Note: Descriptions are shown in the official language in which they were submitted.
.~09260~ c ase 100-4314
Derivatives of Somatostatin
The present invention relates to new poly-
peptides.
In accordance with the invention there are
provided new compounds of formula I,
:
SB SB
CH2 CH2
Rl-CH-CO-X-Y-Phe-Phe-Trp-Lys-Thr-Phe-Thr-Ser-NH-CH Z
3 14
wherein Rl ls (i) H, ~ .
(li~ NH2 ~ . :
(iii) H-Ala-NH-;
H-Tyr~NH-;
: H-Cys-NH-;
H-Val-NH-;
~ : . H-Lys-NH-;
¦: ~ H-(a-aminO~~iSObUtyryl~NH-;
H-Phe-NH-;
:15~ (iv~ NH2-NH-C0-NH-;
(vj C6H5.NH-CO-Gly-NH-;
H-Ala-Gly-NH-; . ~ :
H-Val-Gly-NH~
: H-a-phenyl-Gly-NH-;
!: . : : ~::
,: ~ '.:
1~9Z~ 100~4314
~vi) R2-A-CO-NH-,
wherein
- R2 is lower alkyl, phenyl, or
- phenyl substituted by halogen,
hydroxy, amino, lower alkyl :
or lower alkoxy, : `
A is a direct bond, alkylene con-
tainin~ up to 6 carbon atoms
or -NH-,
(vii) R2~Ala-NH-; .
R2-A1a-Gly-NH-; ..
wherein R2 is as defined above,
....
(viii) C6H5.A .C0-M-NH-,
whereïn
AI is an alkylene chain of up to
6 carbon atoms, and
-M~ iB -(mono- or di-Hal)-Phe, .
Gly- or -Ala-,
.. . .
~ (ix) ~-(mono- or di-Hal)-Phe-NH-,
: 20 wherein
.
T- is H-, H-Val- or benzoyl-,
., :
:, '
, , ,
,
',' ` ;':,' '
- 2 -
: ' :'
..
',, : ` ' : '
, : . : . :
. .
~ ~ z~ ~ ~ 100-4314
(x) (CH2 ~ CH.A .CO-L-NH,
wherein
AI is as deined above, and
-L- is -Val-, -Val-(mono- or
di-~al)-Phe-, -a-phenyl-Gly-,
and n is an integer between 4 and 10,
each B is hydrogen or the two radicals8form a direct bond,
X is -Lys-, -Nle- or -Cys-, and
~ . Y is -Asn-, -Gln- or -Thr-, and
: 10 - Z signifies the radicals (i) H; (ii) COOH;
(iii) COOR3 (wherein R3 is lower alkyl~;
~iv) -CO-NR4R5 (wherein R4 and R5 independently
are hydrogen or lower alkyl); (v) -CON îCH2)n
(wherein n is as defined above); (vi) CH20H;
(vii) -CO-asparaginol; -CO-leucinol; -CO-iso-
leucinol; -CO-valinol; -CO-norleucinol;
-CO-glutaminol; -CO-threoninol; (vili) -CO-Leu;
. -CO-Serj -CO-Iso-Leu; -CO-Val, and their
.'` corresponding amides, wherein the amide : --
~0 portion signifies the radical -CO NR4R5 or
~CO-N ~CH2)n (wherein n, R4 and R5 are as
defined abov~), or (ix) -CO-ThrOR3 or : .:
-CO~SerOR3 (wherein R3 is as defined above);
_ 3 _ ~ ` :
: . . ,- '
, ~
. ' '
: ' ' "
.~
-
~L~9Z6~JL
100-4314
: '
; (x) -CO-NH-(CH2)k-OH (wherein k is an integer be-
tween 2 and 6), or -CO-NH-CH-[CH2]1 OH
L
[CH2]1 OH
wherein 11 and 12 independently signiy a
whole number fxom 1 to 6);
(xi) -CO-N o;
(xii) -CO-NH-R6 (wherein R6 is a 5- or 6-membered
saturated heterocyclic ring conta.~ning an
oxygen atom as a ring member, or a 5- or 6-
membered lactone ring), wherein in formula I
~ thè unit containing radicals Rl, X, Y and Z,
as well as in the remaining units of the pep-
tlde sequence, at least one unit may be pre-
. ~ .
sent ln D orm, :`
: with~the provisos that when X ls ~ys and Y ls Asn, and all
' ` :~ lS the remainlng units of the peptide se~uence 3 to 14 are
:~ present in the D or L f orm, and when
(i) Rl is H-Ala-Gly-NH- ox
Ala-Gly-NH-, ~
Z is other than COOH, COO~3,
C~2OH or CONR~R5, or, when
ii) Rl is H, NH2 or H-(D)-Ala Gly-NU~
Z iB other than H, COOH, CONR4R5
or (CN~_,CH2)4 or 5 - or, when ~ :
(lii) Rl is H-~-amlno-isobutyryl)-NH- or
: 25 : H~(D)-Ala-NH-,
: Z ls other than H or COOH, or, when
~ 4 _
: ~ -
~92~
100-4314
(iv) Rl is H-Ala-Gly-NH-,
Z is other than H, or when
(v) Rl is H-Ala-NH- or R2-CO-NH-
(wherein R2 is lower alkyl or
unsubstituted phenyl),
Z is other than COOH, and
with the further proviso tha~ when Rl is H-Cys-NH or
H-(D)-Cys-NH and/or X ls -Cys- or -(D)-Cys, B-B is other
than a direct bond.
10In the compounds of the formula I, when X is
Lys and Y is Asn, Z is pref~ra~ly qelected from the
radicals deined under (viij, (viii), (ix) and (x~ above.
~ Halogen signifies bromine, fluorlne, and
preferably chlorine. The significance of lower alkyl or
lower alkoxy comprises up to 6 carbon atoms, but prefer-
ably signlfies 2 or 1 carbon atom.
When ~2 is substituted phenyl, this is pre-
ferably mono- or disubstit~ted, especially, however
monosubstituted. A substituent is preferably present in
~20 the para position.
: I :
A when alkylene or A preferably contains
up to 3 carbon atoms. In regard to the significance ~
viii3, A preferably signifies trimethylene. In regard
to the significance (x3, A preferably signifles methylene.
Examples of -(mono- or di-Hal)-Phe-moieties
are -(2Cl)-Phe-; -(3C13-Phe-; -(4Cl)-Phe- or
(3,4-di-Cl)-Phe-.
. :~: - : : .
~ - 5 -
. . ~ .
~260~ 100-~314
n preferably signifies an integer 5 or 4.
Rl preferably signifies, e.g.,
H-(mono- or di-Hal)-Phe-NH-, phenyl-CO-NH-,
H-Val (mono- or di-Hal)-Phe-NH-, phenylbutyryl-NH-
or a radical containing -(D)-Ala, benzoyl-(mono- or
di-Hal)-Phe-NH-, cyclohexylacetyl-a-phenyl-Gly-NH-,
cyclohexylacetyl-Val-(mono- or di-Hal)-Phe-NH~,
cyclohexylacetyl-Val-NH-, H-Val-NH, H, H-(a-amino-
isobutyryl)-NH-, H-(mono- or di-Hal)-Phe-NH-.
X preferably signifies Lys and especially Nle.
Y preferably signifies Gln and especially Asn.
~ preferably signifies 2 ox 3
When Z is significance tx) or (xii), this
preferably contains a moiety -CO-NH-CH-CH2-0-
and signifies, for example, -CO.NH.CH(CH2.0H)CH2.CH2.CH2.0H;
-CO-NH-CH(CH20H).CH2.CH2.OH; -CO-NH-CH.CH2.CH2.CO.OjCH2;
-CO-NH.CH CH2.CO.O.CH2; -CO-NH.CH.CH2.CH2ØCH2;
-CO-NH.fH CH2.CH2.CH2 O.CH2.
Z preferably signifies other than H,
...
CH20H, COOH or COOR3.
~ '
In the preferred significance indicated above
- ~ the aminoacids may be present~ in the D instead of the
L form.
: : ' .'
- 6 -
: :
:, ~
: : '
1' . .. . " ., . . ;. . ,. , , . , .. . . , . . , . . ~., . ~ ... .. .
~Z6~ 100-~314
The present invention comprises processes for
the production of compounds of the above formula. They
may be produced in accordance with known methods for the
. synthesis of compounds of this type or obvious chemical
-~ 5 equivalents thereof.
The polypeptides or derivatives thereof of the
above formula may be produced, for example, by a process
comprising
a) removing at least one protective group from a protected
peptide having the sequence indicated in formula I, ox
. b3 link~ng together by an amide bond two peptide units,
; each of which contains at least one amlnoacid and
which is ln protected or unprotected form, the peptide
, unlts being such that the aminoacid sequence given in
formula I is obtained, and then if necessary carrying
. out process step a), or . .
c~ converting one group Z ~f an unprotected or protectea
peptide of formula I in to another group Z havlng the
definitlon indicated above, and if necessar~ carr~ing
out process step a), or ;
~ d) oxidizing a peptide of formula I whexeln B is hydrogen,
to produce a peptide of formula I wherein B-B is a : `
direct bond. ~ :
.
~ _ 7 _
'` ` ` `
- .:
` .
:
. ~ .. .. ... .. . : . .. .. : , : . . . . . : : :.: . :.. .
- ~9260~ 100-431~
The above are methods known in peptide
chemistry; they may be carried out in a manner analogous
to the processes described in the following Examples. :
Insofar as the production of the starting
materials is not particularly described, ~hese compounds
are known or may be produced and purified in accordance
with known methods. These compounds may also be produced
in a manner analogous to the processes described in the
Examples.
Thecompounds may exist in salt form or in the
form of complexes thereof.
The salts of hydrochloric acid and acetic acld
may particularly be mentioned as acid addition salts.
Complexes are understood to be the complex-like
compounds of known type, but not of clear structure, which
are formed from compounds of formula I upon adding
certain inorganic or organic substances. Such inorganic
substances are compounds which are derived from metals,
such as calcium, magnesium, aluminium, cobalt and
~.
-~ 20 particularly zinc, especially difficultly soluble salts
~', such as phosphates, pyrophosphates and polyphosphates,
as well as hydroxides of these metals, in addition alkali
metal polyphosphates. Organic substances are, for example,
'9` non-antigenic gelat~nes, e.g. oxypolygelatine, polyvinyl-
pyrrolidone and carboxymethyl cellulose, in addition
,
~ 7 :
~' ~ ' ' ' .
', , ' '
- ',' ,:
" ' .
~ ~ Z~ 0~ 100-~31~
sulphonic acid or phosphoric acld esters of alginic acid,
dextrane, polyphenols and polyalcohols, especially
polyphloretine phosphate and phytinic acid, as well as
polymerization products of aminoacids, e.g. protamine,
polyglutaminic acid or polyasparaginic acid.
In the following non-limitative Examples all
temperatures are indicated in degrees centigrade and
are uncorrected.
The following abbreviations are used:
Cbo = carbobenzyloxy
MBzl = ~-methoxybenzyl
Iabu = H-(~-amino-isobutyryl)-
(4Cl)Phe - ~-chlorophenylalanyl
AcOH = acetic acid
OCP = trichlorophenyl ester
OSu = O~succinimide
~' '' :'-
All the final compounds are characterized in
hydrochloride salt form. The aminoacids are present in
the ~ form unless otherwise indicated.
i
. ..
.
'','-',.~
,~ '
`, ' ' ~'~ :''
_ g ~
' ' ",
' . :
, . , ~
' ~ . '
~9Z6~ 100-4314 .-
;
In the following Table ~a] D0 refers to
AcOH 1~ v/v (c = 1) unless otherwise indicated.
.
(1) refers to AcOH 20% v/v (c = 1)
(2) refers to AcOH 1% v/v (c = 0.25)
(3) refers to AcOH 30% v/v (c = 1~
~4) refers to AcOH 40~ v/v (c = 1)
(5) refers to AcOH 50% v/v (c = 1)
- ~ ~
~,
'.
.. '' .
.
': " '
'~', .
-: .
10 - .:
i,~ : ' ~ '
:
i : :
.. . .
26~ 00~43l4
EXaMPLE 1:
a~O~
480 mg of Cbo-(4Cl)Phe -Cy5 (MB zl )-Lys(BOC)-Asn-
Phe-Phe-Trp-Lys(Cbo)-Thr-Phe-Thr Ser-Cys(MBzl)-OBzl, 410
mg of H-Try-OH, 320 mg of H-Cys-OH.HCl and 1,2 cc of thio-
anisole are dissolved at 0 in 5 cc of trifluoroacetic
acid, and 10 cc of 2 molar boron tris(trifluoroacetate) ~-
in trifluoroacetic acid are added. The mixture is shaken
for one hour at room temperature and cooled. 50 cc of
tert~butyl alcohol are added, concentration is efEected
by evaporation, in a vacuum and a solution of 0.01 molar
2~mercaptoethanol in ether is added to the residue.
Centrifuging is subsequently effected. The solid residue
ls dlssolved ~n a small amount of 10~ acetic acid and is
:, . .
purified by chromatography on "Sephadex G" 25 in a sys-
tem of 0.01 molar 2-mercaptoethanol in 10~ acetic acid.
The residue may also be dissolved in 30% acetic acid or
in~a mixture of n-butanoljacetic acid/water (e.g. 4
and purlfied by chromatography on "Sephadex G" 25 using
0.01 M 2-mercaptoethanol in 30% acetic acid or in the
mixture o~ n-butanoljacetlc acid/water. The fractions con-
; taining the desired product are combined, dissvlved in
::
0.1 normal hydrochloric acid and lyophilized, whereby tlletitle compound is obtained.
M.Pt, 195 (decom~.); [~12 = -20.5 in AcO~ (c = 1).
The starting material ls produced as follows:
-- 11 --
Trademark for a dry, unsoluble, powdered material derlved
from dextran; lt is used in chromatography.
,
, - , . . . . ~: ,.. . .
26e~L
100~4314
, ' :
a) Cbo-Lys(BOC)-Asn-Phe-OMe
________________________
74.8 g of Cbo-Lys(BOC)-ONP are reacted in
a solution of 51 g of H-Asn-Phe-OMe . HCl and 21 cc of
triethylamine in 300 cc of dimethyl formamide. After
working up, the title compound is obtained.
M.P. 175; [1 D= ~4 4 in dimethyl formamide (c = 1.0).
b) H-Lys(EOC)-Asn-Phe-OMe
_______________
~ 2 g of Cbo-Lys(BOC)-Asn-Phe-OMe are sus-
pended in a solution of 2 liters of dioxane and 400 cc
of water. Palladium charcoal is added and hydrogenation
is effected at normal pressure. After working up, the
title compound is obtained.
' M.P. 122; [] D- -8.3 in dimethyl formamide (c = 1.0).
:,
c) Cbo-(4Cl)Phe-C~s (MB21) -~H-NH
____________ 2
20 g of H-Cys(MBzl)-OMe.CH3S03H and 7 cc
of triethylamine are disso:Lved in 120 c~ of dimethyl
formamide, and 18.2 g of Cbo-(4Cl)Phe-ONP are added.
.
The mixture is allowed to stand for 16 hours and is
concentrated at room temperature, whereby Cbo-(4Cl)Phe-
Cys(MBzl)-OMe (M.P. 115) is obtained after worXing up.
, -
The latter is dissolved in methanol and hydrazinehydrate is added. The mixture is allowed to stand at
room temperature for one day. The title compound is
. ~ ~
': : , ,
. : :
- ~ 12 -
'~
~ .
~ ~. . ., ~ .
~IL0~2~1
100~431
obtained after working up.
M.P. 158; [a~ OD= -9.9 in dimethyl formamide (c = 1).
d) Cbo-(4Cl)Phe-Cys(MB~l)-Lys(BOC)-Asn-Phe-NH-NH
____________~______-_-_ 2
6 g of Cbo-(4Cl)Phe-Cys(MBzl)-~H-NH2 are
dissolved in 80 cc of dimethyl formamide, the solution
is cooled to -20, 6 cc of 5.3 normal hydrochloric acid
in dioxane are added and then 1.3 cc of tert.butyl
nitxite are added and stirring is effected at -20 for
- 10 minutes. After the addition of 5.5 cc of triethyl-
amine at -20, 8.4 g of H-Lys(BOC)-Asn-Phe-OMe are
added, the mixture is allowed to stand over night at 0, ..
concentration is effected, and water is added at 0
up to pH 2. The precipitate is filtered off and washed
with water. The resulting Cbo-(4Cl)Phe-Cys(MBzl)-
1 15 Lys(Boc)-~sn-Phe-OMe is recrystallized from methanol,
:` is then dissolved in dimethyl formamide,and 7 cc of
hydrazine hydrate are adaea. The mixture is allowed
to stand at room temperature for one day, is con-
centrated, and the residue is recrystalliæed from ~ .
methanol, whereby the title compound is obtained.
M.P. 260; [a] D- -23~ in dimethyl formamide (c = 1.0).
e) BOC-Lys(Cbo)-Thr-Phe-Thr-Ser-Cys(MBzl)-oBzl
_____ _______________--_-------- .. ::
2.3 g of BOC-Lys(Cbo)-Thr-Phe-Thr-Ser-NH-
NH2 are reacted with 1.86 g of H-Cys(MBzl)-OBzl in a : -.
''.. " ':
:
; 13 - :
: .
I
' : ~: ' '
, .
~ . , , , . ,.,.... .. ; ., . . , . . . ,, .,. - ..
- 11 09Z6iD1
100-~314
manner analogous to that described in section d), where-
by the title compound is obtained.
M.P. L69~j [~¦ D- -16 in dimethyl formamide (c = 1).
f) BOC-Trp-Lys(Cbo)-Thr-Phe-Thr-Ser-Cys~MBzl)-OBzl
___________________._________________ ________._
2.3 g of BOC-Lys(Cbo)-Thr-Phe-Thr-Ser-
Cys(MBzl)-OBzl are dissolved in a mixture of 10 cc of
methylene chloride and 6 cc of trifluoroacetic acid
and the solution is allowed to stand at room temperature
for 25 minutes. H-Lys(Cbo)-Thr-Phe-Thr-Ser-Cys(MBzl)-
OBzl ~ trifluoroacetate is subsequently precipitated
with ether, filtered off and washed out well with
ether. The residue is dissolved in 7 cc of dimethyl
formamide, l.5 g of BOC-Trp-OCP and 0.3 cc of
triethylamine are added and the mixture is allowed to
stand at room temperature over night. The product is
precipitated with ether/ethyl acetate (l : l) and is
filtered. Drying is effected, whereby the title com-
pound is obtained.
,~ M.P. 167; [~ D- -17.5 in dimethyl formamide (c = l).
, 20 g) BOC-Phe-Tr~-Lys(Cbo)-Thr-Phe-Thr-Ser-Cys(MBzl)-oB
~ __ __ _~__ __________._ _________~_ __ _ _ ____ _ _
36 g of BOC-Trp-~s(Cbo)-Thr-Phe-Thr-Ser-
Cys(MBzl)-OBzl are reacted with 20 g of BOC-Phe-ONP
~ in a manner analogous to that described in section f);
,.~
. . .
' :` : '' .
- 14 -
,
1 ~`
.
2~
100-~314
the title compound is obtained.
M.P. 210; [~] D- 15 in dimethyl formamide (c = 1.0).
h) Cbo-(4Cl)Phe-Cys(MBzl)-~ys(Boc)-Asn-phe-phe-Tr
~ys(Cbo)-Thr-Phe-Thr-Ser-Cys(MBzl)-OBzl
~: ----_ _ _ _ _ _ _ _ _ _
1.5 g of BOC-Phe-Trp-Lys(Cbo)-Thr-Phe-
Thr-Ser-Cys(MBzl)-OBzl are dissolved in 30 cc of
methylene chloride and 30 cc of trifluoroacetic acid.
The mixture is allowed to stand at room temperature
for 25 minu-tes, is concentrated by evaporation in a
vacuum and precipitated with ether. After filtration,
washing with ether and drying, H-Phe-Trp-Lys(Cbo)-Thr-
Phe-Thr-Ser-Cys(MBzl)-OBzl . trifluoroacetate is ob-
tained.
0~9 g of Cbo-(4Cl)Phe-Cys(MBzl)-Lys(BOC)-
Asn-Phe-NH-NH2 are dissolved in 20 cc of dimethyl
formamide, the solution is cooled to -20~, 0.6 cc of
5.3 normal hydrochloric acid in dioxane are added and
then 0.1 cc of tert.~utyl nitrite is added and the
mixture is stirred at -20 for 15 minutes. 0.7 cc of
triethylamine and 1.5 g of H-Phe-Trp-~ys(Cbo)-Thr-Phe-
Thr-Ser-Cys(MBzl)-OBzl trifluoroacetate are added.
The mixture is allowed to stand at 0 over night, is
concenkrated by evaporation in a vacuum, is stirred -
:
with 100 cc of water, filtration is effected, the resi-
. .
~ .
. .
, ~ - 15 -
,
._ . :
.
. .. . . . . ~ . .
~ Z61)~
100-4314
,
due is washed with water and then wi-th methanol, heating
: in methanol and filtration are effected, whereby the
title compound is obtained.
M.P. 270; [a~ D- -19.3 in dimethyl formamide (c = 1).
',
The following compoundSo~
formula A are obtained in analogous manner after building
up the corresponding protected compounds and splitting
off the protective group,
.~' .
P-Phe-Phe-Trp-Lys-Thr-Phe-Thr-Ser-Q A
. ~ .
~ ~ .
, ~ .
: . . .
', . , ::
'~; , : ~
:
:1 :
~:
~ 16 - ~ `
' ~ :` , '
-~ ~Z~9Z~ZO~
100-~l31~
No. _ _ _ l~}~O
Al ~ NH-CO-Cys-Lys Asn .Cys-OH -17 2/
A2 ~ -NH-CO-Cys Lys-Gln Cys-OH 150 ~
~3 II-Tyr-Cys-Lys-Asn Cys-OH 160 /
. . -20.0
A4 NH2 ~ CO~Cys-Lys-Asn Cys-OH -19.Z~O
A5 ~ (CH2)3-CO-Cys-Lys-Asn Cys-O~I -33 1
A6 ~ (CH2)3-CO-Gly-Cys-Lys-Asn Cys-OH -27 7
' . A7 ~ NH-CO-Gly-Cys-Lys-Asn Cys-OH -34 3
A8 H-Cys-Cys-Lys-~sn Cys-N~2 -24 5
~ A9 H-Cys-Cys~Asn -Cys-NH2 -23 3
: ~ A10 H-Lys-Cys-Lys-Asn C~s-NH2 ~27.8
All ~I-(4Cl)Phe-Cys-Nle-Asn ~ Cys-NH2 170
A12 N-(4Cl)Phe-Cys-Lys-Asn Cys-NH -23 2
Z A13 H-Iabu-Cys-Lys-Asn : Cys-NH2 -21.0
A14 H-Iabu-Cys-Lys-Thr Cys-NH 165 /
~, 20.~
:`, ' : ' .
- ~
:', ~ :
260~
100-431~l .
Ex __ l~J2(/
~; ~ A15 ~ -(CH2)3-co-~cl)phe-cys~Lys-Asn Cys-NH2 -1~ 7/
: A16 ~ -(cH2)3-co-Gly-cys-Nle-Gln Cys-NH -1~.7 :~
Al ~ (CH2)3-C0-Cys-Lys Asn Cys NK _19 r~o
~ Al ~ NH-C0-Cys-Lys~Asn Cys-NH2 -25 3/~1)
: A19 ~ NH-C0-Cys-Lys-Gln Cys--NH -29.5 - :
A2 ~ NH-CO~Cys-Nle-Asn Cys-NH~ -10.7
A21 ~ NH-C0-Gly-Cys-Nle-Gln Cys-NH2 1134/ ~:
, A2 ¦~H2-N~-C0-Cys Lys-Asn Cys-NH 130 /
: A2 ~ (CH2)3-CO-(D)Ala-Cys-Lys~Asn Cys-NH -13.7
~:~ A24 8enzoyl-Cys-Nle-Asn Cys-NH lGO /
. A25 8enzoyl-~y5-Lys-Asn Cys-NH2 -37~9
A26 ~ C~-(4Cl)Phe-Cys-Nle-Asn Cys-NH
¦A27~h-~D)-Al -Gly-C s-Lys-Asn ¦ Cys-Val-NH2 ¦24270~(/2) ¦
A2 H-Ala-Gl~-Cys-Lys-Asn Cys-N ~ 210 /
,~ . -45(2)
, j
~ - 18 ~
.. . .
, .
.: , .
,,_ ,r~ ~ ' '
~9Z6(~ 100-4 31~
_ ~
Ex . _ Q [c~]20
_ ___ __ _~___ ~
A29 1~-(4Cl)Phe~Cys-Nle^Asn Cys-Asparagi- 155 / --
; llol -22.5
A30 ~ abu-Cvs-Nle~Asn Cys~Asparayi- 150 /
nol -44.3
A31 Il--Iabu-Cys-Lys--Asn Cys-Asparagi- 160 /
nol- -~7.4
A32 H-(D)-Ala-Gly-Cys-Lys-Asn Cys-Aspara~i- 150 /
. nol -52.7 ~:-
A33 H^(D)-~la~Gly Cys-Nle~Asn Cys-Asparagi- 165 / ~:
; . nol _49.50(1)
A34 ~I-(D)~Val-Cys~Lys-Asn Cys-NH 175 /
. 47.0~
A35 ~ Phenylc3~ycyl)-Cys-Lys-Asn Cys-NH2 ~33 3
A36 H-(D)~Val-(4Cl)Phe-Cys Lys-Asn Cys-NH 165 /
~37 ~I-(D~-Ala-Cys-Lys-Asn Cys-NH ~30.9
i A3~ H~Cys~Lys-Asn Cys-NH 180 /
;l A39 Cyclohexy].acetyl-(D~-Val-Cys-Lys-Asn Cys-NH2 ~19.80(3)
A40 Cyc].ohexylacetyl~ -Phenylglycyl)-Cys- Cys-NH 1~5 /
Lys Asn -19.~(5
A41 Cyclohexylacetyl~(D)-Val-(4Cl)Phe-Cys- Cys-NH 165 /
Lys-Asn 2 -23.70~3)
A42 8~nzoyl-(3,~-dichlo~^o~Phe-Cys-I.ys-P.sn Cys-NE~2 -22 5/(~) -:.
: A~ 3 Renzoyl- ( 3, 4 -dichloro) Phe-Cys-Nle-~sn Cys-NH -2~.So(4)
,~ . , .~ A44 H-(D)-Ala-Gly-Cys-Nle-Asn Cys~NH ~23.90(4)
, ~ ,
`1 ~ ' '''.,.. '
,''' ~ ' , .
~' ~ ' `' .
z~
100-431
: No, . ~ ~ I )2
~ . _ _ ._
~45 H-(D)-Phe-Cys-Nle-Asn Cys-NH -29 1(3
A46 H-Phe~Cys-Nle-Asn Cys-NH2 1730~/(3
. . ~47 H-(3Cl)Phe-Cys~Nle-Asn Cys Asparagi- 165 /
: . nol _35,0(3
A~ H-(2Cl)Phe-Cys-Nle~Asn Cys-~sparagi- 170 /
. nol -29.8(3
A49 H-(3,~diCl)Phe-Cys-Nle-Asn Cys-Asparagi- 162 /
. nol CH2-l=0 -25,5(3 :.
~ASO 1 - ~ 'O ~Cys- I l Cl~
. .
, ' , - '
. ~ , , .
.
~ ' ' . . . ' .
'`' . , "
, .
.
., ' :
-
.'`~, . . .. .
- :, ,.
~ ~ .'''' '.
- - 20 - .
,, , .::
, '
'
26~ loo-43l4
EXP~MPLE 2:
-.
r - - _
H-(D)-Ala-Gly-Cys-Nle-Asn-Phe-Phe-Trp-Lys-Thr-Phe-Thr-Ser-Cys- - :
~ asparaginol
; 0.4 g of the compound of Example A33 are
dissolved in a mixture of methanol and 0.5& acetic acid,
and the pH is adlusted to 6 by the addition of an~onium
hydroxide solution with stirring. A 0.1 molar potassium
ferricyanide solution is sLowly added with stirring until
the yellow colouration romains. After stir*ing for a fur- ;
ther 15 minutes, the pH is adjusted to 4-5 with acetic
acid. Filtration is effected through a short column of
"BLo Rad AG" 3/x4 (Cl~ form) [- slightly basic anion ex-
change resin]. The filtrate ls then allowed to flow
slowly at 0 through a column with "Bio Rex 70"
, ~ .
(-COOH form) [- sllghtly acid cation exchange resin].
The column is then eluted with an acetic acid gradient
with increasing concentrations of acetic acid or with an
, :
acetic acid/pyridine grad~ent. The fractions containing
the desired product are combined and lyophilLzed.
The lyophiIization product is dissolved ln a smalL amount
of n-butanol/glacial acetic acld/water (2 : 1 : l) or
` 30~ acetic acid and purified by chromatography on
~; "Sephadex"G 25 in the
., :
.`,~ `` : :: :
:
: - 21 ~
* Trademark
** Trademark
' .
' ,
~2~ 100-~31~
same mixtu.re. The frac-tions containing the desired
product are combined and lyophilIzed, whereby the
title compound is obtained.
[a] D- ~36 (c = l in 95% acetic acid).
The corresponding compounds having an
S-S- bridge are produced in analogous manner from the
corresponding compounds having a free SH function
(E~amples l, A 1-', A 10-32, A 34-50).
'
,
. , -' '.
. ` ' '
'
''` ~ '`':
` - 22 - '
',:
~: .
,; ' ` '. ~ .
, ~ :
:
0~
100-4314
The compounds of formula I exhibit pharmacologi-
cal activity. In particular they inhibit growth hormone
secretion, as indicated in standard tests.
Male rats anaesthetized with "Nembutal" are
administered with the peptide by injection in the jugular
vein. The rats are decapitated 15 minu~es after admini-
stration, collecting the blood. The growth hormone concen-
tration in the blood serum is determined in conventional
manner by radio immunoassay.
The compounds are therefore indicated for use as
agents for the inhibition of growth hormone secretion and
possibly as agents ~or the treatment of Diabetes ~lellitus,
Acromegaly and Angiopathy.
For this use an indicated daily dose is from
, 15 about 0.07 to about 70 mg, conveniently admlnistered in
`~ divided doses 2 to 4 times a day in unlt dosage form con-
talning from about 0.02 to about 35 mg, or in sustained
release form.
~he compounds of A29, A47 and A48 exhibit
èspecially interesting activity.
The compounds of formula I may be administered
in pharmaceutically accept~ble acid addition salt or com-
plex~ form. Such acid addition salt or complex form exhibit ~-
the same order of activity as the free base forms and are
readily prepared in conventional manner.
, :
23 ~
; ~ * Irademark for pento~arbital, either ln the calcium or
-~ sodium salt form; it is a hypnotic. l ~
. ~ ,.
~: : .
0~
100-431~
The present invention al50 provides A pharmaceutical
composition comprising a compound of formula I, in free
base form, in complex form, or in pharmaceutically
acceptable acid addltion salt form, in association with
a pharmaceutical carrier or diluent. Such compositions
may be in the form of, for example, a tablet or a solution
for injection or infusion, buffered e.g. at from pH 5 to
8.
The present invention also comprises pharma-
ceutical preparations containing compounds of formula I,
wherein Rl is H~(a-amino)-isobutyryl-NH- or H-(D)-Ala-N~I,
X is-Lys; Y is~Asn7 Z is -CONR4R5 or -CO-N (CH2~4 or 5
Each B preferably signifies hydrogen.
,
:~ -
' . ' '"
~..,
- .
~: .
-;:
- 24 - ~
. . .
.
.
