FR2619566A1 - Process for preparing a peptide-alcohol - Google Patents

Abstract

The invention relates to a process for preparing a peptide-alcohol which comprises at the C-terminal end of the peptide chain 2 alcohol groups or one alcohol group and one thiol group, characterised in that the peptide-alcohol is prepared by acid hydrolysis of an acetal of the peptide-alcohol and of a resin containing formyl-phenyl residues.

Description

 The present invention relates to novel peptide derivatives, their preparation and their use as medicaments.

 In particular, the invention relates to a glucosylated derivative of a biologically active peptide, which derivative has a prolonged duration of action relative to the unmodified peptide and contains at least one of the amino acid residues, a carbohydrate residue attached to the amino group of the amino acid residue by a bond other than a direct N-glucosidic bond and, when the derivative is a condensation product of a carbohydrate containing a carboxy group with a peptide having less than 8 amino residues -acids, by a bond other than an amide bond.

 These compounds will be referred to hereafter as the compounds of the invention.

By unmodified peptide is meant the peptide of corresponding structure having no carbohydrate residue
The Applicant has further found that the compounds of the invention are indicated by pharmacological properties particularly interesting and surprising, especially by a longer duration of action, for example as described below.

It has been found that the incorporation of one or more carbohydrate moieties even when bonded differently than normal glucosidation eg of Asn or Ser results in such properties.

 These carbohydrate moieties are preferably introduced to amino groups of amino acids remote from the active site of the peptide.

 The term peptide used in the present application includes peptides, for example di- and tri-peptides, oligopeptides, polypeptides and proteins. Preferably, the peptide contains more than 7 amino acid residues. Advantageously, the peptide contains from 8 to 32 amino acid residues.

By amino acid residue is also meant an amino alcohol residue corresponding to the amino acid residues.

 The term biologically active peptide used in the present application covers, in particular, compounds having a pharmacological or therapeutic activity, for example compounds having hormonal, enzymatic or immunomodulatory activity, or which stimulate or inhibit such activity. These biologically active peptides include natural peptides isolated from sourenengetal or by cell fermentation, for example by genitally engineered or synthetically derived, and also their derivatives or the like.

 By derivatives and analogs of these peptides is meant in particular the natural peptides in which one or more amino acid residues have been deleted and / or substituted by one or more other amino acid residues and / or in which one or more functional groups have been replaced by one or more other functional groups and / or in which one or more groups have been replaced by one or more other isosteric groups. In general, the term covers all modified derivatives of a biologically active peptide that exhibit a qualitatively similar effect to that of the unmodified peptide.

 The carbohydrate used can be, for example, any known mono-, di-oligo-saccharide, especially a mono-, di- or trioseose or a derivative thereof, for example one of its amine derivatives and and / or carboxyfe and / or reduced and / or esterified.

 The carbohydrate may be coupled, for example, to an N-terminal amino group and / or to at least one amino group of the peptide present in the side chain of this peptide.

 The carbohydrate can be coupled by one of its functional groups to the peptide either directly or indirectly via a bridge, for example an alkylenecarbonyl group. This coupling can be carried out according to the known methods, especially as described below.

 Preferred compounds of the invention are those wherein the carbohydrate moiety is attached to an amino group of the peptide by a linkage other than a direct N-glucoside linkage or an amide linkage.

 A group of compounds of the invention may be prepared by rearranging Amadori or Heyns.

 The invention also includes orally administrable pharmaceutical compositions containing a compound of the invention, especially those having at least 8 amino acid residues.

dans laquelle

représente le reste désoxy d'un cétose, le groupe
CH2 de ce reste étant lié au groupe NH d'un peptide biologiquement actif, et
P représente le reste d'un peptide biologiquement actif de formule NH2-P, dans laquelle le groupe
NH2 représente le groupe amino N-terminal du peptide P ou un groupe amino situé dans une channe latérale du peptide P, b) de formule II

dans laquelle

The present invention relates in particular to the following glucosyl derivatives of biologically active peptides a) of formula I

in which

represents the deoxy residue of a ketosis, the group
CH2 of this residue being linked to the NH group of a biologically active peptide, and
P represents the residue of a biologically active peptide of formula NH 2 -P, in which the group
NH 2 represents the N-terminal amino group of the peptide P or an amino group located in a side channel of the peptide P, b) of formula II

in which

dans lesquelles
P represente le reste d'un peptide biologiquement
actif de formule NH2-P, les restes

representent des restes glucidiques,
R représente l'hydrogène ou un groupe alkyle
en C1-C3 ou alcanoyle en C1-C4 et
Q, Q', Q" et Q"' représentent des groupes couplant
le reste du peptide avec le reste du glucide,
le groupe NH lié à P représentant le groupe amino N-ter
minal du peptide P ou un groupe amino situé dans la channe
latérale du peptide P, ou e) de formule Va ou Vb HOH2C-(CHOH)C-CY-CH2-NH-P (Va)

represents the deoxy residue of an aldose, the binding
free from this residue being linked to the NH group of a peptide
biologically active, and
P represents the rest of a biologically peptide
active ingredient of formula NH2-P, in which the group
NH 2 represents the N-terminal amino group of the peptide P or a
amino group located in a lateral channe of the pep
P, c) of formula III G3-CO-NR-P (III)
in which
G3CO represents the rest of a uronic acid or
a polyhydroxymono- or dicarboxylic acid,
R is hydrogen or an alkyl group
C1-C3 or C1-C4 alkanoyl, and
P represents the rest of a biologically peptide
active ingredient of formula NH2-P, containing at least
8 amino acid residues,
NRy being located on the N-terminal amino group of the peptide P
or in a side chain of peptide P, d) of formula IVa, IVb, IVc or IVd

in which
P represents the rest of a peptide biologically
NH2-P formula, the residues

represent carbohydrate residues,
R represents hydrogen or an alkyl group
C1-C3 or C1-C4 alkanoyl and
Q, Q ', Q "and Q"' represent coupling groups
the rest of the peptide with the rest of the carbohydrate,
the NH group linked to P representing the amino group N-ter
of the peptide P or an amino group located in the channe
peptide P, or e) of formula Va or Vb HOH2C- (CHOH) C-CY-CH2-NH-P (Va)

in which
Y represents H2 or H, OH
c is 2 or 3 or 4, and
P represents a residue of a biologically peptide
active formula NH2-P,
the NH group linked to P representing the amino group
N-terminal peptide P or an amino group located in
the lateral channe of the peptide P, and any
free hydroxy groups in the rest of the polyol
of the compound of formula Va or Vb is optionally
bound by a glucosidic bond to a mono-, di- or
reducing oligosaccharide or amino sugar, as well as the acid addition salts and the complexes of these derivatives, with the conditions that a) in the compounds of formula I above, P is
other than a remainder of a gastrin peptide having
a terminal group ending in -Asp-Phe-NH2, b) when in the compounds of formula IVb Q 'means
a phenyl ring containing a divalent radical
or when in the compounds of formula IVd Q "
means the rest of an aliphatic dicarboxy acid
liquefaction, P-NH2 is other than natural insulin, c) when in the compounds of formula III G3-CO
represents the remainder of a possible muramic acid
N-acylate, the second amino acid residue to
the N-terminal end of the peptide P-NHR is other y
that the rest of an amino-dicarboxylic acid
All of the above-mentioned oligomeric residues may be mono-, di- or oligosaccharides. These carbohydrates may contain heptoses, hexoses and / or pentoses which may exist in pyranic or furan form.

In all formulas I to V shown above, only one carbohydrate residue is represented by peptide residue. However, the invention also covers glucosylated derivatives of peptides having more than one free amino group on the peptide residue, these derivatives containing 2 or 3 carbohydrate residues per peptide residue.
The invention further relates to all biologically active peptides which have more than one bonded carbohydrate moiety as previously indicated.

 The glucosyl peptides preferably contain 1 to 3 monosaccharide residues, which can be linked together to form a disaccharide or tri-saccharide.

 In all the compounds mentioned above, the symbol rJ indicates that the connection may be in position a or B.

représente de préférence a) un reste de formule VIa

In formula I, the rest

preferably represents a) a radical of formula VIa

in which one of the symbols G a and Gb means hydrogen and
the other OH, one of the symbols Gc and Gd means hydrogen and
the other OH or a glucosyloxy residue in which
the glucosyl residue can be derived from a mono-, di
or reducing oligosaccharide,
one of the symbols G and Gf represents hydrogen
e
and the other oh,
one of the symbols G and Gh represents hydrogen
and the other OH or CH20H,
for example in which the symbols Ga to Gh
are chosen in such a way that the rest of
formula VIa corresponds to a remainder that can be
obtained by rearranging Amadori from a
mono-, di- or oligosaccharide natural or accessible
by synthesis.

Examples of carbohydrate radicals of formula VIa are deoxyfructosyl, deoxytagatosyl, deoxysorbosyl, α-glucosyl- (1-4) -deoxyfructosyl, α-glucosyl (1-4) -α-glucosyl (1-4) residues. - deoxyfructosyl b) A remainder of formula VIb

in which
one of the symbols G a and G b means hydrogen and
the other oh,
one of the symbols G c and Gd means hydrogen and
the other OH or glucosyloxy in which the rest
glucosyl can be derived from a mono-, di- or oligo
saccharide reducer,
one of the symbols G e and G f means hydrogen and
the other means hydrogen, COOH, CH20H,
CH 2 -O-P (O) - (OH) 2 or CH 2 O -glucosyl, wherein the
glucosyl residue can be derived from a mono-, di- or
reducing oligosaccharide, for example wherein the symbols G a to Gf are chosen such that the radical of formula VIb corresponds to a residue which can be obtained by rearranging Amadori from a natural mono-, di- or oligosaccharide or accessible by synthesis.

 The radicals of formula VIb can be obtained for example by rearranging Amadori from saccharides such as gentiobiose, melibiose, ribose, xylose or from uronic acids such as glucuronic acid or galacturonic acid.

signifie de préférence a) un reste de formule VIIa

In formula II, the remainder of formula

preferably means a) a remainder of formula VIIa

in which
one of the symbols Ga and Gb means hydrogen
and the other a direct link,
one of the symbols G c and Gd means hydrogen and
the other oh,
one of the symbols G e and Gf means hydrogen
and the other OH or glucosyloxy in which the rest
glucosyl can be derived from a mono-, di- or oligo
saccharide reducer,
one of the symbols Gg and Gh means hydrogen and
the other CH2OH or CH2-O-glucosyl, in which the
glucosyl residue can derive from a mono-, di- or
reducing oligosaccharide, for example wherein the symbols G a to G h are chosen such that the remainder of formula VIIa corresponds to a residue which can be obtained by rearranging Heyns from a natural mono-, di- or oligoketose or accessible by synthesis, b) a remainder of formula VIIb

in which
one of the symbols G a and G b means hydrogen
and the other a direct link,
one of the symbols G c and Gd means hydrogen
and the other oh,
one of the symbols G e and G f means hydrogen and
the other CH 2 OH or CH 2 -O-glucosyl, wherein
the glucosyl residue can be derived from a mono-, di
or reducing oligosaccharide,
for example in which the symbols Ga to Gf
are chosen in such a way that the rest of
formula VIIb corresponds to a remainder that can be
obtained by Heyns rearrangement from a
mono-, di- or oligoketosis natural or accessible
by synthesis.

 The radicals of formula VIIa or VIIb can be obtained for example by Heyns rearrangement of a carbohydrate such as D-fructose, lactulose, L-sorbose, D-tagatose or D-ribulose.

 In formula III, the polyhydroxymono- or dicarboxylic acid, for example, contains at least 3 hydroxy groups and may also contain other substituents, for example amino or acetylamino groups.

 Examples of such polyhydroxycarboxylic acids include onic acids derived from carbohydrates such as gluconic acid, or aric acids such as glucaric acid, as well as quinic acid, acetylmuramic acid and acetylneuraminic acid. and D-glucosaminic acid.

 Examples of uronic acids include glucuronic acid and galacturonic acid.

 In the compounds of formula IV, G4, G4, G4 and G4 'have the meanings given above for G1 or G2.

 The Q or Q 'symbol connects an NH 2 group of the peptide with an NH 2 or OH group of the carbohydrate residue and means, for example, the remainder of a dicarboxylic acid or preferably a -CbH 2 -CO residue in which b is from 0 to 6. The rest can be branched.

ou en particulier un reste -CbH2b-CO- (b = par exemple de 1 à 6). Q signifie par exemple un reste -CH2-CO-.Q 'means for example a remainder

or in particular a residue -CbH2b-CO- (b = for example from 1 to 6). Q means for example a residue -CH2-CO-.

Q means especially -CO- or -CS-. -NH-Q "- and -NH-Q"'- mean a residue which connects the NH2 group of the peptide with the carbohydrate residue, especially the residues of ß-aminocarboxylic acids. They can mean, for example, a residue -NH-CbH2b-CO
Preferred compounds of formula V are those of formula Va, especially those wherein n is 3.

 All the P residues mentioned above are residues of biologically active peptides.

Such peptides comprise all the natural or synthetic peptides as well as, as indicated at the beginning of the present application, the derivatives and analogs of these peptides having a hormonal, enzymatic or immunomodulatory activity. This activity can be both stimulating and inhibitory. By way of examples, mention may be made of the following peptides: somatostatin, calcitonin, oxytocin, vasopressin, insulin, LH (luteinizing hormone), LHRH (gonadorelin) GRF (Factor triggering the secretion of somatotropic hormone), gastrin, substance P, cathepsin, enkephalins, as well as all their derivatives and analogues having an activity similar to that of the peptide or having an antagonistic activity.

 The compounds of the invention preferably contain at least 8 amino acid residues, for example from 8 to 32, especially from 8 to 20, in particular from 8 to 10, amino acid residues.

 The preferred peptides are those corresponding to formulas I and II.

 For the sake of simplicity, in the preceding and following formulas the carbohydrate residue is represented in pyran form. It goes without saying that the invention also includes furan forms and linear forms of these carbohydrates, as far as they exist for the carbohydrates concerned.

 The invention also relates to a process for preparing the compounds of the invention. These compounds can be prepared according to generally known methods for the synthesis of compounds of this type.

The compounds of the invention may be prepared for example as follows: a) at least one protecting group present is removed
in a glucosylated peptide of the protected invention, or b) two peptide units are coupled by an amide linkage.
diques, each of which contains at least one amino
acid or an amino alcohol in protected form or
unprotected and of which a peptide unit contains
the carbohydrate residue, the coupling being carried out
in such a way that the amino sequence is obtained
desired acids, and optionally implemented
step a) of the process, or c) introducing at least one optionally carbohydrate residue
protected in a protected or unprotected peptide and possibly
step a) of the process is carried out, or d) a functional group of a peptide is converted into
glucosylated protected or unprotected invention,
into another functional group or one eliminates
this functional group, and possibly
step a) of the process is carried out, or e) a glucosylated peptide of the invention is oxidized in which
The mercapto group of Cys remains are in free form,
to produce a peptide in which the
two remains Cys are connected by a bridge -SS.

 As indicated at the beginning of this description, the term "carbohydrate" as used in the present application also covers carbohydrate derivatives such as amino oses, oxidized and reduced oses or esterified oses.

 The above reactions can be carried out analogously to the process described in the following examples, in particular processes a) and b) can be carried out according to the synthesis of the invention described hereinafter. If desired, protecting groups that are suitable for use in peptides or carbohydrates can be used in these reactions to protect functional groups that do not participate in the reaction. The term protecting group includes a polymeric resin having functional groups.

 The compounds of formula I may be prepared by reacting, in a slightly acidic medium, a protected peptide having a free amino group with a corresponding reducing mono-, di- or oligosaccharide or uronic acid or an ester of this acid (Amadori rearrangement), and then removing the protecting groups.

 This reaction can be carried out conventionally for a rearrangement of Amadori.

The added acid may be, for example, glacial acetic acid. When reacted with a uronic acid, it is not necessary to add another acid.

An excess of carbohydrate is preferably used, for example equivalent for one peptide equivalent.

The reaction may be carried out in a polar solvent such as methanol, preferably at temperatures of about 60 to 700 ° C.

 The compounds of formula II can be prepared by reacting in a weakly acidic medium a protected peptide having a free amino group with a ketosis (Heyns rearrangement). The reaction can be carried out under the same conditions as a rearrangement of Amadori, as indicated above.

 Compounds of formula III may be prepared by reacting a protected peptide having a free amino group with an acid of formula G3-COOH or a reactive derivative thereof, and then removing all protecting groups. This reaction can be a conventional amidation which can be carried out in a manner known per se. As reactive derivatives of the carboxylic acid, it is possible in particular to use the acid halide. The amides can also be prepared, for example, by reacting the free acid in the presence of hydroxybenzotriazole and dicyclohexylcarbodiimide.

The compounds of formula IVa, IVb, IVc and
IVd can be prepared by a) first reacting the peptide with the compound
bridging then the product obtained with the carbohydrate,
or b) by first reacting the carbohydrate with the compound
bridging then the glucosylated bridge compound
thus obtained with the peptide.

 These reactions can be carried out according to known methods. The compounds formed are mainly amides, esters or acetals. The compounds of the invention can be purified according to known methods.

dans laquelle
L signifie 0 ou S et
G4 a la signification donnée précédemment, et dans laquelle les groupes hydroxy libres présents dans G4 sont sous forme protegée, par exemple par acylation, avec un peptide P-NH2 sous forme protégée, et on elimine ensuite les groupes protecteurs.The compounds of formula IVa in which
Q is -CO- or -CS- may be prepared, for example, by coupling the corresponding isocyanate or glucosyl isothiocyanate of the formula

in which
L means 0 or S and
G4 has the meaning given above, and wherein the free hydroxy groups present in G4 are in protected form, for example by acylation, with a peptide P-NH2 in protected form, and then the protecting groups are removed.

 The reaction can be carried out according to known methods for the preparation of urea derivatives.

 The compounds of formula IVc and IVd can be obtained for example by rearranging Amadori or Heyns, for example as described above for the preparation of the compounds of formulas I and II.

The compounds of formula Va or Vb can be prepared for example a ') by reductive amination of an aldose, a deoxy
aldose or ketosis with the peptide P-NH2, or b ') by reduction of the hemi-acetal group in a compound
of formula I or II, each product participating in the reaction may, if desired, be provisionally protected.

 The reducing nation friend and the reduction can be made according to the known methods.

The reductive nation friend can be carried out for example with NaBH 3 CN, preferably at pH 7. The reduction of the hemi-acetal group can be carried out with borohydride, for example with NaBH 4, preferably at a pH of about 6.

 Starting materials used in the foregoing reactions are known or can be prepared according to known methods.

dans laquelle
A représente l'hydrogène, ou un groupe alkyle en C1 -C3
ou alcanoyle en C1-C4, > N-CH(Zl)-CO signifie
1) un reste (L)- ou (D)-phénylalanine éventuellement
substitué par des halogènes ou par des groupes
N02, NH2, OH,alkyle en C1-C3 et/ou alcoxy en
C1 C3, ou
2) le reste d'un a-amino-acide lipophile naturel
ou d'un (D)-amino-acide correspondant autre
que celui indiqué sous 1),
Z1 dans le reste )N-CH(Z1)-CO- represente le reste
d'un amino-acide défini sous 1) et 2),
A' signifie l'hydrogène ou un groupe alkyle en C1-C3,
Y1 et Y2 signifient,indépendamment l'un de l'autre, 1) l'hydrogène

où m signifie un nombre entier de 1 à 4 inclus,
Ra signifie CH3 ou C2H5 et R b signifie H, CH3 ou C2H5,

A preferred class of glucosylated peptides comprises the glucosyl derivatives of somatostatin, for example containing from 4 to 9 amino acids. The term somatostatin includes somatostatin analogs and derivatives thereof. Particularly preferred glucosyl derivatives are the glucosyl derivatives of peptides of formula VIII

in which
A represents hydrogen, or a C1-C3 alkyl group
or C 1 -C 4 alkanoyl,> N-CH (ZI) -CO means
1) a residue (L) - or (D) -phenylalanine optionally
substituted by halogens or by groups
NO 2, NH 2, OH, C 1 -C 3 alkyl and / or alkoxy
C1 C3, or
2) the rest of a natural lipophilic α-amino acid
or a (D) amino acid corresponding other
than that indicated under 1),
Z1 in the rest) N-CH (Z1) -CO- represents the rest
an amino acid defined under 1) and 2),
A 'means hydrogen or a C1-C3 alkyl group,
Y1 and Y2 mean, independently of each other, 1) hydrogen

where m is an integer from 1 to 4 inclusive,
Ra means CH3 or C2H5 and Rb is H, CH3 or C2H5,

ou Rd signifie le reste d'un a-amino-acide naturel (y compris l'hydrogène) situe sur l'atome de carbone a et
Re signifie un groupe alkyle en C1-C5, ou

dans laquelle
Ra' et Rb', indépendamment l'un de l'autre, signifient
l'hydrogène , CH3 ou C2H5,
R8 et R9 , indépendamment l'un de l'autre, signifient
l'hydrogène, F, C1, Br, alkyle en C1-C3 ou alcoxy
en C1-C3, p signifie O ou 1, q signifie O ou 1, et r signifie 0, 1 ou 2, ou bien Y1 et Y2 forment ensemble une liaison,
B signifie Phe ou Phe substitué dans le cycle phényle
par F, C1, Br, N02 , NH2, OH, alkyle en C1-C3 ou
alcoxy en C1-C3,
C signifie L- ou D-Trp éventuellement substitué dans
le cycle phényle par F, C1, Br, NO2, NH2, OH,
alkyle en C1 -C3 ou alcoxy en C1-C3,
D signifie Lys, dans lequel le groupe a-amino peut
etre substitué par un groupe méthyle,
E signifieThr, Ser, Val,
F signifie COOR1, CH20R2, CO-NR3R4 ou

where n is an integer from 1 to 5 inclusive, 4) -CO-NHRc
where Rc is a linear or branched alkyl group
in C1-C6,

or Rd means the residue of a natural α-amino acid (including hydrogen) located on the α-carbon atom and
Re represents a C1-C5 alkyl group, or

in which
Ra 'and Rb', independently of one another, mean
hydrogen, CH3 or C2H5,
R8 and R9, independently of one another, mean
hydrogen, F, C1, Br, C1-C3 alkyl or alkoxy
at C1-C3, p is O or 1, q is O or 1, and r is 0, 1 or 2, or Y1 and Y2 together form a bond,
B means Phe or substituted Phe in the phenyl ring
by F, C1, Br, NO2, NH2, OH, C1-C3 alkyl or
C1-C3 alkoxy,
C means L- or D-Trp optionally substituted in
the phenyl ring by F, Cl, Br, NO2, NH2, OH,
C1-C3 alkyl or C1-C3 alkoxy,
D is Lys, in which the α-amino group can
to be substituted by a methyl group,
E means Thr, Ser, Val,
F stands for COOR1, CH20R2, CO-NR3R4 or

R1 is hydrogen or C1-C3 alkyl,
R2 means hydrogen or the remainder of a physio ester
logically acceptable and physiologically lysable,
R3 is hydrogen or C1-C3 alkyl,
phenyl, or phenylalkyl C7-C10, R3 meaning
only hydrogen or a methyl group when
R4 is -CH (R5) -X1,
R4 is hydrogen or C1-C3 alkyl, or

R5 represents the residue of a natural amino acid (including
taken hydrogen) located on the carbon atom a,
OH-CH 2 -CH 2 - or OH (-CH 2) 3, the group of
formula IX can have the configuration L or D
X1 means COOR1, CH20R2 or

R6 is hydrogen or C1-C3 alkyl,
R7 is hydrogen or C1-C3 alkyl,
or C 7 -C 10 phenylalkyl,
the residues B, D and E which can exist in L form and the residues in position 2 and 7 as well as the residues γ 14) and Y24) which can exist independently in D or L form, as well as the salts and complexes of these compounds .

 Such compounds are described in U.S. Patent No. 4,395,403, the contents of which are incorporated herein by reference.

In the polypeptide derivatives of formula
VIII above, the following definitions taken singly or in combination are preferred:
When> N-CH (Z1) -CO- has the definition given under 1), this remainder is preferably a residue of
(L) - or (D) -phenylalanine or a residue of (L) - or (D) tyrosine (where Z is benzyl or p-OH-benzyl), especially a (D) -phenylalanine residue.

 When the radical ZN-CH (Z1) -CO- has the definition given under 2), Z1 preferably denotes an alkyl group containing 3 carbon atoms, preferably 4 or more carbon atoms, for example up to 7 atoms. of carbon.

 The remainder> N-CH (Z1) -CO- is more preferably a residue as defined under 1).

 Y1 and Y2 preferably have the meanings given above under 1, 2 or 4, and most particularly together form a bond.

 B preferably means Phe or Tyr.

 C preferably means - (D) -Trp.

D means Lys
E preferably means Thr.

dans lequel le radical -CH(R5)-X1 a de préférence la configuration L.F preferably means especially

wherein the -CH (R5) -X1 radical preferably has the L configuration.

i-butyl, CH2CE2oH ou (CH2)3-OH, spécialement
CH2OH ou

en particulier

R3 preferably means hydrogen,
R5 preferably means CE120R,

i-butyl, CH2CO2OH or (CH2) 3-OH, especially
CH2OH or

in particular

ou CH20R2, spécialement CH20R2,
R2 signifie de préférence l'hydrogène.X1 Preferably means

or CH20R2, especially CH20R2,
R2 preferably means hydrogen.

 R 2, as the residue of an ester, preferably means HCO, C 2 -C 12 alkylcarbonyl, C 8 -C 12 phenylalkylcarbonyl or benzoyl.

 The remains in position 2 and 7 are preferably the L configuration.

dans laquelle Q signifie CO c ou CS
(VIIIe)

The glucosyl derivatives of somatostatin which are particularly preferred are those which have a carbohydrate residue on the N-terminal amino group, for example the compounds of formula

where Q means CO c or CS
(VIII)

 Particularly preferred compounds are the compounds of formula VIIa, VIIIb, VIIIe and VII If.

dans laquelle
AP signifie le reste désoxy d'un cétose ou d'un acide
uronique correspondant, le groupe CH2 de ce reste
étant lie au groupe NH, ledit groupe desoxy pouvant
etre obtenu par réaction d'Amadori d'un aldose
ou d'un acide uronique correspondant avec le groupe
amino libre de la somatostatine et Z1' A', Y1, B , C, D, E, Y2 ont les significations
données précédemment pour la formule VIII.A group of compounds comprises the compounds of formula VIIIpa

in which
AP stands for the deoxy residue of a ketosis or an acid
corresponding uronic, the CH2 group of this remainder
being bonded to the NH group, said desoxy group being
to be obtained by Amadori reaction of an aldose
or a corresponding uronic acid with the group
amino free from somatostatin and Z1 'A', Y1, B, C, D, E, Y2 have the meanings
previously given for formula VIII.

dans laquelle
G représente le reste acyle d'un acide uronique, d'un
acide polyhydroxycyclohexanecarboxylique, d'un acide
N-acétylmuraminique ou d'un acide N-acétyl-neurami
nique,
A représente l'hydrogène ou un groupe alkyle en C1-C3 ou
alcanoyale en C1-C4, et Z, A', Y1, B, C, D, E, Y2 et F ont les significations données précédemment.Another group of compounds comprises compounds of formula

in which
G represents the acyl residue of a uronic acid, a
polyhydroxycyclohexanecarboxylic acid, an acid
N-acetylmuraminic or N-acetyl-neuraminic acid
picnic,
A represents hydrogen or a C1-C3 alkyl group or
C1-C4 alkanoyale, and Z, A ', Y1, B, C, D, E, Y2 and F have the meanings given above.

 Advantageously, G represents the remainder of a glucuronic, galacturonic or quinic acid.

dans laquelle
Q represente l'hydrogène ou le reste glucosylique d'un
mono- , di- ou oligosaccharide, n signifie un nombre entier de 1 à 6 inclus,
A signifie l'hydrogène, un groupe alkyle en C C3
ou alcanoyle en C1-C4, et
Z , A', Y1, B, C, D, E, Y2 et F ont les significations données précédemment.Another group of compounds comprises the compounds of formula VIIIpc

in which
Q represents hydrogen or the glucosyl residue of a
mono-, di- or oligosaccharide, n means an integer from 1 to 6 inclusive,
A is hydrogen, C3-C3 alkyl
or C 1 -C 4 alkanoyl, and
Z, A ', Y1, B, C, D, E, Y2 and F have the meanings given above.

 Another preferred class of glucosyl peptides is glucosyl derivatives of calcitonin.

 The term calcitonin includes naturally occurring calcitonins (obtained from natural sources, cell cultures etc. or synthetically derived) and derivatives and the like.

 Natural calcitonins that can be used include human calcitonin, salmon, eel, chicken, beef, mutton, rat or pork, especially human calcitonin, salmon, chicken and eel.

 Derivatives and analogues of these calcitonins include in particular natural calcitonins in which one or more amino acid residues have been replaced by one or more other amino acid residues and / or the SS-bridge has been replaced by an alkylene bridge. and / or in which one or more amino acid residues have been deleted.

dans laquelle
R signifie H ou R"CO
R"CO représente le radical acyle d'un acide carboxy
lique,
Y1' représente le reste situé sur l'atome de carbone a
d'un a-amino-acide,
Y2' représente le reste situé sous l'atome de carbone a
d'un a-amino-acide, ou un reste

-CH2-S-S-CH2-CH2-COOH,
(CH2)sCOOH ou -CH2 S Y3, Y3 signifie un groupe alkyle en C1 -C4, benzyle éven
tuellement substitue par methyle ou methoxy, ou
CH3CONH-CH2-, o signifie un nombre entier de 1 à 4,
A6 signifie Thr ou D-Thr s signifie un nombre entier de 3 a 5,
A8 représente le reste amino-acyle d'un L-a-amino-acide
lipophile neutre, Ag représente le reste amino-acyle d'un L- ou D-a-amino
acide lipophile neutre, et
Z1 signifie un reste polypeptidiquesitué en position 10
à 31 d'une calcitonine naturelle ou l'un de ses
dérives ou analogues , qui possede une activité hypocal cémi ante, les 1 à 4 restes Y1 dans la formule X pouvant être identiques ou indépendamment différents , et, à l'exception du reste amino-acyle A8, tous les restes d'amino-acidesdans la formule X ont la configuration
L ou D ainsi que les sels et complexes de ces composés.Particularly preferred glucosyl derivates of calcitonins are the glucosyl derivatives of calcitonins corresponding to the formula X

in which
R means H or R "CO
R "CO represents the acyl radical of a carboxy acid
lic,
Y1 'represents the residue located on the carbon atom a
an α-amino acid,
Y2 'represents the rest under the carbon atom a
of an α-amino acid, or a residue

-CH2-SS-CH2-CH2-COOH,
(CH 2) sCOOH or -CH 2 S Y 3, Y 3 signifies a C 1 -C 4 alkyl group, benzyl may
substitute with methoxy or methoxy, or
CH3CONH-CH2-, o means an integer of 1 to 4,
A6 means Thr or D-Thr s means an integer of 3 to 5,
A8 represents the amino-acyl residue of a La-amino acid
Neutral lipophilic, Ag represents the amino-acyl residue of an L- or Da-amino
neutral lipophilic acid, and
Z1 means a polypeptide residue located in position 10
at 31 of a natural calcitonin or one of its
derivatives or analogues, which has a hypocalcemic activity, the 1 to 4 Y1 residues in the formula X may be identical or independently different, and, with the exception of the amino-acyl residue A8, all the amino acid residues in the formula X have the configuration
L or D as well as the salts and complexes of these compounds.

 Such compounds are described, for example, in British Patent Application 2,184,729, the contents of which are incorporated herein by reference.

 Z1 in formula X means a peptide residue which may be present at positions 10 to 31 in many known calcitonins, for example in human calcitonin, salmon, eel, chicken, beef, mutton, rat or pork, as well as in derivatives and analogues of these calcitonins having a similar activity. Derivatives and analogues of these calcitonins are especially natural calcitonins in which one or more amino acid residues have been replaced by one or more other amino acid residues, or in which the SS bridge has been replaced by an alkylene bridge, or in which one or more amino acid residues have been removed. These Z1 peptide residues normally contain 22 amino acids, but may also correspondingly contain a lesser number of amino acid residues by removal of one or more amino acid residues (des-aminoacyl derivatives).

Z1 preferably means a) Gly-Thr-Tyr-Thr-Gln-Asp-Phe-Asn-Lys-Phe-His-Thr
Phe-Pro-Gln-Thr-Als-Ile-Gly-Val-Gly-Ala b) Gly-Lys-Leu-Ser-Gln-Glu-Leu-His-Lys-Leu-Gln Thr-Tyr-Pro-Arg- Thr-Asp-Val-Gly-Ala-Gly-Thr c) Gly-Gly-Leu-SEr-Gln-Glu-Leu-His-Lys-Leu-Gln
Thr-Tyr-Arg-Pro-Thr-Asn-Thr-Gly-Gly-Ser-Thr
Compounds of formula X in which Z 1 has the definition of b) or c), preferably those in which Z 1 has the definition given under c), are especially preferred.

 R "CO is preferably the acyl residue of an aliphatic, cycloaliphatic, aromatic or heterocyclic carboxylic acid.

R "preferably means a) a linear or branched alkyl group, saturates or
unsaturated, containing from 1 to 17 carbon atoms,
especially an alkyl group saturates C3-Cg, b ') a C5-C7 cycloalkyl or cycloalkylalkyl group
in which the cycloalkyl residue is C5-C7
and the remainder C 1 -C 2 alkyl, c ') an adamantyl, adamantylmethyl or adaman group
tylethyl, or phenyl, benzyl or phenethyl.

In the definitions given above for
R ", the allyl, cycloalkyl or phenyl groups may contain usual substituents, for example halogens or N02, OH, alkoxy, e.tc.

 The radical R "CO may be, for example, the α-desamino residue or a natural α-amino acid.

For R ", the definitions a '), b') and c ') are preferred.

 Y 1 and Y 2 as residues on the α-amino acid of an α-amino acid represent in particular the residues which are bonded to the α-carbon atom of a natural α-amino acid, but can also means residues of other α-amino acids, for example 3-cyclohexylalanine or α-amino-isobutyric acid.

When o in the formula X is 4, a) the N-terminal aminoacyl residue (corresponding to
second residue of amirnacid in the sequence of calci
natural tones) preferably means Ser,
Gly or Ala, b) the second aminoacyl residue (corresponding to
third amino acid residue in the sequence
natural calcitonins) preferably means
Asn or Ser, c) the third amino-acyl residue (corresponding to
fourth amino acid residue in the sequence of
natural calcitonins) preferably means
Leu, Asn, Ser, Phe, D-Leu or the rest of the cyclo
hexylalanine, d) the fourth amino-acyl residue (corresponding to
fifth amino acid residue in the sequence
natural calcitonins) preferably means
Ser or Ala.

 When o in the formula X is 3, the N-terminal, second and third amino acid residues have the same preferred definitions as those given above for the case where o = 4 under b).

 When o in formula X means 2, the N-terminal and the second amino acid residue have the same preferred definitions as those given above for the case where o = 4 under c) and d).

 When o in the formula X means 1, the N-terminal and the second amino acid residue preferably signify Ser or Ala.

signifie de préférence Cys, un dérivé de la cysteine tel que donne ci-dessus pour Y2 ou un reste a-amino-acyle lipophile neutre, spécialement Ala ou un autre reste a- amino-acyle lipophile neutre, tout particulièrement Ala.A6 means of preference Thr

is preferably Cys, a cysteine derivative as given above for Y2 or a neutral lipophilic α-amino-acyl residue, especially Ala or other neutral lipophilic α-amino-acyl residue, most preferably Ala.

A8 is preferably the aminoacyl residue of a neutral lipophilic α-amino acid, especially
Val or Gly.

 Ag also preferably means the amino-acyl residue of a neutral lipophilic α-amino acid, especially Leu or Phe.

 In the compounds of formula X, o is preferably 2, where R is H or R "CO, or in particular o is 1 and R is R" CO.

 All amino acid residues preferably have the L configuration.

Glucosylated calcitonins (including derivatives and analogues) are especially those which are glucosylated on the N-terminal amino group or on one or more amino groups located in one or more side chains, for example the compounds of formulas XIa to XIf

or
HOH2C- (CHOH) -Cr-CH2NH-Calc (XIf) in which Calc represents the remainder of a natural calcitonin or a derivative or analog thereof, linked to the carbohydrate residue through an amino group on N-terminus or in a side chain.

 The calcitonin derivatives of formula X can be prepared according to the generally known methods for the synthesis of polypeptides of this type.

ou CH2-S-S-CH2-CH2-COOH. on fait réagir un compose de formule XII

sous forme protégée ou non protégée, avec un composé de formule XIII

dans laquelle R10 représente un groupe qui facilite la formation d'un pont S-S- avec l'atome de soufre et l'autre groupe CH2SH présent dans le peptide de formule XII, R11 signifie l'hydrogène ou un groupe protecteur du groupe amino,
R12 signifie OH ou un groupe protecteur du groupe carboxy , et
V signifie l'hydrogène ou un groupe NH, ou bien on fait réagir un composé de formule XIY

sous forme protégée ou non protégée, dans laquelle
R10 a la signification donnee précédemment, avec un composé de formule XV

The polypeptides corresponding to the above formulas can be prepared, for example, as follows: a) at least one protecting group present is eliminated;
in a protected polypeptide having the sequence
given in the formula X, or b) two pairs of peptides
each of which contains at least one amino acid residue or
one of its derivatives, as described for the formula X, s6us
protected or non-protected form, the peptides
diques being such that we obtain the sequence
of amino acids corresponding to formula X and,
if necessary, step a) of
process, or c) reacting a compound of formula X, wherein
R means hydrogen, in protected form or
unprotected, with an acidic formula R "COOH or one
reactive derivatives and, if necessary,
step a) of the process, or d) to prepare a compound of formula X wherein Y2, signifies

or CH2-SS-CH2-CH2-COOH. reacting a compound of formula XII

in protected or unprotected form, with a compound of formula XIII

wherein R10 represents a group which facilitates the formation of an SS- bridge with the sulfur atom and the other CH2SH group present in the peptide of formula XII, R11 signifies hydrogen or a group protecting the amino group,
R12 is OH or a protecting group of the carboxy group, and
V is hydrogen or NH, or a compound of formula XIY is reacted

in protected or unprotected form, in which
R10 has the meaning given above, with a compound of formula XV

and optionally the step a) is implemented
of the process.

 When the preparation of the starting materials is not described, these compounds are known or can be prepared and purified according to known methods.

The end products of formula X can be purified by the usual methods so that they contain less than 5% of polypeptide by-products.

The peptides used as starting materials in steps a) and b) of the process can similarly be prepared in a known manner in solution or in the solid phase method.

 Peptide units which contain a -CH 2 -S -S-CH 2 -CH 2 -COOH or CH 2 -SS-CH 2 -CH (NH) 2 -COOH group as the Y 2 - residue may be prepared analogously to step d) the previously mentioned method.

ou S-SO3
Dans ces restes, alkyle signifie spécialement un groupe alkyle inferieur contenant de 1 à 4 atomes de carbone. L'introduction de ces restes dans les composes ayant des groupes SH libres peut etre effectuée de maniere analogue aux méthodes connues dans la chimie du soufre.In this step of process d), compounds of formula XIII or XIV are used in which
R10 means known residues which react with thiols while forming an SS- bond. R10 especially S-alkyl, -S-CoOalkyl,

or S-SO3
In these residues, alkyl signifies especially a lower alkyl group containing from 1 to 4 carbon atoms. The introduction of these residues into the compounds having free SH groups can be carried out analogously to the known methods in sulfur chemistry.

 Another preferred class of compounds of the invention comprises a group of LH RH antagonists.

Preferably, the compounds comprise the glucosyl derivatives of the compounds of formula XVI
R1-A1-E1-C1-D1-E1-F1-G1-H1-I1-K1-NE2 (XVI) wherein R1 is H or a C1-C7 acyl group;
A1 is D-Phe, optionally substituted in the phenyl ring by F, C1, Br, NH2, CH3 or OCH3, especially in the 4-position, a 5-D-naphthylalanine residue,
D-Trp optionally substituted in position 5 or 6 by
F, C1, Br, NH2, or OCH3 and / or substituted in the 1-position by a formyl or acyl group, a proline residue, 3,4-dehydroproline or D-pyroglutamine
B1 means D-Phe optionally substituted with F, C1,
Br, NH2, CH3 or CH30 in the phenyl ring,
D- α -methylphenylalanine, optionally substituted
in position 4 by C1, or
ss-D-naphthylalanine.

C1 means D-Trp possibly substituted in position
5 or 6 by F, C1, Br, NH2 and / or OCH3 and / or in position
1 by HCO or CH3CO,
ss-D-naphthylalanine,
3-D-pyridylalanine,
D-Tyr, or
Phe optionally substituted with F, C1, Br, NH2, CH3, or
CH 3 O,
D1 means Ser,
E1 means Tyr or optionally substituted phenylalanine
in the phenyl ring by Cl, Br, NH2, CH3 or CH30,
F1 means D-Phe, optionally substituted in the
phenyl ring with F, C1, Br, NH2, CH3 or CH30,
D-Trp possibly substitute in position 5 or 6
by F, Cl, Br, NH2 or CH30 and / or in position 1 by
formyl or acetyl,
D Tyr,
SD-naphthylalanine, D-Leu, D-Ile, D-Nle, DVal, D-Ser
(OtBu), D-Arg, optionally dialkylated by
C 1 -C 6 alkyl or C 5 -C 6 cycloalkyl groups,
D-homoarginine optionally dialkylated by
C1-C6 alkyl or C5-C6 cycloalkyl groups,
D-His, D-His (Bzl),
D-Lys or D-Orn, both possibly dialkylated
with C 1 -C 6 alkyl or C 1 -C 6 cycloalkyl groups,
D-Phe (p-NH2) or
ap-aminocyclohexylalanine,
G1 is Leu, Nle, Nva, Na-methylleucine, Trp,
Phe, Met or Tyr,
H1 means Arg, Lys or Orn possibly substituted
with C 1 -C 6 alkyl or C 5 -C 6 cycloalkyl groups,
I1 means Pro, hydroxyproline, or 3,4-dehydroproline,
and
K1 means D-Ala.

 If desired, E1 and F1 can be replaced by each other. D1 and K1 may, if desired, mean Cs-residues linked by an S-S- bridge.

 If desired, one of the symbols D1 and K1 means Asp or Glu and the other means Orn, diaminopropionic acid or diaminobutyric acid, and the radicals D1 and K1 are linked by an amide bridge.

The preferred meanings are the following
R1 = acetyl or formyl
A1 = O-Phe, D-Phe (p-Cl), s-D-naphthylalanine, 3,4-dehydropyridine,
B1 @ D-Phe eventually substituted as indicated
previously,
C1 = D-Trp possibly substituted as indicated
previously,
D1 = Ser
When the rest is substituted, it is preferably mono-substituted.

(i) E1 = Tyr or Phe possibly substitute as
indicated previously, when F1 = D-Phe or Lys,
or (ii) E1 = D-Phe or Lys when F1 = Tyr or Phe if any
substitutes as indicated above,
G1 = Leu
H1 = Arg I1 = Pro
K1 = D-Ala
In the mentioned LHRH antagonists
above, the carbohydrate residue is preferably
attached to the N-terminal amino group or to a free amino group located in a side chain of the peptide.

Glucosyl derivatives preferably have the following structures XVIa to XVIf in which NH2-LHRH
antag. "means an LHRH antagonist
of formula XVI given previously:

G3-CO-NE-LHRE-Antag. (XVIc)

In formulas XVIa to XVIf above,
for the sake of simplicity, it has been shown
only a carbohydrate residue linked to an amino group.

However, the molecule may contain more than one residue
carbohydrate when the LHRH antagonist contains
several amino groups. Preferably, the molecule
contains two carbohydrate residues.

LHRH antagonists, used as
starting materials, are known or can be prepared
according to known methods, for example as described
in European Patent Applications No. 81887 and
201260.

Other preferred glucosylated derivatives are
those derived from the following peptides
(a) oxytocin and vasopressin, and their
derivatives, for example Lys8-vasopressin and
Orn8 vasopressin,
(b) insulin,
(c) GRF and its derivatives.

The starting materials and the compounds of
the invention can be prepared synthetically in
liquid phase or solid phase.

The compounds of the invention can be
advantageously prepared by solid phase synthesis.

The Applicant has found a special process
advantageously for the preparation of peptide-alcohols which comprise at the C-terminus of the channe
peptide two groups alcohol or alcohol group
and a thiol group. The process is specially
suitable for the preparation of peptide-alcohols
containing a C-terminal threoninol, serinol or
cysteinol.

 Examples of suitable compounds include some of the somatostatin described above.

Solid phase peptide synthesis has been shown to be a particularly fast and favorable process for the preparation of peptides and has therefore become a generally method.
common.

 As is known, an amino acid is first attached by its carboxy groups to the hydroxy or amino group of an insoluble synthetic resin to form an ester or amide group; the other amino acids are then added to this resin in the desired order and the final polypeptide is cleaved from the carrier resin.

This synthesis is carried out without any problem for normal polypeptides having amino acids
C-terminal. However, polypeptide-alcohols which have at the C-terminus an amino alcohol instead of an amino acid, do not readily form a bond with carrier resins having OH or NH 2 groups and / or are not easily split when the synthesis is complete.

The following processes have been previously proposed as solid phase processes for the preparation of peptide alcohols: a) conventional preparation of the corresponding polypeptide
containing at the C-terminus an amino acid
(as an ester with a resin having
OH groups) and then sci'ssion by reduction in
using boron hydrides, the carboxy group
being converted simultaneously into alcohol
(U.S. Patent No. 4,254,023 / 4).

(b) Addition of the terminal amino alcohol in the form
of ether with a hydroxymethylated resin using
carbonyl diimidazole and, after synthesis of the
peptide, split using HC1 / TFA or HBr / TFA
(Kun-hwa Hsieh and GR Marshall, ACS National
Meeting, New Orleans, 21-25, 3. 1977).

 However, these two methods require energetic splitting conditions.

We have found that the cleavage of the peptide from the resin, while simultaneously forming the C-terminal peptide alcohol, is carried out under mild conditions when the C-terminal amino alcohol is bound to the resin by an acetal bond.
According to the invention, the peptide-alcohol which comprises at the C-terminal end of the peptide chain two alcohol groups or an alcohol group and a thiol group, is prepared by acid hydrolysis of an acetal of the peptide-alcohol with a resin. polymer containing formylphenyl groups (designated synthesis of the invention in the present application).

The reaction can be schematically illustrated as follows

or
is the rest of an insoluble synthetic resin
Z represents a direct bond or a remainder that connects
the resin with the acetylphenyl group acetalized
X means O or S
R1 is hydrogen or methyl, and
Y is the rest of a peptide-alcohol that can for example
have protective groups,
the group CHO- possibly acetalized being
located in meta or para position of the rest Z.

For reasons of simplicity, in
formulas Iret ID of the scheme above, we have
only one substituent on the resin; it is obvious
however a number of these groups are
related to the molecule of the polymer resin. The split
peptide-alcohol from the resin, by hydrolysis
of the acetal group, is carried out as mentioned above
under acidic conditions, for example with
diluted trifluoroacetic acid. Hydrolysis can
be carried out at room temperature.

When Z in the formula 1r signifies a
direct bond, the phenyl radicals containing the
Acetal groups are directly related to the polymer residue
and belong to the polymer. Examples of such compounds of formula I, r are the acetals of a
formylated polystyrene resin (in the formula Ir, # in this case means a polyethylene chain).

When Z means a remainder, this remains contains
in this case a group that results from the reaction
a reactive group, linked directly or indirectly
to the polymer, with another reactive group directly linked
or indirectly to the formylphenyl (acetalized) group.

The remainder Z can be represented for example by the following formula IIIr:
- (D) P-Q1-Q2- (E) q- (IIIr) in which
Q1 = the rest of a reactive group linked to the polymer
Q2 = the rest of a reactive group linked to the group
formylphenyl (acetalized)
D = a remainder that links the Q1 group to the polymer
E = a remainder that connects the group Q2 with the
formylphenyl group (acetalized) p and q, independently of each other, means O or 1.

 The group Q1-Q2 preferably means an ester or amide group, especially a carboxamide group. Q1 is preferably NH and Q2 is preferably CO.

signifie un reste de formule IVr

dans laquelle
R = l'hydrogène ou un groupe méthyle et m signifie O ou 1, le groupe acétal etant situé en position méta ou para.D and E, independently of one another, include, for example, alkylene or alkyleneoxy radicals containing from 1 to 5 carbon atoms. Compounds of formula Ir in which Z denotes a radical of formula IIIr are, for example, compounds in which D-Q1 is the residue of an aminomethylated polystyrene resin and the remainder

means a remainder of formula IVr

in which
R = hydrogen or a methyl group and m means O or 1, the acetal group being located in the meta or para position.

et t signifie le polystyrène.In this case, Z represents therefore

and t is polystyrene.

The remainder IVr preferably means a remainder

 Instead of polystyrene aminomethyl, other polymers can also be used, especially those containing NH 2 groups, for example polyacrylamides containing aminoethyl groups.

 As mentioned above, the acetyl formylphenyl group is preferably bonded to the polymer by an amide bond. This ensures that binding of the formylphenyl residue acetalized to the resin is stable during synthesis and scission of the polypeptide and that cleavage takes place on the acetal bond as desired so that on the one hand the peptide alcohol is synthesized and that on the other hand, the formylphenyl residue remains on the resin.

 If desired, the peptide alcohol may be attached at a greater distance from the resin by incorporation of a spacer moiety (spacer) between the reactive groups of the polymer (especially amino groups) and the reactive groups. formalised acetaldehyde derivative (especially carboxy groups). For certain reactions on the alcohol peptide, this can advantageously take place before the cleavage (for example oxidation of the cysteine residues). In this case, the residue D or E in the formula IIr additionally contains the spacer group and Q1 or Q2 represent the reactive residue of the spacer group.

 The spacer group may be, for example, an aminocarboxylic acid, such as 5-aminocaprolic acid.

In a specific case, when using aminomethyl polystyrene, a remainder of formula IVr and amino-amino acid as spacing group, Z stands for

dans laquelle A représente un groupe protecteur du groupe amino et le groupe acétal est en position meta ou para du reste Z.A cet effet, on scinde d'abord le groupe protecteur A puis on fait réagir le groupe amino libre avec l'amino-acide N-protégé suivant etc.., jusqu'à ce que tous les amino-acides aient éte ajoutés a la resine dans la sequence correspondant à celle du peptide-alcool désiré.The compounds of formula Ir can be prepared according to methods which are usual in solid phase technology, using as starting material the compound of formula Vr

wherein A represents a protecting group of the amino group and the acetal group is in the meta or para position of the remainder ZA for this purpose, the protecting group A is first cleaved and then the free amino group is reacted with the amino acid N -protected next etc., until all amino acids have been added to the resin in the sequence corresponding to that of the desired peptide-alcohol.

The protecting groups of the amino group which must be chosen for the amino acids used
or for the aminoalcohol, should be those which are cleaved under non-acidic conditions, since under acidic conditions a hydrolysis of the acetal group takes place. The group
CF3CO- or the group FMOC- (9-fluorenylmethyloxycarbonyl) may be used, for example as protecting groups of the amino group. These protecting groups are cleaved in a basic medium according to the methods known in peptide chemistry.

 Only the protecting groups located in the side chain and the protecting group of the amino group of the last introduced amino acid can be acid labile and are therefore removed simultaneously from the resin with the release of the alcohol peptid.

 Preferably, the Boc groups are present as protecting groups.

 As bases, KOH or piperidine or NaBH 4 is preferably used.

The synthesis of the peptide ring can be carried out in a conventional manner from a
peptide residue having free amino groups and an amino acid whose carboxy group is free or activated.

 The reaction can be carried out by adding, for example, hydroxybenzotriazole and dicyclohexylcarbodiimide.

The compounds of formula Vr can be prepared for example a) by reaction of a resin comprising a group
aldehyde and corresponding to the formula IIr

avec un composé de formule VIr

dans laquelle groupe acétal est en position meta ou para du groupe Q1, -(E)- q et Q; et Q2, sont deux groupes réactifs qui reagissent ensemble pour former un pont Q1 - Q2. in which the CHO group is in the meta position
or para of the substituent Z,
with a N-protected amino alcohol of formula HX-CHR1-CH (NHA) -CH20H
optionally in activated form, or b) by reaction of a resin of formula

with a compound of formula VIr

wherein the acetal group is in the meta or para position of the group Q1, - (E) - q and Q; and Q2, are two reactive groups that react together to form a Q1 - Q2 bridge.

 The acetalization of process a) can be carried out in the presence of an acid as a catalyst.

Suitable acids include p-toluenesulfonic acid and p-trifluoromethylphenylsulfonic acid.

 If desired, the trimethylsilyl group can be used as a protecting group for a free alcohol.

 The esterification process b) can be carried out under very mild conditions, for example by reacting a derivative of a carboxylic acid with a polymer having OH or NH 2 groups.

avec un composé de formule HX-CHR1 -CH(NHA)-CH2OH
-t'acetalisation peut être effectuée comme indiqué dans le procede a).The compounds of formula VIr can be prepared by acetalizing a compound of formula

with a compound of formula HX-CHR1 -CH (NHA) -CH2OH
Acetalisation can be carried out as indicated in the method a).

 During the synthesis and cleavage of the alcoholic peptide from the resin, other reactions may be carried out, for example removal of the protecting groups, for example removal of thiol protecting groups, or oxidation of the cystine residues.

 Such reactions can be carried out after cleavage of the peptide-alcohol in the liquid phase.

According to the synthesis of the invention, pharmacologically active peptides and other peptides which contain on the C-terminal end 2 alcohol groups or an alcohol group and a thiol group can be prepared with ease.
The following examples illustrate the present invention without in any way limiting its scope.

All temperatures are given in degrees Celsius and the values [a] D are uncorrected. The following abbreviations have been used:
AcOH = acetic acid
Boc = tert-butoxycarbonyl
But = tert.-butyl
DCCI = dicylcohexylcarbodiimide
DMF = dimethylformamide
Fmoc = 9-fluorenylmethoxycarbonyl
MeOH = methanol
NEt3 = triethylamine
Thr-ol = threoninol residue = CH3-CHOH-CH (CH20H) -NH
TFA = trifluoroacetic acid
HOBt = N-hydroxybenzotriazole hpGRF = factor that triggers the secretion of the hormone
human pancreatic growth
HOSu = N-hydroxy-succinimide.

 All peptides are obtained as polyacetate polyhydrate having a peptide content of 70 to 90%.

 Liquid chromatography (HPLC) indicates that the peptides contain less than 5% of other peptides.

 The factor "F" mentioned in the following examples indicates the peptide content in the product obtained (F = 1 indicates a peptide content of 100%).

The difference up to 100% (<1-F) x 100] consists of acetic acid and water.

 All sugars have the configuration unless indicated otherwise.

acétate

Example 1 N &alpha; -ss-deoxyfructosyl

acetate

on ajoute 3 ml d'acide trifluoroacétique (100%) et on maintient à la température ambiante jusqu'à ce que tout le produit de départ soit dissous (5 minutes). Après addition de 20 ml d'éther diisopropylique, on sépare par filtration le composé du titre qui a precipité et on le lave à l'éther diisopropylique. Le composé est purifié par chromatographie sur gel de silice (eluant : CHC13/MeOH/HOAc/H20 7/3/0.5/0.5) et isole sous forme d'un lyophilisat blanc. [&alpha;]D20 = -31,3 (c= 0,52 dans HOAc à 95%)
F : 0,88.Acetate
400 mg of [N-alpha-deoxyfructosyl] -D-Phe-

3 ml of trifluoroacetic acid (100%) are added and the mixture is kept at room temperature until all of the starting material has dissolved (5 minutes). After addition of 20 ml of diisopropyl ether, the precipitated title compound was filtered off and washed with diisopropyl ether. The compound is purified by chromatography on silica gel (eluent: CHCl 3 / MeOH / HOAc / H 2 O 7/3 / 0.5 / 0.5) and isolated in the form of a white lyophilizate. [&alpha;] D20 = -31.3 (c = 0.52 in 95% HOAc)
F: 0.88.

The starting product can be prepared as follows:

To a solution of 10 g of

acétate dans 10 ml de DMF, on ajoute lentement goutte à goutte à la température ambiante 2,25 g de percarbonate de di-tert.-butyle dissous dans 30 ml de DMF. Après deux heures à la temperature ambiante, on élimine le solvant sous vide et on ajoute 200 ml d'éther diisopropylique au résidu. Le produit qui s'est formé est séparé par filtration, lave à l'éther diisopropylique et séché.Le produit brut est purifie par chromatographie sur gel de silice (éluant : CH2C12/MeOH 9/1) et ensuite isole sous forme d'une poudre amorphe blanche. [&alpha;]D20=29,8 (C = 1,28 dans le DMF), b) N&alpha;-ss-désoxyfructosyl

In 2.25 g of di-tert.-butyl percarbonate dissolved in 30 ml of DMF, 2.25 g of di-tert.-butyl percarbonate are slowly added dropwise at room temperature dropwise at room temperature. After two hours at room temperature, the solvent was removed in vacuo and 200 ml of diisopropyl ether was added to the residue. The product which has formed is separated by filtration, washed with diisopropyl ether and dried. The crude product is purified by chromatography on silica gel (eluent: CH 2 Cl 2 / MeOH 9/1) and then isolated in the form of a white amorphous powder. [&alpha;] D20 = 29.8 (C = 1.28 in DMF), b) N &alpha; -ss-deoxyfructosyl

 2 g of D - (+) - glucose and 0.5 g of the compound of step a) above are dissolved in 20 ml of a 9/1 (vol / vol) mixture of MeOH and HOAc. and kept at 60-70 for three hours. After evaporation, the product is taken up in a little methanol and the compound is precipitated by addition of diisopropyl ether. The title compound thus obtained is purified by chromatography on silica gel (eluent: CH 2 Cl 2 / MeOH 9/1).

The product is obtained in the form of an amorphous powder.

 [&alpha;] D20 = 12.0 (c = 1.04 in DMF).

By proceeding in a manner analogous to that described in Example 1, the acetates of the following compounds are obtained in which SMS signifies the polypeptide residue of formula

Example 2
N &alpha; - [alpha &; -glucosyl (1-4) -désoxyfructosyl) -SMS

As a starting product, use the
D (+) - maltose instead of D-glucose. [&alpha;] D20 -7.90
(c = 0.71 in 95% AcOH) F: 0.91.

Example 3 N &alpha; - [&alpha; -glucosyl (1-4) - &alpha; -glucosyl (1-4)
fructosyl SMS

F: 0.78
As a starting material, maltotriose is used instead of D-glucose [& alpha] D20 = +11.3 (c = 0.71 in 95% AcOH).

Example 4 N &alpha; fructofuranuronic acid-SMS

F: 0.85
As a starting material, the acid is used
D-glucuronic in place of D-glucose. [&alpha;] D20 = -29.4 (c = 0.34 in 95% AcOH).

Example 5 N-deoxysorbosyl-SMS

F: 0.85
As a starting material, we use the
D (+) - galactose instead of D-glucose. [&alpha;] D20 = -30.4 (c = 0.50 in 95% AcOH).

Example 6 - [O-ss-D-glucosyl- (1-4) -deoxyfructosyl] -SMS

F: 0.81
As a starting product, we use the
20
D (+) - cellobiose. [&alpha;] D20 = -28.1 (c = 0.47 in 95% AcOH).

Example 7 N &alpha; -L (-) - deoxyfructosyl-SMS

F: 0.91
As a starting product, we use the
L (-) - glucose instead of D (+) - glucose.

 [&alpha;] D20 = -20 (c = 0.46 in 95% AcOH).

Example 8 N- [O-ss-D-glucosyl- (1-6) -desoxyfructosyl] -SMS

 As starting material, gentiobiose is used instead of D-glucose.

F : 083
Comme produit de départ, on utilise le
D(+)-Tactose à la place du D-glucose. [&alpha;]D20 = -29,3 (c = 0,55 dans AcOH à 95%).[&alpha;] D20 = 23.5 (c = 0.46 in 95% AcOH) F: 0.76
Example 9 N- [O-ss-D-galactosyl- (1-4) -desoxyfructosyl] -SMS

F: 083
As a starting material, we use the
D (+) - Tactose instead of D-glucose. [&alpha;] D20 = -29.3 (c = 0.55 in 95% AcOH).

Example 10 - (O- &alpha; -galactosyl- (1-6) -desoxyfructosyl) -SMS

 As a starting material, melibiose is used instead of D-glucose.

 [&alpha;] D20 = +8.4 (c = 0.5 in 95% AcOH) F: 0.76.

Example 11 [N- (1-Deoxy-D-fructosyl) -Try] -SMS

As a starting material, we use the
20
Tyr3-SMS instead of SMS [&alpha;] D20 = - 32.2 (c = 0.9 in 95% AcOH) F: 0.87.

Example 12 [N - (α-D-glucopyranosyl- (1-4) -1-deoxy fructosyl), Tyr] -SMS

F = 0.81
As a starting material, we use the
D (+) - maltose instead of D-glucose and Tyr3-SMS instead of SMS. [&alpha;] D20 = 4.7 (c = 1.0 in 95% AcOH)
Example 13

 As a starting material, Dglucoheptose is used instead of D-glucose.

 [&alpha;] D20 = - 12.9 (c = 1.0 in 95 AcOH) F = 0.91.

Example 14

As a starting product, we use the
D (+) - glucose and unprotected SMS on the # -NH2 group of lysine.

[&alpha;] D20 = -42.4 (c = 0.37 in 95% AcOH). F = 0.83
Example 15

 As a starting material, glucoheptose and unprotected SMS are used on the # -NH 2 group of lysine.

 [&alpha;] D20 = -9.3 (c = 0.41 in 95% AcOH). F = 0.84.

Example 16
Fructosyl - 6 - phosphate - SMS

As a starting material, we use the
D-glucose-6-phosphate in place of D-glucose.

[&alpha;] D20 = 19.5 (c = 1.0 in 95% AcOH). F = 0.89,
Example 17

As a starting product, we use the
D-ribose instead of D-glucose. [&alpha;] D20 = -31.8 (c = 1.0 in 95% AcOH).

Example 18 N α -désoxyfructosyl- (D) Phe-Cys [COC (CH 3) 3] -
Phe- (D) -Trp-Lys-Thr-Cys [COC (CH3) 3] -Thr-ol a) Deoxyfructosyl- (D)

Thr-ol -----------------------
0.58 g of the compound of Example 1 in 10 ml of DMF are mixed with 0.08 ml of NEt3 and then with 0.12 ml of (BOC) 20. The mixture is stirred for about 15 hours at room temperature, concentrated in vacuo and stirred into ether. The precipitated product is filtered off. The residue is dissolved in a little methanol and the product is precipitated by the addition of water. We filter the product, we wash it
with a little water and we dry it, which gives the
composed of the title.

 [&alpha;] D20 = +14.5 (c = 0.7 in DMF).

b) N-alpha-oxyfructosyl- (D) -Phe'-Cys-Phe- (D) Trp-Lys (BOC) -Thr
Cys-Thr-ol
0.51 g of the compound of step a) above
in a mixture of 10 ml of dioxane and 2 ml
of buffer NEt3 / AcOH at pH 8.6 is added under argon
a total of 0.4 g of dithioerythritol. We shake the
mixture for about 15 hours at room temperature
room temperature and concentrated in vacuo. We separate by
centrifugation the product that precipitated, washed
the residue with a little water and dried under vacuum.

 The title compound is thus obtained.

 [&alpha;] D20 = +3.8 (c = 0.8 in DMF).

c) N &alpha; -désoxyfructosyl- (D) Phe'-Cys (COC (CH3) 3] -Phe- (D) Trp-Lys-
(BOC) -Thr-Cys (COC (CH3) 3] -Thr-ol
0.38 g of the compound of
step b) above in 25 ml of N-methylpyrrolidone,
0.3 ml of N-methylmorpholine and 0.31 ml are added to 0.
of pivaloyl chloride, shake for about
16 hours to 0. and we stir in a melanae
of ether and diisopropyl ether. We separate by
centrifugation the product that precipitated. The residue
is dissolved in a little DMF and is made to precipitate
the product by addition of methanol and water. The
product is centrifuged, dried under vacuum
and uses for the next reaction without any other
purification.

d) N &alpha; -désoxyfructosyl- (D) Phe-Cys [COC (CH3) #] - Phe- (D) Trp-Lys-
Thr-Cys [COC (CH3) 3] -Thr-ol
The residue of step c) is dissolved at 0
above in 5 ml of a 9: 1 mixture of TFA and
of water and stirred for 15 minutes. We do
precipitate the product by adding a mixture
ether / 10% HCl 5N / etner. We filter the product, we
wash with ether and dry. The residue is purified
by chromatography on silica gel with a
CHCl3 / MeOH / AcOH / H2O mixture. We bring together
fractions containing the desired product, they are
vacuum center while adding water then one
lyophylise. This gives the compound of
title. [&alpha;] D20 = -15.3 (c = 1.0 in 95% AcOH)
F: 0.88.

désoxy-D-glucosa

Example 19

deoxy-D-glucose

(préparé comme decrit à l'exemple la) dans 100 ml d'un mélange 9/1 de MeOH et de AcOH et on maintient a 65 pendant 16 heures. Après évaporation, on dissout le produit dans un peu de méthanol et on fait précipiter le produit avec de l'éther diisopropylique. Pour eliminer le groupe protecteur (scission du groupe BOC),on utilise le produit brut ainsi obtenu, sans purification prealable. 2 g of D - (-) - fructose and 1 g of

(prepared as described in Example 1a) in 100 ml of a 9/1 mixture of MeOH and AcOH and maintained at 65 for 16 hours. After evaporation, the product is dissolved in a little methanol and the product is precipitated with diisopropyl ether. To eliminate the protecting group (cleavage of the BOC group), the crude product thus obtained is used without prior purification.

 1 g of the crude product thus obtained is mixed with 20 ml of trifluoroacetic acid (100%) and maintained at room temperature until the starting material is dissolved (5 minutes). 200 ml of diisopropyl ether are then added, the precipitated product is filtered off and washed with diisopropyl ether. The title compound is purified by chromatography on silica gel (eluent CHCl 3 / MeOH / AcOH / H 2 O 7/3 / 0.5 / 0.5) and is isolated in the form of a white lyophilizate.

 Lava = -6.7 (c = 0.3 in 95% AcOH) F: 0.73.

A second product, the following 1: 1 mixture of isomers having the reverse configuration on the C2 carbon atom of the carbohydrate residue, can be obtained:

Example 20

 By proceeding in a manner analogous to that described in Example 19 and using Tyr-SMS as the starting product instead of SMS, the title compound is obtained.

 [&alpha;] D20 = -2.9 (c = 1.0 in 955 AcOH) F: 0.95.

Example 21
Glucuronamide from

par 3 ml d'acide trifluoroacétique à 100% jusqu'à obtenir une dissolution complète (5 minutes). On fait précipiter le composé du titre sous forme de trifluoroacétate par addition de 20 ml d'éther diisopropylique. Après filtration, séchage et chromatographie sur gel de silice (éluant : CHC13/MeOH/AcOH/H20 7/3/0,5/0,5), on obtient le composé du titre a l'état pur sous forme d'un lyophilisat (acetate).170 mg of the glucuronamide of the

with 3 ml of 100% trifluoroacetic acid until complete dissolution (5 minutes). The title compound is precipitated as trifluoroacetate by addition of 20 ml of diisopropyl ether. After filtration, drying and chromatography on silica gel (eluent: CHCl 3 / MeOH / AcOH / H 2 O 7/3 / 0.5 / 0.5), the pure title compound is obtained in the form of a lyophilizate. (acetate).

 [&alpha;] D20 = -29.2 (c = 0.48 in 95% AcOH) F: 0.85.

117 mg d'acide glucuronique et de 135 mg de HOBT dans du diméthylformamide, on ajoute une solution à 135 mg de DCCI dans 2 ml de DMF . Après 48 heures avec décongelation simultanée à la température ambiante, on sépare la dicyclohexylurée par filtration et on fait précipiter le composé du titre par addition de 20 ml d'éther diisopropylique.Après filtration, sechage et chromatographie sur gel de silice (eluant : CH2C12/
MeOH 9:1), on obtient le composé du titre a l'état 20 @@@@@@@@@@@@@@@@@@@@@@
Exemple 22
Quinamide du

The starting product can be obtained as follows
To a solution cooled to -30 C of 450 mg of

117 mg of glucuronic acid and 135 mg of HOBT in dimethylformamide, a solution of 135 mg of DCCI in 2 ml of DMF is added. After 48 hours with simultaneous defrosting at room temperature, the dicyclohexylurea is filtered off and the title compound is precipitated by the addition of 20 ml of diisopropyl ether. After filtration, drying and chromatography on silica gel (eluent: CH 2 Cl 2 /
MeOH 9: 1), the title compound is obtained in the following manner:
Example 22
Quinamide from

 By proceeding in a manner analogous to that described in Example 21 and using L (-) - quinic acid as starting material, the title compound is obtained.

 [&alpha;] D20 = -50 (c = 0.44 in 95% AcOH) F: 0.97.

Example 23
Sialamide of

 By proceeding in a manner analogous to that described in Example 21 and using as starting material sialic acid, the title compound is obtained.

 [&alpha;] D20 = -60.8 (c = 0.6 in 95% AcOH) F: 0.95.

Example 24 N &α; - (O-ss-D-glucoxyloxyacetyl) -DPhe-

250 mg of (tetra-D-acetyl-O-ss-
D-glucosyl) -oxyacétyl

Thr-ol in 10 ml of methanol and is adjusted to a pH of
10 with a few drops of a 1N solution of NaOCH3
in methanol. After reaction for 15 minutes,
the solution is neutralized with a resin exchange
ion (eg AMBERLYST # 15, H +) and separates
the resin by filtration. The filtrate is concentrated
and the residue is treated for 5 minutes with 3 ml
of trifluoroacetic acid. We make the
compound of the title as trifluoroacetate
by addition of 20 ml of diisopropyl ether and is isolated in pure form as a white lyophilisate
after filtration, drying and chromatography on
silica gel (eluent: CHCl3 / MeOH / AcOH / H20 7/3 / 0.5 / 0.5).

[α] 20 = -39.20 (c = 0.60 in 95% AcOH) F: O, 91.

D
The starting product can be prepared as
a) Benzyl ester of tetra-O-acetyl-O-s-D-acid
glucosyl-glycolic
To a solution of 830 mg of
benzyl in 50 ml of CH 2 Cl 2, 2.5 g of
4 A molecular sieve pulverized and, after addition
2.8 g of silver trifluoromethanesulfonate,
a solution of 4.1 g is added dropwise
of acetobromoglucose in 50 ml of CH2Cl2 after
15 minutes, the reaction is stopped by addition
4 ml of pyridine, the solid products are separated
by filtration and the filtrate is extracted with a
10% solution of NaHSO4. The title compound
is isolated in its pure state after chromatography on
silica gel (eluent: 99/1 CH2Cl2 / MeOH).

 [&alpha;] D20 = -22.4 (c = 1.7 in CHCl3).

(b) tetra-O-acethyl-O-ss-D-glucosyl-glycolic acid
800 mg of benzyl ester are dissolved
tetra-O-acethyl-O-ss-D-glucosyl-glycolic acid
in 40 ml of a mixture of equal parts by volume
of ethanol and water, 400 mg of
10% palladium and hydrogen under
3.45 bar in a "PARR" apparatus.
titre is isolated in the crystallized state after filtra
concentration and concentration under vacuum.

 [&alpha;] D20 = -35.5 (c = 1.03 in MeOH).

c) N &alpha;-( tetra-O-acetyl-O-ss-D-glucosyloxyacetyl) -DPhe-

To a solution cooled to -30 C and consisting of 81 mg of tetra-O-acetyl-O-s-D-gucosylglycolic acid, 225 mg of

et de 45 mg de HOBT dans 2 ml de DMF, on ajoute 45 mg de DCCI dissous dans 1 ml de DMF.

and 45 mg of HOBT in 2 ml of DMF, 45 mg of DCCI dissolved in 1 ml of DMF are added.

After 48 hours and after warming to room temperature, the dicyclohexylide is filtered off and 200 ml of diisopropyl ether are added to the filtrate to precipitate the title compound.

By proceeding in a manner analogous to that described in Example 24, the following compounds can also be prepared in which SMS signifies the polypeptide residue of formula

[&alpha;]D20 = -37.5 (c = 1 dans AcOH à 95%) F : 0,95
Exemple 26 N&alpha;-(O-ss-Callobiosyl-oxyacéthyl)-SMS

[&alpha;]D20 = -32,5 (c = 1 dans AcOH à 95%) F = 0,91.Example 25
N &alpha;-( O-ss-D-glactoxyl-oxyacéthyl) -SMS

[&alpha;] D20 = -37.5 (c = 1 in 95% AcOH) F: 0.95
Example 26 N &alpha;-( O-ss-Callobiosyl-oxyacetyl) -SMS

[&alpha;] D20 = -32.5 (c = 1 in 95% AcOH) F = 0.91.

[&alpha;]D20 = -32,9 (c = 1, dans AcOH à 95%) F : 0,93
Exemple 28
N&alpha;-(O-&alpha;-(D)-glucosyl-&alpha;-(L)-oxyisovaléryl)-SMS

[&alpha;]D20 = -44,3 (c = 1, dans AcOH à 95%) F : 1,00
Exemple 29 [N-acétylmuramyl-(D)Phe]-SMS

[&alpha;]D20 = -15,4 (c = 0,13, dans AcOH à 95%) F : 0,9
Exemple 30
ss-D-Glucosyl-thiocarbamoyl-SMS

Example 27 N &α; - (O-ss- (D) -glucosyloxyisobutyryl) -SMS

[&alpha;] D20 = -32.9 (c = 1, in 95% AcOH) F: 0.93
Example 28
N &alpha;-( O- &alpha;-( D) -glucosyl- &alpha;-( L) -oxyisovaléryl) -SMS

[&alpha;] D20 = -44.3 (c = 1, in 95% AcOH) F: 1.00
Example 29 [N-acetylmuramyl- (D) Phe] -SMS

[&alpha;] D20 = -15.4 (c = 0.13, in 95% AcOH) F: 0.9
Example 30
ss-D-Glucosyl-thiocarbamoyl-SMS

A 620 mg of # -Fmoc-SMS in 50 ml of
CH 3 CN / H 2 O 3: 1, 0.45 ml of triethylamine and then 272 mg of 2,3,4,6-tetra-O-acetyl-β-D-glucosylisothiocyanate are added and the mixture is stirred for one hour at Room temperature.

After evaporation of the mixture in vacuo, the residue is taken up in a little methanol and treated with diisopropyl ether, which causes the product to precipitate in practically pure form.

To remove the Fmoc group and the acetyl groups, the product is treated in 50 ml of absolute methanol with a catalytic amount of 1N NaOCH3 in methanol. After 30 minutes (verification by thin layer chromatography), the mixture is neutralized with 1% acetic acid and evaporated under vacuum. The residue is taken up in water and extracted with ethyl acetate. The aqueous phase is lyophilized and the residue is purified on silica gel and demineralized for example on
Duolite. The title compound is thus obtained in the form of a lyophilizate.

 [&alpha;] D20 = -48.5 (c = 1, 95% AcOH) F: 1.

The - Fmoc-SMS used as a starting product can be prepared as follows
To 5 g of SMS acetate and 5 g of NaHCO 3 in 100 ml of a 3: 1 mixture of DMF and water, 1.6 g of Fmoc-HOSu are added. After one hour at room temperature, the mixture is diluted with 400 ml of water and extracted with 250 ml of a 95: 5 mixture of ethyl acetate and methanol. The organic phase is dried over Na 2 SO 4 and concentrated. After chromatography on silica gel, the product is obtained in the form of an amorphous substance.

 [&alpha;] D20 = 24.3 (c = 1.13 in DMF).

By proceeding in a manner analogous to that described in Example 30, the following compounds can be obtained:
Example 31
Cellobiosylthiocarbamoyl-SMS

[&alpha;] D20: -4.3 (c = 1, in 95% AcOH) F = 0.87
As the starting material, octaacetyl-cellobiosyl isothiocyanate is used.

[&alpha;]D20: = 39,9 (c = 1, dans AcOH à 95%) F = 0.81
Comme produit de depart, on utilise l'isocyanate de 2,3,4,6-tétra-O-acétyl-ss-D-glucosyle.Example 32
ss-D-glucosylcarbamoyl-SMS

[&alpha;] D20: = 39.9 (c = 1, in 95% AcOH) F = 0.81
The starting material used is 2,3,4,6-tetra-O-acetyl-ss-D-glucosyl isocyanate.

[&alpha;]D20 = -37,9 (c = 1, dans AcOH à 95%) F = 0,85
Comme produit de départ, on utilise l'iso-
cyanate d'octa-acétyl-cellobiosyle. Example 33
Cellobiosylcarboamoyl-SMS

[&alpha;] D20 = -37.9 (c = 1, in 95% AcOH) F = 0.85
As a starting material, the iso-
octaacetyl-cellobiosyl cyanate.

Example 34
1-deoxy-D-sorbityl-SMS

0.5 mg of the title compound of Example 1 in 50 ml of methanol is treated first with
NaBH4 then with 5% acetic acid, under conditions such that the pH does not exceed 7. The total amount of NaBH4 used is about 10 equivalents
After the reaction was complete (4-5 hours), the mixture was treated with acetic acid to destroy excess NaBH 4 and the mixture was concentrated in vacuo.

The residue is demineralized for example on duolite and purified on silica gel. The main product is the title compound in the form of a lyophilizate, accompanied by 1-deoxy-D-mannityl-SMS.

 [&alpha;] D20 = -17.6 (c = 1 in 95% AcOH) F = 0.88.

[&alpha;]D20 = +1,6 (c = 1 dans AcOH) F:0,9.Example 35 α-D-glucosyl (1-4) deoxysorbityl-SMS
By proceeding in a similar manner to that described in Example 34 and using as starting material the title compound of Example 2, the following compound is obtained:

[&alpha;] D20 = +1.6 (c = 1 in AcOH) F: 0.9.

Example 36) 1,2-dideoxy-sorbityl-SMS

0.55 g of Boc-SMS are treated in 30 ml of a 3: 7 mixture of dioxane and water per 50 ml of
NaBH3CN. 250 mg of 2-deoxy-D-glucose are added, the pH of the mixture is adjusted to 7 with 0.1 N HCl and heated at 100 for 6 hours. The mixture is cooled, frozen and lyophilized. The residue is taken up in ethyl acetate (50 ml) and extracted with water. The organic phase is dried and evaporated under vacuum. The BOC group is removed according to conventional methods using TFA. The product is purified on silica gel and demineralized for example on dulite, which gives the title compound. [&alpha;] D20 = 25.5 (c = 1, 95% AcOH) F: 0.83.

By proceeding analogously, the compound of Example 34 above (starting from glucose) and the compound of Example 35 above (starting from maltose) can be prepared.
Example 37 N-alpha-isocapropyl-des (1-4) - [Ala7, NE- (1-deoxyfructosyl) -Lys11,18] calcitonin of saunon

 10.3 g of N-alpha-iso-caproyl-des (1-4) - [Ala7] -calcitonin polyacetate of salmon and 1.8 g of D (+) - glucose are dissolved in a mixture of 94 ml of DMF. and 6 ml of acetic acid. After 2 hours at 500, the product is completely precipitated by addition of ether, filtered off, washed with ether and dried under vacuum. To purify the product, about 5 to 10 g of produced in water, the solution was injected onto a 4 x 25 cm reverse phase column (silica gel, C-18) and chromatographed with a gradient of water and from 0 to 80% of a solvent mixture. comprising 38 parts of water, 60 parts of acetonitrile and 2 parts of 85% phosphoric acid.

The fractions containing the pure product are combined, filtered through a column of about 100 ml of a slightly basic ion exchange resin in acetate form, and washed with water. The filtrate is lyophilized to give the title compound as polyacetate, polyhydrate.

 [?] OD = -34.8 (c = 0.73 in 95% CH3COOH) F: 0.93.

Fast atom bombardment mass spectrum: 3407 (MH +).

N-alpha-isocapyroyl-Ser-Thr-Ala-Val-Leu-Gly-Lys-
Leu-Gln-Ser-Glu-Leu-His-Lys-Leu-Gln-Thr-Thr-Pro-Arg-Thr-Asn
Thr-Gly-Ser-Gly-Thr-Pro-NH2 used as starting material can be prepared as follows: a) N-alpha-isocaproyl-Ser (But) -Thr (But) -Ala-Val-Leu-OCH2-phenyl-
(P) OCH2-co (polystyrene-1% divinylbenzene)
1 g of p-hydroxymethyl is allowed to swell
phenoxymethyl-co (polystyrene-1% divinylbenzene)
in a 1: 4 (v / v) mixture of DMF and
methylene chloride, filtered off and mixed with a solution of 0.74 g of Fmoc-leucine and 0.19 g of 1-hydroxybenzotriazole in 5 ml of the solvent mixture mentioned above.While stirring, add 0, 43 g of dicyclohexylcarbodiimide and 85 mg of 4-dimethylaminopyridine each in 5 ml of the same solvent mixture. The mixture is stirred for 16 hours at 200, filtered and washed with the solvent mixture and then with dimethylformamide. This gives the Fmoc-Leu
OCH2-phenyl- (p) OCH2-co (polystyrene-1% divinylbenzene).

The Na-Fmoc group of this product (1.56 g corresponding to 0.7 mMol) is removed by treatment with pyridine (20% vol / vol) in
DMF for 10 minutes. Wash well with
DMF and then add 0.71 g of Fmoc-Val-OH, 0.28 g of 1-hydroxybenzotriazole and 0.32 ml of diisopropylcarbodiimide, each product being dissolved in 5 ml of DMF. After 45 minutes, the mixture is filtered off and the resin rinsed well with DMF. The elimination of the Na group is repeated. Fmoc and the coupling with the following amino acids in the order indicated
Fmoc-Ala-OH (0.65 g), Fmoc-Thr (But) -OH (0.83 g) and Fmoc-Ser (But) -OH (0.80 g). In the last reaction cycle (elimination of the Fmoc protecting group, acylation with the protected amino acid), the amino acid derivative is replaced by isocaproic acid (0.41 g), the amount of 1-hydroxybenzotriazole is increased to 0.53 g and that of diisopropylcarbodiimide at 0.54 g, and the coupling is carried out for 15 hours. The resin with the protected peptide is washed well with DMF and methylene chloride and
dried under vacuum at 400C for 15 hours, which
gives the resin with the protected peptide in form
an uncoloured powder.

b) N-alpha-isocapropyl-Ser-Thr-Ala-Val-Leu-OH
1 g of the title compound of the example is stirred
a) above, in a mixture of 5 ml of trifluoro acid
roacetic acid and 5 ml of methylene chloride. We
filter the product, wash it with 5 ml of the same
mixture, then with methylene chloride,
strongly concentrated under vacuum and rushed
entirely by addition of ether. We wash well
precipitated with ether and dried under vacuum on
solid potassium hydroxide, which gives the
compound of the title in the form of an amorphous powder
colorless.

c) N &alpha; -isocaproyl-Ser-Thr-Ala-Val-Leu-Gly-Lys (Boc) -Leu-Ser-
Gln-Glu (OBu) Leu-His-Lys (Boc) -Leu-Gln-Thr-Tyr-Pro-Arg
Thr-Asn-Thr-Gly-Gly-Ser-Thr-Pro-NH2
To a solution of 0.165 g of the
step b) above in 7 ml of DMF, one adds
0.59 g of H-Gly-Lys (Boc) -Leu-Ser hydrochloride
Gln-Glu (OBu) Leu-His-Lys (Boc) -Leu-Gln-Thr-Tyr-Pro
Arg-Thr-Asn-Thr-Gly-Ser-Gly-Thr-Pro-NH2, 0.017 g
3,4-dihydro-3-hydroxy-4-oxo-1,2,3-benzotriazine,
0.065 g of dicyclohexylcarbodiimide and sufficiently
of N-ethyl-N, N-diisopropylamine for a sample
lon of the reaction mixture indicates a pH of about
6 on moist pH paper. After 16 hours, we
precipitates the product by addition of ether
and dry it. This gives the compound of the
title.

d) N &alpha; -isocaproyl-Ser-Thr-Ala-Val-Leu-Gly-Lys-Leu-Ser-Gln-Glu-
Leu-His-Lys-Leu-Gln-Thr-Tyr-Arg-Pro-Thr-Asn-Thr-Gly-Ser
Gly-Thr-Pro-NH2
0.50 g of the peptide is partially dissolved
protected from step c) above in a mixture
of 50% trifluoroacetic acid (vol./vol.) and
methylene chloride. After 1 hour, we add
50 ml of ether containing 0.6 mMol of HCl. Filter
the product is washed with ether and dried
under vacuum. The product is purified by chromato
reversed phase graphy with a gradient of aces
tonitrile in H3P04 (2%). Fractions together
containing the substance in pure form are filtered
on basic ion exchange resin under
acetate form, and the filtrate is lyophilized,
which gives the title compound in the form of poly
polyhydrated acetate.

 [&alpha;] D20 = -32.2 (c = 0.3 in 95% AcOH) F: 0.87.

Example 38 N α -isocaproyl-des- (1-4) - [Ala7, N # (α-D-glucosyl (1-4) -das-oxyfructosyl) -Lys [1,18] salmon calcitonin

 The corresponding di-N-malulosyl derivative is prepared analogously to that described in Example 37 using D (+) maltose monohydrate in place of D (+) - glucose. The reaction time at 50 C is 15 hours. The isolation and purification of the product are identical. The title compound is obtained in the form of polyacetate polyhydrate.

Fast atom bombardment mass spectrum: 3730.9 (MH +).

 [&alpha;] D20 = -15.2 (c = 0.16 in 95% AcOH), F: 0.97.

[&alpha;]D20= -40,0 (c = 0,27 dans AcOH à 95%) F : 0,84.By proceeding in a manner analogous to that described in Example 37, the following compounds can be prepared:
Example 39 Nazi-α-isocaproyl- [N- (1-deoxyfructosyl) -Lys7] -calcitonin of salmon- (5-32) amide

[&alpha;] D20 = -40.0 (c = 0.27 in 95% AcOH) F: 0.84.

Example 40 N &alpha; Lys-N #, Lys18N # -tris- (1-deoxyfructosyl) -calcitonin of salmon

[&alpha;]D20 =-5,3 (c = 0,38 dans AcOH à 95%) F:0,75.

[&alpha;] D20 = -5.3 (c = 0.38 in 95% AcOH) F: 0.75.

Example 41
N-alpha-isocaproyl-des (1-4) - [N # - (1-deoxyfructosyl) -Lys7,18] calcitonin from salmon- (5-32) amide

[&alpha;] D20 = - 37.4 (c = 0.155, AcOH, 95%) F = 0.84
Example 42 N α -quinoyl- [Ala 7] -calcitonin from salmon- (5-32) -amide

 The title compound is prepared analogously to that described in Example 21.

 [&alpha;] D20 -35.7 (c = 0.37 in 95% AcOH) F: 0.88.

Example 43 Salmon- (4-32) -ammonium-quinoyl- [Ala7] -calcitonin

 The title compound is prepared analogously to that described in Example 21.

 [&alpha;] D20 = -39.3 (c = 0.29 in 95% AcOH) F: 0.89.

 [Ala7] -calcitonin of salmon - [5-32] amide and the [Ala7] -calcitonin of salmon- (4-32) -amide necessary as starting materials for Examples 42 and 43 can be prepared analogously. to that described for the starting material of Example 37.

Example 44 [N-Deoxy-Fructosyl-Cys] -Oxytocin

0.5 g of D (+) - glucose and 0.5 g of oxytocin are dissolved in 50 ml of a 9/1 mixture of MeOH and
AcOH and the mixture is maintained for 3 hours at 65.

The solution is then concentrated by evaporation, chromatographed on silica gel and the salt is freed from Duolite (gradient: H 2 O / ethanol / AcOH). A white lyophilizate is obtained.

 [&alpha;] D20 = -23 (c = 0.32 in 95% AcOH) F: 0.95.

Example 45
Ac- (D) -Phe (4-Cl) - (D) Phe (4-Cl) - (D) Trp-Ser-Tyr

60 mg of Ac- (D) -Phe- (4-Cl) - (D) are dissolved
Phe (4-Cl) - (D) -Trp-Ser-Tyr- (D) -Lys-Leu-Arg-Pro- (D) -Ala
NH2 and 72 mg of D (+) glucose in a mixture of 10 ml
of MeOH and 1 ml of AcOH and the mixture is stirred for about 20 hours at 600. The product is precipitated by
addition of ether and centrifugation. Dissolve
residue in about 100 ml of H 2 O and adjust the pH
to 8 with a diluted solution of NaOH. We passed
the product on a column of Duolite ES 861 eluting with a gradient H2O # dioxane-H2O-AcOH
(60: 40: 3). Fractions containing the desired product
are concentrated under vacuum and lyophilized. We obtain
thus the title compound.

 [&alpha;] D20 = -25 (c = 0.5 in 95% AcOH) F: 0.83.

Example 46
N &alpha; Al, N &α;ss1; N # B29-tris (1-deoxyfructosyl) -insulin of proc
It is stirred for 2 hours at 60 a suspen
1 g (0.17 mmol) of pork insulin and 0.47 g
(2.6 mmol) glucose in 10 ml of a 9: 1 mixture
DMF and acetic acid. Solvent is removed
by evaporation at 30 ° C. under high vacuum, the residue is dissolved in 300 ml of water, the pH is adjusted to 7 and the solution is passed through a demineralising column.
(Duolite ES 861, 2.5 x 15 cm). Glucose is eluted with water and the peptide with a 59: 39: 2 mixture of isopropanol, water and glacial acetic acid.

After evaporation of the solvent and lyophilization, the solution is dissolved. substance in 300 ml of water and purified by reverse phase chromatography (2 x 25 cm
RP 18, 10 nm, buffer solution: 57 mmol NaClO4, 20 mmol triethylamine, 8.4 mmol phosphoric acid, pH set to 3 with 4N NaOH; gradient: 0-65% AB:
Buffer A: Buffer pH 3 / acetonitrile 9: 1
Buffer B: Buffer pH 3 / acetonitrile 4: 6).

 The fractions containing the title compound are collected, pooled, concentrated, diluted with 300 ml of water and passed through a demineralising column as described above. The salts are eluted with water and the peptide is eluted with a 59: 39: 2 mixture of isopropanol, water and glacial acetic acid.

The appropriate fractions are combined, concentrated and lyophilized to give the title compound.

 = -56.3 (c = 0.5 in 95% AcOH) F: 0.88.

@@@@
Example 47 N &alpha; Lys, Lys-N # -tris- (1-deoxyfructosyl)
(D [Ala] -hpGRF- (1-29) -NYH2

 By proceeding in a manner analogous to that described in Example 37 and using the [DAla] hpGRF as starting product 2, the title compound is obtained.

 [&alpha;] D20 = -5.6 (c = 0.2 in 95% AcOH) F: 0.82.

Example 48 N &alpha; N # -bis (1-deoxyfructosyl) -Lys-vasopressin

 A suspension of 118 mg (0.1 mmol) of Lys8-vasopressin and 360 mg (2 mmol) of glucose in 5 ml of a 9: 1 mixture of methanol and acetic acid is stirred for 2 to 4 hours. glacial. The solvent is removed in vacuo, the residue is taken up in 30 ml of water and the solution is lyophilized. To remove excess glucose, the peptide (solution in 40 ml of water, pH 7.3) is adsorbed on a column of Duolite (1.5 x 10 cm). The glucose is eluted with water and the peptide with a mixture of isopropanol, water and glacial acetic acid (59: 39: 2). After evaporation under vacuum and lyophilization, the product is purified by chromatography on silica gel. (chloroform / methanol / glacial acetic acid / water 7: 4: 1: 1).

 The fractions containing the pure substance are pooled, concentrated and lyophilized. The title compound is thus obtained.

 [&alpha;] D20 = -52 (c = 0.5 in 95% AcOH) F: 0.84.

Example 49

By proceeding in a manner analogous to that described in Example 45 and using as starting material the Ac- (D) -Phe (pCl) - (D) Phe (pCl) - (D) -Trp-Ser-
Lys- (D) -Phe-Leu-Arg-Pro (D) Ala-NH 2 acetate and the
D (+) glucose gives the title compound.

 [&alpha;] D20 = -36 (c = 0.5 in 95% AcOH) F = 0.86.

(D)Pre(pCl)-(D)Phe(pCl)-(D)Trp-Ser-Tyr-(D)-Phe-Leu-Arg-Pre (D)Ala-NH2, Acétate
En procédant de manière analogue à celle décrite à l'exemple 45 et en utilisant comme produit de départ le H-(D)-Phe)pCl)-(D)Phe(pCl)-(D)-Trp-Ser-
Tyr-(D)-Phe-Leu-Arg-Pro-(D)Ala-NHY2 acétate et le
D(+)glucose, on obtient le composé du titre [&alpha;]D20 = -32 (c = 0,5 dans AcOH à 95%) F:0,94.Example 50

(D) Pre (pCl) - (D) Phe (pCl) - (D) Trp-Ser-Tyr- (D) -Phe-Leu-Arg-Pre (D) Ala-NH 2, Acetate
By proceeding in a manner analogous to that described in Example 45 and using as starting material the H- (D) -Phe) pCl) - (D) Phe (pCl) - (D) -Trp-Ser-
Tyr- (D) -Phe-Leu-Arg-Pro- (D) Ala-NHY2 acetate and the
D (+) glucose gives the title compound [& alpha] D20 = -32 (c = 0.5 in 95% AcOH) F: 0.94.

Example 51

 200 mg of H- (D) Phe (pCl) - (D) Phe (pCl) - (D) Trp-Ser-Tyr- (D) Lys-Leu-Arg-Pro- (200 mg) are stirred for about 3 hours ( D) Ala-NH2 and 520 mg of D (+) glucose in a 15: 1 mixture of DMF and AcOH. The mixture is concentrated in vacuo, precipitated with ether, filtered and the product dried.

The residue is purified as follows 1) Adsorption on Duolite ES 861 and solution with a
dioxane / H 2 O / AcOH mixture.

2) silica gel column chromatography
(CHCl3 / AcOH / H2O mixture).

3) reverse phase preparative chromatography (HPLC)
(C18-silica gel). Elution with acetonitrile gradient in 2% E3PO4.

Fractions containing the title compound are pooled, filtered on a column containing a weakly basic ion exchange resin in acetate form, concentrated and lyophilized.
This gives the title compound.

 [&alpha;] D20 = -22.6 (c = 0.5 in 95% AcOH) F: 0.63.

The following example illustrates the solid phase process for the preparation of the peptide-alcohol

Example 52 1) N-CF3CO-threoninol acetal of p-formylphenoxy acid
acetic
To 200 ml of methanol, add under running
nitrogen 105 g of L-threoninol. To the solution
limpid thus obtained, it is added drop by drop
to 0 a solution of 200 ml of trifluoroacetate
of methyl in 250 ml of methanol. The mixture is now
held at a temperature of about 10 by cooling with an ice bath. After an hour and a half, free threoninol can no longer be detected in the mixture.

By evaporation at 400, a white crystallized residue is obtained. The residue is dissolved in ethyl acetate (200 ml) and precipitated by the addition of 100 ml hexane. The mixture is cooled to 0, the product is filtered, washed with hexane and dried at room temperature. This gives N-trifluoroacetylthreoninol
Under nitrogen, 50.3 g (0.25 mol) of the product obtained above are dissolved in 1.25 liters of tetrahydrofuran and 75 ml of trimethylchlorosilane are added dropwise at room temperature. Immediately afterwards, a mixture of 70 ml of triethylamine and 250 ml of tetrahydrofuran is added. A white suspension is formed which is stirred for 4 hours at room temperature. The mixture is then filtered and the filtrate is evaporated to about 40 °. which gives an oil.

 The oil is dissolved in 1.5 liters of methylene chloride and 90.4 g of p-formylphenoxyacetic acid are added in portions. 9 ml of trimethylsilyl trifluoromethanesulfonate are then added in portions, the mixture is stirred for 24 hours at room temperature, filtered and the residue is washed well with methylene chloride.

The filtrate is evaporated to 400, giving an orange-red resinous product. This product is chromatographed on silica gel using ethyl acetate as eluent. By evaporation
appropriate fractions, we get the compound
of the title having a purity of 97% (HPLC).

2) Synthesis of octa-eeetlde ~ erotege
17.2 g of aminomethylated polystyrene (0.7%
by weight of nitrogen corresponding to 0.5 mmole of
amino-methyl groups per g of resin) are
suspension in 80 ml of a 4: 1 mixture of chloride
of methylene and DMF.and successively added
4.17 g of the final product of Step 1, 1.6 g
of hydroxybenzotriazole and 4.0 g of DCCI. After
2 hours stirring at room temperature,
the Kaiser test is negative. Then filter
mix and wash. We put again the resin
in suspension in 100 ml of a 3: 1 mixture of
tetrahydrofuran and methanol and added by
portions 10.4 g of sodium borohydride. We shake
the mixture for 6 hours at room temperature,
it is filtered and the resin washed. We put again
the resin suspended in a 4: 1 mixture of chlo
of methylene and DMF and 5.57 g of
Fmoc-Cys (St-Bu) OH, 1.74 g of hydroxybenzotriazole
and 3.6 g of DCCI. After elimination of the group
Fmoc protector with piperidine (2 x 20 minutes),
we couple successively in the order indicated
following amino acids protected with nitrogen by
a Fmoc group using hydroxybenzoate
triazole and DCCI: ThrOH, Lys (BOC) -OH,
D-TrpOH, Phe-OH, Cys (S-tBu) OH and D-Phe-OH. We
thus obtain the Fmoc-octapeptide on resin.

 Final charge = 0.26 mmol / g.

3) Oxidation and split
The resin thus obtained is suspended
in 100 ml of a 1: 1 mixture of trifluoro
ethanol and methylene chloride. We add
then 50 ml of tributylphosphine and stirred
mixing for 70 hours at the temperature
room. The mixture is filtered, washed and
100 ml of a 1: 1 mixture of tetrahydro
furan and a 1N solution of ammonium acetate.

1.1 ml of an aqueous solution is then added
30% oxygenated water and stir the mixture to
room temperature for 24 hours. We
then add a mixture of 20 ml of tri acid
fluoroacetic acid, 80 ml of methylene chloride,
10 ml of water and 2 ml of thioanisole and
Stir for 2 hours. After filtration, wash
with trifluoroacetic acid and
methylene chloride. 200 ml of ether are added
diethyl chloride to the filtrate, filtered off
the product that has precipitated, the product is dissolved
in an aqueous buffer, the solution is demineralized
by treatment with Duolite and freeze-dried
in the form of acetate. The octa-peptide is thus obtained
in the form of acetate.

The compounds of the preceding examples, for
example the compounds of Examples 1 and 2 can
be prepared in a similar way.

Example 53
Preparation of N- [alpha] -glucosyl (1-4) -deoxyfructosyl)
SMS (see example 2)
Using the procedure described in Example 52, 393 g of the octapeptide bound to the resin are prepared.

The protecting groups of cysteine are removed by reduction. The peptide bound to the resin is oxidized to cyclic octapeptide by the oxygenated water in a mixture of tetrahydrofuran and water.

After washing in tetrahydrofuran and then in DMF, the resin is extracted with 3600 ml of an 8: 1 mixture of DMF and
AcOH. The suspension is treated with 526 g of D (+) maltose monohydrate. The mixture is heated to 600 and stirred for 18 hours at this temperature.

 The mixture is cooled and the resin is separated by filtration and washed successively with DMF and methanol and then with methylene chloride. The peptide is cleaved from the resin for 1 hour with a mixture of 2900 ml of methylene chloride and 716 ml of trifluoroacetic acid with a trace of water.

 The filtrate is then stirred, 597 g of sodium carbonate are added in portions, stirred for 30 minutes and filtered. The residue is washed with methylene chloride and methanol and the filtrate is concentrated to dryness. It is demineralized using a column of polystyrene of the Duolite type, or in the reverse phase liquid phase (HPLC), for example on silica gel treated with silicone and comprising long-chain fatty alcohol groups (for example Labomatic , Switzerland, mark HB-SIL-18-20-100). The title compound is thus obtained in the pure state.

 The compounds of the invention have interesting pharmacological properties and can therefore be used therapeutically as medicaments.

 The activity of the compounds of the invention can be demonstrated in conventional biopharmaceutical and pharmaceutical assays. The compounds are generally at least as potent as the unmodified peptide (ie the corresponding non-glucosylated peptide) after administration by injection or orally. They are generally better absorbed, are more easily soluble in water, and have a longer duration of action.

 The compounds of the invention are therefore indicated to be used for the same indications as those of the unmodified peptides.

 The compounds of the invention can be compared to unmodified peptides in conventional bioavailability assays.

 Compounds of the invention may, for example, be detected in blood plasma for a longer period after administration than unmodified peptides, as is apparent from conventional bioavailability assays.

 The compounds of the invention and the unmodified peptide may be administered orally or intravenously, for example to dogs, in a single dose sufficient to achieve the desired therapeutic effect.

 The doses used are those for detecting the peptide or one of its metabolites in the blood. The detection can be carried out according to the usual methods, for example according to the radioimmunoassay (RIA) method.

 In the test mentioned above, it has been found, for example, that the compound of Example 2 administered orally produces blood concentrations ten times higher than those obtained with octreotide.

 The absolute bioavailability of the compound of example 2 administered orally or intravenously, determined by the AUC (area under the curve) is 5 times greater than that of octreotide.

The half-life of intravenous elimination is approximately 2.3 hours whereas it is approximately 0.5 hours for octreotide.

 In addition, the compounds of the invention have the advantage of being eliminated largely by the kidneys. This can be demonstrated by conventional tests.

 Male rats (weighing 225-375 g) are fasted and given oral water (50 ml / kg). The animals are anesthetized after 30 minutes with, for example, inactin (100 mg / kg intraperitoneally). A cannula is placed in the bile duct and bladder. The two jugular veins are exposed. In one of them, a 5% glucose and 1% ethanol infusion (5 ml / hour) is used to stimulate diuresis. The other vein is used to take blood samples (0.5 ml) every hour for 4 hours.

 The compound of the invention and the unmodified peptide are administered subcutaneously at a dose of from about 10 to about 1000 micrograms / kg. The concentration of the compound is determined according to the usual methods, for example by RIA.

In the above test, for example, the following results were obtained with the compound of Example 2 and Octreotide at 10 microgram / kg:
Percentage eliminated by lane
Urinary Biliary
Compound of Example 2 1.6 36
Octreotide 22 19
While octreotide is eliminated by the bile and urinary routes, the compound of Example 2 is essentially eliminated urinary.

Improved absorption of the compounds of the invention can be shown by oral administration, as follows:
The compound of the invention and the unmodified peptide are orally administered to OFA rats (eg 10 mg / kg). At various precise time intervals, for example 15, 30 and 60 minutes, blood samples are taken and their active substance content is analyzed, for example by RIA.

In this test, it has been found, for example, that the compound of Example 44, administered at a dose of 10 mg / kg, has an absorption 50 to 100% higher than that of the unmodified peptide, oxytocin. The results are as follows:
Plasma levels in the rat following administration by
oral route. The results are given in ng / ml
15 minutes 30 minutes 60 minutes
Oxytocin 7.53 3.60 2.55
Composed from example 44 11.79 6.98 3.98
The pharmacological activities of the compounds of the invention can be demonstrated in usual pharmacological tests, for example after injection, and, if necessary, compared with those of the unmodified peptides, for example in terms of potency and duration of administration. action.

 For example, it is possible to carry out pharmacological tests to verify in animals the effects of the compounds of the invention on hormones. Thus, compounds that inhibit the secretion of hormones can be tested to determine the decline in blood levels of the hormone.

Compounds of the invention which inhibit the secretion of growth hormone, in particular compounds of the formula
VIII, and more particularly the compounds of formula VIIIa to f, and reduce growth hormone concentrations in the blood, may be subjected to the following tests:
Rhesus monkeys (at least 5 monkeys) are fasted on chairs and administered the compound of the invention in a piece of banana serving as a vehicle. The compounds are administered at a dose of from about 0.1 mg / kg to about 10 mg / kg orally. Blood from the saphenous vein is removed by a catheter. The concentration of growth hormone in the blood is measured by RIA.

 In this test carried out on rhesus monkeys, it has been found, for example, that the compound of Example 2 administered at a dose of 0.1 mg / kg causes a decrease in the secretion of growth hormone of about 50%. for more than 10 hours, whereas this effect lasts only 5 hours with the unmodified peptide, octreotide.

 In another test, male rats were decapitated and blood taken 1 hour after logarithmic administration of the growth hormone secretory inhibiting compound. The growth hormone level in the blood is determined by RIA. In this test, the compounds of the invention are active at doses of from about 0.02 to about 30 micrograms / kg subcutaneously. It has been found, for example, that the compounds of Examples 1, 2, 21 and 24 administered subcutaneously, have an ID50 of 0.045, 0.190, 0.3 and 0.2 micrograms / kg, respectively, the DI50 of natural stomatostatin administered subcutaneously in the dermal test being 93 micrograms / kg. (The ID50 indicates the amount of compound required to lower the growth hormone content by 50% over untreated control animals).

 In contrast to natural somatostin, the growth hormone secretory inhibiting compounds of the invention are highly active in this assay for a long time (e.g. 6 hours).

The inhibitory activity of these compounds was also demonstrated after oral administration to male rats with estradiol implants. In this test, relatively small variations in the level of growth hormone are observed. The test is performed as follows:
To male rats with a weight of about 300 g, a tube (length = 50 mm, diameter = 3 mm) of silastic with 50 mg of estradiol is implanted under the dorsal skin, under anesthesia with ether. These animals are used several times (1-6 months later) for testing. The test substance is administered subcutaneously or orally.

 Just before, and at various time intervals after the administration of the substance, about 0.8 ml of retro-orbital plexus blood is taken, centrifuged and the growth hormone level is determined. serum by RIA.

The compounds of the invention are more active after oral administration than the corresponding unmodified peptides, even after several hours. The ID50 of each compound of Examples 1 and 2 is, after 2 hours, about 17 to 40 times lower than that of the unmodified peptide, octreotide. Other results are:
Compound of the example DI50 orally microgram / kg 21 500 24 25
Octreotide 1400
The compounds of the invention are therefore indicated when an inhibition of the secretion of growth hormone is necessary. Indications include diabetes mellitus, prevention and treatment of angiopathy and proliferative retinopathy, as well as acromegaly.

 The inhibitory action of the compounds of the invention on the secretion of growth hormone also inhibits pancreatic secretion. This inhibition can be demonstrated in animal tests. The method described in Scand. J.

Gastroint. 6, 423 (1975) by S.J. Konturek et al. can be used.

 Compounds of the invention that inhibit the secretion of growth hormone also inhibit secretion of gastric juice and increase the pH of gastric juice by several units.

This activity of the compounds of the invention can be demonstrated for example in the following test:
Compounds of the invention which inhibit the secretion of growth hormone at a dose of from about 0.05 mg / kg to about 5 mg / kg are administered to fasting rats in which a fistula has been performed. in the stomach. After 1 hour, the fistula is opened and the gastric juice is removed every 30 minutes. The volumes collected are recorded and the acid concentration is determined.

 In this test, the compound of Example 2 causes a pH increase of 6 to 8 for 3.5 hours. Octreotide caused the pH values to increase from 6 to 7 for only 2 hours. This test shows that the compound of Example 2 is at least 60 times more active than cimetidine.

The compounds of the invention which inhibit the secretion of growth hormone, in particular compounds of the formula
VIII, can therefore be used therapeutically for the treatment of gastrointestinal disorders, for example for gastric ulcers, gastrointestinal bleeding, acute pancreatitis and gastroenteropancreatic tumors (for example, vipomas insulinomas, glucagonomers etc.).

 The compounds of the invention which inhibit the secretion of growth hormone also inhibit the proliferation and / or keratinization of epidermal cells, and are therefore indicated for the treatment of dermatological diseases associated with pathological proliferation and / or keratinization. epidermal cells, especially for the treatment of psoriasis.

 In addition, these compounds may also be used therapeutically in the treatment of degenerative senile dementia, also senile dementia of the Alzheimer type (DSTA), or in the treatment of vascular pain of the face (repetitive headaches).

 For all these indications, the compounds of the invention which inhibit the secretion of growth hormone are advantageously administered at doses of between 2 micrograms and 20 milligrams. If necessary, the compounds may be administered in divided doses 2-4 times daily as unit doses, or in a sustained release form.

The unit doses can comprise between 0.5 microgram and 10 mg of active substance.

The glucosyl derivatives of calcitonin and its derivatives and analogues according to the invention, more particularly the derivatives of formula X, reduce the calcium levels in the blood plasma. In addition, they act as functional antagonists of parathyroid hormone to give a positive calcium balance in favor of the bone. The hypocalcemic activity of the new compounds can be determined according to the usual methods, for example according to the method of M. Azria et al., Exhibited on 2-4 October 1984 during the
Symposium on calcitonin held in Milan and published as a short paper in "Current Clinical Practice Series"
No. 42, Excerpta Medica 1986, p. 104. In this method, a calcium ion selective electrode is used to continuously determine the calcium ion content in the blood of young rabbits or dogs. The compounds are administered by intravenous route from about 0.1 to about 10 micrograms / kg, for example corresponding to about 1 international unit per kg.

The measurements are carried out for 5 hours and the AUCs are calculated.

 Other assays may also be performed with the compounds of the invention, for example the assay described by M. Kumar et al., J. Endocrinology (1965), p. 469, and practiced on rats administered different doses causing hypocalcemia, ie from 300 to 6000 IU per mg for the compounds of the invention having hypocalcemic activity.

 It has been found, for example, that the compounds of Examples 37 and 38 administered intravenously to dogs (5 μg / kg) have a much longer duration of action than the unmodified peptide.

In this test, a decrease in blood calcium level of between 15 and 18% for compounds 37 and 38 is observed after 3 hours; after 6 hours, no blood loss can be detected in the blood for the unmodified peptide, whereas with the compounds of Examples 37 and 38, the decrease in calcium level was still as pronounced as after 3 hours.

 The hypocalcemic compounds of the invention can therefore be used therapeutically for the treatment of all the conditions requiring a drop in the level of calcium in the blood plasma or an effect on bone metabolism, for example for the treatment of hypercalcemia resulting from endogenous thyrocalcitonin deficiency due to loss of thyroid C cell secretory function or hyperfunction of the parathyroid gland.They can also be used for the treatment of all bone conditions associated with increased bone friability or which fixation of calcium in the bone is necessary, for example for the treatment of osteoporosis of various origin (for example post-climacteric, post-traumatic, following corticotherapy or immobilization, of malignant origin etc.) ), fractures, osteomalacia, renal osteodystrophy caused by rickets, pain, for example, bone pain associated with osteoporosis, neurodystrophic disorders, Paget's disease and for combination therapy with calcium or phosphates.

For the above calcium-related indications, the appropriate daily dose is between about 5 and about 1500
IU, administered daily at once or, if necessary, every 2 or 3 days.

 The compounds of the invention, which are glucosylated derivatives of LHRH or its analogs, inhibit the secretion of luteinizing hormone, for example as indicated by inhibition of ovulation in animals. This test is carried out according to the method described by M. Marko and E. Flückiger, in Experientia 30, p. 1174-1176 (1974).

In general, these compounds of the invention are active at a dose of from about 0.0005 to about 10 mg / kg. For example, the compound of Example 45 administered subcutaneously is active at a dose of 0.01 mg / kg. The inhibitory effect of the compounds on the secretion of luteinizing hormone can also be determined in vitro. Pituitary cell cultures are prepared according to the method described by Vale (W. Vale and G. Grant,
Enzymology 37, p. 82-93; 1975) and by Mr. Marko and D. Rimer in Life
Science, 33, p. 233-240 (1983). The primary cells are kept for 4 days in an incubator at 37 ° C. The cells are then washed and incubated for 3 hours in 1 ml of medium containing LHRH or the test compound. At the end of incubation the supernatant is recovered and the activity of luteinizing hormone is determined using the radioimmunoassay (RIA) method.

 In this test, the compounds of the invention are generally active at a concentration of about 10-12 to about 10 -7, and inhibit LHRH-induced luteinizing hormone secretion as a function of dose.

 For all indications of LHRH, the appropriate daily dose is between 2 micrograms and 20 mg of active substance. If necessary, the compounds may be administered in divided doses 2-4 times daily as unit doses or, if necessary, in sustained release form. Such unit doses comprise between about 0.5 micrograms and 10 mg of active substance.

 The compounds of the invention can be administered according to the usual methods, for example enterally, orally, for example in the form of oral solutions, tablets or capsules, nasally, for example in the form of liquid sprays or powder for sprays and creams, or parenterally, for example in the form of solutions or injectable suspensions.

 For the compound of Example 2, an appropriate dose is between 3 and 10 mg administered 3 times daily, for example orally for the treatment of diabetes, ulcers and acromegaly.

 The compounds of the invention may be administered in any pharmaceutically acceptable form, for example in free form, i.e. free base, or when the compound is an acid, in the form of an acid. free, or in the form of a pharmaceutically acceptable salt. The salt may be an acid addition salt or, when the compound is an acid, a cationic salt. The compound can also be administered as a complex. The compounds may be in the form of a solvate, for example a hydrate.

 The present invention also relates to pharmaceutical compositions comprising a compound of the invention in a pharmaceutically acceptable form, in combination with at least one pharmaceutically acceptable carrier or diluent. Such compositions may be prepared according to the usual methods.

 One oral ampoule or solution for injection may contain per ml, for example 0.5 mg of the compound of Example 2 in the form of acetate, 11.45 mg of citric acid, 6.32 mg of NaOH, 4.5 mg NaCl.

 The subject of the present invention is also a compound of the invention suitable for any of the above-mentioned uses, including lowering the secretion of growth hormone, diabetes mellitus, reduction of gastric secretions and acromegaly for glucosylated derivatives of somatostatin type peptides.

 The present invention also relates to the use of a compound of the invention for the preparation of a medicament suitable for use in the treatment of the indications mentioned above, including the lowering of the secretion of the growth, diabetes mellitus, reduction of gastric secretions and acromegaly for glucosylated derivatives of somatostatin type peptides.

Claims (6)

 1. A process for preparing a peptide-alcohol which comprises at the C-terminus of the peptide chain 2 alcohol groups or an alcohol group and a thiol group, characterized in that the peptide-alcohol is prepared by acid hydrolysis of an acetal of the peptide-alcohol and a resin containing formylphenyl residues.  2. A process according to claim 1 for the preparation of a peptide-alcohol of formula

 in which Y represents the rest of a peptide-alcohol R1 means hydrogen or a methyl group and X is O or S, characterized in that a resin of formula Ir is hydrolyzed under acidic conditions

 which carries the peptide-alcohol, in which P represents the remainder of an insoluble synthetic resin, and Z represents a direct bond or a residue that connects the resin  with the acetylphenyl group acetalized, the acetalized CHO group is located in the meta or para position of the group Z and a number of acetalized formylphenyl groups are bonded to the molecule of the polymer resin.  3. A process according to claim 2, characterized in that

 is a polystyrene residue.  4. A process according to claim 2 or 3, characterized in that Z represents a group of the formula - (D) p-Ql-Q2- (E) q- in which Q1 represents the residue of a reactive group linked to the resin Q2 represents the rest of a reactive group linked to the formyl group  acetalized phenyl D represents a residue that connects the Q1 group to the polymer E represents a remainder that connects the Q2 group with the group  acetylated formylphenyl, and p and q independently of each other signify O or 1.  5. A method according to any one of claims 1 to 4, characterized in that Z represents a remainder

 in which R = H or CH3  m = 0 or 1 and P represents a polystyrene residue.  6. Compounds of formula Ir

 where is the rest of an insoluble synthetic resin  Z represents a direct bond or a remainder that connects  the resin with the acetylphenyl group acetalized  X means O or S  R1 is hydrogen or methyl, and  Y is the rest of a peptide-alcohol that can  contain protective groups, the acetalized CHO group is located in the meta or para position of the rest Z and a number of formylphenyl groups (acetalized) are attached to the polymer resin molecule, the compounds of formula Vr

 in which Pm represents the remainder of an insoluble synthetic resin, Z represents a direct bond or a residue that connects the resin  to the acetylphenyl acetal group, X means O or S, R1 means hydrogen or a methyl group and A represents a protecting group of the amino group and the group  acetal is in the meta or para position of the remainder Z, and the resin  comprises a number of acetalized formylphenyl groups, the compounds of formula VIr

 in which X is O or S, R1 is H or CH3, A represents a protecting group of the amino group, E represents a remainder that connects the Q'2 group with the group  acetyl formylphenyl, Q'2 represents a reactive group that can react with another  reactive group which is bound to a synthetic resin, and q is O or 1 where the acetal group is in the meta or para position of the group Q12- (E) q-> compounds of formula

 resin has a number of formylphenyl groups.  the formyl group is in the meta or para position of the Z group, and the  formylphenyl,  Z 'represents the rest that connects the resin with the group  wherein PD represents the remainder of an insoluble synthetic resin and