Peptide derivatives

The present invention provides a sugar derivative of a biologically active peptide, which derivative has a prolonged duration of action when compared to the non-sugar modified peptide, and contains at least on one of the amino acid units a sugar residue which is attached to an amino group thereof by a coupling other than a direct N-glycosidic bond, and additionally, when it is a condensation product of a carboxyl group containing sugar and a peptide with less than 8 amino acid units, by a coupling other than a direct amide bond.

The present invention relates to peptides, their production, pharmaceutical 
preparations containing them, and their use as medicaments. 
The present invention provides a sugar derivative of a biologically active 
peptide, which derivative has a prolonged duration of action when Compared 
to the non-sugar modified peptide and contains at least on one of the 
amino acid units a sugar residue which is attached to an amino group 
thereof by a coupling other than a direct N-glycosidic bond, and 
additionally, when it is a condensation product of a carboxyl group 
containing sugar and a peptide with less than 8 amino acid units, by a 
coupling other than a direct amide bond. 
Hereinafter these compounds are referred to as compounds of the invention. 
By non-sugar modified peptide is meant the structurally corresponding 
peptide not having the sugar residue or residues. Hereinafter this is 
referred to as the unmodified peptide. 
We have moreover found that the compounds of the invention show 
particularly interesting and surprising pharmacological properties, 
especially a longer duration of action, e.g. as described hereinafter. 
We have found that incorporation of a sugar residue or residues, even when 
bound in a different manner to normal glycosylation e.g. of Asn or Ser, 
induces these properties. 
It is preferred to introduce these sugar residues on amino groups of amino 
acids remote from the active site of the peptide. 
The term peptides as used herein include: peptides (e.g. di-, 
tri-peptides), oligo-peptides, polypeptides, and proteins. Preferably the 
peptide is of more than 7 amino acid units. Conveniently the peptide is of 
8 to 32 amino acid units. The term amino acid unit as used herein also 
includes an amino alcohol unit, e.g. a reduced amino acid. 
The term biologically active peptides is used herein to cover in particular 
compounds having pharmacological or therapeutical activity, e.g. compounds 
which have hormonal, enzymatic or immunomodulatory activity, or which 
stimulate or inhibit such activity. These biologically active peptides 
encompass natural peptides isolated from nature or fermentation of cells, 
e.g. produced through genetic engineering, or synthesized and also their 
derivatives or analogues. 
By derivatives and analogues is understood in particular natural peptides, 
wherein one or more amino acid units have been omitted and/or replaced by 
one or more other amino acid radical(s) and/or wherein one or more 
functional groups have been replaced by one or more other functional 
groups and/or wherein one or more groups have been replaced by one or 
several other isosteric groups. In general, the term covers all modified 
derivatives of a biologically active peptides, which exhibit a 
qualitatively similar effect to that of the unmodified peptide. 
The sugars used may be e.g. any known mono- or oligosaccharide, especially 
a mono-, di- or triose or a derivative thereof, e.g. an amino- and/or 
carboxylic acid and/or reduced and/or esterified derivative thereof. 
The sugar may be coupled e.g. to a N-terminal amino group and/or to at 
least one amino group of the peptide present in the side chain thereof. 
The sugar may be coupled by one of its functional groups to the peptide 
either directly or indirectly by a bridging member e.g. an alkylene 
carbonyl group. 
This coupling may be made in conventional manner, especially as hereinafter 
described. 
In a preferred group of the compounds of the invention the sugar residue is 
attached to an amino group of the peptide by a coupling other than a 
direct N-glycosidic or direct amide bond. 
A group of the compounds of the invention are preparable by an Amadori or 
Heyns rearrangement. 
The invention also provides oral pharmaceutical preparations containing a 
compound of the invention especially those having at least 8 amino acids 
units. 
The present invention provides in particular the following sugar 
derivatives of biologically active peptides of formula I 
a) 
##STR1## 
wherein 
##STR2## 
is the deoxy residue of a ketose, the residue being linked via the 
CH.sub.2 group to the NH group of a biologically active peptide, and 
P is the residue of a biologically active peptide of formula NH.sub.2 --P, 
wherein the NH group is located on the N-terminal end or in a side chain 
of the peptide P, 
b) of formula II 
##STR3## 
wherein 
##STR4## 
is the deoxy residue of an aldose, the radical being linked via the free 
bond to the NH group of a biologically active peptide, and 
P is the residue of a biologically active peptide of formula NH.sub.2 --P, 
wherein the NH group is located on the N-terminal end or in a side chain 
of the peptide P, 
c) of formula III 
EQU G.sub.3 --CO--NR.sub.y --P III 
wherein 
G.sub.3 CO is the residue of a uronic acid, or of a polyhydroxymono- or 
di-carboxylic acid, 
R.sub.y is hydrogen, alkyl with 1 to 3 C-atoms or alkanol with 1 to 4 
C-atoms, and 
P is the residue of a biologically active peptide of formula NH.sub.2 --P, 
containing at least 8 amino acid units 
wherein NR.sub.y is located at the N-terminal end or in a side chain of the 
peptide P 
d) of formula IVa, IVb, IVc or IVd 
##STR5## 
wherein P denotes the residue of a biologically active peptide of formula 
H.sub.2 N--P, 
##STR6## 
are sugar residues. R.sub.y is hydrogen, alkyl with 1 to 3 C-atoms or 
alkanoyl with 1 to 4 C-atoms, and Q, Q', Q" and Q'" are groups coupling 
the peptide residue with the sugar residue, 
wherein the NH group bonded to P is located at the N-terminal end or in a 
side chain of the peptide, or 
e) of formula Va or Vb 
##STR7## 
wherein Y is H.sub.2 or H, OH 
c is 2 or 3 or 4 
P is a residue of a biologically active peptide of formula H.sub.2 N--P 
wherein the NH group bonded to P is located at the N-terminal end or in a 
side chain of the peptide P, and any one of the free hydroxy groups in the 
polyol moiety of the compounds of formula V is optionally bound in 
glycosylic manner to a reducing mono-, di- or oligosaccharide or amino 
sugar, 
as well as the acid addition salts and complexes of these polypeptide 
derivatives, with the provisos that 
a) in the above-mentioned compounds of formula I, P is other than a radical 
of a gastrin peptide with a C-terminal group ending in -Asp-Phe-NH.sub.2, 
b) when in the compounds of formula IVb Q' is a phenyl-ring containing 
divalent radical or when in the compounds of formula IVd Q'" is the 
residue of an aliphatic dicarboxylic acid, then P--NH.sub.2 is not a 
natural insulin, 
c) when in the compounds of formula III G.sub.3 --CO is the residue of an 
optionally N-acylated muramic acid then the second amino acid residue at 
the N-terminal end of the peptide P--NHR.sub.y should not be the residue 
of an amino dicarboxylic acid. 
A gastrin is a peptide which increases gastric acid secretion. 
All the above-mentioned sugar residues may be mono-, di- or 
oligosaccharides. These sugars may contain heptoses, hexoses and/or 
pentoses, which may exist as pyranoses or furanoses. 
In all the formulae I to V mentioned above only one sugar moiety per 
peptide residue has been shown. However, the invention also covers sugar 
derivatives of peptides having more than one free amino groups on the 
peptide residue, these derivatives containing e.g. 2,or 3 sugar residues 
per peptide residue. 
The invention provides additionally all biologically active peptides which 
have more than one sugar residue which are linked as defined above. 
The sugar polypeptides preferably contain 1 to 3 mono-saccharide residues, 
which may be joined together as a disaccharide or tri-saccharide. 
In all the above-mentioned compounds, the line .about. means that the bond 
may be in the .alpha.- or .beta.-position. 
In formula I 
##STR8## 
preferably is a) a residue of formula 
##STR9## 
wherein one of radicals G.sub.a and G.sub.b is hydrogen and the other is 
OH, 
one of radicals G.sub.c and G.sub.d is hydrogen and the other is OH or 
O-glycosyl, wherein the glycosyl radical is derivable from a reducing 
mono-, di- or oligosaccharide, 
one of radicals G.sub.e and G.sub.f is hydrogen and the other is OH, 
one of radicals G.sub.g and G.sub.h is hydrogen and the other is hydrogen 
or CH.sub.2 OH 
e.g. wherein radicals G.sub.a to G.sub.h are selected such that the residue 
of formula VIa corresponds to a radical which is obtainable by means of an 
Amadori rearrangement from a natural or a synthetically accessible mono-, 
di- or oligosaccharide. 
The following residues may be mentioned as examples of sugar residues of 
formula VIa: 
Deoxyfructosyl, deoxytagatosyl, deoxysorbosyl, 
.alpha.-glucosyl-(1-4)-deoxyfructosyl, 
.alpha.-glucosyl(1-4)-.alpha.-glucosyl(1-4)-deoxyfructosyl. 
b) a residue of formula VIb 
##STR10## 
wherein one of radicals G.sub.a and G.sub.b is hydrogen and the other is 
OH 
one of radicals G.sub.c and G.sub.d is hydrogen and the other is OH or 
O-glycosyl, wherein the glycosyl radical is derivable from a reducing 
mono-, di- or oligosaccharide, 
one of radicals G.sub.e and G.sub.f is hydrogen and the other is hydrogen, 
COOH, CH.sub.2 OH, CH.sub.2 --O--P(O)--(OH).sub.2 or CH.sub.2 O-glycosyl, 
wherein the glycosyl radical is derivable from a reducing mono-, di- or 
oligosaccharide, 
e.g. wherein radicals G.sub.a to G.sub.f are selected such that the radical 
of formula VIb corresponds to a radical which is obtainable by means of an 
Amadori rearrangement from a natural or a synthetically accessible mono-, 
di- or oligosaccharide. 
Residues of formula VIb may be obtained for example by means of an Amadori 
rearrangement from saccharides such as gentiobiose, melibiose, ribose, 
xylose or from uronic acids such as glucuronic acid or galacturonic acid. 
In formula II, 
##STR11## 
preferably is a) a residue of formula VIIa 
##STR12## 
wherein one of radicals G.sub.a or G.sub.b is hydrogen and the other is a 
free bond, 
one of radicals G.sub.c or G.sub.d is hydrogen and the other is OH, 
one of radicals G.sub.e or G.sub.f is hydrogen and the other is OH or 
O-glycosyl, wherein the glycosyl radical is derivable from a reducing 
mono-, di- or oligosaccharide, 
one of radicals G.sub.g and G.sub.h is hydrogen and the other is CH.sub.2 
OH, or CH.sub.2 ---O-glycosyl, wherein the glycosyl radical is derivable 
from a reducing mono-, di- or oligosaccharide, e.g. 
wherein radicals G.sub.a to G.sub.h are selected such that the radical of 
formula VIIa corresponds to a radical which is obtainable by means of a 
Heyns rearrangement from a natural or a synthetically accessible mono-, 
di- or oligoketose. 
b) a residue of formula VIIb 
##STR13## 
wherein one of radicals G.sub.a and G.sub.b is hydrogen and the other is a 
free bond, 
one of radicals G.sub.c and G.sub.d is hydrogen and the other is OH, 
one of radicals G.sub.e and G.sub.f is hydrogen and the other is CH.sub.2 
OH or CH.sub.2 --O-glycosyl, wherein the glycosyl radical is free bond, 
derivable from a reducing mono-, di- or oligosaccharide 
e.g. wherein radicals G.sub.a to G.sub.f are selected such that the radical 
VIIb corresponds to a radical which is obtainable by means of a Heyns 
rearrangement from a natural or a synthetically accessible mono-, di- or 
oligoketose. 
Residues of formula VIIa or VIIb may be obtained for example by means of 
Heyns rearrangement from sugar such as D-fructose, lactulose, L-sorbose, 
D-tagatose or D-ribulose. 
In formula III, the polyhydroxymono- or -dicarboxylic acid, e.g contains at 
least 3 hydroxy groups and may also contain further substituents, e.g. 
amino or acetylamino groups. 
Examples of such polyhydroxycarboxylic acids are: 
the "onic acids" derived from sugar, such as gluconic acid, or "aric acids" 
such as glucaric acid, furthermore, quinic acid, acetylmuranic acid, 
acetylneuraminic acid or D-glucosaminic acid. 
Examples of uronic acids are glucuronic and galacturonic acids. 
In the compounds of formula IV, G.sub.4, G.sub.4 ', G.sub.4 " and G.sub.4 
'" have the definitions given above for G.sub.1, or G.sub.2. 
The radical Q or Q' joins a NH.sub.2 group of the peptide with a NH.sub.2 
or HO group of the sugar residue, and is e.g. the radical of a 
dicarboxylic acid or preferably a --C.sub.b H.sub.2 --CO-radical wherein b 
is 0 to 6. The radical may be branched. Q' denotes for example a 
##STR14## 
or in particular a --C.sub.b H.sub.2b --CO-- radical (b e.g. 1 to 6). Q is 
e.g. a --CH.sub.2 --CO-moiety. Especially Q is --CO-- or --CS--. 
--NH--Q"-- and --NH--Q'" denote radicals which join a NH.sub.2 group of 
the peptide with the sugar residue, especially radicals of 
w-aminocarboxylic acids. They may signify for example a --NH--C.sub.b 
H.sub.2b --CO-- radical. 
In the case of a compound of formula V, preferred are those of formula Va, 
especially wherein n is 3. 
All the above-mentioned residues P are residues of biologically active 
peptides. Such peptides include all natural and synthetic peptides (also 
derivatives and analogues thereof (see the beginning of this description) 
having hormonal, enzymatic or immunomodulatory activity. This activity may 
be both stimulating and inhibiting. The following may be mentioned as 
examples of such peptides: somatostatin, calcitonin, oxytocin, 
vasopressin, insulin, LH, LHRH, GRF, gastrin, substance P, cathepsin, 
encephalins, as well as all derivatives and analogues of these peptides 
which have similar activity to the peptides or having antagonising 
activity. 
The compounds of the invention preferably contain at least 8 amino acid 
units, e.g. 8 to 32, especially 8 to 20, in particular 8 to 10 amino 
acids. 
Preferred peptides are those of formula I and II. 
In the above and following formulae, for the sake of simplicity, the sugar 
radical is usually only represented by the structure of pyranose. 
Naturally, the furanose and open chain structures are also included in the 
invention, provided that they exist for the relevant sugars. 
The present invention includes processes for the production of compounds of 
the above formulae. They may be produced by methods which are generally 
known for the synthesis of compounds of this kind. 
The compounds of the invention may be produced for example as follows: 
a) at least one protecting group, which is present in a sugar derivatized 
peptide, is removed, or 
b) two peptide units, each of which contains at least one amino acid or an 
amino alcohol in protected or unprotected form and one peptide unit 
contains the sugar radical, are linked together by an amide bond, wherein 
the peptide bond is in such a way that the desired amino acid sequence is 
obtained, and stage a) of the process is then optionally effected, or 
c) at least one optionally protected residue is introduced into a protected 
or unprotected peptide and stage a) of the process is then optionally 
effected, or 
d) a functional group of an unprotected or a protected sugar derivatized 
peptide is converted into another functional group or removed, so that an 
unprotected or a protected glycosylated peptide is obtained, and in the 
latter case stage a) of the process is effected, or 
e) a sugar derivatized peptide, in which the mercapto groups of Cys 
radicals exist in free form, is oxidised to produce a peptide in which two 
Cys radicals are joined by a S--S-- bridge. 
As mentioned in the beginning of this description the term "sugar" as used 
herein also covers sugar derivatives such as amino sugars, oxidized and 
reduced sugars or esterified sugars. 
The above reactions may be effected in conventional manner analogously to 
the processes described in the following examples, in particular process 
a) and b) may be effected according to the synthesis of the invention 
described hereinafter. Where desired, in these reactions, protecting 
groups which are suitable for use in peptides or sugars may be used for 
functional groups which do not participate in the reaction. The term 
protecting group includes a polymer resin having functional groups. 
The compounds of formula I may be produced by reacting a protected peptide 
having a free amino group in a slightly acidic medium with a reducing 
mono-, di- or oligosaccharide or a corresponding uronic acid or ester 
thereof (Amadori rearrangement), and subsequently removing the protecting 
groups. 
This reaction may take place in a conventional manner for the Amadori 
rearrangement. The acid added may be e.g. glacial acetic acid. When 
reacting with uronic acid, an additional acid can be dispensed with. It is 
preferable to use an excess of carbohydrate, e.g. ten equivalents for one 
equivalent of peptide compound. The reaction may be carried out in a polar 
solvent such as methanol, preferably at temperatures of ca. 60.degree. to 
70.degree. C. 
The compounds of formula II may be produced by reacting a protected peptide 
having a free amino group in a slightly acidic medium with a ketose (Heyns 
rearrangement). The reaction can be carried out under the same conditions 
as for the Amadori rearrangement (see above). 
The compounds of formula III can be produced by reacting a protected 
peptide having a free amino group with an acid of formula G.sub.3 --COOH 
or a reactive derivative of such an acid, and then removing the protecting 
group(s). This may be a conventional amidation reaction, which can be 
effected in known manner. The acid halides can be used in particular as 
the reactive derivatives of carboxylic acids. The amides can e.g. also be 
produced with the free acids in the presence of hydroxybenzotriazole and 
dicyclohexylcarbodiimide. 
The compounds of formula IVa, IVb, IVc and IVd may be produced by 
a) reacting the peptide first of all with the bridge member and then 
reacting the product with the sugar, or 
b) reacting the sugar first of all with the bridge member and then reacting 
the glycolysed bridge member with the peptide. 
These reactions may be effected in conventional manner. Generally the 
amide, ester or acetal compounds of the invention are major products. The 
compounds of the invention may be purified in conventional manner. 
Compounds of formula IVa wherein Q is --CO-- or --CS-- may be produced for 
example by coupling the corresponding glycosylisocyanate or 
glycosylisothiocyanate of formula 
##STR15## 
wherein L is O or S and G.sub.4 is as defined above and wherein the free 
hydroxyl groups present in G.sub.4 are protected, e.g. by acylation, 
to a peptide P--NH.sub.2 in protected form, and thereafter splitting off 
the protecting groups. 
This reaction may be effected in conventional manner for the production of 
urea derivatives. 
Compounds of formula IVc and IVd may be obtained by means e.g. of an 
Amadori or Heyns rearrangement, e.g. as described above for the production 
of compounds of formula I and II. 
A compound of formula Va or Vb may be produced e.g. by 
a') reductive amination of an aldose, deoxyaldose or ketose with the 
peptide P--NH.sub.2, or 
b') reducing the hemi-acetal group in a compound of formula I or II, 
wherein if desired-any reactant may be temporarily protected. 
The reductive amination and reduction may be effected in conventional 
manner. The reductive amination may be effected for example with 
NaBH.sub.3 CN. The preferred pH is 7. The reduction of the hemi-acetal 
group may be effected with borohydrides, for example with NaBH.sub.4. The 
preferred pH is about 6. 
Insofar as the production of the starting materials is not particularly 
described herein, it is known or may be produced in conventional manner, 
e.g. using methods known in the literature or described herein for 
analogous compounds, or by the synthesis of the invention described 
hereinafter. 
One preferred class of compounds of the invention comprises the sugar 
derivatives of somatostatin peptides, e.g. of 4 to 9 amino acids. The term 
somatostatin peptides includes its analogues or derivatives thereof. 
Especially preferred are the sugar derivatives of compound of formula 
VIII: 
##STR16## 
wherein A is hydrogen, alkyl with 1 to 3 C-atoms or alkanoyl with 1 to 4 
C-atoms, 
&gt;N--CH(Z.sub.1)--CO is 
1) a (L)- or (D)-phenylalanine residue which is optionally substituted by 
halogen, NO.sub.2, NH.sub.2, OH alkyl with 1 to 3 C-atoms and/or alkoxy 
with 1 to 3 C-atoms, or 
2) the residue of a natural lipophilic .alpha.-amino acid or of a 
corresponding (D)-amino acid, other than that given under 1), 
wherein 
Z.sub.1 in &gt;N--CH(Z.sub.1)--CO-- represents the residue of an amino acid 
residue defined under 1) and 2), 
A' is hydrogen or alkyl with 1 to 3 C-atoms, 
Y.sub.1 and Y.sub.2, independently of one another, are 
1) hydrogen 
2) 
##STR17## 
wherein m is a whole number from 1 to 4, 
R.sub.a is CH.sub.3 or C.sub.2 H.sub.5 and 
R.sub.b is H, CH.sub.3 or C.sub.2 H.sub.5, or 
3) 
##STR18## 
wherein n is a whole number from 1 to 5, or 4) --CO--NHR.sub.c wherein 
R.sub.c is a straight-chain or branched alkyl radical with 1 to 6 C-atoms, 
or 
5) 
##STR19## 
wherein R.sub.d is the residue of a natural .alpha.-amino acid (incl. 
hydrogen) which is located on the .alpha.-C-atom, and R.sub.e is an alkyl 
radical with 1 to 5 C-atoms, 
6) 
##STR20## 
wherein R.sub.a ' and R.sub.b ', independently of one another, are 
hydrogen, CH.sub.3 or C.sub.2 H.sub.5, 
R.sub.8 and R.sub.9, independently of one another, are hydrogen, F, Cl, Br, 
alkyl with 1 to 3 C-atoms or alkoxy with 1 to 3 C-atoms, 
p is 0 or 1, 
q is 0 or 1, and 
r is 0, 1 or 2, 
or Y.sub.1 and Y.sub.2 together denote a bond, 
B is Phe or Phe which is substituted in the phenyl radical by F, Cl, Br, 
NO.sub.2, NH.sub.2, OH, alkyl with 1 to 3 C-atoms or alkoxy with 1 to 3 
C-atoms, 
C is L- or D-Trp which is optionally substituted in the benzene ring by F, 
Cl, Br, NO.sub.2, NH.sub.2, OH, alkyl with 1 to 3 C-atoms or alkoxy with 1 
to 3 C-atoms, 
D is Lys, wherein the .alpha.-amino group may be substituted by methyl, 
E is Thr, Ser, Val, 
F is COOR.sub.1, CH.sub.2 OR.sub.2, CO--NR.sub.3 R.sub.4 or 
##STR21## 
R.sub.1 is hydrogen or alkyl with 1 to 3 C-atoms, R.sub.2 is hydrogen or 
the radical of a physiologically acceptable, physiologically hydrolysable 
ester, 
R.sub.3 is hydrogen, alkyl with 1 to 3 C-atoms, phenyl or phenylalkyl with 
7 to 10 C-atoms, but when R.sub.4 denotes --CH(R.sub.5)--X.sub.1, it only 
denotes hydrogen or methyl, 
R.sub.4 is hydrogen, alkyl with 1 to 3 C-atoms or 
##STR22## 
R.sub.5 is the residue of a natural amino acid (including hydrogen) which 
is located on the .alpha.-C-atom, or a HO--CH.sub.2 --CH.sub.2 -- or 
HO(--CH.sub.2).sub.3 radical, wherein the group IX may have the L- or 
D-configuration, 
X.sub.1 is COOR.sub.1, CH.sub.2 OR.sub.2 or 
##STR23## 
R.sub.6 is hydrogen or alkyl with 1 to 3 C-atoms, R.sub.7 is hydrogen, 
alkyl with 1 to 3 C-atoms, phenyl or phenyl alkyl with 7 to 10 C-atoms, 
wherein the radicals B, D and E exist in the L-form and the radicals in 
positions 2 and 7, as well as the radicals Y.sub.1 4) and Y.sub.2 4) exist 
independently in the D- or L-form, as well as salts and complexes of these 
compounds. 
Such compounds are disclosed in U.S. Pat. No. 4,395,403 the contents of 
which including the examples thereof are incorporated herein by reference. 
In the polypeptide derivatives of the above formula VIII, the following 
definitions or combinations thereof are preferred: 
If &gt;N--CH(Z.sub.1)--CO-- has definition 1), this residue preferably is a 
(L)- or (D)-phenylalanine or a (L)- or (D)-tyrosine residue (wherein Z, 
signifies benzyl or p--OH), especially the (D)-pbenylalanine residue. 
If &gt;N--CH(Z.sub.1)--CO-- has definition 2), the residues in which Z.sub.1 
is alkyl with 3, preferably 4, or more C-atoms, e.g. up to 7 C-atoms are 
preferred. 
The &gt;N--CH(Z.sub.1)--CO-- radical most preferably is a residue defined 
under 1). 
Y.sub.1 and Y.sub.2 preferably have the significances given above under 1, 
2 or 4. Especially they form a bond together. 
B preferably denotes Phe or Tyr 
C preferably denotes -(D)-Trp 
D preferably denotes Lys 
E preferably denotes Thr 
F preferably denotes 
##STR24## 
especially 
##STR25## 
wherein the residue --CH(R.sub.5)--X.sub.1 preferably has the 
L-configuration 
R.sub.3 preferably denotes hydrogen, 
R.sub.5 denotes CH.sub.2 OH, 
##STR26## 
i-butyl, CH.sub.2 CH.sub.2 OH or (CH.sub.2).sub.3 --OH, especially 
CH.sub.2 OH or 
##STR27## 
in particular 
##STR28## 
X.sub.1 preferably denotes 
##STR29## 
or CH.sub.2 OR.sub.2, especially CH.sub.2 OR.sub.2, R.sub.2 preferably 
denotes hydrogen. 
R.sub.2 as the residue of an ester preferably denotes HCO, alkyl-carbonyl 
with 2 to 12 C-atoms, phenylalkylcarbonyl with to 12 C-atoms or benzoyl. 
The residues in positions 2 and 7 preferably have the L-configuration. 
Especially preferred sugar somatostatin derivatives are those which have a 
sugar residue on the N-terminal amino group, e.g. compounds of formula 
##STR30## 
Especially preferred are compounds of formula VIIIa, VIIIb, VIIIe and 
VIIIf. 
A group of compounds comprises those of formula VIIIpa 
##STR31## 
wherein A.sup.p is the deoxy radical of a ketose or a corresponding uronic 
acid, the group being linked by a CH.sub.2 group to the NH group, 
said desoxy group being obtainable by an Amadori reaction of an aldose or a 
corresponding uronic acid with the free amino group of the somatostatin, 
and 
Z.sub.1, A', Y.sub.1, B, C, D, E, Y.sub.2 are as defined above with respect 
to formula VIII. 
Another group of compounds comprises compounds of formula VIIIPb 
##STR32## 
wherein G is an acyl radical of an uronic acid, a 
polyhydroxycyclohexanecarboxylic acid, N-acetyl muraminic acid or 
N-acetyl-neuraminic acid, 
A is hydrogen, alkyl with 1 to 3 C-atoms, or alkanoyl with 1 to 4 C-atoms, 
Z, A', Y.sub.1, B, C, D, E. Y.sub.2 and F are as defined above. 
Conveniently G is glucuronic acid, galacturonic acid or quinic acid. 
Another group of compounds comprises those of formula VIIIpc 
##STR33## 
wherein Q is hydrogen or the glycosyl group of a mono-, di- or 
oligosaccharide, 
n is a whole number from 1 to 6, 
A is hydrogen, alkyl with 1 to 3 C-atoms or alkanoyl with 1 to 4 C-atoms, 
Z, A', Y.sub.1, B, C, D, E, Y.sub.2 and F are as defined above. 
A further preferred class of compounds of the invention comprises the sugar 
derivatives of calcitonins. 
The term calcitonin embraces calcitonins which are naturally occurring 
(whether extracted from natural sources, cell cultures etc or produced 
synthetically) and derivatives and analogues. 
The natural calcitonins which may be used include, human, salmon, eel, 
chicken, beef, sheep, rat or porcine calcitonin, especially human, salmon, 
chicken and eel calcitonins. 
Derivatives and analogues of these calcitonins include in particular 
natural calcitonin structures, wherein one or more amino acid radicals are 
replaced by one or more other amino acid radicals and/or the S--S-- bridge 
is replaced by an alkylene bridge, and/or wherein one or several amino 
acid radicals have been omitted. 
Especially preferred are the sugar derivatives of calcitonins of the 
following formula X 
##STR34## 
wherein R is H or R"CO 
R"CO is the acyl radical of a carboxylic acid 
Y.sub.1 ' is the radical located on the .alpha.-C-atom of a .alpha.-amino 
acid, 
Y.sub.2 ' the radical located on the .alpha.-C-atom of a .alpha.-amino 
acid, 
##STR35## 
--CH.sub.2 --S--S--CH.sub.2 --CH.sub.2 --COOH, --(CH.sub.2).sub.s --COOH 
or --CH.sub.2 --S--Y.sub.3, 
Y.sub.3 is alkyl with 1 to 4 C-atoms; benzyl which is optionally 
substituted by methyl or methoxy; or CH.sub.3 CONH--CH.sub.2 --, 
o is a whole number from 1 to 4 
A.sub.6 is Thr or D-Thr 
s is a whole number from 3 to 5 
A.sub.8 is the aminoacyl radical of a neutral, lipophilic L-.alpha.-amino 
acid 
A.sub.9 is the aminoacyl radical of a neutral, lipophilic L- or 
D-.alpha.-amino acid, and 
Z.sub.1 is a polypeptide radical which is located in positions 10 to 31 of 
a natural calcitonin or a derivative or analogue thereof, which has 
hypocalcemic activity, 
wherein the 1 to 4 Y.sub.1 ' radicals in formula X, independently of one 
another, may be the same or different and, with the exception of the 
aminoacyl radical A.sub.8, all amino acid radicals in formula X may have 
the L- or D-configuration, as well as salts and complexes of these 
compounds. 
Such compounds are described for example in GB 2,184,729 A the contents of 
which as well as the specific examples are incorporated herein by 
reference. 
Z.sub.1 in formula X signifies a peptide radical which may be present in 
positions 10 up to 31 in various known calcitonins, e.g. in human, salmon, 
eel, chicken, beef, sheep, rat or porcine calcitonin, as well as in 
derivatives and analogues of these calcitonins, having similar activity. 
By derivatives and analogues of these calcitonins are understood 
especially natural calcitonins, wherein one or more amino acid radicals 
are replaced by one or more other amino acid radicals, or the S--S-- 
bridge is replaced by an alkylene bridge, or wherein one or more amino 
acid radicals have been omitted. These peptide radicals Z.sub.1 normally 
comprise 22 amino acids, but they may also contain a correspondingly 
smaller amount of amino acid radicals by omitting one or several amino 
acid radicals (des-aminoacyl derivatives), 
Z.sub.1 preferably denotes 
a) 
Gly-Thr-Tyr-Thr-Gln-Asp-Phe-Asn-Lys-Phe-His-Thr-Phe-Pro-Gln-Thr-Ala-Ile-Gl 
y-Val-Gly-Ala 
b) 
Gly-Lys-Leu-Ser-Gln-Glu-Leu-His-Lys-Leu-Gln-Thr-Tyr-Pro-Arg-Thr-Asp-Val-Gl 
y-Ala-Gly-Thr 
c) 
Gly-Lys-Leu-Ser-Gln-Glu-Leu-His-Lys-Leu-Gln-Thr-Tyr-Pro-Arg-Thr-Asn-Thr-Gl 
y-Ser-Gly-Thr 
Compounds of formula X wherein Z.sub.1 has the definition given under b) or 
c), preferably those wherein Z.sub.1 has definition c) are especially 
preferred. 
R"CO is preferably the acyl residue of an aliphatic, cycloaliphatic, 
aromatic or heterocyclic carboxylic acid. 
R" is preferably 
a') saturated or unsaturated, straight-chain or branched alkyl with 1 to 17 
C-atoms, especially saturated alkyl with 3 to 9 C-atoms, 
b') cycloalkyl with 5 to 7 C-atoms or cycloalkylalkyl wherein the 
cycloalkyl group contains 5 to 7 C-atoms and the alkyl radical contains 1 
or 2 C-atoms, 
c') adamantyl, adamantylmethyl or adamantylethyl, or 
d') phenyl, benzyl or phenethyl. 
In the above-mentioned definitions for R", the alkyl, cycloalkyl or phenyl 
radicals may be substituted by the usual substituents, e.g. by halogen, 
NO.sub.2, OH, alkoxy, etc. 
The residue R"CO may be e.g. the .alpha.-desamino residue of a natural 
.alpha.-amino acid. For R", definitions a'), b') and c') are preferred. 
Y.sub.1 ' and Y.sub.2 ' as radicals which are found on the .alpha.-C-atom 
of an .alpha.-amino acid are in particular the radicals which are bonded 
to the .alpha.-C-atom of a natural .alpha.-amino acid, but radicals of 
other .alpha.-amino acids may also be contemplated, e.g. of 
3-cyclohexylalanine or of an .alpha.-aminoisobutyric acid. 
When o in formula X signifies 4, 
a) the N-terminal aminoacyl radical (corresponding to the second amino acid 
radical in the sequence of the natural calcitonins) is preferably Ser, Gly 
or Ala, 
b) the second aminoacyl radical (corresponding to the third amino acid 
radical in the sequence of the natural calcitonins) is preferably Asn or 
Ser, 
c) the third aminoacyl radical (corresponding to the fourth amino acid 
radical in the sequence of the natural calcitonins) is preferably Leu, 
Asn, Ser, Phe, D-Leu or the radical of cyclohexylalanine, 
d) the fourth aminoacyl radical (corresponding to the fifth amino acid 
radical in the sequence of the natural calcitonins) is preferably Ser or 
Ala. 
When o in formula X is 3, the N-terminal, the second and the third amino 
acid radicals have the same preferred definitions as above for the case 
when o=4 under b). 
When o in formula X is 2, the N-terminal and the second amino acid radicals 
have the same preferred definitions as above for the case when o=4 under 
c) and d). 
When o in formula X is 1, the N-terminal and the second amino acid radical 
is preferably Ser or Ala. 
A.sub.6 is preferably Thr 
##STR36## 
preferably denotes Cys, a derivative of cystein as given above for Y.sub.2 
', or a neutral lipophilic .alpha.-amino-acyl radical, especially, Ala or 
another neutral lipophilic .alpha.-aminoacyl radical, in particular Ala 
A.sub.8 is preferably the aminoacyl radical of a neutral lipophilic 
.alpha.-amino acid, especially Val or Gly 
A.sub.9 is also preferably the aminoacyl radical of a neutral lipophilic 
.alpha.-amino acid, especially Leu or Phe 
In the compounds of formula X, o is preferably 2, wherein R signifies H or 
R"CO, or in particular, o is 1 and R is R"CO. 
All the amino acid radicals preferably have L-configuration. 
The glycosylated calcitonins (including derivatives and analogues) are 
especially those which are glycosylated on the N-terminal amino group or 
on one or several amino group(s) in one or several side chain(s), e.g. 
compounds of formulae 
##STR37## 
Calc denotes the residue of a natural calcitonin or of a derivative or 
analogue of such a calcitonin, which is bonded to the sugar residue via an 
amino group on the N-terminal end or in a side chain. 
The calcitonin derivative of formula X may be produced by methods which are 
generally known for the synthesis of polypeptides of this kind. The 
polypeptides of the above formula may be produced for example as follows: 
a) at least one protecting group, which is present in a protected 
polypeptide in the sequence given in formula X, is removed, or 
b) two peptide units, each of which contains at least one amino acid or 
derivative thereof, as described for formula X in protected or unprotected 
form, are linked together by an amide bond, wherein the peptide bond 
should be made in such a way that the amino acid sequence contained in 
formula X is obtained, and stage a) of the process is then optionally 
effected, or 
c) a compound of formula X, wherein R denotes hydrogen, in protected or 
unprotected form, is reacted with an acid of formula R"COOH or with a 
reactive derivative of such an acid, and stage a) of the process is 
optionally effected, or 
d) in order to produce a compound of formula X wherein Y.sub.2 ' denotes 
##STR38## 
or CH.sub.2 --S--S--CH.sub.2 --CH.sub.2 --COOH, either a compound of 
formula XII 
##STR39## 
in protected or unprotected form is reacted with a compound of formula 
XIII 
##STR40## 
wherein R.sub.10 is a group which facilitates the formation of a S--S-- 
bridge with the S-atom of the other CH.sub.2 SH group in the peptide of 
formula XII, 
R.sub.11 signifies hydrogen or an amino-protecting group 
R.sub.12 signifies OH or a protecting group for the carboxyl group, and 
V signifies hydrogen or a NH group, or 
a compound of formula XIV 
##STR41## 
in protected or unprotected form, wherein R.sub.10 is defined as above, is 
reacted with a compound of formula XV 
##STR42## 
and then stage a) of the process is optionally effected. 
If the production of the starting products is not specifically described, 
these compounds are known or may be prepared and purified by the usual 
methods. The end products of formula X can similarly be purified in the 
usual way, so that they contain less than 5% polypeptide by-products. The 
peptides used as starting products for processes a) and b) can similarly 
be produced in known manner in solution or by the solid phase process. 
Production of peptide units which contain a --CH.sub.2 --S--S--CH.sub.2 
--CH.sub.2 --COOH or CH.sub.2 --S--S--CH.sub.2 --CH(NH.sub.2)--COOH group 
as the Y.sub.2 ' radical, may take place analogously to the 
above-mentioned process d). 
In this process d), compounds of formula XIII or XIV are used, in which 
R.sub.10 denotes the known radicals which react with mercaptans whilst 
forming a S--S-- bond. R.sub.10 is in particular S-alkyl, --S--COOalkyl, 
##STR43## 
or S--SO.sub.3 --. 
In these radicals, alkyl is especially lower alkyl with 1 to 4 C-atoms. The 
introduction of these radicals into compounds having free SH groups may be 
effected analogously to methods which are known in sulphur chemistry. 
A further preferred class of compounds comprises a group of LH RH 
antagonists. 
Preferably the compounds include sugar derivatives of compounds of formula 
XVI 
EQU R.sub.1 -A.sub.1 -B.sub.1 -C.sub.1 -D.sub.1 -E.sub.1 -F.sub.1 -G.sub.1 
-H.sub.1 -I.sub.1 -K.sub.1 -NH.sub.2 XVI 
wherein 
R.sub.1 is H or an acyl group of 1 to 7 C-atoms, 
A.sub.1 is D-Phe, which is optionally substituted in the phenyl ring by F, 
Cl, Br, NH.sub.2, CH.sub.3, or OCH.sub.3, especially in the 4 position, 
.beta.-D-naphthylalanine, D-Trp which optionally is substituted in the 5 
or 6 position by F, Cl, Br, NH.sub.2, or OCH.sub.3 and/or is substituted 
in position 1 by formyl or acetyl-, proline, 3,4-dehydroproline or 
D-pyroglutamine, 
B.sub.1 is D-Phe optionally substituted by F, Cl, Br, NH.sub.2, CH.sub.3 or 
CH.sub.3 O in the phenyl ring, D-.alpha.-methylphenylalanine, which is 
optionally substituted in the 4 position, by Cl, or 
.beta.-D-Naphthylalanine. 
C.sub.1 is D-Trp optionally substituted in position 5 or 6 by F, Cl, Br, 
NH.sub.2 and/or OCH.sub.3 and/or in position 1 by HCO or CH.sub.3 CO, 
.beta.-D-naphthylalanine, 3-D-pyridylalanine, D-Tyr, or Phe optionally 
substituted by F, Cl, Br, NH.sub.2, CH.sub.3, or CH.sub.3 O, 
D.sub.1 is Ser, 
E.sub.1 is Tyr, or phenylalanine optionally substituted by Cl, Br, 
NH.sub.2, CH.sub.3 or CH.sub.3 O, in the phenyl ring, 
F.sub.1 is D-Phe, optionally substituted in the phenyl ring by F, Cl, Br, 
NH.sub.2, CH.sub.3 or CH.sub.3 O, D-Trp optionally substituted in position 
5 or 6 by F, Cl, Br, NH.sub.2 or CH.sub.3 O and/or in position 1 by formyl 
or acetyl, D Tyr, .beta.-D-naphthylalanine, D-Leu, D-Ile, D-Nle, DVal, 
D-Ser (OtBu), D-Arg, optionally dialkylated with (C.sub.1-6)alkyl or 
(C.sub.5-6)cycloalkyl, D-homoarginine, optionally dialkylated with 
(C.sub.1-6)alkyl or (C.sub.5-6)cycloalkyl, D-His, D-His(Bzl), D-Lys, or 
D-Orn, both optionally dialkylated with (C.sub.1-6)alkyl or 
(C.sub.5-6)cycloalkyl, D-Phe (p---NH.sub.2) or 
.alpha.-p-aminocyclohexylalanine, 
G.sub.1 is Leu, Nle, Nva, N-.alpha.-methylleucine, Trp, Phe, Met or Tyr, 
H.sub.1 is Arg, Lys or Orn which optionally is substituted by 
(C.sub.1-6)alkyl or (C.sub.5-6)cycloalkyl, 
I.sub.1 is Pro, hydroxyproline, or 3,4-dehydroproline, and 
K.sub.1 is D-Ala. 
If desired E.sub.1 and F.sub.1 may be replaced by the other D.sub.1 and 
K.sub.1, if desired may be Cys which are linked by a S--S-- bridge. 
If desired one of D.sub.1 and K.sub.1 is Asp or Glu and the other is Orn, 
diaminoproponic acid or diaminobutyric acid and wherein the residue 
D.sub.1 and K.sub.1 are linked by an amide bridge. 
Preferred significances are: 
R.sub.1 =acetyl or formyl 
A.sub.1 =D-Phe, D-Phe (p-Cl), .beta.-D-naphthylalanine, 3,4-dehydroproline, 
B.sub.1 =D-Phe optionally substituted as indicated above 
C.sub.1 =D-Trp optionally substituted as indicated above 
D.sub.1 =Ser 
Optional substitution is preferably mono substitution. 
(i) E.sub.1 =Tyr or Phe optionally substituted as indicated above, when 
F.sub.1 =D-Phe or Lys or 
(ii) E.sub.1 =D-Phe or Lys when F.sub.1 =Tyr or Phe optionally substituted 
as defined above, 
G.sub.1 =Leu 
H.sub.1 =Arg 
I.sub.1 =Pro 
K.sub.1 =D-Ala 
In the above mentioned LHRH antagonists the sugar residue is preferably 
attached to the N-terminal amino group or to a free amino group in a side 
chain. 
The sugar derivatives preferably have the following structures wherein 
H.sub.2 N-LHRH antagonist denotes a LHRH antagonist of formula XVI: 
##STR44## 
In the above defined formulae XVIa to XVIf for simplicity only one sugar 
moiety is shown bound to a amino group. However, if desired more than one 
sugar moiety may be present. Preferably two such sugar moieties are 
present. 
The starting materials and the synthesis for the non-modified LHRH 
antagonists are for example described in EPA 81887 and 201260 A. 
Further preferred polypeptides are: 
a) oxytocin and vasopressin, as well as their derivatives, e.g. Lys.sup.8 
-vasopressin and Orn.sup.8 -vasopressin, 
b) Insulin 
c) Growth hormone releasing factor 
The starting materials and compounds of the invention may be produced by 
liquid phase or the solid phase synthesis. 
The compounds of the invention may be conveniently prepared by solid phase 
synthesis. 
We have found an especially convenient process for the production of 
peptide alcohols, which at the C-terminal end of the peptide chain bear 
two alcohol groups or one alcohol group and one thiol group. The process 
is especially suitable for the production of peptide alcohols which 
contain a C-terminal threoninol, serinol or cysteinol radical. 
Examples of suitable compounds include some of the somatostatin compounds 
described herein. 
Solid-phase peptide synthesis has proved to be an especially rapid and 
favourable process for the production of peptides, and has therefore 
become a generally conventional method. 
As is known, first of all an amino acid is bonded by its carboxyl group, 
forming an ester or amide group, to a hydroxyl group or amino group of an 
insoluble synthetic resin; then, the further amino acids are added onto 
this in the desired sequence, and finally the complete polypeptide is 
cleaved from the carrier resin. 
This synthesis operates without problems for normal polypeptides having 
C-terminal amino acids. However, polypeptide alcohols, which at the C-end 
bear an amino-alcohol instead of an amino acid, do not easily form a bond 
with carrier resins bearing OH-- or NH.sub.2 -- groups and/or are not so 
easily cleaved again when synthesis bas ended. 
The following have previously been proposed as possible solid-phase 
processes for the production of peptide alcohols: 
a) conventional preparation of the corresponding polypeptide containing at 
the C-end an amino acid (as the ester of a resin bearing OH groups) and 
subsequent cleavage by reduction, using boron hydrides, the carboxyl group 
being simultaneously converted into an alcohol function. (U.S. Pat. Nos. 
4,254,023/4). 
b) Addition of the terminal amino alcohol as ether to a hydroxymethyl 
resin, using carbonyl diimidazole, and finally, after synthesis of the 
peptide, cleavage using HCl/TFA or HBr/TFA (Kun-hwa Hsieh and G. R. 
Marshall, ACS National Meeting, New Orleans, 21-25. 3. 1977). 
However, these methods both require drastic cleavage conditions. 
We have found that the cleavage of the peptide from the resin, whilst 
simultaneously forming the C-terminal peptide alcohol, is carried out 
under mild conditions if the C-terminal amino-alcohol is joined to the 
resin by an acetal bond. 
In accordance with the invention, the peptide alcohol which at the 
C-terminal end of the peptide chain bears 2 alcohol groups or one alcohol 
group and one thiol group is produced by acid hydrolysis of an acetal of 
the peptide alcohol and a polymer resin bearing formylphenyl groups. This 
is referred to as the synthesis of the invention. 
The reaction may be illustrated schematically as follows: 
##STR45## 
wherein P is the residue of an insoluble synthetic resin 
Z is a direct bond or a residue which joins the resin with the (acetalised) 
formylphenyl group 
X is O or S 
R.sub.1 is hydrogen or methyl, and 
Y is the residue of a peptide alcohol which e.g. may bear protecting 
groups, 
wherein the optionally acetalised CHO-- group is located in the m- or 
p-position to the radical Z. For simplicity, in formulae I and II of the 
above scheme, only one substitution group was indicated on the resin; it 
should however be clear that a number of such groups are bonded to a 
molecule of the resin polymer. Cleavage of the peptide alcohol from the 
resin by hydrolysis of the acetal group takes place as mentioned above 
under acidic conditions, e.g. with diluted trifluoroacetic acid. 
Hydrolysis can be effected at room temperature. 
If Z in formula I.sub.r is a direct bond, the phenyl radicals bearing 
acetal groups are directly bonded to the polymer residue and belong to the 
polymer. Examples of such compounds of formula I.sub.r are the acetals of 
a formylated polystyrene resin (in formula I.sub.r, P is then a 
polyethylene chain). 
If Z is a residue, then this residue contains a group which is the result 
of a reaction of a reactive group, that is directly or indirectly bonded 
to the polymer, with another reactive group, that is directly or 
indirectly bonded to the (acetalised) formylphenyl group. The radical Z 
may be represented for example by the following formula IIIr: 
EQU -(D).sub.p -Q.sub.1 -Q.sub.2 -(E).sub.q - IIIr 
wherein 
Q.sub.1 =the residue of a reactive group which is bonded to the polymer 
Q.sub.2 =the residue of a reactive group which is bonded to the 
(acetalised) formylphenyl group 
D=a residue which joins the group Q.sub.1 with the polymer 
E=a residue which joins the group Q.sub.2 with the (acetalised) 
formylphenyl group 
p and q, independently of one another, are 0 or 1. 
The Q.sub.1 -Q.sub.2 group is preferably an ester or amide group, 
especially a carbonamide group. Q.sub.1 is preferably NH and Q.sub.2 is 
preferably CO. 
D and E, independently of one another, are for example alkylene or 
alkyleneoxy radicals having 1 to 5 C-atoms. Examples of such compounds of 
formula Ir, wherein Z is a residue of formula IIIr, are compounds wherein 
P-D-Q.sub.1 is the residue, of an aminomethylated polystyrene resin and 
the residue 
##STR46## 
is a residue of formula IVr 
##STR47## 
wherein R=hydrogen or methyl and 
m=0 or 1, 
whereby the acetal group is again located in m- or p-position. 
In this case, Z is thus 
##STR48## 
P is polystyrene. 
Radical IVr is preferably 
##STR49## 
Instead of the aminomethylated polystyrene, other polymers can also be 
used, especially those having free NH.sub.2 groups, e.g. polyacrylamides 
bearing aminoethyl groups. 
As mentioned above, the acetalised formylphenyl group is preferably bonded 
to the polymer by an amide bond. This ensures that the bonding of the 
acetalised formylphenyl radical to the resin during synthesis of the 
polypeptide and during cleavage is stable, and that cleavage occurs on the 
acetal bond as desired, so that on the one hand the peptide alcohol is 
generated and on the other hand the formylphenyl radical remains on the 
resin. 
If desired, the peptide alcohol can be attached further away from the resin 
by incorporating so-called spacers between the reactive groups of the 
polymer (especially amino groups) and the reactive groups of the 
acetalised formylphenyl derivative (especially carboxyl groups). For 
certain reactions on the polypeptide alcohol, this may advantageously be 
before cleavage (e.g. oxidation of cystein radicals). In this case, the 
radical D or E in formula IIr additionally contains the spacer and Q.sub.1 
or Q.sub.2 is the reactive radical of the spacer. 
The spacer used can be for example a .omega.-aminocarboxylic acid, such as 
.epsilon.-aminocaproic acid. 
In a specific case, when using aminomethylated polystyrene, a radical of 
formula IVr and .epsilon.-aminocaproic acid as the spacer, Z is 
##STR50## 
The compounds of formula I.sub.r can be produced by methods which are usual 
in solid-phase technology, starting with a compound of formula Vr 
##STR51## 
wherein A is a-protecting group of the amino function and the acetal group 
is in m- or p-position to the radical Z. For this purpose, first of all 
the protecting group A is cleaved and then the free amino group is reacted 
with the next N-protected amino acid etc., until all the amino acids have 
been added onto the resin in the sequence corresponding to the desired 
peptide alcohol. 
The amino protecting groups to be chosen for the amino acids used or for 
the amino alcohol must be those which are cleaved under non-acidic 
conditions, since under acidic conditions hydrolysis of the acetal group 
takes place. The CF.sub.3 CO-- or the FMOC-- group 
(9-fluorenylmethyloxycarbonyl) can be used e.g. as such amino protecting 
groups. These protecting groups are cleaved in a basic medium in a manner 
which is usual for peptide chemistry. 
Only protecting groups in the side chains and the amino protecting group of 
the last administered amino acid may be acid labile and then are 
simultaneously split off from the resin with the regeneration of the 
peptide alcohol. 
Preferably the Boc groups present as a protecting group. 
As bases are preferably used KOH or piperidine or NaBH.sub.4. 
The building up of the peptide chain may be effected in conventional manner 
from a peptide moiety having free amino groups and an amino acid with free 
or activated carboxyl groups. 
The reaction may be effected with the addition of e.g. hydroxybenzotriazol 
and dicyclohexylcarbodiimide. 
Compounds of formula Vr may be produced for example by 
a) reacting a resin carrying an aldehyde group of formula II.sub.r 
##STR52## 
wherein the CHO group is in the m or p position to the Z substituent, with 
an N-protected amino alcohol of formula 
EQU HX--CHR.sub.1 --CH(NHA)--CH.sub.2 OH 
optionally in activated form, or 
b) reacting a resin having the formula 
##STR53## 
with a compound of formula VIr 
##STR54## 
wherein the acetal group is in the m or p position to the Q.sub.1 
'--(E).sub.q -- group and 
Q.sub.1 ' and Q.sub.2 ' are two reactive groups which react together to 
form a Q.sub.1 -Q.sub.2 bridge. 
The acetalization of process a) may be effected in the presence of an acid 
as catalyst. Suitable acids include p-toluene sulphonic acid and 
p-trifluoromethylsulphonic acid. 
If desired a trimethylsilyl group may be used as a protecting group for a 
free alcohol. 
The esterification process b) may be effected under very mild conditions, 
e.g. by reaction of a carboxylic acid derivative with an OH or NH.sub.2 
group carrying polymer. 
The compounds of formula VIr may be produced by acetylation of a compound 
of formula 
##STR55## 
with a compound of formula 
EQU HX--CHR.sub.1 --CH(NHA)--CH.sub.2 OH 
The acetylisation may be effected as for process a). 
During the building up and the splitting off of the peptide alcohol from 
the resin, further reactions may be effected, e.g. removal of protecting 
groups, e.g. S-protecting groups, or oxidation of cystein radicals. 
Such reactions may be effected after splitting off of the peptide alcohol 
in the liquid phase. 
According to the synthesis of the invention pharmacologically active and 
other peptides which on the C-end contain 2 alcohol groups or an alcohol 
and a thiol group may be simply produced. 
In the following examples, all temperatures are given in degrees celsius 
and the [.alpha.].sub.D.sup.20 values are uncorrected. The following 
abbreviations are used: 
AcOH=acetic acid 
Boc=tert. butyloxycarbonyl 
Bu.sup.t =tert. butyl 
DCCI=dicyclohexylcarbodiimide 
DMF=dimethyl formamide 
Fmoc=9-fluorenylmethoxycarbonyl 
MeOH=methanol 
NEt.sub.3 =triethylamine 
Thr-ol=threoninol radical=CH.sub.3 --CHOH--CH(CH.sub.2 OH)--NH-- 
TFA=trifluoroacetic acid 
HOBT=N-hydroxybenzotriazole 
hpGRF=human pancreatic growth hormone releasing factor 
HOSu=N-hydroxy-succinimide 
All peptides are obtained as polyacetates-polyhydrates with a peptide 
content of 70 to 90%, 
HPLC analysis shows that the peptides contain less than 5% of other 
peptides. 
The factor "F" mentioned in the following examples shows the peptide 
content in the products obtained (F=1 conforms with 100% peptide content). 
The difference up to 100% [(1-F).times.100] consists of acetic acid and 
water. 
All sugars have the .alpha.-configuration unless otherwise stated. 
Deoxy=Desoxy.

EXAMPLE 1 
##STR56## 
3 ml of trifluoroacetic acid (100%) are added to 400 mg of 
##STR57## 
and kept at room temperature until all the starting material has dissolved 
(5 minutes). After adding 20 ml of diisopropylether, the title compound is 
precipitated and subsequently filtered off and washed with 
diisopropylether. The title compound is purified by chromatography on 
silica gel (eluant: CHCl.sub.3 /MeOH/HOAc/H.sub.2 O 7/3/0.5/0.5) and is 
isolated as a white lyophilisate. [.alpha.].sub.D.sup.20 : -31.3.degree. 
(c=0.52 in HOAc 95%). F: 0.88 
The starting product may be produced as follows: 
a) 
##STR58## 
2.25 g of di-tert.butyl-percarbonate, dissolved in 30 ml of DMF, are slowly 
added in drops at room temperature to a solution of 10 g of 
##STR59## 
in 100 ml of DMF. After two hours at room temperature, the solvent is 
drawn off-.under-vacuum, and 200 ml of diisopropylether are added to the 
residue. The deposit which is being formed is filtered off, washed with 
diisopropylether and dried. The crude product is purified by 
chromatography over silica gel (eluant: CH.sub.2 Cl.sub.2 /MeOH 9/1) and 
is then isolated as a white amorphous powder. [.alpha.].sub.D.sup.20 : 
29.8.degree. (C=1.28 in DMF) 
b) 
##STR60## 
2 g of D-(+)-glucose and 0.5 g of the end product of stage a) are 
dissolved in 20 ml of MeOH/HOAc 9/1 (v/v), and kept at 
60.degree.-70.degree. C. for three hours. After concentration by 
evaporation, the product is taken up in a little methanol, and the title 
compound is precipitated with diisopropylether. It is purified by 
chromatography over silica gel (eluant: CH.sub.2 Cl.sub.2 /MeOH 9/1). An 
amorphous powder is obtained. [.alpha.].sub.D.sup.20 =12.0.degree. (c=1.04 
in DMF) 
The following compounds (all as acetates) were produced analogously to 
example 1 (in these compounds, SMS denotes the polypeptide radical 
##STR61## 
EXAMPLE 2 
N.sup..alpha. -[.alpha.-glucosyl(1-4)-dioxyfructosyl)-SMS 
##STR62## 
starting with D(+)-maltose instead of D-glucose [.alpha.].sub.D.sup.20 
=-7.9.degree. (c=0.71 in AcOH 95%) F: 0.91 
EXAMPLE 3 
N.sup..alpha. 
-[.alpha.-glucosyl(1-4)-.alpha.-glucosyl(1-4)-.beta.-deoxyfructosyl-SMS 
##STR63## 
starting with maltotriose instead of D-glucose [.alpha.].sub.D.sup.20 
=+11.3.degree. (c=0.71 in 95% AcOH) 
EXAMPLE 4 
N.sup..alpha. -fructofuranuronic Acid-SMS 
##STR64## 
starting with D-glucuronic acid instead of D-glucose 
[.alpha.].sub.D.sup.20 =-29.4.degree. (c=0.34 in 95% AcOH) 
EXAMPLE 5 
N.sup..alpha. -deoxysorbosyl-SMS 
##STR65## 
starting with D (+)-galactose instead of D-glucose [.alpha.].sub.D.sup.20 
=-30.4.degree. (c=0.50 in 95% AcOH) 
EXAMPLE 6 
N.sup..alpha. -[O-.beta.-D-glucosyl-(1-4)-deoxyfructosyl]-SMS 
##STR66## 
starting with D(+)-cellobiose [.alpha.].sub.D.sup.20 =-28.1.degree. 
(c=0.47 in 95% AcOH) 
EXAMPLE 7 
N.sup..alpha. -L(-)-deoxyfructosyl-SMS 
##STR67## 
starting with L(-)-glucose instead of D(+)-glucose [.alpha.].sub.D.sup.20 
=-20.degree. (c=0.46 in 95% AcOH) 
EXAMPLE 8 
N.sup..alpha. -[O-.beta.-D-glucosyl-(1-6)-deoxyfructosyl]-SMS 
##STR68## 
starting with gentiobiose instead of D-glucose [.alpha.].sub.D.sup.20 
=23.5.degree. (c=0.46 in 95% AcOH) F: 0.76 
EXAMPLE 9 
N.sup..alpha. -[O-.beta.-D-galactosyl-(1-4)-deoxyfructosyl]-SMS 
##STR69## 
starting with D(+)-lactose instead of D-glucose [.alpha.].sub.D.sup.20 
=-29.3.degree. (c=0.55 in 95% AcOH) 
EXAMPLE 10 
N.sup..alpha. -(O-.alpha.-galactosyl-(1-6)-deoxyfructosyl)-SMS 
##STR70## 
starting with melibiose instead of D-glucose [.alpha.].sub.D.sup.20 
=+8.4.degree. (c=0.5 in 95% AcOH) F: 0.76 
EXAMPLE 11 
[N-(1-deoxy-D-fructosyl)-Tyr.sup.3 ]-SMS 
##STR71## 
starting with Tyr.sup.3 -SMS instead of SMS [.alpha.].sub.D.sup.20 
=-32.2.degree. (c=0.9 in 95% AcOH) F: 0.87 
EXAMPLE 12 
[N-(.alpha.-D-Glucopyranosyl-(1-4)-1-deoxyfructosyl), Tyr.sup.3 ]-SMS 
##STR72## 
starting with D(+)-maltose instead of D-glucose and Tyr.sup.3 -SMS instead 
of SMS [.alpha.].sub.D.sup.20 =-4.7.degree. (c=1.0 in 95% AcOH) F=0.81 
EXAMPLE 13 
##STR73## 
starting with D-glucoheptose instead of D-glucose [.alpha.].sub.D.sup.20 
=-12.9.degree. (c=1.0 in 95% AcOH) 
EXAMPLE 14 
##STR74## 
starting with D(+)-glucose and SMS, which does not have a protecting group 
on the .epsilon.-NH.sub.2 group of lysine [.alpha.].sub.D.sup.20 
=-42.4.degree. (c=0.37 in 95% AcOH) F=0.83 
EXAMPLE 15 
##STR75## 
starting with glucoheptose and SMS which does not have a protecting group 
on the .epsilon.-NH.sub.2 -- group of the lysine [.alpha.].sub.D.sup.20 
=-9.3+ (c=0.41 in 95% AcOH) F=0.84 
EXAMPLE 16 
Fructosyl-6-phosphat-SMS 
##STR76## 
starting from D-glucose-6-phosphate instead of D-glucose 
[.alpha.].sub.D.sup.20 =-19.5.degree. (c=1.0 in 95% AcOH) F=0.89 
EXAMPLE 17 
##STR77## 
starting from D-ribose instead of D-glucose [.alpha.].sub.D.sup.20 
=-31.8.degree. (c=1.0 in 95% AcOH) 
EXAMPLE 18 
N.sup..alpha. -deoxyfructosyl-(D)Phe-Cys[COC(CH.sub.3).sub.3 
]-Phe-(D)-Trp-Lys-Thr-Cys [COC(CH.sub.3).sub.3 ]-Thr-ol 
a) 
##STR78## 
0.58 g of the compound of example 1 in 10 ml of DMF are mixed with 0.08 mi 
of NEt.sub.3, then with 0.12 ml of (BOC).sub.2 O. The mixture is stirred 
for ca. 15 hours at room temperature, concentrated under vacuum and 
agitated with ether. The precipitated product is filtered off. The residue 
is dissolved with a little MeOH, then the product is precipitated by 
adding H.sub.2 O. The product is filtered, washed with a little H.sub.2 O, 
dried and the title compound is obtained. [.alpha.].sub.D.sup.20 
=+14.5.degree. (c=0.7 in DMF) 
b) N.sup..alpha. 
-deoxyfructosyl-(D)-Phe'-Cys-Phe-(D)Trp-Lys(BOC)-Thr-Cys-Thr-ol 
0.51 g of the end compound of stage a) in a mixture of 10 ml of dioxane and 
2 ml of NEt.sub.3 /AcOH buffer pH 8.6 under argon is mixed with a total of 
0.4 g of dithioerythritol. The mixture is stirred for ca. 15 hours at room 
temperature and concentrated under vacuum. The precipitated product is 
centrifuged off. The residue is washed with a little H.sub.2 O, then 
vacuum-dried. The title compound is obtained. [.alpha.].sub.D.sup.20 
=+3.8.degree. (c=0.8 in DMF) 
c) N.sup..alpha. -deoxyfructosyl-(D)Phe'-Cys[COC(CH.sub.3).sub.3 
]-Phe-(D)Trp-Lys-(B)C)-Thr-Cys(COC(CH.sub.3).sub.3 ]-Thr-ol 
0.38 g of the end compound of stage b) are dissolved under argon in25 mi of 
N-methylpyrrolidone, then mixed at 0.degree. with 0.3 ml of 
N-methylmorpholine and 0.31 ml of pivaloylchloride, and stirred for ca. 16 
hours at 0.degree.. The product is agitated with ether/diisopropylether. 
The precipitated product is centrifuged off. The residue is dissolved with 
a little DMF and the product precipitated by adding MeOH and H.sub.2 O. 
The product is centrifuged. The residue is vacuum-dried and used further 
without further purification. 
d) N.sup..alpha. -deoxyfructosyl-(D)Phe-Cys[COC(CH.sub.3).sub.3 
]-Phe-(D)Trp-Lys-Thr-Cys[COC(CH.sub.3).sub.3 ]-Thr-ol 
The residue of stage c) is dissolved at 0.degree. in 5 ml of TFA/H.sub.2 O 
(9:1) and stirred for 15 minutes. The product is precipitated by adding a 
mixture of ether/10% 5n HCl/ether. The product is filtered, washed with 
ether and dried. The residue is purified by chromatography on silica gel 
in a mixture of CHCl.sub.3 /MeOH/AcOH/H.sub.2 O. Fractions which contain 
the desired product are combined, concentrated under vacuum whilst adding 
H.sub.2 O, then lyophilised. The title compound is obtained. 
[.alpha.].sub.D.sup.20 =-15.3.degree. (c=1.0 in 95% AcOH) F: 0.88 
EXAMPLE 19 
##STR79## 
2 g of D(-)-fructose and 1 g of 
##STR80## 
(produced as described in example 1a) are dissolved in 100 ml of MeOH/HOAc 
9/1 and kept at 65.degree. C. for 16 hours. After concentration by 
evaporation, the product is dissolved in a little methanol, and the crude 
product is precipitated with diisopropylether. The crude product thus 
obtained is used in the protecting group cleavage (BOC cleavage) without 
being purified. 
1 g of the crude product obtained is mixed with 20 ml of tri-fluoroacetic 
acid (100%) and kept at room temperature until the entire starting 
material has dissolved (5 minutes). By adding 200 ml of diisopropylether, 
the title compound is precipitated and subsequently filtered off and 
washed with diisopropylether. The title compound is purified by 
chromatography on silica gel (eluant: CHCl.sub.3 /MeOH/HOAc/H.sub.2 O 
7/3/0.5/0.5) and is isolated as a white lyophilisate. 
[.alpha.].sub.D.sup.20 =-6.7.degree. (c=0.3 in HOAc 95%) F: 0.73 
As second product the following 1:1 mixture of isomers having the inverse 
configuration at C.sub.2 of the carbohydrate moiety may be obtained: 
##STR81## 
EXAMPLE 20 
2-[Tyr.sup.3 -SMS]-2-dioxy-D-glucose 
##STR82## 
In analogy to example 19 starting from Tyr.sup.3 SMS instead of SMS the 
heading compound is produced. [.alpha.].sub.D.sup.20 =-2.9.degree. (c=1.0 
in 95% AcOH) F=0.95 
EXAMPLE 21 
Glucoronic Acid Amide of 
##STR83## 
170 mg of the glucuronic acid amide of 
##STR84## 
are treated with 3 ml of trifluoroacetic acid 100%) until a complete 
solution is obtained (5 minutes). The title compound is subsequently 
precipitated as the trifluoroacetate by adding 20 ml of diisopropylether, 
and after filtration, drying and subsequent chromatography on silica gel 
(eluant: CHCl.sub.3 /MeOH/HOAc/H.sub.2 O 7/3/0.5/0.5), the title compound 
is isolated in pure form as a white lyophilisate (acetate). 
[.alpha.].sub.D.sup.20 =-29.2.degree. (c=0.48 in HOAc 95%) 
The starting product may be produced as follows: 
A solution of 135 mg of DCCI in 2 ml of DMF is added to a solution, cooled 
to -30.degree. C., in DMF of 450 mg of 
##STR85## 
117 mg of glucuronic acid and 135 mg of HOBT. After 48 hours, with 
simultaneous thawing to room temperature, the resultant dicyclohexylurea 
is filtered off and the title compound is precipitated by adding 20 ml of 
diisopropylether. After filtration, drying and chromatography over silica 
gel (eluant: CH.sub.2 Cl.sub.2 /MeOH 9/1), the title compound is isolated 
in pure form [.alpha.].sub.D.sup.20 =+16.7.degree. (c=0.50 in DMF) 
EXAMPLE 22 
Quinic Acid Amide of 
##STR86## 
The title compound Was obtained analogously to example 21, starting with 
L(-)-quinic acid. [.alpha.].sub.D.sup.20 =-50.degree. (c=0.44 in 95% AcOH) 
F: 0.97 
EXAMPLE 23 
Sialic Acid Amide of 
##STR87## 
The title compound was obtained analogously to example 21, starting with 
sialic acid. [.alpha.].sub.D.sup.20 =-60.8.degree. (c=0.6 in 95% AcOH) F: 
0.95 
EXAMPLE 24 
##STR88## 
250 mg of 
##STR89## 
are dissolved in 10 ml of methanol and adjusted to a pH of 10 with a few 
drops of 1N NaOCH.sub.3 solution in methanol. After reacting for 15 
minutes, the solution is neutralised with an ion exchanger (e.g. 
AMBERLYST.RTM. 15, H.sup.+), and the ion exchanger is filtered off. The 
filtrate is concentrated and the residue treated for 5 minutes with 3 ml 
of trifluoroacetic acid. The title compound is precipitated as the 
trifluoroacetate by adding 20 ml of diisopropylether, and is isolated in 
pure form as a white lyophilisate after filtration, drying and 
chromatography on silica gel (eluant: CHCl.sub.3 /MeOH/HOAc/H.sub.2 O 
7/3/0.5/0.5). [.alpha.].sub.D.sup.20 =-39.2.degree. (c=0.60 in HOAc 95%) 
F: 0.91 
The starting product may be produced as follows: 
a) tetra-O-acetyl-O-.beta.-D-glucosyl-glycolic acid benzylester 
2.5 g of molecular sieve 4 .ANG., powder are added to a solution of 830 mg 
of glycolic acid benzylester in 50 ml of CH.sub.2 Cl.sub.2, and after 
adding 2.8 g of silver trifluoromethane sulphonate, a solution of 4.1 g of 
acetobromoglucose in 50 ml of CH.sub.2 Cl.sub.2 is added in drops. After 
15 minutes, the reaction is stopped with 4 ml of pyridine, the solid 
constituents are filtered off, and the filtrate is shaken out with 10% 
NaHSO.sub.4 solution. The title compound is isolated in pure form after 
chromatography over silica gel (eluant: CH.sub.2 Cl.sub.2 /MeOH 99/1). 
[.alpha.].sub.D.sup.20 =-22.4.degree. (c=1.7 in CHCl.sub.3) 
b) tetra-O-acetyl-O-.beta.-D-glucosyl-glycolic acid 
800 mg of tetra-O-acetyl-O-.beta.-D-glucosyl-glycolic acid benzylester are 
dissolved in 40 ml of ethanol/water 1/1 (v/v), and mixed with 400 g of 
palladium/active charcoal 10%. Hydrogenation on "R-APATUS" at 50 PSI 
produces the title compound, which is isolated in crystalline form after 
filtration and concentration under vacuum. [.alpha.].sub.D.sup.20 
=-35.5.degree. (c=1.03 in MeOH) 
c) 
##STR90## 
To a solution of 81 mg of tetra-O-acetyl-O-.beta.-D-glucosyl-glycolic acid, 
225 mg of 
##STR91## 
and 45 mg of HOBT in 2 ml of DMF, cooled to -30.degree. C., are added 45 
mg of DCCI, dissolved in 1 ml of DMF. After 48 hours and after thawing to 
room temperature, the resultant dicyclohexylurea is filtered off, and the 
title compound is precipitated from the filtrate by adding 20 ml of 
diisopropylether. 
The following compounds were also produced analogously to example 24 (in 
these, SMS denotes the radical 
##STR92## 
EXAMPLE 25 
N.sup..alpha. -(O-.beta.-D-galactosyl-oxyacetyl)-SMS 
##STR93## 
[.alpha.].sub.D.sup.20 =-37.5.degree. (c=1 in 95% AcOH) F: 0.95 
EXAMPLE 26 
N.sup..alpha. -(O-.beta.-cellobiosyl-oxyacetyl)-SMS 
##STR94## 
[.alpha.].sub.D.sup.20 =-32.5.degree. (c=1 in 95% AcOH) F: 0.91 
EXAMPLE 27 
N.sup..alpha. -(O-.beta.-(D)-glucosyl-oxyisobutyryl)-SMS 
##STR95## 
[.alpha.].sub.D.sup.20 =-32.9.degree. (c=1 in 95% AcOH) F: 0.93 
EXAMPLE 28 
N.sup..alpha. -(O-.alpha.-(D) -glucosyl-s-(L)-oxyisovaleryl)-SMS 
##STR96## 
[.alpha.].sub.D.sup.20 =-44.3.degree. (c=1 in 95% AcOH) F: 1.00 
EXAMPLE 29 
[N-acetylmuramyl-(D)Phe.sup.1 ] -SMS 
##STR97## 
[.alpha.].sub.D.sup.20 =-15.4.degree. (c=0.13 in 95% AcOH) F: 0.9 
EXAMPLE 30 
.beta.-D-Glucosyl-thiocarbamyl-SMS 
##STR98## 
620 mg E-Fmoc-SMS in 50 ml CH.sub.3 CN/H.sub.2 O 3:] are treated with 0.45 
ml triethylamine. 272 mg 
2,3,4,6-tetra-O-acetyl-.beta.-D-glucosyl-isothiocyanat are added and the 
mixture maintained at room temperature for 1 hour. The mixture is 
evaporated in a vacuum and the residue is taken up in a little methanol 
and treated with diisopropylether where: upon the product precipitate in 
practically pure form. 
To split off the Fmoc group and the acyl group, the product in 50 ml 
absolute methanol is treated with a catalytic quality of IN NaOCH.sub.3 in 
methanol. After 30 minutes time the reaction had been completed (by tlc), 
the mixture is neutralized with 1% acetic acid and evaporated in a vacuum. 
The residue is taken up in water and extracted with ethyl acetate. The 
aqueous phase is lyophilized. The residue is purified over silica gel and 
desmineralized over e.g. Duolite. The title compound is obtained as a 
lyophilisate. [.alpha.].sub.D.sup.20 =-48.5.degree. (c=1, 95% AcOH) F=1 
The starting material E-Fmoc-SMS may be produced as follows: 
5 g SMS acetate and 5 g NaHCO.sub.3 in 100 ml DMF/H.sub.2 O 3:1 are treated 
with 1.6 g Fmoc-HOSu. After an hour at room temperature, the mixture is 
diluted with 400 ml H.sub.2 O and extracted with 250 ml ethyl 
acetate/methanol 95:5. The organic phase is dried with Na.sub.2 SO.sub.4 
and concentrated. After column chromatography over silica gel the starting 
material is obtained as an amorphous substance. [.alpha.].sub.D.sup.20 
=24.3.degree. (c=1.13 DMF) 
The following products may be obtained in analogous manner to that 
described in Example 30. 
EXAMPLE 31 
Cellobiosylthiocarbamyl-SMS 
Starting from octa-acetyl-cellobiosyl-isothiocyanate 
##STR99## 
[.alpha.].sub.D.sup.20 : -4.3.degree. (c=1 in AcOH) F=0.87 
EXAMPLE 32 
.beta.-D-Glucosylcarbamoyl-SMS 
Starting from 2,3,4,6-tetra-O-acetyl-.beta.-D-glucosyl-isocyanate 
##STR100## 
[.alpha.].sub.D.sup.20 : -39.9 (c=1 in 95% AcOH) F=0.81 
EXAMPLE 33 
Cellobiosylcarbamoyl-SMS 
Starting from octa-acetyl-cellobiosyl-isocyanate 
##STR101## 
[.alpha.].sub.D.sup.20 =-37.9.degree. (c=1 in 95% AcOH) F=0.85 
EXAMPLE 34 
1-Deoxy-D-sorbityl-SMS 
##STR102## 
0.5 mg of the title compound of example 1 in 50 ml methanol is treated 
first with NaBH.sub.4, then 5% acetic acid under conditions such that the 
pH does not increase beyond 7. Total use of NaBH.sub.4 is about 10 
equivalents. 
After the complete reaction has occurred (4-5 hours) the mixture is treated 
with acetic acid to destroy excess NaBH.sub.4. The mixture is concentrated 
under a vacuum. The residue is desmineralized with e.g. Duolite and 
purified over silica gel. The main compound besides 
1-desoxy-D-mannityl-SMS is the title compound which is produced as a 
lyophilisate. [.alpha.].sub.D.sup.20 =-17.6.degree. (c=1 in 95% AcOH) 
F=0.82 
EXAMPLE 35 
.alpha.-D-glucosyl(1-4)deoxysorbityl-SMS 
In analogous manner to that described in Example 34 starting from the title 
compound of Example 2 the following compound is produced 
##STR103## 
[.alpha.].sub.D.sup.20 =+1.6.degree. (c=1 in AcOH) F=0.9 
EXAMPLE 36 
1,2-dideoxy-sorbityl-SMS 
##STR104## 
0.55 g Boc-SMS in 30 ml dioxane /H.sub.2 O 3:7 are treated with 50 mg 
NaBH.sub.3 CN. 250 mg 2-deoxy-D-glucose are added. The pH of the mixture 
is adjusted to 7 with: 0.1 ml HCl and heated to 100.degree. C. for 6 
hours, The mixture is cooled, freezed and lyophilized. The residue is 
taken up in ethyl acetate (50 ml) and shaken with water. The organic phase 
is dried and evaporated in a vacuum. The Boc group is split off in 
conventional manner with TFA. The product is purified over silica gel and 
desmineralized e.g. over Duolite to give the title compound. 
[.alpha.].sub.D.sup.20 =25.degree. (c=1.95% HOAc) F=0.83 
In analogous manner compounds of the foregoing examples 34 (starting from 
glucose) and 35 (starting from maltose) may be produced. 
EXAMPLE 37 
N.sup..alpha. 
-isocaproyl-des(1-4)-[Ala.sup.7,N.epsilon.-(1-deoxyfructosyl)-Lys.sup.11,1 
8 ]salmon Calcitonin 
##STR105## 
10.3 g of N.sup..alpha. -isocaproyl-des(1-4)-[Ala.sup.7 ]salmon calcitonin 
polyacetate and 1o8 g of D(+)-glucose are dissolved in a mixture of g4 ml 
of DMF and 6 ml of acetic acid. After 2 hours at 50.degree. C., the 
product is completely precipitated by adding ether, then filtered off by 
suction, washed with ether and vacuum-dried. Purification is effected by 
dissolving ca. 5-10 g of the product in water, adding the solution to a 
reversed-phase column 4.times.25 cm, C-18 on silica-gel and 
chromatographing with a gradient of water and 0-80% of a solvent mixture 
comprising 38 parts of water, 60 parts of acetonitrile and 2 parts of 85% 
phosphoric acid. The fractions which contain the pure product are 
combined, filtered over a column of ca. 100 ml of a slightly basic ion 
exchanger in acetate form and washed with water. The filtrate is 
lyophilised and the title compound is obtained as the polyacetate, 
polyhydrate. [.alpha.].sub.D.sup.20 =-34.8.degree. (c=0.73 in CH.sub.3 
COOH 95%) F: 0.93 FAB mass spectroscopy 3407 (MH.sup.+) 
The N.sup..alpha. 
-isocaproyl-Ser-Thr-Ala-Val-Leu-Gly-Lys-Leu-Ser-Gln-Glu-Leu-His-Lys-Leu-Gl 
n-Thr-Tyr-Pro-Arg-Thr-Asn-Thr-Gly-Ser-Gly-Thr-Pro-NH.sub.2 used as the 
starting product may be produced as follows: 
a) N.sup..alpha. 
-isocaproyl-Ser(Bu.sup.t)-Thr(Bu.sup.t)-Ala-Val-Leu-OCH.sub.2 
-phenyl-(p)OCH.sub.2 -co-(polystyrene-1%-divinylbenzene) 
1 g of p-hydroxymethyl-phenoxymethyl-co(polystyrene-1%-divinylbenzene) is 
left to swell in dimethylformamide/methylene chloride 1:4 (v/v), filtered 
off by suction and mixed with a solution of 0.74 g of Fmoc-leucine and 
0.19 g of i-hydroxybenzotriazole in 5 ml of the above-mentioned solvent 
mixture. 0.43 g of dlcyclohexylcarbodiimide and 85 mg of 
4-dimethylaminopyridine, each in 5 ml of the same solvent mixture, are 
added whilst stirring. The mixture is stirred for 16 hours at 20.degree., 
filtered off by suction and washed with the solvent mixture, then with 
dimethylformamide. Fmoc-Leu-OCH.sub.2 -phenyl-(p)-OCH.sub.2 
-co(polystyrene-1%-divinylbenzene) is obtained. 
The N.sup..alpha. -Fmoc group is split from the Fmoc-Leu-OCH.sub.2 
-phenyl-(p)OCH.sub.2 -co(polystyrene-1%-divinylbenzene) (1.56 g 
corresponding to 0.7 mMol) by treating with piperidine (20% v/v) in DMF 
for 10 minutes. This is washed well with DMF, and then 0.71 g of 
Fmoc-Val-OH, 0.28 g of 1-hydroxybenzotriazole and 0.32 ml of 
diisopropylcarbodiimide, each dissolved in 5 ml of DMF, are added. After 
45 minutes, the mixture is filtered by suction, and the peptide resin is 
washed well with DMF. The splitting of the N.sup..alpha. -Fmoc group is 
repeated, as well as the coupling with the amino acid following in 
sequence, in the order given: Fmoc-Ala-OH (0.65 g) Fmoc-Thr(Bu.sup.t)-OH 
(0.83 g). and Fmoc-Ser(Bu.sup.t)-OH (0.80 g). In the latter reaction cycle 
(splitting of the Fmoc protecting group, acylation with protected amino 
acid) the amino acid derivative is replaced by isocaproic acid (0.41 g), 
the quantity of 1-hydroxy-benzotriazole is increased to 0.53 g and that of 
diisopropylcarbodiimide to 0.54 g, and coupling is effected for 15 hours. 
The protected peptide resin is washed well with DMF and methylene 
chloride, vacuum-dried at 40.degree. C. for 15 hours, and the protected 
peptide resin is obtained as a colourless powder. 
b) N.sup..alpha. -isocaproyl-Ser-Thr-Ala-Val.sup.-Leu-OH 
N.sup..alpha. -isocaproyl-Ser(Bu.sup.t)-Thr(Bu.sup.t)-Ala-Val-Leu-OCH.sub.2 
-phenyl-(p)OCH.sub.2 -co(polystyrene-1%-divinylbenzene) (1.0 g) is stirred 
in a mixture of trifluoroacetic acid (5 ml) and methylene chloride (5 ml). 
The product is filtered, washed with the same mixture (5 ml), then with 
methylene chloride, greatly concentrated under vacuum, and totally 
precipitated by adding ether. The deposit is washed well with ether, dried 
under vacuum over solid potassium hydroxide, and the title compound is 
obtained as a colourless, amorphous powder. 
c) N.sup..alpha. 
-isocaproyl-Ser-Thr-Ala-Val-Leu-Gly-Lys(Boc)-Leu-Ser-Gln-Glu(OBu.sup.t)-Le 
u-His-Lys(Boc)-Leu-Gln-Thr-Tyr-Pro-Arg-Thr-Asn-Thr-Gly-Ser-Gly-Thr-Pro-NH.s 
ub.2 
To a solution of N.sup..alpha. -isocaproyl-Ser-Thr-Ala-Val-Leu-OH (0.165 g) 
in DMF (7 ml) are added 
H-Gly-Lys(Boc)-Leu-Ser-Gln-Glu(OBu.sup.t)-Leu-His-Lys(Boc)-Leu-Gln-Thr-Tyr 
-pro-Arg-Thr-Asn-Thr-Gly-Ser-Gly-Thr-Pro-NH.sub.2 hydrochloride (0.59 g), 
3,4-dihydro-3-hydroxy-4-oxo-1,2,3-benzotriazine (0.017 g), 
dicyclohexylcarbodiimide (0.065 g) and sufficient 
N-ethyl-N,N-diisopropylamine for a sample of the reaction mixture on 
moistened pH paper to indicate a reaction of ca. pH 6. After 16 hours, the 
mixture is precipitated by adding ether, dried, and the title compound is 
obtained. 
d) N.sup..alpha. 
-isocaproyl-Ser-Thr-Ala-Val-Leu-Gly-Lys-Leu-Ser-Gln-Glu-Leu-His-Lys-Leu-Gl 
n-Thr-Tyr-Pro-Arg-Thr-Asn-Thr-Gly-Ser-Gly-Thr-Pro-NH.sub.2 
0.50 g of the partly protected peptide of stage c) are dissolved in a 
mixture of trifluoroacetic acid (50% v/v) and methylene chloride. After 1 
hour, 50 ml of ether which contains 0.6 mMol of HCl is added. The mixture 
is filtered, washed with ether and vacuum-dried. The product is purified 
by "reversed-phase" chromatography in a gradient of acetonitrile in 
H.sub.3 PO.sub.4 (2%). The combined fractions containing the pure 
substance are filtered over a basic ion exchanger in acetate form. The 
filtrate is lyophilised and the title compound is obtained as the 
polyacetate, polyhydrate. [.alpha.].sub.D.sup.20 =-32.2.degree. (c=0.3 in 
AcOH 95%) F=0.87 
EXAMPLE 38 
N.sup..alpha. -isocaproyl-des-(1-4)-[Ala.sup.7, N.sup..kappa. 
-(.alpha.-D-glucosyl-(1-4)-deoxyfructosyl)-Lys.sup.11,18 ]salmon 
Calcitonin 
##STR106## 
The corresponding di-N.sup..epsilon. -maltulosyl derivative is produced 
analogously to example 37 using D(+)-maltose monohydrate instead of D 
(+)-glucose. The reaction time at 50.degree. C. is lengthened to 15 hours. 
Isolation and purification are identical and the title compound is 
obtained as the polyacetate, polyhydrate, FAB mass spectroscopy: 3730.9 
(MH.sup.+) [.alpha.].sub.D.sup.20 =-1.52.degree. (c=0.16 in 95% AcOH) 
F=0.97 Analogously to example 37 the following compounds Ire prepared: 
EXAMPLE 39 
N.sup..alpha. -isocaproyl-[N.sup..epsilon. -(1-deoxyfructosyl)-Lys.sup.7 
]-salmon Calcitonin [5-32)amide 
##STR107## 
[.alpha.].sub.D.sup.20 =-40.0.degree. (c=0.27 in AcOH 95%) F=0.84 
EXAMPLE 40 
N.sup..alpha.,Lys.sup.11 -N.sup..epsilon.,Lys.sup.18 N.sup..epsilon. 
-tris-(1-deoxyfructosyl)-salmon Calcitonin 
##STR108## 
[.alpha.].sub.D.sup.20 =-5.3.degree. (c=0.38 in AcOH 95%) F=0.75 
EXAMPLE 41 
N.sup..alpha. -isocaproyl-des(1-4)-[N.sup..epsilon. 
-(1-deoxyfructosyl)-Lys.sup.7,11,18 ]salmon Calcitonin-(5-32)amide 
##STR109## 
[.alpha.].sub.D.sup.20 =-37.4.degree. (c=0.155 in 95% AcOH) F=0.84 
EXAMPLE 42 
N.sup..alpha. -quinoyl-[Ala.sup.7 ]salmon Calcitonin-(5-32)-amide 
##STR110## 
The title compound was produced analogously to example 21. 
[.alpha.].sub.D.sup.20 =-35.7.degree. (c=0.37 in AcOH 95%) F: 0.88 
EXAMPLE 43 
N.sup..alpha. -quinoyl-[Ala.sup.7 ]salmon Calcitonin-(4-32)-amide 
##STR111## 
The title compound was produced analogously to example 21. 
[.alpha.].sub.20.sup.D =-39.3.degree. (c=0.29 in 95% AcOH) F: 0.89 
The starting peptides [Ala.sup.7 ]-salmon calcitonin-[5-32]-amide and 
[Ala.sup.7 ]-salmon calcitonin-(4-32)-amide required for examples 42 and 
43 may be produced analogously to the starting material of example 37. 
EXAMPLE 44 
[N-deoxy-fructosyl-Cys.sup.1 ]-oxytocin 
##STR112## 
0.5 g of D(+)-glucose and 0.5 g of oxytocin are dissolved in 50 ml of 
MeOH/HOAc 9/1 and kept at 65.degree. C. for 3 hours, The solution is then 
concentrated by evaporation, chromatographed over silica gel and freed 
from salt over Duolite (H.sub.2 O/ethanol/HOAc gradient). A white 
lyophilisate is obtained. [.alpha.].sub.D.sup.20 =-23.degree. (c=0.32 in 
95% HOAc) F: 0.95 
EXAMPLE 45 
Ac-(D)-Phe(4-Cl)-(D)Phe(4-Cl)-(D)Trp-Ser-Tyr- 
##STR113## 
60 mg of 
Ac-(D)Phe(4-Cl)-(D)Phe(4-Cl)-(D)Trp-Ser-Tyr-(D)-Lys-Leu-Arg-Pro-(D)Ala-NH. 
sub.2 and 72 mg of D(+)glucose are dissolved in a mixture of 10 ml of MeOH 
and 1 ml of AcOH, and stirred for ca. 20 hours at 60.degree. C. The 
product is precipitated with ether and centrifuged off. The residue is 
dissolved in ca. 100 ml of H.sub.2 O, and the pH is adjusted to 8 with 
dilute NaOH. The product is adsorbed on a column of Duolite ES 861 and 
eluted with a gradient of H.sub.2 O.fwdarw.dioxane-H.sub.2 O-AcOH 
(60:40:3). Fractions containing the desired product are concentrated under 
vacuum, then lyophilised. The title compound is obtained. 
[.alpha.].sub.D.sup.20 =-25.degree. (c=0.5 in 95% AcOH) F: 0.83 
EXAMPLE 46 
N.sup..alpha.A1,N.sup..alpha.B1,N.sup..epsilon.B29 
-tris(1-deoxyfructosyl)-porcine-Insulin 
A suspension of 1 g (0.17 mmol) and 0.47 g (2.6 mmol) glucose in 10 ml 
dimethylformamide/acetic acid 9:1 are stirred for 1 hour at 60.degree. C. 
The solvent is removed at 30.degree. C. in a high vacuum. The residue is 
dissolved in 300 ml H.sub.2 O, adjusted to pH 7 and the mixture passed 
through a small desmineralising column (Duolite ES 861 2.5.times.15 cm). 
The glucose is eluted with water and the peptide by isopropanol/water 
ethyl acetate 59:39:2. 
The solvent is removed and the mixture lyophilised. The residue is taken up 
in 300 ml water and purified through reversed phase chromatography. 
(2.times.25 cm column, RP 18, 10 nm, Buffer 57 mmol NaClO.sub.4, 20 mmol 
triethylamine, 8.4 mmol phosphoric acid, pH 3 with 4N NaOH, Gradient 0-65% 
A-B. 
##STR114## 
The fractions containing the heading compound are collected, combined, 
concentrated, diluted with 300 ml water and passed through a 
desmineralising column as described above. Salts are eluted with water. 
The peptide is eluted with isopropanol/water/ethyl acetate 59:39:2. The 
appropriate fractions are combined and concentrated to give the heading 
compound. [.alpha.].sub.D.sup.20 =56.3.degree. (c=0.5 AcOH) F=0.88. 
EXAMPLE 47 
N.sup..alpha.,Lys.sup.12 -N.sup..epsilon.,Lys.sup.21 -N.sup..epsilon. 
-tris-(1-deoxyfructosyl)-(D[Ala.sup.2 ]-hpGRF-(1-29)-NH.sub.2 
##STR115## 
In analogous manner to example 37 starting from (DAla.sup.2 ]hpGRF--the 
title compound is produced. [.alpha.].sub.D.sup.20 =-5.6.degree. (c=0.2 in 
95% AcOH) F=0.82 
EXAMPLE 48 
N.sup..alpha.,N.sup..epsilon. -bis(1-deoxyfructosyl)Lys-vasopressin 
##STR116## 
A suspension of 118 mg (0.1 mmol) Lys.sup.8 -vasopressin and 360 mg (2 
mmol) glucose in 5 ml methanol/ethyl acetate 9:1 are stirred at 65.degree. 
for 2 to 4 hours. The solvent is removed under a vacuum. The residue is 
taken up in 30 ml water and the solution lyophilised. To remove the excess 
glucose the peptide (solution in 40 ml water at pH 7.3) is adsorbed on a 
Duolite column (1.5.times.10 cm). The glucose is eluted with water and the 
peptide with a mixture of isopropyl/water/ethyl acetate 59:39:2. The 
mixture is purified on a silica gel column (eluant 
chloroform/methanol/ethyl acetate/water 7:4:1:1). 
The fractions containing the heading compound, are concentrated and 
lyophilised to give the title compound. [.alpha.].sub.D.sup.20 
=-52.degree. (c=0.5 in 95% HOAC) F=0.84. 
EXAMPLE 49 
##STR117## 
The title compound is produced in analogous manner to example 45 starting 
from 
Ac-(D)Phe(pCl)-(D)Phe(pCl)-(D)-Trp-Ser-Lys-(D)Phe-Leu-Arg-Pro(D)Ala-NH.sub 
.2, acetate and D(+)glucose. [.alpha.].sub.D.sup.20 =-36.degree. (c=0.5 in 
95% AcOH) F=0.86 
EXAMPLE 50 
##STR118## 
The title compound is produced in analogous manner to example 45 starting 
from 
H-(D)Phe(pCl)-(D)Phe(pCl)-(D)-Trp-Ser-Tyr-(D)Phe-Leu-Arg-Pro-(D)Ala-NH.sub 
.2, acetate and D(+)glucose. [.alpha.].sub.D.sup.20 =-32.degree. (c=0.5 in 
95% AcOH) F=0.94 
EXAMPLE 51 
##STR119## 
200 mg 
H-(D)Phe(pCl)-(D)Phe(pCl)-(D)Trp-Ser-Tyr-(D)Lys-Leu-Arg-Pro-(D)Ala-NH.sub. 
2 and 520 mg D(+)glucose in DMF/AcOH 15:1 are stirred at 60.degree. C. for 
3 hours. The mixture is concentrated in a vacuum, precipitated with ether, 
filtered and dried. 
The residue is purified as follows: 
1) Adsorption on Duolite ES 861 and elution with a mixture of 
dioxan-H.sub.2 O-AcOH. 
2) Column chromatography on silica gel using as eluant CHCl.sub.3 
/AcOH/H.sub.2 O. 
3) Preparative HPLC ("Reversed phase") chromatography on an 
octadecyl-silica gel column. Elution with an acetonitrile gradient in 2% 
H.sub.3 PO.sub.4. 
Fractions which contain the heading compound are combined, filtered through 
a column containing a weakly basic ion exchanger in acetate form, 
concentrated and lyophilised to give the title compound. 
[.alpha.].sub.D.sup.20 =-22.6.degree. (c=0.5 in 95% AcOH) F=0.63 
The synthesis of the invention may be effected as follows: 
EXAMPLE S1 
Production of Octreotide (=SMS) 
1) Production of acetal anchor (N-CF.sub.3 CO-Threoninol acetal of 
p-formylphenoxy-acetic acid). 
105 g (1.0 mmol) L-Threoninol is added to 200 ml methanol which is stirred 
by a stream of nitrogen. A clear solution results. A solution of 200 ml 
trifluoro acetic acid methyl ester in 250 ml methanol is added to the 
mixture at 0.degree.. The mixture is maintained at a temperature of about 
10.degree. C. by cooling with an ice bath. 
After 1.5 hours no more free Threoninol is detectable in the mixture. 
Concentration at 40.degree. C. gives a white crystalline residue. 
The residue is dissolved in 200 ml ethyl acetate at 70.degree. C. and 
precipitated by the addition of hexane. The mixture is cooled to 0.degree. 
C., washed with hexane and dried at room temperature. N-trifluoroacetyl 
threoninol results. 
50.3 g (0.25 mol) of the resultant product is dissolved in 1.25 liters 
tetrahydrofuran and 75 ml of tri methyl-chlorosilane is added dropwise. 
Immediately thereafter a mixture of 70 ml triethylamine and 250 ml 
tetrahydrofuran is added. A white suspension results which is stirred for 
4 hours. The mixture is filtered and the filtrate evaporated at 40.degree. 
C. to give an oil. 
The oil is dissolved in 1.5 liters of methylene chloride and treated with 
portions of 90.4 g p-formyl-phenoxyacetic acid at room temperature. 
Portions of 9 ml trifluoromethane-sulphonic acid trimethylsilyl ester are 
added. The mixture is stirred for 24 hours at room temperature, then 
filtered and the residue is washed well with methylene chloride. 
The filtrate is concentrated at 40.degree. C. to give an orange red 
resinous product. This product is chromatographed over silica gel. Elution 
is effected with ethyl acetate. On concentration of the relevant fractions 
the heading compound is obtained with a purity of 97% (HPLC). 
2) Building up of the protected octa-peptide 
17.2 g aminomethylated polystyrene (Brand Dow 0.7% by weight of N 
corresponding to 0.5 mmol amino-methyl groups per g resin) are suspended 
in 80 ml methylene chloride/DMF 4:1. Successively there are added 4.17 g 
of the end product of step 1, 1.6 g HOBT and 4.0 g DCCI. After the mixture 
is stirred for 2 hours at room temperature, the Kaiser test is negative. 
The mixture is filtered and washed. 
The washed resin is suspended in 100. ml tetrahydrofuran and methanol 3:1 
and treated with portions of 10.4 g sodium borohydride. The mixture is 
stirred for 6 hours at room temperature, filtered and the resin washed. 
The resin is suspended in methylene chloride/DMF 4.1. 5.5 g Fmoc-Cys 
(S-t-Bu)OH, 1.74 g HOBT and 3.6 g DCCI are added. The Fmoc protecting 
group is split off with piperidine (2.times.20 minutes contact time). 
In analogous manner in successive cycles the following N-Fmoc protected 
amino acids are coupled using HOBT/DCCI-ThrOH; Lys (BOC)-OH: D-TrPOH, 
Phe-OH, Cys(S-tBtu)OH and D-Phe-OH to-give the Fmoc protected octapeptide 
resin. Final loading 0.26 mmol/g. 
3) Oxidation and splitting off 
The resultant resin is suspended in 100 ml trifluoroethanol/methylene 
chloride 1:1 and treated with 50 ml tributylphosphine. The mixture is 
stirred for 70 hours at room temperature. The mixture is filtered, washed 
and treated with a 100 ml 1:1 mixture of tetrahydrofuran and 1N 
aminoacetate solution. 1.1 ml of 30% aqueous hydrogen peroxide are added. 
The mixture is stirred for 24 hours at room temperature. The resin is 
washed. The mixture is treated with 20 ml trifluoroacetic acid, 80 ml 
methylene chloride, 10 ml water, and ml thioanisole. The mixture is 
stirred for 2 hours, then filtered and washed with trifluoroacetic acid 
and methylene chloride. 200 ml diethyl ether are added to the filtrate. 
The resultant precipitate is filtered off. The residue is dissolved in 
aqueous buffer and demineralised, e.g. using Duolite. The solution is 
freeze-dried as the acetate to give the title compound as the acetate. All 
the above examples, e.g. the compounds of examples 1 and 2 may be produced 
in analogous manner. 
EXAMPLE S2 
Production of N.sup..alpha. -[.alpha.-glucosyl(1-4)-deoxyfructosyl)-SMS 
(see Example 2) 
393 g of the octapeptide bonded to the resin are produced according to the 
above example S2. The cysteine protecting groups are removed reductively. 
The peptide bond to the resin is oxidized to the cyclic octapeptide by 
hydrogen peroxide in a mixture of tetrahydrofuran/water. 
After washing in tetrahydrofuran and then DMF the peptide resin is shaken 
in 3600 ml of a mixture of DMF/ACOH.(8:1) The suspension is treated with 
526 g D(+)maltose monohydrate. The mixture is warmed to 60.degree. and 
stirred for 18 hours at this temperature. 
The mixture is cooled and the peptide resin filtered off, and successively 
washed with DMF and methanol, Then it is washed with methylene chloride. 
The peptide is then split over 1 hour from the resin with a mixture of 
2900 ml methylene chloride and 716 ml trifluoroacetic acid with a trace of 
water. 
The filtrate is then stirred and treated with portions of 597 g sodium 
carbonate, stirred for 30 minutes and filtered. The residue is washed with 
methylene chloride and methanol. 
The filtrate is concentrated to dryness. 
It is demineralised using an unfunctionalized polystyrene column like 
Duolite, or reversed phase HPLC material such as silica gel treated with 
silicone and bearing long chain fatty alcohol groups (e.g. Labomatic, 
Switzerland, Brand HB-SIL-18-20-100). The pure title compound is obtained. 
The compounds of the invention exhibit pharmacological activity and are 
therefore indicated for use as pharmaceuticals for therapy. 
The activity of the compounds of the invention may be observed in standard 
pharmaceutical and biopharmaceutical tests. The compounds are in general 
at least as potent as the unmodified peptide (i.e. the corresponding sugar 
free peptide) on administration by injection or orally. They are in 
general better absorbed, are more easily soluble in water, and have a 
longer duration of action. 
The compounds of the invention are therefore useful in the same indications 
as for the unmodified peptides. 
The compounds of the invention may be compared with the unmodified peptides 
in standard bioavailability tests. 
The compounds of the invention, for example, may be detected in the blood 
plasma for a longer period after administration than the unmodified 
peptides, as indicated in standard bioavailability experiments. 
The compounds of the invention and the unmodified peptide may be 
administered to for example dogs in a single dose sufficient to produce a 
therapeutic effect by oral or intravenous administration. 
Doses used are those which permit the peptide or a metabolite thereof to be 
detected in the blood. Detection may be effected in conventional manner, 
e.g. by radioimmunoassay. 
In the above mentioned test, it has for example been determined that the 
example 2 compound produced on oral administration a ten fold higher blood 
concentration compared with octreotide. 
The absolute bioavalability of orally and intravenously administered 
example 2 compound, measured on the basis of the AUC (area under the 
curve) is 5 times higher than that of octreotide. The elimination 
half-life on intravenous administration is about 2,3 hours compared with 
about 0.5 hour for octreotide. 
Additionally the compounds of the invention advantageously are eliminated 
to a greater extent through the kidneys. This may be observed in standard 
tests. 
Fasted Male rats (225-375 g) are administered orally with water (50 ml/kg). 
After 30 minutes the animals are anaesthetized with e.g. Inactin (100 
mg/kg i.p.). The bile duct and bladder are cannulated. Both V. jugularis 
are exposed. In one vein an infusion of glucose 5% with ethanol 1% is 
administered (5 ml/hr) to stimulate diuresis. The other vein is used to 
take blood samples (0.5 ml) every hour over 4 hours. 
The compound of the invention and the unmodified peptide is administered 
s.c. at a dose of from about 10 to about 1000 microgram/Kg. The 
concentration of the compound is determined in conventional manner e.g. by 
RIA. 
In the above test for example the following results have been obtained with 
the example 2 compound and octreotide at a dose of 10 microgram/kg: 
______________________________________ 
Percentage 
eliminated 
through 
Bile Urine 
______________________________________ 
Example 2 compound 1.6 36 
Octreotide 22 19 
______________________________________ 
Whereas octreotide is eliminated in both the bile and urine the example 2 
compound is predominantly eliminated in the urine. 
Improved absorption on oral administration may be detected for the 
compounds of the invention as follows: 
The compound of the invention and the unmodified analogue are administered 
orally to OFA rats (e.g. 10 mg/kg). After definite periods of time, e.g. 
15, 30 and 60 minutes, blood samples are collected, These are analysed for 
their drug content by e.g. RIA. 
It has for example been determined in this test that the compound of 
example 44 at a dose of 10 mg/kg exhibits a 50 to 100 per cent higher 
absorption than the unmodified peptide, oxytocin. Results are as follows: 
TABLE 
______________________________________ 
Rat plasma levels following oral 
administration. Results given in ng/ml 
15 mins. 30 mins. 60 mins. 
______________________________________ 
oxytocin 7.53 3.60 2.55 
compound of example 44 
11.79 6.98 3.98 
______________________________________ 
The pharmacological activities of the compounds of the invention may be 
investigated in standard pharmacological tests, e.g. after injection, aid, 
if desired, compared with those of the unmodified peptides, e.g. in terms 
of potency and duration of action. 
For example pharmacological tests may be effected to examine the effects of 
the compounds of the invention on hormones in animals. Thus the compounds 
which inhibit the secretion of hormones may be tested by measuring the 
lowering of blood levels of the hormone. 
Compounds of the invention which inhibit GH (growth hormone) secretion, 
especially the compounds of formula VIII, and more especially compounds of 
formula VIII a to f, and reduce the GH concentrations in the blood, may be 
tested as follows: 
Fasted rhesus monkeys (at least 5 monkeys) in primate chairs receive the 
compound of the invention in a piece of banana as vehicle. The compounds 
are administered at a dose of from about 0.1 mg/kg to about 10 mg/kg p.o. 
Blood is taken from the V. Saphena via a catheter. The GH concentration in 
the blood is measured by RIA (radio immunoassay). 
In this test with rhesus monkeys it has for example been determined that 
the example 2 compound at a dose of 0.1 mg/kg lowered the GH secretion by 
at least 50 per cent for longer than 10 hours, compared with a 5 hour 
duration of lowering effect with-the unchanged peptide, octreotide. 
A further test is as follows: 
Male rats are decapitated and blood is collected 1 hour after administering 
the GH secrection inhibiting compound in several logarithmically spaced 
doses. The GH level in the serum is determined means of RIA. In this test, 
these compounds of the invention are active at doses from about 0.02 to 
about 30 microgram/kg s.c. 
In this test it has for example beer determined that the example 1,2,21 and 
24 compounds have an ID.sub.50 of 0.045, 0.190, 0.3 and 0.2 microgram/kg 
s.c. respectively compared with the ID.sub.50 for natural stomatostatin in 
the same test of 93 microgram/kg s.c. (the ID.sub.50 indicates the amount 
of compound required to lower the GH content by 50% compared with that of 
untreated control animals). 
Unlike natural somatostin, the GH secretion inhibiting compounds of the 
invention are highly active in this test for a long period of time (e.g. 6 
hours). 
The GH-reducing activity of these compounds is also observed after oral 
application to male rats having oestradiol implants. In this test there 
are relatively small variations in the GH level, The test is carried out 
as follows: 
A loop (length 50 mm .0. 3 mm) of silastic with 50 mg of oestradiol is 
implanted under ether anaesthesis under the dorsal skin of male rats which 
have a weight of ca. 300 g. At various times (1 to 6 months later), these 
animals are used repeatedly for tests. The test substance is administered 
either s.c. or orally. 
Directly before, as well as at various times after administration of the 
substance, ca. 0.8 ml of blood is removed from the retro-orbital plexus. 
It is centrifuged and the GH level in the serum is determined by RIA. 
The compounds of the invention are, after oral administration, more active 
than the corresponding unmodified peptides, even after several hours. The 
ID.sub.50 for each of the compounds of examples 1 and 2 after two hours is 
ca. 17 to 40 times lower than that of the unmodified peptide octreotide. 
Further results are as follows: 
______________________________________ 
Compound of example 
ID.sub.50 p.o. microgram/kg 
______________________________________ 
21 500 
24 25 
Octreotide 1400 
______________________________________ 
These compounds of the invention are useful for indications where an 
inhibition of GH secretion is desired. Indications include diabetes 
mellitus, the prevention and treatment of angiopathy and proliferative 
retinopathy, as well as acromegaly. 
The GH secretion inhibiting compounds of the invention also inhibit 
pancreatic secretion. 
This inhibition may be detected in tests on animals. 
The method is described in Scand. J. Gastroint. 6, 423 (1975) by S. J. 
Konturek et al. may be used. 
The GH secretion inhibiting compounds of the invention also inhibit gastric 
acid secretion and increase the pH of the stomach juices to higher pH 
units. 
The activity of these compounds is observed in e.g. the following test: 
GH secretion inhibiting compounds of the invention are administered to 
fasted rats with fistula implanted in their stomach in doses from about 
0.05 mg/kg to about 5 mg/kg by stomach tune. After 1 hour the fistula is 
opened. The stomach Juice is collected in 30 minute periods. The collected 
volumes are registered and the acid concentration determined. 
In the above mentioned test the compound of example 2 increased the pH to 
6-8 for 3,5 hours. Octreotide increased the pH units to 6-7 for only 2 
hours. The compound of example 2 is at least 60 times more active than 
cimetidine in this test system. 
The GH secretion inhibiting compounds of the invention, especially the 
compounds of formula VIII are therefore useful in the treatment of 
gastro-intestinal disorders, e.g. in the treatment of peptic ulcers, 
gastro-intestinal bleeding, acute pancreatitis and gastro-enteropancreatic 
tumours (e.g. vipomas, insulinomas, glucagonomas, etc.). 
The GH secretion inhibiting compounds of the invention also inhibit the 
proliferation and/or keratinisation of epidermal cells, and are therefore 
useful in the treatment of dermatological illnesses which are associated 
with pathological proliferation and/or keratinisation of epidermal cells, 
especially in the treatment of psoriasis. 
Furthermore, these compounds are useful in the treatment of degenerative 
senile dementia, also the Alzheimer type (SDAT) of senile dementia, or in 
the treatment of cluster headaches. (repeated headaches). 
For these indications, the appropriate dosage will, of course, vary 
depending upon, for example, the compound employed, the host, the mode of 
administration and the nature and severity of the condition being treated. 
However, in general, satisfactory results in animals are indicated to be 
obtained at daily dosages from about 0.005, e.g. 0.03, microgram/kg to 
about 300 microgram/kg animal body weight. In larger mammals, for example 
humans, an indicated daily dosage is in the range from about 2 micrograms 
to about 2 mg of a compound conveniently administered, for example, in 
divided doses up to four times a day. 
The sugar derivatives of calcitonin and of calcitonin analogues or 
derivatives according to the invention, more especially the derivatives of 
formula X reduce the calcium plasma level. Moreover, they are functional 
antagonists of the parathormone, and effect a positive calcium balance in 
the bones. The hypocalcemic activity of the new compounds may be measured 
in known manner, e.g. according to the method of M. Azria et al., reported 
in the Calcitonin 1984 Symposium, 24th October, Milan and published as 
"Short Communication" in "Current Clinical Practice Series" No. 42, 
Excerpta Medica 1986, page 104. In this method, a Ca.sup.2+ -ion selective 
electrode is used, so that the content of calcium ions in the blood of 
young rabbits or dogs may be continuously measured. The compounds are 
administered i.v. at a dosage of from about 0.1 to about 10 micrograms/kg, 
e.g. conforming to ca. 1 international unit per kg. The measurements are 
carried out over 5 hours and the AUC "area under the curve" is calculated. 
The compounds can also be tested in other tests, e.g. in the hypocalcemic 
standard test of M. Kumar et al., J. Endocrinology (1965), 33, page 46g., 
on rats in different dosages giving a hypocalcemic activity of 300 to 6000 
international units per mg for the hypocalcemic compounds according to the 
invention. 
It has for example been determined that each of the compounds of examples 
37 and 38 have a duration of action that is much longer than with the 
unmodified peptide, when administered i.v. to dogs (5 .mu.g/kg). In this 
test, after 3 hours a reduction in the calcium level in the blood of 15 to 
18% was observed for compounds C and D; after 6 hours, a calcium reduction 
could no longer be detected for the unmodified peptide,; whereas after 
example 37 and 38 compounds, the reduction in the calcium level was still 
as pronounced as after 3 hours. 
The hypocalcemic compounds of the invention are thus useful for all 
conditions in which a reduction of the plasma calcium level or an effect 
on bone metabolism is desired, e.g. hypercalcemia, a result of a 
deficiency in the endogenic thyrocalcitonin through a loss of thyroid 
tissue or hyperfunction of the parathyroid bland. They are also indicated 
for all bone conditions which are based on increased bone friability or in 
which a calcium fixation in the bone is desired, e.g. osteoporosis of 
various kinds (e.g. postclimacteric, posttraumatic, caused by 
corticosteroid therapy or by inactivity, malignant dieseases etc.), 
fractures, osteomalacia, rickets and renally-induced osteodystrophy, pain 
e.g. pain in the bones in connection with osteoporosis, neurodystropic 
illnesses, Paget's disease, and in combined therapy with calcium 
phosphate. 
For these calcium-related indications, the appropriate dosage will, of 
course, vary depending upon, for example, the compound employed, the host, 
the mode of administration and the nature and severity of the condition 
being treated. In larger mammals, for example humans, an indicated 
parenteral daily dosage is in the range from about 5 to about 1500 IU of a 
compound conveniently administered, for example as a single dose or if 
desired every 2 to 3 days. 
Compounds of the invention, which are sugar derivatives of LHRH or 
analogues thereof, inhibit lutenizing hormone secretion, e.g. as indicated 
by an inhibition of ovulation in animals. 
This test is effected according to M. Marko and E. Fluckiger, Experientia 
30, 1174-1176 (1974): 
Adult female rats of the Ivanovas Wistar strain (Sprague Dawley, Iva:SDIV, 
200-250 g) are kept under standard conditions: 14 h light (from 04.00 to 
18.00 hours); 24.degree. C.; 55-60% rel.humidity; food and water ad 
libitum. 
Animals with regular 4-day cycles are injected on proestrus day at 13.00 h 
with the compound, subcutaneously or by the oral route. The next day at 
9.00 a.m. the rats are sacrified and ova counted on both Fallopian tubes 
with the aid of a dissecting microsope. Only when no eggs are found is 
ovulation considered to be inhibited. The mean number of eggs per 
ovulating rat in each treatment groups is also determined. 
In general these compounds of the invention are effective in a range from 
about 0.0005 to about 10 mg/kg. For example the example 45 compound is 
active at 0.01 mg/kg s.c. The inhibiting effect on luteinizing hormone 
secretion of the compound can also be tested in vitro: Pituitary cell 
cultures are prepared according to the method of Vale (W. Vale and G. 
Grant: Methods in Enzymology 37, 82-93 (1975) as has been described 
previously (M. Marko and D. Romer: Life Sciences, 33, 233-240 (1983). 
Primary cultures are maintained for 4 days in an incubator at 37.degree. 
C. Thereafter the cells are washed and incubated for 3 hours in 1 ml 
medium containing LHRH or the test compound. At the end of the incubation, 
the supernatant is removed and assayed for LH by specific 
radioimmunoassay. 
In this test in general the test compounds are found to be effective in a 
range from about 10.sup.-12 to about 10.sup.-7 M concentration, inhibiting 
the LHRH-induced LH secretion in a dose-dependent manner. 
For these LHRH indications, the appropriate dosage will, of course, vary 
depending upon, for example, the compound employed, the host, the mode of 
administration and the nature and severity of the condition being treated. 
However, in general, satisfactory results in animals are indicated to be 
obtained at daily dosages from about 0.005 microgram/kg animal body 
weight. In larger mammals, for example humans, an indicated daily dosage 
is in the range from about 2 mg to about 20 mg of a compound conveniently 
administered, for example, in divided dosed up to four times a day. 
For the example 2 compound-an indicated dose is from 3 to 10 mg three times 
a day, e.g. p.o. for diabetes or ulcers. 
The compounds of the invention may be administered by any conventional 
route, for example enterally, e.g. orally, e.g. in the form of drinking 
solutions, tablets or capsules, nasally, e.g. in the form of liquid or 
powder sprays and ointments., or parenterally, e.g. in the form of 
injectable solutions or suspensions. 
The appreciate dosage of the compounds of the invention for any particular 
route of administration, e.g. by nasal, or oral, may be ascertained by 
standard bioavailability trials using the same substance injected i.v., 
i.m., or s.c. In general for oral administration the daily doses are about 
10 to about 100 times higher than that available for injection i.m. or 
s.c. 
The compounds of the invention may be administered in any pharmaceutically 
acceptable form, e.g. in free form, e.g. free base form or, when the 
compound is an acid, in free acid form, or in pharmaceutically acceptable 
salt form. The salt form may be for example an acid addition salt form or 
when the compound is an acid in pharmaceutically acceptable cationic salt 
form. The compounds may also be administered in complex form. The 
compounds may additionally or alternatively be in the form of a solvate, 
e.g. a hydrate. 
The compounds of the invention exhibit the same order of activity in each 
of these form. 
The present invention also provides pharmaceutical compositions for a 
compound of the invention in pharmaceutically acceptable form in 
association with at least one pharmaceutical carrier or diluent. Such 
compositions may be manufactured in conventional manner. 
Unit dosage forms of the invention contain 0.5 micrograms to 10 mg of the 
compounds, e.g. for the GH and LH indications. 
A drink ampoule or injectable solution may contain per ml for example 8.5 
mg of the example 2 compound in acetate form, 11.45 mg citric acid, 6.32 
mg NaOH, 4.5 mg NaCl. 
The present invention also provides a compound of the invention for use in 
any indication mentioned above, including lowering GH secretion, diabetes 
mellitus, reducing gastric secretions and acromegaly for the somatostatin 
like compounds of the invention. 
The present invention also provides the use of a compound of the invention 
in the manufacture of a medicament suitable for use in the treatment of 
any indication mentioned above, including lowering GH secretion, diabetes 
mellitus, reducing gastric secretions and acromegaly for the somatostatin 
like compounds of the invention.