U.S. patent application number 12/279806 was filed with the patent office on 2009-02-12 for dissolution aids for oral peptide delivery comprising a biguanide.
This patent application is currently assigned to AXCESS LIMITED. Invention is credited to Roger R.C. New.
Application Number | 20090041849 12/279806 |
Document ID | / |
Family ID | 36142051 |
Filed Date | 2009-02-12 |
United States Patent
Application |
20090041849 |
Kind Code |
A1 |
New; Roger R.C. |
February 12, 2009 |
DISSOLUTION AIDS FOR ORAL PEPTIDE DELIVERY COMPRISING A
BIGUANIDE
Abstract
A pharmaceutical composition comprising a mixture of: (c) an
active macromolecular principle; (d) an aromatic alcohol absorption
enhancer chosen from propyl gallate, butylated hydroxy toluene
(BHT), butylated hydroxy anisole (BHA) and analogues and
derivatives thereof, or mixtures thereof; and (d) a biguanide or a
pharmaceutically acceptable salt thereof, capable of increasing the
solubility of the aromatic alcohol absorption enhancer in an
aqueous medium, wherein the aromatic alcohol absorption enhancer is
present in an amount by weight greater than or equal to that of the
active principle.
Inventors: |
New; Roger R.C.; (London,
GB) |
Correspondence
Address: |
NIXON & VANDERHYE, PC
901 NORTH GLEBE ROAD, 11TH FLOOR
ARLINGTON
VA
22203
US
|
Assignee: |
AXCESS LIMITED
Jersey
GB
|
Family ID: |
36142051 |
Appl. No.: |
12/279806 |
Filed: |
February 16, 2007 |
PCT Filed: |
February 16, 2007 |
PCT NO: |
PCT/GB2007/000539 |
371 Date: |
October 17, 2008 |
Current U.S.
Class: |
424/489 ;
424/85.4; 514/1.1; 514/44R; 514/54 |
Current CPC
Class: |
A61K 31/05 20130101;
A61P 3/04 20180101; A61K 38/23 20130101; A61K 31/7088 20130101;
A61K 38/28 20130101; A61K 47/10 20130101; A61P 19/10 20180101; A61P
35/00 20180101; A61K 9/4858 20130101; A61K 2300/00 20130101; A61K
2300/00 20130101; A61K 2300/00 20130101; A61K 2300/00 20130101;
A61K 2300/00 20130101; A61K 2300/00 20130101; A61K 31/192 20130101;
A61K 2300/00 20130101; A61K 45/06 20130101; A61K 31/7088 20130101;
A61P 3/10 20180101; A61K 9/4891 20130101; A61K 38/23 20130101; A61K
31/155 20130101; A61K 31/192 20130101; A61K 31/715 20130101; A61K
31/715 20130101; A61K 38/28 20130101; A61P 3/00 20180101; A61K
31/155 20130101; A61K 31/05 20130101; A61K 47/14 20130101; A61P
19/00 20180101; A61P 43/00 20180101; A61P 19/02 20180101 |
Class at
Publication: |
424/489 ; 514/2;
514/12; 514/44; 514/54; 514/3; 514/8; 424/85.4 |
International
Class: |
A61K 9/14 20060101
A61K009/14; A61K 38/02 20060101 A61K038/02; A61K 38/16 20060101
A61K038/16; A61K 38/28 20060101 A61K038/28; A61K 38/21 20060101
A61K038/21; A61K 38/22 20060101 A61K038/22; A61P 43/00 20060101
A61P043/00; A61K 38/18 20060101 A61K038/18; A61K 38/14 20060101
A61K038/14; A61K 31/7052 20060101 A61K031/7052; A61K 31/715
20060101 A61K031/715 |
Foreign Application Data
Date |
Code |
Application Number |
Feb 17, 2006 |
GB |
0603252.8 |
Claims
1-28. (canceled)
29. A pharmaceutical composition comprising a mixture of: (a) an
active macromolecular principle; (b) an aromatic alcohol absorption
enhancer chosen from propyl gallate, butylated hydroxy toluene
(BHT), butylated hydroxy anisole (BHA) and analogues and
derivatives thereof, or mixtures thereof; and (c) a biguanide or a
pharmaceutically acceptable salt thereof, capable of increasing the
solubility of the aromatic alcohol absorption enhancer in an
aqueous medium, wherein the aromatic alcohol absorption enhancer is
present in an amount by weight greater than or equal to that of the
active principle.
30. A composition according to claim 29, which comprises less than
5% by weight of water.
31. A composition according to claim 29, wherein the composition is
coated with an enteric coating which becomes permeable at a pH from
3 to 7.
32. A composition according claim 29, wherein the mixture comprises
from 5 to 30% by weight of the aromatic alcohol absorption
enhancer.
33. A composition according to claim 29, wherein the biguanide is
chosen from metformin, phenformin and chlorhexidine and
pharmaceutically acceptable salts thereof.
34. A composition according to claim 29, wherein the mixture is in
the form of a solution or a microparticulate dispersion.
35. A composition according to claim 29, wherein the mixture is in
solid form.
36. A composition according to claim 29, wherein the active
macromolecular principle is a polypeptide or protein,
polynucleotide, polysaccharide or a mixture thereof.
37. A composition according to claim 36, where the active
macromolecular principle is chosen from calcitonin, insulin, low
molecular weight heparin, erythropoeitin, granulocyte colony
stimulating factor, interferon, C-peptide, GLP-1, human growth
hormone and parathyroid hormone or analogues or fragments
thereof.
38. A composition according to claim 36, where the active
macromolecular principle is insulin, calcitonin or parathyroid
hormone or an analogue or a derivative thereof.
39. A composition according to claim 38, wherein the active
macromolecular principle is insulin or an analogue or derivative
thereof.
40. A composition according to claim 29, for use in the therapeutic
or diagnostic treatment of the human or animal body.
41. A composition according to claim 29, wherein the analogues and
derivatives of propyl gallate are linear or branched chain
C.sub.1-12 alkyl, C.sub.1-12 alkyloxy, C.sub.1-12 alkylthio or
C.sub.2-12 alkenyl esters of gallic acid, which are optionally
substituted with halogen, linear or branched chain C.sub.1-12
alkyl, C.sub.1-12 alkyloxy, C.sub.1-12 alkylthio or C.sub.2-12
alkenyl and the analogues and derivatives of BHT or BHA are hydroxy
toluene or hydroxy anisole wherein the methyl group or the methoxy
group linked to the aromatic ring and/or the hydrogen ortho to the
hydroxy group are replaced by linear or branched chain C.sub.1-12
alkyl, C.sub.1-12 alkyloxy, C.sub.1-12 alkylthio or C.sub.2-12
alkenyl, either unsubstituted or substituted in any position by
halogen.
42. A composition according to claim 29, wherein the aromatic
alcohol absorption enhancer is chosen from propyl gallate,
butylated hydroxy toluene and butylated hydroxy anisole.
43. A method of enhancing the absorption of an active
macromolecular principle in a patient, which method comprises
administering to said patient an aromatic alcohol chosen from
propyl gallate, BHT, BHA and analogues and derivatives thereof,
together with a biguanide or a pharmaceutically acceptable salt
thereof, capable of enhancing the solubility of the aromatic
alcohol in an aqueous medium.
44. A method according to claim 43, wherein the active
macromolecular principle to be absorbed is a polypeptide or
protein, polynucleotide, polysaccharide or a mixture thereof.
45. A method according to claim 44, wherein the active
macromolecular principle to be absorbed is chosen from calcitonin,
insulin, low molecular weight heparin, erythropoeitin, granulocyte
colony stimulating factor, interferon, C-peptide, GLP-1, human
growth hormone and parathyroid hormone or analogues or fragments
thereof.
46. A method according to claim 45, wherein the active
macromolecular principle to be absorbed is insulin, calcitonin or
parathyroid hormone or analogues or fragments thereof.
47. A method according to claim 46, wherein the active
macromolecular principle to be absorbed is insulin or an analogue
or fragment thereof.
48. A method of enhancing the absorption of an active
macromolecular principle in a patient, which method comprises
administering to said patient a composition as defined in claim
29.
49. A method according to claim 48, wherein the composition
comprises less than 5% by weight of water.
50. A method according to claim 48, wherein the composition is in
the form of a solution, as a microparticulate dispersion or as a
solid.
51. A method according to claim 48, wherein the active
macromolecular principle is chosen from insulin, C-peptide, GLP-1
or a mixture thereof, and the method is for treating diabetes.
52. A method according to claim 48, wherein the active
macromolecular principle is chosen from calcitonin and PTH, and the
method is for treating osteoporosis.
53. A method according to claim 48, wherein the active
macromolecular principle is calcitonin, and the method is for
treating osteoarthritis.
54. A method according to claim 48, wherein the active
macromolecular principle is chosen from peptide YY, oxyntomodulin
and a mixture thereof, and the method is for treating obesity.
55. A method according to claim 48, wherein the active
macromolecular principle is chosen from erythropoetin, GCST, GMCSF
and mixtures thereof, and the method is for treating cancer.
Description
[0001] This invention relates to the use of certain aromatic
alcohols as absorption enhancers to facilitate the passage of
peptides, proteins and other macromolecules across the intestinal
wall, and in particular, the use of new agents to aid in
dissolution of said aromatic alcohols, in order to improve the
availability of such agents in biological fluids, where under
normal circumstances they are extremely poorly soluble.
[0002] It has previously been reported in WO 2004/091584 that
aromatic alcohols such as propyl gallate, butylated hydroxy anisole
and butylated hydroxy toluene can act as absorption enhancers for
peptides and proteins across mucosal surfaces such as the
intestine, and the action of these agents is maximized when
formulated in combination with other agents which enhance their
dissolution in aqueous media. Examples of such dissolution aids
previously cited are bile salts such as sodium deoxycholate, and
sodium chenodeoxycholate.
[0003] On the basis of the teaching from this prior art, a person
skilled in the art would conclude that, in order to achieve
satisfactory results, a dissolution aid needs to display surfactant
activity, preferably forming micelles, as do bile salts, and
indeed, one of the biological functions of bile salts in vivo is to
aid the dissolution of dietary components, particularly lipids, in
the intestine.
[0004] It has now been found, surprisingly, that a new and
unrelated class of small molecules is also able to act as
dissolution aids for poorly-soluble aromatic alcohols, in spite of
the fact that these molecules display no surfactant properties, and
have little tendency to form micelles. This class of small
molecules is comprised of substituted biguanides, of which
metformin and phenformin are typical examples.
[0005] The action of biguanides as dissolution aids appears to be
specific to aromatic alcohols, and does not extend to other classes
of molecules such as cholesterol or fatty acids, for which bile
salts are well known as solubilising agents. Consequently, one can
conclude that there is nothing about bile salts, from either a
structural or a functional point of view, which would lead a
skilled person to suppose that biguanides might also share their
properties as dissolution aids.
[0006] Biguanides may be formulated together with poorly-soluble
aromatic alcohols as excipients to yield mucosally-administered
pharmaceutical formulations containing one or more active molecular
principles, whose passage across the mucosal barrier is enhanced as
a result of being administered in combination with the
biguanide/aromatic alcohol mixture.
[0007] The invention provides a pharmaceutical composition
comprising a mixture of:
[0008] (a) an active macromolecular principle; and
[0009] (b) an aromatic alcohol absorption enhancer chosen from
propyl gallate, butylated hydroxy toluene, butylated hydroxy
anisole and analogues and derivatives thereof, and
[0010] (c) a biguanide capable of increasing the solubility of the
aromatic alcohol absorption anhancer in aqueous media,
wherein the aromatic alcohol absorption enhancer is present in an
amount by weight greater than or equal to that of the active
macromolecular principle.
[0011] The invention also provides the use, in a pharmaceutical
composition, of an aromatic alcohol chosen from propyl gallate,
butylated hydroxy toluene, butylated hydroxy anisole and analogues
and derivatives thereof together with a biguanide or a
pharmaceutically acceptable salt thereof, capable of increasing the
solubility of the aromatic alcohol in an aqueous medium as an
enhancer for the absorption of macromolecules into the body.
[0012] In a further embodiment, the invention provides the use of
an aromatic alcohol chosen from propyl gallate, butylated hydroxy
toluene, butylated hydroxy anisole and analogues and derivatives
thereof together with a biguanide or a pharmaceutically acceptable
salt thereof, capable of increasing the solubility of the aromatic
alcohol in an aqueous medium in the manufacture of a medicament
containing an active macromolecular principle, in order to enhance
absorption of the active macromolecular principle into the human or
animal body.
[0013] The common feature of the series of molecules acting as
dissolution aids described in this invention is the biguanide
nucleus, and molecules with a variety of substitutions in the
biguanide nucleus display the desired activity. In order to assess
the suitability of a biguanide for use as a dissolution aid, the
following procedure may be followed. Typically, a solution of the
biguanide is prepared in water at a concentration of 100 mg/ml,
with heating if necessary, and appropriate adjustment of pH, if a
solution is not obtained immediately. To 1 ml of the solution 25 mg
of propyl gallate is added, and the mixture warmed with shaking for
up to half an hour. If a clear solution is obtained, the
substituted biguanide may be considered suitable for use as a
dissolution aid.
[0014] The biguanides for use in the present invention will
suitably have the following formula
##STR00001##
[0015] wherein R.sup.1, R.sup.2, R.sup.3 and R.sup.4 are each
independently chosen from hydrogen, optionally substituted alkyl,
optionally substituted phenyl, ethylene glycol, diethylene glycol,
triethylene glycol and tetraethylene glycol, or one of R.sup.1,
R.sup.2, R.sup.3 and R.sup.4 may be
##STR00002##
[0016] where R.sup.5, R.sup.6 and R.sup.7 are each independently
chosen from hydrogen, optionally substituted alkyl, optionally
substituted phenyl, ethylene glycol, diethylene glycol, triethylene
glycol and tetraethylene glycol.
[0017] Substituents for the alkyl and phenyl groups include halo,
e.g. chloro, bromo, fluoro or iodo, hydroxy and amino. The alkyl
groups preferably have from 1 to 6 carbons, and may be saturated or
unsaturated, straight chain or branched. The biguanide may be
included in the composition of the invention as a pharmaceutically
acceptable salt.
[0018] Preferred biguanides for use in the present invention
include metformin, phenformin and chlorhexidine or pharmaceutically
acceptable salts thereof. The pharmaceutically acceptable salts are
suitably the chloride, bromide, iodide or salts of organic acids
such as the acetate, propionate, mesylate (methyl sulphonate) or
glucuronate.
[0019] The biguanide may be present in the composition in an amount
of at least 50% by weight, preferably from 60 to 95% and more
preferably from 80 to 90%.
[0020] The aromatic alcohol absorption enhancer may be propyl
gallate or an analogue or a derivative thereof, and, preferably is
propyl gallate. Suitable analogues and derivatives of propyl
gallate include esters of gallic acid. The esters may be linear or
branched chain C.sub.1-12 alkyl, C.sub.1-12 alkyloxy, C.sub.1-12
alkylthio or C.sub.2-12 alkenyl esters. The compounds are
optionally substituted with halogen, linear or branched chain
C.sub.1-12 alkyl, C.sub.1-12 alkyloxy, C.sub.1-12 alkylthio or
C.sub.2-12 alkenyl esters. The aromatic alcohol absorption enhancer
may also be chosen from BHT, BHA and analogues and derivatives
thereof Suitable analogues and derivatives of BHT or BHA include
analogues and derivatives of hydroxy toluene or hydroxy anisole
where the methyl group or the methoxy group linked to the aromatic
ring and/or the hydrogen ortho to the hydroxyl group are replaced
by linear or branched chain C.sub.1-12 alkyl, C.sub.1-12 alkyloxy,
C.sub.1-12 alkylthio or C.sub.2-12 alkenyl, either unsubstituted or
substituted in any position, especially by halogen atoms.
Preferably, the aromatic alcohol absorption enhancer is chosen from
propyl gallate, BHT and BHA.
[0021] The aromatic alcohols disclosed above which are used in
pharmaceutical practice as antioxidants are included at
concentrations up to 0.1% w/v of the total formulation (see entries
for individual compounds in the Handbook of Pharmaceutical
Excipients, Eds Wade & Weller, The Pharmaceutical Press, London
UK, 2.sup.nd edition 1994). It is generally considered that higher
concentrations of the compounds give no added antioxidant benefit,
and it is thus standard pharmaceutical practice to restrict the
concentration of the antioxidants in formulations to no greater
than 0.1%. When used as absorption enhancers according to the
present invention, however, the efficacy of these compounds is
concentration dependent up to a much higher level, and their
proportions in a pharmaceutical formulation are much higher than
previously described in the prior art.
[0022] For example, WO-A-0222158 provides compositions comprising
cyclosporin (not a macromolecule) and containing BHA, BHT and PG
generally as antioxidants. Although no specific concentrations of
the antioxidants are given, the use of the compounds as
antioxidants suggests a level of no greater than 0.1% wt.
[0023] The alcohol may be present in the composition in an amount
of from 5 to 30% by weight, preferably from 10 to 20%.
[0024] The active macromolecular principles falling within the
scope of the invention include all molecules capable of having a
beneficial effect when absorbed into the human or animal body,
especially through the intestinal wall. The beneficial effect may
be, for example, therapeutic, cosmetic or preventative such as
prophylactic or contraceptive. The active macromolecular principles
can be of natural (biological), synthetic or semi-synthetic
origin.
[0025] Macromolecules are preferably defined as molecules having a
molecular weight of over 1000 Da, preferably over 2000 Da and most
preferably over 3000 Da. Examples of macromolecules, including
macromolecular active macromolecular principles, include:
[0026] 1. Polypeptides and proteins such as insulin; calcitonin;
human serum albumin; growth hormone; growth hormone releasing
factors; galanin; parathyroid hormone; peptide YY; oxyntomodulin;
blood clotting proteins such as kinogen, prothombin, fibrinogen,
Factor VII, Factor VIII of Factor IX; erythropoeitins and EPO
mimetics; colony stimulating factors including GCSF and GMCSF;
platelet-derived growth factors; epidermal growth factors;
fibroblast growth factors; transforming growth factors; GLP-1;
exendin; leptin; GAG; cytokines; insulin-like growth factors; bone-
and cartilage-inducing factors; neurotrophic factors; interleukins
including IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9,
IL-10, IL-11, IL-12; interferons including interferon gamma,
interferon.-1a, interferon alphas; TNF alpha; TNF beta; TGF-beta;
cholera toxin A and B fragments; E. coli enterotoxin A and B
fragments; secretin; enzymes including histone deacetylase,
superoxide dismutase, catalase, adenosine deaminase, thymidine
kinase, cytosine deaminase, proteases, lipases, carbohydrases,
nucleotidases, polymerases, kinases and phosphatases; transport or
binding proteins especially those which bind and/or transport a
vitamin, metal ion, amino acid or lipid or lipoprotein such as
cholesterol ester transfer protein, phospholipid transfer protein,
HDL binding protein; connective tissue proteins such as a collagen,
elastin or fibronectin; a muscle protein such as actin, myosin,
dystrophin, or mini-dystrophin; a neuronal, liver, cardiac, or
adipocyte protein; a cytotoxic protein; a cytochrome; a protein
which is able to cause replication, growth or differentiation of
cells; a signalling molecule such as an intra-cellular signalling
protein or an extracellular signalling protein (eg hormone);
trophic factors such as BDNF, CNTF,NGF, IGF, GMF, aFGF, bFGF, VEGF,
NT3, T3 and HARP; apolipoproteins; antibody molecules; receptors in
soluble form such as T-cell receptors and receptors for cytokines,
interferons or chemokines; proteins or peptides containing
antigenic epitopes and fragments; and derivatives, conjugates and
sequence variants of any of the above. These and other proteins may
be derived from human, plant, animal, bacterial or fungal sources,
and extracted either from natural sources, prepared as recombinants
by fermentation or chemically synthesised.
[0027] 2. Polynucleotides such as long-chain linear or circular
single-, double- or triple-stranded DNA, single-, double- or
triple-stranded RNA, oligonucleotides such as antisense DNA or RNA,
and analogues thereof including PNA and phosphothioate derivates.
In one embodiment it is preferred that the polynucleotides used in
the invention contain a CpG motif. The coding sequence of the
polynucleotide may encode a therapeutic product, in particular the
coding sequence may encode an extracellular protein (e.g. a
secreted protein); an intracellular protein (e.g. cytosolic,
nuclear or membrane protein); a protein present in the cell
membrane; a blood protein, such as a clotting protein (e.g.
kinogen, prothrombin, fibrinogen factor VII, factor VIII or factor
IX); an enzyme, such as a catabolic, anabolic gastro-intestinal,
metabolic (e.g. glycolysis or Krebs cycle), or a cell signalling
enzyme, an enzyme which breaks down or modifies lipids; fatty
acids, glycogen, amino acids, proteins, nucleotides,
polynucleotides (e.g. DNA or RNA) or carbohydrate (e.g. protease,
lipase or carbohydrase), or a protein modifying enzyme, such as an
enzyme that adds or takes chemical moieties from a protein (e.g. a
kinase or phosphatase); a transport or binding protein (e.g. which
binds and/or transports a vitamin, metal ion, amino acid or lipid,
such as cholesterol ester transfer protein, phospholipid transfer
protein or an HDL binding protein); a connective tissue protein
(e.g. a collagen, elastin or fibronectin); a muscle protein (e.g.
actin, myosin, dystrophin or mini-dystrophin); a neuronal, liver,
cardiac or adipocyte protein; a cytotoxic protein; a cytochrome; a
protein which is able to cause the replication, growth or
differentiation of cells; a protein which aids transcription or
translation of a gene or regulates transcription or translation
(e.g. a transcription factor or a protein that binds a
transcription factor or polymerase); a signalling molecule, such as
an intracellular or extracellular signalling molecule (e.g. a
hormone); an immune system protein such as an antibody, T cell
receptor, MHC molecule, cytokine (e.g IL-1, IL-2, IL-3, IL-4, IL-5,
IL-6, IL-7, IL-8, IL-9, IL-10, TNF-, TNF-, TGF-), an interferon
(e.g. IFN-, IFN-, IFN-), chemokine (e.g. MIP-1, MIP-1, RANTES), an
immune receptor (e.g. a receptor for a cytokine, interferon or
chemokine, such as a receptor for any of the above-mentioned
cytokines, interferons or chemokines) or a cell surface marker
(e.g. macrophage, T cell, B cell, NK cell or dendritic cell
surfacemarker)(eg. CD 1, 2, 3, 4, 5, 6, 7, 8, 16, 18, 19, 28, 40,
or 45; or a natural ligand thereof), a trophic factor (e.g. BDNF,
CNTF, NGF, IGF, GMF, aFGF, bFGF, VEGF, NT3, T5, HARP) or an
apolipoprotein; a tumour suppressor (e.g. p53, Rb, Rap1A, DCC or
k-rev); a suicide protein (thymidine kinase or cytosine deaminase);
or a gene repressor. The proteins and peptides encoded by the
polynucleotides useful in the invention may be immunogenic i.e.
contain an antigen specific to the activity of the protein against
which antibodies are generated by the immune system.
[0028] The polynucleotide may have control sequences operably
linked to the coding sequence. The control sequences may typically
be those of any eukaryote or of a virus which infects such
eukaryotes. The polynucleotide may comprise an origin of
replication.
[0029] The polynucleotides may be chemically modified. This may
enhance their resistance to nucleases or may enhance their ability
to enter cells. For example, phosphorothioate oligonucleotides may
be used. Other deoxynucleotide analogs include methylphosphonates,
phosphoramidates, phosphorodithioates, N3'P5'-phosphoramidates and
oligoribonucleotide phosphorothioates and their 2'-O-alkyl analogs
and 2'-O-methylribonucleotide methylphosphonates. Alternatively
mixed backbone oligonucleotides (MBOs) may be used. MBOs contain
segments of phosphothioate oligodeoxynucleotides and appropriately
placed segments of modified oligodeoxy- or oligoribonucleotides.
MBOs have segments of phosphorothioate linkages and other segments
of other modified oligonucleotides, such as methylphosphonate,
which is non-ionic, and very resistant to nucleases or
2'-O-alkyloligoribonucleotides.
[0030] The polynucleotide suitable for use in the invention is
preferably in a form in which it is substantially free of or
associated with cells or with cellular, prokaryotic, eukaryotic,
nuclear, chromatin, histone or protein material. It may be in
substantially isolated form, or it may be in substantially purified
form, in which case it will generally comprise more than 90%, e.g.
(more than or at least) 95%, 98% or 99% of the polynucleotide or
dry mass in the preparation. Thus the polynucleotide may be in the
form of `naked DNA`.
[0031] 3. Polysaccharides such as heparin, low-molecular weight
heparin, polymannose, cyclodextrins and lipopolysaccharide.
[0032] 4. Any or all of the above either separately or in
combination with each other (for example in the form of a
heteroconjugate), or with additional agents.
[0033] In preferred embodiments of the invention the active
macromolecular principle to be absorbed is selected from
calcitonins, insulin, low molecular weight heparin, erythropoeitin,
colony stimulating factor, including granulocyte colony stimulating
factor (GCSF) and granulocyte monocyte colony stimulating factor
(GMCSF), interferons, C-peptide, glucagons-like protein 1 (GLP-1),
human growth hormone and parathyroid hormone and analogues and
fragments thereof.
[0034] In the compositions of the invention, the aromatic alcohol
absorption enhancer is present in an amount (by weight) greater
than or equal to that of the active macromolecular principle. This
provides an effective concentration of aromatic alcohol absorption
enhancer at the intestinal cell barrier layer (intestinal wall) so
as to cause enhanced absorption in the co-presence of a suitable
amount of the active macromolecular principle which, when absorbed,
will exert its normal beneficial effect. The amounts of the
aromatic alcohol absorption enhancer and active macromolecular
principle are readily selected on the basis of the amount (for
example, blood concentration level) of the active macromolecular
principle concerned which is necessary for therapeutic efficacy.
The, weight ratio of aromatic alcohol absorption enhancer to active
macromolecular principle in the mixture contained in the capsule is
suitably at least 1:1, preferably at least 5:1, for example from
1:1 to 100:1, preferably from 3:1 to 50:1, most preferably from 5:1
to 20:1.
[0035] The ratio of biguanide to aromatic alcohol absorption
enhancer is suitably at least 2:1 by weight, preferably from 2:1 to
10:1, and most preferably from 2:1 to 5:1.
[0036] The absolute amount of the active macromolecular principle
would be selected on the basis of the dosage of the substance
required to exert the normal beneficial effect with respect to the
dosage regimen used and the patient concerned. Determination of
these amounts falls within the mantle of the practitioner of the
invention.
[0037] In the composition for oral administration it is preferred
that the contents of the capsule comprises a suitable amount of the
active macromolecular principle to achieve its normal therapeutic
effect. For example, the composition may contain from 0.05 to 50%,
preferably from 0.1 to 25%, more preferably from 0.1 to 10% by
weight of the active macromolecular principle based on the weight
of the capsule contents (not including the capsule itself).
[0038] The composition of the invention may further comprise one or
more other absorption enhancer compounds, for example, medium chain
fatty acids and medium chain monoglycerides.
[0039] The composition of the invention may optionally further
comprise any conventional additive used in the formulation of
pharmaceutical products including, for example, anti-oxidants,
anti-microbials, suspending agents, fillers, diluents,
disintegrants, swelling agents, viscosity regulators, plasticisers
and acidity regulators (particularly those adjusting the intestinal
milieu to between 7 and 7.5) and protease inhibitors such as
aprotinin, soybean trypsin inhibitor or gabexate mesylate.
[0040] The composition of the invention may optionally further
comprise additional active principles which may enhance the desired
action of the composition in a synergistic fashion. For example,
where the active macromolecular principle is insulin, the
composition may also comprise an insulin sensitiser capable of
increasing the body's response to the insulin absorbed. Examples of
sensitisers which could be employed in this fashion are
troglitazone, pioglitazone, rosiglitazone and other members of the
glitazone class of molecules.
[0041] In the composition of the invention where the mixture is
contained in a capsule or tablet that comprises the aromatic
alcohol absorption enhancer and active macromolecular principle,
the formulation is preferably substantially anhydrous. In more
preferred embodiments of the invention the entire composition is
substantially anhydrous. Substantially anhydrous in the context of
this invention means less than 5%, preferably less than 1% and more
preferably less than 0.5% water by weight of the mixture.
[0042] The compositions of the invention can, depending on the
active macromolecular principle used therein, be used in the
treatment of a variety of conditions and diseases of the human or
animal body by therapy or, alternately, can be used to introduce
macromolecules essential for the diagnosis of diseases and
conditions within the human or animal body. The compositions of the
invention are preferably pharmaceutical or cosmetic
compositions.
[0043] The pharmaceutical compositions of the invention are
particularly useful in the treatment of diabetes when the
composition may comprise insulin, C-peptide or GLP-1 or a mixture
thereof as active principle, in the treatment of osteoporosis when
the composition may comprise calcitonin or PTH or a mixture
thereof, in the treatment of osteoarthritis, when the composition
may comprise calcitonin, in the treatment of obesity, where the
composition may comprise peptide YY or oxyntomodulin or a mixture
thereof, or in the treatment of cancer, where the composition may
comprise erythropoetin, GCSF, GMCSF or a mixture thereof.
[0044] In the compositions of the invention the mixture contained
in the capsule may be a liquid, semi-solid or gel, which is either
in the form of a solution or a microparticulate dispersion. That is
to say the active macromolecular principle(s) for absorption are
incorporated into the formulation either in the form of a solution
or as a microparticulate dispersion. Alternatively, the composition
may be in the form of a solid.
[0045] The compositions of the invention are suitably produced by
preparing a substantially anhydrous mixture of the active
macromolecular principle and the aromatic alcohol absorption
enhancer and then optionally filling uncoated capsules with the
mixture and optionally coating them with an appropriate polymer
mixture to achieve the desired permeability properties.
[0046] Depending on the nature of additional excipients employed,
the pharmaceutical composition of the invention may be in liquid,
solid, semi-solid or gel form. The pharmaceutical composition of
the invention is suitable for administration via any route giving
access to different mucosal tissues such as buccal and sublingual
mucosa, the nasal palate, the lungs, the rectum, the intestinal
tract (including the large and small intestines) and the vagina. In
the case of liquid, semi-solid or gel formulations, these may be
either anhydrous or aqueous.
[0047] Where the intended site of action of the composition of the
invention is the intestine, it is desirable that the composition is
enclosed within an enteric coating which can withstand the stomach,
so that the components of the formulation remain together,
undiluted and in close association until they reach the tissues of
the small intestine or colon. Such formulations will suitably be
anhydrous. Compositions in liquid form will suitably be
administered as enteric-coated capsules, while solid formulations
may be administered either within enteric-coated capsules, or in
tablet form.
[0048] The enteric coating is chosen appropriately to withstand the
natural condition of the stomach and to become permeable at the
desired location in the intestine. This is preferably determined by
the pH conditions which modulate along the length of the intestine.
Where the site of action is the small intestine, it is preferred
that the enteric coating becomes permeable and releases its
contents at a pH from 3 to 7, preferably from 5.5 to 7, more
preferably from 5.5 to 6.5. Where the intended site of action is
the colon, it is preferred that the enteric coating becomes
permeable and releases its contents at a pH of 6.8 or above.
[0049] Suitable enteric coatings are well known in the art and
include polymethacrylates such as those selected from the L and S
series of Eudragits in particular Eudragits L12.5P, L12.5, L100,
L100-55, L30 D-55, S12.5P, S12.5 and S100. Selection of an
appropriate coating for the capsule, which is preferably a gelatine
capsule, can readily be made by the person skilled in the art based
on their knowledge and the available literature supporting the
Eudragit products.
[0050] Where the intended site of action is the nasal mucosa, the
formulation may be in the form of an aqueous solution or as a dry
powder, which can be administered as a spray.
[0051] Where the intended site of action is the rectum, an
appropriate method of administration is as an anhydrous liquid or
solid enclosed within a capsular shell, or incorporated into the
matrix of an erodible suppository.
[0052] For vaginal application, adminstration of the formulation in
gel form is also
[0053] The following Examples serve to illustrate the present
invention and should not be construed as limiting.
EXAMPLES
Example 1
Preparation of a Formulation Containing Metformin and Propyl
Gallate
[0054] To 1 g metformin 1.83 g of distilled water is added. The
mixture is then warmed up at approximately 60.degree. C. with
gentle shaking until all solid has dissolved. To the clear solution
250 mg of propyl gallate is added. The mixture is then warmed up at
approximately 60.degree. C. with gentle shaking until all solid has
dissolved. The pH is adjusted to 5.5 by addition of approximately
25 ul of sodium hydroxide. A homogenous solution is obtained which
remains clear at room temperature.
Example 2
Preparation of a Formulation Containing Phenformin and Propyl
Gallate
[0055] A phenformin/propyl gallate formulation is prepared as
described in example 1 except that metformin is replaced with
phenformin. A homogenous solution is obtained which remains clear
at room temperature.
EXAMPLE 3
Preparation of a Formulation Containing Chlorhexidine Digluconate
and Propyl Gallate
[0056] Chlorhexidine digluconate solution at 200 mg/ml is diluted
2.times. with distilled water giving solution of 10 mg/ml. To 10 ml
of this solution 250 mg of propyl gallate is added. The mixture is
then warmed up at approximately 60.degree. C. with gentle shaking
until all solid has dissolved. The pH is adjusted to 5.5 by
addition of approximately 25 ul of 1M sodium hydroxide. A
homogenous solution is obtained which remains clear at room
temperature.
EXAMPLE 4
Preparation of a Formulation Containing Metformin and Butylated
Hydroxy Toluene (BHT)
[0057] To 400 mg metformin 4.0 g of distilled water is added. The
mixture is then warmed up at approximately 70.degree. C. with
gentle shaking until all solid has dissolved. To the clear solution
40 mg of BHT is added. The mixture is then warmed up at
approximately 70.degree. C. with gentle shaking until all solid has
dissolved. A homogenous solution is obtained which remains clear at
room temperature.
EXAMPLE 5
Preparation of a Formulation Containing Phenformin and Butylated
Hydroxy Toluene (BHT)
[0058] To 400 mg phenformin 4.0 g of distilled water is added. The
mixture is then warmed up at approximately 70.degree. C. with
gentle shaking until all solid has dissolved. To the clear solution
100 mg of BHT is added. The mixture is then warmed up at
approximately 70.degree. C. with gentle shaking until all solid has
dissolved. A homogenous solution is obtained which remains clear at
room temperature.
EXAMPLE 6
Preparation of a Formulation Containing Phenformin and Butylated
Hydroxy Anisole (BHA)
[0059] The preparation method for the Metformin/BHT formulation is
as described in example 5 except that BHT is replaced with BHA. A
homogenous solution is obtained which remains clear at room
temperature.
EXAMPLE 7
Preparation of a Formulation Containing Metformin, Propyl Gallate
and Insulin
[0060] The metformin/propyl gallate solution is prepared as
described in Example 1. The obtained solution is then cooled down
to 37.degree. C. and 28.1 mg of insulin is added. The mixture is
then incubated at 37.degree. C. with mixing till the insulin is
completely dissolved. The solution is then frozen rapidly at
-20.degree. C., incubated further at -20.degree. C. for 1 hour and
then freeze-dried overnight, by exposing to vacuum of 1 mbar. The
dry powder cake is then passed through a sieve so a fine powder is
obtained.
EXAMPLE 8
Dissolution of a Formulation Containing Metformin, Propyl Gallate
and Insulin
[0061] 244mg of the Metformin/PG/Insulin powder is weighed into an
8 ml vial that is then transferred into a 37.degree. C. water bath.
1 ml of simulated intestinal fluid pH 5.5 pre-warmed to 37.degree.
C. is introduced to the sample that is then incubated at 37.degree.
C. The powder dissolves within 10 minutes. 100 ul of the solution
is transferred to a well of the microplate and the absorbance is
measured at 620 nm and 492 nm. The optical density of the solutions
is similar to that for simulated intestinal fluid alone,
demonstrating that the solution is clear and free from particles,
and no scattering is observed.
TABLE-US-00001 Sample Absorbance at 492 nm Absorbance at 620 nm
Powder 0.064 0.038 Intestinal fluid 0.038 0.043
EXAMPLE 9
In Vivo Efficacy of Metformin Hydrochloride/Propyl Gallate/Insulin
Mixture in Juvenile Pigs
[0062] Formulations as prepared in example 7 are mixed with
swelling agent and glidant, and the dry powder is filled into
capsules, each having components in the proportions shown below.
Formulations containing chenodeoxycholate instead of metformin, and
salmon calcintonin instead of insulin are prepared for comparison.
All formulations have identical amounts of propyl gallate per
capsule.
TABLE-US-00002 41B 90A 88E Insulin 3.75 mg 3.75 mg Calcitonin 1.00
mg Metformin 133.4 mg 133.4 mg Chenodeoxycholate 70.60 mg Propyl
gallate 33.35 mg 33.35 mg 33.35 mg Sodium starch 9.69 mg 15.15 mg
15.15 mg glycolate Fumed silica 1.08 mg 1.67 mg 1.67 mg Soybean
trypsin 10.00 mg inhibitor
[0063] The capsules are administered in capsules via a stoma into
the jejunum of eight juvenile pigs (each 40 kg weight). Blood
glucose levels are measured at intervals over a six hour period and
mean change in AUC of plasma glucose is calculated in h.mmol/l. As
can be seen from the summary of data below, the formulation
containing the metformin/propyl gallate combination displays
efficacy equal to or greater than that of chenodeoxycholate/propyl
gallate.
TABLE-US-00003 AUC of change in plasma glucose after 6 hours (h
mmol/l) 41B Insulin/Cheno/PG -2.25 90A Insulin/metformin/PG -4.02
88E Calcitonin/metformin/PG + SBTI -0.21 (-ve control)
EXAMPLE 10
In Vivo Efficacy of Metformin Hydrochloride/Propyl
Gallate/Calcitonin Mixture in Juvenile Pigs
[0064] Formulations as prepared in example 9 (containing 6000 iu
salmon calcitonin, 133 mg metformin hydrochloride and 33 mg propyl
gallate or 6000 iu salmon calcitonin and metformin hydrochloride
alone) are administered as dry powders inside capsules via a stoma
into the jejunum of eight juvenile pigs (each 40 kg weight).
Compositions of the contents of each capsule are shown in the table
below. Formulations containing chenodeoxycholate are included for
comparison.
TABLE-US-00004 90A 81A 81B 82A 82B 84A 84B Calcitonin 1 mg 1 mg 1
mg 1 mg 1 mg 1 mg Insulin 3.75 mg Metformin 133.4 mg 133.4 mg 133.4
mg 133.4 mg 133.4 mg Chenodeoxycholate 70.6 mg 70.6 mg Propyl
gallate 33.35 mg 33.35 mg 33.35 mg 33.35 mg 33.35 mg Sodium starch
glycolate 15.15 mg 9.45 mg 9.45 mg 15.15 mg 15.15 mg 15.15 mg 15.15
mg Fumed silica 1.67 mg 1.05 mg 1.05 mg 1.67 mg 1.67 mg 1.67 mg
1.67 mg Soybean trypsin inhibitor 10.00 mg 10.00 mg Aprotinin 10.00
mg 10.00 mg
[0065] Blood calcium levels are measured at intervals over a six
hour period and mean change in AUC of plasma calcium is calculated
in h.mmol/l. As can be seen from the summary of data below, calcium
levels are reduced below the baseline as a result of introduction
of calcitonin into the bloodstream from the intestine. The
metformin/PG combination is most efficacious, more so than the
metformin in the absence of PG, indicating that the absorbtion
enhancing effect is not due to metformin itself, but is the result
of action of the propyl gallate, whose dissolution in aqueous media
in the gut is brought about by the presence of the metformin.
Further enhancement of activity can be brought about by inclusion
of protease inhibitors in the formulations.
TABLE-US-00005 AUC of change in plasma calcium after 6 hour (h
mmol/l) 90A Insulin/metformin/PG (-ve control) -0.51 81A
Calcitonin/chenodeoxycholate/PG -0.89 81B
Calcitonin/chenodeoxycholate/PG + -1.43 SBTI 82A
Calcitonin/Metformin/PG -1.12 82B Calcitonin/Metformin/PG +
aprotinin -2.04 84A Calcitonin/Metformin - no PG -0.65 84B
Calcitonin/Metformin + aprotinin - -0.68 no PG
EXAMPLE 11
In Vivo Efficacy of Metformin Hydrochloride/Propyl
Gallate/Calcitonin Mixture in Juvenile Pigs
[0066] A formulation as prepared in example 9 (each capsule
containing 4 mg parathyroid hormone, 133.4 mg metformin
hydrochloride, 33.35 mg propyl gallate, 15.15 mg sodium starch
glycolate, 1.65 mg fumed silica and 10.00 mg soybean trypsin
inhibitor) is administered via a stoma into the jejunum of eight
juvenile pigs (each 40 kg weight). Blood calcium levels are
measured at intervals over a six hour period. As can be seen from
the data below, calcium levels are changed from the baseline as a
result of introduction of PTH into the bloodstream from the
intestine.
TABLE-US-00006 Peak-to-trough difference in plasma calcium (mmol/l)
PTH solution in buffered saline 0.31 s.c (0.4 mg) 88B PTH in
Metformin/PG formulation 0.27 i.j. (4.0 mg)
* * * * *