(12) INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT)
`
`(19) World Intellectual Property
`Organization
`International Bureau
`
`11111111111111111111 IIIII IIIII IIIIIIIII I II 11111111111111111111111111111111 11111111111111111
`
`(43) lntemationa1 Publication Date
`15 January 2004 (15.01.2-004)
`
`PCT
`
`(10) International Publication umber
`WO 2004/004781 Al
`
`(51) International Patent Classillcation7:
`38/37
`
`A61K47/00,
`
`STEVE S, John [GB/GB] ; J .D.S. Consulting, 18 Wood(cid:173)
`land Rd, Thornton Heath CR7 7LP (GB).
`
`(21) International Application Number:
`PCT /EP2003/007349
`
`(22) International Filing Date:
`
`8 July 2003 (08.07.2003)
`
`(25) Flllng Language:
`
`(26) Publication Lang11age:
`
`English
`
`English
`
`(74) Agent: GRUBB, Pltilip; Novartis AG, Corporate lntel (cid:173)
`]ectuel Property, CH-4002 Basel (CH).
`
`(81) Designated States (nationd.i) : AE, AG, AL, AM, AT, AU,
`AZ, BA, BB , BG, BR, BY, BZ, CA, CH, CN, CO, CR, CU,
`CZ, DE, DK, DM, DZ, EC, EE, ES, FI, GB, GD, GE, GH,
`HR, HU, ID, IL, IN, IS, JP, KE, KG KP, KR, KZ, LC, LK,
`LT, LU, LV, MA, MD. MK, MN, MX, NT, NO, NZ, OM,
`PG, PH, PL, PT, RO, RU, SC, SE, SG, SK, SY, TJ, TM,
`TN, TR, TT, UA, US, UZ, VC, VN, YU, ZA, ZW.
`
`(30) Priority Data:
`60/394,699
`60/394,6 12
`60/394,6 11
`
`9 Ju.ly 2002 (09.07.2002)
`9 July 2002 (09.07.2002)
`9 July 2002 (09.07.2002)
`
`(84) Designated States (regional): Eurasian patent (AM, AZ,
`BY, KG, KZ, MD, RU, TJ, TM), European patent (AT, BE,
`BG. CH, CY, CZ, DE, DK. EE, ES, FI, FR, GB, GR, HU.
`IE, IT, LU, MC, NL, PT, RO, SE, SI, SK, TR).
`
`us
`us
`us
`
`(71) Applicants (for all designa1ed Slates excepl US) : SAN(cid:173)
`DOZ G IBH [AT/AT]; Biochemiestrasse 10, A-6250
`Kundl (AT). GRANDIS BIOTECH GMBH [DE/DE);
`Grlinstras e 18, 79232 March-Hugstetten (DE).
`
`Published:
`with iniernaiional search report
`before the expiration of the time limit for amending the
`claimJ· cind to be republfahed in the ePenl of receipt of
`am1mdmenh·
`
`(72) Inventors; and
`(75) Inventors/Applicants {for US only): BETZ, Michael
`[DE/CH] ; Jligerstrasse I, CH-8200 Schaffhausen (CH).
`
`For two -letter codes and oiher abbre11iatio11.s, refer 10 the "Gu id(cid:173)
`ance Notes on Codes and Abbreviarions" appearing ar the begin(cid:173)
`ning of each regular issue of the PCT Gazette.
`
`--
`
`~
`QO
`
`;;;;;;;;;;; -iiiiii -
`t-"" 0
`= --------------------------------------
`
`:;;.; (54) Title: LIQUID FORMULATIONS WITII HIGH CO
`0
`ING 1,2-PROLPYLENE GLYCOL
`0
`~ (57) Abstract: The present invention relates to liquid formulations of human growth hormone (hGH, sornatropin) which are storage
`0 stab le, show reduced or no crystallization on storage and are suitable for administration to the human or ani mal body. More particu(cid:173)
`~ larly, the invention relates to liq uid formulations of human growth hormone which are stable and exhibit minimal or no crystallization
`;;;;-i,- when stored at least for a time at temperature above refrigerntion temperatures.
`
`OF HUMAN GROWTH HORMONE (hgh) COMPRIS-
`
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`

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`WO 2004/004781
`
`PCT/EP2003/007349
`
`LIQUID FORMULATIONS WITH HIGH CONCENTRATION OF HUMAN GROWTH HORMONE (HGH) COMPRI
`ING 1,2- PROPYLENE GLYCOL
`
`The present invention relates to liquid formulations of human growth hormone (hGH,
`
`somatropin) which are storage stable, show reduced or no crystallization on storage and are
`
`suitable for administration to the human or animal body. More particularly, the invention
`
`relates to liquid formulations of human growth hormone which are stable and exhibit minimal
`
`or no crystallization when stored at least for a time at temperatures above refrigeration
`
`temperatures.
`
`Native hGH is a single polypeptide chain protein consisting of 191 amino acids. The protein
`
`is internally cross-linked by two disulphide bridges and in monomeric form exhibits a
`
`molecular weight of about 22kDa.
`
`A major biological effect of hGH is to promote growth throughout a range of organs and
`
`tissues in the b.ody. hGH is secreted in a pulsatile manner from the pituitary gland throughout
`
`life. The major biological effect of hGH is to promote growth. hGH responsive organs or
`
`tissues include the liver, intestine, kidneys, muscles, connective tissue and the skeleton.
`
`hGH deficiency can occur in all age groups. The consequences of hGH deficiency include
`
`reduction in bone density, shortness in stature in children, reduction in lean body mass and
`
`extracellular volume and increase in cardiovascular risk factors. Replacement therapy with
`
`recombinant hGH has proven safe and effective in reversing these effects, but requires
`
`repeated injections at regular intervals
`
`For example, hypopituitary dwarfism is a condition which is readily treated by administering
`
`hGH to a subject suffering the condition. Prior to the production of large quantities of hGH
`
`by recombinant means only limited amounts of hGH could be prepared by laborious
`
`extraction of pituitary glands from human cadavers. This practice carried with it risks
`
`associated with infectious agents, eg the agent responsible for Creutzfeldt-Jakob disease
`
`(CJD), and that these agents might be passed to the patient receiving hGH. The isolation of
`
`the hGH gene and the construction of transformed host cells expressing recombinant hGH in
`
`cell culture has opened up not only a more reliable, safer and more cost effective treatment
`
`of hypopituitary dwarfism, but the possibility of using hGH for treatment of other diseases
`
`and conditions as well. Accordingly, in the context of the present invention, hGH preferably
`
`designates recombinant human growth hormone. However, it will readily appreciated that
`
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`
`PCT/EP2003/007349
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`also human growth hormone isolated from natural sources can in principle likewise be
`
`included in a pharmaceutical for!Tlulation of the present invention.
`
`A long appreciated problem with aqueous liquid formulations of pharmaceutical proteins, not
`
`just hGH, is that of instability during storage over a period of time. hGH in aqueous solution
`
`is known to undergo a variety of degradative changes. In common with most other proteins,
`
`Somatropin (recombinant human growth hormone, rhGH) has three main potential routes of
`
`degradation, namely hydrolysis leading to deamidation of free amide groups, oxidation of
`
`sulphur containing amino acids, and physical change of aggregation, where two or more
`
`hGH molecules physically stick together, for example, resulting in the formation of opaque
`
`insolubles. There is also the possibility of a clipping of the peptide backbone as a result of
`
`hydrolysis. Additionally, a major problem is crystallization of hGH.
`
`Early suggestions about how to solve the problems of instability noted above included freeze
`
`drying, but this of course meant that the resulting lyophilised product needed reconstitution
`
`immediately or shortly prior to administration. In the circumstances of routine self(cid:173)
`
`administration by a patient at home, this normally means that the patient has the task of
`
`reconstituting the lyophilised preparation into an aqueous solution. This is inconvenient for
`
`the patient and carries with it a risk of improper reconstitution due to lack of care, lack of
`
`attention to detail and instructions, or simply misunderstanding on the part of the patient.
`
`Freeze drying of formulations also suffers from the disadvantage of being costly and time
`
`consuming from a manufacturing perspective.
`
`Much effort is therefore expended in finding formulations which permit a simpler self(cid:173)
`
`administration of hGH by patients. These efforts are focused on ways of providing
`
`sufficiently stable aqueous liquid hGH formulations in a ready to use form. Such liquid
`
`dosage forms offer increased convenience and hence better compliance compared to
`
`lyophilized dosage forms which have to be reconstituted and filled into a pen cartridge via an
`
`additional device.
`
`However, care has to be taken that excipients which may be able to stabilize an aqueous
`
`formulation of hGH may carry some risk in administration to patients. Many compounds
`
`which may serve as stabilizers would not appear clinically acceptable and therefore would
`
`not enable a pharmaceutically acceptable formulation to be made. Furthermore,
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`PCT/EP2003/007349
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`-3-
`
`pharmaceutical regulatory requirements dictate that any unnecessary additives/ excipients,
`
`particularly synthetic additives I excipients, must be avoided in order to reduce risks to
`patients.
`
`Conveniently, aqueous pharmaceutical formulations of hGH should be offered as multi(cid:173)
`
`dosage formulations to the patient, who will administer such a formulation by means of an
`
`injector device. Such multi-dosage pharmaceutical formulations usually require an
`
`appropriate preservative to be present.
`
`Common liquid formulations of hGH are known to contain the drug at a low concentration,
`
`e.g. about 3.33 mg/ ml, which, however, upon administration may cause certain
`
`disadvantages for the patient.
`
`In particular, a patient has to receive a relatively large volume of such a low-concentration
`
`formulation of hGH per injection, which may cause discomfort or even pain. For example, for
`
`children suffering from growth hormone deficiency (GHD) hGH may have to be administered
`
`at a dosage of about 0.1 IU / kg bodyweight/ day. Accordingly, a patient having a
`
`bodyweight of 50 kg would have to receive about 5 IU hGH per day, which is contained in
`
`500 µI of a liquid formulation comprising about 3.33 mg/ ml hGH (1 IU hGH = 0.33 mg
`
`hGH). It will readily be appreciated that the application of a volume of less than 500 µJ would
`
`be highly desired.
`
`In the alternative, such a dosage could be administered in 2 or more injections of such a
`
`low-concentrated hGH formulation, each injection having a reduced volume. However, in
`
`terms of application safety, the use of more than one injection per dosage is not
`
`recommended.
`
`Furthermore, depending on the treatment schedule and dosage, a patient may have to use
`
`more than one single injection of such a low-concentration hGH formulation in order to be
`
`able to provide the prescribed amount of hGH. This may apply for example to patients
`
`having growth deficiency related to the Turner-Syndrome, who because of their increased
`
`body weight may be in need of a high amount of hGH. In many instances it will not be
`
`possible to deliver the required amount of hGH to such patients with a single injection having
`
`a reasonable volume of a such low-concentrated hGH formulation.
`
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`-4-
`
`Therefore, there is an ongoing need for a liquid pharmaceutical formulation containing hGH
`at a high concentration.
`
`In the course of the present invention it has been noticed that crystals tend to form in known
`
`aqueous, liquid growth hormone formulations if the concentration of hGH is adjusted to
`
`higher values, e.g. to 5 mg/ml hGH or more, in such formulations. This does not only apply
`
`just when such formulations are stored at refrigeration temperatures, but also when they are
`
`stored above refrigeration temperatures, at least for a time. The presence of crystals in
`
`liquid hGH formulations is highly undesirable because prior to administration such
`
`formulations need to be agitated or swirled and there may be instances when crystals are
`
`small or unobserved and the formulation is caused to be administered without dissolving the
`
`crystals sufficiently first. There is also the obvious disadvantage in terms of the visual
`
`appearance of hGH formulations when crystals have formed during storage.
`
`An object of the invention is therefore to provide a multi-dosage, aqueous liquid hGH
`
`formulation which is stable when stored for periods of time at refrigeration temperatures, e.g.
`
`for several months, or even for 1 or 2 years. Another object of the invention is to provide
`
`liquid hGH formulations which are stable when stored for at least a period of time above
`
`common refrigeration temperatures (e.g. above 2°c - 8°C} or even outside a refrigerator,
`
`e.g. for periods of several hours, days, or even weeks.
`
`In the context of the present application, "stable" mainly means that the problem of crystal
`
`formation is essentially avoided; preferably this problem is avoided completely. Accordingly,
`
`pharmaceutical formulation of the present invention exhibit minimal or no crystallization upon
`
`storage as decribed above.
`
`In addition to avoiding crystallization, a stable formulation should preferably show no or
`
`minimal aggregation of hGH upon storage. Likewise, a stable formulation preferably should
`
`not or only to a minimal extent undergo other degradation of hGH, e.g. by deamidation,
`
`oxidation and/or hydrolysis.
`
`In the context of the present invention, it has been developed that 1,2-propylene glycol to be
`
`used in such a multi-dosage liquid formulation containing a high concentration of hGH is a
`
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`-5-
`
`favourable parameter regarding stability. Furthermore, in the context of the present
`
`invention, it has been surprisingly established that a stable formulation can be composed of
`
`a smaller number of excipients than previously thought.
`
`Accordingly, an embodiment of the present invention relates to the use of 1,2-propylene
`
`glycol as a stabilizing agent in the preparation of a multi-dosage aqueous liquid
`
`pharmaceutical formulation comprising a high concentration of human growth hormone, as
`
`described herein.
`
`During the development of the present invention it has been shown that 1,2-propylene glycol
`
`is capable of providing stability to the pharmaceutical formulation and, simultaneously, it
`
`contributes to the desired tonicity of the formulation.
`
`In the context of the present invention, a liquid pharmaceutical formulation is a formulation
`
`provided in a ready-to-use form, i.e. it is not provided in a form to be reconstituted before
`
`administration, like e.g. a lyophilisate.
`
`The present invention therefore provides a multi-dosage liquid pharmaceutical formulation of
`
`human growth hormone consisting essentially of human growth hormone at a concentration
`
`of from about 5 mg/ml to about 100 mg/ml, 1,2-propylene glycol, an aqueous buff er, a non(cid:173)
`
`ionic surfactant and a preservative, said pharmaceutical formulation having a tonicity of from
`
`about 100 mosm/kg to about 500 mosm/kg and having a pH of from about 6.1 to about 6.3.
`
`Where necessary, additionally a tonicity-adjusting agent may be present in such a
`
`pharmaceutical formulation such that the tonicity is from about 100 to about 500 mosm/kg.
`
`Preferably, the pharmaceutical formulation of the present invention is isotonic.
`
`Accordingly, in a further embodiment thereof, there is provided a multi-dosage liquid
`
`pharmaceutical formulation of human growth hormone consisting essentially of human
`
`growth hormone at a concentration of from about 5 mg/ml to about 100 mg/ml, 1,2-propylene
`
`glycol, an aqueous buffer, a non-ionic surfactant and a preservative, said pharmaceutical
`
`formulation having a tonicity of from about 100 mosrn/kg to about 500 mosm/kg and having
`
`a pH of from about 6.2 to about 6.3, said pharmaceutical formulation additionally comprising
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`- 6 -
`
`a tonicity-adjusting agent such that the tonlcity of the pharmaceutical composition is from
`
`about 100 to about 500 mosm/kg.
`
`The presence of an additional tonicity-adjusting agent will be necessary if the further
`
`excipients of the formulation cannot contribute to the formulations' overall tonicity to such an
`
`extent that the desired-tonicity is achieved. In particular, depending on its concentration,
`
`1,2-propylene glycol is capable to provide a substantial part of the desired tonicity.
`
`Particularly pref erred are those pharmaceutical formulations according to the present
`
`invention, where 1,2-propylene glycol, together with the further excipents, is capable of
`
`providing the desired tonicity without the need of an additional tonicity-adjusting agent to be
`
`present, thereby keeping the overall number of excipients to be used to a minimum.
`
`In the context of the present invention, the term "consisting essentially ot· means that the
`pharmaceutical formulation of the present invention does not contain further excipients,
`
`besides the ones mentioned herein, which are capable to contribute a technological
`
`pharmaceutical function to the pharmaceutical formulation, e.g. in terms of stability, pH,
`
`tonicity, and the like. This does, however, not exclude the possibility that such a formulation
`
`may comprise one or more further auxiliary agents, which do not perform a technological
`
`pharmaceutical function in the formulation. Such auxiliary agents for example may be
`
`pharmaceutically acceptable dyes which will make the liquid formulation coloured. This may
`
`e.g. help in identifying the amount of liquid in a multi-dosage injection device or assist in
`
`easily identifying whether or not crystallization has occurred.
`
`Arising out of the present invention the inventors have perceived an advantage for patients,
`
`pharmacists and medical practitioners. Hitherto it has been necessary to ensure careful
`
`storage of growth hormone formulations at refrigeration temperatures (e.g. in the range of 2°
`
`to 8°C) in order to minimize crystallization. Prior to receipt of the growth hormone by
`
`patients the formulations can usually be reliably stored at refrigeration temperatures by
`
`manufactures and pharmacists. However, once received and stored by patients in domestic
`
`refrigerators there is much less reliability in terms of storage temperature. Temperatures in
`
`patients' domestic refrigerators may well be substantially above 2-8°C, e.g. be about 15°C,
`
`e.g. because of frequent opening. Moreover, devices containing the liquid formulation to be
`
`applied may stored outside the refrigerator, e.g. being forgotten on the kitchen bench after
`
`administration, thereby being exposed to room temperature (e.g. about 20°c to about 27°C,
`
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`-7-
`
`frequently about 25°C) for some time. Crystallization of hGH tends to occur more readily at
`
`temperatures greater than 8°C, i.e. above refrigeration temperatures, with known
`
`pharmaceutical formulations of hGH.
`
`The formulations of the present invention provide a greater resistance to crystallization if
`
`stored for a time above refrigeration temperatures. This therefore permits patients to be
`
`supplied with sufficient growth hormone to provide daily doses over longer periods of time
`
`than was hitherto recommendable or desirable. Whereas before, patients might have kept a
`
`small number of doses for use over a period of a week, with the formulations of the present
`
`invention patients may keep several weeks or even several months supply of growth
`
`hormone in domestic refrigerator with no or only minimal crystallization taking place. The
`
`frequency of prescription to patients can therefore be reduced significantly by the present
`
`invention.
`
`Accordingly, the pharmaceutical formulations of the present invention are stable, in particular
`
`substantially free of crystallization, on storage at temperatures from refrigeration
`
`temperatures to room temperature. In particular, such formulations are stable upon storage
`
`at temperatures from refrigeration temperatures to room temperature for at least 4 weeks or
`
`at least 1 month, preferably for at least 7 weeks, more preferably for at least 13 weeks. In a
`
`preferred embodiment thereof, such formulations are stable, in particular substantially free of
`
`crystallization, upon storage at temperatures between 2°c - 8°C for several months, e.g. for
`
`3 months, more preferably for at least 12 months, and most preferably for at least 18
`
`months. In a further preferred embodiment thereof, such formulations are stable, in particular
`
`substantially free of crystallization, at temperatures between 15°C and 25°C for at least 7
`
`weeks, more preferably for at least 13 weeks.
`
`In this context, it is to mention that prior to storage, hGH formulations may comprise about
`
`4% of "related proteins" being proteinaceous materials generated by degradative processes
`
`of deamidation and oxidation. Such "related proteins" are defined in the European
`
`Pharmacopoeia and measured by reversed phase HPLC. The inventors propose a maximum
`
`of 20% "related proteins" as a target at the end of the shelf life of the formulations.
`
`The degradation rate of hGH is not exactly linear and the rate of degradation increases with
`
`an increase in temperature. At 2° - 8°C formulations usually exhibit an increase in "related
`
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`- 8 -
`
`proteins" of about 0.8% per month. At 25°C this rises to about 13% per month, and at 40°C
`
`to about 70% per month. Storage at 25°C for 1 month is approximately equivalent to 17
`
`months storage at 2° - 8°C. Storage at 15°C for 1 month is approximately equivalent to 5
`
`months storage at 2° - 8°C. Continuous storage at a temperature in the range of about 25°
`
`to 40°C is the ref ore impractical.
`
`Although the formulations of the present invention off er good resistance to crystallization
`
`even up to 40°C, particularly up to 25°C, more particularly up to 15°C, the rapid formation of
`
`"related proteins" at these temperatures will usually place a more immediate limit on the
`
`potential shelf life of formulations.
`
`Rates of "related proteins" formation at different temperatures over time are readily
`
`measured by one of average skill and with this information the optimisation and maximum
`
`storage time/temperature patterns may be calculated without undue burden. In practice,
`
`formulations of the present invention can readily be subjected to a daily rise in temperature
`
`slightly above about 8°C due to the opening and closing of a refrigerator door or removal
`
`from a refrigerator for periods of an hour or so each day for the purposes administration
`
`without significant loss of shelf life. Advantageously, formulations of the present invention
`
`would not suffer adversely in terms of degradation or crystallization if left out of the
`
`refrigerator at room temperature for a day or so.
`
`Accordingly, the pharmaceutical formulations of the present invention may be kept at
`
`refrigeration temperature (e.g. in the range of 2° to 8°C) all the time in a stable condition.
`
`Furthermore, the pharmaceutical compositions show a sufficient stability when at least some
`
`of the overall storage time will be at a temperature above refrigeration temperatures,
`
`possibly up to about a week outside a refrigerator, possibly up to about a month or even
`
`longer outside a refrigerator.
`
`Accordingly, at least a part of the time that the formulation is stored may be at a storage
`
`temperature of at least 8°C, optionally a temperature in the range selected from 8° to 40°C,
`
`8° to 25°C or 8° to 15°C.
`
`In a preferred embodiment of the pharmaceutical formulations according to the present
`
`invention, the concentration of hGH in the formulation is from about 6 mg/ml to about 14
`
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`-9-
`
`mg/ml. In a particularly preferred embodiment thereof, the concentration of hGH in the
`
`formulation is about 6.67 mg/ml.
`
`In the development of the present invention it has surprisingly been established that
`
`1,2-propylene glycol is capable of providing sufficient stability to the formulations of the
`
`present invention which comprise such a high concentration of hGH. Preferably, the
`
`pharmaceutical formulations of the present invention comprise 1,2-propylene glycol at a
`
`concentration of from about 0.5 mg/ml to about 20 mg/ml, more preferably from about 5
`
`mg/ml to about 15 mg/ml, most preferably of from about 6 mg/ml to about 13 mg/ml.
`
`Pref erred embodiments relate to pharmaceutical formulations according to the present
`
`invention which contain about 9 mg/ml and 12,4 mg/ml 1,2-propylene glycol.
`
`The aqueous buffer present in the pharmaceutical formulation of the present invention can
`
`be any pharmaceutically acceptable buffer. In a preferred embodiment thereof, the aqueous
`
`buffer is selected from the group consisting of a phosphate buffer, a citrate buffer, an
`
`acetate buffer and a formate buffer, preferably a phosphate buffer, more preferably a sodium
`
`phosphate buffer. Usually, the aqueous buffer has a concentration of from about 5 mM to
`
`about 100 mM. In a preferred embodiment thereof, the aqueous buffer has a concentration
`
`of about 10 mM. In a particularly preferred embodiment thereof, the aqueous buffer is a
`
`phosphate buffer having a concentration of about 10 mM (the number 10 mM referring to the
`
`concentration of the phosphate ions). Most preferably the aqueous buffer is a sodium
`
`phosphate buffer having a concentration of about 10 mM. Likewise preferred is a 10 mM
`
`phosphate buffer, in particular a 10 mM sodium phosphate buffer.
`
`The non-ionic surfactant present in the pharmaceutical formulation of the present invention
`
`can be any non-ionic surfactant which is pharmaceutically acceptable. Preferably, the non(cid:173)
`
`ionic surfactant is selected from the group consisting of poloxamers, such as poloxamer 184
`
`or 188, and polysorbates, such as polysorbate 20 or 80, for example, and other
`
`ethylene/polypropylene block polymers. Preferably, the non-ionic surfactant is a poloxamer,
`
`in particular poloxamer 188. Amounts of the non-ionic surfactant used may be in the range
`
`from about 0.001 % (w/v) to about 10% (w/v), more preferably from about 0.005% (w/v) to
`
`about 5% (w/v), even more preferably from about 0.01% (w/v} to about 1% (w/v). In a
`
`preferred embodiment thereof, the non-ionic surfactant is present at a concentration of from
`
`about 0.05 mg/ml to about 4 mg/ml, preferably at a concentration of about 2 mg/ml. A
`
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`
`preferred embodiment of the present invention relates to a pharmaceutical formulation
`
`wherein the non-ionic surfactant is poloxamer 188 present at a concentration from about
`
`0.05 mg/ml to about 4 mg/ml, preferably of about 2 mg/ml.
`
`The preservative present in the pharmaceutical formulation of the present invention can be
`
`any pharmaceutically acceptable preservative. Preferably, the preservative is selected from
`
`the group consisting of benzyl alcohol, meta-cresol, methyl paraben, propyl paraben, phenol,
`
`benzalkonium chloride, benzethonium chloride, chlorobutanol, 2-phenoxyethanol, phenyl
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`mercuric nitrate and thimerosal. The concentration of the preservative will be readily
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`available to those skilled in the art in agreement with requirements of health authorities
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`regarding the safety of multi-dosage formulations. Accordingly, the concentration of the
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`preservative can be, for example, from about 1 mg/ml to about 30 mg/ml, depending on the
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`preservative actually used. More preferably, the preservative is benzyl alcohol. In a preferred
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`embodiment thereof, the pharmaceutical formulation according to the present invention
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`comprises benzyl alcohol as preservative being present at a concentration of from about 7
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`mg/ml to about 12 mg/ml, most preferably at a concentration of about 9 mg/ml.
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`If in the pharmaceutical formulation according to the present invention an additional tonicity(cid:173)
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`adjusting agent is present for adjusting the tonicity of the formulation to a desired value from
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`about 100 mosm/kg to about 500mosm/kg, such tonicity-adjusting agent can be any
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`pharmaceutical acceptable tonicity-adjusting agent. Preferably, such tonicity-adjusting agent
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`is selected from the group consisting of a sugar, a sugar alcohol, a further polyol, a neutral
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`salt and an amino acid. For example, a sugar can pe a monosaccharide or a disaccharide,
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`like e.g. lactose or sucrose. For example, a neutral salt can be an inorganic salt, an organic
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`salt, or a mixed salt, displaying an about neutral pH upon dissolution in water, like e.g.
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`sodium chloride or ammonium acetate. For example, an amino acid can be glycine, arginine
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`or histidine. In a preferred embodiment thereof, the tonicity adjusting agent is a sugar
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`alcohol, preferably mannitol. The tonicity-adjusting agent preferably is present at a
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`concentration up to 70 mg/ml, more preferably up to 50mg/ml, even more preferably up to 30
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`mg/ml. In a particularly preferred embodiment thereof, the additional tonicity-adjusting agent
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`is mannitol at a concentration of about 30 mg/ml.
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`The pharmaceutical formulations according to the present invention preferably may have a
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`tonicity from about 100 mosm/kg to about 500 mosm/kg, i.e. the tonicity of such formulations
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`can be from hypotonic up to hypertonic. In a preferred embodiment thereof, the
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`pharmaceutical formulations of the present invention have a tonicity from slightly hypotonic
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`to slightly hypertonic. Preferably and in accordance with common knowledge (see e.g.
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`Pharmaceutical Dosage Forms, Parenteral Medications, Volume 2; edited by: Kenneth E.
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`Avis; Herbert A. Lieberman; Leon Lachman; Marcel Dekker Inc., New York and Basel,
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`published: 04/01/1993, page 58-60), this corresponds to a tonicity from about 250 mosm/kg
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`to about 350 mosm/kg. In a particularly preferred embodiment thereof, the pharmaceutical
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`formulations of the present invention are isotonic. lsotonicity preferably corresponds to a
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`tonicity of from about 270 mosm/kg to about 328 mosm/kg. More preferably isotonicity
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`corresponds to a tonicity of about 286 mosm/kg.
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`In a preferred embodiment, the pH-value of the pharmaceutical formulation according to the
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`present invention is about 6.2. A skilled person would understand a pH of about 6.2 to be
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`from pH 6.15 to pH 6.25. Preferably, the pH is 6.2.
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`A particularly pref erred pharmaceutical formulation of the invention essentially consists of
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`6.67 mg/ml human growth hormone,
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`from 6 mg/ml to 15 mg/ml 1,2-propylene glycol,
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`1 0 mM sodium phosphate buffer,
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`2 mg/ml poloxamer 188,
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`where necessary mannitol at a concentration sufficient such that the formulation is
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`substantially isotonic,
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`and having a pH of 6.2.
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`In further aspect there is provided a pharmaceutical composition essentially consisting of
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`6.67 mg/ml human growth hormone,
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`6 mg/ml 1,2-propylene glycol,
`1 o mM sodium phosphate buffer,
`22.5 mg/ml mannitol,
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`2 mg/ml poloxamer 188,
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`and having a pH of 6.2.
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`A stiU further aspect of the present invention relates to a pharmaceutical composition
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`essentially consisting of
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`6.67 mg/ml human growth hormone,
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`9 mg/ml 1,2-propylene glycol,
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`1 O mM sodium phosphate buffer,
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`8. 1 mg/ml mannitol,
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`2 mg/ml poloxamer 188,
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`and having a pH of 6.2.
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`In a yet further aspect the present invention relates to a pharmaceutical composition
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`essentially consisting of
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`6.67 mg/ml human growth hormone,
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`12.4 mg/ml 1,2-propylene glycol,
`1 o mM sodium phosphate buff er,
`2 mg/ml poloxamer 188,
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`and having a pH of 6.2.
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`The crystallization which is minimized or avoided in formulations by the present invention
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`appears to be that of growth hormone. Preferably any crystallization in the liquid formulation
`is detected directly by eye, more preferably under the light microscope at Sx magnification,
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`even more preferably under the light microscope at 1 Ox magnification. Prior to observation
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`under the light microscope formulations may be filtered and the presence or absence of
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`crystals on the filter determined. When viewing under the light microscope the filter may
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`have a pore size of about 5µm.
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`A particularly preferred test for crystallization is to store the formulation in a sealed container
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`with no airspace for a time period at 152C or at 25°C in the absence of light and then
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`observe the presence or absence of crystals by eye.
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`Furthermore, the aqueous growth hormone formulations of the present invention are
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`preferably storage stable in the sense that there is no or minimal aggregation of growth
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`hormone during the period of storage. Also, there is prefera