Umted States Patent
`
`[19]
`
`[in Patent Number:
`
`5 482 931
`9
`9
`
`Harris et al.
`
`[45] Date of Patent:
`
`Jan. 9, 1996
`
`USO05482931A
`
`[54] STABILIZED PHARMACEUTICAL PEPTIDE
`COMPOSITIONS
`
`[75]
`
`IIIVCIIKOISZ Alan Harris; Birgitta
`Tennhammar-El-rman, both of Malmo,
`Sweden
`
`[73] Assignee: Ferring AB, Malrno, Sweden
`
`[21] App1- N0-= 84,563
`[22] Filed:
`Jun. 29, 1993
`[51]
`Int. 0.5 .......................... A61K 38/00; C07K 14/00;
`C07K 7/16
`[52] U.S. Cl.
`................................... 514/15; 514/2; 514/12;
`530/315
`
`[56]
`
`Of Search .....................................
`.
`References cued
`U_s, PATENT DOCU1\/[ENTS
`
`12,
`
`_
`
`'
`
`2/1974 Kamber at 31'
`3,794,633
`.
`3,929,758 12/1975 Hughes et a1.
`_
`4,033,940
`7/1977 Hughes et a1.
`4,093,610
`6/1978 Abraham et al. _
`4,216’141
`3,1980 Rivier et al. _
`4,271,053
`5/1931 Kambe, et a1_ _
`4,351,754
`9/1932 Bin‘ _
`4,487,765 12/1984 de Wied .
`4,764,378
`8/1988 Keith et al.
`
`............................. 424/435
`
`4,985,242
`5,066,716
`5,124,315
`
`1/1991 Sekine et al.
`.
`11/1991 Robey et al.
`6/1992 Ceschel et al.
`
`............................. 514/12
`
`........................... 514/12
`
`FOREIGN PATENT DOCUMENTS
`
`.
`
`3/1993 European Pat. Off.
`03744
`9/1969 Gem“mY-
`1900367
`1/1985 Germany .
`2254043C2
`9/1990 G
`.
`3335086C2
`11/1986 U:1'1:dnI)£ingdom.
`0199922
`Primary Examiner—-Jill Warden
`‘,§l:rr"31'_1‘I‘,:’)‘I:‘(‘)‘L1d Calimafde Kalil &
`’
`’
`’
`
`’
`
`J“‘fl°“’°
`[57]
`
`ABSTRACT
`
`Disclosed is a Stabilized aqueous com osifion for adln:-u,1iS_
`.
`.
`.
`.
`b.
`1
`I? an
`.
`.d
`tration to a patient comprising a 10 ogic
`y active pepti e,
`a buifer, a quaternary amine-type preservative or disinfec-
`tarit, and an osmotic pressure-controlling agent, which com-
`position can be stored and used at room temperature. The
`buffer stabilizes the pH of the composition between about 4
`and 6. The preferred buifer contains citrate and/or. phos-
`phate, and the preferred preservative or disinfectant is ben-
`zalkonium chloride. The composition protects the peptide
`contained therein from adhering to container surfaces, par-
`ticularly in containers made of polymeric materials.
`
`-
`
`22 Claims, 1 Drawing Sheet
`
`   
`


`
`Lannett Holdings, Inc. LAN 1008
`
`

`
`U.S. Patent
`
`Jan. 9, 1996
`
`5,482,931
`
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`

`
`5,482,931
`
`1
`STABILIZED PHARMACEUTICAL PEPTIDE
`COMPOSITIONS
`
`FIELD OF THE INVENTION
`
`The present invention relates to stabilized aqueous phar-
`maceutical compositions for nasal, oral or parenteral admin-
`istration of small and medium-size peptides (up to about
`eicosapeptides), such as desmopressin (l—dearnino—8—D—
`arginine vasopressin; DDAVP).
`BACKGROUND
`
`10
`
`As used herein, the term “stabilized composition” refers
`to an aqueous solution for therapeutic use, containing at least
`one small or medium-size biologically active peptide. Such
`stabilization should allow the composition to be stored at
`room temperature for extended periods without
`loss in
`biological activity.
`A substantial number of biologically active peptides, their
`derivatives and analogs (in the following termed “peptides”)
`are known to be therapeutically useful. For various reasons
`they are often administered in form of aqueous compositons,
`that is, sterile aqueous solutions containing a known amount
`of peptide.
`The biological activity of the peptides to be administered
`is often extremely high. Thus, only very small amounts of
`peptide are needed for a single does. Such dilute aqueous
`peptide solutions in general are not stable at room tempera-
`ture for longer periods, even if kept in sealed containers. The
`therapeutically active peptide hormone analog desmo-
`spressin is such a peptide. Its aqueous solution has to be
`stored at a temperature not exceeding 8° C. Storage at higher
`temperaturs such as, for instance, room temperature, results
`in the degradation of desmopressin by hydrolytic and/or
`oxidative processes which are not blocked by the addition of
`a preservative, such as chlorobutanol (1,l,l-trichloro-2-
`methylpropan-2-ol). However, chlorobutanol effectively
`protects desmopressin against microbial attack.
`Another problem with dilute aqueous solutions of pep-
`tides is the adsorption of minute amounts of peptide to the
`walls of the container in which the solution is kept. Since
`such peptide solutions are usually very dilute, adsorption of
`even minor amounts may substantially reduce the amount of
`peptide available for administration.
`A particularly attractive way of administration of small
`and medium-size peptides in solution is via the nasal
`mucosa, either as drops or in spray form, which is even more
`convenient and more reproducible. Desmopressin,
`for
`instance, can be administered in an aqueous, 0.9 % sodium
`chloride solution (saline) by this route.
`Various kinds of intranasal spray delivery devices are
`known in the art. In general, peptides in an aqueous solution
`are administered by means of metered-dose spray pumps,
`such as those manufactured by Ing. Erich Pfeiifer KG,
`Radolfzell, Germany. An alternate route is via a graduated
`plastic tube of special design called a “rhinyle” which is
`partially filled with an aqueous solution containing a pep-
`tide. One end of the rhinyle is placed in the mouth and the
`other end is placed in the desired nostril, The solution is then
`delivered to the nostril by blowing.
`Peptides for nasal administration often have extremely
`high biological activity, and only a very small amount of
`peptide is needed in a single dose. However, the particular
`form of administration may require a minimum liquid vol-
`ume for good reproducibility. Thus, effective concentration
`ranges for nasally administered peptides are generally quite
`
`20
`
`25
`
`30
`
`35
`
`40
`
`45
`
`50
`
`55
`
`60
`
`65
`
`2
`low. For instance, a single desmopressin dose for nasal
`administration is typically between 10 pg to 40 pg, but may
`even be as little as 2.5 pg and as high as 300 pg. Typical dose
`volumes are from 100 pl to 400 pl (4X1OO pl). These doses
`are normally taken on a regular basis, such as at least once
`daily.
`
`‘ OBJECTS OF THE INVENTION
`
`Thus, it is an object of the present invention to overcome
`the aforementioned stability and storage problems associ-
`ated with known aqueous solutions of small and medium
`size peptides, particularly of aqueous solutions. containing
`desmopressin.
`Another object is to provide a stabilized aqueous solution
`containing a peptide for nasal, oral or parenteral adrninis—
`tration which can be conveniently stored at room tempera-
`ture for extended periods of time, for instance, one year,
`without risking partial or total degradation or microbial
`contamination of the peptide contained therein.
`A further object is to protect the peptide in solution from
`adhering to the walls of the container without using extra-
`neous additives specifically designed for that purpose.
`Yet another object is to provide an aqueous nasal or drop
`spray composition for the management of diseases and
`abnormal conditions which are mitigated by administration
`of small and medium sized, biologically active peptides.
`
`SUMMARY OF THE INVENTION
`
`invention is an aqueous composition for
`The present
`administration of small and medium-size peptides, particu-
`larly desmopressin, which can maintain stability over time
`and at room temperature, active biological ingredients car-
`ried therein such as a peptide, an analog of a peptide or
`mixtures of peptides and/or their analogs. The solution
`contains a bufier, a quaternary amine preservative or disin-
`feetant and an osmotic pressure-controlling agent.
`The quaternary amine preservative or disinfectant selec-
`tively used have, in addition to their namesake functions, the
`unexpected ability to prevent adsorption of small and
`medium size peptide components from adhering to container
`walls, particularly walls of containers made of polymeric
`materials.
`
`It is preferred for the peptide or peptide analog to be
`oxytocin or vasopressin, or their analogs and derivatives,
`such as particularly preferred, desmopressin (hereinafter
`also “DDAVP”).
`Also preferred are terlipressin (N-ot-triglycyl-8-1ysine)-
`vasopressin), atosiban ((Mpa1, D-Tyr(Et)2, Thr“, Orn8)-
`oxytocin), carbetocin ((l-desamino-1-monocarba-2(0-me-
`thyl)-tyrosine)-oxytocin) and triptorelin [D-Trpfi]-LI-IRH.
`It is preferred for the buifer to be capable of maintaining
`a pH of between 4.0 and 6.0. Especially preferred is a pH of
`about 5.0.
`
`In one embodiment, the buffer used is acetic acid/sodium
`acetate. It is preferred for the stabilized peptide solution
`according to the invention to contain citrate and/or phos-
`phate. Preferred buffer systems according to the invention
`are citric acid/disodium hydrogen phosphate, sodium dihy-
`drogen phosphate/disodium hydrogen phosphate, and citric
`acid/sodium citrate. Specifically preferred is a buffer com-
`prising: citratc-phosphate-sodium ions in a molar ratio of
`from about 1:323 to about 1:12.
`
`

`
`5,482,931
`
`3
`It is preferred for the quaternary amine preservative or
`disinfectant to be benzalkonium chloride, (NR1R2R3R4)“‘
`Cl"; where R‘, R2=methyl, R3=benzyl, R4=C3H17 to
`CISH37. The composition according to the invention pref-
`erably contains the quaternary amine preservative or disin-
`fectant in a concentration from about 0.05 to about 0.2 mg
`per ml. Particularly preferred is a concentration of about 0.1
`mg per ml.
`It is preferred for the osmotic pressure-controlling agent
`to be sodium chloride. The buffer components also contrib-
`ute substantially to osmotic pressure control.
`According to a preferred aspect of the invention the
`composition additionally contains at
`least one mucosal
`absorption enhancer such as bile salts, monolauryl ethers of
`macrogols, phospholipids, and fusidate derivatives.
`A preferred embodiment of the composition according to
`the invention contains from 0.025 mg to 1.5 mg of desmo-
`pressin acetate, from 1.35 to 1.75 mg of citric acid, from
`2.25 to 2.65 mg of disodium hydrogen phosphate, from 0.05
`to 0.20 mg benzalkonium chloride, and sodium chloride in
`an amount suflicient to provide the overall solution with an
`osmotic pressure comparable to that of human plasma.
`According to another preferred embodiment of the inven-
`tion, there is also disclosed the use of an aqueous spray
`composition for the management of diseases and abnormal
`conditions that can be treated by nasal administration of
`small and medium-size peptides.
`
`20
`
`25
`
`30
`
`BRIEF DESCRIPTION OF THE FIGURE
`
`FIG. 1 graphically illustrates the results of stability testing
`for the compositions prepared according to the present
`invention.
`
`35
`
`DETAILED DESCRIPTION
`
`The invention will now be explained in more detail by
`reference to the following experimental examples:
`
`40
`
`45
`
`Example 1
`
`4
`
`TABLE 1
`
`Stabilized desmopressin compositions (amounts per ml)
`A and B denote compositions according to the invention;
`C, D, E denote compositions prepared for comparison
`
`Compo-
`sition
`A
`
`B
`
`C
`
`D
`
`E
`
`NaCl BI-I
`mg mg (mmol)
`8.74 ACOH
`(mmol)
`2.96 ~ 103
`citric acid
`1.56
`citric acid
`1.97
`citric acid
`1.56
`
`6.29
`
`5.24
`
`6.30
`
`B‘Na*
`mg
`NaOAc
`0.58
`
`NaQHP04
`2.43
`Na2l-IP04
`1.83
`Na2HPO4
`2.43
`
`5.64
`
`citric acid
`1.97
`
`Na2HPO4
`1.83
`
`preservative
`mg
`benzalkonium
`chloride, 0.1
`
`benzalkoniurn
`chloride, 0.1
`benzyl alcohol
`10.0
`methyl p-hydroxy-
`benzoate*. 0.80
`propyl p-hydroxy—
`benzoate‘-‘*, 0.20
`chlorobutanol
`5.00
`
`*methy1 paraben; **propyl paraben
`
`Example 2
`
`STABILITY TESTING BY DETECTING
`PEPTIDE DEGRADATION
`
`The DDAVP-compositions prepared in Example 1 were
`stored in 10 ml glass vials (hydrolytic class 1, provided with
`Teflon® stoppers) in the dark at 65° C. for up for 13 weeks.
`Samples were taken after 1, 2, 3, 5, 7, 9, 11, and 13 weeks
`and analyzed for DDAVP by HPLC [Varian Star system;
`Lichrospher® PR-18 5 pm column (50><4 mm); gradient
`elution with various proportions of acetonitrile/0.0667M
`aqueous phosphate buffer pH
`The results are graphically depicted in FIG. 1 and dem-
`onstrate the superior stabilizing effect of the composition
`according to the invention. The experimental data contained
`in FIG. 1 were also used for calculation of first order rate
`constants shown in Table 2, below.
`
`PREPARATION OF TEST COMPOSITIONS
`
`50’
`
`desmopressin
`containing
`compositions
`test
`Five
`(DDAVP) acetate for nasal spray or drop compositions
`containing various preservatives were prepared, composi-
`tions A, B, C, D and E (see Table 1). Each test sample
`contained 0.089 mg DDAVP free base per ml, and Table 1
`denotes the type of bull’er used in each system. Millipore®-
`filtered water was used as solvent.
`
`Compositions A and B were prepared according to the
`present disclosure. Compositions C, D and E were prepared
`for comparative testing of other preservatives.
`The
`stability of
`the known, unbuffered Minirin®
`(DDAVP) spray containing NaCl and chlorobutanol has a
`useful shelf life of 3 years at refrigerated storage when kept
`in sealed glass containers. It is not stable at room tempera-
`ture when stored for longer periods of time. Note that
`composition E contains NaCl and chlorobutanol, but is a
`buifered solution.
`
`55
`
`60
`
`65
`
`TABLE 2
`
`First order rate constants for degradation of desmopressin
`
`Composition
`A
`B
`C
`D
`E
`
`k (s‘1)
`4.6 - 10'8
`8.0 - 10'°
`1.6 - 10-7
`8.8 - HTS
`~9 ~ 10"7
`
`correlation eoeflieient
`0.98
`0.98
`0.97
`0.96
`0.33
`
`Example 3
`
`CALCULATION OF USEFUL SHELF-LIFE
`
`From the slopes of the curves in FIG. 1 and from
`corresponding storage tests carried out at 37° C., 50° C., and
`60° C. Arrhenius activation energies (Ea) were obtained for
`compositions A, B, C and D. Composition E did not show
`Arrhenius-type behaviour since it was the least stable com-
`position by far.
`
`

`
`5,482,931
`
`5
`The storage time over which total DDAVP content of each
`composition was reduced by 10% (tgo) at 25° C. and 30° C.,
`“useful shelf life”, was calculated from E, which is tabulated
`below in Table 3.
`
`TABLE 3
`
`Useful shelf life in years (:90) for stabilized desmopressin
`compositions
`
`activation energy
`
`tgo
`
`10
`
`Composition
`A
`B
`C
`D
`
`(Ea, kl/mol)
`123.5
`115.1
`115.5
`102.8
`
`25” C.
`27.0
`12.9
`5.7
`7.4
`
`30° C.
`1 l .9
`6.0
`2.7
`3.7
`
`As Table 3 shows, desmopressin is preserved in compo-
`sitions A and B for extended periods of time at room
`temperature, thus demonstrating the ability of the present
`invention to be stored and used for extensive periods without
`refrigeration.
`
`Example 4
`
`COMPARISON OF INTRA-NASAL
`DESMOPRESSIN UPTAKE
`
`24 healthy fasting male subjects were given (randomized)
`desmopressin (20 pl ) intransally in spray form (200 pl ),
`using either compositon B or the commercially available
`unbufiered Minirin® formulation containing chlorobutanol
`as preservative. Blood samples were collected at intervals
`and desmopressin plasma levels monitored over a 12 h
`period by a desmopressin-specific RIA plasma assay (Lun-
`din, S. et al., Acta Endocrinologica (Copenhagen) 108
`(1985) 170-183). Essentially the same desmopressin plasma
`level profile was found for the two compositions.
`This is an unexpected result since H. A. Batts et al. (J.
`Pharm. Pharmacol. 1989, 156-159) found that chlorobu-
`tanol and benzalkonium chloride dilfered significantly in
`their eifect on the mucociliary transport rate in a frog palate
`model. The rate of mucociliary clearance affects the com-
`paratively slow intra-nasal uptake of peptides and other
`nasally administered biologically active compounds.
`
`Example 5
`
`ABSORPTION-BLOCKING EFFECT OF
`DESMOPRESSIN
`
`Sterile aqueou solutions of desmopressin marked with
`"SI (appr. 10,000 CPM/rnl) containing benzalkonium chlo-
`ride+saline, chlorobutanol+saline, or saline only, were all
`incubated in tubes of polystyrene, polypropene and glass for
`24 h at ambient temperature.
`In the solutions containing benzalkonium chloride and
`chlorobutanol, respectively, desmopressin showed insignifi-
`cant adsorption, whereas only about half of the amount of
`desmopressin in the preservative-free solution could be
`recovered from the plastic tubes.
`While the various features and embodiments of the
`present invention have been described herein, it is possible
`that one skilled in the art could modify the various aspects
`of the invention and obtain the same objectives. The present
`disclosure contemplates such modifications as being within
`its spirit and scope.
`
`20
`
`25
`
`30
`
`35
`
`40
`
`45
`
`50
`
`55
`
`60
`
`65
`
`6
`
`What is claimed is:
`1. A stable, aqueous composition for administration to a
`patient of at least one biologically active peptide, consisting
`essentially of:
`a) said biologically active peptide selected from the group
`consisting of oxytocin, vasopressin, an oxytocin ana-
`log, an oxytocin derivative, a vasopressin analog, and
`a vasopressin derivative;
`b) a bulfering agent;
`c) a quaternary amine preservative or disinfectant; and
`d) an osmotic pressure-controlling agent.
`2. The composition according to claim 1, wherein said
`peptide is selected from the group consisting of terlipressin,
`atosiban, carbetocin, and triptorelin.
`3. The composition according to claim 1, wherein said
`vasopressin analog is desmopressin.
`4. The composition according to claim 1, wherein said
`buffering agent is a buffer which maintains the pH of said
`composition between 4.0 and 6.0.
`5. The composition according to claim 4, wherein said
`buffer maintains said pH at about 5.0.
`6. The composition according to claim 1, wherein said
`buifering agent comprises a buffer selected from the group
`consisting of citrate, phosphate, and a mixture of citrate and
`phosphate.
`7. The composition according to claim 6, wherein said
`bufi‘er mixture of citrate and phosphate contains sodium ions
`such that the molar ratio of citrate, phosphate and sodium
`ions is from about 123:3 to about 1:122.
`8. The composition according to claim 1, wherein said
`quaternary amine preservative or disinfectant is benzalko-
`nium chloride having the following structure:
`
`(NR‘R’R3R“)*Cl‘
`
`where R1 and R2 are both methyl; R3 is benzyl; and R4 can
`be an alkyl group from CSHI-, to CRH37.
`9. The composition according to claim 1, wherein said
`administration is oral.
`10. The composition according to claim 1, wherein said
`administration is parenteral.
`11. An aqueous composition for nasal administration of a
`biologically active component, consisting essentially of:
`a) said biologically active component selected from the
`group consisting of a peptide selected from the group
`consisting of oxytocin, vasopressin, an oxytocin ana-
`log, an oxytocin derivative, a vasopressin analog, and
`a vasopressin derivative;
`b) a buffering agent;
`c) a quaternary amine preservative of disinfectant; and
`d) an osmotic pressure-controlling agent such the said
`composition is capable of maintaining said biologically
`active component in a functionally stable condition
`over extended periods and at room temperature.
`12. The composition of claim 11, wherein said vaso-
`pressin analog is desmopressin.
`13. The composition according to claim 11, wherein said
`buffering agent is a bufier which maintains the pH of said
`composition between 4.0 and 6.0.
`14. The composition according to claim 13, wherein said
`buffer maintains said pH at about 5.0.
`15. The composition of claim 11, wherein said buffering
`agent comprises a buifer selected from the group consisting
`of citrate, phosphate, and a mixture of citrate and phosphate.
`16. The composition of claim 15, wherein said buffer
`comprises a mixture of citrate and disodium hydrogen
`phosphate such that the molar ratio of citrate, phosphate and
`
`

`
`5,482,931
`
`7
`sodium ions is from about 1:3:3 to about 121:2.
`17. The composition according to claim 11, wherein said
`quaternary amine preservative or disinfectant is benzalko—
`nium chloride having the following structure:
`
`(NR‘R2R3R“)*Cl’
`
`where R1 and R2 are both methyl; R3 is benzyl; and R4 can
`be an alkyl group from C8H17 to C18H37.
`18. A stable aqueous composition for nasal application,
`comprising:
`a) from 0.025 mg to 1.5 mg of desmopressin acetate;
`b) from 1.35 mg to 1.75 mg of citric acid;
`c) from 2.25 mg to 2.65 mg of disodium hydrogen
`phosphate;
`d) from 0.05 mg to 0.20 mg of benzalkonium chloride;
`and
`
`8
`e) sodium chloride in an amount sufficient to provide said
`composition with an osmotic pressure comparable to
`that of human plasma.
`19. The stable, aqueous composition of claim 1, wherein
`said buifering agent can also function as said osmotic
`pressure-controlling agent.
`20. The stable, aqueous composition of claim 1, wherein
`said buffering agent in combination with sodium chloride
`can also function as said osmotic pressure-controlling agent.
`21. The aqueous composition of claim 11, wherein said
`buifering agent can also function as said osmotic pressure-
`controlling agent.
`22. The aqueous composition of claim 11, wherein said
`buifering agent in combination with sodium chloride can
`also function as said osmotic pressure-controlling agent.
`*
`*
`*
`*
`*
`
`

`
`UNITED STATES PATENT AND TRADEMARK OFFICE
`CERTIFICATE OF CORRECTION
`
`PATENT N0-
`DATOED
`
`:1
`;
`
`5,482,931
`January 9, 1996
`
`INVENTUNS) :
`
`Alan Harris and Birgitta Tennham'nar—Ekman
`
`It is certified that eiror appears in the above-identified patent and that said Letters Patent is hereby
`corrected as shown below:
`.
`
`In claim 11, column 6, line 50, delete "c-uf" and substitute
`
`therefor‘ —or——; and
`
`colurm 6, line 51 delete "the" ‘and
`
`A substitute therefor -—that-—.
`
`Signed and Sealed this
`
`Sixth Day of May, 1997
`
`mm
`
`BRUCE LEHMAN
`
`Arresting Oficer
`
`Commixsimzer of Palelzrx and Tradenzarkx