Note: Descriptions are shown in the official language in which they were submitted.
7~7
- 2 - HOE 77/F 2~3
The present invention relates to a stable secretin
compositlon, a process for its preparation and its use.
The invention provides a stable secretin composi-
tion for injection purposes, a process for its preparation
and its use as medicament, prophylactic and diagnostic
agent.
:Secretin is a heptacosipeptidamide (Eur,J.Biochem, 15,
513 (19'70)) which is formed in the duodenum and ~lhich,
besides showing other properties, above all stimulates the
hydrogenocarbonate production of the pancreas and inhibits
the action of gastrin, It is therefore appropriate for
the diagnosis and therapy of diseases of the digestive
tralt,
In the above and the following description the term
secretin inclucles the physiologically tolerable salts.
According to the invention there is used natural as ~ell
as synthetic secretin.
It has already been described that secretin is very
unstable and rapidly loses its biological effect especial-
ly in solution at room temperature (Methods in Investiga-
tive and Diagnostic Endocrinology, vol, 2B, page 1067,
North Holland Publishing Co,, Amsterdam, London 1973),
This is why secretin is stored in a lyophilized form and
is dissolved not until shortly before its use.
2~Since secretin is frequently administered in very
small amounts (for example, from 0.025 to 0,2 mg, which
corresponds to about 100 to aoo cu (cu rneaning clinical
unlt)), it is advantageous to add a vehicle substance to
29 the secretin solution prior to lyophili~ation, in ordcr to
, ~
76~7
- 3 - HOE 77/F 263
avoid the rlsk of dust formation.
It has been known from literature that the carrier
substances which are commonly used, such as dextran and
mannitol, adversely affect the stabillty of secretln
(German Offenlegungsschrift No. 2 323 187). According to
the d!ata given in Gastroenterology 57, 767 (1969), cy-
steine hydrochloride is suitable as vehicle with a favor-
able influence on the stability. However, it is a drawback
that the secretin stabilized in this manner cannot be
iodized in a radioactive manner any more, which makes it
unsuitable for use in radioimmuno-assays. Solubility
problems (clouding of the solution) might also arise from
a possible oxidation of the easily soluble cysteine to
give the sparingly soluble cystine.
In German Offenlegungsschrift No. 2 323 187 different
amino acids are tested for their capability of being used
as carrier substances in secretin preparations. In this
connection it was found that at a pM value of from 4 to
4.5 alanine proved to be superior to, for example, glycine,
threonine or valine and that alanine is thus appropriate
as vehicle of stable secretin compositions.
We have now found that the stabillty of secretin com-
positions strongly depends on the pH value and the tempe-
rature of the solution and that glycine buffered to a pH
of 3 is particularly suitable as carrier substance.
The subject of the present invention is therefore
a stable lyophilized secretin composition which has been
adjusted with hydrochloric acid to a pM value of 3 and
29 ~hich optionally contains glycine buffered to pH 3 as car-
.~ . . .
,
~37~7
- 4 - ~OE 7t/F 263
rier substance.
The invention further provides a process for the pre-
paration of a stable lyophilized secretin composition,
which comprises adding secretin to an a~ueous solution ad-
justed with hydrochloric acid to a pH of 3 and cooled toabout 0 to 10C, the solution optionally containing gly-
cine buffered with hydrochloric acid to pH 3, flltering
the solution under sterile conditions, filling it into
bottles or ampules and lyophilizing it.
In Figure 1 the biological activity of a secretin
composition containing 0.1 mg of secretin hydrochloride
and 20 mg of glycine as carrier substance has been illu-
strated. Curves 1 and 2 not only demonstrate the activity
as a function of the pH value of the composition, but also
the dependence on temperature. The composition which has
been lyophilized only after 68 hours o~ standing at room
temperature (curve 2) has lost in activity as compared
with the cornposition which has been lyophilized imrnedia-
tely (curve 1). The maximum activity is found at a pH
value of 3.
In order to avoid the loss in activity due to in-
fluences of temperature, the secretin-containing solution
is lyophilized instantly according to the process of the
invention, if possible.
With a lower secretin dosage (0.025 mg per ampule) it
has been observed that also composltions of a pH value of
3 showed a lower biological effect than was to be expected.
It has been found, however, that these compositions
2g regain their full activity upon adding gelatin partial hy-
.
::
, .
'
~ 7~7
- 5 - HOE 77/F 263
. ..~
drolysate which is cross~linked via diisocyanates (cf. Ger-
man Patents Nos. 1,118,792 and 1,155,134) ((R)Haemaccel),
human albumin or human globin. Evidently the result found
was not due to a deactivation of the secretin, but was
5 caused by an adsorption of the secretin at the glass sur-
face lof the amlpule, which may be prevented andtor undone
by adding proteins or protein-like substances.
For this reason compositions having a lower secretin
content suitably include the above-mentioned additives.
10 The latter are not required in the case of high-dosage
compositions (for example 80o CU and more per ampule).
In order to achieve a protracted action of the secre-
tin, the secretin composition of the invention may be mix-
ed prior to administration with a depot carrier. As depot
15 carrier there is used preferably a mixture of polyphlore-
tin phosphate and gelatin partial hydrolysate cross-linked
with diisocyanates (cf. German Patent No. 2,104,344).
Since the secretin compositions are injected, it is ad-
vantageous to dissolve the lyophilized product prior to
20 its administration in Aqua pro injectione which contains a
physiologically tolerable buffer.
The secretin compositions of the invention are sui-
table as diagnostic agents for the pancreatic functlon and
for the diagnosis of Ulcus duodeni, as therapeutic agents
25 for Ulcus duodeni, Ulcus ventriculi, stress ulcera, bleed-
ings in the gastroduodenal region, reflux oesophagitis,
mucoviscidosis and the Zollinger-Ellison syndrome, as well
as for the prophylaxis of stress ulcera, for example af~er
Zg operations, multiple fractures, burns and septicaemias.
. . , - ,. ,. ; . .
- ..
:: . :~ ..
:
, .
- 6 - HOE 77/F 263
As diagnosti~ agent for the pancreatic function there
is preferably used 2 10 ml ampule with 100 CU. The con-
tents of the ampule are dissolved in 10 ml of physiolog,cal
salt solution and slowly administered intravenously. Also
an infusion for this purpose is possible.
~ he secretin composition of the invention is also
appropriate for the diagnosis of Ulcus duodeni, for fol-
lowing an intracutaneous administration of secretin a
positive skin reaction was observed in Ulcus patients and
! 10 test persons who had a duodena]. ulcer in former years.
This observation is therefore employed for diagnosis.
Secretin is administered in this case in doses of from
preferably 5 to 10 CU.
Bleedings in the gastroduodenal region are treated
with secretin infusions. About 0.5 CU per kilogram of
body weight is administered within one hour. The infusion
period is as a rule 48 hours. For this purpose, the 200
CU ampule is particularly suitable which lasts for about 4
to 5 hours. It is recommended to prepare a fresh solution
every 4 to 5 hours.
In the therapy of Ulcus duodenl and Ulcus ventriculi
there is preferably used a depot carrier according to
German Patent No. 2,104,344 ~hich maintains the secretin
act.ion at the pancreas (hydrogenocarbonate secretion) and
at the stomach (gastrin inhibition) for several hours.
For reasons of stability the depot carrier is not admixed
to the secretin composition, but is only mixed with
secretin immediately ~efore administration. Another rea-
29 son for the separate storage of the depot carrier and the
6~7
KOE 77!F 263secretin composition i3 provided by the opportunity of an
exact secretin dosing which depends on the weight of the
patient. The dose is in this therapy preferably 10 CU
per kilogram of body weight. For this use, for example, a
800 CU ampule is appropriate. The depot carrier and the
secretin composition of the invention are dissolved to-
gether in 1 ml of bidistilled water and are injected sub-
cutaneously. 1 to 2 injections of this depot carrier/se-
cretin mixture are administered daily, and the therapy may
be prolonged up to 6 weeks. This depot composition may
alsol be employed for treating the Zollinger-Ellison syn-
drome, mucoviscidosis and reflux oesophagitis, as well
as for the prophylaxis of stress ulcera, for example after
operations, multiple fractures, burns and septicaernias.
The following Examples serve to illustrate the inven-
tion.
E X A M P L E 1:
.
100 CU ampule
A solution of 20 g of glycine in 400 ml of water which
has been cooled to about 5C is adjusted with 1N HCl to a
pH value of 3. To this solution are added 2.5 g of gela-
tin partial hydrolysate cross-linked with diisocyanates
(Haemaccel(R)) in the form of an about 10 g solution. In
this carrier solution 100,000 CU of secretin hydrochloride
are dissolved. The mixture is then made up with water to
500 ml. The solution is filtered through an appropriate
filter under sterile conditions and is subsequently filled
into ampules of 0.5 ml each and lyophili7ed.
.
~v~ 7
- 8 - P.OE_ 7/F
E X A _ P L ~ 2:
100 CU ampule
The composition is prepared in a manrler analogous to
that of Example 1. Instead of 2.5 g of Haemaccel there is
added 1 g of human albumin.
E X A M P L E 3:
2 CU ampule
The composition is prepared in a manner analogous to
that of Example 1. Instead of 100,000 CU there are added
200,000 CU of secretin llydrochloride.
E X A M P L E 4:
I
200 C_ _ pule
I The composition is prepared in a manner analogou,s to
that of Example 3. Instead of 2.5 g of Haemaccel there is
added of 0.5 g of human globin.
E X A M P L E 5:
800 CU ampule
.
The composition is prepared in a manner analogous to
that of Example 1. Instead of 100,000 CU there are added
80o,000 CU of secretin hydrochloride. The 2.5 g of ~ae-
maccel are not added.
E X A M P L E 6: `
10 CU ampule
. . .
The composition is prepared in a manner analogous to
that of Example 1. Instead of 100,000 CU there are added
10,000 CU of secretin hydrochloride.
E X A M P L E 7:
. _ . ....
Preparation of the depot carrier
29 3 Grams of polyphloretin phosphate are dissol~ed, while
,~
:, , , ~ - :
:.
-
6~7
- 9 - HOR 77/F 263
stirring, in 1N NaOH, and the solution is adjusted to a
pH value of 6.8. To this solution there are added 8 g of
Haemaccel as a ~0 ~ solution. Subsequently the mixture
is made up to 200 ml with distilled water, is filtered un-
der sterile conditions and filled into bottles having arolled edge or 2 ~l each and is lyophilized.
:, :"
~;
