United States Patent (19)
`Ondetti et al.
`
`3,937,819
`[11]
`(45) Feb. 10, 1976
`
`54 METHOD OF STABILIZING AN
`NJECTABLE COMPOSITION OF A
`CHOLECYSTOKININ ACTIVE
`OCTAPEPTIDE
`75) Inventors: Miguel Angel Ondetti, Princeton;
`Charles Rifkin, Edison, Bernard
`Rubin, Lawrence Township; Aaron
`L. Weiss, East Brunswick, all of N.J.
`73) Assignee: E. R. Squibb & Sons, Inc.,
`Princeton, N.J.
`July 22, 1974
`(22) Filed:
`21
`Appl. No.: 490,644
`
`OTHER PUBLICATIONS
`Sollmann, "A Manual of Pharmacology," Saunders
`Co., Phila., 1957, pp. 6, 7,549, 550.
`Kosower, "Introduction to Physical Organic Chemis
`try,” Wiley and Sons, New York, 1968, pp. 343-351.
`
`Primary Examiner-Lewis Gotts
`Assistant Examiner-Reginald J. Suyat
`Attorney, Agent, or Firm-Lawrence S. Levinson;
`. Merle J. Smith; Burton Rodney
`
`ABSTRACT
`57
`A stable composition of the sulfated octapeptide,
`
`52 U.S. C. ................................................ 424/177
`51
`Int. C.’.................. A61 K 37/00; CO7C 103/52
`58 Field of Search................... 424/1.77; 260/112.5
`
`(56)
`
`3,723,406
`3,734,946
`
`References Cited
`UNITED STATES PATENTS
`3/1973
`Ondetti et al.................... 26O11 12.5
`5/1973 Ondetti et al.................... 260f 1 12.5
`
`OH
`L-Asp-L-Tyr-L-Met-Gly-L-Trp-L-Met-L-Asp-L-Phe-NH
`having cholecystokinin activity, is obtained by lyophi
`lizing an aqueous solution of the octapeptide and
`NaCl.
`
`1 Claim, No Drawings
`
`
`
`
`MAIA Exhibit 1015
`MAIA V. BRACCO
`IPR PETITION
`
`

`

`1.
`METHOD OF STABILIZING AN NJECTABLE
`COMPOSITION OF A CHOLECYSTOKININ
`ACTIVE OCTAPEPTIDE
`
`BACKGROUND OF THE INVENTION
`The sulfated octapeptide
`
`OH
`L-Asp-L-Tyr-L-Met-Gly-L-Trp-L-Met-L-Asp-L-Phe-NH,
`
`has been found to possess cholecystokinin activity. As
`such it stimulates gall bladder contraction and is useful
`as a diagnostic aid in X-ray examination of the gall
`bladder in the same manner as cholecystokinin. For
`such purposes the sulfated octapeptide may be dis
`solved in water for injection to form an injectable
`which is administered either intravenously or subcuta
`neously to mammalian species, e.g., dogs or cats.
`OBJECTS OF THE INVENTION
`It is an object of the present invention to provide
`compositions of
`
`3,937,819
`2
`pH is adjusted with sufficient sodium hydroxide (as N
`solution) or hydrochloric acid (as 1 N solution), if
`necessary, to adjust the pH to from 5.50 to 6.50.The
`solution is brought to a volume of 1 liter by the addi
`tion of a sufficient quantity of water for injection.
`The foregoing solution is sterilized by filtration, asep
`tically filled into sterile vials, lyophilized, and sealed
`after filling the head space in the vial with sterile fil
`tered anhydrous nitrogen. The sealed vials are then
`stored at temperatures of 5°C or below.
`The lyophilized composition contains the sulfated
`octapeptide and sodium chloride. It has been found
`convenient to fill the vials before lyophilization with
`2.1 ml of a solution prepared as described above. The
`resulting vial after lyophilization then contains 5.25
`mcg of sulfated octapeptide and 45.0 mg of sodium
`chloride.
`The lyophilized material has excellent stability on
`storage and is readily reconstituted for injection by the
`addition of sterile water for injection. Preferably, the
`quantity of water for injection used for reconstitution is
`that amount which forms an isotonic solution.
`The following example illustrates the present inven
`tion without, however, limiting the same thereto. The
`sulfated octapeptide in each of the following examples
`S
`
`10
`
`15
`
`20
`
`25
`
`SOH
`L-Asp-L-Tyr-L-Met-Gly-L-Trp-L-Met-L-Asp-L-Phe-NH,
`
`SOH
`L-Asp-L-Tyr-L-Met-Gly-L-Trp-L-Met-L-Asp-L-Phe-NH2.
`
`30
`
`EXAMPLE
`The solution is prepared by adding 2500mcg of
`
`OH
`L-Asp-L-Tyr-L-Met-Gly-L-Trp-L-Met-L-Asp-L-Phe-NH,
`
`which are stable and retain the efficacy of the octapep
`tide during storage. Another object is to provide meth
`ods for preparing the stabilized compositions of the
`present invention. These and other objects of the pres
`35
`ent invention will be apparent from the following de
`scription.
`SUMMARY OF THE INVENTION
`A stable composition of the sulfated octapeptide
`
`40
`
`SOH
`L-Asp-L-Tyr-L-Met-Gly-L-Trp-L-Met-L-Asp-L-Phe-NH2
`
`is obtained by lyophilizing an aqueous solution of the
`sulfated octapeptide and sodium chloride.
`DETAILED DESCRIPTION
`A stable composition of the sulfated octapeptide,
`
`SOH
`L-Asp-L-Tyr-L-Met-Gly-L-Trp-L-Met-L-Asp-L-Phe-NH2
`
`45
`
`50
`
`and 21.43 g of sodium chloride to about 900 ml of
`water for injection, USP. If necessary, the pH is ad
`justed to between 5.50 and 6.50 with addition of slight
`amount of either a 1 N solution of sodium hydroxide or
`a 1 N solution of hydrochloric acid. The volume is then
`adjusted to 1.0 liter by addition of water for injection,
`USP.
`The solution is then filtered through a sterilizing
`membrane, and filled aseptically into 5 cc vials at 2.1
`ml/vial. The vials are stoppered with fluted stoppers in
`the raised position and frozen. The vials are then lyoph
`ilized for 24 hours at a temperature of -30°C, then for
`46 hours at a temperature of 25°C, and finally for one
`hour at a temperature of 37°C. The vials are then
`vented with dry sterile nitrogen and the stoppers placed
`in the closed position. The vials are then sealed and
`stored at a temperature of -20°C or lower.
`The lyophilized vial is reconstituted by addition of 5
`ml of sterile water for injection.
`What is claimed is:
`1. A method of enhancing the stability of the octa
`peptide
`
`55
`
`comprises a lyophilized powder of the sulfated octa
`peptide and sodium chloride.
`The preparation of the sulfated octapeptide perse is
`described in U.S. Pats. 3,723,406 and 3,734,946. The
`disclosure of these patents are incorporated herein by
`reference. As indicated therein the sulfated octapep
`tide possesses cholecystokinin activity, that is it stimu
`lates gallbladder contraction and so is useful as a diag
`nostic aid in X-ray examination of the gall bladder in
`the same manner as cholecystokinin.
`The compositions of the present invention are pre
`65
`pared from an aqueous solution of the sulfated octa
`peptide and sodium chloride. A liter of this solution
`contains 2500mcg and 21.43g of sodium chloride. The
`
`60
`
`SOH
`L-Asp-L-Tyr-L-Met-Gly-L-Trp-L-Met-L-Asp-L-Phe-NH.
`
`against degradation during storage, which comprises
`lyophilizing a solution containing per liter about 2500
`
`
`
`
`

`

`3,937,819
`4.
`3
`neg of said octapeptide and about 2143 g of Sodium
`sary, by addition of sodium hydroxide or hydrochloric
`chloride, and a sufficient quantity of water for injection
`to adjust the volume to about 1 liter, the pH of the
`acid.
`solution being adjusted to from 5.50 to 6.50, if neces-
`
`:k
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`sk
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`ck
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`k
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`:k
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`15
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`20
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`25
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`30
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`35
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`40
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