U.S. patent application number 12/773356 was filed with the patent office on 2010-08-26 for acidic insulin preparations having improved stability.
This patent application is currently assigned to SANOFI-AVENTIS DEUTSCHLAND GMBH. Invention is credited to Anette BRUNNER-SCHWARZ, Norbert LILL.
Application Number | 20100216692 12/773356 |
Document ID | / |
Family ID | 29723248 |
Filed Date | 2010-08-26 |
United States Patent
Application |
20100216692 |
Kind Code |
A1 |
BRUNNER-SCHWARZ; Anette ; et
al. |
August 26, 2010 |
Acidic Insulin Preparations Having Improved Stability
Abstract
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 and
combinations thereof; a surfactant or combinations 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 combinations thereof, the pharmaceutical
formulation having a pH in the acidic range.
Inventors: |
BRUNNER-SCHWARZ; Anette;
(Frankfurt, DE) ; LILL; Norbert; (Kronberg,
DE) |
Correspondence
Address: |
ANDREA Q. RYAN;SANOFI-AVENTIS U.S. LLC
1041 ROUTE 202-206, MAIL CODE: D303A
BRIDGEWATER
NJ
08807
US
|
Assignee: |
SANOFI-AVENTIS DEUTSCHLAND
GMBH
Frankfurt am Main
DE
|
Family ID: |
29723248 |
Appl. No.: |
12/773356 |
Filed: |
May 4, 2010 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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12328208 |
Dec 4, 2008 |
7713930 |
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12773356 |
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11089777 |
Mar 25, 2005 |
7476652 |
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12328208 |
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10461740 |
Jun 13, 2003 |
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11089777 |
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60409338 |
Sep 9, 2002 |
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Current U.S.
Class: |
514/1.1 |
Current CPC
Class: |
A61P 3/06 20180101; A61K
9/0019 20130101; A61P 3/10 20180101; A61P 9/10 20180101; A61K 38/28
20130101; A61K 47/26 20130101; A61K 31/7032 20130101; A61P 5/50
20180101; A61K 31/7032 20130101; A61K 2300/00 20130101 |
Class at
Publication: |
514/3 |
International
Class: |
A61K 38/28 20060101
A61K038/28 |
Foreign Application Data
Date |
Code |
Application Number |
Jun 18, 2002 |
DE |
10227232.8 |
Claims
1. A pharmaceutical formulation comprising: (a) Gly(A21), Arg(B31),
Arg(B32)-human insulin; (b) polysorbate 20; (c) sodium chloride;
(d) glycerol; (e) m-cresol; and (f) water, wherein the
pharmaceutical formulation has a pH in the acidic range from 3.5 to
6.8.
2. The pharmaceutical formulation as claimed in claim 1, wherein
the Gly(A21), Arg(B31), Arg(B32)-human insulin is present at a
concentration of about 1.4 to about 35 mg per milliliter.
3. The pharmaceutical formulation as claimed in claim 1, wherein
the Gly(A21), Arg(B31), Arg(B32)-human insulin is present at a
concentration of about 3.6 mg per milliliter.
4. The pharmaceutical formulation as claimed in claim 3, further
including zinc.
5. The pharmaceutical formulation as claimed in claim 1, wherein
polysorbate 20 is present at a concentration of about 5 to 120
.mu.g per milliliter.
6. The pharmaceutical formulation as claimed in claim 1, wherein
polysorbate 20 is present at a concentration of about 20 to 75
.mu.g per milliliter.
7. The pharmaceutical formulation as claimed in claim 1, wherein
sodium chloride is present at a concentration of up to 150 mM.
8. The pharmaceutical formulation as claimed in claim 1, wherein
glycerol is present at a concentration of about 100 to 250 mM.
9. The pharmaceutical formulation as claimed in claim 1, further
comprising a buffer.
10. The pharmaceutical formulation as claimed in claim 9, wherein
the buffer is chosen from TRIS, phosphate, citrate, acetate, and
glycylglycine.
11. The pharmaceutical formulation as claimed in claim 10, wherein
said buffer is present in a concentration of 5-250 mM.
12. The pharmaceutical formulation as claimed in claimed 1, wherein
the pharmaceutical formulation has a pH in the acidic range from
3.5 to 4.5.
13. The pharmaceutical formulation as claimed in claimed 1, wherein
the pharmaceutical formulation has a pH of 4 (+/-0.2).
Description
[0001] This application is a continuation of U.S. application Ser.
No. 12/328,208, filed Dec. 4, 2008, now U.S. Pat. No. 7,713,930,
issued, May 11, 2010, which is a continuation of U.S. application
Ser. No. 11/089,777, filed Mar. 25, 2005, now U.S. Pat. No.
7,476,652, issued, Jan. 13, 2009, which is a continuation of Ser.
No. 10/461,740, filed Jun. 13, 2003, now abandoned; all of which
are incorporated herein by reference in their entirety which claims
the benefit of U.S. Provisional Application No. 60/409,338, filed
Sep. 9, 2002, and Federal Republic of Germany Application
10227232.8, filed Jun. 18, 2002.
SUMMARY OF THE INVENTION
[0002] 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 combinations 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 combinations 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 preparations in which a high stability to
thermal and/or physicomechanical 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 preparations.
BACKGROUND OF THE INVENTION
[0003] 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 insulin 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 combination with an oral
antidiabetic, is regarded as the most favorable form of
therapy.
[0004] 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 replacement 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 Complications Trial Research Group (1993) N. Engl. J.
Med. 329, 977-986) demonstrated clearly that chronically elevated
blood glucose levels are essentially responsible for the
development of diabetic late damage. Diabetic late damage is
microvascular and macrovascular damage which is manifested, under
certain circumstances, as retinopathy, nephropathy 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 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 formulations 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.
[0005] Insulin is a polypeptide of 51 amino acids, which are
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 used, but recently also insulin
derivatives and analogs.
[0006] 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 least one
amino acid residue from the corresponding, otherwise identical,
naturally occurring insulin. The amino acids can in this case also
be those which do not occur naturally.
[0007] Insulin derivatives are derivatives of naturally occurring
insulin or an insulin analog which are obtained by chemical
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 derivatives and insulin
analogs have a somewhat modified action compared with human
insulin.
[0008] 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 position B29 have various
amino acids, preferably proline, but not glutamic acid. EP 0 375
437 includes insulin analogs with lysine or arginine in B28, which
can optionally be additionally modified in B3 and/or A21.
[0009] In EP 0 419 504, insulin analogs are disclosed which are
protected against chemical modifications, in which asparagine in B3
and at least one further amino acid in the positions A5, A15, A18
or A21 are modified.
[0010] 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 this type have a
prolonged action. The insulin analogs described in EP-A 0 368 187
also have a delayed action.
[0011] 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
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 physiological as
possible can be established. Recombinant DNA 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 duration of action. Insulin glargine is injected as an
acidic, clear solution and precipitates on account of its solution
properties 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 (Schubert-Zsilavecz et
al., 2:125-130 (2001)).
[0012] The specific preparation of insulin glargine, which leads to
the prolonged duration of action, is characterized, in contrast to
previously described preparations, by a clear solution having 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)).
[0013] The proneness to aggregation can additionally be promoted 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.
[0014] WO 01/43762 describes aqueous, parenteral pharmaceutical
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.
[0015] WO 00/23098 describes insulin preparations stabilized using
polysorbate 20 or poloxamer 188 for pulmonary administration, but
does not describe the stabilization in an acidic solution against
aggregation nuclei.
[0016] 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.
[0017] The present invention was thus based on the object of
finding preparations for acid-soluble insulins containing
surfactants, 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
[0018] 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 under temperature stress.
[0019] 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.
[0020] The surfactants which can be used are, inter alia, nonionic
surfactants. In particular, pharmaceutically customary surfactants
are preferred, such as, for example: partial and fatty acid esters
and ethers of polyhydric alcohols such as of glycerol, sorbitol and
the like (SPAN.RTM., TWEEN.RTM., in particular TWEEN.RTM. 20 and
TWEEN.RTM. 80, MYRJ.RTM., BRIJ.RTM.), CREMOPHOR.RTM. or poloxamers.
The surfactants are present in the pharmaceutical composition in a
concentration of 5-200 .mu.g/ml, preferably of 5-120 .mu.g/ml and
particularly preferably of 20-75 .mu.g/ml.
[0021] The preparation can additionally optionally contain
preservatives (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.
[0022] Glycerol, dextrose, lactose, sorbitol and mannitol are
customarily present in the pharmaceutical preparation in a
concentration of 100-250 mM, NaCl in a concentration of up to 150
mM. Buffer substances, such as, for example, phosphate, acetate,
citrate, arginine, glycylglycine or TRIS (i.e.
2-amino-2-hydroxymethyl-1,3-propanediol) buffer and corresponding
salts, are present in a concentration of 5-250 mM, preferably
10-100 mM. Further excipients can be, inter alia, salts or
arginine.
[0023] The invention therefore relates to a pharmaceutical
formulation comprising a polypeptide selected from the group
consisting of insulin, an insulin analog, an insulin derivative, an
active insulin metabolite and combinations thereof; a surfactant 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 pharmaceutical formulation being a clear
solution which has a pH in the acidic range (pH 1-6.8), preferably
pH 3.5-6.8, very particularly preferably 3.5-4.5.
[0024] Preferred pharmaceutical formulations of the present
invention are those wherein the surfactant is selected from the
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.RTM.,
TWEEN.RTM., MYRJ.RTM., BRIJ.RTM., CREMOPHOR.RTM.; the polyols being
selected from the group consisting of polypropylene glycols,
polyethylene glycols, poloxamers, PLURONICS.RTM., and
TETRONICS.RTM.; the preservative being selected from the group
consisting of phenol, cresol, and parabens; the isotonicizing agent
being selected from the group consisting of mannitol, 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.sup.B28Pro.sup.B29 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)-human insulin; the insulin derivative being selected from
the group consisting of B29-N-myristoyl-des(B30) human insulin,
B29-N-palmitoyl-des(B30) human insulin, B29-N-myristoyl human
insulin, B29-N-palmitoyl human insulin, B28-N-myristoyl
Lys.sup.B28Pro.sup.B29 human insulin,
B28-N-palmitoyl-Lys.sup.B28Pro.sup.B29 human insulin,
B30-N-myristoyl-Thr.sup.B29Lys.sup.B30 human insulin,
B30-N-palmitoyl-Thr.sup.B29Lys.sup.B30 human insulin,
B29-N-(N-palmitoyl-.gamma.-glutamyl)-des(B30) human insulin,
B29-N-(N-lithocholyl-.gamma.-glutamyl)-des(B30) human insulin,
B29-N-(.omega.-carboxyheptadecanoyl)-des(B30) human insulin and
B29-N-(.omega.-carboxyheptadecanoyl) human insulin.
[0025] A further subject of the invention is a pharmaceutical
formulation such as described above, in which the insulin, the
insulin analog, the active insulin metabolite and/or the insulin
derivative is present in a concentration of 60-6000 nmol/ml,
preferably in a concentration of 240-3000 nmol/ml (this corresponds
approximately to a concentration of 1.4-35 mg/ml or 40-500
units/ml);
in which the surfactant is present in a concentration of 5-200
.mu.g/ml, preferably of 5-120 .mu.g/ml and particularly preferably
of 20-75 .mu.g/ml.
[0026] A further subject of the invention is a pharmaceutical
formulation such as mentioned above, in which glycerol and/or
mannitol is present in a concentration of 100-250 mM, and/or NaCl
is preferably present in a concentration of up to 150 mM.
[0027] A further subject of the invention is a pharmaceutical
formulation such as mentioned above, in which a buffer substance is
present in a concentration of 5-250 mM.
[0028] A further subject of the invention is a pharmaceutical
insulin 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.
[0029] 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.
[0030] A further subject of the invention is the use or the
addition of surfactants as stabilizer during the process for the
production of insulin, insulin analogs or insulin derivatives or
their preparations.
EXAMPLES
[0031] The following examples illustrate, but by no means limit,
the present invention.
[0032] Comparison investigations: Different preparations containing
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 injection, 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 concentration 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
specific amount of a surfactant is additionally added. The
solutions 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: [0033] 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.degree. 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 insulin syringe and discarded. At the
beginning and end of the working week this procedure is carried out
twice in order 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. [0034] 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.degree. 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
nephelometric units (formaldazine nephelometric unit=FNU). The
turbidity 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.RTM. 20)
[0035] a) The solution is sterile-filtered through a combination of
0.2 .mu.m and 0.1 .mu.m filters. It is then poured into 10 ml
injection vials and sealed using crimp caps having an inserted
sealing disk. 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.degree. C., 25.degree. C. and 37.degree. C. for a fixed period
of time.
[0036] 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.degree. C.
TABLE-US-00001 [0037] Number of vials with particle formation after
Test sample 7 days 14 days 21 days 28 days Insulin glargine 7 10 10
10 Insulin glargine + 0 0 0 0 0.010 mg/ml of polysorbate 20 Insulin
glargine + 0 0 0 0 0.015 mg/ml of polysorbate 20 Insulin glargine +
0 0 0 1 0.020 mg/ml of polysorbate 20 Insulin glargine + 0 0 0 0
0.030 mg/ml of polysorbate 20
Storage for 6 Months at 5.degree. C.
TABLE-US-00002 [0038] Number of vials with particle formation after
Test sample 7 days 14 days 21 days 28 days Insulin glargine 1 10 10
10 Insulin glargine + 0 0 0 1 0.010 mg/ml of polysorbate 20 Insulin
glargine + 0 0 0 0 0.015 mg/ml of polysorbate 20 Insulin glargine +
0 0 0 1 0.020 mg/ml of polysorbate 20 Insulin glargine + 0 0 1 0
0.030 mg/ml of polysorbate 20
Storage for 3 Months at 25.degree. C.
TABLE-US-00003 [0039] Number of vials with particle formation after
Test sample 7 days 14 days 21 days 28 days Insulin glargine 9 10 10
10 Insulin glargine + 2 2 2 2 0.010 mg/ml of polysorbate 20 Insulin
glargine + 0 0 0 1 0.015 mg/ml of polysorbate 20 Insulin glargine +
0 0 0 0 0.020 mg/ml of polysorbate 20 Insulin glargine + 0 0 0 0
0.030 mg/ml of polysorbate 20
Storage for 6 Months at 25.degree. C.
TABLE-US-00004 [0040] Number of vials with particle formation after
Test sample 7 days 14 days 21 days 28 days Insulin glargine 10 10
10 10 Insulin glargine + 0 0 0 1 0.010 mg/ml of polysorbate 20
Insulin glargine + 0 0 1 0 0.015 mg/ml of polysorbate 20 Insulin
glargine + 0 0 0 0 0.020 mg/ml of polysorbate 20 Insulin glargine +
0 0 0 0 0.030 mg/ml of polysorbate 20
Storage for 1 Month at 37.degree. C.
TABLE-US-00005 [0041] Number of vials with particle formation after
Test sample 7 days 14 days 21 days 28 days Insulin glargine 0 10 10
10 Insulin glargine + 0 3 3 5 0.010 mg/ml of polysorbate 20 Insulin
glargine + 0 0 0 0 0.015 mg/ml of polysorbate 20 Insulin glargine +
0 0 0 0 0.020 mg/ml of polysorbate 20 Insulin glargine + 0 0 0 0
0.030 mg/ml of polysorbate 20
Storage for 3 Months at 37.degree. C.
TABLE-US-00006 [0042] Number of vials with particle formation after
Test sample 7 days 14 days 21 days 28 days Insulin glargine 5 9 10
10 Insulin glargine + 1 1 1 1 0.010 mg/ml of polysorbate 20 Insulin
glargine + 0 0 0 0 0.015 mg/ml of polysorbate 20 Insulin glargine +
0 0 0 0 0.020 mg/ml of polysorbate 20 Insulin glargine + 0 0 0 0
0.030 mg/ml of polysorbate 20
Storage for 6 Months at 37.degree. C.
TABLE-US-00007 [0043] Number of vials with particle formation after
Test sample 7 days 14 days 21 days 28 days Insulin glargine 10 10
10 10 Insulin glargine + 0 0 0 0 0.010 mg/ml of polysorbate 20
Insulin glargine + 0 0 1 0 0.015 mg/ml of polysorbate 20 Insulin
glargine + 0 0 0 0 0.020 mg/ml of polysorbate 20 Insulin glargine +
1 1 1 1 0.030 mg/ml of polysorbate 20
[0044] Without addition of polysorbate 20, particle formation can
occur in the solution even after 7 days in use. By addition of
polysorbate 20, the particle formation can be markedly suppressed
during the in use period.
[0045] The stabilizing action of polysorbate 20 is retained even on
storage at elevated temperatures for a period of 3 months.
[0046] A decline in the stabilizing action due to possible
hydrolysis of the polysorbate in the acidic medium of the solution
cannot be determined in comparison with the data after storage for
1 month.
Example 2
Stabilization of Insulin Glargine Using Polysorbate 20 Under
Physico-Mechanical Stress Loading
[0047] a) The solution is sterile-filtered through a combination of
0.2 .mu.m und 0.1 .mu.m filters. It is then poured into 10 ml
injection vials and sealed using crimp caps having an inserted
sealing disk. b) A comparison solution is prepared identically, but
first a suitable amount of surfactant (0.010-0.030 mg/ml of
polysorbate 20) is suspended in water for injection.
[0048] The samples are stored at +5.degree. C., 25.degree. C. und
37.degree. C. for a fixed period of time. 5 samples in each case
are then subjected to a shaking test. The results are shown in the
table below, the limit 15 FNU corresponds to turbidities which are
discernible in daylight.
Storage for 1 Month at 5.degree. C.
TABLE-US-00008 [0049] Number of vials >15 FNU 0 0.5 1 2 3 4 6
Test sample days days day days days days days 8 days 10 days
Insulin glargine 0 0 0 2 3 3 4 4 4 Insulin glargine + 0 0 0 0 0 1 3
4 5 0.010 mg/ml of polysorbate 20 Insulin glargine + 0 0 0 0 0 0 0
0 0 0.015 mg/ml of polysorbate 20 Insulin glargine + 0 0 0 0 0 0 0
0 0 0.020 mg/ml of polysorbate 20 Insulin glargine + 0 0 0 0 0 0 0
0 0 0.030 mg/ml of polysorbate 20
Storage for 1 Month at 25.degree. C.
TABLE-US-00009 [0050] Number of vials >15 FNU 0 0.5 1 2 3 4 6
Test sample days days day days days days days 8 days 10 days
Insulin glargine 0 0 0 1 1 1 1 2 3 Insulin glargine + 0 0 0 0 0 0 1
2 3 0.010 mg/ml of polysorbate 20 Insulin glargine + 0 0 0 0 0 0 0
0 0 0.015 mg/ml of polysorbate 20 Insulin glargine + 0 0 0 0 0 0 0
0 0 0.020 mg/ml of polysorbate 20 Insulin glargine + 0 0 0 0 0 0 0
0 0 0.030 mg/ml of polysorbate 20
Storage for 1 Month at 37.degree. C.
TABLE-US-00010 [0051] Number of vials >15 FNU 0 0.5 1 2 3 4 6
Test sample days days day days days days days 8 days 10 days
Insulin glargine 0 0 0 2 5 5 5 5 5 Insulin glargine + 0 0 0 0 0 0 0
0 0 0.010 mg/ml of polysorbate 20 Insulin glargine + 0 0 0 0 0 0 0
0 0 0.015 mg/ml of polysorbate 20 Insulin glargine + 0 0 0 0 0 0 0
0 0 0.020 mg/ml of polysorbate 20 Insulin glargine + 0 0 0 0 0 0 0
0 0 0.030 mg/ml of polysorbate 20
[0052] Without addition of polysorbate 20, even after 2 days of
severe physicomechanical stress, a visible turbidity can occur in
the solution. By addition of polysorbate 20, the formation of
turbidity during physicomechanical stressing can be markedly
delayed. The stabilizing action of polysorbate 20 is retained even
on storage at elevated temperatures.
[0053] A decline in the stabilizing action due to possible
hydrolysis of the polysorbate in the acidic medium of the solution
cannot be detected.
Example 3
Comparison of the Stabilization of the in Use Period of Insulin
Glargine Using Polysorbate 20 (Tween.RTM. 20) and Using Polysorbate
80 (Tween.RTM. 20)
[0054] Open 10 vials in each case to give 5 ml of insulin glargine
injection solution and
a) addition of 0.001 mg/ml of polysorbate 20 b) addition of 0.01
mg/ml of polysorbate 20 c) addition of 0.001 mg/ml of polysorbate
80 d) addition of 0.01 mg/ml of polysorbate 80 in the form of a
concentrated stock solution.
[0055] The samples are then subjected to an in use test.
[0056] The results are shown in the table below.
TABLE-US-00011 Vials with particle formation after Test sample 7
days 14 days 21 days 28 days Insulin glargine + no yes Yes, Yes,
0.001 mg/ml of particles particles polysorbate 20 increasingly
increasingly occur occur Insulin glargine + no no no no 0.010 mg/ml
of polysorbate 20 Insulin glargine + no yes Yes, Yes, 0.001 mg/ml
of particles particles polysorbate 80 increasingly increasingly
occur occur Insulin glargine + no no no no 0.010 mg/ml of
polysorbate 80
[0057] An addition of polysorbate 20 or of polysorbate 80 in a
concentration of 0.001 mg/ml are equally able to stabilize the
solution against particle formation during the in use period.
* * * * *