`
`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
`
`
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