`
`i11111 11111111 III 11111 11111!!!11411111111211121,1111111111111111111111111
`
`
`
`(12) United States Patent
`Brunner-Schwarz et al.
`
`(10) Patent No.:
`(45) Date of Patent:
`
`US 7,476,652 B2
`Jan. 13, 2009
`
`(54) ACIDIC INSULIN PREPARATIONS HAVING
`IMPROVED STABILITY
`
`(75)
`
`Inventors: Anette Brunner-Schwarz, Frankfurt
`(DE); Norbert Lill, Kronberg (DE)
`
`(73) Assignee: Sanofi-Aventis Deutschland GmbH,
`Frankfurt (DE)
`
`( * ) Notice:
`
`Subject to any disclaimer, the term of this
`patent is extended or adjusted under 35
`U.S.C. 154(b) by 40 days.
`
`(21) Appl. No.: 11/089,777
`
`(22) Filed:
`
`Mar. 25, 2005
`
`(65)
`
`Prior Publication Data
`
`US 2005/0171009 Al
`
`Aug. 4, 2005
`
`Related U.S. Application Data
`
`(63) Continuation of application No. 10/461,740, filed on
`Jun. 13, 2003, now abandoned.
`
`(60) Provisional application No. 60/409,338, filed on Sep.
`9, 2002.
`
`8/1996 Anderson, Jr. et al.
`5,547,929 A
`9/1996 Brems et al.
`5,559,094 A
`1/1997 Brange
`5,597,796 A
`3/1997 Wunderlich et al.
`5,614,219 A
`8/1997 Dorschug
`5,656,722 A
`9/1997 Obermeier et al.
`5,663,291 A
`12/1997 Bechgaard et al.
`5,693,608 A
`12/1997 Chance et al.
`5,700,662 A
`1/1998 Gertner et al.
`5,707,641 A
`7/1998 Clark et al.
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`9/1999 Kimer et al.
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`12/2002 Havelund et al.
`
`(Continued)
`
`514/4
`514/3
`
`514/3
`
`424/426
`
`(30)
`
`Foreign Application Priority Data
`
`FOREIGN PATENT DOCUMENTS
`
`Jun. 18, 2002
`
`(DE)
`
`
`
` 102 27 232
`
`(51) Int. Cl.
`A61K 38/28
` 514/3; 514/4
`(52) U.S. Cl.
` None
`(58) Field of Classification Search
`See application file for complete search history.
`
`(2006.01)
`
`(56)
`
`References Cited
`
`AU
`AU
`CA
`CA
`CA
`CA
`
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`U.S. PATENT DOCUMENTS
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`
`(Continued)
`
`514/4
`
`Primary Examiner Jeffrey E Russel
`(74) Attorney, Agent, or Firm Balaram Gupta
`
`(57)
`
`ABSTRACT
`
`The invention relates to a pharmaceutical formulation com-
`prising a polypeptide selected from the group consisting of
`insulin, an insulin metabolite, an insulin analog, an insulin
`derivative and combinations thereof; a surfactant or combi-
`nations of two or more surfactants; optionally a preservative
`or combinations of two or more preservatives; and optionally
`an isotonicizing agent, buffers or further excipients or com-
`binations thereof, the pharmaceutical formulation having a
`pH in the acidic range.
`
`25 Claims, No Drawings
`
`Mylan Ex.1092
`Mylan v. Sanofi - IPR2018-01676
`
`

`

`US 7,476,652 B2
`Page 2
`
`U.S. PATENT DOCUMENTS
`
`WO
`
`WO 03/035051
`
`5/2003
`
` 514/3
`
`5/2004
`6,734,162 B2
`11/2004
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`6/2005
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`6,960,561 B2 * 11/2005
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`Havelund
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`Boderke
`
`FOREIGN PATENT DOCUMENTS
`
`DE
`DE
`EP
`EP
`EP
`EP
`EP
`EP
`EP
`EP
`EP
`EP
`EP
`EP
`EP
`EP
`EP
`EP
`EP
`EP
`EP
`EP
`EP
`EP
`FR
`GB
`WO
`WO
`WO
`WO
`WO
`WO
`WO
`WO
`WO
`WO
`WO
`WO
`WO
`WO
`WO
`WO
`WO
`WO
`WO
`WO
`WO
`WO
`WO
`WO
`WO
`WO
`WO
`WO
`WO
`
`2 219 635
`3 240 177
`0 018 609
`0 046 979
`0 166 529
`0 180 920
`0 194 864
`0 200 383
`0 211 299
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`0 227 938
`0 229 956
`0 254 516
`0 305 760
`0 368 187
`0 383 472
`0419504
`0 600 372
`0214826
`0 668 292
`0678522
`0 837 072
`0375437
`1 172 114
`2456522
`1527605
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`WO 96/07399
`WO 96/11705
`WO 96/41606
`WO 97/01331
`WO 98/42749
`WO 98/56406
`WO 99/24071
`WO 00/23098
`WO 00/23099
`WO 00/29013
`WO 00/74736
`WO 01/00223
`WO 01/12155
`WO 01/21154
`WO 01/28555
`WO 01/37808
`WO 01/43762
`WO 01/52937
`WO 01/93837
`WO 02/064115
`WO 02/076495
`WO 03/035028
`
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`Whittingham, J. L., et al., Insulin at PH2: Structural Analysis of the
`Conditions Promoting Insulin Fibre Formation, J. Mol. Biol., (2002),
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`Dixon et al., "Regeneration of Insulin Activity From the Separated
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`No. 1, 1989, pp. 148-173.
`Markussen et al., "Soluble, prolonged-acting insulin derivatives. I.
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`Markussen et al., "Soluble, prolonged-acting insulin derivatives. II.
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`Markussen et al., "Soluble, prolonged-acting insulin derivatives. III.
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`Muller et al., "Insulin Signaling in the Yeast Saccharomyces
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`
`Mylan Ex.1092
`Mylan v. Sanofi - IPR2018-01676
`
`

`

`US 7,476,652 B2
`Page 3
`
`Pillion et al., "Dodecylmaltoside-mediated Nasal and Ocular
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`Schartz et al., "A superactive insulin: [B10-Aspartic acid]insulin(hu-
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`
`Hinds et al., "Synthesis and Characterization of Poly(ethylene-gly-
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`Brange et al. "Neutral insulin solutions physically stabilized by addi-
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`
`* cited by examiner
`
`Mylan Ex.1092
`Mylan v. Sanofi - IPR2018-01676
`
`

`

`1
`ACIDIC INSULIN PREPARATIONS HAVING
`IMPROVED STABILITY
`
`This application is entitled to the benefit of U.S. Provi-
`sional Application 60/409,338, filed Sep. 9, 2002, and Federal
`Republic of Germany Application 10227232.8-41, filed Jun.
`18, 2002.
`
`SUMMARY OF THE INVENTION
`
`The invention relates to a pharmaceutical formulation
`comprising a polypeptide selected from the group consisting
`of insulin, an insulin metabolite, an insulin analog, an insulin
`derivative or combinations thereof; a surfactant or combina-
`tions of two or more surfactants; optionally a preservative or
`combinations of two or more preservatives; and optionally an
`isotonicizing agent, buffers or further excipients or combina-
`tions thereof, the pharmaceutical formulation having a pH in
`the acidic range. These formulations can be employed for the
`treatment of diabetes, and are particularly suitable for prepa-
`rations in which a high stability to thermal and/or physico-
`mechanical stress is necessary. The invention likewise relates
`to parenteral preparations which contain such formulations
`and can be used in diabetes and to methods for producing the
`preparations and for improving the stability of insulin prepa-
`rations.
`
`BACKGROAND OF THE INVENTION
`
`Worldwide, approximately 120 million people suffer from
`diabetes mellitus. Among these, approximately 12 million are
`type I diabetics, for whom the substitution of the lacking
`endocrine insulin secretion is the only currently possible
`therapy. The affected persons are dependent lifelong on insu-
`lin injections, as a rule a number of times daily. In contrast to
`type I diabetes, there is not basically a deficiency of insulin in
`type II diabetes, but in a large number of cases, especially in
`the advanced stage, treatment with insulin, optionally in com-
`bination with an oral antidiabetic, is regarded as the most
`favorable form of therapy.
`In the healthy person, the release of insulin by the pancreas
`is strictly coupled to the concentration of blood glucose.
`Elevated blood glucose levels, such as occur after meals, are
`rapidly compensated by a corresponding increase in insulin
`secretion. In the fasting state, the plasma insulin level falls to
`a basal value which is adequate to guarantee a continuous
`supply of insulin-sensitive organs and tissue with glucose and
`to keep hepatic glucose production low at night. The replace-
`ment of endogenous insulin secretion by exogenous, mostly
`subcutaneous administration of insulin, as a rule does not
`approximate the quality of the physiological regulation of the
`blood glucose described above. Often, deviations of blood
`glucose upward or downward occur, which in their severest
`forms can be life-threatening. In addition, however, blood
`glucose levels which are increased for years without initial
`symptoms are a considerable health risk. The large-scale
`DCCT study in the USA (The Diabetes Control and Compli-
`cations Trial Research Group (1993) N. Engl. J. Med. 329,
`977-986) demonstrated clearly that chronically elevated
`blood glucose levels are essentially responsible for the devel-
`opment of diabetic late damage. Diabetic late damage is
`microvascular and macrovascular damage which is mani-
`fested, ander certain circumstances, as retinopathy, nephr-
`opathy or neuropathy and leads to loss of sight, kidney failure
`and the loss of extremities and is moreover accompanied by
`an increased risk of cardiovascular diseases. In view of this,
`an improved therapy of diabetes should be aimed at keeping
`
`US 7,476,652 B2
`
`5
`
`30
`
`2
`the blood glucose as closely as possible in the physiological
`range. According to the concept of intensified insulin therapy,
`this should be achieved by repeated daily injections of rapid-
`and slow-acting insulin preparations. Rapid-acting formula-
`tions are given at meals in order to level out the postprandial
`increase in the blood glucose. Slow-acting basal insulins
`should ensure the basic supply with insulin, in particular
`during the night, without leading to hypoglycemia.
`Insulin is a polypeptide of 51 amino acids, which are
`10 divided into 2 amino acid chains: the A chain having 21 amino
`acids and the B chain having 30 amino acids. The chains are
`connected to one another by means of 2 disulfide bridges.
`Insulin preparations have been employed for diabetes therapy
`for many years. Not only are naturally occurring insulins
`15 used, but recently also insulin derivatives and analogs.
`Insulin analogs are analogs of naturally occurring insulins,
`namely human insulin or animal insulins, which differ by
`substitution of at least one naturally occurring amino acid
`residue with other amino acids and/or addition/removal of at
`20 least one amino acid residue from the corresponding, other-
`wise identical, naturally occurring insulin. The amino acids
`can in this case also be those which do not occur naturally.
`Insulin derivatives are derivatives of naturally occurring
`insulin or an insulin analog which are obtained by chemical
`25 modification. This chemical modification can consist, for
`example, of the addition of one or more specific chemical
`groups to one or more amino acids. As a rule, insulin deriva-
`tives and insulin analogs have a somewhat modified action
`compared with human insulin.
`Insulin analogs having an accelerated onset of action are
`described in EP 0 214 826, EP 0 375 437 and EP 0 678 522.
`EP 0 124 826 relates, inter alia, to substitutions of B27 and
`B28. EP 0 678 522 describes insulin analogs which in posi-
`tion B29 have various amino acids, preferably proline, but not
`35 glutamic acid. EP 0 375 437 includes insulin analogs with
`lysine or arginine in B28, which can optionally be addition-
`ally modified in B3 and/or A21.
`In EP 0 419 504, insulin analogs are disclosed which are
`protected against chemical modifications, in which aspar-
`40 agine in B3 and at least one further amino acid in the positions
`A5, A15, A18 or A21 are modified.
`In WO 92/00321, insulin analogs are described in which at
`least one amino acid of the positions B1 -B6 is replaced by
`lysine or arginine. According to WO 92/00321, insulins of
`45 this type have a prolonged action. The insulin analogs
`described in EP-A 0 368 187 also have a delayed action.
`The insulin preparations of naturally occurring insulins on
`the market for insulin substitution differ in the origin of the
`insulin (e.g. bovine, porcine, human insulin), and also the
`so composition, whereby the profile of action (onset of action
`and duration of action) can be influenced. By combination of
`various insulin preparations, very different profiles of action
`can be obtained and blood sugar values which are as physi-
`ological as possible can be established. Recombinant DNA
`55 technology today makes possible the preparation of such
`modified insulins. These include insulin glargine (Gly(A21)-
`Arg(B31)-Arg(B32)-human insulin) with a prolonged dura-
`tion of action. Insulin glargine is injected as an acidic, clear
`solution and precipitates on account of its solution properties
`60 in the physiological pH range of the subcutaneous tissue as a
`stable hexamer associate. Insulin glargine is injected once
`daily and is distinguished compared with other long-acting
`insulins by its flat serum profile and the reduction of the
`danger of nightly hypoglycemia associated therewith (Schu-
`65 bert-Zsilavecz et al., 2: 125-130(2001)).
`The specific preparation of insulin glargine, which leads to
`the prolonged duration of action, is characterized, in contrast
`
`Mylan Ex.1092
`Mylan v. Sanofi - IPR2018-01676
`
`

`

`US 7,476,652 B2
`
`3
`to previously described preparations, by a clear solution hav-
`ing an acidic pH. Especially at acidic pH, insulins, however,
`show a decreased stability and an increased proneness to
`aggregation on thermal and physicomechanical stress, which
`can make itself felt in the form of turbidity and precipitation
`(particle formation) (Brange et al., J. Ph. Sci 86:517-525
`(1997)).
`The proneness to aggregation can additionally be pro-
`moted by hydrophobic surfaces which are in contact with the
`solution (Sluzky et al., Proc. Natl. Acad. Sci. 88:9377-9381
`(1991). Surfaces which can be considered as hydrophobic are
`the glass vessels of the preparations, the stopper material of
`the sealing caps or the boundary surface of the solution with
`the air supernatant. In addition, very fine silicone oil droplets
`can function as additional hydrophobic aggregation nuclei in
`the taking of the daily insulin dose by means of customary,
`siliconized insulin syringes and accelerate the process.
`WO 01/43762 describes aqueous, parenteral pharmaceuti-
`cal preparations comprising a polypeptide and glycerol, in
`which the stabilization of the preparation is to be achieved by
`purifying off destabilizing constituents of the glycerol.
`WO 00/23098 describes insulin preparations stabilized
`using polysorbate 20 or poloxamer 188 for pulmonary admin-
`istration, but does not describe the stabilization in an acidic
`solution against aggregation nuclei.
`Published International patent application WO 02/076495
`describes zinc-free and low-zinc insulin preparations having
`improved stability at room and body temperature and to
`mechanical stress by the addition of surfactants, but does not
`describe the stabilization of acidic insulin preparations
`against hydrophobic aggregation nuclei.
`The present invention was thus based on the object of
`finding preparations for acid-soluble insulins containing sur-
`factants, which are distinguished by a high long-term stability
`to stress due to temperature or physicomechanical stressing
`and tolerate a high stress with hydrophobic aggregation
`nuclei.
`
`DETAILED DESCRIPTION OF THE INVENTION
`
`It has now surprisingly been found that the addition of
`surfactants can greatly increase the stability of acidic insulin
`preparations and thus preparations can be produced which
`guarantee superior stability to hydrophobic aggregation
`nuclei for several months ander temperature stress.
`The pharmaceutical preparations of the present invention
`contain 60-6000 nmol/ml, preferably 240-3000 nmol/ml, of
`an insulin, an insulin metabolite, an insulin analog or an
`insulin derivative.
`The surfactants which can be used are, inter alia, nonionic
`surfactants. In particular, pharmaceutically customary sur-
`factants are preferred, such as, for example: partial and fatty
`acid esters and ethers of polyhydric alcohols such as of glyc-
`erol, sorbitol and the like (Span®, Tween®, in particular
`Tween® 20 and Tween® 80, Myrj®, Brij®), Cremophor® or
`poloxamers. The surfactants are present in the pharmaceuti-
`cal composition in a concentration of 5-200 µg/ml, preferably
`of 5-120 µg/ml and particularly preferably of 20-75 µg/ml.
`The preparation can additionally optionally contain preser-
`vatives (e.g. phenol, cresol, parabens), isotonicizing agents
`(e.g. mannitol, sorbitol, lactose, dextrose, trehalose, sodium
`chloride, glycerol), buffer substances, salts, acids and alkalis
`and also further excipients. These substances can in each case
`be present individually or alternatively as mixtures.
`Glycerol, dextrose, lactose, sorbitol and mannitol are cus-
`tomarily present in the pharmaceutical preparation in a con-
`centration of 100-250 mM, NaCl in a concentration of up to
`
`4
`150 mM. Buffer substances, such as, for example, phosphate,
`acetate, citrate, arginine, glycylglycine or TRIS (i.e. 2-amino-
`2-hydroxymethy1-1,3 -propanediol) buffer and corresponding
`salts, are present in a concentration of 5-250 mM, preferably
`5 10-100 mM. Further excipients can be, inter alia, salts or
`arginine.
`The invention therefore relates to a pharmaceutical formu-
`lation comprising a polypeptide selected from the group con-
`sisting of insulin, an insulin analog, an insulin derivative, an
`0 active insulin metabolite and combinations thereof; a surfac-
`tant or combinations of two or more surfactants; optionally a
`preservative or combinations of two or more preservatives;
`and optionally an isotonicizing agent, buffer substances and/
`or further excipients or combinations thereof, the pharmaceu-
`15 tical formulation being a clear solution which has a pH in the
`acidic range (pH 1-6.8), preferably pH 3.5-6.8, very particu-
`larly preferably 3.5-4.5.
`Preferred pharmaceutical formulations of the present
`invention are those wherein the surfactant is selected from the
`20 group consisting of partial and fatty acid esters and ethers of
`polyhydric alcohols such as of glycerol and sorbitol, and
`polyols; the partial and fatty acid esters and ethers of glycerol
`and sorbitol being selected from the group consisting of
`Span®, Tween®, Myrj ®, Brij®, Cremophor®; the polyols
`25 being selected from the group consisting of polypropylene
`glycols, polyethylene glycols, poloxamers, Pluronics®, and
`Tetronics®; the preservative being selected from the group
`consisting of phenol, cresol, and parabens; the isotonicizing
`agent being selected from the group consisting of mannitol,
`30 sorbitol, sodium chloride, and glycerol; the excipients being
`selected from the group consisting of buffer substances,
`acids, and alkalis; the insulin analog being selected from the
`group consisting of Gly(A21)-Arg(B31)-Arg(B32)-human
`insulin; Lys(B3)-Glu(B29)-human insulin; Lys'ProB29
`35 human insulin, B28 Asp-human insulin, human insulin in
`which proline in position B28 has been substituted by Asp,
`Lys, Leu, Val or Ala and where in position B29 Lys can be
`substituted by Pro; AlaB26-human insulin; des(B28-B30)-
`human insulin; des(B27)-human insulin and des(B30)-hu-
`40 man insulin; the insulin derivative being selected from the
`group consisting of B29-N-myristoyl-des(B30) human insu-
`lin, B29-N-palmitoyl-des(B30) human insulin, B29-N-
`myristoyl human insulin, B29-N-palmitoyl human insulin,
`B28-N-myristoyl LysB28ProB29 human insulin, B28-N-
`45 palmitoyl-Ly'ProB29 human insulin, B30-N-myristoyl-
`Thr829LysB3°
`human
`insulin,
`B30-N-palmitoyl-
`ThrB29LysB3° human
`insulin, B29-N-(N-palmitoyl-y-
`glutamy1)-des (B30) human insulin, B29-N-(N-lithocholyl-y-
`glutamy1)-des (B30)
`human
`insulin,
`B29-N-(w-
`50 carboxyheptadecanoy1)-des (B30) human insulin and B29-N-
`(w-carboxyheptadecanoyl) human insulin.
`A further subject of the invention is a pharmaceutical for-
`mulation such as described above, in which the insulin, the
`insulin analog, the active insulin metabolite and/or the insulin
`55 derivative is present in a concentration of 60-6000 nmol/ml,
`preferably in a concentration of 240-3000 nmol/ml (this cor-
`responds approximately to a concentration of 1.4-35 mg/ml
`or 40-500 units/ml);
`
`60 in which the surfactant is present in a concentration of 5-200
`µg/ml, preferably of 5-120 µg/ml and particularly preferably
`of 20-75 µg/ml.
`A further subject of the invention is a pharmaceutical for-
`mulation such as mentioned above, in which glycerol and/or
`65 mannitol is present in a concentration of 100-250 mM, and/or
`NaCl is preferably present in a concentration of up to 150
`mM.
`
`Mylan Ex.1092
`Mylan v. Sanofi - IPR2018-01676
`
`

`

`5
`A further subject of the invention is a pharmaceutical for-
`mulation such as mentioned above, in which a buffer sub-
`stance is present in a concentration of 5-250 mM.
`A further subject of the invention is a pharmaceutical insu-
`lin formulation which contains further additives such as, for
`example, salts which delay the release of insulin. Mixtures of
`such delayed-release insulins with formulations described
`above are included therein.
`A further subject of the invention is a method for the
`production of such pharmaceutical formulations. Likewise, a
`further subject of the invention is the use of such formulations
`for the treatment of diabetes mellitus.
`A further subject of the invention is the use or the addition
`of surfactants as stabilizer during the process for the produc-
`tion of insulin, insulin analogs or insulin derivatives or their
`preparations.
`
`EXAMPLES
`
`The following examples illustrate, but by no means limit,
`the present invention.
`Comparison investigations: Different preparations con-
`taining the insulin analog insulin glargine (Gly(A21),Arg
`(B31),Arg(B32)-human insulin) are prepared. To this end,
`insulin glargine is suspended in one part of water for injec-
`tion, dissolved at pH 3-4, the other constituents are added, the
`pH is adjusted to 4.0+/-0.2 using hydrochloric acid/NaOH
`and the mixture is made up to the final volume. The concen-
`tration of insulin glargine in each of the experiments
`described below is 3.6378 mg/ml (corresponds to 100 units/
`ml). A second preparation is produced identically, but a spe-
`cific amount of a surfactant is additionally added. The solu-
`tions are filled into 10 ml glass vessels (vials) and fitted with
`crimp caps. These vessels are now exposed to simulated in use
`or physicomechanical stress conditions:
`1. In use test: The vessels are sorted into boxes with turned-up
`lids and stored during the investigation period of 28 days at
`+25° C. and controlled room humidity with exclusion of
`light. To simulate taking by the patient, once daily about 5
`IU of the solutions are withdrawn using a customary insu-
`lin syringe and discarded. At the beginning and end of the
`working week this procedure is carried out twice in order to
`to simulate taking at the weekend. Before each withdrawal,
`visual assessment of the solution in the vessels for turbidity
`and/or particle formation is carried out.
`2. Shaking test: The vessels are placed in a box with a turned-
`up lid lying on a laboratory shaker having an incubator and
`thermostat and shaken at 25° C. with 90 movements/min
`parallel to the horizontal movement for a period of time of
`10 days. After defined times, the turbidity value of the
`samples is determined by means of a laboratory turbidity
`photometer (nephelometer) in formaldazine nephelomet-
`ric units (formaldazine nephelometric unit—FNU). The tur-
`bidity value corresponds to the intensity of the scattered
`radiation of the light incident on suspended particles in the
`sample.
`
`Example 1
`
`Stabilization of the in Use Period of Insulin Glargine
`Using Polysorbate 20 (Tween® 20)
`
`a) The solution is sterile-filtered through a combination of 0.2
`nm and 0.1 nm filters. It is then poured into 10 ml injection
`vials and sealed using crimp caps having an inserted seal-
`ing disk.
`
`US 7,476,652 B2
`
`6
`b) A comparison solution is prepared identically, but first a
`suitable amount of surfactant (10-30 ppm of polysorbate
`20) is suspended in water for injection. The samples are
`stored at +5° C., 25° C. and 37° C. fora fixed period of time.
`10 samples in each case are then subjected to an in use test.
`The results are shown in the table below.
`
`Storage for 3 Months at 5° C.
`
`5
`
`10
`
`Number of vials with
`particle formation after
`
`Test sample
`
`7 days
`
`14 days 21 days 28 days
`
`15
`
`Insulin glargine
`Insulin glargine + 0.010 mg/ml
`of polysorbate 20
`Insulin glargine + 0.015 mg/ml
`of polysorbate 20
`Insulin glargine + 0.020 mg/ml
`20 of polysorbate 20
`Insulin glargine + 0.030 mg/ml
`of polysorbate 20
`
`7
`0
`
`10
`0
`
`10
`0
`
`10
`0
`
`0
`
`0
`
`0
`
`0
`
`0
`
`storage for 6 months at 5° C.
`
`25
`
`30 Test sample
`
`7 days
`
`14 days 21 days 28 days
`
`Number of vials with
`particle formation after
`
`Insulin glargine
`Insulin glargine + 0.010 mg/ml
`of polysorbate 20
`Insulin glargine + 0.015 mg/ml
`of polysorbate 20
`Insulin glargine + 0.020 mg/ml
`of polysorbate 20
`Insulin glargine + 0.030 mg/ml
`of polysorbate 20
`
`1
`0
`
`10
`0
`
`10
`0
`
`10
`1
`
`0
`
`1
`
`Storage for 3 Months at 25° C.
`
`Number of vials with
`particle formation after
`
`35
`
`40
`
`45
`
`Test sample
`
`7 days
`
`14 days 21 days 28 days
`
`Insulin glargine
`Insulin glargine + 0.010 mg/ml
`of polysorbate 20
`50 Insulin glargine + 0.015 mg/ml
`of polysorbate 20
`Insulin glargine + 0.020 mg/ml
`of polysorbate 20
`Insulin glargine + 0.030 mg/ml
`of polysorbate 20
`
`9
`2
`
`10
`2
`
`10
`2
`
`10
`2
`
`0
`
`0
`
`0
`
`0
`
`0
`
`Storage for 6 Months at 25° C.
`
`55
`
`60
`
`Number of vials with
`particle formation after
`
`Test sample
`
`7 days
`
`14 days 21 days 28 days
`
`Insulin glargine
`65 Insulin glargine + 0.010 mg/ml
`of polysorbate 20
`
`10
`0
`
`10
`0
`
`10
`0
`
`10
`1
`
`Mylan Ex.1092
`Mylan v. Sanofi - IPR2018-01676
`
`

`

`7
`
`-continued
`
`US 7,476,652 B2
`
`8
`Storage for 6 Months at 37° C.
`
`Number of vials with
`particle formation after
`
`5
`
`Number of vials with
`particle formation after
`
`Test sample
`
`7 days
`
`14 days 21 days 28 days
`
`Insulin glargine
`Insulin glargine + 0.010 mg/ml
`10 of polysorbate 20
`Insulin glargine + 0.015 mg/ml
`of polysorbate 20
`Insulin glargine + 0.020 mg/ml
`of polysorbate 20
`Insulin glargine + 0.030 mg/ml
`15 of polysorbate 20
`
`10
`0
`
`0
`
`0
`
`1
`
`10
`0
`
`0
`
`0
`
`1
`
`10
`0
`
`1
`
`0
`
`1
`
`10
`0
`
`0
`
`0
`
`1
`
`Test sample