. The
`protein concentration is then adjusted to the desired
`concentration, usually 5 or 10%, by knoWn dilution or
`concentration techniques such as ultra?ltration. The pH is
`adjusted to 5.0110. Polysorbate 80 (TWeenTM 80) is then
`added to a concentration that Will not cause particulation in
`the solution, typically 0.003:0.001% for a 5% protein
`solution or 0.007:0.003% for a 10% protein solution. The
`IgG solution is sterile ?ltered and ?lled into ?nal containers
`for use.
`Although the invention has been described in connection
`With certain preferred embodiments, it is not so limited.
`Variations Within the scope of the claims Will be apparent to
`those skilled in the art.
`We claim:
`1. A storage stable, intravenously-administrable immune
`globulin preparation comprising an aqueous solution of
`immune globulin, from about 0.1 M to about 0.3 M glycine,
`from about 0.0005% (W/v) to about 0.01% (W/v)
`polysorbate, and less than about 0.2 gram % PEG, Wherein
`the preparation is essentially protein-free apart from said
`immune globulin.
`2. An immune globulin preparation according to claim 1
`Wherein said polysorbate detergent is polysorbate 80.
`3. An immune globulin preparation according to claim 2
`Wherein the immune globulin concentration is about 5
`percent (W/v) and the concentration of polysorbate 80 is
`about 0.002 percent (W/v).
`4. An immune globulin preparation according to claim 1
`Wherein the immune globulin concentration is about 5
`
`10
`
`15
`
`20
`
`25
`
`30
`
`35
`
`40
`
`8
`percent (W/v) and the concentration of polysorbate is about
`0.002 percent (W/v).
`5. An immune globulin preparation according to claim 1
`having a glycine concentration of about 0.2 M.
`6. An immune globulin preparation according to claim 1
`having an immune globulin concentration of about 5% (W/v)
`and a pH of about 4 to 7.4.
`7. An immune globulin preparation according to claim 1
`having an immune globulin concentration of about 10%
`(W/v) and a pH of about 5 to 7.4.
`8. An immune globulin preparation according to claim 1
`further comprising a carbohydrate or a physiologically
`acceptable salt.
`9. An immune globulin preparation according to claim 8
`Wherein said carbohydrate is selected from the group con
`sisting of dextrose, mannose, trehalose, galactose, deXtran,
`fructose, sucrose, maltose, mannitol and sorbitol.
`10. An immune globulin preparation according to claim 1
`Wherein the immune globulin is derived from a monoclonal
`source.
`11. An immune globulin preparation according to claim 1
`Wherein the immune globulin concentration is about 10
`percent (W/v) and the polysorbate detergent concentration is
`from about 0.0005 to less than about 0.002 percent (W/v).
`12. A storage stable, intravenously-administrable immune
`globulin preparation according to claim 1 Wherein the
`polysorbate is present from about 0.002% (W/v) to about
`0.01% (W/v).
`13. Amethod for preparing a storage stable intravenously
`administrable immune globulin preparation comprising
`imparting a physiological tonicity to an aqueous immuno
`globulin solution by adding from about 0.1 M to about 0.3
`M glycine, from about 0.0005% (W/v) to about 0.01% (W/v)
`polysorbate, and less than about 0.2 gram % PEG, thereto,
`said preparation being essentially protein-free apart from
`said immune globulin.
`14. A method according to claim 13 Wherein the polysor
`bate is added in an amount from about 0.002% (W/v) to
`about 0.01% (W/v).
`15. A method according to claim 13 further comprising
`the addition of a physiologically acceptable amount of
`polyvinylpyrrolidone effective to maintain said immune
`globulin in monomeric form.
`
`*
`
`*
`
`*
`
`*
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`*
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`Ex. 1023 - Page 5 of 5