U.S. patent application number 10/429508 was filed with the patent office on 2004-01-08 for novel formulations.
Invention is credited to Langkjaer, Liselotte.
Application Number | 20040006000 10/429508 |
Document ID | / |
Family ID | 29414624 |
Filed Date | 2004-01-08 |
United States Patent
Application |
20040006000 |
Kind Code |
A1 |
Langkjaer, Liselotte |
January 8, 2004 |
Novel formulations
Abstract
Stable insulin formulations can be prepared by mixing a
monomeric insulin and a soluble acylated insulin analog.
Inventors: |
Langkjaer, Liselotte;
(Holte, DK) |
Correspondence
Address: |
Reza Green, Esq.
Novo Nordisk Pharmaceuticals, Inc.
100 College Road West
Princeton
NJ
08540
US
|
Family ID: |
29414624 |
Appl. No.: |
10/429508 |
Filed: |
May 5, 2003 |
Current U.S.
Class: |
514/6.5 ;
514/5.9; 514/6.9 |
Current CPC
Class: |
A61K 38/28 20130101;
A61P 3/10 20180101; A61P 5/50 20180101 |
Class at
Publication: |
514/3 |
International
Class: |
A61K 038/28 |
Foreign Application Data
Date |
Code |
Application Number |
May 7, 2002 |
DK |
PA 2002 00683 |
Claims
1. A pharmaceutical soluble formulation comprising (i) a soluble
acylated insulin analogue and (ii) a monomeric insulin or human
insulin, wherein the lower limit of the molar ratio between the
soluble acylated insulin analogue and the monomeric insulin or
human insulin is 7:93, and the upper limit of the molar ratio
between the soluble acylated insulin analogue and the monomeric
insulin or human insulin is 57:43.
2. A formulation according to claim 1, wherein the lower limit is
11:89 and the upper limit is 41:59.
3. A pharmaceutical soluble formulation comprising (i) a soluble
acylated insulin analogue and (ii) a monomeric insulin or human
insulin, wherein the ratio between the soluble acylated insulin
analogue and the monomeric insulin or human insulin is in the range
from about 7:93 to about 57:43 on a mole to mole basis, with the
proviso that if the soluble acylated insulin analogue is insulin
detemir and the monomeric insulin is insulin aspart then the ratio
between detemir and aspart is not more than about 41:59 on a mole
to mole basis.
4. The formulation according to claim 1, wherein the pH of the
formulation is in the range from about 7 to about 8.
5. The formulation according to claim 1, which is suitable for use
in a continuous infusion system.
6. The formulation according to claim 1, further comprising one or
more of: an agent rendering the solution isotonic, an antimicrobial
preservative, a pH-buffering agent, and a suitable zinc salt.
7. The formulation according to claim 1, wherein the soluble
acylated insulin analogue is insulin detemir.
8. The formulation according to claim 1, comprising human
insulin.
9. The formulation according to claim 1, wherein the monomeric
insulin is insulin aspart.
10. The formulation according to claim 1, wherein the monomeric
insulin is insulin lispro.
11. The formulation according to claim 1, wherein the monomeric
insulin is insulin glulisine (Lys.sup.B3,Glu.sup.B29 human
insulin).
12. The formulation according to claim 1, wherein the monomeric
insulin is different from insulin aspart, insulin lispro, and
insulin glulisine (Lys.sup.B3,Glu.sup.B29 human insulin).
13. The formulation according to claim 1, wherein the concentration
of insulin is between about 10 U/ml and 1500 U/ml.
14. The formulation according to claim 1, wherein the preservative
is phenol, m-cresol or a mixture of phenol and m-cresol.
15. The formulation according to claim 1, wherein the concentration
of phenol and/or m-cresol is in the range from about 20 mM to about
50 mM.
16. The formulation according to claim 1, comprising zinc ions in
an amount corresponding to from about 2.3 to about 4.5 Zn.sup.2+
per insulin hexamer.
17. The formulation according to claim 1, wherein the zinc salt is
zinc chloride, zinc oxide, or zinc acetate.
18. The formulation according to claim 1, containing halogenide
ions at a concentration from about 1 to about 100 mM.
19. The formulation according to claim 1, containing as isotonic
agent glycerol, mannitol, sorbitol, or a mixture thereof in a
concentration in the range from about 100 to about 250 mM.
20. The formulation according to claim 1, wherein the pH-buffer is
sodium phosphate, TRIS (trometamol), N-glycylglycine, or
L-arginine.
21. The formulation according to claim 1, wherein the pH-buffer is
a physiologically acceptable buffer in a concentration in the range
from about 3 mM to about 20 mM.
22. The formulation according to claim 1, comprising: (i) insulin
aspart (ii) insulin detemir, wherein the concentrations of (i) and
(ii) are suitable for continuous infusion systems; and (iii) a
phenolic preservative at a concentration of about 30 mM to about 40
mM; (iv) glycerol in a concentration of about 0.17 M; (v) dibasic
sodium phosphate at concentration of about 5 mM to about 7 mM; (vi)
zinc ions in an amount corresponding to about 2.5 to about 3.5
Zn.sup.2+/hexamer insulin or from about 0.4 to about 0.6
Zn.sup.2+/monomer insulin.
23. The use of a soluble acylated insulin analogue in an amount in
the range with the lower limit of the molar ratio between the
soluble acylated insulin analogue and the monomeric insulin or
human insulin being 7%, preferably 11%, more preferred 14%, and the
upper limit of the molar ratio between the soluble acylated insulin
analogue and the monomeric insulin or human insulin being 57%,
preferably 41%, more preferred 31%, preferably 24% and even more
preferred 20%, of the total amount of insulin to improve the
physical stability in an aqueous solution containing a monomeric
insulin or human insulin.
24. The use, according to any one of the previous use claims,
wherein the improvement in physical stability is a reduction or
avoidance of fibrillation.
25. The use, according to the previous claim, wherein the stability
factor, when determined by the test described in example 1 above,
of the formulation is above about 2.5, preferably above about
4.
26. The use of a soluble acylated insulin analogue in an amount in
the range with the lower limit of the molar ratio between the
soluble acylated insulin analogue and the monomeric insulin or
human insulin being 7%, preferably 11%, more preferred 14%, and the
upper limit of the molar ratio between the soluble acylated insulin
analogue and the monomeric insulin or human insulin being 57%,
preferably 41%, more preferred 31%, preferably 24% and even more
preferred 20%, of the total amount of insulin to reduce the risk
for ketoacidosis in an aqueous solution containing a monomeric
insulin or human insulin.
27. The use, according to any one of the two preceding claims,
which is characterized by any of the features mentioned
specifically in any of the above sub claims to pharmaceutical
compositions.
28. A method of treating diabetes in a patient in need of such
treatment, comprising administering to the patient a
therapeutically effective amount of a pharmaceutical formulation
according to claim 1.
29. A method, according to the previous claims, which is
characterized by any of the features mentioned specifically in any
of the above sub claims to pharmaceutical compositions.
30. A reservoir in a continuous infusion system comprising a
soluble acylated insulin analog.
31. A reservoir in a continuous infusion system comprising a
pharmaceutical formulation according to claim 1.
32. The reservoir, according to claim 31, which is characterized by
any of the features mentioned specifically in any of the above sub
claims to pharmaceutical compositions.
33. Any novel feature or combination of features described
herein.
34. A method for stabilizing a pharmaceutical formulation, said
method comprising: (i) providing as a first component of said
formulation a monomeric insulin and/or human insulin, and (ii)
adding as a second component of said formulation a soluble acylated
insulin analogue, wherein the lower limit of the molar ratio
between the soluble acylated insulin analogue and the monomeric
insulin or human insulin is 7:93, and the upper limit of the molar
ratio between the soluble acylated insulin analogue and the
monomeric insulin or human insulin is 57:43.
Description
FIELD OF THE INVENTION
[0001] This invention relates to a pharmaceutical formulation with
improved physical stability being a mixture of a soluble acylated
insulin analogue and a monomeric insulin or human insulin,
preferably for use in an infusion system. Furthermore, this
invention relates to the additional aspects mentioned in the claims
below.
[0002] The object of this invention is to overcome or ameliorate at
lest some of the disadvantages of the prior art. Hence, the more
specific objects mentioned below are more or less fulfilled.
BACKGROUND OF THE INVENTION
[0003] Diabetes is a general term for disorders in man having
excessive urine excretion as in diabetes mellitus and diabetes
insipidus. Diabetes mellitus is a metabolic disorder in which the
ability to utilize glucose is partly or completely lost. About 5%
of all people suffer from diabetes.
[0004] Since the introduction of insulin in the 1920's, continuous
strides have been made to improve the treatment of diabetes
mellitus. To help avoid extreme glycemia levels, diabetic patients
often practice multiple daily injection therapy, whereby monomeric
insulin is administered with each meal and acylated or intermediate
acting insulin is administered once or twice daily to cover the
basal need.
[0005] In the treatment of diabetes mellitus, many varieties of
insulin formulations have been suggested and used, such as regular
insulin, isophane insulin (designated NPH), insulin zinc
suspensions (such as Semilente.RTM., Lente.RTM., and
Ultralente.RTM.), and biphasic isophane insulin. As diabetic
patients are treated with insulin for several decades, there is a
major need for safe and life quality improving insulin
formulations. Some of the commercial available insulin formulations
are characterized by a fast onset of action and other formulations
have a relatively slow onset but show a more or less prolonged
action. Fast-acting insulin formulations are usually solutions of
insulin, while retarded acting insulin formulations can be
suspensions containing insulin in crystalline and/or amorphous form
precipitated by addition of zinc salts alone or by addition of
protamine or by a combination of both. In addition, some patients
are using formulations having both a fast onset of action and a
more prolonged action. Such a formulation may be an insulin
solution wherein protamine insulin crystals are suspended. Some
patients do themselves prepare the final formulation by mixing an
insulin solution with an insulin suspension formulation in the
ratio desired by the patient in question.
[0006] Human insulin consists of two polypeptide chains, the
so-called A and B chains which contain 21 and 30 amino acid
residues, respectively. The A and B chains are interconnected by
two cystine disulphide bridges. Insulin from most other species has
a similar construction, but may not contain the same amino acid
residues at the same positions.
[0007] The development of the process known as genetic engineering
has made it possible to prepare a great variety of insulin
compounds being analogous to human insulin. In these insulin
analogues, one or more of the amino acids have been substituted
with other amino acids which can be coded for by the nucleotide
sequences.
[0008] Normally, insulin formulations are administered by
subcutaneous injection. What is important for the patient, is the
action profile of the insulin formulation which is the action of
insulin on the glucose metabolism as a function of the time from
the injection. In this profile, inter alia, the time for the onset,
the maximum value, and the total duration of action are important.
A variety of insulin formulations with different action profiles
are desired and requested by the patients. One patient may, on the
same day, use insulin formulations with very different action
profiles. The action profile requested is, for example, depending
on the time of the day and the amount and composition of any meal
eaten by the patient.
[0009] Stable insulin formulations are particularly required for
use in delivery devices that expose these agents to elevated
temperatures and/or mechanical stress. For example, stable insulin
formulations are required for use in continuous infusion systems
and pen delivery devices.
[0010] There is a need for new ways of stabilizing since a
stablizer, Genapol.RTM. (poloxamer 171), which for a long period of
time has been used for stabilizing human insulin for pumps, may
have some undesired effects (see Diabetes Metab. 26 (2000),
304-306).
[0011] In continuous infusion systems, a fluid containing an
insulin formulation is pumped from a reservoir, usually to a
subcutaneous, intravenous, or intraperitoneal depot. The reservoir,
which must be exchanged or refilled periodically, is attached to
the patient's body, or implanted into the patient's body. In either
case, the patient's body heat and body motion, plus turbulence in
the tubing and pump impart a relatively high amount of
thermo-mechanical energy to the formulation. In the interest of
minimizing the frequency with which the reservoir is filled, and of
minimizing the size of the reservoir, formulations having a
relatively high concentration of insulin are highly advantageous.
It is desirable to have insulin formulations which are stable for
at least one month under stressful in-use conditions.
[0012] Formulations of insulin for use in continuous infusion
systems must remain soluble and substantially free of aggregation,
even though subjected to the patient's body heat and motion for
periods ranging from a few days to several months. Instability is
promoted by the thermo-mechanical stress to which formulations are
exposed in continuous infusion systems. Therefore, improvements in
the physical stability of concentrated insulin formulations is
urgently needed to permit them to be used successfully in
continuous infusion systems.
[0013] It has become usual to use monomeric insulins in pumps.
However, compared with human insulin, monomeric insulins have an
increased tendency to form insoluble fibrils.
[0014] Among others, there are two major problems in connection
with the use of insulin formulations in a continuous infusion
system:
[0015] 1. Due to fibrillation of the insulin component, the
catheter may clog, and
[0016] 2. There is a risk of fast development of ketoacidosis which
may be fatal. Ketoacidosis may result from discontinuation of
insulin delivery, for example due to fibrillation, pump failure, or
the patient forgetting to reapply the pump after disconnection.
[0017] According to U.S. Pat. No. 4,476,118, pharmaceutical
solutions of dissolved insulin having improved physical stability
can be prepared by using ionized zinc salts. These solutions are
particularly adapted for use in continuous insulin delivery
equipment.
[0018] According to U.S. Pat. No. 4,472,385, pharmaceutical
solutions of dissolved insulin having improved physical stability
particularly adapted for use in continuous insulin delivery
equipment can be by using a calcium or magnesium salt.
[0019] According to U.S. Pat. No. 4,614,730, insulin solutions may
be stabilized by using a phospholipid.
[0020] According to U.S. Pat. No. 5,866,538, insulin formulations
of superior chemical stability can be obtained in the presence of
glycerol and/or mannitol and rather low halogenide
concentrations.
[0021] One object of this invention is to furnish insulin
formulations especially suited for use in an infusion system.
[0022] Another object of this invention is to furnish insulin
formulations having superior long-term physical stability when
exposed to high mechanical energy input.
[0023] Another object of this invention is to furnish insulin
formulations having superior long-term physical stability when
exposed to high temperature.
[0024] Another object of this invention is to furnish insulin
formulations having superior long-term physical stability when
exposed to high temperature and mechanical energy input.
[0025] Another object of this invention is to furnish insulin
formulations having a low tendency of fibrillation.
[0026] Another object of this invention is to furnish insulin
formulations having a convenient profile of action.
[0027] Another object of this invention is to furnish insulin
formulations having both a fast on-set of action and also a
retarded action.
[0028] Another object of this invention is to furnish insulin
formulations having no or only a minor amount of non-dissolved
material.
SUMMARY OF THE INVENTION
DETAILED DESCRIPTION OF THE INVENTION
[0029] Definitions
[0030] The term "amino acid" as used herein, refers to amino acids
which can be coded for by nucleotide sequences. Analogously, this
applies to the term amino acid residue which is an amino acid from
which hydroxy has been removed from a carboxyl group and/or
hydrogen has been removed from an amino group. Similarly, a peptide
and a peptide residue consists of amino acid residues.
[0031] The term "human insulin analogue" as used herein, refers to
human insulin wherein one or more of the amino acid residues have
been exchanged with another amino acid residue and/or from which
one or more amino acid residue has been deleted and/or from which
one or more amino acid residue has been added. These human insulin
analogues have an anti-diabetic activity sufficiently high to be
used to treat diabetic patients.
[0032] The term "acylated insulin analogue", when used herein,
refers to human insulin or a human insulin analogue having one or
more lipophilic substituents. The preferred lipophilic substituents
are acyl groups. Examples of acylated insulin analogues are
described in WO 95/07931 and WO 96/29344, more precisely in claim 1
thereof. These publications are hereby incorporated by reference.
An example of a specific acylated insulin analogue is insulin
detemir (i.e., LYS.sup.B29(N.sup..epsilon.-te- tradecanoyl)
des(B30) human insulin).
[0033] The term "monomeric insulin", when used herein, refers to
human insulin analogs that are less prone to self-association (into
dimers and hexamers) than human insulin. For more information,
reference is made to Diabetes Care 13 (1990), 923-954, which is
incorporated by reference. Examples of monomeric insulins are human
insulin wherein Pro at position B28 is replaced by Asp, Lys, Leu,
Val, or Ala; Lys at position B29 optionally is replaced by Pro; the
amino acids at positions B28-B30 are deleted; or the amino acid at
position B27 is deleted. A more detailed description of such
monomeric compounds can be found in U.S. patent application Ser.
No. 07/388,201 and European patent publication number 383,472 and
214,826 A2, all of which are hereby incorporated by reference. One
skilled in the art would recognize that other modifications to the
monomeric insulins are possible. Such modifications are widely
accepted in the art and, compared with human insulin, they include
replacement of Phe at position B1 with Asp; replacement of Thr at
position B30 with Ala; replacement of Ser at position B9 with Asp;
deletion of the amino acid at position B1; optionally, deletion of
Thr at position B2; or deletion of the amino acid at position B30.
Particularly preferred monomeric insulins are insulin lispro
(Lys.sup.B28,Pro.sup.B29 human insulin) and insulin aspart
(Asp.sup.B28 human insulin). A further monomeric insulin is insulin
glulisine (Lys.sup.B3, Glu.sup.B29 human insulin), vide
Diabetologia 42, suppl. 1, page A178, abstract # 665.
[0034] Insulin means human insulin, human insulin analogs,
monomeric insulin plus acylated insulin analogues.
[0035] The term "infusion system", when used herein, refers to a
device for continuously administering a fluid to a patient
parenterally for an extended period of time or for intermittently
administering a fluid to a patient parenterally over an extended
period of time without having to establish a new site of
administration each time the fluid is administered. The fluid
contains a compound having insulin activity. The device comprises a
reservoir for storing the fluid before it is infused, a pump, a
catheter, or other tubing for connecting the reservoir to the
administration site via the pump, and control elements to regulate
the pump. The device may be constructed for implantation, usually
into the peritoneum. In such a case, the insulin reservoir will
usually be adapted for percutaneous refilling. Obviously, when the
device is implanted, the contents of the reservoir will be at body
temperature, and subject to the patient's body motion.
[0036] The term "fibrillation", when used herein, refers to a
physical process by which partially unfolded insulin molecules
interact with each other to form insoluble linear aggregates. Under
the influence of heat and exposure to hydrophobic surfaces, insulin
undergoes conformational changes, resulting in successive, linear
aggregation and formation of a viscous gel or insoluble
precipitates. The degree of fibrillation can be determined as
described in the stress test, below. A more detailed explanation is
given in J. Pharm. Science. 86 (1997), 517-525, which is
incorporated by reference.
[0037] The term "U", when used herein, refers to insulin units.
Most of the currently used (marketed) insulins (bovine, porcine,
human, lispro, aspart, and glargine) have a potency of one unit
which equals 6 nmol. Long-acting acylated insulins have reduced
potency compared to human insulin. Thus, for insulin detemir one
unit corresponds to 24 nmol. For other insulins, the relation
between U and nmol can be determined, if not known already, for
example, by determining the amount giving a similar pharmacological
(blood glucose lowering) effect as that of human insulin.
[0038] The content of zinc is expressed per hexamer insulin as a
theoretical value, i.e., as the number of zinc atoms per 6
molecules of monomer insulin, independent on whether all hexamer
insulin actually is present as hexamer insulin or not.
[0039] Stabilization of a formulation as used herein refers to a
reduction in the formation of fibrils or other aggregates or
maintenance of overall solubility and/or chemical integrity and
corresponding maintenance of pharmacological efficacy. For example,
a stable formulation according to the invention is one that can be
stored at 4.degree. C. without forming fibrils after at least about
10 days, preferably at least about 20 days, more preferably at
least about 50 days, and, most preferably, at least about 100 days;
or that can be stored at 25.degree. C. without forming fibrils
after at least about 2 days, preferably at least about 5 days, more
preferably at least about 10 days, most preferably at least about
25 days.
DESCRIPTION OF THE INVENTION
[0040] This invention relates to mixtures of a soluble acylated
insulin analog and a monomeric insulin or human insulin. It has,
surprisingly, been found that such formulations have a
substantially improved stability, for example, when exposed to
physical stress. For example, the formulations of this invention
have a low tendency to fibrillation. The formulations of this
invention contain no or only a minor amount of non-dissolved
material. Furthermore, said formulations give both a fast onset of
action and also a retarded action. In addition to this, said
formulations have convenient profiles of action.
[0041] The use of the formulation of this invention reduces the
risk of ketoacidosis since, contrary to the usual insulin
treatments via a pump, the patient gets an amount of a long-acting
insulin, i.e., an acylated insulin.
[0042] In a broad aspect, this invention relates to a
pharmaceutical soluble formulation comprising a soluble acylated
insulin analogue and a monomeric insulin or human insulin wherein
the lower limit of the molar ratio between the soluble acylated
insulin analogue and the monomeric insulin or human insulin is
7:93, preferably 11:89, more preferred 14:86, and the upper limit
of the molar ratio between the soluble acylated insulin analogue
and the monomeric insulin or human insulin is 57:43, preferably
41:59, more preferred 31:69, preferably 24:76 and even more
preferred 20:80.
[0043] In a preferred embodiment of this invention, the
pharmaceutical soluble formulation according to this invention
comprises a soluble acylated insulin analogue and a monomeric
insulin or human insulin wherein the ratio between the soluble
acylated insulin analogue and the monomeric insulin or human
insulin is in the range from about 7:93 to about 57:43 on a mole to
mole basis (corresponding to a content of 7-57% on a mole to mole
basis). Since the skilled art worker, especially physicians, are
more familiar with U (units) than moles, it can be added that if
the soluble acylated insulin analogue and the monomeric insulin or
human insulin have the same strength, the content of the soluble
acylated insulin analogue will be 7-57% on a unit to unit basis.
If, however, the strength of the soluble acylated insulin analogue
is only, say, 25% of the strength of the monomeric insulin or human
insulin, then the pharmaceutical soluble formulation according to
this invention comprises a soluble acylated insulin analogue and
human insulin or a monomeric insulin wherein the ratio between the
soluble acylated insulin analogue and the monomeric insulin or
human insulin is in the range from about 2:98 to about 25:75, on a
unit to unit basis, preferably in the range from about 3:97 to
about 15:85, on a unit to unit basis, more preferred in the range
from about 4:96 to about 10:90, on a unit to unit basis.
[0044] According to one embodiment, the present invention does not
cover a pharmaceutical soluble formulation comprising insulin
aspart and insulin detemir wherein the ratio between insulin aspart
and insulin detemir is in the range from 15:85 to 85:15, on a unit
to unit basis, corresponding to the ratio between detemir and
aspart being not more than about 41:59 on a mole to mole basis,
vide our Danish patent application No. PA 2002 00684 the content of
which is hereby incorporated by reference.
[0045] According to another embodiment, the present invention
relates to a pharmaceutical soluble formulation comprising a
soluble acylated insulin analogue and a monomeric insulin or human
insulin wherein the molar ratio between the soluble acylated
insulin analogue and the monomeric insulin or human insulin is in
the range from about 11:89 to about 41:59, preferably in the range
from about 14:86 to about 31:69.
[0046] In some embodiments, the formulations of the present
invention exhibit a stability that is enhanced relatively to
formulations of monomeric insulin or human insulin alone, i.e,
without the soluble acylated insulin component. For example, the
stability may be enhanced 2-fold, preferably 3-fold, more
preferably 5-fold, even more preferably 10-fold or most preferably
more than 10-fold.
[0047] In assessing the stability under stressful conditions,
fibril formation can be measured visually, by conventional
spectroscopic means, or by, e.g.thioflavin T fluorescence
spectroscopy (Nielsen et al.: Biochemistry 40 (2001) p.
8397-8409).
[0048] For example, a formulation according to the invention that
exhibits a 2-fold enhanced stability relative to a reference
formulation is a formulation that, e.g., must be stored twice as
long as the reference formulation before fibril formation can be
detected.
[0049] In a preferred embodiment of this invention, the formulation
is suitable for use in a continuous infusion pump. Hence, in one
aspect, the present invention relates to a reservoir in a
continuous infusion system comprising a soluble acylated insulin
analogue as mentioned herein. In another aspect, this invention
relates to a reservoir containing a pharmaceutical soluble
formulation comprising a soluble acylated insulin analogue and a
monomeric insulin or human insulin wherein the lower limit of the
molar ratio between the soluble acylated insulin analogue and the
monomeric insulin or human insulin is 7:93, preferably 11:89, more
preferred 14:86, and the upper limit of the molar ratio between the
soluble acylated insulin analogue and the monomeric insulin or
human insulin is 57:43, preferably 41:59, more preferred 31:69,
preferably 24:76 and even more preferred 20:80.
[0050] The pharmaceutical formulation of this invention may be
prepared using the conventional techniques of the pharmaceutical
industry which involves dissolving and mixing the pertinent
ingredients as appropriate to give the desired end product.
[0051] Hence, using a soluble acylated insulin analogue in an
amount from about 1% to about 15% of the total activity of the
insulin calculated in insulin units, it is possible to prepare a
medicament having improved physical stability in an aqueous
solution containing a monomeric insulin or human insulin.
[0052] Thus, according to one procedure, on one hand, a soluble
acylated insulin analogue and, on the other hand, a monomeric
insulin or human insulin is dissolved in an amount of water, the
total volume of which is somewhat less than the final volume of the
formulation to be prepared. An isotonic agent, a preservative and a
buffer is added as required and the pH value of the solution is
adjusted--if necessary--using an acid, for example, hydrochloric
acid, or a base, for example, aqueous sodium hydroxide as needed.
Finally, the volume of the solution is adjusted with water to give
the desired concentration of the ingredients.
[0053] In a preferred embodiment of this invention, the soluble
acylated insulin analogue is insulin detemir
(Lys.sup.B29(N.sup..epsilon.-tetradec- anoyl) des( B30) human
insulin). In a further preferred embodiment of this invention, the
acylated insulin is Lys.sup.B29(N.sup..epsilon.-hexadecano- yl)
des(B30) human insulin; Lys.sup.B29(N.sup..epsilon.-tetradecanoyl)
human insulin; Lys.sup.B29(N.sup..epsilon.-hexadecanoyl) human
insulin; Lys.sup.B28 (N.sup..epsilon.-tetradecanoyl) Lys.sup.B28
Pro.sup.B29 human insulin;
Lys.sup.B28(N.sup..epsilon.-hexadecanoyl) Lys.sup.B28Pro.sup.B29
human insulin; Lys.sup.B30(N.sup..epsilon.-tetradecanoyl)
Thr.sup.B29Lys.sup.B30 human insulin;
Lys.sup.B30(N.sup..epsilon.-hexadec- anoyl) Thr.sup.B29Lys.sup.B30
human insulin; Lys.sup.B29(N.sup..epsilon.-(-
N-hexadecanoyl-.gamma.-glutamyl)) des(B30) human insulin;
Lys.sup.B29(N.sup..epsilon.-(N-lithocholyl-.gamma.-glutamyl))
des(B30) human insulin;
Lys.sup.B29(N.sup..epsilon.-(.omega.-carboxyheptadecanoyl)- )
des(B30) human insulin; or
Lys.sup.B29(N.sup..epsilon.-(.omega.-carboxyh- eptadecanoyl)) human
insulin.
[0054] In a preferred embodiment of this invention, human insulin
is used. In another preferred embodiment of this invention, the
monomeric insulin is insulin aspart. In a further preferred
embodiment, the monomeric insulin is insulin lispro. In a still
further preferred embodiment, the monomeric insulin is Lys.sup.B3,
Glu.sup.B29 human insulin.
[0055] In a further preferred embodiment of this invention, the
release of insulin activity from the formulation of this invention,
after parenteral administration thereof to a human being within the
first 4 hours, is at least about 50%.
[0056] In a preferred embodiment of this invention, the formulation
contains an agent rendering the solution isotonic, an antimicrobial
preservative, a pH-buffering agent, and a suitable zinc salt. In a
preferred embodiment, the formulation has a pH value in the range
from about 7 to about 8.
[0057] In a preferred embodiment of this invention, the formulation
has a total amount of the insulin in the range with the lower limit
being 10 U/ml, preferably 40 U/ml, more preferred 100 U/ml, and
even more preferred 150 U/ml, and the upper limit being 1500 U/ml,
preferably 1000 U/ml, more preferred 500 U/ml.
[0058] In another preferred embodiment of this invention, the
formulation has a total amount of the insulin in the range from
about 10 U/ml to about 1500 U/ml, preferably in the range from
about 40 U/ml to about 1000 U/ml, more preferred in the range from
about 100 U/ml to about 500 U/ml, for example, 100, 200, 400, or
500 U/ml.
[0059] In a preferred embodiment of this invention, the
preservative is phenol, m-cresol or a mixture of phenol and
m-cresol. In a further preferred embodiment of this invention, the
total concentration of phenol and/or m-cresol is in the range from
about 20 mM to about 50 mM, preferably in the range from about 30
mM to about 45 mM. The concentration of phenol and/or m-cresol is,
inter alia, depending on the concentration of insulin.
[0060] In a preferred embodiment of this invention, the formulation
has a content of zinc ions at the disposal of insulin in
proportions in the range from about 2.3 to about 4.5 Zn.sup.2+ per
hexamer insulin (corresponding to from about 0.38 to about 0.75
Zn.sup.2+/monomer insulin). The zinc salt used for preparing the
formulations of this invention may, for example, be zinc chloride,
zinc oxide or zinc acetate.
[0061] In a preferred embodiment of this invention, the isotonic
agent is glycerol, mannitol, sorbitol or a mixture thereof at a
concentration in the range from about 100 to 250 mM.
[0062] In another preferred embodiment of this invention, the
formulation contains halogenide ions, preferably as sodium
chloride, in an amount corresponding to from about 1 mM to about
100 mM, preferably from about 5 mM to about 40 mM.
[0063] In a preferred embodiment of this invention, the pH buffer
is sodium phosphate, TRIS (trometamol), N-glycylglycine or
L-arginine. Preferably, the pH buffer is a physiologically
acceptable buffer in a concentration in the range from about 3 mM
to about 20 mM, preferably from about 5 mM to about 15 mM. In a
preferred embodiment of this invention, the formulations of this
invention have a pH value is in the range from about 7.0 to about
8.0.
[0064] In a preferred embodiment of this invention, the formulation
is a neutral, aqueous soluble formulation containing a combination
of insulin aspart and insulin detemir in a concentration suitable
for infusion systems and, furthermore, essentially consisting of
phenolic preservatives (in a total concentration in the range from
about 30 mM to about 40 mM), glycerol as isotonicity agent (in a
concentration of about 0.17 M), dibasic sodium phosphate (in a
concentration in the range from about 5 mM to about 7 mM), zinc
ions corresponding to from about 2.5 to about 3.5 Zn.sup.2+/hexamer
insulin or from about 0.4 to about 0.6 Zn.sup.2+/monomer
insulin.
[0065] In a preferred embodiment of this invention, the formulation
of this invention has a content of non-dissolved material below
about 0.1%, preferably below 0.01% (weight per weight).
[0066] In a preferred embodiment of this invention, the stability
factor, when determined by the test described in example 1 above,
of the formulation of this invention is above about 2.5, preferably
above about 4.
[0067] In a preferred embodiment of this invention, a soluble
acylated insulin analogue is used in an amount in the range with
the lower limit of the molar ratio between the soluble acylated
insulin analogue and the monomeric insulin or human insulin being
7%, preferably 11%, more preferred 14%, and the upper limit of the
molar ratio between the soluble acylated insulin analogue and the
monomeric insulin or human insulin being 57%, preferably 41%, more
preferred 31%, preferably 24% and even more preferred 20%, of the
total amount of insulin to improve the physical stability in an
aqueous solution containing a monomeric insulin or human
insulin.
[0068] Administration of the formulations of this invention may be
via any route known to be effective by the physician of ordinary
skill. Parenteral and preferably subcutaneous and intraperitoneal
administration is preferred.
[0069] The amount of the formulation of this invention that is
administered to treat diabetes depends on a number of factors,
among which are included the patient's sex, weight, physical
activity, and age, diet of the patient, the underlying causes of
the condition or disease to be treated, the route of administration
and bioavailability, the persistence of the administered insulin or
insulin analogues in the body, the specific formulation used, the
potency of the insulin or insulin analogue used, a possible
combination with other drugs, the severity of the case of diabetes,
and the interval between dosages, if any interval. It is within the
skill of the ordinary physician to titrate the dose and infusion
rate and frequency of administration of the formulation of this
invention to achieve the desired result. It is recommended that the
daily dosage of the insulin components used in the formulations of
this invention be determined for each individual patient by those
skilled in the art in a similar way as for known insulin
compositions.
[0070] The mentioning herein of references is no admission that
they constitute prior art.
[0071] Herein, the word "comprise" is to be interpreted broadly
meaning "include", "contain" or "comprehend" (vide, for example,
EPO guidelines C 4.13).
[0072] This invention is further illustrated by the following
examples which, however, are not to be construed as limiting the
scope of protection. The features disclosed in the foregoing
description and in the following examples may, both separately and
in any combination thereof, be material for realizing this
invention in diverse forms thereof.
[0073] Stress Test
[0074] The insulin solutions prepared as described below were
subjected to a physical stress test.
[0075] Five samples of each insulin formulation were filled into
Penfill.RTM. 1.5 ml cartridges. After introduction of 100 .mu.L air
into each cartridge using a Hamilton.RTM. syringe, the samples were
subjected to the following physical stress test:
[0076] The cartridges were fixed to a rotator placed in an
incubator kept at a constant temperature of 37.degree.
C..+-.2.degree. C. and rotated 360.degree. at a frequency of 30 rpm
(rotations per minute) for four hours per day. The cartridges were
stored at a constant temperature of 37.degree. C..+-.2.degree. C.
between the rotation periods.
[0077] The opalescence of the cartridges was evaluated by visual
inspection at regular time intervals--once daily in the first week
and thereafter three times a week for up to 31 days. When
opalescence or precipitates occurs and is visible with the naked
eye, the sample is considered fibrillated. The number of days
without fibrillation is defined as the Fibrillation time. The
Stability Factor of a sample is calculated by dividing the
Fibrillation time of said sample with the Fibrillation time of a
specified reference sample tested in the same experiment.
[0078] The specified reference sample was insulin aspart 200 U/ml
which was prepared as follows:
[0079] A solution with the following composition was prepared:
Insulin aspart 200 U/ml (1200 nmol/ml), phenol 1.80 mg/ml (19 mM),
m-cresol 2.06 mg/ml (19 mM), glycerol 16 mg/ml (174 mM), dibasic
sodium phosphate dihydrate 1.25 mg/ml (7 mM), sodium chloride 0.58
mg/ml (10 mM), zinc chloride up to a total concentration of 39.2
.mu.g Zn.sup.2+/ml (3.0 Zn.sup.2+ per hexamer). Hydrochloric acid
and sodium hydroxide were used for dissolution of the insulin and
adjustment of the pH value to 7.40. Finally, the solution was
sterilized by filtration and filled into sterile Penfill.RTM.
cartridges 1.5 ml using aseptic technique.
[0080] Test Results
1 Fibrillation time (days) mean .+-. s.d. (n = 5) Stability Factor
Example 1 14 .+-. 12 3 aspart/detemir 98/2 Example 2 >31 >6
aspart/detemir 95/5 Example 3 >31 >6 aspart/detemir 90/10
Example 4 >31 >6 aspart/detemir 75/25 Reference 5 .+-. 3 1
aspart 100%
[0081] "s.d." is the standard deviation. As mentioned in the
examples below, for example, the figures 75/25 appearing after
"aspart/detemir" in this table indicates that the ratio between
aspart and detemir is 75:25 based upon insulin units.
[0082] Similar results as those stated in the above table were
obtained with corresponding formulations containing either 2.7
Zn.sup.2+/hexamer and a pH value of 7.60 or 3.0 Zn.sup.2+/hexamer
and a pH value of 7.60.
[0083] As appears from the above test results, the stability of
aspart against fibrillation is improved by adding detemir.
EXAMPLE 1
[0084] 200 U Insulin per ml Containing 98% (U/U) Insulin Aspart and
2% (U/U) Insulin Detemir. Molar Ratio Aspart/Detemir: 1:12.25
Corresponding to 7.55 mol-%.
[0085] A solution with the following composition was prepared:
Insulin aspart 196 U/ml (1176 nmol/ml), Insulin detemir 4 U/ml (96
nmol)ml, phenol 1.80 mg/ml (19 mM), m-cresol 2.06 mg/ml (19 mM),
glycerol 16 mg/ml (174 mM), dibasic sodium phosphate dihydrate 1.25
mg/ml (7 mM), sodium chloride 0.58 mg/ml (10 mM), zinc chloride and
zinc acetate up to a total concentration of 41.5 .mu.g Zn.sup.2+/ml
(3.0 Zn.sup.2+/hexamer). Hydrochloric acid and sodium hydroxide
were used for dissolution of the insulin and adjustment of the pH
value to 7.40. Finally, the solution was sterilized by filtration
and filled into sterile Penfill.RTM. cartridges 1.5 ml using
aseptic technique.
EXAMPLE 2
[0086] 200 U Insulin per ml Containing 95% (U/U) Insulin Aspart and
5% (U/U) Insulin Detemir Molar Ratio Aspart/Detemir: 1:4.75
Corresponding to 17.4 mol-%.
[0087] A solution with the following composition was prepared:
Insulin aspart 190 U/ml (1140 nmol/ml), Insulin detemir 10 U/ml
(240 nmol/ml), phenol 1.80 mg/ml (19 mM), m-cresol 2.06 mg/ml (19
mM), glycerol 16 mg/ml (174 mM), dibasic sodium phosphate dihydrate
1.25 mg/ml (7 mM), sodium chloride 0.58 mg/ml (10 mM), zinc
chloride and zinc acetate up to a total concentration of 45.1 .mu.g
Zn.sup.2+/ml (3.0 Zn.sup.2+/hexamer). Hydrochloric acid and sodium
hydroxide were used for dissolution of the insulin and adjustment
of the pH value to 7.40. Finally, the solution was sterilized by
filtration and filled into sterile Penfill.RTM. cartridges 1.5 ml
using aseptic technique.
EXAMPLE 3
[0088] 200 U Insulin per ml Containing 90% (U/U) Insulin Aspart and
10% (U/U) Insulin Detemir. Molar Ratio Aspart/Detemir: 1:2.25
Corresponding to 30.8 mol-%.
[0089] A solution with the following composition was prepared:
Insulin aspart 180 U/ml (1080 nmol/ml), Insulin detemir 20 U/ml
(480 nmol)ml, phenol 1.80 mg/ml (19 mM), m-cresol 2.06 mg/ml (19
mM), glycerol 16 mg/ml (174 mM), dibasic sodium phosphate dihydrate
1.25 mg/ml (7 mM), sodium chloride 0.58 mg/ml (10 mM), zinc
chloride and zinc acetate up to a total concentration of 51.0 .mu.g
Zn.sup.2+/ml (3.0 Zn.sup.2+/hexamer). Hydrochloric acid and sodium
hydroxide were used for dissolution of the insulin and adjustment
of the pH value to 7.40. Finally, the solution was sterilized by
filtration and filled into sterile Penfill.RTM. cartridges 1.5 ml
using aseptic technique.
EXAMPLE 4
[0090] 200 U Insulin per ml Containing 75% (U/U) Insulin Aspart and
25% (U/U) Insulin Detemir. Molar Ratio Aspart/Detemir: 1:0.75
Corresponding to 57.1 mol-%.
[0091] A solution with the following composition was prepared:
Insulin aspart 150 U/ml (900 nmol/ml), Insulin detemir 50 U/ml
(1200 nmol)ml, phenol 1.80 mg/ml (19 mM), m-cresol 2.06 mg/ml (19
mM), glycerol 16 mg/ml (174 mM), dibasic sodium phosphate dihydrate
1.25 mg/ml (7 mM), sodium chloride 0.58 mg/ml (10 mM), zinc
chloride and zinc acetate up to a total concentration of 68.6 .mu.g
Zn.sup.2+/ml (3.0 Zn.sup.2+/hexamer). Hydrochloric acid and sodium
hydroxide were used for dissolution of the insulin and adjustment
of the pH value to 7.40. Finally, the solution was sterilized by
filtration and filled into sterile Penfill.RTM. cartridges 1.5 ml
using aseptic technique.
EXAMPLE 5
[0092] 100 U Insulin per ml Containing 95% (U/U) Insulin Aspart and
5% (U/U) Insulin Detemir.
[0093] A solution with the following composition was prepared:
Insulin aspart 95 U/ml (570 nmol/ml), Insulin detemir 5 U/ml (120
nmol/ml), phenol 1.5 mg/ml (16 mM), m-cresol 1.7 mg/ml (16 mM),
glycerol 16 mg/ml (174 mM), dibasic sodium phosphate dihydrate 0.9
mg/ml (5 mM), sodium chloride 0.58 mg/ml (10 mM), zinc chloride and
zinc acetate up to a total concentration of 22.6 .mu.g Zn.sup.2+/ml
(3.0 Zn.sup.2+/hexamer). Hydrochloric acid and sodium hydroxide
were used for dissolution of the insulin and adjustment of the pH
value to 7.40. Finally, the solution was sterilized by filtration
and filled into sterile Penfill.RTM. cartridges 3 ml using aseptic
technique.
EXAMPLE 6
[0094] 100 U Insulin per ml Containing 90% (U/U) Insulin Aspart and
10% (U/U) Insulin Detemir.
[0095] A solution with the following composition was prepared:
Insulin aspart 90 U/ml (540 nmol/ml), Insulin detemir 10 U/ml (240
nmol/ml), phenol 1.5 mg/ml (16 mM), m-cresol 1.7 mg/ml (16 mM),
glycerol 16 mg/ml (174 mM), dibasic sodium phosphate dihydrate 0.9
mg/ml (5 mM), sodium chloride 0.58 mg/ml (10 mM), zinc chloride and
zinc acetate up to a total concentration of 25.5 .mu.g Zn.sup.2+/ml
(3.0 Zn.sup.2+/hexamer). Hydrochloric acid and sodium hydroxide
were used for dissolution of the insulin and adjustment of the pH
value to 7.40. Finally, the solution was sterilized by filtration
and filled into sterile Penfill.RTM. cartridges 3 ml using aseptic
technique.
EXAMPLE 7
[0096] 400 U Insulin per ml Containing 95% (U/U) Insulin Aspart and
5% (U/U) Insulin Detemir.
[0097] A solution with the following composition was prepared:
Insulin aspart 380 U/ml (2280 nmol/ml), Insulin detemir 20 U/ml
(480 nmol/ml), phenol 1.8 mg/ml (19 mM), m-cresol 2.1 mg/ml (19
mM), glycerol 16 mg/ml (174 mM), dibasic sodium phosphate dihydrate
0.9 mg/ml (5 mM), sodium chloride 1.2 mg/ml (20 mM), zinc chloride
and zinc acetate up to a total concentration of 90 .mu.g
Zn.sup.2+/ml (3.0 Zn.sup.2+/hexamer). Hydrochloric acid and sodium
hydroxide were used for dissolution of the insulin and adjustment
of the pH value to 7.40. Finally, the solution was sterilized by
filtration and filled into sterile Penfill.RTM. cartridges 1.5 ml
using aseptic technique.
* * * * *