`
`a» UK Patent Application «»GB «12293 100 «A
`
`(43) Date of A Publication 20.03.1996
`
` (51)
`Application No 9418609.5
`iNT CLE
`A6G1K 47/02 9/08
`
`
`
` Date of Filing 15.09.1994
`(52) UKCL (Edition O)
`
`ASB BAB B111 8821 B823
`
`Applicantis)
`
`Medeva EuropeLimited
`(56) Documents Cited
`
`EP 0568726 A2
`EP 0332826 A1
`WO 94/21298 A1
`(Incorporatedin the United Kingdom)
`WPI Accession No:94-307582/38 & JP-6234659-A
`
`10 St James's Street, LONDON, SW1A 1EF,
`United Kingdom
`
`
`
`(74) Agent and/or Address for Service
`J AKemp & Co
`
`14 South Square, Gray's Inn, LONDON, WC1R 5LX,
`
`
` United Kingdom
`
`(54) Pharmaceutical compositions with deuterium oxide
`
`(72)
`
`Inventor(s)
`tan George Stephen Furminger
`Philip James Sizer
`
`(58)
`
`Field of Search
`UK CL (Edition N ) A5B BAB BLB BNB
`INT CL® AG1K 47/02 47/16 47/18 47/26 47/36
`ONLINE: WP1, CAS-ONLINE
`
`(57) A pharmaceutical composition comprises a solution or suspension of a biological agent in a diluent
`comprising D20, which solution or suspension contains sufficient amounts of a sugar, phosphate, glutamic
`acid or a physiologically acceptable salt thereof and, optionally, a non-toxic protein, a peptone or a mixture of
`two or more amino acids besides glutamic acid to enhancethestability of the agent. Typically, the
`composition is a suspensionof a live attenuated poliovirus or an inactivated poliovirus of each of poliovirus
`types 1, 2 and 3. Preferably the solution or suspension contains stabilising amounts of sucrose, KH2PO,,
`K2HPQ,, L-glutamic acid or sodium glutamate and human serum albumin (SPGA).
`
`At least one drawing originally filed was informal and the print reproduced hereis taken fromalater filed formal copy.
`
`
`
`VOOL€6¢2gD
`
`
`
`
`
`
`
`1
`
`2293100
`
`PHARMACEUTICAL COMPOSITIONS
`
`The present
`
`invention relates to stabilised pharma-
`
`ceutical compositions and to their preparation.
`
`Biological agents such as proteins, nucleic acids and
`live attenuated or inactivated viruses canbe formulated ina
`
`number of ways for pharmaceutical use. They may be formulated
`
`as aqueous solutions or dispersions. Such solutions or
`
`dispersions should, however, have a shelf-life of several
`
`months or, better,
`a year or more to take account of the time
`that may elapse between manufacture and use.
`:
`
`10
`
`It is impossible to store solutions or dispersions of
`
`many biological agents at room temperature for prolonged
`
`periods of time without deterioration. This problem is overcome
`
`in some instances by freeze-drying to form a lyophilisate. The
`
`15
`
`lyophilisate then has to be reconstituted prior to use which
`
`can itself be a difficulty. Alternatively, solutions or
`
`dispersions may be stored at a reduced temperature such as ina
`
`refrigerator in order to reduce degradation.
`
`We have now found that solutions or dispersions of
`
`20
`
`biological agents can be better stabilised by using deuterium
`
`oxide as the principal diluent in combination with a number of
`
`other components. Especially,
`
`live virus vaccines can be
`
`better stabilised.
`
`Accordingly,
`
`the present invention provides a
`
`25
`
`pharmaceutical composition which comprises a solution or
`
`suspension of a biological agent
`
`in a diluent comprising D,,
`
`which solution or suspension contains sufficient amounts of a
`
`sugar, phosphate, glutamic acid or a physiologically acceptable
`
`salt thereof and, optionally, a non-toxic protein, a peptone or
`
`
`
`-
`
`2
`
`-
`
`a mixture of two or more amino acids besides glutamic acid to
`
`enhance the stability of the agent.
`
`The invention can be applied to biological macromolecules
`
`such as proteins and nucleic acids. It may be applied to
`
`recombinant proteins. The proteins may be cytokines, enzymes or
`
`antibodies such as interferons,
`
`interleukins, antigens and
`
`enzymes. The protein may be erythropoetin, blood clotting
`factor VIII,
`insulin, human growth hormone or another growth
`
`factor. The nucleic acid may be DNA or RNA.
`
`10
`
`The invention can also be applied to viruses,
`
`in
`
`particular to live attenuated or inactivated viruses. Such
`viruses may be used in vaccines. They are not therefore
`
`virulent.
`
`The invention is particularly applicable to live
`
`virus vaccines.
`
`15
`
`The virus may be a RNA virus or a DNA virus. The virus
`
`may be influenza virus or poliovirus. The invention can thus be
`applied to live attenuated or inactivated poliovirus types 1,
`2
`or 3. A preferred composition of the invention is a suspension
`of a live attenuated poliovirus or of an inactivated poliovirus
`
`20
`
`of each of poliovirus types 1,
`
`2 and 3.
`
`Any vaccine strain of poliovirus may be employed. The
`
`strain may therefore be a Sabin strain type 1,
`
`2 or 3
`
`poliovirus. Sabin strains are live attenuated polioviruses.
`Further, attenuated polioviruses can be constructed by genetic
`
`25
`
`for example by genetic mutation of the 5’ non-coding
`mutation,
`region of the poliovirus genome. Alternatively,
`the invention
`can be applied to Salk type 1,
`2 and 3 polioviruses. Salk
`
`vaccines are poliovirus vaccines containing inactivated
`
`poliovirus.
`
`
`
`
`
`-
`
`3
`
`-
`
`The sugar in the pharmaceutical composition of the
`
`invention is typically a mono-, di-saccharide or tri-~saccharide
`
`such as glucose, fructose, sucrose,
`
`lactose, maltose or
`
`cellobiose Preferably it is sucrose.
`
`The phosphate is typically an orthophosphate. It may be
`
`potassium dihydrogen orthophosphate, KH,PO,, and/or dipotassium
`
`hydrogen orthophosphate, K,HPO,. The phosphate can act as a
`
`buffer, for example to ensure that a solution or suspension
`
`according to the invention has a pH of from 6.2 to 7.2 such as
`
`10
`
`from 6.5 to 7.0.
`
`:
`
`Preferably the glutamic acid is L-glutamic acid. The
`
`preferred salt is the monosodium salt.
`
`The optional non-toxic protein, peptone or mixture of two
`
`or more amino acids other than glutamic acid is generally
`
`15
`
`albumin. The albumin can be bovine serum albumin or preferably
`
`human serum albumin.
`
`The biological agent is dissolved or dispersed ina
`
`diluent comprising D,O. The diluent thus consists essentially
`
`of D,O. D,O is the principal diluent. Minor amounts of other
`
`20
`
`physiologically acceptable diluents miscible with D,O may be
`
`present,
`
`typically H,0O. For example, another diluent such as
`
`H,0 may be present in an amount of up to 30%,
`
`for example up to
`
`20%, by weight of the composition; for example from 1, or from
`
`5,
`
`to 10, or to 15%, by weight.
`
`25
`
`The amount of the biological agent varies depending upon
`
`the nature of the agent and the desired final concentration of
`
`the agent. The agent may be present in the pharmaceutical
`
`composition of the invention in an amount of from 1 to 20% by
`
`weight,
`
`for example from 5 to 10% by weight.
`
`
`
`- 4
`
`-
`
`In the case of viruses,
`the amount present may be
`represented by a titre. A Suitable titre may therefore be from
`1085 to 107° per ml, for example 106-§ to 10’* per ml and more
`especially about 10° per ml. Such titres are particularly
`appropriate in the case of a poliovirus.
`.
`An advantage of the present
`invention is that less of an
`active agent may be provided ina composition than would
`otherwise be the case. An excess or "overage" of an agent may
`be initially provided ina composition to take account of
`degradation of the active agent
`in the period of time between
`manufacture of the composition and use. As the present
`compositions exhibit enhanced stability,
`this overage can be
`
`reduced.
`The amount of D,O in the composition may vary depending
`for example,
`the amount of biological agent that is
`upon,
`present. Typically the amount of D,0 is from 70 to 95% by
`weight of the composition,
`for example from 75 to 90% or from
`75 to 85%. The D,O itself has an effect in increasing the
`
`10
`
`15
`
`20
`
`25
`
`stability of the compositions of the invention.
`(c)
`Sufficient amounts of
`(a) a sugar,
`(b) phosphate,
`glutamic acid or a physiologically acceptable salt thereof and,
`optionally,
`(d) a non-toxic protein, a peptone or a mixture of
`two or more amino acids besides glutamic acid are required so
`that the stability of the agent
`in the composition is enhanced.
`Suitable stabilising amounts of each component are:
`(a)
`0.5 to 3%, preferably 0.8 to 1.5%, by weight such as
`
`about
`1% by weight;
`0.5 to 1.5%, preferably 0.7 to 1%, by weight such as
`
`(b)
`
`about 0.8% by weight;
`
`
`
`-
`
`Ss
`
`-
`
`(c)
`
`0.2 to 1%, preferably 0.3 to 0.5%, by weight such as
`
`about 0.4% by weight; and
`
`(d)
`
`0.5 to 3%, preferably 1 to 2%, by weight such as about
`
`1.5% by weight.
`
`When (¢c)
`
`is glutamic acid, preferably a composition of
`
`the invention contains a weight ratio of
`
`(a): (b):(c):optionally
`
`(d) of about 1:0.76:0.37:0.46. This ratio especially applies
`
`when (a)
`
`is sucrose,
`
`(b)
`
`is (b’)KH,PO, and (b")K,HPO,,
`
`(c)
`
`is L-
`
`glutamic acid and (d)
`
`is human serum albumin.
`
`In this event,
`
`10
`
`the preferred weight ratio of
`
`(a): (b’):(b"):(c):optionally (d)
`
`is about 1:0.40:0.36:0.37:0.46.
`
`When (c)
`
`is sodium glutamate, preferably a composition of
`
`the invention contains a weight ratio of
`
`(a): (b):(c):
`
`optionally (d) of about 1:0.76:0.49:0.46. This ratio
`
`15
`
`especially applies when (a)
`
`is sucrose,
`
`(b)
`
`is (b’)KH,PO, and
`
`(b") K,HPO,,
`
`(c)
`
`is sodium glutamate and (d)
`
`is human serum
`
`albumin.
`
`In this event,
`
`the preferred weight ratio of
`
`(a): (b‘)}:(b"):(c): optionally (d)
`
`is about
`
`1:0.40:0.36:0.49:0.46.
`
`20
`
`It is believed that a synergistic effect may be operating
`
`to increase the stability of the compositions of the invention.
`
`Preferably,
`
`therefore,
`
`(a)
`
`to (c) and optionally (d) and,
`
`indeed,
`
`the D,O and (a)
`
`to (c) and optionally (d) are present
`
`in respective amounts which provide a synergistic stabilising
`
`25
`
`effect.
`
`A particularly useful composition according to the
`
`invention comprises a biological agent dissolved or dispersed
`
`ina D,0 SPG or SPGA solution.
`
`D,0 SPG and SPGA solutions are:
`
`
`
`SPG_ solution
`
`a solution in D,O of:
`
`1.09%,
`
`for
`
`example
`
`1.085%,
`
`w/v
`
`sucrose,
`
`0.43%,
`
`for
`
`example
`
`0.432%,
`
`w/v
`
`KH,PO,,
`
`0.39%,
`
`for
`
`example
`
`0.392%,
`
`w/v
`
`K,HPO,,
`
`and
`
`0.40%,
`
`for
`
`example
`
`0.403%,
`
`w/v
`
`L-glutamic
`
`acid,
`
`er 0.53%,
`
`for
`
`example
`
`0.539%,
`
`w/v sodium glutamate;
`
`SPGA solution
`
`10
`
`a solution in D,O of:
`
`1.
`
`for
`
`example
`
`1.085%,
`
`w/v
`
`sucrose,
`
`for
`
`example
`
`0.432%,
`
`w/v
`
`KH,PO,,
`
`0.39%, for
`
`example
`
`0.392%,
`
`w/v
`
`K,HPO,,
`
`0.40%, for
`
`example
`
`0.403%,
`
`w/v
`
`L-glutamic
`
`acid or
`
`0.53%,
`
`for example
`
`glutamate,
`w/v sodium
`0.529%,
`0.50% w/v human serum albumin.
`
`and
`
`15
`
`A SPG solution may alternatively be defined as 0.218 M
`
`sucrose, 0.0038 M KH,PO,,
`
`0.0072 M K,HPO, and 0.0049 M
`
`monosodium glutamate in D,O. Similarly,
`
`a SPGA solution may be
`
`20
`
`defined as 0.218 M sucrose,
`
`0.0038 M KH,PO,,
`
`0.0072 M K,HPO,,
`
`by weight albumin in D,0.
`0.0049 M monosodium glutamate and 1%
`A composition of the invention may be a single-strength
`D,O SPG or SPGA solution as defined above. Alternatively,
`It may be
`may be above or below a single-strength solution.
`for example from
`times single-strength,
`from a half to five
`one to four times single-strength. A D,O SPG or SPGA solution
`
`it
`
`25
`
`of variable strength may thus be present.
`A composition of the invention may contain additional
`components. For example, a poliovirus vaccine may contain wine
`
`
`
`
`
`-7-
`
`or more antibiotics. Typically,
`
`the vaccine may contain trace
`
`amounts of polymixin and/or neomycin.
`
`The poliovirus vaccine
`
`will necessarily contain live attenuated strains of all three
`
`types of poliovirus or inactivated strains of all three types
`
`of poliovirus.
`
`A composition of the invention is prepared according to
`
`the invention by a process which comprises dissolving or
`
`dispersing the biological agent in a D,0 solution of the said
`
`amounts of the sugar, phosphate, glutamic acid or salt thereof
`
`10
`
`and, optionally, non-toxic protein, peptone or mixture of two
`
`or more amino acids other than glutamic acid.
`
`This may be achieved in any appropriate fashion.
`
`In the
`
`case where the biological agent is a poliovirus,
`
`the poliovirus
`
`is first grown in tissue culture such as in human diploid cells
`
`i5
`
`or monkey kidney cells. The tissue culture fluid is harvested
`
`and sterilised through a filter. Concentrates obtained in this
`
`way of poliovirus strains of each of types 1,
`
`2 and 3 can then
`
`be blended together. Then the resulting blend is dissolved or
`
`dispersed in the D,O solution according to the invention.
`
`The
`
`20
`
`D,O solution can first be sterilised, for example by passing
`
`through a sterilising filter.
`
`The solution or dispersion obtained by the process of the
`
`invention is generally sterilised by passing through a
`
`sterilising filter. The sterilised solution or dispersion may
`
`25
`
`then be dispensed into containers, glass or plastic, which are
`
`then sealed. The containers are typically unit-dose or multi-
`
`dose containers such as ampoules or vials.
`
`These containers can be stored under appropriate
`
`conditions. Depending upon the nature of the biological agent,
`
`
`
`-
`
`8
`
`-
`
`they may be stored at 4 to 8°C such as in a refrigerator or at
`
`room temperature such as on the shelf. The solutions or
`
`dispersions within the containers do, however, exhibit enhanced
`
`stability and so may be stored for longer than previously under
`
`the same conditions as were used previously. Alternatively,
`
`they may not need to be stored in a refrigerator when previous
`
`solutions were so stored.
`
`The following Examples illustrate the invention.
`
`In the
`
`accompanying drawings, Figures 1,
`
`2 and 3 are graphs of mean
`
`10
`
`log,, titre of poliovirus in H,O and in D,O at day 0 and at day
`
`7 when no stabiliser is added (Figure 1), when SPGA is added as
`
`a stabiliser (Figure 2) and when MgCl, is added as a stabiliser
`
`(Figure 3).
`
`15
`
`Example 1: Preparation of diluent solutions
`
`The following diluent solutions were prepared. D,0 was
`
`from Sigma, product no. D-4501, stated to be 99.9% D. MgCl, was
`
`from Fisons, product no. M/0600/53. Albumin was human albumin
`
`20
`
`20% B.P.
`
`(British Pharmacopoeia). No antibiotics were added to
`
`the solutions. The solutions were sterilised by passing through
`
`a sterilising filter.
`
`s.s.
`
`= single strength and 4x = four
`
`times single strength.
`
`(a)
`
`4x_SPG
`
`25
`
`KH,PO,
`K,HPO,
`
`Sucrose
`
`L-Glutamic acid
`
`Water for Injections (WFI.
`
`«r DO
`
`0.173 g
`0.157 g
`
`0.434 g
`
`Q0.161g
`
`to 10 ml
`
`
`
`
`
`(b)s.s.SPGA
`
`KH,PO,
`
`K,HPO,
`
`Sucrose
`
`L-Glutamic acid
`
`Albumin (added after pH adjustment
`
`and sterilisation)
`
`0.432 g
`
`0.392 g
`
`1.085 g
`
`0.403 g
`
`7.5
`
`ml
`
`WFI or DjO... . ee ee ees to 100 ml
`
`4M KOH was added to adjust the pH to 6.6.
`
`10
`
`(c)
`
`1M_MgCl,
`
`20.3g of MgCl,.6H,0 was dissolved in 100ml of WFI or D,O
`
`(ad)3MMgCl,
`
`15
`
`6.1g of MgCl,.6H,0 was dissolved in 10m1l of WFI
`
`Example 2: Preparation of Formulations
`
`Formulations of poliovirus type 3 Sabin strain were
`prepared. A target titre of 10°? per 0.15ml1 dose was required
`which corresponded to a titre of 10° per ml. A batch of the
`poliovirus was grown in human diploid cells in culture toa
`titre of approximately 10°°°., The culture was sterilised by
`passing through a sterilising filter.
`The following
`
`formulations were prepared:
`
`20
`
`25
`
`
`
`
`
`1. No Stabiliser
`
`Type III virus
`
`
`WFI
`
`
`
`
`Type III virus
`
`WFI
`
`D,0
`
`3. SPGA
`
`Type III virus
`
`4xSPG (made in WFTI)
`
`Albumin
`
`s.s. SPGA(made in WFI)
`
`4. SPGA + D,0O
`
`Type III virus
`
` No Stabiliser + D,0
`
`5. MgCl,
`
`4xSPG(made in D,0)
`
`Albumin
`
`s.s. SPGA(made in D,0O)
`
`Type III virus
`
`3M MgCl,(made in
`
`1M MgCl,(made in
`
`€. MgCl, + D,O0
`
`Type III virus
`
`3M MgCl,(made in
`
`1M MgCl,(made in
`
`1M MgCl,(made in
`
`Formulations 3 and 4 were in fact single strength SPGA.
`
`Thorough mixing of the blends was ensured with minimum
`
`10
`
`agitation. Solutions containing D,O were opened to the air for
`
`as short a time as possible to prevent exchange with hydrogen
`
`ions from atmospheric H,O. For each formulation,
`
`the solutions
`
`were mixed in the order indicated in the Table. Using an
`
`Eppendorf repeater pipette fitted with a 50ml syringe filler,
`
`15
`
`the formulations were aliquoted into 1ml samples in bijou
`
`bottles, 80 samples per formulation. Each of the formulations
`
`
`
`- 11 _
`
`was sampled at day 0 and stored at 4°C, 37°C and 45°C for
`
`sampling on future days as below:
`
`
`
`10
`
`The samples were then kept at
`
`-20°C until ready for
`
`analysis. Virus titres were determined by an Infectivity Titre
`
`test. The Infectivity Titre test complies with WHO TRS800. The
`
`results are as follows:
`
`15 Day 0
`
`Sample
`Description
`
`Total Virus Content Mean Titre (95%
`(TCID,,/ml)
`
`Fiducial Limits) Ll. No Stabiliser
`
`107°, 10°57, 108%
`*
`
`
`
` 2. No Stabiliser 10° 5*, 10°33, 10%? 10°: *
`
`
`+ D,O
`*
`(10-78-10 °°)
`3. SPGA
`10739, 1064, 107%
`10’7°°?
`(10° °94-107-?7)
`10’7-°°
`(10° 8&-197-35)
`108-85
`(10°:74-10°% 9?)
`10° °°
`(10°:75-107°°3)
`10°?
`(107-99-10°:*7)
`
`4. SPGA + D,0O
`
`107°, 10836, 107°
`
`5. MgCl,
`
`108-8& 10° 8, 10°:
`
`
`
`
`
`
`6. MgCl, + D,O
`
`107%, 1088, 10° %°
`
`Type III Standard
`
`108 #2, 108% 10%
`
`*Outside limits
`
`.
`
`
`
`Mean Titre (95%
`Fiducial Limits)
`10°79
`(10°-°°-108-38)
`10°:3’
`(108 75-108 58)
`107:%
`(10°: 8-107-77)
`107°
`(10°-89-107-19)
`10°:%*
`(106: 8°-197:14)
`10° *°
`£1906 89-197-7%)
`
`
`(107-955-108 78)
`
`
`
`2. No Stabiliser
`+ D,O
`3. SPGA
`
`4. SPGA + D,O
`
`10°-45
`
`107°%
`
`10°°*°
`
`10°78
`
`10’ 06
`
`107°74
`
`5. MgCl,
`
`6. MgCl, + DO
`
`10°°*’
`
`107°
`
`10° -%
`
`10°:
`
`.
`
`
`
`
`10
`
`
`
`
`
`
`Day _7 at 4°C
`
`
`
`Sample
`Description
`1. No Stabiliser
`
`
`
`Total Virus Content
`(TCID,,/ml)
`10°-39
`10°-*°
`
`
`
`
`
`Type III Standard|10° 10°:?? 10°?
`
`20
`
`
`
`
`
`15
`
`Day 7 at 37°C
`
`
`
`Total Virus Content|Mean Titre (95%
`Sample
`(TCID.,/ml )
`Fiducial Limits)
`Description
`10?:8’
`107-8”
`102-7?
`1. No Stabiliser
`(102-59-102°9)
`
`2. No Stabiliser
`+ D,O
`3. SPGA
`
`4. SPGA + D,O
`
`25
`
`S. MgCl,
`
`6. MgCl,+D,0
`
`104°?
`
`10°:°°
`
`10° *
`
`105-8
`
`10°:*?
`
`104 -*
`
`10° °
`
`10° °°
`
`10°-%
`
`10° °°’
`
`10%°4?
`(104:73-104°%)
`10°:
`(10°-8®-10*-?°)
`108-6?
`(10°-44-108:78)
`
`10°91
`
`(10°:75-10° °)
`10° °°?
`
`(105:-9?-10°%-7°)
`Type III Standard|10° 108-73 1087?
`
`
`
`(107-°5-108-?8)
`
`
`
`
`
`
`
`Day 7 at 45°C
`
`
`Total Virus Content
`Mean Titre (95%
`(TCID.,/m1)
`Fiducial Limits)
`
`Sample
`Description
`
`1. No Stabiliser
`
`No viable poliovirus
`
`2. No Stabiliser
`+ D0
`
`No viable poliovirus
`
`
`
`
`
`
`
`
`
`
`
`
`
`
`10
`
`15
`
`20
`
`3. SPGA
`4. SPGA + D,0O
`5. MgCl,
`6. MgCl, + D,O
`
`No viable poliovirus
`10*°*?
`104°”
`10*:79
`(107*:52-10%4-97)
`No viable poliovirus Po
`10°-°*
`107-7
`1074
`"
`(103:7°-1093-5)
`
`Type III Standard
`
`10°"
`
`10°:}3
`
`10°13
`(107:-35-108-28)
`
`
`
`The results can be summarised as follows:
`
`
`
`
`
`
`
`
`
`Mean
`No Stabiliser
`SPGA
`MgCl,
`[+p,0|-p,0__| +p,0
`Titre
`
`
`
`
`
`
`
`
`
`10°:-19
`
`106 37
`
`107-°
`
`107:%
`
`10%: 36
`
`10°-98
`
`Day 7
`4a°c
`Day 7
`37°C
`10°°*?
`None
`107°”
`None
`None
`None
`Day 7
`25
`
`
`45°C detec-|detec-|}detec- detec-
`ted
`ted
`ted
`ted
`
`102:°77
`
`103 49
`
`10°:
`
`10°: 8
`
`103:%1
`
`10°:
`
`These results are represented graphically in Figures i,
`
`2
`
`30
`
`and 3 of the accompanying drawings. The initial titre of the
`
`Day 0 No Stabiliser formulation was low. This may have been due
`
`to the time that the virus stock stood at room temperature
`
`prior to blending. The target titre of 10°°°/ml was achieved at
`
`4°c after 7 days for the SPGA and MgCl, blends but not for the
`
`35
`
`blend with No Stabiliser. At
`
`the higher temperatures of 37°C
`
`
`
`
`
`+D,0
`
`6.42
`
`
`
`- 15 _
`
`and 45°C,
`
`the addition of D,O to the samples improved the
`
`stability of the virus. The loss in titre is expressed below as
`
`a logarithmic percentage of the initial antigen content:
`
`No Stabiliser
`
`SPGA
`
`MgCl,
`
`
`
`10
`
`15
`
`All percentages have been rounded up to the nearest whole
`
`number, hence the apparent 100% loss even where some antigen is
`
`detected. The figures in brackets were higher than the initial
`
`20
`
`(Day 0) titre. This apparent
`
`increase was due to the
`
`variability of the assay. The results emphasise the superiority
`
`of the formulations containing both SPGA and D,o.
`
`
`
`= 16 -
`
`CLAIMS
`
`A pharmaceutical composition which comprises a
`1.
`solution or suspension of a biological agent
`in a diluent
`comprising D,0, which solution or suspension contains
`sufficient amounts of a sugar, phosphate, glutamic acid ora
`physiologically acceptable salt thereof and, optionally, a non-
`toxic protein, a peptone or a mixture of two or more amino
`acids besides glutamic acid to enhance the stability of the
`
`agent.
`
`10
`
`15
`
`20
`
`A pharmaceutical composition according to claim 1,
`2.
`wherein the biological agent is a virus.
`3.
`A pharmaceutical composition according to claim 1,
`which is a suspension of a live attenuated poliovirus or an
`inactivated poliovirus of each of poliovirus types 1,
`2 and 3.
`4.
`A pharmaceutical composition according to any one
`of the preceding claims, wherein the solution or suspension
`contains from 0.5 to 3% by weight of a sugar,
`from 0.5 to 1.5%
`by weight of phosphate,
`from 0.2 to 1% by weight of glutamic
`acid or sodium glutamate and, optionally,
`from 0.5 to 3% by
`weight of the non-toxic protein, peptone or said mixture of
`
`25
`
`amino acids.
`A pharmaceutical composition according to any one
`5.
`of the preceding claims, wherein the solution or suspension
`contains (a) sucrose as the sugar,
`(b’) KH,PO, and {(b") K,HPO,
`as the phosphate,
`(c) L-glutamic acid or sodium glutamate and
`optionally (d) human serum albumin as the non-toxic protein,
`peptone or said mixture of amino acids in a weight ratio of
`(a): (b’): (b") : (c) soptionally (a) of about 1:0.40:0.36:0.37 when
`(c)
`is glutamic acid or 0.49 when (c)
`is sodium glutamate:0.46.
`
`
`
`_17 -
`
`6.
`
`A pharmaceutical composition according to any one
`
`of the preceding claims, wherein the solution or suspension
`
`comprises the biological agent and a D,0 SPG or SPGA solution
`
`as follows:
`
`SPG
`
`a solution in D,O of:
`
`1.09% w/v sucrose,
`
`0.43% w/v KH,PO,,
`
`0.39% w/v K,HPO,,
`
`and
`
`0.40% w/v L-glutamic acid or 0.53% w/v
`
`sodium glutamate;
`
`SPGA
`
`a solution in D,O of:
`
`1.09% w/v sucrose,
`
`0.43% w/v KH,PO,,
`
`0.39% w/v K,HPO,,
`
`0.40% w/v L-glutamic acid or 0.53% w/v
`
`sodium glutamate, and
`
`0.50% w/v human serum albumin.
`
`10
`
`15
`
`7.
`
`A process for the preparation of a pharmaceutical
`
`20
`
`composition as defined in claim 1, which process comprises
`
`dissolving or dispersing the biological agent
`
`in a D,0 solution
`
`of the said amounts of the sugar, phosphate, glutamic acid or
`
`salt thereof and, optionally, non-toxic protein, peptone or
`
`said mixture of amino acids.
`
`25
`
`8.
`
`A process according to claim 7, wherein the
`
`resulting solution or suspension is sterilised by passing
`
`through a sterilising filter and dispensed into unit- or multi -
`
`dose containers which are then sealed.
`
`9.
`
`A pharmaceutical composition which comprises a
`
`
`
`_ 18 -
`
`solution or suspension of a biological agent
`in a diluent
`comprising D,0, which solution or suspension is buffered to a
`pH at which the biological agent is stable by means of a
`phosphate buffer and contains sufficient amounts of a sugar,
`5 glutamic acid or a physiologically acceptable salt thereof and,
`optionally, a non-toxic protein, a peptone or a mixture of two
`or more amino acids besides glutamic acid to enhance the
`
`stability of the agent.
`10.
`Use of a D,O SPG or SPGA solution to stabilise a
`
`10 solution or suspension of a biological agent.
`
`
`
`
`
`
`
`A
`Patents Act 1977
`
`Application number
`Examiner’s report to the Comptroller under Section 17
`GB 9418609.5
`
`(The Search report)
`
`Search Examiner
`Relevant Technical Fields
`J F JENKINS
`
`
`Date of completion of Search
`18 DECEMBER 1995
`
`
`
`(ii) ONLINE: WPI, CAS - ONLINE
`
`(i) UK Cl (Ed.N)
`(ii) Int Cl (Ed.6)
`
`ASB (BAB, BLB, BNB)
`A61K 47/02, 47/16, 47/26, 47/36,
`47/18
`
`Databases (see below)
`(i) UK Patent Office collections of GB, EP, WO and US
`patent specifications.
`
`Documents considered relevant
`following a search in respect of
`Claims :-
`1 TO 10
`
`Categories of documents
`
`X:
`
`indicating lack of novelty or of
`Document
`inventive step.
`
` P:
`
`Document published on or afler the declared priority
`date but before
`the
`filing date of
`the present
`application.
`
`inventive step if
`indicating lack of
`Document
`Y:
`combined with one or more other documents of the=E: Patent document published on or after, but with
`
`same category.
`priority date earlier than, the filing date of the present
`application.
`
`A:
`
`indicating technological background
`Document
`and/or state of the art.
`
`&:
`
`Member of the same patent family, corresponding
`document.
`
`
`
`EP 0568726 A2 (RESEARCH FOUND. OSAKA UNIV.)|1, 2, 5 and 9
`see Reference Examples 5 and 6
`
`Relevant to
`claim(s)
`
`see Abstract
`
`EP 0332826 Al
`
`(TEVA PHARMACEUTICAL)
`see Claim 1
`
`WO 94/21298 Al
`
`(INSTITUT PASTEUR)
`see Example 4
`
`WPI Acc. No. 94 - 307582/38 & JP - 6234659
`(ZH-HANDAI BISEIBUTSUBYOK) 23 August 1994
`
`1-3
`
`1, 2, 5 and 9
`
`eS
`
`Databases:The UK Patent Office database comprises classified collections of GB, EP, WO and US patent specifications as outlined
`periodically in the Official Journal (Patents). The on-line databases considered for search are also listed periodically in the Official Journal
`(Patents).
`
`T8 - 25741
`
`Page 1 of 1
`
`
Accessing this document will incur an additional charge of $.
After purchase, you can access this document again without charge.
Accept $ Charge
This document could not be displayed.
We could not find this document within its docket. Please go back to the docket page and check the link. If that does not work, go back to the docket and refresh it to pull the newest information.
Your account does not support viewing this document.
You need a Paid Account to view this document. Click here to change your account type.
Your account does not support viewing this document.
Set your membership
status to view this document.
With a Docket Alarm membership, you'll
get a whole lot more, including:
- Up-to-date information for this case.
- Email alerts whenever there is an update.
- Full text search for other cases.
- Get email alerts whenever a new case matches your search.
One Moment Please
The filing “” is large (MB) and is being downloaded.
Please refresh this page in a few minutes to see if the filing has been downloaded. The filing will also be emailed to you when the download completes.
Your document is on its way!
If you do not receive the document in five minutes, contact support at support@docketalarm.com.
Sealed Document
We are unable to display this document, it may be under a court ordered seal.
If you have proper credentials to access the file, you may proceed directly to the court's system using your government issued username and password.
Access Government Site
