Note: Descriptions are shown in the official language in which they were submitted.
W093/06~54 2 1 2 0 7 ~ ~ PCT/~P92/02321
PHARMACEUTICAL COMPOSITIONS COMPRISING A CALCITOI~ ;. A CiLYCYR-
RHIZINATE AS ABSORPTION ENHANCER AND BENZYL
The present invention relates to novel pharmaceutical
compositions containing calcitonins, and to a novel method of
enhancing the absorption of a calcitonin across a mucosal
- membrane.
The calcitonins are a class of pharmacologically active
peptides, of both natural and synthetic origin, which contain
o approximately thirty two amino acids, and which have the
ability to regulate serum calcium levels.
Various calcitonins, including e.g. natural human, salmon and
~ eel calcitonins and the synthetic eel calcitonin analogue
elcatonin are now commercially available and commonly
employed,~ e.g. in the treatment of Paget's disease, Sudeck's
disease and osteoporosis.
A considerable and well known problem with the administration
~of peptides is that they are suscept~ble to rapid acid and
enzyme-induced degra~ation when administered orally. For this
reason, parenteral administration has been, hitherto, the
most widely used means of administration and, in the case of
peptides of higher molecular weight, such as the calcitonins,
has been the only significant effective means o~
administration.
It is widely recognised that administration by injection can
be bot~ inconvenient and unpleasant for the patient,
particularly when the administration has to be repeated at
regular intervals for long periods, e.g. in the treatment of
post-menopausal osteoporosis with calcitonins. Thus, there
has been growing interest in the administration of peptides
by more acceptable non-invasive alternative routes, for
example in the form of sublingual tablets, suppositories,
intrapulmonary powders, intranasal drops, sprays, powders,
gels, ointments and inserts.
''~
2 12 ~ 7 ~ 5 PCT~EP92/0~21
- 2 -
A significant problem with many peptides, particularly those ;;
of higher molecular weights, is that they are only poorly
absorbed across biological membranes, e.g. mucosal membranes,
and thus the bioavailability of the peptide is often very
s low. Considerable research has therefore been carried out in
order to find methods of improving the trans-epithelial
absorption of peptides. One approach is to use an adjuvant or
absorption enhancer and there are numerous published reports ~-
of compounds which are claimed to have peptide absorption-
enhancing properties.
Thus, for example, choline esters ~EP 214898), acyl ~-~
carnitines (EP 215697), aldoses and glucosamines (Japanese
Pat. Appl. No. 61 126034), ascorbates and salicylates
(EP 37943), alpha-cyclodextrin (EP 0094157), pyroglutamate
estérs (EP 173990), chelating agents ~US 4,476,116) ethanol,
benzyl alcohol and polyethylene glycol 400 (EP 371010) have
been proposed as absorption enhancers.
20 ~There are many published reports that surfactants can enhance
the absorption of polypeptides, see for example EP 115627
(Armour), GB 2,127,689 (Sandoz), US 4,548,922 ~Carey et ~l)
and Hirai ~ al., Int~ J. ~h~Lm_, 2, 165-184, 1981. However,
a recognised problem with surfactant absorption promoters is
2s that they can cause irritation and histolesion at the site of
administration. These problems become of great importance
when the peptide is administered regularly over a prolonged
period~
30 The present applicants have previously found that ~ ;
glycyrrhizinic acid and its salts are excellent absorption
promoters for calcitonins and do not give rise to the above-
mentioned problems of local toxicity and irritation. i-
Compositions comprising a calcitonin and a gl~cyrrhizinate
are described in our EPA 327756, which includes both liquid
and solid formulations. Liquid formulations conventionally
contain a preservative and EPA 327756 refers to the use of -`
W093/06854 212 0 7 5 5 pcr/Eps2/o232l ~ ~
- 3 - ~:
alkyl p-hydroxybenzoates (parabens) such as methyl and propyl
p-hydroxybenzoate as suitabl preservatives.
However, it has subsequently been shown that the
antibacterial and preservative actions of the parabens are
reduced by the glycyrrhizinate component of the formulation.
In addition it would be desirable to increase the absorption ::.
of calcitonins still further.
o We have now surprisingly foun~ that the inclusion of benzyl
alcohol in a composition comprising a calcitonin and a
glycyrrhizinate not only gives rise to a preservative action
which is not diminished by the glycyrrhizinate, but also
enhances the absorption of the calcitonin in a synergistic
manner. Thus the use of a glycyrrhizinate in combination
with benzyl alcohol increases t~e transmucosal absorption of
a calcitonin by more than the sum of the respective effects
of benzyl alcohol and glycyrrhizinate alone.
20 ~ In a first aspect, therefore, the present invention provides
pharmaceutical compositions comprising a calcitonin; an
effective amount of an absorption enhancer which is a
glycyrrhizinate; an effective amount of benzyl alcohol and a
pharmaceutically acceptable carrier.
The present invention also provides a method of enhancing the
absorption of a calcitonin across a mucosal membrane, which
method comprises co-administering with the calcitonin an
effective amount of an absorption enhancer which is a
g~ycyrr~izinate, and an effective amount of benzyl alcohol.
Whilst preservatives are generally only used in liquid
formulations, absorption enhancers are required in both
liquid and solid formulations of calcitonins, and hence the
precent invention includes within its scope both solid and
liquid compositions.
W093/068~ 212 0 7 S 5 4 PCT/EP92/0~21 -~
- .-
The term glycyrrhizinate as used herein is intended to mean
both glycyrrhizinic acid and its carboxylate salts. -~
Particular glycyrrhizinate salts are ammonium glycyrrhizinate
and the alkali metal salts e.g. sodium glycyrrhizinate and
potassium glycyrrhizinate. A preferred salt is ammonium
glycyrrhizinate.
The term calcitonin as used herein is intended to refer to
that class of pharmacologically active polypeptides including ~`
o not only naturally occurring calcitonins but also various
derivatives and analogues thereof, e.g. in which one or more
of the amino acid residues or sequences naturally present is
omitted, replaced, reversed or otherwise derivatised or in ~-
which the N- or C-terminal is modified.
The general term calcitonin, as used hereinafter, is intended ~:
to mean all such calcitonins whether naturally occurring or
synthetic~
20 ~Examples of naturally occurring calcitonins include: human
calcitonin, Chemical Abstract Service Registry Number (CAS
RN) = 21215 62-3, which has the structure:
. .
~-Cys-Gly-Asn-Leu-Ser-Thr-Cys-Met-Leu-Gly
-Thr-Tyr-Thr-Gln-Asp-Phe-ASn-Lys-Phe-His-
-Thr-Phe-Pro-Gln-Thr-Ala-Ile-Gly-Val-Gly r
-Pla-Pro-NH2;
rat calcitonin SCAS RN = 11118-25-5) which has the
structure:
\
H-Cys-Gly-Asn-Leu-Ser-Thr-Cys-Met-Leu-Gly-
-Thr-Tyr-Thr-Gln-Asp-Leu-Asn-Lys-Phe-His-
-Thr-Phe-Pro-Gln-Thr-Ser-Ile-Gly-Val-Gly-
-Ala-Pro-NH2;
W093/068~ 2 ~ 2 ~ 7 ~ ~ PCT/EP92/02321
salmon calcitonin (CAS RN = 47931-85-1) which has the
structure:
I ~
H-Cys-Ser-Asn Leu-Ser-Thr-Cys-Val-Leu-Gly-
-Lys-Leu-Ser-Gln-Glu-Leu-His-Lys-Leu-Gln-
-Thr-Tyr-Pro-Arg-Thr-Asn-Thr-Gly-Ser-Gly-
-Thr-Pro NH2;
eel calcitonin ~CAS RN = 57014-02-5) which has the structure:
I
H-Cys-Ser Asn-Leu-Ser-Thr-Cys-Val-Leu-Gly-
-Lys-Leu-Ser-Gln-Glu-Leu-His-Lys-Leu-Gln-
-Thr-Tyr-Pro-Arg-Thr-A~p-Val-Gly-Ala-Gly-
-Thr-Pro-NH2;
reduced chicken calcitonin I (CAS RN = 96157-98-1) which
`has the structure: :
20 - ``
H-Cys-Ala-Ser-Leu-Ser-Thr-Cys-Val-Leu-Gly-
-Lys-Leu-Ser-Gln-Glu-Leu-His-Lys-Leu-Gln- ~
-Thr-Tyr-Pro-Arg-Thr-Asp-Val-Gly-Ala-Gly- ~-
-Thr-Pro-NH2;
chicken ~alcitonin II (CAS ~N - 103468-65-1) which has the
structure:
~-gamma-Glu-Cys-Gly-OH H-gamma-Glu-Cys-Gly-5H
H-Cys-Ala-Ser-Leu-Ser-Thr-Cys-Val-Leu-Gly-
-Lys-Leu-Ser-Gln-Glu-Leu-His-Lys-Leu-Gln-
-Thr-Tyr-Pro-Arg-Thr-Asp-Val-Gly-Ala-Gly-
-Thx-Pro-NH2;
ox calcitonin (CAS RN = 26112-29-8) which has the structure:
W093/068~ 2 1 2 0 ! ~i .i PCT/EPg2/0~21
-- 6 --
H-Cys-Ser-Asn-Leu-Ser-Thr-Cys-Val Leu-Ser-
-Ala-Tyr-Trp-Lys-Asp-Leu-Asn-Asn~Tyr-His-
~Arg-Phe-Ser-Gly-Met-Gly-Phe-Gly-Pro-Glu-
-Thr-Pro-NH2;
pig calcitonin (CAS RN = 12321-44-7) which has the structure:
H-Cys-Ser-Asn-Leu-Ser-Thr-Cys-Val-Leu-Ser-
-Ala-Tyr-Trp-Arg-Asn-Leu-Asn-Asn-Phe-H~s-
-Arg-Phe-Ser-Gly-Met-Gly-Phe-Gly-Pro-Glu-
-Thr-Pro-NH2; and
:
sheep calcitonin (CAS RN =.40988-57-6) which has the
structure:
I` - '' - ~ ,.
20 ~H-Cys-Ser-Asn-Leu-Ser-Thr-Cys-Val-Leu-Ser-
-Ala-Tyr-Trp-Lys-Asp-Leu-Asn-Asn-Tyr-His- -
-Arg-Tyr-Ser-Gly-Met-Gly-Phe-Gly-Pro-Glu-
-Thr-Pro-NH2 .
Examples of calcitonins wherein one or more amino acids -~
have been omitted are the des-[Ser2, Tyr2~ Gly8-calcitonins
described in US 4,597,900 and the des-[Tyr22]-salmon
calcitonin described in US 4~304,692. ~-
Examples o`f calcitonins wherein the naturally occurring
sequence has been modified include the 1,7-dicarba~ ~
calcitonins such as eel 1,7-dicarbacalcitonin (elcatanin CAS :
RN = 60731-46-6) which has the structure:
(CH2)s 1 ~1
CO-Ser-Asn-Leu-Ser-Thr-NH-CH-C0-Val-Leu-Gly-
-Lys-Leu-Ser-Gln-Glu-Leu-His-Lys-Leu-Gln-
-Thr-Tyr-Pro-Arg-Thr-Asp-Val-Gly-Ala-Gly-
-Thr-Pro-NH2;
W093/068~ 2 1 2 ~ 7 ~ ~ PCT/EP92/02321
salmon l,7-dicarbacalcitonin (CAS RN = 60864-37-1) which has
the structure:
5 1 _ (CH2)5
I
CO-Ser-Asn-Leu-Ser-Thr-NH-CH-CO-Val-Leu-Gly-
-Lys-Leu-Ser-Gln-Glu-Leu-His-Lys-Leu-Gln- ~-
-Thr-Tyr-Pro-Arg-Thr-Asn-Thr-Gly-Ser-Gly-
-Thr-Pro-NH2; and
human 1,7-dicarbacalcitonin (CAS RN - 66811-56-1) which has
the structure~
15 1 (CH2)5
~ .
CO-Gly-Asn-Leu-Ser-Thr-NH-CH-CO-Met-Leu-Gly-
-Thr-Tyr-Thr-Gln-Asp-Phe-Asn-Lys-Phe-His-
-Thr-Phe-Pro-Gln-Thr-Ala-Ile-Gly-Val-Gly~
20 _ -Ala-Pro-NH2. ~
In the context of the present invention, a particularly -`
preferred calcitonin is elcatonin ~CAS RN = 60731-46- 6). The
preparation and properties of elcatonin and related 1,7-
25 dicarbacalcitonins are described in British Patent Number ::
1,516,947 (Toyo Jozo~.
Another preferred calcitonin is naturally occurring eel
calcitonin (CAS RN = 57014-02-5). The preparation and
properties of eel calcitonin are described in US 3,988,309
(Matsuda ~
The compositions of the present invention suitably can be
administered by methods known in the art for transmucosal and
transdermal delivery of pharmacologicall~ active substances.
The compositions can be administered to, for example, the
nasal, sublinguat, buccal, rectal, vaginal and colonic mucosa
and to the skin. They can take the form of drops, aerosols,
w093/06854 2 1 2 0 7 ~ rj PCT/EP92~02321
-- 8
tablets, capsules, powders, gels, ointments, inserts,
suppositories, pessaries, patches and membranes. The
compositions can also take the form of enterically coated
solid oral compositions as described in, for example, EP
127535 (Hadassah Medical Organisation). The compositions for
sublingual and buccal administration can also take the form ~-
of wafers as described in PCT/GB91/00651. Such wafers are
forméd substantially from starch, and suitably have a
thickness of from 0.3 to 1.0 mm.
Particular compositions are those intended for administration
to the nasal, buccal, sublingual, rectal and vaginal mucosa.
When the composition is intended for delivery to the nasal
mucosa, particular dosage forms are solutions, aerosols,
drops, gels and powders.
Particular dosage forms for buccal and sublingual
administration are gels, suspensions, tablets, patches,
20 _powders, ointments, solutions, aerosols and wafers. i~
Aerosol formulations typically comprise a solution or fine
suspension of the active substance in a physiologically
acceptable aqueous or non-aqueous solvent and are usually
presented in single or multidose quantities in a sealed
container. The sealed container can take thei form of a
cartridge or refill for use with an atomising device, or it
can take the form of a unitary dispensing device such as a
single dose nasal inhaler ~see French Patent ~pplication
FR 2578426) or an aerosol dispenser fitted with a metering
valve and which is intended for disposal once the contents of
the container have been exhausted. Where the dosage form
comprises an aerosol dispenser, it will contain a propellant
which can be a compressed gas such as compressed air or an
organic propellant such as a fluorochlorohydrocarbon. Such
aerosol dispensers are well known in the art. The ~erosol
dosage forms can also take the form of a pump-atomiser and
such forms are also well known in the art. The atomising or
W093/~ 2 1 2 ~ 7 5 5 PCT/EP92tO2321
dispensing devices for dispensing aerosol sprays typically
are designed to dispense particles of a size greater than 10
micrometres. In order to ensure that significant quantities
of the composition remain in contact with the oral or nasal
mucosa, and are not inhaled, the particles suitably are
approximately 10-160 micrometres in size.
When the composition is intended to be administered as a
liquid spray, the viscosity of the liquid composition can be
lo adjusted as necessary according to known methods to ensure
that ~he composition is sprayable.
When the composition is intended for application to the
rectal and vaginal mucosa particular dosage forms include
pessaries, suppositiories, solutions, foams, suspensions,
gels, ointments, tablets and soft gelatin capsules.
Compositions for rectal or ~Jaginal administration are
generally presented as a solid suppository or a semisolid or
20 _ liquid formulation filled into a soft gelatin capsule. It
will be appreciated therefore that the excipients for use in
such suppository or capsule formulations will be selected and
if necessary admixed to give a formulation of the desired
consistency at room and body temperatures. Thus, the
suppository base or carrier may for example comprise one or
more components selected from an oil, a fat, a polyglycolysed
glyceride and a polyethylene glycol. The oil and/or fat
preferably co~prises one or more triglycerides as the main
component, such as coconut oil, fractionated coconut oil
(e.g. Miglyol? palm kernel oil, palm fat, cocoa butter or
lard. Examples of hard fat suppository bases include
Witepsol and Suppocire. A saturated or unsaturated
polyglycolysed glyceride may be for example a saturated
polyglycolysed glyceride consisting of Cg_l~ glycerides and
polyethylene glycol esters such as are available under the
trade name Gelucire e.g. Gelucire 35/10, 37~02 or 44/14; a
saturated polyglycolysed Cg_Clo glyceride such as that
available under the trade name Labrasol; or an unsaturated
W093/06854 2 ~ 2 ~ 7 ~ ~ PCT/EP92tO~21
-- 10 --
polyglycolysed glyceride consisting of C16-C20 glycerides and
polyethylene glycol esters such as those available under the
trade name Labrafils e.g. Labrafil WL 2609 BS or M 2125 CS.
For use in a capsule formulation the polyethylene glycol
s component is preferably liquid at room temperature such as
polyethylene glycol-200, 300, 400 or 600, whereas for a solid
suppository a polyethylene glycol of higher molecular weight
is preferred. The relative proportions of the excipients
will of course depend in~L alia on the consistency of the
0 formulation required.
: ~.
Compositions containing a polyglycolysed glyceride optionally ;
with a polethylene glycol are preferred. Such compositions
can also be adapted for oral administration e.g. in hard or
soft gelatin capsules, which are preferably enterically :
coated.
.
When the composition is enterically coated and is intended
for oral administration, typically it can take the form of a
20 ~tablet or capsule coated with a coating agent which ensures
passage of the calcitonin through the stomach and its
subsequent release preferably in the colon. Suitable coating
agents include anionic polymers such as acrylic
acid/methacrylic acid ester copolymers (e.g. Eudragit S).
The solvents or liquid carriers used in the present
formulations are preferably aqueous but can also be chosen
from the physiologically acceptable non-aqueous solvents.
Examples of non-aqueous solvents or carriers are alcohols,
particularly polyhydroxy alcohols such as propylene glycol
and glycerol, and vegetable and mineral oils. Such non-
aqueous solvents or carriers can be added in various
concentrations to water to form solutions, oil-in-water
emulsions and water-in-oil emulsions. The solvent preferably
is water.
:.
In addition to a solvent or carrier, the liquid formulations -~
of the present invention can contain excipients such as
W093/068~ PCT/EP92/02321
219~7S5
antioxidants, stabilisers, preservative~, agents for
adjusting viscosity (e.g. Carbapol, Keltrol or cellulose
derivatives), agents for adjusting tonicity (e.g. sodlum
chloride, glycine or mannitol), and buffering agents. If
desired a further preservative eg. parabens may be used in
~ddition to ben~yl alcohol, bllt in general this is not
necessary.
.~ .
The compositions can also contain a protease inhibitor,
o preferably a non-surfactant protease inhibitor, for example
as described in EP 127535.
In~general, the above-mentioned compositions can be made
according to well known pharmaceutical procedures, see for
example Remington's Pharmaceutical Sciences, 17th Edition,
Mack Publishing Company, 1985. Soft gelatin capsules may be
prepared for example as described in WO 84~03417 or
EPA 122463. Wafer formulations may be prepared for example
as described in PCT/GB91/006510 Thus for example the active
20~ ingredient may be incorporated into the wafer mix prior to
forming the wafer, or applied to the wafer in the form of a
layer or a spray.
Th~ compositions of the present invention can be used in ~he
2s treatment of diseases such as PacJet ' s disease (osteitis
deformans)~ osteoporosis, including post-menopausal
osteoporosis; Sudeck's disease and various hypercalcaemic
conditions (see, for example, the Physician's Desk Reference,
42nd Edition, 1988, pages 1796 and 17973.
The compositions will be administered to the patient in
dosages which contain an amount of calcitonin effective to -~
treat the disease in question.
3s The quantity of pharmacologically active su~stance in a unit
dose of the compositions of the present invention will vary
according to the potency of the calcitonin and the nature of
the composition. However, in general, a unit dose of a `.
W093/06854 2 ~ 2 Q 7 5 ~ PCT/EP92/02321
- 12 -
composition intended for human use typically contains between
l and 400 International Units tI.U.) of a caloitonin. For
elcatonin, a unit dose preferably contains from 5 to 200 I.U.
A typical dosage regimen for elcatonin is from 5 to 200 I.U.
s per day which may be administered in a single dose or in
divided doses, f or example on consecutive or on alternate
days.
The term ~'International Unit" refers to the appropriate
International Reference Preparation (I.R.Pu) of human, salmon
or porcine calcitonins, or elcatonin, established by the
National Institute for Biological Standards and Control,
Blanche Lane, South Mimms, Potters Bar, Hertfordshire, EN~
~ 3QG, United Kingdom.
When the formulation is a liquid formulation, particularly a
spray, the volume of a unit dose typically is in the range 50
to 130 mcl.
~The pH of the composition~ of the present invention can vary
within a broad range according to the chemicophysical
properties of the di~ferent ingredients in the compositions. ~
However, suitably the pH of the compo~ition is in ~he range ~-
from pH 3 to 8, preferably from approximately pH 4 to
~pprsximately pH 7. In order to regula~e the pH and maintain
a suitable value, a buffering agent may be included in the
composition. Examples of buffering agents which may be used
include citrates, for example a mixture of citric acid and
sodium citrate, acetates and phosphates. In addition to a
buffering agent such as those described hereinabove, an
alkali metal hydroxide e.g. sodium hydroxide may be
incorporated to regulate the pH.
The concentration of the benzyl alcohol is between O.l and
5.0% ~w/w) of the total weight of the composition. In a
liquid or gel composition the benzyl alcohol is suitably
present in an amount corresponding to between 0.5 g and 4 g
per lO0 ml of composition. Preferably the benzyl alcohol is
.
W093/06854 ~12 0 7 ~ ~ PCT/EP92/02321
- 13 -
present in an amount corresponding to approximately 2 g/lO0
ml. In suppositories, tablets and soft gelatin capsules for
rectal or vaginal administration the benzyl alcohol is
suitably present in an amount corresponding to between O.l g
s and 1 g per lO0 g of composition. Preferably the benzyl
alcohol is present in an amount corresponding to between O.l
g and 0.5 g per lO0 g.
The concentration of the glycyrrhizinate absorption enhancer
0 typically is at least O.l~ (w/w), suitably O.l to lO~ (w/w),
and preferably 0.2 to 5% ~w~w) of the total weight of the
composition.
Where the composition is a liquid or gel compo~ition, the
glycyrrhizinate suitably is present ~n an amount ~:
corresponding to between 0.5g and 5g per lO0 ml of :~
composition. Preferably the glycyrrhizinate is present in an
am~unt corresponding to approximately 2g/lO0 ml. In
suppositories, table~s and soft gelatin capsules for rectal -
20~ or ~aginal administration the glycyrrhizinate ~ suitably
present in an amount correspondin~ to between O.l g and 2 g
per lO0 g of composition. Preferably the glycyrrhlzinate is
present in an amount corresponding to between 0.2 g and l g
per lO0 g. ~:
:
For aqueous compositions, the final pharmaceutical form, i.e.
liquid soluti~n or gel, can also depend upon the pH, the
ionic strength of the solution and the concentration of -:
glycyrrhizinate. In general, compositions having a pR of
about 5.5 and above will exist as l~quids whllst compositions
having a lower pH value will tend to be more viscous and, at `~
around pH 4.5, will exist in a gel form.
The invention will now be illustrated in greater detail by
the following examples.
W093~068~ 2 ~ 2 0 7 5 5 PCT/EP92/02321
- 14 -
Formulations for na~al, ~ubli~gual, buccal, roctal or
~aginal admini~tration
Rxamples 1-3
s
Table 1
Example No.
1 2 _ 3
Elcatonin ~mcg) 300 300 300 -
(6500 I.U./mg potency)
Ammonium glycyrrhizinate (g) 2 2 2
Citric acid ~mg) 37 37 37
Sodium citrate dihydrate 463 463 463
Sodium chloride (mg) 600 600 600
Benzyl alcohol (g) . 05 1 2
Distilled water q.s. to 100 ml
lN NaOH q.s. to pH 6 ;
The formulations of Examples l to 3 were prepared by mixing
together the ammonium glycyrrhizlnate, citric acid, sodium
citrate dihydrate, sodium chloride, distilled water and
sodium hydroxide in a water bath regulated at a temperature
of about 70C. To the resulting solution cooled to room
temperature, benzyl alcohol and elcatonin were then added.
W093/068~ 2 ~ 2 ~ 7 ~ ) PCTJEP92/0~21
-- 15 --
~xamples 4-l0
The following compositions were prepared according to the
5 method described in Examples l to 3.
Table 2
Example No. ~-
4 5 6 7 8 9 l0
Elcatonin (mcg)3690 3690 3690 7380 7380 7380 7380 ~-
(6S00 I.U./mg -:
potency) :
Ammonium 0.5 2 5 0~5 1 2 5
glycyrrhizinate
(g)
Citric acid (mg) 37 37 37 37 37 37 37
Sodium citrate 463 463 463 463 463 463 463
dihydrate (mg)
_ Sodium chloride600 600 600 600 600 600 600
(mg)
Benzyl alcohol (g)2 2 2 2 2 2 2
Distilled waterq.s. to l00 ml
lN NaOH q.s. to pH 6
W093/068~ 2 1 2 0 7 ~ 5 - 16 - ~CT/EP92/02321
~xamplQQ 11-15
The following compositions were prepared according to the
5 method described in Examples 1 to 3.
Table 3
Example No.
11 12 13 14 15
Elcatonin (mcg) 36903690 73807380 7380
(6500 I.U./mg potency)
Ammonium glycyrrhizinate ~g) 2 2 2 2 2
Citric acid (mg) 37 37 37 37 37
Sodium citrate dihydrate 463 463 463 463 463
(mg)
Sodium chloride ~mg) 600 600 600 600 600
Benzyl alcohol (g) 0.5 4 0.5 1 4
Distilled water q.s. to 100 ml
~lN NaOH q.s. to pH 6
W093J0~ ~ 2 ~ 2 ~PCT/EP92/0~21
- 17 - :
Examples 16-19
Tabl~ 4 . :
-~.
Example No.
16 17 18 l9 :
Elcatonin ~mcg) 3690 3690 7380 7380
~6500 I.U./ms potency)
Ammonium glycyrrhizinate (g) 2 2 2 2
Citric acid tmg) 37 37 37 37 -
Sodium citrate d~hydrate ~mg)463 463 463 463
Sodium chloride ~mg~ 600 600 600 600
Benzyl alcohol (g) 0.5 l 0.5
Methyl p-hydroxybenzoate (mg)130 130 130 130 :
Propyl p-hydroxybenzoate (mg~20 20 20 20
Distilled water q.s. to 100 ml
lN NaOH q.s. to pH 6 ::~
The formulations of Examples 16 to 19 were prepared by mixing
together the ammonium glycyrrhizinate, citric acid, sodiu~
citrate dihydrate, sodium chloride, methyl p-hydroxybenzoate, ~.
0 propyl p-hydroxybenzoate, distilled water and sodium .
hydroxide in a water bath regulated.at a temperature of about
70C. To the resulting solution cooled to room temperature,
benzyl alcohol and elcatonin were then added.
WO 93/06854 2 1 2 ~ 7 5 5 PCI/EP92~0232l
-- 18 --
Example~ 20-21
The following composit~ons were prepared according to the
5 method described in Examples 1 to 3
Table 5
Example No.
21
Elcaton~n (mcg) 3690 7380
(6500 I .U. /mg potency)
Ammonium glycyrrhizinate (g) 2 2
Citric acid (mg) 37 37
Sodium citrate dihydrate (mg) 463 463
Sodium chloride ~mg) 600 600
Benzyl alcohol (g) 2 2
Distilled water q.s. to 100 ml
_ 0.1N NaOH q.s. to pH 4.5
The formulations of Examples 20 and 21 are gels.
W093/06854 5PCT/EP92/02321
Exa~ple 22
Elcatonin (mcg) 7380
(6500 I.U./mg potency)
Ammonium glycyrrhizinate (g~ 2
Acetic acid (mg~ 200
Sodium acetate trihydrate ~mg) 200
Sodium chloride (mg) Ç00 -~
Benzyl alcohol (g) 2 :;
Distilled water q.s. to ml 100 `~
lN NaOH q.s. to pH 5.3
The formulation of Example 22 was prepared by mixing ~ogether
the ammonium glycyrrhizinate, acetic acid, sodium acetate
trihydrate, sodium chloride, disti~led water and sodium ~-
hydroxide in a water bath regulated at a tempera~ure of about ~-
70C. To the resulting solution cooled to room temp~rature,
~ benzyl alcohol and elcatonin were then added~
W093/068~4 2 1 2 0 7 ~ ~ PCT/~P~2~0~321
- 20 -
TRIA~ A
The preparation reported in Example 3 containing ammonium
glycyrrhizinate 2% and benzyl alcohol 2~ as absorption
enhancers, was compared, in a test of pharmacodynamic
activity in rats i.e. lowering of calcium concentration in
the serum, with the following preparations:
a formulation with the same composition as E~ample 3 with the
exception of benzyl alcohol (reference preparation A); a
formulation with the same composition as Example 3 with the
exception of ammonium glycyrrhizinate (reference preparation
B); a formulation with the same composition as Example 3 with
the exception of both benzyl alcohol and ammonium
lS glycyrrhizinate (reference preparation C).
The preparations were administered intranasally with a small
catheter, in the vslume of lO0 mcl/kg body weight
(corresponding to 2 I.U./kg), to groups of lO Sprague Dawley
20 ~ rats weighing 160+20 g. The animals, fasted overnight, were
anaesthetized with tribromoethanol (TBF) 2% (9 ml/kg body
weight~ given i.p.) lS min before receiving elca~onin.
Serum calcium concentration was measured (with an atomic
absorption spectrophotometer ~ARIAN 30/403 on blood samples
obtained in each animal, from the caudal vein, 0, 30, 60, 120
and 180 min after administration of the products.
The results, expressed as residual percentage of serum
calcium concentration as compared with baseline values
(0 time), are reported in Table 6.
In order to evaluate the relati~e efficacy of the
preparations, the AUC ~0-l80 min) values were calculated;
3s being AUC calculated on the residual serum calcium, a lower
AUC is indicative of a greater pharmacodynamic effect. The
AUC (0-180 min) values and the differences (~ AUC) between
the test preparations and reference preparations A and B in ~;
W093/068~ 212 D 7 5 ~ PCT/EP92/02321
- 21 - ;
comparison with that of the reference preparation C, which
does not contain either ammonium glycyrrhizinate or benzyl
alcohol, are reported in Table 7.
,:
The results obtained show a clear effect of synexgism due to
the combination of am~onium glycyrrhizinate and benzyl
alcohol.
o Tabl~ 6
Residual percen~age of serum calcium
as compared.with baseline values ~
0 30 60 120 180 ~::
min min min min
Preparation of this100.0 83.0 73.7 78.7 88.0
invention reported
in Example 3
Reference . 100.0 85.0 76.1 94.7 89.0
preparation A
Reference 100.0 89.6 90.5 92.0 89.5
preparation B
Reference 100.0 101.4 93.5 91.7 91.7
preparation C
W093/068~ PCT/EPg2/02321
21207~5 - 22 -
Table 7
AUC (0-180 min) calculated
on the residual serum
calcium
AUC~ AUC
Preparation of this invention 14,7512,318
reported in ~xample 3
Reference preparation A 15,884lr185
Reference preparation B 16,534535
Reference preparation C 17,069
W093/0~54 2 12 0 7 5 '3 PCT/EP92/02321
- 23 - :
TRIA~ B
The preparation reported in Example 2 containing ammonium
glycyrrhizinate 2% and benzyl alcohol 1% as absorption
enhancers, was compared, in a test of pharmacodynamic
activity in rats i.e. lowering of calcium concentration in
the serum, with the following preparations: :
0 a formulation with the same composition as Example 2 with the
exception of benzyl alcohol (reference preparation A); a
formulation with the same composition as Example 2 with the
exception of ammonium glycyrrhizinate (reference preparation ~:
B); a formulation with the same composition as Example 2 w~th ~
15 the exception of both benzyl alcohol ~nd ammonium .
glycyrrhizinate (reference preparation C). ~:
The testing methodologies were the same described for Trial
A.
The results of residual percentage of serum calcium are
reported in Table 8. The AUC and ~AUC values are reported in ~
Table ~ :
,~;
The results obtained show a clear effect of synergism due to
the combination of ammonium glycyrrhizinate and benzyl
alcohol.
W093/068~ 2 ~ 2 0 7 ~ ~ PcTrEPg2/o232l
- 24 -
Table 8
Residual percentage of serum calcium
as compared with baseline values
0 30 60 120 18~
min min min min
Preparation of this 100.080.2 75.4 77.3 85.0
invention reported
in Example 2
Reference 100.081.4 79.6 89.. 8 86.1
preparation A
Reference 100.087.6 . 92.0 87.6 85.B
preparation B
Reference 100.096.8 87.7 90.4 90.4
preparation C
~able 9
- AUC (0-180 min) calculated on
the residual serum calcium :~
AUC ~A~C
Preparation of this 14,631 1,959 :-
invention reported in
~xample 2
Reference prPparation A 15,535 1,055
Reference preparation B 16,09Q 500
Reference preparation C 16,590
WOg3/068~ 2 12 ~ 7 5 ~PCT/EP92/0~21
- 25 -
Examples 23 and 24
Table 10
:
Example No. -~
23 24
Salmon calcitonin ~mcg) (5S009090 18180 ;`~
I.U.~mg potency)
Ammonium glycyrrhizinate (g) 2 2
Citric acid (mg) 3
Sodium citrate dihydrate (mg)463 463 -
Benzyl alcohol (g~ 2 2
v Sodium chloride (mg~ 600 600
Distilled water q.s. to 100 ml
lN NaOH q.s. to pH 6
The formulations of Examples 23 and 24 are prepared by mixing
~ together the ammonium glycyrrhizinate, citric acid~ sodium
citrate dihydrate, sodium chloride, distilled water and
sodium hydroxide in a water bath regulated at a temperature
of about 70C. To t~e resulting solution cooled to room
temperature, benzyl alcohol and salmon calcitonin are then
added.
WO 93/06854 2 1 2 0 7 5 S PCr/EP92~02321
-- 26 --
`:
Example~ 2~ and 26
Ta~la ll
Example No.
_25_ _ _ 26
Eel calcitonin (mcg) l0000 20000
~5000 I .U. /mg potency)
Ammonium glyGyrrhizinate (g) 2 2
Citric acid ~mg) 37 37
Sodium citrate dihydrate (mg) 4 63 4 63
Benzyl alcohol (g) 2 2
Sodium chloride (mg) 600 600
Distilled water q. s . ~o l00 ml
lN NaOH q. s . to pH 6
The formulations of Examples 25 and 2 6 are prepared by mixing
together the ammonium glycyrrhizinate, citric acid, sodium :~
citrate dihydrate, sodium chloride, distilled water and
sodium hydroxide in a water bath regulated at a temperature
of about 70C. To the resulting solutio~ cooled to room
temperature, benzyl alcohol and eel calcitonin are then
added.
W093/06854 2 12 ~ 7 5 5 PCT/EP92/02321 ~
- 27 -
,~;
Example~ 27 and 28
Table 12
Example No.
27 28
_
Chicken calcitonin II (mcg) 10000 20000
(5000 I.U./mg potency) .
Ammonium glycyrrhizinate ~g) 2 2
Citric acid ~mg) 37 37
Sodium citrate dihydrate ~mg) 463 463
Benzyl alcohol (g) 2 2
Sodium chloride (mg) 600 600
Distilled water q.s. to lQ0 ml
lN NaOH q.s. to pH 6
The formulations of Examples 27 and 28 are prepared by mixing
to~ether the ammonium glycyrrhizinate, citric acid, sodium
citrate dihydrate, sodium chloride, distilled water and
sodium hydroxide in a water bath xegulated at a temperature ;
of about 70C. To the resulting solution cooled to room
temperatur , benzyl alc9hol and chlcken calcitonin II are
then added.
W093/~ 2 12 ~ 7 ~ 5 PCT/EP92/0~21 :
- 28 -
Ex~mple~ 29 and 30
Table 13
Example No.
29 30
Human calcitonin (mg) 250 500
(200 I.U./mg potency)
Ammonium glycyrrhizinate (g) 2 2
Citric acid (mg) 37 37
Sodium citrate dihydrate (mg) 463 463
Benzyl alcohol ~g) 2 2
Sodium chloride (mg) 600 600
Distilled water q.s. to l00 ml
lN NaOH ` q.s. to pH 6 :~
The formulations of Examples 29 and 30 are prepared by mixing
together the ammonium glycyrrhizinate, citric acid, sodium
lO citrate dihydrate, sodium chloride, d.tstilled water and ~:
sodium hydroxide in a water bath regulated at a temperature
of about 70~C. To the resulting solution cooled to room
temperature, benzyl aloohol and human calcitonin are then
added.
W093/068~ 2 1 2 0 7 ~ 5 PCT/EP92/0~2t
- 29 -
Example3 31 and 32
Table 14
Example No.
31 32
Pig calcitonin (mg) (60 I.U./mg 834 1668
poteney)
Ammonium glycyrrhizinate (g) 2 2
Citric acid tmg) 37 37
Sodium citrate dihydrate (mg) 463 463 ~:
Benzyl alcohol (g) 2 2 .
Sodium chloride (mg) 600 600 :
Distilled water q.s. to 100 ml
lN NaOH q.s. to pH 6
The formulations of Examples 31 and 32 are prepared by mixing
~together the ammonium glycyrrhizinate, citric acid, sodium
lo citrate dihydrate, sodium chloride, distilled water and `~
sodium hydroxide in a water bath regulated at a temperature
of about 70C. To the resulting solution cooled to room
temperature, ben~yl alcohol and pig calcitonin are then
added .
W093/06X~ 2 1 2 0 7 5 ~ PCT/EP92/0~21
30 -
PowdQr for na~al admini~tration
Examples 33 and 34
Table 15
Example No.
33 34 ~:~
Elcatonin (mg) (6500 I.U./mg 3.69 7~38
potency)
Ammonium g~ycyrrh~zinate (g) 2.0 2.0
Benzyl alcohol (g) 2.0 2.0
Lactose q.s. ~o (g) 25.0 25.0
~''.
0 The formulations of Examples 33 and 34 are prepared by
wetting the lactose with, benzyl alcohol and with an aqueous
solution of elcatonin and drying under vacuum. The dried -
powder is mixed with ammonium glycyrrhizinate and the final
mixture is placed into hard gelatine capsules t25 mg each
capsule ) .
The powder is administered, fter having pierced the
capsules, using a nasal insufflator.
W093/0~854 2 12 0 7 ~ 5PCT/EP92/02321
- 31 -
Sublingual tablet~
Examples 35 and 36
. .
- Table 16
Example No.
36
Elcatonin ~mg) 7.7 15.4
~6500 I.U./mg potency)
Ammonium glycyrrhizinate (g) 4.0 4.0
Benzyl alcohol (g) 4.0 4.0
Sucrose (g) 35.0 35.0
Mannitol (g) 35.0 35.0
Polyethylene glycol 6000 (g) lO.0 10.0
Lactose q.s. to ~g) 120.0 120.~
lo ~The formulations of Examples 35 and 36 are prepared by mixing
together the sucrose, the mannitol and the lactose. The
resulting mix~ure is wetted with benzyl alcohol and with an
aqueous solution of elcatonin, granulated through a staînless
steel screen and dxied under vacuum. The dried granules are
mixed with polyethylene glycol and ammonium glycyrrhizinate
and then compressed into ~ablets of 120 m~ each.
WOg3/068~ 2 1 2 Q 7 ~ 5 PCT/EP92/02321
- 32 -
Buccal tabletq
~xample-q 37 and 38 ~;
Table 17
': '
Example No.
37 38 ~
Elcatonin (mg) 7~7 15.4 :
(~50~ I.U./mg potency)
Ammonium ~lycyrrhizinate (g) 4.0 4.0
Benzyl alcohol ~g) 4.0 4.0 -
Sucrose (g) 30.0 30.0 :~
Mannitol lg) 35.0 35.0
Polyethylene glycol 6000 (g) 15.0 15.0
Carbopol 934 ~g) 15.0 15.0
Lactose q.s. to (g) 150.0 lS0.0 ;-
0 The formulations of Examples 37 and 38 are prepared by mixing
together the sucrose, the mannitol and the lactose. The ~:
resulting mixture is wetted with benzyl alcoho~ and with an
aqueous solution of elcatonin, granulated through a stainless
steel screen and dried under vaeuum. The dried granules are
mixed with ammonium glycyrrhi7inate, Carbopol and
polyethylene glycol and then compressed into tablets of 150
mg each.
W093/06854 2 I 2 ~ 7 5 ~ PCT/EP92/02321
- 33 -
Oral tablets for colo~ic deli~ery
Exampl9s 39 and 40
Table 18
Example No.
39 40
Elcatonin (mg) 15.4 30.8
(6500 I.U./mg potency)
Ammonium glycyrrhizinate (g) 6.0 6.0
Ben~yl alcohol (g) 4.0 4.0
Pregelatini~ed starch ~g) 80.0 80.0
Magnesium stearate (g) 2.0 2.0
Lactose~.s. to (g) 210.0 210.0
Eudragit S (g) 20.0 20.0
Polyethylene glycol 6000 (g) 2.0 2.0
The formulations of Examples 39 and 40 are prepared by mixin~ .
together the pregelatinized starch and the lactose. The
resulting mixture is wetted with benzyl alcohol and with an
aqueous solution of elcatonin, granulated through a stainless
steel screen and dried under vacuum. The dried granules are
mixed with ammonium glycyrrhizinate and mag~eslum stearate
and then compressed into tablets of 210 mg each.
The tablets are coated with an aqueous suspension of
polyethylene glycol and Eudragit, to a final weight of 232
mg/tablet.
W093/068S4 ~ ?~ ~ 7 ~ ~ PCT/EP92/02321
- 34 -
Dosage form for vaginal administration
ExaMple~ 41 and 42
Table 19
Example No~
41 42
Elcatonin (m~) (6500 I.U./mg15.430.8
potency)
Ammonium gtycyrrhizinate (g~~.0 8.0
Benzyl alcohol (g) 8.0 8.0
Corn starch (g) 180.0 180.0
Adipic acid ~g) 140.0 140.0
Sodium bicarbonate (g) 110.0 110.0
Magnesium stearate (g) 20.0 20~0
Lactose q.s. to ~51 1600.0 1600.0
The formulations ~f ~xamples 41 and 42 are prepared by mixing .
together the ammonium glycyrrhizinate, the corn starch, the
adipic acid and the lactose. The resulting mixture is wetted
with benzyl alcohol and with an aqueous solution of
elcatonin, granulated through a stainless steel screen and
5 dried under ~acuum. The dried granules are mixed with
sodium bicarbonate and magnesium stearate and then compressed
into tablets of 1.6 g each.
2~207~
W093/068~ PCT/EP92/02321
35 -
Do~age form for rectal administration
Example 43
.
Elcatonin (mg)(6500 I.U./mg 15.4
potency)
Ammonium glycyrrhizinate (g) 6.0
Ben~yl alcohol ~g) 4.0
Distilled water (g) 100.0
Hard fat q.s. to ~g) 1500O0
The formulation of Example 43 is prepared by mixin~ toge~her
the ammonium glycyrrihizinate, benzyl alcohol and distilled
water in a water bath regulated at a temperature of about
70C. The solution is cooled to about 40C, elcatonin is
dissolved and then the resulting solution is inco~porated
~ into hard fat melted a~ about 40CO
15 The final mixture is poured into suppository moulds and
cooled to room temperature, thus obtaining suppositories of
1.5 g each.
W093/068~ 2 ~ 2 0 7 ~ .~ PCT/EP92~02321
- 36 -
Formulations for ~aginal or rQctal administration
~xamples 44-46
Table 20
Example No.
44 45 _46 _
Elcatonin (mg) 7.4 ~4.8 29.6
(6500 I.U./mg potency)
Polyethylene glycol 600 (g)550.0 550.0 550.0
Gelucire 44/14 (g) 400.0 40~.0 400.0
Distilled water (g) 42.1 42.1 42.1
Ammonium glycyrrhizinate (g)5.0 5.0 5.0 .:~
Benzyl alcohol (g) 2.0 2.0 2.0
Sodium chloride ~mg) 300.0 300.0 300.0
Sodium citrate dihydrate ~mgl231.5231.5 231.5
Sodium hydroxide ~mg) 350 0 350.0 350.0
- Citric acid (mg) 18.5 18.5 lB.5
lo The formulations of ~xamples 44 to 46 were prepared by mixing
together the ammonium glycyrrhi7inate, citric acid, sodium
citrate dihydrate, sodium chloride, sodium hydroxide,
distilled water and elcatonin in a water bath regulated at a
temperature of about 70C.
The resulting solution, cooled to about 55C, was incorpor-
ated into a mixture of Gelucire, polyethylene glycol and
benzyl alcohol heated to about 55C.
The final mixture, cooled to about 30C, was filled into soft ~:
gelatin capsules ~1 g each capsule).
W093/06854 2 ~ 2 0 7 S S PCT/EP92/0~21
ExamplQs 47-49
~able 21
Example No.
47 48 ~9
. . .
Elcatonin (mg) 7.4 14.8 29.6
(6500 I.U./mg potency)
Polyethylene glycol 600 ~g)400.0 400.0 400.0
Witepsol S55 (g) 450.0 450.0 450.0
Miglyol 812 ~g) 100.0 100.0 100.0
Distilled water (g) 42.1 42.1 42.1
Ammonium .glycyrrhizinate (g)5.0 5.0 5.0
Benzyl alcohol tg) 2.0 2.0 ~.0
Sodium chloride ~mg) 300.0 300.0 300.0 ;-~
Sodium citrate dihydrate 5mg)231.5 231.5 231.5
Sodium hydroxide (mg) 350.0 350.0 350.0
Citric acid (mg) 18.5 18.5 18.5
The formulations of Examples 47 to 49 were prep~red by mixing ~:
together the ammonium glycyrrhizinate, citric acid, sodium
lo citrate dihydrate, odium chloride, sodium hydroxide,
distilled water and elcatonin in a water bath regulated at a
temperature of about 70C.
The resulting solution, cooled to about 55C, was
incorporated into a mixture of Witepsol, Mislyol,
polyethylene glycol and benzyl alcohol heated to about 55C.
;
The final mixture, cooled to about 30C, was filled into soft
gelatin capsules (1 g each capsule~.
W093/068~ ?, l ~, ~ 7 ~ ~i PCT/EP92~02321
- 38 -
Dosage for~ for tran~dermal administration
Example 50
Elcatonin ~mg) 6
(6500 I.U./mg potency)
Ammonium-glycyrrhizinate (g) 2
Benzyl alcohol (g) 2 :
Carbopol 934 (g) 2
Citric acid (mg) 37
Sodium citrate dihydrate (mg) 463
~ Sodium chloride (mg) 600
Distilled water q.s.to 100 g
lN NaOH q.s. to pH 6
., .
:.
The formulation of Example 50 is prepared by mixing together `.
the ammonium glycyrrhizinate, citric acid, sodium citrate
lO ~ dihydrate, sodium chloride, Carbopol 934, sodium hydrQxide
and part of distilled water in a water bath regulated at a
temperature of about 70C. To the res~ltin~ gel, cooled to
room temperature, a solution of elcatonin and benzyl alcohol
in the remaining part of distilled water, is then added.
The final gel is filled into patches of 500 mg each.
W093/ ~ S4 2 1 2 Q 7 ~ .~ PCT/EP92~0232l
- 39 - :
For~ulations ~or nasal, sublingual, buccal, rectal or
~agi~al a~ministration
Exampl~s 51-52
s
Table 22
~ .
Example No.
51 52
Elcatonin ~mcg) (6500 I.U./mg 3690 7380 ~
potency) ~.
Ammonium glycyrrhizinate (g) 2 2
Citric acid (mg) 37 37 `;~
Sodium citrate dihydrate (mg) 463 463
Mannitol~(g) 3.3 3.3
Benzyl alcohol (g) 2 2
Distilled water q.s. to 100 ml
lN NaOH q.s. to pH 6 :
: -~
The formulations of Examples 51 and 52 were prepared by
mixing together the ammonium glycyrrhizinate, citric acid,
sodium citrate dihydrate, mannitol, distilled water and
sodium hydroxide in a water bath regulated at a temperature
of about 70C. To the resulting solution cooled to room
temperature, henzyl alcohol and elcatonin were then added.
W093/0~ ~ PCT/EP92/02321
2~207aS 40 _
Examplas 53-54
Table 23
~
Example No. .
53 _ 54
Elcatonin (mcg) (65QO I.U./mg3690 7380 ~:
potency~
Ammonium glycyrrhizinate (g) 2 2
Citric acid (mg~ 37 37 -
Sodium ci~rate dihydrate (mg) 463 463
Glycine (g) l.~ l.6
Benzyl alcohol (g) 2 2
Distilled water q.s. to lOO ml
lN NaOH q.s. to pH 6
The formulations of Examples 53 and 54 were prepared by
- mixing together the ammonium glycyrrhizinate, citric acid,
o sodium citrate dihydrate, glycine, distilled water and sodium
hydroxide in a water bath regulated at a temperature of about
70C. To the resulting 50lution ccoled to room temperature,
benzyl alcohol and elcatonin were then added.
W093/068~ 2 1 2 0 7 ~ S PCT/EP92/02321
~xamplas 55 - 60
Table 23
Example No.
56 57 58 59 60 :
Elcatonin (mcg)
16500 I.U./mg 1480029600 14800 2~600 14800 29600
potency)
Ammonium
glycyrrhizinate 2 2 2 2 2 2
(g)
Citric acid (mg) 37 37 37 37 37 37
Sodium citrate 463 463 463 463 463 463
dihydrate (mg)
Sod~um chloride 600 600
(mg) :.
mannitol (g) - - 3.3 3.3
glycine ~g) - - - - 1.6 1.6
Benzyl alcohol (g~ 2 2 2 2 2 2
Distilled waterq.s. to 100 ml
lN NaOH q.s. to pH6-
The compositions are prepared in an analogous manner to
Example lo
