U.S. patent application number 14/721889 was filed with the patent office on 2015-09-10 for injectable solution at ph 7 comprising at least one basal insulin whose pi is between 5.8 and 8.5.
This patent application is currently assigned to ADOCIA. The applicant listed for this patent is ADOCIA. Invention is credited to Alexandre GEISSLER, Gerard SOULA, Olivier SOULA, Jeff TONNAR.
Application Number | 20150250858 14/721889 |
Document ID | / |
Family ID | 47667929 |
Filed Date | 2015-09-10 |
United States Patent
Application |
20150250858 |
Kind Code |
A1 |
SOULA; Gerard ; et
al. |
September 10, 2015 |
INJECTABLE SOLUTION AT PH 7 COMPRISING AT LEAST ONE BASAL INSULIN
WHOSE PI IS BETWEEN 5.8 AND 8.5
Abstract
A composition in the form of an injectable aqueous solution, the
pH of which is between 6.0 and 8.0, includes at least a basal
insulin, the isoelectric point pI of which is between 5.8 and 8.5;
and a dextran substituted by radicals carrying carboxylate charges
and hydrophobic radicals. Single-dose formulations at a pH of
between 7 and 7.8 includes a basal insulin whose isoelectric point
is between 5.8 and 8.5 and a prandial insulin.
Inventors: |
SOULA; Gerard; (Meyzieu,
FR) ; SOULA; Olivier; (Meyzieu, FR) ; TONNAR;
Jeff; (Lyon, FR) ; GEISSLER; Alexandre; (Lyon,
FR) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
ADOCIA |
Lyon |
|
FR |
|
|
Assignee: |
ADOCIA
Lyon
FR
|
Family ID: |
47667929 |
Appl. No.: |
14/721889 |
Filed: |
May 26, 2015 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
13571026 |
Aug 9, 2012 |
9089476 |
|
|
14721889 |
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Current U.S.
Class: |
514/6.2 ;
514/5.9; 514/6.3; 514/6.4; 514/6.5 |
Current CPC
Class: |
A61K 38/28 20130101;
A61K 38/26 20130101; A61P 3/10 20180101; A61K 38/26 20130101; A61K
2300/00 20130101; A61K 2300/00 20130101; A61K 2300/00 20130101;
A61K 38/28 20130101; A61K 31/721 20130101; A61K 47/36 20130101;
A61K 9/08 20130101; A61K 9/0019 20130101; A61K 31/721 20130101;
A61P 43/00 20180101 |
International
Class: |
A61K 38/28 20060101
A61K038/28; A61K 38/26 20060101 A61K038/26; A61K 9/00 20060101
A61K009/00; A61K 47/36 20060101 A61K047/36; A61K 9/08 20060101
A61K009/08 |
Foreign Application Data
Date |
Code |
Application Number |
Aug 10, 2011 |
FR |
11/57291 |
Dec 23, 2011 |
FR |
11/62445 |
Claims
1. Composition in the form of an injectable aqueous solution, the
pH of which is between 6.0 and 8.0, comprising at least: a) a basal
insulin, the isoelectric point pI of which is between 5.8 and 8.5;
b) a dextran substituted by radicals carrying carboxylate charges
and hydrophobic radicals of formula I or of formula II:
##STR00043## in which: R is --OH or chosen from the group
consisting of the radicals: -(f-[A]-COOH).sub.n;
-(g-[B]-k-[D]).sub.m, D comprising at least one alkyl chain
comprising at least 8 carbon atoms; n represents the degree of
substitution of the glucoside units by -f-[A]-COOH and
0.1.ltoreq.n.ltoreq.2; m represents the degree of substitution of
the glucoside units by -g-[B]-k-[D] and 0<m.ltoreq.0.5; q
represents the degree of polymerization as glucoside units, that is
to say the mean number of glucoside units per polysaccharide chain,
and 3.ltoreq.q.ltoreq.50; -(f-[A]-COOH).sub.n: -A- is a linear or
branched radical comprising from 1 to 4 carbon atoms; the said -A-
radical: being bonded to a glucoside unit via a functional group f
chosen from the group consisting of ether, ester and carbamate
functional groups; -(g-[B]-k-[D]).sub.m: --B-- is a linear or
branched, at least divalent, radical comprising from 1 to 4 carbon
atoms; the said --B-- radical: being bonded to a glucoside unit via
a functional group g chosen from the group consisting of ether,
ester and carbamate functional groups; being bonded to a -D radical
via a functional group k; k chosen from the group consisting of
ester, amide and carbamate functional groups; the said -D radical:
being an --X(-l-Y).sub.p radical, X being an at least divalent
radical comprising from 1 to 12 atoms chosen from the group
consisting of C, N and O atoms, optionally carrying carboxyl or
amine functional groups and/or resulting from an amino acid, a
dialcohol, a diamine or a mono- or polyethylene glycol mono- or
diamine; Y being a linear or cyclic alkyl group, an alkylaryl or an
arylalkyl, of 8 to 30 carbon atoms; p.gtoreq.1 and l a functional
group chosen from the group consisting of ester, amide and
carbamate functional groups; f, g and k being identical or
different; the free acid functional groups being in the form of
salts of alkali metal cations chosen from the group consisting of
Na.sup.+ and K.sup.+; and, when p=1, if Y is a C.sub.8 to C.sub.14
alkyl, then q*m.gtoreq.2, if Y is a C.sub.15 alkyl, then
q*m.gtoreq.2; and if Y is a C.sub.16 to C.sub.20 alkyl, then
q*m.gtoreq.1; and, when p.gtoreq.2, if Y is a C.sub.8 to C.sub.9
alkyl, then q*m.gtoreq.2 and, if Y is a C.sub.10 to C.sub.16 alkyl,
then q*m.gtoreq.0.2; ##STR00044## in which: R is --OH or a
-(f-[A]-COOH).sub.n radical: -A- is a linear or branched radical
comprising from 1 to 4 carbon atoms; the said radical -A-: being
bonded to a glucoside unit via a functional group f chosen from the
group consisting of ether, ester or carbamate functional groups; n
represents the degree of substitution of the glucoside units by
-f-[A]-COOH and 0.1.ltoreq.n.ltoreq.2; R' is chosen from the group
consisting of the radicals: --C(O)NH-[E]-(o-[F]).sub.t;
--CH.sub.2N(L).sub.z-[E]-(o-[F]).sub.t; in which: z is a positive
integer equal to 1 or 2, L is chosen from the group consisting of:
--H and z is equal to 1, and/or -[A]-COOH and z is equal to 1 or 2,
if f is an ether functional group, --CO-[A]-COOH and z is equal to
1 if f is an ester functional group, and --CO--NH-[A]-COOH and z is
equal to 1 if f is a carbamate functional group;
-[E]-(o-[F]).sub.t: -E- is a linear or branched, at least divalent,
radical comprising from 1 to 8 carbon atoms and optionally
comprising heteroatoms, such as O, N or S; --F-- is a linear or
cyclic alkyl group, an alkylaryl or an arylalkyl, of 12 to 30
carbon atoms; o is a functional group chosen from the group
consisting of ether, ester, amide or carbamate functional groups; t
is a positive integer equal to 1 or 2; q represents the degree of
polymerization as glucoside units, that is to say the mean number
of glucoside units per polysaccharide chain, and
3.ltoreq.q.ltoreq.50; the free acid functional groups being in the
form of salts of alkali metal cations chosen from the group
consisting of Na.sup.+ and K.sup.+; when z=2, the nitrogen atom is
in the form of a quaternary ammonium.
2. Composition according to claim 1, wherein the dextran
substituted by radicals carrying carboxylate charges and
hydrophobic radicals is chosen from the dextrans of formula I.
3. Composition according to claim 1, wherein the dextran
substituted by radicals carrying carboxylate charges and
hydrophobic radicals is chosen from the dextrans of formula II.
4. Composition according to claim 1, wherein the dextran
substituted by radicals carrying carboxylate charges and
hydrophobic radicals is chosen from the dextrans of formula I in
which the -(f-[A]-COOH).sub.n radical is chosen from the group
consisting of the following sequences, f having the meaning given
above: ##STR00045##
5. Composition according to claim 1, wherein the dextran
substituted by radicals carrying carboxylate charges and
hydrophobic radicals is chosen from the dextrans of formula I in
which the -(g-[B]-k-[D]).sub.m radical is chosen from the group
consisting of the following sequences, g, k and D having the
meanings given above: ##STR00046##
6. Composition according to claim 1, wherein the dextran
substituted by radicals carrying carboxylate charges and
hydrophobic radicals is chosen from the dextrans of formula I in
which the -(g-[B]-k-[D]).sub.m radical is such that: --B-- is a
radical comprising one carbon atom; the said --B-- radical being
bonded to a glucoside unit via an ether functional group g, and X
is a radical resulting from an amino acid.
7. Composition according to claim 1, wherein the dextran
substituted by radicals carrying carboxylate charges and
hydrophobic radicals is chosen from the dextrans of formula I in
which the X radical is an at least divalent radical resulting from
an amino acid chosen from the group consisting of glycine, leucine,
phenylalanine, lysine, isoleucine, alanine, valine, aspartic acid
and glutamic acid.
8. Composition according to claim 1, wherein the dextran
substituted by radicals carrying carboxylate charges and
hydrophobic radicals is chosen from the dextrans of formula I in
which the Y group is chosen from the group consisting of a
hydrophobic alcohol, a hydrophobic acid, a sterol or a
tocopherol.
9. Composition according to claim 1, wherein the dextran
substituted by radicals carrying carboxylate charges and
hydrophobic radicals is chosen from the dextrans of formula I in
which the Y group is a sterol chosen from cholesterol
derivatives.
10. Composition according to claim 1, wherein the dextran
substituted by radicals carrying carboxylate charges and
hydrophobic radicals is chosen from the dextrans of formula II in
which the R' group is such that the -E- radical results from a
diamine.
11. Composition according to claim 1, wherein the dextran
substituted by radicals carrying carboxylate charges and
hydrophobic radicals is chosen from the dextrans of formula II in
which the R' group is such that the --F-- group results from a
cholesterol derivative.
12. Composition according to claim 1, wherein the dextran
substituted by radicals carrying carboxylate charges and
hydrophobic radicals is chosen from the group consisting of the
following dextrans of formula I: Sodium dextranmethylcarboxylate
modified by octyl glycinate, Sodium dextranmethylcarboxylate
modified by cetyl glycinate, Sodium dextranmethylcarboxylate
modified by octyl phenylalaninate, Sodium dextranmethylcarboxylate
modified by 3,7-dimethyl-1-octyl phenylalaninate, Sodium
dextranmethylcarboxylate modified by dioctyl aspartate, Sodium
dextranmethylcarboxylate modified by didecyl aspartate, Sodium
dextranmethylcarboxylate modified by dilauryl aspartate, Sodium
dextranmethylcarboxylate modified by N-(2-aminoethyl)dodecanamide,
Sodium dextransuccinate modified by lauryl glycinate, N-(sodium
methylcarboxylate) dextran carbamate modified by dioctyl aspartate,
Sodium dextranmethylcarboxylate modified by 2-(2-aminoethoxyl)ethyl
dodecanoate, Sodium dextranmethylcarboxylate modified by
2-(2-{2-[dodecanoylamino]ethoxy}ethoxy)ethylamine, Sodium
dextranmethylcarboxylate modified by
2-(2-{2-[hexadecanoylamino]ethoxy}ethoxy)ethylamine, Sodium
dextranmethylcarboxylate modified by cholesteryl leucinate, Sodium
dextranmethylcarboxylate modified by cholesteryl
1-ethylenediaminecarboxylate, and N-(sodium methylcarboxylate)
dextran carbamate modified by cholesteryl leucinate.
13. Composition according to claim 1, wherein the dextran
substituted by radicals carrying carboxylate charges and
hydrophobic radicals is chosen from the group consisting of the
dextrans of formula II and is: Sodium dextranmethylcarboxylate
modified by cholesteryl 1-ethylenediaminecarboxylate grafted by
reductive amination to the reducing chain end.
14. Composition according to claim 1, wherein the basal insulin
whose isoelectric point is between 5.8 and 8.5 is insulin
glargine.
15. Composition according to claim 1, wherein it comprises between
40 IU/ml and 500 IU/ml of basal insulin whose isoelectric point is
between 5.8 and 8.5.
16. Composition according to claim 1, wherein it additionally
comprises a prandial insulin.
17. Composition according to claim 1, wherein it comprises between
40 and 800 IU/ml of total insulin.
18. Composition according to claim 16, wherein it comprises
proportions, expressed as percentage, between the basal insulin
whose isoelectric point is between 5.8 and 8.5 and a prandial
insulin of 25/75, 30/70, 40/60, 50/50, 60/40, 70/30, 80/20 and
90/10.
19. Composition according to claim 1, wherein it additionally
comprises a GLP-1, a GLP-1 analogue or a GLP-1 derivative.
20. Composition according to claim 1, wherein it additionally
comprises zinc salts at a concentration of between 0 and 5000
.mu.M.
21. Composition according to claim 1, wherein it comprises buffers
chosen from the group consisting of Tris, citrates and phosphates
at concentrations of between 0 and 100 mM, preferably between 0 and
50 mM.
22. Composition according to claim 16, wherein the said prandial
insulin is chosen from the group formed by human insulin, insulin
glulisine, insulin lispro and insulin aspart.
23. Single-dose formulation comprising a composition according to
claim 1, at a pH of between 6.6 and 7.8, and a prandial
insulin.
24. Single-dose formulation comprising a composition according to
claim 1, at a pH of between 6.6 and 7.8, and a GLP-1, a GLP-1
derivative or a GLP-1 analogue.
25. Single-dose formulation according to claim 23, wherein the
prandial insulin is chosen from the group comprising of human
insulin.
26. Single-dose formulation according to claim 23, wherein the
prandial insulin is chosen from the group comprising of insulin
lispro, insulin glulisine and insulin aspart.
Description
[0001] This application is a continuation of U.S. application Ser.
No. 13/571,026 filed Aug. 9, 2012, which in turn claims the benefit
of U.S. Provisional Application No. 61/579,966 filed Dec. 23, 2011
and U.S. Provisional Application No. 61/522,031 filed Aug. 10,
2011, the entire disclosures of these prior applications being
incorporated herein by reference.
[0002] The invention relates to therapies by injection of
insulin(s) for treating diabetes.
[0003] Insulin therapy, or diabetes therapy by injecting insulin,
has made remarkable progress in recent years by virtue especially
of the development of novel insulins that offer correction of
patients' glycaemia, which enables better simulation of the
physiological activity of the pancreas.
[0004] To cover his daily insulin needs, a diabetic patient
currently has available, schematically, two types of insulin with
complementary actions: prandial insulins (or rapid-acting insulins)
and basal insulins (or slow-acting insulins).
[0005] Prandial insulins allow rapid management (metabolization
and/or storage) of the glucose provided by meals and snacks. The
patient must inject a prandial insulin before each food intake,
i.e. about 2 to 3 injections per day. The prandial insulins most
commonly used are: the recombinant human insulin, NovoLog.RTM.
(insulin aspart from Novo Nordisk), Humalog.RTM. (insulin lispro
from Eli Lilly) and Apidra.RTM. (insulin glulisine from
Sanofi-Aventis).
[0006] Basal insulins maintain the patient's glycaemic homeostasis,
outside periods of food intake. They act essentially by blocking
the endogenous production of glucose (hepatic glucose). The daily
dose of basal insulin generally corresponds to 40-50% of the total
daily insulin needs. Depending on the basal insulin used, this dose
is dispensed in 1 or 2 injections, regularly distributed throughout
the day. The basal insulins most commonly used are Levemir.RTM.
(insulin detemir from Novo Nordisk) and Lantus.RTM. (insulin
glargine from Sanofi-Aventis).
[0007] It will be noted, for the sake of completeness, that NPH
(NPH insulin, for Neutral Protamine Hagedorn; Humuline NPH.RTM.,
Insulatard.RTM.) is the oldest basal insulin. This formulation is
the result of a precipitation of human insulin (anionic at neutral
pH) with a cationic protein, protamine. These microcrystals are
dispersed in an aqueous suspension and dissolve slowly after
subcutaneous injection. This slow dissolution ensures sustained
release of the insulin. However, this release does not ensure a
constant concentration of insulin over time. The release profile is
bell-shaped and lasts only between 12 and 16 hours. It is thus
injected twice a day. This NPH basal insulin is much less efficient
than the modern basal insulins Levemir.RTM. and Lantus.RTM.. NPH is
an intermediate-acting basal insulin.
[0008] The principle of NPH changed with the appearance of rapid
insulin analogues to give "Premix" products that afford both rapid
action and intermediate action. NovoLog Mix.RTM. (Novo Nordisk) and
Humalog Mix.RTM. (Eli Lilly) are formulations comprising a rapid
insulin analogue, Novolog.RTM. and Humalog.RTM., partially
complexed with protamine. These formulations thus contain
microcrystals of insulin whose action is said to be intermediate,
and a proportion of insulin which has remained soluble, whose
action is rapid. These formulations do indeed offer the advantage
of a rapid insulin, but they also have the defect of NPH, i.e. a
duration of action limited to between 12 and 16 hours and a
bell-shaped release of insulin. However, these products enable the
patient to carry out a single injection of an intermediate-acting
basal insulin with a rapid-acting prandial insulin. In point of
fact, many patients are desirous to reduce their number of
injections.
[0009] The basal insulins currently marketed and currently under
clinical development may be categorized as a function of the
technical solution for obtaining sustained action, and two
approaches are used at the present time.
[0010] The first, that of insulin detemir, is the binding to
albumin in vivo. It is an analogue, soluble at pH 7, which
comprises a fatty acid (tetradecanoyl) side chain attached to
position B29 which, in vivo, enables this insulin to combine with
albumin. Its sustained action is mainly due to this affinity for
albumin after subcutaneous injection.
[0011] However, its pharmacokinetic profile does not make it
possible to cover a whole day, which means that it is usually used
in two injections per day.
[0012] Other basal insulins which are soluble at pH 7, such as
Degludec.RTM., are currently under development. Degludec.RTM. also
comprises a fatty acid side chain attached to insulin
(hexadecandoyl-.gamma.-L-Glu).
[0013] The second, that of insulin glargine, is the precipitation
at physiological pH. This is an analogue of human insulin obtained
by elongation of the C-terminal part of the B chain of human
insulin with two arginine residues, and by substitution of the
asparagine residue A21 with a glycine residue (U.S. Pat. No.
5,656,722). The addition of the two arginine residues was conceived
to adjust the pI (isoelectric point) of insulin glargine at
physiological pH, and thus make this insulin analogue insoluble in
physiological medium.
[0014] The substitution of A21 was conceived in order to make
insulin glargine stable at acidic pH and thus to be able to
formulate it in the form of an injectable solution at acidic pH.
During subcutaneous injection, the passage of insulin glargine from
an acidic pH (pH 4-4.5) to a physiological pH (neutral pH) brings
about its precipitation under the skin. The slow redissolution of
the insulin glargine microparticles ensures a slow and sustained
action.
[0015] The hypoglycaemic effect of insulin glargine is virtually
constant over a period of 24 hours, which allows the majority of
patients to limit themselves to a single injection per day.
[0016] Insulin glargine is nowadays considered as the best basal
insulin marketed.
[0017] However, the acidic pH of the formulations of basal
insulins, the isoelectric point of which is between 5.8 and 8.5, of
insulin glargine type, prevents any pharmaceutical combination with
other proteins and in particular prandial insulins, since the
latter are unstable at acidic pH.
[0018] However, no one has hitherto sought to dissolve these basal
insulins, the isoelectric point of which is between 5.8 and 8.5, of
insulin glargine type, at neutral pH, while maintaining a
difference in solubility between the in vitro medium (containing
it) and the in vivo medium (under the skin), independently of the
pH.
[0019] Specifically, the operating principle of basal insulins, of
insulin glargine type, outlined above, namely that they are soluble
at acidic pH and precipitate at physiological pH, dissuades a
person skilled in the art from any solution in which insulin of
insulin glargine type would be dissolved at pH 6-8 while
maintaining its essential property, which is that of precipitating
in subcutaneous medium.
[0020] Furthermore, the impossibility of formulating a prandial
insulin, at acidic pH, arises from the fact that a prandial insulin
undergoes, under these conditions, a deamidation side reaction in
position A21; this does not make it possible to satisfy the
requirement of the US Pharmacopeia, namely less than 5% of
by-product after 4 weeks at 30.degree. C.
[0021] Furthermore, this acidic pH of formulations of basal
insulins, the isoelectric point of which is between 5.8 and 8.5, of
insulin glargine type, even prevents any extemporaneous combination
with prandial insulins at neutral pH.
[0022] A recent clinical study, presented at the 69th Scientific
Sessions of the American Diabetes Association, New Orleans, La.,
5-9 Jun. 2009, verified this limitation of use of insulin glargine.
A dose of insulin glargine and a dose of prandial insulin (in the
case in point, insulin lispro) were mixed together just before
injection (E. Cengiz et al., 2010, Diabetes Care, 33(5), 1009-12).
This experiment made it possible to demonstrate a significant delay
in the pharmacokinetic and pharmacodynamic profiles of prandial
insulin, which may give rise to postprandial hyperglycaemia and to
nocturnal hypoglycaemia. This study indeed confirms the
incompatibility of insulin glargine with the currently marketed
rapid-acting insulins.
[0023] Moreover, the instructions for use for Lantus.RTM., the
commercial product based on insulin glargine from the company
Sanofi-Aventis, explicitly indicates to users that they should not
mix it with a solution of prandial insulin, whichever it may be, on
account of the serious risk of modifying the pharmacokinetics and
pharmacodynamics of the insulin glargine and/or of the mixed
prandial insulin.
[0024] However, from a therapeutic point of view, clinical studies
made public during the 70th annual scientific sessions of the
American Diabetes Association (ADA) of 2010, abstract 2163-PO and
abstract number 0001-LB, in particular those conducted by the
company Sanofi-Aventis, showed that treatments which combine
Lantus.RTM., insulin glargine, and a prandial insulin are much more
efficient than treatments based on products of the "Premix",
Novolog Mix.RTM. or Humalog Mix.RTM. type.
[0025] As regards combinations of insulin glargine and rapid
insulin, the company Biodel has notably described, in patent
application U.S. Pat. No. 7,718,609, compositions comprising a
basal insulin and a prandial insulin at a pH of between 3.0 and 4.2
in the presence of a chelating agent and polyacids. This patent
teaches how to make a prandial insulin compatible at acidic pH in
the presence of an insulin of insulin glargine type. It does not
teach how to prepare a combination of insulin of insulin glargine
type and of a prandial insulin at neutral pH.
[0026] From the analysis of the compositions described in the
literature and the patents, it appears that the insolubility at pH
7 of basal insulins of the insulin glargine type is a prerequisite
for having its slow action. As a result, all the solutions proposed
for combining them with other products, such as prandial insulins,
are based on tests of dissolution or stabilization of prandial
insulins at acidic pH; see, for example, WO 2007/121256 and WO
2009/021955.
[0027] Surprisingly, the compositions according to the invention
make it possible to dissolve, at pH 7, a basal insulin whose
isoelectric point is between 5.8 and 8.5.
[0028] Surprisingly, the compositions according to the invention
allow maintenance of the duration of the hypoglycaemic activity of
the basal insulin whose isoelectric point is between 5.8 and 8.5,
despite its dissolution at pH 7 before injection. This noteworthy
property arises from the fact that the insulin of insulin glargine
type dissolved at pH 7 in the composition of the invention
precipitates in subcutaneous medium by means of a change of
composition of the medium. The factor triggering the precipitation
of the insulin of insulin glargine type is no longer the pH
modification, but a modification of composition of the environment
during the passage of the pharmaceutical composition into the
physiological medium.
[0029] By solving this problem of solubility at pH 7, the present
invention makes it possible: [0030] to propose an injectable
composition, intended for treating diabetes, comprising a basal
insulin whose isoelectric point is between 5.8 and 8.5, in the form
of a homogeneous solution at pH 7, while retaining its biological
activity and its action profile; [0031] to propose a composition in
the form of a formulation comprising a combination of a basal
insulin whose isoelectric point is between 5.8 and 8.5 and a
prandial insulin without modification of the activity profile of
the prandial insulin which is soluble at pH 6-8 and unstable at
acidic pH, while maintaining the basal action profile intrinsic to
the basal insulin; [0032] to propose an injectable composition,
intended for treating diabetes, additionally comprising a
combination of a basal insulin whose isoelectric point is between
5.8 and 8.5 and a derivative or an analogue of a gastrointestinal
hormone such as GLP-1 or "glucagon-like peptide-1"; [0033] for
patients to reduce their number of injections; [0034] for the said
compositions to satisfy the requirements of the American
Pharmacopeia and European Pharmacopoeia.
[0035] Surprisingly, in insulin combinations of insulin glargine
type with a prandial insulin, which are subject-matters of the
invention, the rapid action of the prandial insulin is preserved
despite the precipitation of the insulin of insulin glargine type
in subcutaneous medium.
[0036] The invention relates to a composition in the form of an
injectable aqueous solution, the pH of which is between 6.0 and
8.0, comprising at least: [0037] a) a basal insulin, the
isoelectric point pI of which is between 5.8 and 8.5; [0038] b) a
dextran substituted by radicals carrying carboxylate charges and
hydrophobic radicals of formula I or of formula II:
##STR00001##
[0039] in which: [0040] R is --OH or chosen from the group
consisting of the radicals: [0041] -(f-[A]-COOH).sub.n; [0042]
-(g-[B]-k-[D]).sub.m, D comprising at least one alkyl chain
comprising at least 8 carbon atoms; [0043] n represents the degree
of substitution of the glucoside units by -f-[A]-COOH and
0.1.ltoreq.n.ltoreq.2; [0044] m represents the degree of
substitution of the glucoside units by -g-[B]-k-[D] and
0<m.ltoreq.0.5; [0045] q represents the degree of polymerization
as glucoside units, that is to say the mean number of glucoside
units per polysaccharide chain, and 3.ltoreq.q.ltoreq.50; [0046]
-(f-[A]-COOH).sub.n: [0047] -A- is a linear or branched radical
comprising from 1 to 4 carbon atoms; the said -A- radical: [0048]
being bonded to a glucoside unit via a functional group f chosen
from the group consisting of ether, ester and carbamate functional
groups; [0049] -(g-[B]-k-[D]).sub.m: [0050] --B-- is a linear or
branched, at least divalent, radical comprising from 1 to 4 carbon
atoms; the said --B-- radical: [0051] being bonded to a glucoside
unit via a functional group g chosen from the group consisting of
ether, ester and carbamate functional groups; [0052] being bonded
to a -D radical via a functional group k; k chosen from the group
consisting of ester, amide and carbamate functional groups; the
said -D radical: [0053] being an --X(-l-Y).sub.p radical, X being
an at least divalent radical comprising from 1 to 12 atoms chosen
from the group consisting of C, N and O atoms, optionally carrying
carboxyl or amine functional groups and/or resulting from an amino
acid, a dialcohol, a diamine or a mono- or polyethylene glycol
mono- or diamine; Y being a linear or cyclic alkyl group, an
alkylaryl or an arylalkyl, of 8 to 30 carbon atoms, optionally
substituted by one or more C.sub.1 to C.sub.3 alkyl groups;
p.gtoreq.1 and l a functional group chosen from the group
consisting of ester, amide and carbamate functional groups; [0054]
f, g and k being identical or different; [0055] the free acid
functional groups being in the form of salts of alkali metal
cations chosen from the group consisting of Na.sup.+ and K.sup.+;
[0056] and, when p=1, if Y is a C.sub.8 to C.sub.14 alkyl, then
q*m.gtoreq.2, if Y is a C.sub.15 alkyl, then q*m.gtoreq.2; and if Y
is a C.sub.16 to C.sub.20 alkyl, then q*m.gtoreq.1; [0057] and,
when p.gtoreq.2, if Y is a C.sub.8 to C.sub.9 alkyl, then
q*m.gtoreq.2 and, if Y is a C.sub.10 to C.sub.16 alkyl, then
q*m.gtoreq.0.2;
##STR00002##
[0057] in which: [0058] R is --OH or a -(f-[A]-COOH).sub.n radical:
[0059] -A- is a linear or branched radical comprising from 1 to 4
carbon atoms; the said radical -A-: [0060] being bonded to a
glucoside unit via a functional group f chosen from the group
consisting of ether, ester or carbamate functional groups; [0061] n
represents the degree of substitution of the glucoside units by
-f-[A]-COOH and 0.1.ltoreq.n.ltoreq.2; [0062] R' is chosen from the
group consisting of the radicals: [0063]
--C(O)NH-[E]-(o-[F]).sub.t; [0064]
--CH.sub.2N(L).sub.z-[E]-(o-[F]).sub.t; [0065] in which: [0066] z
is a positive integer equal to 1 or 2, [0067] L is chosen from the
group consisting of: [0068] --H and z is equal to 1, and/or [0069]
-[A]-COOH and z is equal to 1 or 2, if f is an ether functional
group, [0070] --CO-[A]-COOH and z is equal to 1 if f is an ester
functional group, and [0071] --CO--NH-[A]-COOH and z is equal to 1
if f is a carbamate functional group; [0072] -[E]-(o-[F]).sub.t:
[0073] -E- is a linear or branched, at least divalent, radical
comprising from 1 to 8 carbon atoms and optionally comprising
heteroatoms, such as O, N or S; [0074] --F-- is a linear or cyclic
alkyl group, an alkylaryl or an arylalkyl, of 12 to 30 carbon
atoms, optionally substituted by one or more C.sub.1 to C.sub.3
alkyl groups; [0075] o is a functional group chosen from the group
consisting of ether, ester, amide or carbamate functional groups;
[0076] t is a positive integer equal to 1 or 2; [0077] q represents
the degree of polymerization as glucoside units, that is to say the
mean number of glucoside units per polysaccharide chain, and
3.ltoreq.q.ltoreq.50; [0078] the free acid functional groups being
in the form of salts of alkali metal cations chosen from the group
consisting of Na.sup.+ and K.sup.+; [0079] when z=2, the nitrogen
atom is in the form of a quaternary ammonium.
[0080] In one embodiment, when p=1, if Y is a C.sub.21 to C.sub.30
group, then q*m.gtoreq.1.
[0081] In one embodiment, when p=1, if Y is a C.sub.21 to C.sub.30
group, then q*m.gtoreq.0.1.
[0082] In one embodiment, the -(f-[A]-COOH).sub.n radical is such
that: [0083] -A- is a radical comprising one carbon atom; the said
-A- radical being bonded to a glucoside unit via a ether functional
group f.
[0084] In one embodiment, the -(g-[B]-k-[D]).sub.m radical is such
that: [0085] --B-- is a radical comprising one carbon atom; the
said --B-- radical being bonded to a glucoside unit via an ether
functional group g, and [0086] X is a radical resulting from an
amino acid.
[0087] In one embodiment, the -(f-[A]-COOH).sub.n radical is such
that: [0088] -A- is a radical comprising one carbon atom; the said
-A- radical being bonded to a glucoside unit via an ether
functional group f, and [0089] the -(g-[B]-k-[D]).sub.m radical is
such that: [0090] --B-- is a radical comprising one carbon atom;
the said --B-- radical being bonded to a glucoside unit via an
ether functional group g, and [0091] X is a radical resulting from
an amino acid, [0092] k is an amide functional group.
[0093] In one embodiment, the dextran substituted by radicals
carrying carboxylate charges and hydrophobic radicals is of formula
III:
##STR00003##
[0094] in which: [0095] R is --OH or chosen from the group
consisting of the radicals: [0096] -(f-[A]-COOH).sub.n; [0097]
-(g-[B]-k-[D]).sub.m, D comprising at least one alkyl chain
comprising at least 8 carbon atoms; [0098] n represents the degree
of substitution of the glucoside units by -f-[A]-COOH and
0.1.ltoreq.n.ltoreq.2; [0099] m represents the degree of
substitution of the glucoside units by -g-[B]-k-[D] and
0<m.ltoreq.0.5; [0100] q represents the degree of polymerization
as glucoside units, that is to say the mean number of glucoside
units per polysaccharide chain, and 3.ltoreq.q.ltoreq.50; [0101]
-(f-[A]-COOH).sub.n: [0102] -A- is a linear or branched radical
comprising from 1 to 4 carbon atoms; the said -A- radical: [0103]
being bonded to a glucoside unit via a functional group f chosen
from the group consisting of ether, ester and carbamate functional
groups; [0104] -(g-[B]-k-[D]).sub.m: [0105] --B-- is a linear or
branched, at least divalent, radical comprising from 1 to 4 carbon
atoms; the said --B-- radical: [0106] being bonded to a glucoside
unit via a functional group g chosen from the group consisting of
ether, ester and carbamate functional groups; [0107] being bonded
to a -D radical via a functional group k; k chosen from the group
consisting of ester, amide and carbamate functional groups; the
said -D radical: [0108] being an --X(-l-Y).sub.p radical, X being
an at least divalent radical comprising from 1 to 12 atoms chosen
from the group consisting of C, N and O atoms, optionally carrying
carboxyl or amine functional groups and/or resulting from an amino
acid, a dialcohol, a diamine or a mono- or polyethylene glycol
mono- or diamine; Y being a linear or cyclic alkyl group, an
alkylaryl or an arylalkyl, of 8 to 20 carbon atoms, optionally
substituted by one or more C.sub.1 to C.sub.3 alkyl groups;
p.gtoreq.1 and l a functional group chosen from the group
consisting of ester, amide and carbamate functional groups; [0109]
f, g and k being identical or different; [0110] the free acid
functional groups being in the form of salts of alkali metal
cations chosen from the group consisting of Na.sup.+ and K.sup.+;
[0111] and, when p=1, if Y is a C.sub.8 to C.sub.14 alkyl, then
q*m.gtoreq.2, if Y is a C.sub.15 alkyl, then q*m.gtoreq.2; and if Y
is a C.sub.16 to C.sub.20 alkyl, then q*m.gtoreq.1; [0112] and,
when p.gtoreq.2, if Y is a C.sub.8 to C.sub.11 alkyl, then
q*m.gtoreq.2 and, if Y is a C.sub.12 to C.sub.16 alkyl, then
q*m.gtoreq.0.3.
[0113] In one embodiment, the dextran substituted by radicals
carrying carboxylate charges and hydrophobic radicals is of formula
IV:
##STR00004##
[0114] in which: [0115] R is --OH or chosen from the group
consisting of the radicals: [0116] -(f-[A]-COOH).sub.n; [0117]
-(g-[B]-k-[D]).sub.m, D comprising at least one alkyl chain
comprising at least 8 carbon atoms; [0118] n represents the degree
of substitution of the hydroxyl --OH functional groups by
-f-[A]-COOH per glucoside unit; and 0.1.ltoreq.n.ltoreq.2; [0119] m
represents the degree of substitution of the hydroxyl --OH
functional groups by -g-[B]-k-[D] per glucoside unit; and
0<m.ltoreq.0.5; [0120] q represents the degree of polymerization
as glucoside units, that is to say the mean number of glucoside
units per polysaccharide chain, and 3.ltoreq.q.ltoreq.50; [0121]
-(f-[A]-COOH).sub.n: [0122] -A- is a linear or branched radical
comprising from 1 to 4 carbon atoms; the said -A- radical: [0123]
being bonded to a glucoside unit via a functional group f chosen
from the group consisting of ether, ester and carbamate functional
groups; [0124] -(g-[B]-k-[D]).sub.m: [0125] --B-- is a linear or
branched, at least divalent, radical comprising from 1 to 4 carbon
atoms; the said --B-- radical: [0126] being bonded to a glucoside
unit via a functional group g chosen from the group consisting of
ether, ester and carbamate functional groups; [0127] being bonded
to a -D radical via a functional group k; k chosen from the group
consisting of ester, amide and carbamate functional groups; the
said -D radical: [0128] being an --X(-l-Y).sub.p radical, X being
an at least divalent radical comprising from 1 to 12 atoms chosen
from the group consisting of C, N and O atoms, optionally carrying
carboxyl or amine functional groups and/or resulting from an amino
acid, a dialcohol, a diamine or a mono- or polyethylene glycol
mono- or diamine; Y being a linear or cyclic alkyl group, an
alkylaryl or an arylalkyl, of 8 to 30 carbon atoms, optionally
substituted by one or more C.sub.1 to C.sub.3 alkyl groups;
p.gtoreq.1 and l a functional group chosen from the group
consisting of ester, amide and carbamate functional groups; [0129]
f, g and k being identical or different; [0130] the free acid
functional groups being in the form of salts of alkali metal
cations chosen from the group consisting of Na.sup.+ and K.sup.+;
[0131] and, when p=1, if Y is a C.sub.8 to C.sub.14 alkyl, then
q*m.gtoreq.2, if Y is a C.sub.15 alkyl, then q*m.gtoreq.2; and if Y
is a C.sub.16 to C.sub.30 alkyl, then q*m.gtoreq.1; [0132] and,
when p.gtoreq.2, if Y is a C.sub.8 to C.sub.9 alkyl, then
q*m.gtoreq.2 and, if Y is a C.sub.10 to C.sub.16 alkyl, then
q*m.gtoreq.0.2.
[0133] The structure drawn corresponds to the representation
commonly used to represent dextran, which is a polysaccharide
predominantly composed of (1,6) linkages between glucoside units,
which is the representation adopted. Dextran also comprises (1,3)
linkages at approximately 5% in general, which are deliberately not
represented but which are, of course, included within the scope of
the invention.
[0134] The term "basal insulin" whose isoelectric point is between
5.8 and 8.5 is understood to mean an insulin which is insoluble at
pH 7 and which has a duration of action of between 8 and 24 hours
or more in the standard models of diabetes.
[0135] These basal insulins whose isoelectric point is between 5.8
and 8.5 are recombinant insulins, the primary structure of which
has been modified mainly by introduction of basic amino acids, such
as arginine or lysine. They are described, for example, in the
following patents, patent applications or publications: WO
2003/053339, WO 2004/096854, U.S. Pat. No. 5,656,722 and U.S. Pat.
No. 6,100,376.
[0136] In one embodiment, the basal insulin whose isoelectric point
is between 5.8 and 8.5 is insulin glargine.
[0137] In one embodiment, the compositions according to the
invention comprise 100 IU/ml (i.e., approximately 3.6 mg/ml) of
basal insulin whose isoelectric point is between 5.8 and 8.5.
[0138] In one embodiment, the compositions according to the
invention comprise 40 IU/ml of basal insulin whose isoelectric
point is between 5.8 and 8.5.
[0139] In one embodiment, the compositions according to the
invention comprise 200 IU/ml of basal insulin whose isoelectric
point is between 5.8 and 8.5.
[0140] In one embodiment, the compositions according to the
invention comprise 300 IU/ml of basal insulin whose isoelectric
point is between 5.8 and 8.5.
[0141] In one embodiment, the compositions according to the
invention comprise 400 IU/ml of basal insulin whose isoelectric
point is between 5.8 and 8.5.
[0142] In one embodiment, the compositions according to the
invention comprise 500 IU/ml of basal insulin whose isoelectric
point is between 5.8 and 8.5.
[0143] In one embodiment, the ratio by weight of the basal insulin
whose isoelectric point is between 5.8 and 8.5 to the substituted
dextran, i.e. substituted dextran/basal insulin, is between 0.2 and
5.
[0144] In one embodiment, the ratio by weight of the basal insulin
whose isoelectric point is between 5.8 and 8.5 to the substituted
dextran, i.e. substituted dextran/basal insulin, is between 0.2 and
4.
[0145] In one embodiment, the ratio by weight of the basal insulin
whose isoelectric point is between 5.8 and 8.5 to the substituted
dextran, i.e. substituted dextran/basal insulin, is between 0.2 and
3.
[0146] In one embodiment, the ratio by weight of the basal insulin
whose isoelectric point is between 5.8 and 8.5 to the substituted
dextran, i.e. substituted dextran/basal insulin, is between 0.5 and
3.
[0147] In one embodiment, the ratio by weight of the basal insulin
whose isoelectric point is between 5.8 and 8.5 to the substituted
dextran, i.e. substituted dextran/basal insulin, is between 0.8 and
3.
[0148] In one embodiment, the ratio by weight of the basal insulin
whose isoelectric point is between 5.8 and 8.5 to the substituted
dextran, i.e. substituted dextran/basal insulin, is between 1 and
3.
[0149] In one embodiment, the concentration of substituted dextran
is between 1 and 100 mg/ml.
[0150] In one embodiment, the concentration of substituted dextran
is between 1 and 80 mg/ml.
[0151] In one embodiment, the concentration of substituted dextran
is between 1 and 60 mg/ml.
[0152] In one embodiment, the concentration of substituted dextran
is between 1 and 50 mg/ml.
[0153] In one embodiment, the concentration of substituted dextran
is between 1 and 30 mg/ml.
[0154] In one embodiment, the concentration of substituted dextran
is between 1 and 20 mg/ml.
[0155] In one embodiment, the concentration of substituted dextran
is between 1 and 10 mg/ml.
[0156] In one embodiment, the concentration of polysaccharide is
between 5 and 20 mg/ml.
[0157] In one embodiment, the concentration of polysaccharide is
between 5 and 10 mg/ml.
[0158] In one embodiment, the compositions according to the
invention additionally comprise a prandial insulin. Prandial
insulins are soluble at pH 7.
[0159] The term "prandial insulin" is understood to mean a "rapid"
or "regular" insulin.
[0160] "Rapid" prandial insulins are insulins which must meet the
needs brought about by the ingestion of proteins and carbohydrates
during a meal; they have to act in less than 30 minutes.
[0161] In one embodiment, "regular" prandial insulins are chosen
from the group consisting of Humulin.RTM. (human insulin) and
Novolin.RTM. (human insulin).
[0162] "Fast-acting" prandial insulins are insulins which are
obtained by recombination and which are modified in order to reduce
their action time.
[0163] In one embodiment, "fast-acting" prandial insulins are
chosen from the group consisting of insulin lispro (Humalog.RTM.),
insulin glulisine (Apidra.RTM.) and insulin aspart
(NovoLog.RTM.).
[0164] In one embodiment, the compositions according to the
invention comprise, in total, 100 IU/ml (i.e., approximately 3.6
mg/ml) of insulin with a combination of prandial insulin and basal
insulin whose isoelectric point is between 5.8 and 8.5.
[0165] In one embodiment, the compositions according to the
invention comprise, in total, 40 IU/ml of insulin with a
combination of prandial insulin and basal insulin whose isoelectric
point is between 5.8 and 8.5.
[0166] In one embodiment, the compositions according to the
invention comprise, in total, 200 IU/ml of insulin with a
combination of prandial insulin and basal insulin whose isoelectric
point is between 5.8 and 8.5.
[0167] In one embodiment, the compositions according to the
invention comprise, in total, 300 IU/ml of insulin with a
combination of prandial insulin and basal insulin whose isoelectric
point is between 5.8 and 8.5.
[0168] In one embodiment, the compositions according to the
invention comprise, in total, 400 IU/ml of insulin with a
combination of prandial insulin and basal insulin whose isoelectric
point is between 5.8 and 8.5.
[0169] In one embodiment, the compositions according to the
invention comprise, in total, 500 IU/ml of insulin with a
combination of prandial insulin and basal insulin whose isoelectric
point is between 5.8 and 8.5.
[0170] In one embodiment, the compositions according to the
invention comprise, in total, 600 IU/ml of insulin with a
combination of prandial insulin and basal insulin whose isoelectric
point is between 5.8 and 8.5.
[0171] In one embodiment, the compositions according to the
invention comprise, in total, 700 IU/ml of insulin with a
combination of prandial insulin and basal insulin whose isoelectric
point is between 5.8 and 8.5.
[0172] In one embodiment, the compositions according to the
invention comprise, in total, 800 IU/ml of insulin with a
combination of prandial insulin and basal insulin whose isoelectric
point is between 5.8 and 8.5.
[0173] The proportions between the basal insulin whose isoelectric
point is between 5.8 and 8.5 and the prandial insulin, expressed as
percentage with respect to the total amount of insulin, are, for
example, 25/75, 30/70, 40/60, 50/50, 60/40, 70/30, 80/20 and 90/10
for the formulations as described above from 40 to 800 IU/ml.
However, any other proportion can be produced.
[0174] For a formulation comprising 100 IU/ml as total insulin, the
proportions between the basal insulin whose isoelectric point is
between 5.8 and 8.5 and the prandial insulin are, for example, in
IU/ml, 25/75, 30/70, 40/60, 50/50, 60/40, 70/30, 80/20 or 90/10.
However, any other proportion can be produced.
[0175] In one embodiment, the composition according to the
invention additionally comprises a GLP-1, a GLP-1 analogue or a
GLP-1 derivative.
[0176] In one embodiment, the GLP-1 analogues or derivatives are
chosen from the group consisting of exenatide or Byetta.RTM.,
developed by Eli Lilly & Co and Amylin Pharmaceuticals,
liraglutide or Victoza.RTM. developed by Novo Nordisk, or
lixisenatide or Lyxumia.RTM. developed by Sanofi, their analogues
or derivatives and their pharmaceutically acceptable salts.
[0177] In one embodiment, the GLP-1 analogue or derivative is
exenatide or Byetta.RTM., its analogues or derivatives and their
pharmaceutically acceptable salts.
[0178] In one embodiment, the GLP-1 analogue or derivative is
liraglutide or Victoza.RTM., its analogues or derivatives and their
pharmaceutically acceptable salts.
[0179] In one embodiment, the GLP-1 analogue or derivative is
lixisenatide or Lyxumia.RTM., its analogues or derivatives and
their pharmaceutically acceptable salts.
[0180] The term "analogue" is understood to mean, when it is used
with reference to a peptide or a protein, a peptide or a protein in
which one or more constituent amino acid residues have been
replaced by other amino acid residues and/or from which one or more
constituent amino acid residues have been deleted and/or to which
one or more constituent amino acid residues have been added. The
percentage of homology accepted for the present definition of an
analogue is 50%.
[0181] The term "derivative" is understood to mean, when it is used
with reference to a peptide or protein, a peptide or a protein or
an analogue chemically modified by a substituent which is not
present in the reference peptide or protein or analogue, that is to
stay a peptide or a protein which has been modified by creation of
covalent bonds, in order to introduce substituents.
[0182] In one embodiment, the concentration of GLP-1, of GLP-1
analogue or of GLP-1 derivative is within a range from 0.01 to 10
mg/ml.
[0183] In one embodiment, the concentration of exenatide, its
analogues or derivatives and their pharmaceutically acceptable
salts is within a range from 0.05 to 0.5 mg/ml.
[0184] In one embodiment, the concentration of liraglutide, its
analogues or derivatives and their pharmaceutically acceptable
salts is within a range from 1 to 10 mg/ml.
[0185] In one embodiment, the concentration of lixisenatide, its
analogues or derivatives and their pharmaceutically acceptable
salts is within a range from 0.01 to 1 mg/ml.
[0186] In one embodiment, the compositions according to the
invention are produced by mixing commercial solutions of basal
insulin whose isoelectric point is between 5.8 and 8.5 and
commercial solutions of GLP-1, of GLP-1 analogue or of GLP-1
derivative in ratios by volume within a range from 10/90 to
90/10.
[0187] In one embodiment, the composition according to the
invention comprises a daily dose of basal insulin and a daily dose
of GLP-1, GLP-1 analogue or GLP-1 derivative.
[0188] In one embodiment, the compositions according to the
invention comprise 500 IU/ml of basal insulin whose isoelectric
point is between 5.8 and 8.5 and from 0.05 to 0.5 mg/ml of
exenatide.
[0189] In one embodiment, the compositions according to the
invention comprise 500 IU/ml of basal insulin whose isoelectric
point is between 5.8 and 8.5 and from 1 to 10 mg/ml of
liraglutide.
[0190] In one embodiment, the compositions according to the
invention comprise 500 IU/ml of basal insulin whose isoelectric
point is between 5.8 and 8.5 and from 0.05 to 0.5 mg/ml of
lixisenatide.
[0191] In one embodiment, the compositions according to the
invention comprise 100 IU/ml of basal insulin whose isoelectric
point is between 5.8 and 8.5 and from 0.05 to 0.5 mg/ml of
exenatide.
[0192] In one embodiment, the compositions according to the
invention comprise 100 IU/ml of basal insulin whose isoelectric
point is between 5.8 and 8.5 and from 1 to 10 mg/ml of
liraglutide.
[0193] In one embodiment, the compositions according to the
invention comprise 100 IU/ml of basal insulin whose isoelectric
point is between 5.8 and 8.5 and from 0.05 to 0.5 mg/ml of
lixisenatide.
[0194] In one embodiment, the compositions according to the
invention comprise 40 IU/ml of basal insulin whose isoelectric
point is between 5.8 and 8.5 and from 0.05 a 0.5 mg/ml of
exenatide.
[0195] In one embodiment, the compositions according to the
invention comprise 40 IU/ml of basal insulin whose isoelectric
point is between 5.8 and 8.5 and from 1 to 10 mg/ml of
liraglutide.
[0196] In one embodiment, the compositions according to the
invention comprise 40 IU/ml of basal insulin whose isoelectric
point is between 5.8 and 8.5 and from 0.05 a 0.5 mg/ml of
lixisenatide.
[0197] In one embodiment, the compositions according to the
invention comprise 200 IU/ml of basal insulin whose isoelectric
point is between 5.8 and 8.5 and from 0.05 to 0.5 mg/ml of
exenatide.
[0198] In one embodiment, the compositions according to the
invention comprise 200 IU/ml of basal insulin whose isoelectric
point is between 5.8 and 8.5 and from 1 to 10 mg/ml of
liraglutide.
[0199] In one embodiment, the compositions according to the
invention comprise 200 IU/ml of basal insulin whose isoelectric
point is between 5.8 and 8.5 and from 0.05 to 0.5 mg/ml of
lixisenatide.
[0200] In one embodiment, the compositions according to the
invention additionally comprise zinc salts at a concentration of
between 0 and 5000 .mu.M.
[0201] In one embodiment, the compositions according to the
invention additionally comprise zinc salts at a concentration of
between 50 and 4000 .mu.M.
[0202] In one embodiment, the compositions according to the
invention additionally comprise zinc salts at a concentration of
between 200 and 3000 .mu.M.
[0203] In one embodiment, the compositions according to the
invention additionally comprise zinc salts at a concentration of
between 0 and 1000 .mu.M.
[0204] In one embodiment, the compositions according to the
invention additionally comprise zinc salts at a concentration of
between 20 and 600 .mu.M.
[0205] In one embodiment, the compositions according to the
invention additionally comprise zinc salts at a concentration of
between 50 and 500 .mu.M.
[0206] In one embodiment, the compositions according to the
invention comprise buffers chosen from the group consisting of
Tris, citrates and phosphates at concentrations of between 0 and
100 mM, preferably between 0 and 50 mM or between 15 and 50 mM.
[0207] In one embodiment, the compositions according to the
invention additionally comprise preservatives.
[0208] In one embodiment, the preservatives are chosen from the
group consisting of m-cresol and phenol, alone or as a mixture.
[0209] In one embodiment, the concentration of the preservatives is
between 10 and 50 mM.
[0210] In one embodiment, the concentration of the preservatives is
between 10 and 40 mM.
[0211] The compositions according to the invention can additionally
comprise additives, such as tonicity agents, such as glycerol,
NaCl, mannitol and glycine.
[0212] The compositions according to the invention can additionally
comprise additives in accordance with the pharmacopoeias, such as
surfactants, for example polysorbate.
[0213] The compositions according to the invention can additionally
comprise all the excipients in accordance with the pharmacopoeias
which are compatible with the insulins used at the concentrations
of use.
[0214] In one embodiment, 0.3.ltoreq.n.ltoreq.1.7.
[0215] In one embodiment, 0.7.ltoreq.n.ltoreq.1.5.
[0216] In one embodiment, 0.9.ltoreq.n.ltoreq.1.2.
[0217] In one embodiment, 0.01.ltoreq.m.ltoreq.0.5.
[0218] In one embodiment, 0.02.ltoreq.m.ltoreq.0.4.
[0219] In one embodiment, 0.03.ltoreq.m.ltoreq.0.3.
[0220] In one embodiment, 0.05.ltoreq.m.ltoreq.0.2.
[0221] In one embodiment, 3.ltoreq.q.ltoreq.50.
[0222] In one embodiment, 3.ltoreq.q.ltoreq.40.
[0223] In one embodiment, 3.ltoreq.q.ltoreq.30.
[0224] In one embodiment, 3.ltoreq.q.ltoreq.20.
[0225] In one embodiment, 3.ltoreq.q.ltoreq.10.
[0226] In one embodiment, the -(f-[A]-COOH).sub.n radical is chosen
from the group consisting of the following sequences, f having the
meaning given above:
##STR00005##
[0227] In one embodiment, the -(g-[B]-k-[D]).sub.m radical is
chosen from the group consisting of the following sequences, g, k
and D having the meanings given above:
##STR00006##
[0228] In one embodiment, D is such that the X radical is an at
least divalent radical resulting from an amino acid.
[0229] In one embodiment, D is such that the X radical is an at
least divalent radical resulting from an amino acid chosen from the
group consisting of glycine, leucine, phenylalanine, lysine,
isoleucine, alanine, valine, aspartic acid and glutamic acid.
[0230] The radicals resulting from amino acids can be either
laevorotatory or dextrorotatory.
[0231] In one embodiment, D is such that the X radical is an at
least divalent radical resulting from a mono- or polyethylene
glycol.
[0232] In one embodiment, D is such that the X radical is an at
least divalent radical resulting from ethylene glycol.
[0233] In one embodiment, D is such that the X radical is an at
least divalent radical resulting from a polyethylene glycol chosen
from the group consisting of diethylene glycol and triethylene
glycol.
[0234] In one embodiment, D is such that the X radical is an at
least divalent radical resulting from a mono- or polyethylene
glycol amine.
[0235] In one embodiment, D is such that the X radical is an at
least divalent radical resulting from a mono- or polyethylene
glycol amine chosen from the group consisting of ethanolamine,
diethylene glycol amine and triethylene glycol amine.
[0236] In one embodiment, D is such that the X radical is an at
least divalent radical resulting from a mono- or polyethylene
glycol diamine.
[0237] In one embodiment, D is such that the X radical is an at
least divalent radical resulting from ethylenediamine.
[0238] In one embodiment, D is such that the X radical is an at
least divalent radical resulting from a mono- or polyethylene
glycol diamine chosen from the group consisting of diethylene
glycol diamine and triethylene glycol diamine.
[0239] In one embodiment, D is such that the Y group is an alkyl
group resulting from a hydrophobic alcohol.
[0240] In one embodiment, D is such that the Y group is an alkyl
group resulting from a hydrophobic alcohol chosen from the group
consisting of octanol (capryl alcohol), 3,7-dimethyloctan-1-ol,
decanol (decyl alcohol), dodecanol (lauryl alcohol), tetradecanol
(myristyl alcohol) and hexadecanol (cetyl alcohol).
[0241] In one embodiment, D is such that the Y group is an alkyl
group resulting from a hydrophobic acid.
[0242] In one embodiment, D is such that the Y group is an alkyl
group resulting from a hydrophobic acid chosen from the group
consisting of decanoic acid, dodecanoic acid, tetradecanoic acid
and hexadecanoic acid.
[0243] In one embodiment, D is such that the Y group is a group
resulting from a sterol.
[0244] In one embodiment, D is such that the Y group is a group
resulting from a sterol chosen from the group consisting of
cholesterol and its derivatives.
[0245] In one embodiment, D is such that the Y group is a group
resulting from a tocopherol.
[0246] In one embodiment, D is such that the Y group is a group
resulting from a tocopherol derivative chosen from the racemate,
the L isomer or the D isomer of a-tocopherol.
[0247] In one embodiment, D is such that the X radical results from
glycine, p=1, the Y group results from octanol and the functional
group l is an ester functional group.
[0248] In one embodiment, D is such that the X radical results from
glycine, p=1, the Y group results from dodecanol and the functional
group l is an ester functional group.
[0249] In one embodiment, D is such that the X radical results from
glycine, p=1, the Y group results from hexadecanol and the
functional group l is an ester functional group.
[0250] In one embodiment, D is such that the X radical results from
phenylalanine, p=1, the Y group results from octanol and the
functional group l is an ester functional group.
[0251] In one embodiment, D is such that the X radical results from
phenylalanine, p=1, the Y group results from 3,7-dimethyloctan-1-ol
and the functional group l is an ester functional group.
[0252] In one embodiment, D is such that the X radical results from
aspartic acid, p=2, the Y groups result from octanol and the
functional groups l are ester functional groups.
[0253] In one embodiment, D is such that the X radical results from
aspartic acid, p=2, the Y groups result from decanol and the
functional groups l are ester functional groups.
[0254] In one embodiment, D is such that the X radical results from
aspartic acid, p=2, the Y groups result from dodecanol and the
functional groups l are ester functional groups.
[0255] In one embodiment, D is such that the X radical results from
ethylenediamine, the Y group results from dodecanoic acid and the
functional group l is an amide functional group.
[0256] In one embodiment, D is such that the X radical results from
diethylene glycol amine, p=1, the Y group results from dodecanoic
acid and the functional group l is an ester functional group.
[0257] In one embodiment, D is such that the X radical results from
triethylene glycol diamine, p=1, the Y group results from
dodecanoic acid and the functional group l is an amide functional
group.
[0258] In one embodiment, D is such that the X radical results from
triethylene glycol diamine, p=1, the Y group results from
hexadecanoic acid and the functional group l is an amide functional
group.
[0259] In one embodiment, D is such that the X radical results from
leucine, p=1, the Y group results from cholesterol and the
functional group l is an ester functional group.
[0260] In one embodiment, D is such that X results from
ethylenediamine, p=1, the Y group results from cholesterol and the
functional group l is a carbamate functional group.
[0261] In one embodiment, the E radical is an at least divalent
radical resulting from an amino acid chosen from the group
consisting of glycine, leucine, phenylalanine, lysine, isoleucine,
alanine, valine, serine, threonine, aspartic acid and glutamic
acid.
[0262] The radicals resulting from amino acids can be either
laevorotatory or dextrorotatory.
[0263] In one embodiment, the E radical is an at least divalent
radical resulting from a mono- or polyethylene glycol amine.
[0264] In one embodiment, the E radical is an at least divalent
radical resulting from a mono- or polyethylene glycol amine chosen
from the group consisting of ethanolamine, diethylene glycol amine
and triethylene glycol amine.
[0265] In one embodiment, the E radical is an at least divalent
radical resulting from a mono- or polyethylene glycol diamine.
[0266] In one embodiment, the E radical is an at least divalent
radical resulting from ethylenediamine.
[0267] In one embodiment, the E radical is an at least divalent
radical resulting from a mono- or polyethylene glycol diamine
chosen from the group consisting of diethylene glycol diamine and
triethylene glycol diamine.
[0268] In one embodiment, the F group is an alkyl group resulting
from a hydrophobic alcohol.
[0269] In one embodiment, the F group is a group resulting from a
hydrophobic alcohol chosen from the group consisting of dodecanol
(lauryl alcohol), tetradecanol (myristyl alcohol) and hexadecanol
(cetyl alcohol).
[0270] In one embodiment, the F group is a group resulting from a
hydrophobic acid.
[0271] In one embodiment, the F group is a group resulting from a
hydrophobic acid chosen from the group consisting of dodecanoic
acid, tetradecanoic acid and hexadecanoic acid.
[0272] In one embodiment, the F group is a group resulting from a
sterol.
[0273] In one embodiment, the F group is a group resulting from a
sterol chosen from the group consisting of cholesterol and its
derivatives.
[0274] In one embodiment, the F group is a group resulting from a
tocopherol.
[0275] In one embodiment, the F group is a group resulting from a
tocopherol derivative chosen from the racemate, the L isomer or the
D isomer of a-tocopherol.
[0276] In one embodiment, the E radical results from
ethylenediamine, t=1, o is a carbamate functional group and the F
group results from cholesterol.
[0277] In one embodiment: [0278] -(f-[A]-COOH).sub.n is such that A
is the --CH.sub.2-- radical and f is an ether functional group;
[0279] -(g-[B]-k-[D]).sub.m is such that g is an ether functional
group, B is the --CH.sub.2-- radical, k is an amide functional
group and D is such that X results from glycine, l is an ester
functional group and Y results from octanol; [0280] q=38, n=0.9 and
m=0.2.
[0281] In one embodiment: [0282] -(f-[A]-COOH).sub.n is such that A
is the --CH.sub.2-- radical and f is an ether functional group;
[0283] -(g-[B]-k-[D]).sub.m is such that g is an ether functional
group, B is the --CH.sub.2-- radical, k is an amide functional
group and D is such that X results from glycine, p=1, l is an ester
functional group and Y results from hexadecanol; [0284] q=19, n=1.0
and m=0.1.
[0285] In one embodiment: [0286] -(f-[A]-COOH).sub.n is such that A
is the --CH.sub.2-- radical and f is an ether functional group;
[0287] -(g-[B]-k-[D]).sub.m is such that g is an ether functional
group, B is the --CH.sub.2-- radical, k is an amide functional
group and D is such that X results from phenylalanine, p=1, l is an
ester functional group and Y results from octanol; [0288] q=38,
n=1.0 and m=0.1.
[0289] In one embodiment: [0290] -(f-[A]-COOH).sub.n is such that A
is the --CH.sub.2-- radical and f is an ether functional group;
[0291] -(g-[B]-k-[D]).sub.m is such that g is an ether functional
group, B is the --CH.sub.2-- radical, k is an amide functional
group and D is such that X results from phenylalanine, p=1, l is an
ester functional group and Y results from octanol; [0292] q=19,
n=1.0 and m=0.2.
[0293] In one embodiment: [0294] -(f-[A]-COOH).sub.n is such that A
is the --CH.sub.2-- radical and f is an ether functional group;
[0295] -(g-[B]-k-[D]).sub.m is such that g is an ether functional
group, B is the --CH.sub.2-- radical, k is an amide functional
group and D is such that X results from phenylalanine, p=1, l is an
ester functional group and Y results from 3,7-dimethyloctan-1-ol;
[0296] q=38, n=1.0 and m=0.1.
[0297] In one embodiment: [0298] -(f-[A]-COOH).sub.n is such that A
is the --CH.sub.2-- radical and f is an ether functional group;
[0299] -(g-[B]-k-[D]).sub.m is such that g is an ether functional
group, B is the --CH.sub.2-- radical, k is an amide functional
group and D is such that X results from aspartic acid, p=2, l are
ester functional groups and Y result from octanol; [0300] q=38,
n=1.05 and m=0.05.
[0301] In one embodiment: [0302] -(f-[A]-COOH).sub.n is such that A
is the --CH.sub.2-- radical and f is an ether functional group;
[0303] -(g-[B]-k-[D]).sub.m is such that g is an ether functional
group, B is the --CH.sub.2-- radical, k is an amide functional
group and D is such that X results from aspartic acid, p=2, l are
ester functional groups and Y result from decanol; [0304] q=38,
n=1.05 and m=0.05.
[0305] In one embodiment: [0306] -(f-[A]-COOH).sub.n is such that A
is the --CH.sub.2-- radical and f is an ether functional group;
[0307] -(g-[B]-k-[D]).sub.m is such that g is an ether functional
group, B is the --CH.sub.2-- radical, k is an amide functional
group and D is such that X results from aspartic acid, p=2, l are
ester functional groups and Y result from dodecanol; [0308] q=19,
n=1.05 and m=0.05.
[0309] In one embodiment: [0310] -(f-[A]-COOH).sub.n is such that A
is the --CH.sub.2-- radical and f is an ether functional group;
[0311] -(g-[B]-k-[D]).sub.m is such that g is an ether functional
group, B is the --CH.sub.2-- radical, k is an amide functional
group and D is such that X results from ethylenediamine, p=1, l is
an amide functional group and Y results from dodecanoic acid;
[0312] q=38, n=1.0 and m=0.1.
[0313] In one embodiment: [0314] -(f-[A]-COOH).sub.n is such that A
is the --CH.sub.2--CH.sub.2-- radical and f is an ester functional
group; [0315] -(g-[B]-k-[D]).sub.m is such that g is an ester
functional group, B is the --CH.sub.2--CH.sub.2-- radical, k is an
amide functional group and D is such that X results from glycine,
p=1, l is an ester functional group and Y results from dodecanol;
[0316] q=38, n=1.3 and m=0.1.
[0317] In one embodiment: [0318] -(f-[A]-COOH).sub.n is such that A
is the --CH.sub.2-- radical and f is a carbamate functional group;
[0319] -(g-[B]-k-[D]).sub.m is such that g is a carbamate
functional group, B is the --CH.sub.2-- radical, k is an amide
functional group and D is such that X results from aspartic acid,
p=2, l are ester functional groups and Y result from octanol;
[0320] q=38, n=1.3 and m=0.1.
[0321] In one embodiment: [0322] -(f-[A]-COOH).sub.n is such that A
is the --CH.sub.2-- radical and f is an ether functional group;
[0323] -(g-[B]-k-[D]).sub.m is such that g is an ether functional
group, B is the --CH.sub.2-- radical, k is an amide functional
group and D is such that X results from aspartic acid, p=2, l are
ester functional groups and Y result from dodecanol; [0324] q=4,
n=0.96 and m=0.07.
[0325] In one embodiment: [0326] -(f-[A]-COOH).sub.n is such that A
is the --CH.sub.2-- radical and f is an ether functional group;
[0327] -(g-[B]-k-[D]).sub.m is such that g is an ether functional
group, B is the --CH.sub.2-- radical, k is an amide functional
group and D is such that X results from diethylene glycol amine,
p=1, l is an ester functional group and Y results from dodecanoic
acid; [0328] q=38, n=1.0 and m=0.1.
[0329] In one embodiment: [0330] -(f-[A]-COOH).sub.n is such that A
is the --CH.sub.2-- radical and f is an ether functional group;
[0331] -(g-[B]-k-[D]).sub.m is such that g is an ether functional
group, B is the --CH.sub.2-- radical, k is an amide functional
group and D is such that X results from triethylene glycol diamine,
p=1, l is an amide functional group and Y results from dodecanoic
acid; [0332] q=38, n=1.0 and m=0.1.
[0333] In one embodiment: [0334] -(f-[A]-COOH).sub.n is such that A
is the --CH.sub.2-- radical and f is an ether functional group;
[0335] -(g-[B]-k-[D]).sub.m is such that g is an ether functional
group, B is the --CH.sub.2-- radical, k is an amide functional
group and D is such that X results from triethylene glycol diamine,
p=1, l is an amide functional group and Y results from hexadecanoic
acid; [0336] q=38, n=1.05 and m=0.05.
[0337] In one embodiment: [0338] -(f-[A]-COOH).sub.n is such that A
is the --CH.sub.2-- radical and f is an ether functional group;
[0339] -(g-[B]-k-[D]).sub.m is such that g is an ether functional
group, B is the --CH.sub.2-- radical, k is an amide functional
group and D is such that X results from glycine, p=1, l is an ester
functional group and Y results from hexadecanol; [0340] q=19,
n=1.05 and m=0.05.
[0341] In one embodiment: [0342] -(f-[A]-COOH).sub.n is such that A
is the --CH.sub.2-- radical and f is an ether functional group;
[0343] -(g-[B]-k-[D]).sub.m is such that g is an ether functional
group, B is the --CH.sub.2-- radical, k is an amide functional
group and D is such that X results from glycine, p=1, l is an ester
functional group and Y results from hexadecanol; [0344] q=38,
n=0.37 and m=0.05.
[0345] In one embodiment: [0346] -(f-[A]-COOH).sub.n is such that A
is the --CH.sub.2-- radical and f is an ether functional group;
[0347] -(g-[B]-k-[D]).sub.m is such that g is an ether functional
group, B is the --CH.sub.2-- radical, k is an amide functional
group and D is such that X results from leucine, p=1, l is an ester
functional group and Y results from cholesterol; [0348] q=19,
n=1.61 and m=0.04.
[0349] In one embodiment: [0350] -(f-[A]-COOH).sub.n is such that A
is the --CH.sub.2-- radical and f is an ether functional group;
[0351] -(g-[B]-k-[D]).sub.m is such that g is an ether functional
group, B is the --CH.sub.2-- radical, k is an amide functional
group and D is such that X results from leucine, p=1, l is an ester
functional group and Y results from cholesterol; [0352] q=19,
n=1.06 and m=0.04.
[0353] In one embodiment: [0354] -(f-[A]-COOH).sub.n is such that A
is the --CH.sub.2-- radical and f is an ether functional group;
[0355] -(g-[B]-k-[D]).sub.m is such that g is an ether functional
group, B is the --CH.sub.2-- radical, k is an amide functional
group and D is such that X results from leucine, p=1, l is an ester
functional group and Y results from cholesterol; [0356] q=19,
n=0.66 and m=0.04.
[0357] In one embodiment: [0358] -(f-[A]-COOH).sub.n is such that A
is the --CH.sub.2-- radical and f is an ether functional group;
[0359] -(g-[B]-k-[D]).sub.m is such that g is an ether functional
group, B is the --CH.sub.2-- radical, k is an amide functional
group and D is such that X results from leucine, p=1, l is an ester
functional group and Y results from cholesterol; [0360] q=19,
n=0.46 and m=0.04.
[0361] In one embodiment: [0362] -(f-[A]-COOH).sub.n is such that A
is the --CH.sub.2-- radical and f is an ether functional group;
[0363] -(g-[B]-k-[D]).sub.m is such that g is an ether functional
group, B is the --CH.sub.2-- radical, k is an amide functional
group and D is such that X results from leucine, p=1, l is an ester
functional group and Y results from cholesterol; [0364] q=4, n=1.61
and m=0.05.
[0365] In one embodiment: [0366] -(f-[A]-COOH).sub.n is such that A
is the --CH.sub.2-- radical and f is an ether functional group;
[0367] -(g-[B]-k-[D]).sub.m is such that g is an ether functional
group, B is the --CH.sub.2-- radical, k is an amide functional
group and D is such that X results from ethylenediamine, p=1, l is
a carbamate functional group and Y results from cholesterol; [0368]
q=19, n=1.61 and m=0.04.
[0369] In one embodiment: [0370] -(f-[A]-COOH).sub.n is such that A
is the --CH.sub.2-- radical and f is a carbamate functional group;
[0371] -(g-[B]-k-[D]).sub.m is such that g is a carbamate
functional group, B is the --CH.sub.2-- radical, k is an amide
functional group and D is such that X results from leucine, p=1, l
is an ester functional group and Y results from cholesterol; [0372]
q=19, n=1.96 and m=0.04.
[0373] In one embodiment: [0374] -(f-[A]-COOH).sub.n is such that A
is the --CH.sub.2-- radical and f is an ether functional group;
[0375] -[E]-(o-[F]).sub.t is such that E results from
ethylenediamine, o is a carbamate functional group and F results
from cholesterol; [0376] q=19 and n=1.65.
[0377] In one embodiment: [0378] -(f-[A]-COOH).sub.n is such that A
is the --CH.sub.2-- radical and f is an ether functional group;
[0379] -(g-[B]-k-[D]).sub.m is such that g is an ether functional
group, B is the --CH.sub.2-- radical, k is an amide functional
group and D is such that X results from leucine, p=1, l is an ester
functional group and Y results from cholesterol; [0380] q=38,
n=0.99 and m=0.05.
[0381] In one embodiment: [0382] -(f-[A]-COOH).sub.n is such that A
is the --CH.sub.2-- radical and f is an ether functional group;
[0383] -(g-[B]-k-[D]).sub.m is such that g is an ether functional
group, B is the --CH.sub.2-- radical, k is an amide functional
group and D is such that X results from aspartic acid, p=2, l are
ester functional groups and Y result from dodecanol; [0384] q=4,
n=1.41 and m=0.16.
[0385] In one embodiment: [0386] -(f-[A]-COOH).sub.n is such that A
is the --CH.sub.2-- radical and f is an ether functional group;
[0387] -(g-[B]-k-[D]).sub.m is such that g is an ether functional
group, B is the --CH.sub.2-- radical, k is an amide functional
group and D is such that X results from aspartic acid, p=2, l are
ester functional groups and Y result from dodecanol; [0388] q=4,
n=1.50 and m=0.07.
[0389] In one embodiment: [0390] -(f-[A]-COOH).sub.n is such that A
is the --CH.sub.2-- radical and f is an ether functional group;
[0391] -(g-[B]-k-[D]).sub.m is such that g is an ether functional
group, B is the --CH.sub.2-- radical, k is an amide functional
group and D is such that X results from aspartic acid, p=2, l are
ester functional groups and Y result from decanol; [0392] q=4,
n=1.05 and m=0.05.
[0393] In one embodiment, the compositions according to the
invention comprise a dextran chosen from the group consisting of
the following dextrans of formula I, III or IV: [0394] Sodium
dextranmethylcarboxylate modified by octyl glycinate, [0395] Sodium
dextranmethylcarboxylate modified by cetyl glycinate, [0396] Sodium
dextranmethylcarboxylate modified by octyl phenylalaninate, [0397]
Sodium dextranmethylcarboxylate modified by 3,7-dimethyl-1-octyl
phenylalaninate, [0398] Sodium dextranmethylcarboxylate modified by
dioctyl aspartate, [0399] Sodium dextranmethylcarboxylate modified
by didecyl aspartate, [0400] Sodium dextranmethylcarboxylate
modified by dilauryl aspartate, [0401] Sodium
dextranmethylcarboxylate modified by N-(2-aminoethyl)dodecanamide,
[0402] Sodium dextransuccinate modified by lauryl glycinate, [0403]
N-(sodium methylcarboxylate) dextran carbamate modified by dioctyl
aspartate, [0404] Sodium dextranmethylcarboxylate modified by
2-(2-aminoethoxyl)ethyl dodecanoate, [0405] Sodium
dextranmethylcarboxylate modified by
2-(2-{2-[dodecanoylamino]ethoxy}ethoxy)ethylamine, [0406] Sodium
dextranmethylcarboxylate modified by
2-(2-{2-[hexadecanoylamino]ethoxy}ethoxy)ethylamine, [0407] Sodium
dextranmethylcarboxylate modified by cholesteryl leucinate, [0408]
Sodium dextranmethylcarboxylate modified by cholesteryl
1-ethylenediaminecarboxylate, [0409] N-(sodium methylcarboxylate)
dextran carbamate modified by cholesteryl leucinate.
[0410] In one embodiment, the compositions according to the
invention comprise a dextran chosen from the group consisting of
the dextran of the following formula II: [0411] Sodium
dextranmethylcarboxylate modified by cholesteryl
1-ethylenediaminecarboxylate grafted by reductive amination to the
reducing chain end.
[0412] The invention also relates to single-dose formulations at a
pH of between 6.6 and 7.8 comprising a basal insulin whose
isoelectric point is between 5.8 and 8.5 and a prandial
insulin.
[0413] The invention also relates to single-dose formulations at a
pH of between 7 and 7.8 comprising a basal insulin whose
isoelectric point is between 5.8 and 8.5 and a prandial
insulin.
[0414] In one embodiment, the basal insulin whose isoelectric point
is between 5.8 and 8.5 is insulin glargine.
[0415] In one embodiment, the prandial insulin is chosen from the
group consisting of Humulin.RTM. (human insulin) and Novolin.RTM.
(human insulin).
[0416] In one embodiment, the prandial insulin is chosen from the
group consisting of insulin lispro (Humalog.RTM.), insulin
glulisine (Apidra.RTM.) and insulin aspart (NovoLog.RTM.).
[0417] The dissolution at a pH of between 6.6 and 7.8 of the basal
insulins whose isoelectric point is between 5.8 and 8.5 by the
polysaccharides of formula I, II, III or IV can be observed and
controlled in a simple way, with the naked eye, by virtue of a
change in appearance of the solution
[0418] The dissolution at a pH of between 7 and 7.8 of the basal
insulins whose isoelectric point is between 5.8 and 8.5 by the
polysaccharides of formula I, II, III or IV can be observed and
controlled in a simple way, with the naked eye, by virtue of a
change in appearance of the solution.
[0419] Furthermore and just as importantly, the Applicant Company
has been able to confirm that a basal insulin whose isoelectric
point is between 5.8 and 8.5, dissolved in the presence of a
polysaccharide of formula I, II, III or IV, had lost none of its
slow insulin action.
[0420] The preparation of a composition according to the invention
exhibits the advantage of being able to be carried out by simple
mixing of an aqueous solution of basal insulin whose isoelectric
point is between 5.8 and 8.5, of a solution of prandial insulin and
of a polysaccharide of formula I, II, III or IV in aqueous solution
or in the lyophilized form. If necessary, the pH of the preparation
is adjusted to pH 7.
[0421] The preparation of a composition according to the invention
exhibits the advantage of being able to be carried out by simple
mixing of an aqueous solution of basal insulin whose isoelectric
point is between 5.8 and 8.5, of a polysaccharide of formula I, II,
III or IV in aqueous solution or in the lyophilized form, and of a
prandial insulin in aqueous solution or in the lyophilized
form.
[0422] In one embodiment, the mixture of basal insulin and
polysaccharide is concentrated by ultrafiltration before mixing
with the prandial insulin in aqueous solution or in the lyophilized
form.
[0423] If necessary, the composition of the mixture is adjusted in
excipients, such as glycerol, m-cresol, zinc chloride and tween, by
addition of concentrated solutions of these excipients to the
mixture. If necessary, the pH of the preparation is adjusted to
7.
DESCRIPTION OF THE FIGURES
[0424] FIGS. 1 to 6 present the results obtained in the form of
pharmacodynamic curves for glucose. The axis of the ordinates
represents the Dglucose (expressed in mM) as a function of the
post-injection time (expressed in minutes).
[0425] FIG. 1: Mean+standard deviation of the mean curves for the
sequential administrations of Apidra.RTM. and Lantus.RTM.
(.quadrature.) in comparison with a composition according to the
invention Polysaccharide 4/Lantus.RTM./Apidra.RTM. (75/25)
(.box-solid.).
[0426] FIG. 2: Apidra.RTM. Lantus.RTM. individual curves (tested on
six pigs).
[0427] FIG. 3: Polysaccharide 4/Apidra.RTM./Lantus.RTM. individual
curves (tested on six pigs).
[0428] FIG. 4: Mean+standard deviation of the mean curves for the
sequential administration of Humalog.RTM. and Lantus.RTM.
(.quadrature.) in comparison with the administration of a
composition according to the invention Polysaccharide
4/Humalog.RTM./Lantus.RTM. (.box-solid.).
[0429] FIG. 5: Humalog.RTM. Lantus.RTM. individual curves (tested
on six pigs).
[0430] FIG. 6: Polysaccharide 4/Humalog.RTM./Lantus.RTM. individual
curves (tested on five pigs).
[0431] FIGS. 7 to 12 present the results obtained in the form of
pharmacodynamic curves for glucose. The axis of the ordinates
represents the Dglucose (expressed in mM) as a function of the
post-injection time (expressed in hours).
[0432] FIG. 7: Mean+standard deviation of the mean curves for the
sequential administrations of Humalog.RTM. (100 IU/ml, 0.13 IU/kg)
and Lantus.RTM. (100 IU/ml, 0.4 IU/kg) , in comparison with a
composition according to the invention described in Example B28
(0.53 IU/kg) .
[0433] FIG. 8: Mean+standard deviation of the mean curves for the
sequential administrations of Humalog.RTM. (100 IU/ml, 0.13 IU/kg)
and Lantus.RTM. (100 IU/ml, 0.4 IU/kg) , in comparison with a
composition according to the invention described in Example B27
(0.47 IU/kg) .
[0434] FIG. 9: Mean+standard deviation of the mean curves for the
sequential administrations of Humalog (100 IU/ml, 0.13 IU/kg) and
Lantus.RTM. (100 IU/ml, 0.4 IU/kg) , in comparison with a
composition according to the invention described in Example B29
(0.53 IU/kg) .
[0435] FIG. 10: Mean+standard deviation of the mean curves for the
sequential administrations of Humalog.RTM. (100 IU/ml, 0.13 IU/kg)
and Lantus.RTM. (100 IU/ml, 0.4 IU/kg) , in comparison with a
composition according to the invention described in Example B31
(0.48 IU/kg) .
[0436] FIG. 11: Mean+standard deviation of the mean curves for the
sequential administrations of Humalog.RTM. (100 IU/ml, 0.24 IU/kg)
and Lantus.RTM. (100 IU/ml, 0.4 IU/kg) , in comparison with a
composition according to the invention described in Example B30
(0.64 IU/kg) .
[0437] FIG. 12: Mean+standard deviation of the mean curves for the
sequential administrations of Humalog.RTM. (100 IU/ml, 0.13 IU/kg)
and Lantus.RTM. (100 IU/ml, 0.4 IU/kg) , in comparison with a
composition according to the invention described in Example B32
(0.53 IU/kg) .
EXAMPLES
Part A
Polysaccharides
[0438] Table 1 below presents, in a nonlimiting way, examples of
polysaccharides which can be used in the compositions according to
the invention.
TABLE-US-00001 TABLE 1 SUBSTITUENTS POLY- -f-A-COONa- SACCHARIDES
g-B-k-D COMMON NAME Polysaccharide 1 q: 38 n: 0.9 m: 0.2
##STR00007## ##STR00008## Sodium dextran- methylcarboxylate
modified by octyl glycinate Polysaccharide 2 q: 19 n: 1.0 m: 0.1
Polysaccharide 16 q: 19 n: 1.05 m: 0.05 Polysaccharide 17 q: 38
##STR00009## ##STR00010## Sodium dextran- methylcarboxylate
modified by cetyl glycinate n: 0.37 m: 0.05 Polysaccharide 3 q: 38
n: 1.0 m: 0.1 Polysaccharide 4 q: 19 n: 1.0 m: 0.2 ##STR00011##
##STR00012## Sodium dextran- methylcarboxylate modified by octyl
phenylalaninate Polysaccharide 5 q: 38 n: 1.0 m: 0.1 ##STR00013##
##STR00014## Sodium dextran- methylcarboxylate modified by 3,7-
dimethyl-1- octyl phenylalaninate Polysaccharide 6 q: 38 n: 1.05 m:
0.05 ##STR00015## ##STR00016## Sodium dextran- methylcarboxylate
modified by dioctyl aspartate Polysaccharide 7 q: 38 n: 1.05 m:
0.05 Polysaccharide 29 q: 4 n: 1.05 m: 0.05 ##STR00017##
##STR00018## Sodium dextran- methylcarboxylate modified by didecyl
aspartate Polysaccharide 8 q: 19 n: 1.05 m: 0.05 Polysaccharide 27
q: 4 n: 1.41 m: 0.16 Polysaccharide 28 q: 4 n: 1.50 m: 0.07
##STR00019## ##STR00020## Sodium dextran- methylcarboxylate
modified by dilauryl aspartate Polysaccharide 9 q: 38 n: 1.0 m: 0.1
##STR00021## ##STR00022## Sodium dextran- methylcarboxylate
modified by N-(2- aminoethyl) dodecanamide Polysaccharide 10 q: 38
n: 1.3 m: 0.1 ##STR00023## ##STR00024## Sodium dextransuccinate
modified by lauryl glycinate Polysaccharide 11 q: 38 n: 1.3 m: 0.1
##STR00025## ##STR00026## N-(sodium methylcarboxylate) dextran
carbamate modified by dioctyl aspartate Polysaccharide 12 q: 4 n:
0.96 m: 0.07 ##STR00027## ##STR00028## Sodium dextran-
methylcarboxylate modified by dilauryl aspartate Polysaccharide 13
q: 38 n: 1.0 m: 0.1 ##STR00029## ##STR00030## Sodium dextran-
methylcarboxylate modified by 2-(2- aminoethoxy)ethyl dodecanoate
Polysaccharide 14 q: 38 n: 1.0 m: 0.1 ##STR00031## ##STR00032##
Sodium dextran- methylcarboxylate modified by 2-(2-{2-
[dodecanoylamino] ethoxy}ethoxy) ethylamine Polysaccharide 15 q: 38
n: 1.05 m: 0.05 ##STR00033## ##STR00034## Sodium dextran-
methylcarboxylate modified by 2-(2-{2- [hexadecanoylamino]
ethoxy}ethoxy) ethylamine Polysaccharide 18 q: 19 n: 1.61 m: 0.04
Polysaccharide 19 q: 19 n: 1.06 m: 0.04 Polysaccharide 20 q: 19 n:
0.66 m: 0.04 Polysaccharide 21 q: 19 n: 0.46 m: 0.04 Polysaccharide
22 q: 4 n: 1.61 m: 0.04 Polysaccharide 26 q: 38 n: 0.99
##STR00035## ##STR00036## Sodium dextran- methylcarboxylate
modified by cholesteryl leucinate m: 0.05 Polysaccharide 23 q: 19
n: 1.61 m: 0.04 ##STR00037## ##STR00038## Sodium dextran-
methylcarboxylate modified by cholesteryl 1-ethylenediamine-
carboxylate Polysaccharide 24 q: 19 n: 1.96 m: 0.04 ##STR00039##
##STR00040## N-(sodium methylcarboxylate) dextran carbamate
modified by cholesteryl leucinate Polysaccharide 25 q: 19 n: 1.65
##STR00041## ##STR00042## Sodium dextran- methylcarboxylate
modified by cholesteryl 1-ethylenediamine- carboxylate grafted by
reductive amination to the reducing chain end
Example A1
Preparation of Polysaccharide 1
[0439] 16 g (i.e., 296 mmol of hydroxyls) of dextran with a
weight-average molar mass of approximately 10 kg/mol (q=38,
Pharmacosmos) are dissolved in water at 420 g/l. 30 ml of 10N NaOH
(296 mmol) are added to the solution. The mixture is brought to
35.degree. C. and then 46 g (396 mmol) of sodium chloroacetate are
added. The temperature of the reaction mixture is brought to
60.degree. C. at 0.5.degree. C./min and then maintained at
60.degree. C. for 100 minutes. The reaction medium is diluted with
200 ml of water, neutralized with acetic acid and purified by
ultrafiltration through a 5 kDa PES membrane against 6 volumes of
water. The final solution is assayed by dry extract, to determine
the concentration of polysaccharide, and then assayed by acid/base
titration in is 50/50 (V/V) water/acetone, to determine the mean
number of methylcarboxylate units per glucoside unit.
[0440] According to the dry extract: [polysaccharide]=31.5
mg/g.
[0441] According to the acid/base titration: the mean number of
methylcarboxylate units per glucoside unit is 1.1.
[0442] The sodium dextranmethylcarboxylate solution is passed
through a Purolite resin (anionic) in order to obtain
dextranmethylcarboxylic acid, which is subsequently lyophilized for
18 hours.
[0443] Octyl glycinate, para-toluenesulphonic acid salt, is
obtained according to the process described in U.S. Pat. No.
4,826,818.
[0444] 10 g of dextranmethylcarboxylic acid (44.86 mmol of
methylcarboxylic acid) are dissolved in DMF at 60 g/l and then
cooled to 0.degree. C. 3.23 g of octyl glycinate,
para-toluenesulphonic acid salt, (8.97 mmol) are suspended in DMF
at 100 g/l. 0.91 g of triethylamine (8.97 mmol) is subsequently
added to the suspension. Once the polysaccharide solution is at
0.degree. C., a solution of NMM (5.24 g, 51.8 mmol) in DMF (530
g/l) and 5.62 g (51.8 mmol) of EtOCOCl are subsequently added.
After reacting for 10 minutes, the octyl glycinate suspension is
added. The medium is subsequently maintained at 10.degree. C. for
45 minutes. The medium is subsequently heated to 30.degree. C. A
solution of imidazole (10.38 g in 17 ml of water) and 52 ml of
water are added to the reaction medium. The polysaccharide solution
is ultrafiltered through a 10 kDa PES membrane against 15 volumes
of 0.9% NaCl solution and 5 volumes of water. The concentration of
the polysaccharide solution is determined by dry extract. A
fraction of solution is lyophilized and analyzed by .sup.1H NMR in
D.sub.2O in order to determine the degree of substitution of the
methylcarboxylates to give octyl glycinate per glucoside unit.
[0445] According to the dry extract: [Polysaccharide 1]=36.4
mg/g
[0446] According to the acid/base titration: n=0.9
[0447] According to the .sup.1H NMR: m=0.2.
Example A2
Preparation of Polysaccharide 2
[0448] Cetyl glycinate, para-toluenesulphonic acid salt, is
obtained according to the process described in U.S. Pat. No.
4,826,818.
[0449] A sodium dextranmethylcarboxylate, synthesized according to
the process described in Example A1 using a dextran with a
weight-average molecular weight of approximately 5 kg/mol (q=19,
Pharmacosmos), modified by cetyl glycinate, is obtained by a
process similar to that described in Example A1.
[0450] According to the dry extract: [Polysaccharide 2]=15.1
mg/g
[0451] According to the acid/base titration: n=1.05
[0452] According to the .sup.1H NMR: m=0.05.
Example A3
Preparation of Polysaccharide 3
[0453] Octyl phenylalaninate, para-toluenesulphonic acid salt, is
obtained according to the process described in U.S. Pat. No.
4,826,818.
[0454] A sodium dextranmethylcarboxylate, synthesized according to
the process described in Example A1 using a dextran with a
weight-average molecular weight of approximately 10 kg/mol (q=38,
Pharmacosmos), modified by octyl phenylalaninate, is obtained by a
process similar to that described in Example A1.
[0455] According to the dry extract: [Polysaccharide 3]=27.4
mg/g
[0456] According to the acid/base titration: n=1.0
[0457] According to the .sup.1H NMR: m=0.1.
Example A4
Preparation of Polysaccharide 4
[0458] A sodium dextranmethylcarboxylate, synthesized according to
the process described in Example A1 using a dextran with a
weight-average molecular weight of approximately 5 kg/mol (q=19,
Pharmacosmos), modified by octyl phenylalaninate, is obtained by a
process similar to that described in Example A3.
[0459] According to the dry extract: [Polysaccharide 4]=21.8
mg/g
[0460] According to the acid/base titration: n=1.0
[0461] According to the .sup.1H NMR: m=0.2.
Example A5
Preparation of Polysaccharide 5
[0462] 3,7-dimethyl-1-octyl phenylalaninate, para-toluenesulphonic
acid salt, is obtained according to the process described in U.S.
Pat. No. 4,826,818.
[0463] A sodium dextranmethylcarboxylate, synthesized according to
the process described in Example A1 using a dextran with a
weight-average molecular weight of approximately 10 kg/mol (q=38,
Pharmacosmos), modified by 3,7-dimethyl-1-octyl phenylalaninate, is
obtained by a process similar to that described in Example A1.
[0464] According to the dry extract: [Polysaccharide 5]=24.3
mg/g
[0465] According to the acid/base titration: n=1.0
[0466] According to the .sup.1H NMR: m=0.1.
Example A6
Preparation of Polysaccharide 6
[0467] Dioctyl aspartate, para-toluenesulphonic acid salt, is
obtained according to the process described in U.S. Pat. No.
4,826,818.
[0468] A sodium dextranmethylcarboxylate, synthesized according to
the process described in Example A1 using a dextran with a
weight-average molecular weight of approximately 10 kg/mol (q=38,
Pharmacosmos), modified by dioctyl aspartate, is obtained by a
process similar to that described in Example A1.
[0469] According to the dry extract: [Polysaccharide 6]=22.2
mg/g
[0470] According to the acid/base titration: n=1.05
[0471] According to the .sup.1H NMR: m=0.05.
Example A7
Preparation of Polysaccharide 7
[0472] Didecyl aspartate, para-toluenesulphonic acid salt, is
obtained according to the process described in U.S. Pat. No.
4,826,818.
[0473] A sodium dextranmethylcarboxylate, synthesized according to
the process described in Example A1 using a dextran with a
weight-average molecular weight of approximately 10 kg/mol (q=38,
Pharmacosmos), modified by didecyl aspartate, is obtained by a
process similar to that described in Example A1.
[0474] According to the dry extract: [Polysaccharide 7]=19.8
mg/g
[0475] According to the acid/base titration: n=1.05
[0476] According to the .sup.1H NMR: m=0.05.
Example A8
Preparation of Polysaccharide 8
[0477] Dilauryl aspartate, para-toluenesulphonic acid salt, is
obtained according to the process described in U.S. Pat. No.
4,826,818.
[0478] A sodium dextranmethylcarboxylate, synthesized according to
the process described in Example A1 using a dextran with a
weight-average molecular weight of approximately 5 kg/mol (q=19,
Pharmacosmos), modified by dilauryl aspartate is obtained by a
process similar to that described in Example A1.
[0479] According to the dry extract: [Polysaccharide 8]=22.8
mg/g
[0480] According to the acid/base titration: n=1.05
[0481] According to the .sup.1H NMR: m=0.05.
Example A9
Preparation of Polysaccharide 9
[0482] N-(2-aminoethyl)dodecanamide is obtained according to the
process described in U.S. Pat. No. 2,387,201, from the methyl ester
of dodecanoic acid (Sigma) and ethylenediamine (Roth).
[0483] A sodium dextranmethylcarboxylate, synthesized according to
the process described in Example A1 using a dextran with a
weight-average molecular weight of approximately 10 kg/mol (q=38,
Pharmacosmos), modified by N-(2-aminoethyl)dodecanamide, is
obtained by a process similar to that described in Example A1.
[0484] According to the dry extract: [Polysaccharide 9]=23.8
mg/g
[0485] According to the acid/base titration: n=1.0
[0486] According to the .sup.1H NMR: m=0.1.
Example A10
Preparation of Polysaccharide 10
[0487] Sodium dextransuccinate is obtained from a dextran with a
weight-average molar mass of approximately 10 kg/mol (q=38,
Pharmacosmos) according to the method described in the paper by
Sanchez-Chaves et al., 1998 (Manuel et al., Polymer, 1998, 39 (13),
2751-2757). According to the .sup.1H NMR in D.sub.2O/NaOD, the mean
number of succinate groups per glucoside unit is 1.4.
[0488] Lauryl glycinate, para-toluenesulphonic acid salt, is
obtained according to the process described in U.S. Pat. No.
4,826,818.
[0489] A sodium dextransuccinate modified by lauryl glycinate is
obtained by a process similar to that described in Example A1.
[0490] According to the dry extract: [Polysaccharide 10]=16.1
mg/g
[0491] According to the acid/base titration: n=1.3
[0492] According to the .sup.1H NMR: m=0.1.
Example A11
Preparation of Polysaccharide 11
[0493] Dioctyl aspartate, para-toluenesulphonic acid salt, is
obtained according to the process described in U.S. Pat. No.
4,826,818.
[0494] 12 g (i.e., 0.22 mol of hydroxyls) of dextran with a
weight-average molar mass of approximately 10 kg/mol (q=38,
Pharmacosmos) are dissolved in a DMF/DMSO mixture. The mixture is
brought to 80.degree. C. with stirring. 3.32 g (0.03 mol) of
1,4-diazabicyclo[2.2.2]octane and then 14.35 g (0.11 mol) of ethyl
isocyanatoacetate are gradually introduced. After reacting for 5 h,
the medium is diluted in water and purified by diafiltration
through a 5 kDa PES membrane against 0.1N NaOH, 0.9% NaCl and
water. The final solution is assayed by dry extract, to determine
the concentration of polysaccharide; and then assayed by acid/base
titration in 50/50 (V/V) water/acetone, to determine the mean
number de N-methylcarboxylate carbamate units per glucoside
unit.
[0495] According to the dry extract: [polysaccharide]=30.5 mg/g
[0496] According to the acid/base titration: the mean number of
N-methylcarboxylate carbamate units per glucoside unit is 1.4.
[0497] An N-(sodium methylcarboxylate) dextran carbamate modified
by dioctyl aspartate is obtained by a process similar to that
described in Example A1.
[0498] According to the dry extract: [Polysaccharide 11]=17.8
mg/g
[0499] According to the acid/base titration: n=1.3
[0500] According to the .sup.1H NMR: m=0.1.
Example A12
Preparation of Polysaccharide 12
[0501] Dilauryl aspartate, para-toluenesulphonic acid salt, is
obtained according to the process described in U.S. Pat. No.
4,826,818.
[0502] A sodium dextranmethylcarboxylate, synthesized according to
the process described in Example A1 using a dextran with a
weight-average molecular weight of approximately 1 kg/mol (q=4,
Pharmacosmos), modified by dilauryl aspartate, is obtained by a
process similar to that described in Example A1.
[0503] According to the dry extract: [Polysaccharide 12]=12.3
mg/g
[0504] According to the acid/base titration: n=0.96
[0505] According to the .sup.1H NMR: m=0.07.
Example A13
Preparation of Polysaccharide 13
[0506] 2-(2-aminoethoxyl)ethyl dodecanoate, para-toluenesulphonic
acid salt, is obtained according to the process described in U.S.
Pat. No. 4,826,818.
[0507] A sodium dextranmethylcarboxylate, synthesized according to
the process described in Example A1 using a dextran with a
weight-average molecular weight of approximately 10 kg/mol (q=38,
Pharmacosmos), modified by 2-(2-amino-ethoxy)ethyl dodecanoate is
obtained, by a process similar to that described in Example A1.
[0508] According to the dry extract: [Polysaccharide 13]=25.6
mg/g
[0509] According to the acid/base titration: n=1.0
[0510] According to the .sup.1H NMR: m=0.1.
Example A14
Preparation of Polysaccharide 14
[0511] 2-(2-{2-[dodecanoylamino]ethoxy}ethoxy)ethylamine is
obtained according to the process described in U.S. Pat. No.
2,387,201, from the methyl ester of dodecanoic acid (Sigma) and
triethylene glycol diamine (Huntsman).
[0512] A sodium dextranmethylcarboxylate, synthesized according to
the process described in Example A1 using a dextran with a
weight-average molecular weight of approximately 10 kg/mol (q=38,
Pharmacosmos), modified by
2-(2-{2-[dodecanoylamino]ethoxy}ethoxy)ethylamine, is obtained by a
process similar to that described in Example A1.
[0513] According to the dry extract: [Polysaccharide 14]=24.9
mg/g
[0514] According to the acid/base titration: n=1.0
[0515] According to the .sup.1H NMR: m=0.1.
Example A15
Preparation of Polysaccharide 15
[0516] 2-(2-{2-[hexadecanoylamino]ethoxy}ethoxy)ethylamine is
obtained, according to the process described in U.S. Pat. No.
2,387,201, from the methyl ester of palmitic acid (Sigma) and
triethylene glycol diamine (Huntsman).
[0517] A sodium dextranmethylcarboxylate, synthesized according to
the process described in Example A1 using a dextran with a
weight-average molecular weight of approximately 10 kg/mol (q=38,
Pharmacosmos), modified by
2-(2-{2-[hexadecanoylamino]ethoxy}ethoxy)ethylamine is obtained by
a process similar to that described in Example A1.
[0518] According to the dry extract: [Polysaccharide 15]=22.2
mg/g
[0519] According to the acid/base titration: n=1.05
[0520] According to the .sup.1H NMR: m=0.05.
Example A16
Preparation of Polysaccharide 16
[0521] Cetyl glycinate, para-toluenesulphonic acid salt, is
obtained according to the process described in U.S. Pat. No.
4,826,818.
[0522] A sodium dextranmethylcarboxylate, synthesized according to
the process described in Example A1 using a dextran with a
weight-average molecular weight of approximately 5 kg/mol (q=19,
Pharmacosmos), modified by cetyl glycinate is obtained by a process
similar to that described in Example A1.
[0523] According to the dry extract: [Polysaccharide 16]=23
mg/g
[0524] According to the acid/base titration: n=1.05
[0525] According to the .sup.1H NMR: m=0.05.
Example A17
Preparation of Polysaccharide 17
[0526] 10 g (i.e., 185 mmol of hydroxyls) of dextran with a
weight-average molar mass of approximately 10 kg/mol (q=38,
Pharmacosmos) are dissolved in water at 420 g/l. 19 ml of 10N NaOH
(185 mmol) are added to the solution. The mixture is brought to
35.degree. C. and then 8.6 g (74 mmol) of sodium chloroacetate are
added. The temperature of the reaction mixture is brought to
60.degree. C. at 0.5.degree. C./min and is then maintained at
60.degree. C. for 100 minutes. The reaction medium is diluted with
200 ml of water, neutralized with acetic acid and purified by
ultrafiltration through a 5 kDa PES membrane against 6 volumes of
water. The final solution is assayed by dry extract, to determine
the polysaccharide concentration, and then assayed by acid/base
titration in 50/50 (V/V) water/acetone, to determine the mean
number of methylcarboxylate units per glucoside unit.
[0527] According to the dry extract: [polysaccharide]=35.1 mg/g
[0528] According to the acid/base titration: the mean number of
methylcarboxylate units per glucoside unit is 0.42.
[0529] The sodium dextranmethylcarboxylate solution is passed
through a Purolite resin (anionic) in order to obtain
dextranmethylcarboxylic acid, which is subsequently lyophilized for
18 hours.
[0530] Cetyl glycinate, para-toluenesulphonic acid salt, is
obtained according to the process described in U.S. Pat. No.
4,826,818.
[0531] A sodium dextranmethylcarboxylate modified by cetyl
glycinate is obtained by a process similar to that described in
Example A1.
[0532] According to the dry extract: [Polysaccharide 17]=18
mg/g
[0533] According to the acid/base titration: n=0.37
[0534] According to the .sup.1H NMR: m=0.05.
Example A18
Preparation of Polysaccharide 18
[0535] 10 g of sodium dextranmethylcarboxylate, characterized by a
degree of substitution as methylcarboxylate of 1.10 per glucoside
unit, are synthesized from a dextran with a weight-average molar
mass of 5 kg/mol (q=19, Pharmacosmos), according to a process
similar to that described for Polysaccharide 1, and then
lyophilized.
[0536] 8 g (i.e., 64 mmol of hydroxyls) of sodium
dextranmethylcarboxylate, characterized by a degree of substitution
as methylcarboxylate of 1.10 per glucoside unit, are dissolved in
water at 1000 g/l. 6 ml of 10N NaOH (64 mmol) are added. The
mixture is heated to 35.degree. C. and 7.6 g of sodium
chloroacetate (65 mmol) are added. The mixture is gradually brought
to a temperature of 60.degree. C., and is maintained at this
temperature for an additional 100 minutes. The mixture is diluted
with water, neutralized with acetic acid and then purified by
ultrafiltration through a 5 kDa PES membrane against water. The
final solution is assayed by dry extract, to determine the
polysaccharide concentration, and then assayed by acid/base
titration in 50/50 (V/V) water/acetone, to determine the mean
number of methylcarboxylate units per glucoside unit.
[0537] According to the dry extract: [polysaccharide]=45.8 mg/g
[0538] According to the acid/base titration: the mean number of
methylcarboxylate units per glucoside unit is 1.65.
[0539] The sodium dextranmethylcarboxylate solution is passed
through a Purolite resin (anionic) in order to obtain
dextranmethylcarboxylic acid, which is subsequently lyophilized for
18 hours.
[0540] Cholesteryl leucinate, para-toluenesulphonic acid salt, is
obtained according to the process described in U.S. Pat. No.
4,826,818.
[0541] A sodium dextranmethylcarboxylate modified by cholesteryl
leucinate, is obtained by a process similar to that described in
Example A1.
[0542] According to the dry extract: [Polysaccharide 18]=21
mg/g
[0543] According to the acid/base titration: n=1.61
[0544] According to the .sup.1H NMR: m=0.04.
Example A19
Preparation of Polysaccharide 19
[0545] A sodium dextranmethylcarboxylate, synthesized according to
the process described in Example A1 using a dextran with a
weight-average molecular weight of approximately 5 kg/mol (q=19,
Pharmacosmos), modified by cholesteryl leucinate, is obtained by a
process similar to that described in Example A1.
[0546] According to the dry extract: [Polysaccharide 19]=19.4
mg/g
[0547] According to the acid/base titration: n=1.06
[0548] According to the .sup.1H NMR: m=0.04.
Example A20
Preparation of Polysaccharide 20
[0549] 16 g (i.e., 296 mmol of hydroxyls) of dextran with a
weight-average molar mass of approximately 5 kg/mol (q=19,
Pharmacosmos) are dissolved in water at 420 g/l. 30 ml of 10N NaOH
(296 mmol) are added to this solution. The mixture is brought to
35.degree. C. and then 26 g (222 mmol) of sodium chloroacetate are
added. The temperature of the reaction medium is gradually brought
to 60.degree. C. and then maintained at 60.degree. C. for 100
minutes. The reaction medium is diluted with water, neutralized
with acetic acid and purified by ultrafiltration through a 5 kDa
PES membrane against water. The final solution is assayed by dry
extract, to determine the polysaccharide concentration, and then
assayed by acid/base titration in 50/50 (V/V) water/acetone, to
determine the mean number of methylcarboxylate units per glucoside
unit.
[0550] According to the dry extract: [polysaccharide]=33.1 mg/g
[0551] According to the acid/base titration: the mean number of
methylcarboxylate units per glucoside unit is 0.70.
[0552] The sodium dextranmethylcarboxylate solution is passed
through a Purolite resin (anionic) in order to obtain
dextranmethylcarboxylic acid, which is subsequently lyophilized for
18 hours.
[0553] A sodium dextranmethylcarboxylate modified by cholesteryl
leucinate is obtained by a process similar to that described in
Example A1.
[0554] According to the dry extract: [Polysaccharide 20]=18.9
mg/g
[0555] According to the acid/base titration: n=0.66
[0556] According to the .sup.1H NMR: m=0.04.
Example A21
Preparation of Polysaccharide 21
[0557] 16 g (i.e., 296 mmol of hydroxyls) of dextran with a
weight-average molar mass of approximately 5 kg/mol (q=19,
Pharmacosmos) are dissolved in water at 420 g/l. 30 ml of 10N NaOH
(296 mmol) are added to this solution. The mixture is brought to
35.degree. C. and then 18 g (158 mmol) of sodium chloroacetate are
added. The temperature of the reaction medium is gradually brought
to 60.degree. C. and then maintained at 60.degree. C. for 100
minutes. The reaction medium is diluted with water, neutralized
with acetic acid and purified by ultrafiltration through a 1 kDa
PES membrane against water. The final solution is assayed by dry
extract, to determine the polysaccharide concentration, and then
assayed by acid/base titration in 50/50 (V/V) water/acetone, to
determine the mean number of methylcarboxylate units per glucoside
unit.
[0558] According to the dry extract: [polysaccharide]=52.6 mg/g
[0559] According to the acid/base titration: the mean number of
methylcarboxylate units per glucoside unit is 0.50.
[0560] The sodium dextranmethylcarboxylate solution is passed
through a Purolite resin (anionic) in order to obtain
dextranmethylcarboxylic acid, which is subsequently lyophilized for
18 hours.
[0561] A sodium dextranmethylcarboxylate modified by cholesteryl
leucinate is obtained by a process similar to that described in
Example A1.
[0562] According to the dry extract: [Polysaccharide 21]=18.9
mg/g
[0563] According to the acid/base titration: n=0.46
[0564] According to the .sup.1H NMR: m=0.04.
Example A22
Preparation of Polysaccharide 22
[0565] A sodium dextranmethylcarboxylate, synthesized according to
the process described in Example A18 using a dextran with a
weight-average molecular weight of approximately 1 kg/mol (q=4,
Pharmacosmos), modified by cholesteryl leucinate, is obtained by a
process similar to that described in Example A18.
[0566] According to the dry extract: [Polysaccharide 22]=20.2
mg/g
[0567] According to the acid/base titration: n=1.61
[0568] According to the .sup.1H NMR: m=0.04.
Example A23
Preparation of Polysaccharide 23
[0569] Cholesteryl 1-ethylenediaminecarboxylate hydrochloride is
obtained according to the process described in the patent
(Akiyoshi, K et al., WO 2010/053140).
[0570] A sodium dextranmethylcarboxylate, synthesized according to
the process described in Example A18 using a dextran with a
weight-average molecular weight of approximately 5 kg/mol (q=19,
Pharmacosmos), modified by cholesteryl
1-ethylenediaminecarboxylate, is obtained by a process similar to
that described in Example A18.
[0571] According to the dry extract: [Polysaccharide 23]=20.1
mg/g
[0572] According to the acid/base titration: n=1.61
[0573] According to the .sup.1H NMR: m=0.04.
Example A24
Preparation of Polysaccharide 24
[0574] 12 g (i.e., 0.22 mol of hydroxyls) of dextran with a
weight-average molar mass of approximately 5 kg/mol (q=19,
Pharmacosmos) are dissolved in a DMF/DMSO mixture. The mixture is
brought to 80.degree. C. with stirring. 3.32 g (0.03 mol) of
1,4-diazabicyclo[2.2.2]octane and then 26.8 g (0.21 mol) of ethyl
isocyanatoacetate are gradually introduced. After reacting for 5 h,
the medium is diluted in water and purified by diafiltration
through a 5 kDa PES membrane against 0.1N NaOH, 0.9% NaCl and
water. The final solution is assayed by dry extract, to determine
the concentration of polysaccharide; and then assayed by acid/base
titration in 50/50 (V/V) water/acetone, to determine the mean
number of N-methylcarboxylate carbamate units per glucoside
unit.
[0575] According to the dry extract: [polysaccharide]=30.1 mg/g
[0576] According to the acid/base titration: the mean number of
N-methylcarboxylate carbamate units per glucoside unit is 2.0.
[0577] An N-(sodium methylcarboxylate) dextran carbamate modified
by cholesteryl leucinate is obtained by a process similar to that
described in Example A1.
[0578] According to the dry extract: [Polysaccharide 24]=17.9
mg/g
[0579] According to the acid/base titration: n=1.96
[0580] According to the .sup.1H NMR: m=0.04.
Example A25
Preparation of Polysaccharide 25
[0581] Cholesteryl 1-ethylenediaminecarboxylate hydrochlorate is
obtained according to the process described in the patent
(Akiyoshi, K et al., WO 2010/053140).
[0582] 10 g of dextran with a weight-average molar mass of
approximately 5 kg/mol (q=19, Pharmacosmos, 3.2 mmol of chain ends)
are dissolved in DMSO at 80.degree. C. 4.8 g of cholesteryl
1-ethylenediaminecarboxylate hydrochloride (9.5 mmol), 0.96 g of
triethylamine (9.5 mmol) and 2.0 g of sodium cyanoborohydride (32
mmol) are added to the reaction medium, which is stirred at
80.degree. C. for 24 hours. After cooling, the mixture is
precipitated from dichloromethane and then from acetone, and dried
under vacuum. According to the .sup.1H NMR, a dextran modified at
the chain end by cholesteryl 1-ethylenediaminecarboxylate is
obtained. A sodium dextranmethylcarboxylate characterized by a
degree of substitution as methylcarboxylate of 1.65 per glucoside
unit and modified at the chain end by cholesteryl
1-ethylenediaminecarboxylate was synthesized by a process similar
to that described in Example A18 using the dextran modified at the
chain end by cholesteryl 1-ethylenediaminecarboxylate.
[0583] According to the dry extract: [Polysaccharide 25]=13.7
mg/g
[0584] According to the acid/base titration: n=1.65
[0585] According to the .sup.1H NMR: each polymer chain carries a
cholesteryl 1-ethylenediaminecarboxylate group grafted to the
reducing chain end.
Example A26
Preparation of Polysaccharide 26
[0586] Cholesterol leucinate, para-toluenesulphonic acid salt, is
obtained according to the process described in U.S. Pat. No.
4,826,818.
[0587] A sodium dextranmethylcarboxylate, synthesized according to
the process described in Example A1 using a dextran with a
weight-average molecular weight of approximately 10 kg/mol (q=38,
Pharmacosmos), modified by cholesterol leucinate, is obtained by a
process similar to that described in Example A1.
[0588] According to the dry extract: [Polysaccharide 26]=26.6
mg/g
[0589] According to the acid/base titration: n=0.99
[0590] According to the .sup.1H NMR: m=0.05.
Example A27
Preparation of Polysaccharide 27
[0591] Dilauryl aspartate, para-toluenesulphonic acid salt, is
obtained according to the process described in U.S. Pat. No.
4,826,818.
[0592] A sodium dextranmethylcarboxylate, synthesized according to
the process described in Example A18 using a dextran with a
weight-average molecular weight of approximately 1 kg/mol (q=4,
Pharmacosmos), modified by dilauryl aspartate, is obtained by a
process similar to that described in Example A1.
[0593] According to the dry extract: [Polysaccharide 27]=16.7
mg/g
[0594] According to the acid/base titration: n=1.41
[0595] According to the .sup.1H NMR: m=0.16.
Example A28
Preparation of Polysaccharide 28
[0596] Dilauryl aspartate, para-toluenesulphonic acid salt, is
obtained according to the process described in U.S. Pat. No.
4,826,818.
[0597] A sodium dextranmethylcarboxylate, synthesized according to
the process described in Example A18 using a dextran with a
weight-average molecular weight of approximately 1 kg/mol (q=4,
Pharmacosmos), modified by dilauryl aspartate, is obtained by a
process similar to that described in Example A1.
[0598] According to the dry extract: [Polysaccharide 28]=25
mg/g
[0599] According to the acid/base titration: n=1.50
[0600] According to the .sup.1H NMR: m=0.07.
Example A29
Preparation of Polysaccharide 29
[0601] Didecyl aspartate, para-toluenesulphonic acid salt, is
obtained according to the process described in U.S. Pat. No.
4,826,818.
[0602] A sodium dextranmethylcarboxylate, synthesized according to
the process described in Example A1 using a dextran with a
weight-average molecular weight of approximately 1 kg/mol (q=4,
Pharmacosmos), modified by didecyl aspartate, is obtained by a
process similar to that described in Example A1.
[0603] According to the dry extract: [Polysaccharide 29]=15
mg/g
[0604] According to the acid/base titration: n=1.05
[0605] According to the .sup.1H NMR: m=0.05.
EXAMPLES
Part B
Demonstration of the Properties of the Compositions According to
the Invention
Example B1
Solution of Rapid-Acting Insulin Analogue (NovoLog.RTM.) at 100
IU/ml
[0606] This solution is a commercial solution of insulin aspart,
sold by the company Novo Nordisk under the name of NovoLog.RTM. in
the USA and Novorapid.RTM. in Europe. This product is a
rapid-acting insulin analogue.
Example B2
Solution of Rapid-Acting Insulin Analogue (Humalog.RTM.) at 100
IU/ml
[0607] This solution is a commercial solution of insulin lispro,
sold by the company Eli Lilly under the name of Humalog.RTM.. This
product is a rapid-acting insulin analogue.
Example B3
Solution of Rapid-Acting Insulin Analogue (Apidra.RTM.) at 100
IU/ml
[0608] This solution is a commercial solution of insulin glulisine,
sold by the company Sanofi-Aventis under the name of Apidra.RTM..
This product is a rapid-acting insulin analogue.
Example B4
Solution of Slow-Acting Insulin Analogue (Lantus.RTM.) at 100
IU/ml
[0609] This solution is a commercial solution of insulin glargine,
sold by the company Sanofi-Aventis under the name of Lantus.RTM..
This product is a slow-acting insulin analogue.
Example B5
Solution of Human Insulin (ActRapid.RTM.) at 100 IU/ml
[0610] This solution is a commercial solution from Novo Nordisk,
sold under the name of Actrapid.RTM.. This product is a human
insulin.
Example B6
Dissolution of Lantus.RTM. at 100 IU/ml and at pH 7 Using a
Substituted Dextran
[0611] 20 mg of Polysaccharide 4 described in Example A4 are
weighed out accurately. This lyophilisate is taken up in 2 ml of
Lantus.RTM. in its commercial formulation. A transient precipitate
appears but the solution becomes clear after approximately 30
minutes. The pH of this solution is 6.3. The pH is adjusted to 7
with a 0.1N sodium hydroxide solution. This clear solution is
filtered through a 0.22 .mu.m filter and is then placed at
+4.degree. C.
Example B7
Preparation of a Substituted Dextran/Lantus.RTM./Apidra.RTM. 75/25
Composition at pH 7
[0612] 0.25 ml of Apidra.RTM. (in its commercial formulation) is
added to 0.75 ml of the Polysaccharide 4/Lantus.RTM. solution
prepared in Example B6, in order to form 1 ml of a composition at
pH 7. The composition is clear, testifying to the good solubility
of Lantus.RTM. and Apidra.RTM. under these formulation conditions.
This clear solution is filtered through a 0.22 .mu.m filter and is
then placed at +4.degree. C.
Example B8
Preparation of a Substituted Dextran/Lantus.RTM./Humalog.RTM. 75/25
Composition at pH 7
[0613] 0.25 ml of Humalog.RTM. (in its commercial formulation) is
added to 0.75 ml of the Polysaccharide 4/Lantus.RTM. solution
prepared in Example B6, in order to form 1 ml of a composition at
pH 7. The composition is clear, testifying to the good solubility
of Lantus.RTM. and Humalog.RTM. under these formulation conditions.
This clear solution is filtered through a 0.22 .mu.m filter and is
then placed at +4.degree. C.
Example B9
Preparation of a Substituted Dextran/Lantus.RTM./NovoLog.RTM. 75/25
Composition at pH 7
[0614] 0.25 ml of NovoLog.RTM. (in its commercial formulation) is
added to 0.75 ml of the Polysaccharide 4/Lantus.RTM. solution
prepared in Example B6, in order to form 1 ml of a composition at
pH 7. The composition is clear, testifying to the good solubility
of Lantus.RTM. and NovoLog.RTM. under these formulation conditions.
This clear solution is filtered through a 0.22 .mu.m filter and is
then placed at +4.degree. C.
Example B10
Preparation of a Substituted Dextran/Lantus.RTM./ActRapid.RTM.
75/25 Composition at pH 7
[0615] 0.25 ml of ActRapid.RTM. (in its commercial formulation) is
added to 0.75 ml of the Polysaccharide 4/Lantus.RTM. solution
prepared in Example B6, in order to form 1 ml of a composition at
pH 7. The composition is clear, testifying to the good solubility
of Lantus.RTM. and ActRapid.RTM. under these formulation
conditions. This clear solution is filtered through a 0.22 .mu.m
filter and is then placed at +4.degree. C.
Example B11
Preparation of a Substituted Dextran/Lantus.RTM./Apidra.RTM. 60/40
Composition at pH 7
[0616] 0.4 ml of Apidra.RTM. (in its commercial formulation) is
added to 0.6 ml of the Polysaccharide 4/Lantus.RTM. solution
prepared in Example B6, in order to form 1 ml of a composition at
pH 7. The composition is clear, testifying to the good solubility
of Lantus.RTM. and Apidra.RTM. under these formulation conditions.
This clear solution is filtered through a 0.22 .mu.m filter and is
then placed at +4.degree. C.
Example B12
Preparation of a Substituted Dextran/Lantus.RTM./Apidra.RTM. 40/60
Composition at pH 7
[0617] 0.6 ml of Apidra.RTM. (in its commercial formulation) is
added to 0.4 ml of the Polysaccharide 4/Lantus.RTM. solution
prepared in Example B6, in order to form 1 ml of a composition at
pH 7. The composition is clear, testifying to the good solubility
of Lantus.RTM. and Apidra.RTM. under these formulation conditions.
This clear solution is filtered through a 0.22 .mu.m filter and is
then placed at +4.degree. C.
Example B13
Precipitation of Lantus.RTM.
[0618] 1 ml of Lantus.RTM. is added to 2 ml of a PBS solution
containing 20 mg/ml of BSA (bovine serum albumin). The PBS/BSA
mixture simulates the composition of the subcutaneous medium. A
precipitate appears, which is in good agreement with the mechanism
of operation of Lantus.RTM. (precipitation on injection due to the
increase in the pH).
[0619] Centrifuging at 4000 rev/min is carried out in order to
separate the precipitate from the supernatent. Subsequently,
Lantus.RTM. is assayed in the supernatent. The result of this is
that 86% of Lantus.RTM. is found in a precipitated form.
Example B14
Precipitation of a Substituted Dextran/Lantus.RTM. Composition
[0620] 1 ml of Polysaccharide 4/Lantus.RTM. solution prepared in
Example B6 is added to 2 ml of a PBS solution containing 20 mg/ml
of BSA (bovine serum albumin). The PBS/BSA mixture simulates the
composition of the subcutaneous medium. A precipitate appears.
[0621] Centrifuging at 4000 rev/min is carried out in order to
separate the precipitate from the supernatent. Subsequently,
Lantus.RTM. is assayed in the supernatent. The result of this is
that 85% of Lantus.RTM. is found in a precipitated form. This
percentage of precipitation of Lantus.RTM. is identical to that
obtained for the control described in Example B13.
[0622] Dissolution and precipitation tests identical to those
described in Examples B6 and B14 were carried out with other
substituted dextrans at the same concentration of 10 mg/ml of
polysaccharide per 100 IU/ml of Lantus.RTM.. 20 mg of
polysaccharide in the lyophilisate form are weighed out accurately.
This lyophilisate is taken up in 2 ml of Lantus.RTM. in its
commercial formulation. A transient precipitate appears but the
solution becomes clear after approximately 30 minutes to a few
hours (depending on the nature of the polysaccharide). The pH of
this solution is 6.3. The pH is adjusted to 7 with a 0.1N sodium
hydroxide solution. This clear solution is filtered through a 0.22
.mu.m filter and is then placed at +4.degree. C. The results are
collated in Table 2.
TABLE-US-00002 TABLE 2 Polysaccharide Dissolution of Precipitation
of % of No. Lantus .RTM. Lantus .RTM. precipitation 2 Yes Yes 85 1
Yes Yes Not measured 4 Yes Yes 87 3 Yes Yes Not measured 5 Yes Yes
94 6 Yes Yes Not measured 7 Yes Yes Not measured 8 Yes Yes Not
measured 9 Yes Yes 94 10 Yes Yes Not measured 15 Yes Yes Not
measured 14 Yes Yes Not measured 13 Yes Yes Not measured 12 Yes Yes
Not measured 11 Yes Yes Not measured 16 Yes Yes Not measured 17 Yes
Yes Not measured 18 Yes Yes Not measured 19 Yes Yes Not measured 20
Yes Yes Not measured 21 Yes Yes Not measured 22 Yes Yes Not
measured 23 Yes Yes Not measured 24 Yes Yes Not measured 25 Yes Yes
Not measured 26 Yes Yes Not measured
Example B15
Precipitation of a Substituted Dextran/Lantus.RTM./Apidra.RTM.
75/25 Composition at pH 7
[0623] 1 ml of the substituted dextran/Lantus.RTM./Apidra.RTM.
75/25 composition (containing 7.5 mg/ml of polysaccharide, 75 IU/ml
of Lantus.RTM. and 25 IU/ml of Apidra.RTM.) prepared in Example B7
is added to 2 ml of a PBS solution containing 20 mg/ml of BSA
(bovine serum albumin). The PBS/BSA mixture simulates the
composition of the subcutaneous medium. A precipitate appears.
[0624] Centrifuging at 4000 rev/min is carried out in order to
separate the precipitate from the supernatent. Subsequently,
Lantus.RTM. is assayed in the supernatent. The percentages of
precipitation of Lantus.RTM. are similar to the control described
in Example B13.
Example B16
Precipitation of Various Compositions, the Nature of the
Substituted Dextran being Varied
[0625] Other tests under the same conditions as those of Example
B15 were carried out in the presence of other substituted
dextrans.
[0626] The results are combined in the following Table 3 and it is
observed that the dissolution and the precipitation of Lantus.RTM.
are retained.
TABLE-US-00003 TABLE 3 Dissolution Percentage of Polysaccharide
Lantus .RTM./Apidra .RTM. precipitation of No. 75/25 Lantus .RTM. 2
Yes 85 1 Yes Not measured 4 Yes 87 3 Yes Not measured 5 Yes 86 6
Yes Not measured 7 Yes Not measured 8 Yes Not measured 9 Yes 86 10
Yes 85 15 Yes 87 14 Yes 86 13 Yes 88 12 Yes 91 18 Yes Not measured
19 Yes Not measured 20 Yes Not measured 21 Yes Not measured 22 Yes
Not measured 23 Yes Not measured 24 Yes Not measured 25 Yes Not
measured 26 Yes Not measured
Example B17
Precipitation of Various Compositions, the Nature of the Prandial
Insulin being Varied
[0627] Compositions are prepared by mixing 0.75 ml of the
Polysaccharide 4/Lantus.RTM. solution prepared in Example B6 with
0.25 ml of a prandial insulin in order to form 1 ml of substituted
dextran/Lantus.RTM./prandial insulin composition (containing 7.5
mg/ml of polysaccharide, 75 IU/ml of Lantus.RTM. and 25 IU/ml of
prandial insulin).
[0628] This composition is added to 2 ml of PBS solution containing
20 mg/ml of BSA (bovine serum albumin). The PBS/BSA mixture
simulates the composition of the subcutaneous medium. A precipitate
appears.
[0629] Centrifuging at 4000 rev/min is carried out in order to
separate the precipitate from the supernatent. Subsequently,
Lantus.RTM. is assayed in the supernatent. In the presence of the
four prandial insulins tested, Lantus.RTM. precipitates to
approximately 90%. This percentage of precipitation of Lantus.RTM.
is similar to the control described in Example B13; the results are
collated in Table 4.
TABLE-US-00004 TABLE 4 Dissolution Nature of the Lantus
.RTM./prandial insulin Percentage of precipitation prandial insulin
75/25 of Lantus .RTM. Apidra .RTM. Yes 88 NovoLog .RTM. Yes 92
Humalog .RTM. Yes 89 ActRaid .RTM. Yes 90
Example B18
Preparation of a Concentrated Solution of Slow-Acting Insulin
Analogue (Glargine)
[0630] A commercial solution of insulin glargine, sold by the
company Sanofi-Aventis under the name of Lantus.RTM., is
concentrated by ultrafiltration through a 3 kDa membrane made of
regenerated cellulose (Amicon.RTM. Ultra-15, sold by the company
Millipore). On conclusion of this ultrafiltration stage, the
concentration of insulin glargine is assayed in the retentate by
reverse phase liquid chromatography (RP-HPLC). The final
concentration of insulin glargine is subsequently adjusted by the
addition of commercial 100 IU/ml glargine solution in order to
obtain the desired final concentration. This process makes it
possible to obtain concentrated solutions of glargine, denoted
C.sub.glargine, at various concentrations of greater than 100
IU/ml, such as C.sub.glargine 200, 250, 300 and 333 IU/ml. The
concentrated solutions are filtered through a 0.22 .mu.m filter and
then stored at +4.degree. C.
Example B19
Dialysis of a Commercial Solution of Rapid-Acting Insulin Analogue
(Lispro)
[0631] A commercial solution of insulin lispro, sold by the company
Lilly under the name of Humalog.RTM. is dialyzed by ultrafiltration
through a 3 kDa membrane made of regenerated cellulose (Amicon.RTM.
Ultra-15, sold by the company Millipore). The dialysis is carried
out in a 1 mM phosphate buffer at pH 7. On conclusion of this
dialysis stage, the concentration C.sub.dialyzed Humalog of lispro
in the retentate is determined by reverse phase liquid
chromatography (RP-HPLC). The dialyzed solution is stored in a
freezer at -80.degree. C.
Example B20
Lyophilization of a Solution of Rapid-Acting Insulin Analogue
(Lispro) in its Commercial Form
[0632] A volume V.sub.Humalog of a solution of rapid-acting insulin
lispro at a concentration of 100 IU/ml in its commercial form is
placed in a Lyoguard.RTM. sterilized beforehand in an autoclave.
The Lyoguard.RTM. is placed in a freezer at -80.degree. C. for
approximately 1 h before being subjected to lyophilization
overnight at a temperature of 20.degree. C. and a pressure of 0.31
mbar.
[0633] The sterile lyophilisate thus obtained is stored at ambient
temperature.
Example B21
Lyophilization of a Dialyzed Commercial Solution of Rapid-Acting
Insulin Analogue (Lispro)
[0634] A volume V.sub.dialyzed Humalog of a solution of
rapid-acting insulin lispro obtained according to Example B19 at a
concentration of C.sub.dialyzed Humalog is placed in a
Lyoguard.RTM. sterilized beforehand in an autoclave. The
Lyoguard.RTM. is placed in a freezer at -80.degree. C. for
approximately 1 h before being subjected to lyophilization
overnight at a temperature of 20.degree. C. and a pressure of 0.31
mbar.
[0635] The sterile lyophilisate thus obtained is stored at ambient
temperature.
Example B22
Preparation of a Substituted Dextran/Glargine Composition at pH 7
Using a Substituted Dextran, According to a Process Using Glargine
in the Liquid Form (in Solution) and a Polysaccharide in the Solid
Form (Lyophilized)
[0636] A weight w.sub.polys. of Polysaccharide 18 is weighed out
accurately. This lyophilisate is taken up in a volume
V.sub.glargine of a concentrated solution of glargine prepared
according to Example B18, so as to obtain a composition exhibiting
a concentration of polysaccharide
C.sub.polys.(mg/ml)=w.sub.polys./V.sub.glargine and a concentration
of glargine C.sub.glargine (IU/ml). The solution is opalescent. The
pH of this solution is approximately 6.3. The pH is adjusted to 7
by addition of concentrated NaOH and then the solution is placed
under static conditions in an oven at 37.degree. C. for
approximately 1 hour. A volume V.sub.polys./glargine of this
visually clear solution is placed at +4.degree. C.
Example B23
Preparation of a Substituted Dextran/Glargine Composition at pH 7
Using a Substituted Dextran, According to a Process Using Glargine
in the Liquid Form (in Solution) and a Polysaccharide in the Liquid
Form (in Solution)
[0637] Concentrated solutions of m-cresol, glycerol and tween 20
are added to a mother solution of polysaccharide 20 at pH 7
exhibiting a concentration C.sub.polys. mother, so as to obtain a
polysaccharide solution having the concentration C.sub.polys.
mother/excipients (mg/ml) in the presence of these excipients at
contents equivalent to those described in the Lantus.RTM.
commercial solution in a 10 ml bottle.
[0638] A volume V.sub.Lantus of a commercial solution of
slow-acting insulin glargine, sold under the name of Lantus.RTM. at
a concentration de 100 IU/ml, is added to a volume V.sub.polys.
mother/excipients of a polysaccharide solution at the concentration
C.sub.polys. mother/excipients (mg/ml) in a sterile flask. A
cloudiness appears. The pH is adjusted to pH 7 by addition of 1M
NaOH and the solution is placed under static conditions in an oven
at 37.degree. C. for approximately 1 h. This visually clear
solution is placed at +4.degree. C.
Example B24
Preparation of a Concentrated Polysaccharide/Glargine Composition
at pH=7 Using a Substituted Dextran, According to a Process for the
Concentration of a Dilute Composition
[0639] A dilute Polysaccharide 20/glargine composition at pH 7
described in Example B23 is concentrated by ultrafiltration through
a 3 kDa membrane made of regenerated cellulose (Amicon.RTM.
Ultra-15, sold by the company Millipore). On conclusion of this
ultrafiltration stage, the retentate is clear and the concentration
of insulin glargine in the composition is assayed by reverse phase
chromatography (RP-HPLC). If necessary, the concentration of
insulin glargine in the composition is subsequently adjusted to the
desired value by diluting in a solution of m-cresol/glycerol/tween
20 excipients exhibiting, for each entity, a concentration
equivalent to that described in the Lantus.RTM. commercial solution
(in a 10 ml bottle). This solution at pH 7, which is visually
clear, exhibiting a glargine concentration C.sub.glargine (IU/ml)
and a polysaccharide concentration C.sub.polys.(mg/ml), is placed
at +4.degree. C.
Example B25
Preparation of a Substituted Dextran/Glargine/Lispro Composition at
pH 7, Starting from a Rapid-Acting Insulin Lispro in its Commercial
Form
[0640] A volume V.sub.polysach./glargine of polysaccharide/glargine
solution, pH 7, exhibiting a concentration of glargine
C.sub.glargine (IU/ml) and a concentration of Polysaccharide 18
C.sub.polys. (mg/ml), prepared according to Example B22, is added
to an insulin lispro lyophilisate obtained by lyophilization of a
volume V.sub.lispro, the preparation of which is described in
Example B19, so that the ratio
V.sub.polysach./glargine/V.sub.lispro=100/C.sub.lispro, where
C.sub.lispro is the concentration of lispro (IU/ml) targeted in the
composition. The solution is clear. The zinc content of the
formulation is adjusted to the desired concentration C.sub.zinc
(.mu.M) by addition of a concentrated zinc chloride solution. The
final pH is adjusted to 7 by addition of concentrated NaOH or
HCl.
[0641] The formulation is clear, testifying to the good solubility
of glargine and lispro under these formulation conditions. This
solution is filtered through a 0.22 .mu.m filter and is placed at
+4.degree. C.
Example B26
Preparation of a Substituted Dextran/Glargine/Lispro Composition at
pH 7, Starting from a Rapid-Acting Insulin Lispro Obtained by
Dialysis of a Commercial Solution
[0642] A volume V.sub.polysach./glargine of polysaccharide/glargine
solution, pH 7, exhibiting a concentration of glargine
C.sub.glargine (IU/ml) and a concentration of Polysaccharide 20
C.sub.polys. (mg/ml), prepared according to Example B24, is added
to an insulin lispro lyophilisate obtained by lyophilization of a
volume V.sub.dialyzed Humalog, the preparation of which is
described in Example B21, so that the ratio
V.sub.polysach./glargine/V.sub.dialyzed Humalog=C.sub.dialyzed
Humalog C.sub.lispro, where C.sub.dialyzed Humalog is the
concentration of lispro (IU/ml) obtained on conclusion of the
dialysis of the commercial solution, which stage is described in
Example B19, and C.sub.lispro is the concentration of lispro
(IU/ml) targeted in the composition. The solution is clear. The
zinc content of the formulation is adjusted to the desired
concentration C.sub.zinc (.mu.M) by addition of a concentrated zinc
chloride solution. The final pH is adjusted to 7 by addition of
concentrated NaOH or HCl.
[0643] The formulation is clear, testifying to the good solubility
of glargine and lispro under these formulation conditions. This
solution is filtered through a 0.22 .mu.m filter and is placed at
+4.degree. C.
Example B27
Preparation of a Substituted Dextran/Glargine/Lispro Composition at
pH 7 Exhibiting a Concentration of Glargine of 200 IU/ml and a
Concentration of Lispro of 33 IU/ml (Proportion as Percentage of
Insulin: 85/15 as Glargine/Lispro)
[0644] A concentrated 200 IU/ml glargine solution is prepared
according to Example B18. A Polysaccharide 18 (13 mg/ml)/glargine
300 IU/ml composition, pH 7, is prepared from Polysaccharide 18
according to the method of preparation described in Example B22.
This Polysaccharide 18/glargine 200 IU/ml composition is added to
an insulin lispro lyophilisate obtained by lyophilization of the
solution of rapid-acting analogue in its commercial form, according
to the method of preparation described in Example B25. The solution
is clear. The zinc content of the formulation is adjusted to the
concentration C.sub.zinc (.mu.M)=750 .mu.M by addition of a
concentrated zinc chloride solution. The final pH is adjusted to 7
by addition of concentrated NaOH or HCl.
[0645] This composition is described in Table 5.
Example B28
Preparation of a Substituted Dextran/Glargine/Lispro Composition at
pH 7 Exhibiting a Concentration of Glargine of 200 IU/ml and a
Concentration of Lispro of 66 IU/ml (Proportion as Percentage of
Insulin: 75/25 as Glargine/Lispro)
[0646] A concentrated 200 IU/ml glargine solution is prepared
according to Example B18. A Polysaccharide 18 (13 mg/ml)/glargine
300 IU/ml composition, pH 7, is prepared from Polysaccharide 18
according to the method of preparation described in Example B22.
This Polysaccharide 18/glargine 200 IU/ml composition is added to
an insulin lispro lyophilisate obtained by lyophilization of the
solution of rapid-acting analogue in its commercial form, according
to the method of preparation described in Example B25. The solution
is clear. The zinc content of the formulation is adjusted to the
concentration C.sub.zinc (.mu.M)=1500 .mu.M by addition of a
concentrated zinc chloride solution. The final pH is adjusted to 7
by addition of concentrated NaOH or HCl.
[0647] The formulation is clear, testifying to the good solubility
of glargine and lispro under these formulation conditions. This
solution is filtered through a 0.22 .mu.m filter and is placed at
+4.degree. C.
[0648] This composition is described in Table 5.
Example B29
Preparation of a Substituted Dextran/Glargine/Lispro Composition at
pH 7 Exhibiting a Concentration of Glargine of 300 IU/ml and a
Concentration of Lispro of 100 IU/ml (Proportion as Percentage of
Insulin: 75/25 as Glargine/Lispro)
[0649] A concentrated 300 IU/ml glargine solution is prepared
according to Example B18. A Polysaccharide 18 (23 mg/ml)/glargine
300 IU/ml composition, pH 7, is prepared from Polysaccharide 18
according to the method of preparation described in Example B22.
This Polysaccharide 18/glargine 300 IU/ml composition is added to
an insulin lispro lyophilisate obtained by lyophilization of the
solution of rapid-acting analogue in its commercial form, according
to the method of preparation described in Example B25. The solution
is clear. The zinc content of the formulation is adjusted to the
concentration C.sub.zinc (.mu.M)=2000 .mu.M by addition of a
concentrated zinc chloride solution. The final pH is adjusted to 7
by addition of concentrated NaOH or HCl.
[0650] The formulation is clear, testifying to the good solubility
of glargine and lispro under these formulation conditions. This
solution is filtered through a 0.22 .mu.m filter and is placed at
+4.degree. C.
[0651] This composition is described in Table 5.
Example B30
Preparation of a Substituted Dextran/Glargine/Lispro Composition at
pH 7 Exhibiting a Concentration of Glargine of 250 IU/ml and a
Concentration of Lispro of 150 IU/ml (Proportion as Percentage of
Insulin: 63/37 as Glargine/Lispro)
[0652] A concentrated 300 IU/ml glargine solution is prepared
according to Example B18. A Polysaccharide 18 (19 mg/ml)/glargine
300 IU/ml composition, pH 7, is prepared from Polysaccharide 18
according to the method of preparation described in Example B22.
This Polysaccharide 18/glargine 250 IU/ml composition is added to
an insulin lispro lyophilisate obtained by lyophilization of the
solution of rapid-acting analogue in its commercial form, according
to the method of preparation described in Example B25. The solution
is clear. The zinc content of the formulation is adjusted to the
concentration C.sub.zinc (.mu.M)=1500 .mu.M by addition of a
concentrated zinc chloride solution. The final pH is adjusted to 7
by addition of concentrated NaOH or HCl.
[0653] The formulation is clear, testifying to the good solubility
of glargine and lispro under these formulation conditions. This
solution is filtered through a 0.22 .mu.m filter and is placed at
+4.degree. C.
[0654] This composition is described in Table 5.
Example B31
Preparation of a Substituted Dextran/Glargine/Lispro Composition at
pH 7 Exhibiting a Concentration of Glargine of 333 IU/ml and a
Concentration of Lispro of 67 IU/ml (Proportion as Percentage of
Insulin: 83/17 as Glargine/Lispro)
[0655] A concentrated 333 IU/ml glargine solution is prepared
according to Example B18. A Polysaccharide 18 (20 mg/ml)/glargine
300 IU/ml composition, pH 7, is prepared from Polysaccharide 18
according to the method of preparation described in Example B22.
This Polysaccharide 18/glargine 333 IU/ml composition is added to
an insulin lispro lyophilisate obtained by lyophilization of the
solution of rapid-acting analogue in its commercial form, according
to the method of preparation described in Example B25. The solution
is clear. The zinc content of the formulation is adjusted to the
concentration C.sub.zinc (.mu.M)=2000 .mu.M by addition of a
concentrated zinc chloride solution. The final pH is adjusted to 7
by addition of concentrated NaOH or HCl.
[0656] The formulation is clear, testifying to the good solubility
of glargine and lispro under these formulation conditions. This
solution is filtered through a 0.22 .mu.m filter and is placed at
+4.degree. C.
[0657] This composition is described in Table 5.
Example B32
Preparation of a Substituted Dextran/Glargine/Lispro Composition at
pH 7 Exhibiting a Concentration of Glargine of 300 IU/ml and a
Concentration of Lispro of 100 IU/ml (Proportion as Percentage of
Insulin: 75/25 as Glargine/Lispro)
[0658] A concentrated 300 IU/ml glargine solution is prepared
according to Example B18. A Polysaccharide 19 (23 mg/ml)/glargine
300 IU/ml composition, pH 7, is prepared from Polysaccharide 19
according to the method of preparation described in Example B22.
This Polysaccharide 19/glargine 300 IU/ml composition is added to
an insulin lispro lyophilisate obtained by lyophilization of the
solution of rapid-acting analogue under its dialyzed form,
according to the method of preparation described in Example B26.
The solution is clear. The zinc content of the formulation is
adjusted to the concentration C.sub.zinc (.mu.M)=3000 .mu.M by
addition of a concentrated zinc chloride solution. The final pH is
adjusted to 7 by addition of concentrated NaOH or HCl.
[0659] The formulation is clear, testifying to the good solubility
of glargine and lispro under these formulation conditions. This
solution is filtered through a 0.22 .mu.m filter and is placed at
+4.degree. C.
[0660] This composition is described in Table 5.
Example B33
Preparation of a Substituted Dextran/Glargine/Lispro Composition at
pH 7 Exhibiting a Concentration of Glargine of 300 IU/ml and a
Concentration of Lispro of 100 IU/ml (Proportion as Percentage of
Insulin: 75/25 as Glargine/Lispro)
[0661] A Polysaccharide 20 (23 mg/ml)/glargine 300 IU/ml
composition, pH 7, is prepared from Polysaccharide 20 according to
the method of preparation described in Example B23. This
Polysaccharide 20/glargine 300 IU/ml composition is added to an
insulin lispro lyophilisate obtained by lyophilization of the
solution of rapid-acting analogue resulting from the dialysis of a
commercial solution, according to the method of preparation
described in Example B26. The solution is clear. The zinc content
of the formulation is adjusted to the concentration C.sub.zinc
(.mu.M)=1500 .mu.M by addition of a concentrated zinc chloride
solution. The final pH is adjusted to 7 by addition of concentrated
NaOH or HCl.
[0662] The formulation is clear, testifying to the good solubility
of glargine and lispro under these formulation conditions. This
solution is filtered through a 0.22 .mu.m filter and is placed at
+4.degree. C.
[0663] This composition is described in Table 5.
TABLE-US-00005 TABLE 5 Substituted dextran/glargine/lispro
compositions at pH 7 C.sub.glargine/ Example Polysaccharide
C.sub.polysach. C.sub.glargine C.sub.lispro C.sub.lispro No. No.
(mg/ml) (IU/ml) (IU/ml) (%/%) pH B27 18 13 200 33 85/15 7 B28 18 13
200 66 75/25 7 B29 18 23 300 100 75/25 7 B30 18 19 250 150 63/37 7
B31 18 20 333 67 83/17 7 B32 19 23 300 100 75/25 7 B33 20 23 300
100 75/25 7
Example B34
Precipitation of Various Substituted Dextran/Glargine/Lispro
Compositions at pH 7 Exhibiting Different Concentrations of Insulin
Glargine and Insulin Lispro and Different Relative Proportions of
the Two Insulins
[0664] 1 ml of substituted dextran/Lantus.RTM./Humalog.RTM.
composition prepared in Examples B27 to B33 is added to 2 ml of a
PBS solution containing 20 mg/ml of BSA (bovine serum albumin). The
PBS/BSA mixture simulates the composition of the subcutaneous
medium. A precipitate appears.
[0665] Centrifuging at 4000 rev/min is carried out in order to
separate the precipitate from the supernatent. Subsequently,
Lantus.RTM. is assayed in the supernatent. The percentages of
precipitation of Lantus.RTM. are similar to the control described
in Example B13. The results are summarized in Table 6.
TABLE-US-00006 TABLE 6 Dis- solution of glargine C.sub.lispro and
of Precipitation Example Polysaccharide C.sub.polysach.
C.sub.glargine (IU/ C.sub.glargine/C.sub.lispro lispro at of % No.
No. (mg/ml) (IU/ml) ml) (%/%) pH 7 glargine Precipitation B27 18 13
200 33 85/15 YES YES 96 B28 18 13 200 66 75/25 YES YES 86 B29 18 23
300 100 75/25 YES YES 91 B30 18 19 250 150 63/37 YES YES 90 B31 18
20 333 67 83/17 YES YES 93 B32 19 23 300 100 75/25 YES YES 98 B33
20 23 300 100 75/25 YES YES Not measured
Example B35
Chemical Stability of the Compositions
[0666] The substituted dextran/Lantus.RTM./prandial insulin
compositions described in Examples B7, B27, B28 and B29 and the
corresponding controls are placed at 30.degree. C. for 4 weeks.
Regulations require 95% of native (nondegraded) insulin after 4
weeks at 30.degree. C.
[0667] After 4 weeks, the formulations studied meet the
specifications defined by the regulations. The results are combined
in Table 7.
TABLE-US-00007 TABLE 7 Percentage of native Percentage of native
glargine after prandial insulin after Compositions 4 weeks at
30.degree. C. 4 weeks at 30.degree. C. Lantus .RTM. 97 na
(commercial formulation) Apidra .RTM. na 95 (commercial
formulation) Humalog .RTM. na 98 (commercial formulation) B7 96 98
B27 97 99 B28 95 97 B29 98 100
[0668] Thus, whatever the formulation studied, a percentage of
native insulin of greater than 95% is obtained, which is in
accordance with regulatory requirements.
Example B36
Injectability of the Solutions
[0669] All the compositions prepared can be injected with the
normal systems for the injection of insulin. The solutions
described in Examples B7 to B12 and the compositions described in
Examples B27 a B33 are injected without any difficulty, both with
insulin syringes equipped with 31-gauge needles and with insulin
pens from Novo Nordisk, sold under the name of Novopen.RTM.,
equipped with 31-gauge needles.
Example B37
Protocol for Measuring the Pharmacodynamics of the Insulin
Solutions
[0670] Preclinical studies were carried out on pigs for the purpose
of evaluating two compositions according to the invention:
[0671] Lantus.RTM./Apidra.RTM. (75/25), formulated with
Polysaccharide 4 (6 mg/ml), described in Example B7, and
[0672] Lantus.RTM./Humalog.RTM. (75/25), formulated with
Polysaccharide 4 (6 mg/ml), described in Example B8.
[0673] The hypoglycaemic effects of these compositions were
compared with respect to injections carried out with simultaneous
but separate injections of Lantus.RTM. (pH 4) and then of a
prandial insulin Apidra.RTM. or Humalog.RTM..
[0674] Six domesticated pigs weighing approximately 50 kg,
catheterized beforehand in the jugular vein, are deprived of food
for 2 to 3 hours before the beginning of the experiment. In the
hour preceding the injection of insulin, three blood samples are
taken in order to determine the basal glucose level.
[0675] Injection of insulin at a dose of 0.4 IU/kg is carried out
by subcutaneous injection into the neck, under the ear of the
animal, using the Novopen.RTM. insulin pen equipped with at 31 G
needle.
[0676] Blood samples are subsequently taken after 4, 8, 12, 16, 20,
30, 40, 50, 60, 90, 120, 240, 360, 480, 600, 660 and 720 minutes.
After each sample has been taken, the catheter is rinsed with a
dilute heparin solution.
[0677] A drop of blood is taken in order to determine the glycaemia
using a glucose meter. The pharmacodynamic curves of glucose are
subsequently plotted.
[0678] The results obtained are presented in the form of
pharmacodynamic curves for glucose represented in FIGS. 1 to 6.
[0679] Lantus.RTM./Apidra.RTM. (75/25), formulated with
Polysaccharide 4 (6 mg/ml).
[0680] FIG. 1: Mean+standard deviation of the mean curves for the
sequential administrations of Apidra.RTM. and Lantus.RTM., in
comparison with a composition according to the invention
Polysaccharide 4/Lantus.RTM./Apidra.RTM. (75/25).
[0681] FIG. 2: Apidra.RTM. Lantus.RTM. individual curves.
[0682] FIG. 3: Polysaccharide 4/Apidra.RTM./Lantus.RTM. individual
curves.
[0683] FIG. 1 presents the mean curves for drop in glycaemia and
the standard deviations of the mean for the pigs tested for each
formulation. The drop in glycaemia in the first 30 minutes is
similar for the two formulations, indicating that the presence of a
polysaccharide does not interfere with the rapid-acting nature of
Apidra.RTM..
[0684] On the other hand, between 90 min and 10 h (600 minutes),
the sequential administration of Apidra.RTM. and Lantus.RTM. brings
about a heterogeneous drop in glucose with a homogeneous plateau
response with regard to three pigs and a heterogeneous response
with regard to the other three pigs (FIG. 2). In contrast, the six
pigs tested with the Polysaccharide 4/Apidra.RTM./Lantus.RTM.
formulation have a homogeneous response (FIG. 3). This is reflected
by the analysis of the coefficients of variation (CV) between 60
min and 10 h, which are on average 54% (between 21% and 113%) for
the Apidra.RTM. Lantus.RTM. control and 12% (between 5% and 25%)
for Polysaccharide 4/Apidra.RTM./Lantus.RTM..
[0685] Lantus.RTM./Humalog.RTM. (75/25), formulated with
Polysaccharide 4 (6 mg/ml).
[0686] FIG. 4: Mean+standard deviation of the mean curves for the
sequential administrations of Humalog.RTM. and Lantus.RTM. in
comparison with the administration of a composition according to
the invention Polysaccharide 4/Humalog.RTM./Lantus.RTM..
[0687] FIG. 5: Humalog.RTM. Lantus.RTM. individual curves.
[0688] FIG. 6: Polysaccharide 4/Humalog.RTM./Lantus.RTM. individual
curves.
[0689] FIG. 4 presents the mean curves for drop in glycaemia and
the standard deviations of the mean for the pigs tested with regard
to each formulation. The drop in glycaemia in the first 30 minutes
is similar for the two formulations, indicating that the presence
of Polysaccharide 4 does not interfere with the rapid-acting nature
of Humalog.RTM.. On the other hand, between 60 min and 8 h (480
minutes), the sequential administration of Humalog.RTM. and
Lantus.RTM. brings about a heterogeneous drop in glucose with a
homogeneous plateau response with regard to four pigs and a
heterogeneous response with regard to the other two pigs (FIG. 5).
In contrast, the 5 pigs tested with the Polysaccharide
4/Humalog.RTM./Lantus.RTM. formulation have a homogeneous response
(FIG. 6). This is reflected by the analysis of the coefficients of
variation (CV) with regard to the data for drop in glycaemia
between 60 min and 8 h, which are on average 54% (between 31% and
72%) for the Humalog.RTM. Lantus.RTM. control and 15% (between 6%
and 28%) for Polysaccharide 4/Humalog.RTM./Lantus.RTM.. The
presence of Polysaccharide 4 thus greatly reduced the variability
of Lantus.RTM. with regard to the drop in glycaemia.
Example B38
Protocol for Measuring the Pharmacodynamics of the Insulin
Solutions
[0690] Preclinical studies were carried out on dogs for the purpose
of evaluating six compositions according to the invention:
[0691] The hypoglycaemic effects of these compositions were
compared with respect to injections carried out with simultaneous
but separate injections of 100 IU/ml Lantus.RTM. (pH 4) and then of
a prandial insulin 100 IU/ml Humalog.RTM..
[0692] Ten domesticated dogs (beagles) weighing approximately 12 kg
are deprived of food for 18 hours before the beginning of the
experiment. In the hour preceding the injection of insulin, three
blood samples are taken in order to determine the basal glucose
level.
[0693] The injection of insulin at a dose of 0.53 IU/kg (unless
otherwise mentioned in the examples below) is carried out by
subcutaneous injection into the neck of the animal using the
Novopen.RTM. insulin pen equipped with a 31 G needle.
[0694] Blood samples are subsequently taken after 10, 20, 30, 40,
50, 60, 90, 120, 180, 240, 300, 360, 420, 480, 540, 600, 660, 720,
780, 840, 900 and 960 minutes. The first samples are taken with a
catheter (up to 180 min) and then directly from the jugular vein.
After each sample has been taken, the catheter is rinsed with a
dilute heparin solution.
[0695] A drop of blood is taken in order to determine the glycaemia
by means of a glucose meter. The pharmacodynamic curves for glucose
are subsequently plotted.
[0696] The results obtained are presented in the form of
pharmacodynamic curves for glucose represented in FIGS. 7 to
12.
The Solution of Example B28
[0697] FIG. 7: Mean+standard deviation of the mean curves for the
sequential administrations of Humalog.RTM. (100 IU/ml, 0.13 IU/kg)
and Lantus.RTM. (100 IU/ml, 0.4 IU/kg), in comparison with a
composition according to the invention described in Example B28
(0.53 IU/kg).
[0698] FIG. 7 presents the mean curves for drop in glycaemia and
the standard deviations of the mean for the dogs tested for each
formulation. The two curves are similar up to 12 hours with a rapid
drop in glycaemia, indicating that the polysaccharide does not
influence the rapid-acting effect of Humalog.RTM., a marked return
between the peak due to Humalog.RTM. and the plateau due to
glargine, and then a plateau of the glargine up to 12 h, indicating
that the basal effect of glargine is well retained.
The Solution of Example B27
[0699] FIG. 8: Mean+standard deviation of the mean curves for the
sequential administrations of Humalog.RTM. (100 IU/ml, 0.13 IU/kg)
and Lantus.RTM. (100 IU/ml, 0.4 IU/kg) in comparison with a
composition according to the invention described in Example B27
(0.47 IU/kg).
[0700] FIG. 8 presents the mean curves for drop in glycaemia and
the standard deviations of the mean for the dogs tested for each
formulation. In this comparison, the dose of basal insulin
(Lantus.RTM.) is identical whereas the dose of Humalog.RTM. is half
for the composition, with respect to the control. The drop in
glucose is greater in the case of the formulation B27 in comparison
with the control, whereas this control contains twice as much
Humalog.RTM.. On the other hand, the duration of the plateau is
shorter in the case of the combination, with respect to the
control. This indicates that, in this composition, a portion of
Lantus.RTM. is not precipitated on injection and acts with
Humalog.RTM..
The Solution of Example B29
[0701] FIG. 9: Mean+standard deviation of the mean curves for the
sequential administrations of Humalog.RTM. (100 IU/ml, 0.13 IU/kg)
and Lantus.RTM. (100 IU/ml, 0.4 IU/kg) in comparison with a
composition according to the invention described in Example B29
(0.53 IU/kg).
[0702] FIG. 9 presents the mean curves for drop in glycaemia and
the standard deviations of the mean for the dogs tested for each
formulation. The two curves are similar, with a rapid drop in
glycaemia, indicating that the polysaccharide does not influence
the rapid-acting effect of Humalog.RTM., a marked return between
the peak due to Humalog.RTM. and the plateau due to Lantus.RTM.,
and then a plateau of the Lantus.RTM. up to 13 h, indicating that
the basal effect of glargine is well retained.
The Solution of Example B31
[0703] FIG. 10: Mean+standard deviation of the mean curves for the
sequential administrations of Humalog.RTM. (100 IU/ml, 0.13 IU/kg)
and Lantus.RTM. (100 IU/ml, 0.4 IU/kg), in comparison with a
composition according to the invention described in Example B31
(0.48 IU/kg).
[0704] FIG. 10 presents the mean curves for drop in glycaemia and
the standard deviations of the mean for the dogs tested for each
formulation. In this comparison, the dose of basal insulin
(Lantus.RTM.) is identical, whereas the dose of Humalog.RTM. is
half, for the composition, with respect to the control. The drop in
glucose is greater in the case of the control, with respect to the
combination corresponding to Example B31. This response was
expected, in view of the concentration of Humalog.RTM. in the
combination, which was half that of the control. Furthermore, the
duration of the Lantus.RTM. plateau is identical in the case of the
combination, with respect to the control. This indicates that, in
this composition and by comparison with the composition described
in Example B29 (FIG. 9), it is possible to adjust the amount of
Humalog in the combination without modifying the Lantus.RTM. basal
effect.
The Solution of Example B30
[0705] FIG. 11: Mean+standard deviation of the mean curves for the
sequential administrations of Humalog.RTM. (100 IU/ml, 0.24 IU/kg)
and Lantus.RTM. (100 IU/ml, 0.4 IU/kg) in comparison with a
composition according to the invention described in Example B30
(0.64 IU/kg).
[0706] FIG. 11 presents the mean curves for drop in glycaemia and
the standard deviations of the mean for the dogs tested for each
formulation. The two curves are similar, with a rapid drop in
glycaemia, indicating that the polysaccharide does not influence
the rapid-acting effect of Humalog.RTM., a marked return between
the peak due to Humalog.RTM. and the plateau due to Lantus.RTM.,
and then a plateau of the Lantus.RTM. up to 10 h, indicating that
the glargine basal effect is well retained.
The Solution of Example B32
[0707] FIG. 12: Mean+standard deviation of the mean curves for the
sequential administrations of Humalog.RTM. (100 IU/ml, 0.13 IU/kg)
and Lantus.RTM. (100 IU/ml, 0.4 IU/kg) in comparison with a
composition according to the invention described in Example B32
(0.53 IU/kg).
[0708] FIG. 12 presents the mean curves for drop in glycaemia and
the standard deviations of the mean for the dogs tested for each
formulation. The two curves are similar up to 10 hours, with a
rapid drop in glycaemia, indicating that the polysaccharide does
not influence the rapid-acting effect of Humalog.RTM., a marked
return between the peak due to Humalog.RTM. and the plateau due to
Lantus.RTM., and then a glargine plateau, indicating that the
glargine basal effect is retained up to 10 h.
[0709] In conclusion, FIGS. 7 to 12 show that, by adjusting the
composition of the polysaccharide and the concentrations of lispro
and glargine, it is possible to obtain profiles identical to a
double injection with different proportions of rapid-acting insulin
and basal insulin. It is also possible to adjust the duration of
the basal insulin without impacting the rapid-acting insulin or to
adjust the amount of rapid-acting insulin without impacting the
effect of the basal insulin.
EXAMPLES
Part C
Demonstration of the Properties of the Compositions Comprising a
GLP-1 Analogue or Derivative According to the Invention
Example C1
0.25 mg/ml Solution of GLP-1 Analogue Exenatide (Byetta.RTM.)
[0710] This solution is an exenatide solution marketed by the
company Eli Lilly and Company under the name of Byetta.RTM..
Example C2
6 mg/ml Solution of GLP-1 Derivative Liraglutide (Victoza.RTM.)
[0711] This solution is a liraglutide solution marketed by the
company Novo Nordisk under the name of Victoza.RTM..
Example C3
Dissolution of Lantus.RTM. at 100 IU/ml and at pH 7 Using a
Substituted Dextran at a Concentration of 10 mg/ml
[0712] 20 mg of a substituted dextran chosen from those described
in is Table 1 are weighed out accurately. This lyophilisate is
taken up in 2 ml of the insulin glargine solution of Example B4 in
order to obtain a solution whose polysaccharide concentration is
equal to 10 mg/ml. After mechanically stirring on rolls at ambient
temperature, the solution becomes clear. The pH of this solution is
6.3. The pH is adjusted to 7 with a 0.1N sodium hydroxide solution.
This clear solution is filtered through a membrane (0.22 .mu.m)
filter and is then placed at +4.degree. C.
[0713] Generalization: Clear solutions of Lantus at 100 IU/ml and
at pH 7 were also obtained with concentrations of substituted
dextrans at 20 and 40 mg/ml by following the same protocol as that
described in Example C3. Thus, a weight of lyophilized
polysaccharide among those described in Table 1 is weighed out
accurately. This lyophilisate is taken up in the insulin glargine
solution of Example B4, so as to obtain a solution whose
concentration of substituted dextran is 20 or 40 mg/ml, as
described in Table 8. After mechanical stirring on rolls at ambient
temperature, the solution becomes clear. The pH of this solution is
less than 7. The pH is subsequently adjusted to 7 with a 0.1N
sodium hydroxide solution. This clear final solution is filtered
through a membrane (0.22 .mu.m) and is then placed at +4.degree.
C.
TABLE-US-00008 TABLE 8 Preparation of a solution of Lantus .RTM. at
100 IU/ml and at pH 7 using a substituted dextran at a
concentration of 10, 20 or 40 mg/ml Final concentration of Weight
of substituted Volume of the insulin substituted dextran dextran
weighed out glargine solution of (mg/ml) (mg) Example B4 added (ml)
10 20 2 20 40 2 40 80 2
Example C4
Preparation of a Lantus.RTM./Byetta.RTM. 70/30 Composition at pH
7.5
[0714] 0.09 ml of the exenatide solution of Example C1 is added to
0.21 ml of the insulin glargine solution of Example B4, in order to
obtain 0.3 ml of composition whose pH is 4.5 on mixing. The
composition, which contains 70 IU/ml of Lantus.RTM. and 0.075 mg/ml
of Byetta.RTM., is clear, testifying to the good solubility of
Lantus.RTM. and Byetta.RTM. under these formulation conditions (pH
4.5).
[0715] The pH is subsequently adjusted to 7.5 with a 0.1N sodium
hydroxide solution. The composition then becomes cloudy, testifying
to the poor solubility of the Lantus.RTM./Byetta.RTM. composition
at pH 7.5.
[0716] 70/30 Lantus.RTM./Byetta.RTM. compositions were also
prepared at pH 4.5, 5.5, 6.5, 8.5 and 9.5 by following a protocol
similar to that described in Example C4. For each of these
compositions, 0.09 ml of the exenatide solution of Example C1 is
added to 0.21 ml of the insulin glargine solution of Example B4, in
order to obtain 0.3 ml of a composition whose pH is 4.5 on mixing.
The composition is clear, testifying to the good solubility of
Lantus.RTM. and Byetta.RTM. under these formulation conditions (pH
4.5). The pH is adjusted to 5.5 or 6.5 or 8.5 or 9.5 with a 0.1N
sodium hydroxide solution. After adjusting the pH, the composition
at 5.5 is slightly cloudy, the compositions at 6.5-7.5 and 8.5 are
very cloudy and the composition at pH 9.5 is clear. These
compositions are placed at +4.degree. C. for 48 h. After 48 h at
+4.degree. C., only the composition at pH 4.5 remains clear. The
visual appearance after 48 h of the 70/30 Lantus.RTM./Byetta.RTM.
compositions at different pH values is summarized in Table 9.
TABLE-US-00009 TABLE 9 Visual appearance after 48 h of the 70/30
Lantus .RTM./Byetta .RTM. compositions at different pH values 70/30
Lantus .RTM./Byetta .RTM. compositions at different pH values pH
Visual appearance at t = 48 h 4.5 Clear 5.5 Presence of a
precipitate 6.5 Presence of a precipitate 7.5 Presence of a
precipitate 8.5 Presence of a precipitate 9.5 Presence of a
precipitate
Example C5
Preparation of the 70/30 Lantus.RTM./Victoza.RTM. Composition at pH
7.5
[0717] 0.09 ml of the liraglutide solution of Example C2 is added
to 0.21 ml of the insulin glargine solution of Example B4, in order
to obtain 0.3 ml of a composition whose pH is 7 on mixing. The
composition, which contains 70 IU/ml of glargine and 1.8 mg/ml of
liraglutide, is cloudy, testifying to the poor solubility of the
Lantus.RTM./Victoza.RTM. composition under these formulation
conditions. The pH is adjusted to 7.5 with a 0.1N sodium hydroxide
solution. After adjusting the pH, the composition remains cloudy.
This composition is placed at +4.degree. C. for 48 h.
[0718] 70/30 Lantus.RTM./Victoza.RTM. compositions were also
prepared at pH 4.5-5.5-6.5-8.5 and 9.5 by following a protocol
similar to that described in Example C5. For each of these
compositions, 0.09 ml of the liraglutide solution of Example C1 is
added to 0.21 ml of the insulin glargine solution of Example B4, in
order to obtain 0.3 ml of a composition whose pH is 7. The
composition is cloudy, testifying to the poor solubility of the
Lantus.RTM./Victoza.RTM. composition under these formulation
conditions (pH 7). The pH is adjusted to 4.5 or 5.5 or 6.5 with a
0.1N hydrochloric acid solution or to pH 9.5 with a 0.1N sodium
hydroxide solution. After adjusting the pH, the compositions at pH
4.5-5.5 and 6.5 are cloudy, testifying to the poor solubility of
the Lantus.RTM./Victoza.RTM. composition under these formulation
conditions. These compositions are placed at +4.degree. C. for 48
h. After 48 h at 4.degree. C., only the composition at pH 9.5 is
clear. The visual appearance after 48 h of the 70/30
Lantus.RTM./Victoza.RTM. compositions of different pH values is
summarized in Table 10.
TABLE-US-00010 TABLE 10 Visual appearance after 48 h of the 70/30
Lantus .RTM./Victoza .RTM. compositions at different pH values
70/30 Lantus .RTM./Victoza .RTM. compositions at different pH
values pH Visual appearance at t = 48 h 4.5 Presence of a
precipitate 5.5 Presence of a precipitate 6.5 Presence of a
precipitate 7.5 Presence of a precipitate 8.5 Presence of a
precipitate 9.5 Clear
Example C6
Preparation of a Substituted Dextran-70/30 Lantus.RTM./Byetta.RTM.
Composition at pH 7
[0719] 0.09 ml of the exenatide solution of Example C1 is added to
0.21 ml of the substituted dextran/Lantus.RTM. solution prepared in
Example C3, in order to obtain 0.3 ml of a composition at pH 5.3.
The pH is adjusted to 7 with a 0.1N sodium hydroxide solution. The
composition, which contains 7 mg/ml of polysaccharide, 70 IU/ml of
Lantus.RTM. and 0.075 mg/ml of Byetta.RTM. is clear, testifying to
the good solubility of Lantus.RTM. and Byetta.RTM. in the presence
of the substituted dextran at pH 7. This clear solution is placed
at +4.degree. C.
[0720] Generalization: Substituted dextran-Lantus.RTM./Byetta.RTM.
compositions at pH 7 were also prepared at ratios by volume
V.sub.Lantus/V.sub.Byetta of 90/10, 50/50, 30/70 and 10/90 by
following the same protocol as that described in Example C6. Thus,
a volume V.sub.Byetta of the exenatide solution of Example C1 is
added to a volume V.sub.Lantus of the substituted
dextran/Lantus.RTM. solution prepared in Example C3, in order to
obtain a composition whose pH is adjusted at 7 with a 0.1N sodium
hydroxide solution. The compositions obtained (see Table 11) are
clear, testifying to the good solubility of Lantus.RTM. and
Byetta.RTM. in the presence of a substituted dextran at pH 7. These
clear solutions are placed at +4.degree. C.
Example C7
Preparation of a Substituted Dextran-100/50 Lantus.RTM./Byetta.RTM.
Composition at pH 7
[0721] 0.150 ml of the exenatide solution of Example C1 is
lyophilized and then 0.3 ml of a substituted dextran/Lantus.RTM.
solution prepared in Example C3 are added to the lyophilisate in
order to obtain a composition whose pH is adjusted to 7 with a 0.1N
sodium hydroxide solution. The composition, which contains 10 mg/ml
of polysaccharide, 100 IU/ml of Lantus.RTM. and 0.125 mg/ml of
Byetta.RTM., is clear, testifying to the good solubility of
Lantus.RTM. and Byetta.RTM. in the presence of the substituted
dextran at pH 7. This clear solution is placed at +4.degree. C.
TABLE-US-00011 TABLE 11 Final concentrations of Lantus .RTM.,
substituted dextran and Byetta .RTM. of the compositions obtained
in Examples C6 and C7 Lantus .RTM. [Polysaccharide No.] Byetta
.RTM. IU/ml mg/ml (mg/ml) (mg/ml) 100/50 100 3.5 10 0.125 90/10 90
3.15 9 0.025 70/30 70 2.45 7 0.075 50/50 50 1.75 5 0.125 30/70 30
1.05 3 0.175
Example C8
Preparation of a Substituted Dextran-70/30 Lantus.RTM./Victoza.RTM.
Composition at pH 7
[0722] 0.09 ml of the liraglutide solution of Example C2 is added
to 0.21 ml of the substituted dextran/Lantus.RTM. solution prepared
in Example C3, in order to obtain 0.3 ml of a composition at pH
7.6. The pH is adjusted to 7 with a 0.1N hydrochloric acid
solution. The composition, which contains 7 mg/ml of
polysaccharide, 70 IU/ml of Lantus.RTM. and 1.8 mg/ml of
Victoza.RTM., is clear, testifying to the good solubility of
Lantus.RTM. and Victoza.RTM. in the presence of the substituted
dextran at pH 7. This clear solution is placed at +4.degree. C.
[0723] Generalization: Substituted dextran-Lantus.RTM./Victoza.RTM.
compositions at pH 7 have also been prepared at
V.sub.Lantus/V.sub.Victoza ratios by volume of 90/10, 50/50, 30/70,
and 90/10 by following the same protocol as that described in
Example C6. Thus, a volume V.sub.Victoza of the liraglutide
solution of Example C2 is added to a volume V.sub.Lantus of the
substituted dextran/Lantus.RTM. solution prepared in Example B3, in
order to obtain a composition whose pH is adjusted to 7 with a 0.1N
hydrochloric acid solution.
[0724] The compositions obtained (see Table 12) are clear,
testifying to the good solubility of Lantus.RTM. and Victoza.RTM.
in the presence of a substituted dextran at pH 7. These clear
solutions are placed at +4.degree. C.
Example C9
Preparation of a Substituted Dextran-100/50
Lantus.RTM./Victoza.RTM. Composition at pH 7
[0725] 0.150 ml of the liraglutide solution of Example C2 is
lyophilized and then 0.3 ml of a substituted dextran/Lantus.RTM.
solution prepared in Example C3 is added to the lyophilisate, in
order to obtain a composition whose pH is adjusted to 7 with a 0.1N
sodium hydroxide solution. The composition, which contains 10 mg/ml
of polysaccharide, 100 IU/ml of Lantus.RTM. and 3 mg/ml of
Victoza.RTM., is clear, testifying to the good solubility of
Lantus.RTM. and Victoza.RTM. in the presence of the substituted
dextran at pH 7. This clear solution is placed at +4.degree. C.
TABLE-US-00012 TABLE 12 Final concentrations of Lantus .RTM.,
substituted dextran and Victoza .RTM. of the compositions obtained
in Examples C8 and C9 Lantus .RTM. [Polysaccharide No.] Victoza
.RTM. IU/ml mg/ml (mg/ml) (mg/ml) 100/50 100 3.5 10 3 90/10 90 3.15
9 0.6 70/30 70 2.45 7 1.8 50/50 50 1.75 5 3 30/70 30 1.05 3 4.2
Example C10
Preparation of a Substituted Dextran-60/20/20
Lantus.RTM./Apidra.RTM./Byetta.RTM. Composition at pH 7
[0726] 20 mg of lyophilized Polysaccharide 4 described in Example
A3 are weighed out accurately. This lyophilisate is taken up in 2
ml of the insulin glargine solution of Example B4. After mechanical
stirring on rolls at ambient temperature, the solution becomes
clear. The pH of this solution is 6.3. The pH is adjusted to 7 with
a 0.1N sodium hydroxide solution. 0.2 ml of the exenatide solution
of Example C1 and 0.2 ml of the insulin glulisine solution of
Example B3 are added to 0.6 ml of the substituted
dextran/Lantus.RTM. solution prepared above, in order to obtain 1
ml of a composition at pH 7. The composition, which contains 6
mg/ml of polysaccharide, 60 IU/ml of Lantus.RTM., 20 IU/ml
Apidra.RTM. and 0.05 mg/ml of Byetta.RTM., is clear, testifying to
the good solubility of Lantus.RTM., Apidra.RTM. and Byetta.RTM. in
the presence of substituted dextran at pH 7. This clear solution is
filtered through a membrane (0.22 .mu.m) and is then placed at
+4.degree. C.
Example C11
Precipitation of Lantus.RTM.
[0727] 0.250 ml of Lantus.RTM. is added to 0.5 ml of a PBS
(Phosphate Buffer Solution) solution containing 20 mg/ml of BSA
(Bovine Serum Albumin). The PBS/BSA mixture simulates the
composition of the subcutaneous medium.
[0728] A precipitate appears, which is in good agreement with the
mechanism of operation of Lantus.RTM. (precipitation on injection
due to the increase in the pH).
[0729] Centrifuging at 4000 rev/min is carried out in order to
separate the precipitate from the supernatent. Subsequently,
Lantus.RTM. is assayed in the supernatent. The result of this is
that 90% of Lantus.RTM. is found in a precipitated form.
Example C12
Precipitation of a Substituted Dextran/Lantus.RTM. Composition
[0730] 0.250 ml of substituted dextran/Lantus.RTM. solution
prepared in is Example C3 is added to 0.5 ml of a PBS solution
containing 20 mg/ml of BSA. The PBS/BSA mixture simulates the
composition of the subcutaneous medium. A precipitate appears.
[0731] Centrifuging at 4000 rev/min is carried out in order to
separate the precipitate from the supernatent. Subsequently,
Lantus.RTM. is assayed in the supernatent. The result of this is
that 90% of Lantus.RTM. is found in a precipitated form. This
percentage of precipitation of Lantus.RTM. is identical to that
obtained for the control described in Example C11.
Example C13
Precipitation of a Substituted Dextran-Lantus.RTM./Byetta.RTM.
Composition
[0732] 0.250 ml of the substituted dextran-Lantus.RTM./Byetta.RTM.
composition prepared in Example C6 is added to 0.5 ml of a PBS
solution containing 20 mg/ml of BSA. The PBS/BSA mixture simulates
the composition of the subcutaneous medium. A precipitate
appears.
[0733] Centrifuging at 4000 rev/min is carried out in order to
separate the precipitate from the supernatent. Subsequently,
Lantus.RTM. and Byetta.RTM. are assayed in the supernatent. The
percentage of precipitation of Lantus.RTM. is similar to the
control described in Example C11.
Example C14
Precipitation of a Substituted Dextran-70/30
Lantus.RTM./Victoza.RTM. Composition
[0734] 0.250 ml of the substituted dextran-Lantus.RTM./Victoza.RTM.
composition prepared in Example C8 is added to 0.5 ml of a PBS
solution containing 20 mg/ml of BSA (bovine serum albumin). The
PBS/BSA mixture simulates the composition of the subcutaneous
medium. A precipitate appears.
[0735] Centrifuging at 4000 rev/min is carried out in order to
separate the precipitate from the supernatent. Subsequently,
Lantus.RTM. and Victoza.RTM. are assayed in the supernatent. The
percentage of precipitation of Lantus.RTM. is similar to the
control described in Example C11.
Example C15
Precipitation of Different Compositions, the Nature of the
Substituted Dextran being Varied
[0736] Other tests under the same conditions as those of Examples
C13 and C14 were carried out in the presence of other dextrans.
[0737] Results with at most 20 mg/ml of substituted dextran and a
70/30 Lantus.RTM./Byetta.RTM. composition are combined in the
following Table 13. It is observed that the dissolution and the
precipitation of Lantus.RTM. are retained.
TABLE-US-00013 TABLE 13 Results of the dissolution and
precipitation tests obtained with at most 20 mg/ml of substituted
dextran and a 70/30 Lantus .RTM./Byetta .RTM. composition
Dissolution Percentage of Polysaccharide 70/30 precipitation of No.
Lantus .RTM./Byetta .RTM. Lantus .RTM. 1 Yes 94 2 Yes 96 5 Yes 88 7
Yes 95 10 Yes Not measured 11 Yes 81 14 Yes Not measured 16 Yes 96
26 Yes 81 27 Yes 96 28 Yes 96 29 Yes 95
[0738] Results with at most 20 mg/ml of substituted dextran and
various Lantus.RTM./Byetta.RTM. compositions are combined in the
following Table 14. It is observed that the dissolution and the
precipitation of Lantus.RTM. are retained.
TABLE-US-00014 TABLE 14 Results of the dissolution and
precipitation tests obtained with at most 20 mg/ml of substituted
dextran and various Lantus .RTM./Byetta .RTM. compositions Ratio
Dissolution Percentage of Polysaccharide Lantus .RTM./ Lantus
.RTM./ precipitation of No. Byetta .RTM. Byetta .RTM. Lantus .RTM.
4 100/50 Yes 95 4 90/10 Yes 94 4 70/30 Yes 95 4 50/50 Yes 90 4
30/70 Yes 82 8 100/50 Yes 96 8 90/10 Yes 94 8 70/30 Yes 96 8 50/50
Yes 90 8 30/70 Yes 81
[0739] Results with at most 40 mg/ml of substituted dextran and a
70/30 Lantus.RTM./Victoza.RTM. composition are combined in the
following Table 15. It is observed that the dissolution and the
precipitation of Lantus.RTM. are retained.
TABLE-US-00015 TABLE 15 Results of the dissolution and
precipitation tests obtained with at most 40 mg/ml of substituted
dextran and a 70/30 Lantus .RTM./Victoza .RTM. composition
Dissolution Percentage of Polysaccharide 70/30 precipitation of No.
Lantus .RTM./Victoza .RTM. Lantus .RTM. 1 Yes 95 2 Yes 97 5 Yes Not
measured 7 Yes 97 10 Yes Not measured 11 Yes Not measured 14 Yes 90
16 Yes 97 26 Yes 74 27 Yes 96 28 Yes 95 29 Yes 94
[0740] Results with at most 20 mg/ml of substituted dextran and
various
[0741] Lantus.RTM./Victoza.RTM. compositions are combined in the
following Table 16. It is observed that the dissolution and the
precipitation of Lantus.RTM. are retained.
TABLE-US-00016 TABLE 16 Results of the dissolution and
precipitation tests obtained with at most 20 mg/ml of substituted
dextran and various Lantus .RTM./Victoza .RTM. compositions Ratio
Dissolution Percentage of Polysaccharide Lantus .RTM./ Lantus
.RTM./ precipitation of No. Victoza .RTM. Victoza .RTM. Lantus
.RTM. 4 90/10 Yes 94 4 70/30 Yes Not measured 4 50/50 Yes 90 4
30/70 Yes 86 8 100/50 Yes 93 8 90/10 Yes 95 8 70/30 Yes 98 8 50/50
Yes 89 8 30/70 Yes 85
Example C16
Precipitation of a Substituted Dextran-60/20/20
Lantus.RTM./Apidra.RTM./Byetta.RTM. Composition at pH 7
[0742] 0.250 ml of the substituted
dextran-Lantus.RTM./Apidra.RTM./Byetta.RTM. composition prepared in
Example C10 is added to 0.5 ml of a PBS solution containing 20
mg/ml of BSA. The PBS/BSA mixture simulates the composition of the
subcutaneous medium. A precipitate appears.
[0743] Centrifuging at 4000 rev/min is carried out in order to
separate the precipitate from the supernatent. Subsequently,
Lantus.RTM. is assayed in the supernatent. The percentage of
precipitation of Lantus.RTM. is similar to the control described in
Example C11.
* * * * *