CA1183528A - Processes for the manufacture of novel glucose derivatives - Google Patents
Processes for the manufacture of novel glucose derivativesInfo
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- CA1183528A CA1183528A CA000403054A CA403054A CA1183528A CA 1183528 A CA1183528 A CA 1183528A CA 000403054 A CA000403054 A CA 000403054A CA 403054 A CA403054 A CA 403054A CA 1183528 A CA1183528 A CA 1183528A
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- methyl
- carbamoyl
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- ethyl
- propyl
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Abstract
Abstract The invention relates to processes for the manufac-ture of muramyl peptide derivatives of the formula I
(I) characterised in that at least one of the radicals R7, R9 and R13 represents lower alkyl, whilst the others represent hydrogen. The remaining substituents of the formula I are defined in claim 1. These compounds possess excellent immunomodulatory activity whilst, at the same time, especially when the substituent R7 represents lower alkyl, the pyrogenic side effect is greatly reduced as com-pared with the corresponding NH-compounds. They can there-fore be used in the form of pharmaceutical preparations.
In addition, the compounds according to the invention are used for the manufacture of the conjugates with antigens described in European Patent Application No. 79100513.5 and in Canadian Patent Application No. 321 992.
(I) characterised in that at least one of the radicals R7, R9 and R13 represents lower alkyl, whilst the others represent hydrogen. The remaining substituents of the formula I are defined in claim 1. These compounds possess excellent immunomodulatory activity whilst, at the same time, especially when the substituent R7 represents lower alkyl, the pyrogenic side effect is greatly reduced as com-pared with the corresponding NH-compounds. They can there-fore be used in the form of pharmaceutical preparations.
In addition, the compounds according to the invention are used for the manufacture of the conjugates with antigens described in European Patent Application No. 79100513.5 and in Canadian Patent Application No. 321 992.
Description
3~2~
4-11611/1-3/-~/s Canada Processes for the manufacture of novel glucose derivatives The invention relates to processes for the manu~
facture of N-alkylated muramyl peptides and derivatives thereof~ which can be used for the manufacture of pharma-ceutical preparations and in a method for the therapeutic treatment of the human or animal body.
The invention relates especially to a process for the manufacture of N-alkylated muramyl peptides of the formula I
4-11611/1-3/-~/s Canada Processes for the manufacture of novel glucose derivatives The invention relates to processes for the manu~
facture of N-alkylated muramyl peptides and derivatives thereof~ which can be used for the manufacture of pharma-ceutical preparations and in a method for the therapeutic treatment of the human or animal body.
The invention relates especially to a process for the manufacture of N-alkylated muramyl peptides of the formula I
2 R6 ~ ~-0 ~ 7 ~I) R3~CH (D) N-X-R2 \ 13 CoN-cH-coN-cH-cH2cH-R12 R7 (L) Rg (D) ~335~
in which X represents a carbonyl group, R represents hydrogen, alkyl, optionally substituted benzyl or acyl, R represents optionally substituted alkyl or carbocyclic aryl, R and R represent, independently of each other, hydrogen, alkyl, optionally substituted benzyl or acyl, R represents hydrogen or alkyl, at least one of the radicals R7, R and R represents lower alkyl, especially methyl, and the others represent hydrogen, R represents hydrogen, lower alkyl, free, esterified or etherified hydroxy-lower alkyl, free, esterified or etherified mercapto-lower alkyl, free or acylated amino-lower alkyl, cycloalkyl having 5 or 6 carbon atoms, cyclo-alkyl-lower alkyl in which the cycloalkyl radical contains 5 or 6 carbon atoms, optionally substituted aryl or aralkyl, nitrogen-containing heterocyclyl or hetero-cyclyl-lower alkyl, R and R together also represent alkylene having 3 or 4 carbon atoms, and the radicals 10~ Rll and R12 represent, independently of one another, an optionally esterified or amidated carboxy radical and Rll also represents hydrogen, salts of such compounds having salt-forming groups, pro-cesses for the manufacture of these compounds, pharmaceuti-cal agents containing such compounds and their use as phar-macologically active compounds.
The configuration of compounds in which R3, R
~L~83~2~
in which X represents a carbonyl group, R represents hydrogen, alkyl, optionally substituted benzyl or acyl, R represents optionally substituted alkyl or carbocyclic aryl, R and R represent, independently of each other, hydrogen, alkyl, optionally substituted benzyl or acyl, R represents hydrogen or alkyl, at least one of the radicals R7, R and R represents lower alkyl, especially methyl, and the others represent hydrogen, R represents hydrogen, lower alkyl, free, esterified or etherified hydroxy-lower alkyl, free, esterified or etherified mercapto-lower alkyl, free or acylated amino-lower alkyl, cycloalkyl having 5 or 6 carbon atoms, cyclo-alkyl-lower alkyl in which the cycloalkyl radical contains 5 or 6 carbon atoms, optionally substituted aryl or aralkyl, nitrogen-containing heterocyclyl or hetero-cyclyl-lower alkyl, R and R together also represent alkylene having 3 or 4 carbon atoms, and the radicals 10~ Rll and R12 represent, independently of one another, an optionally esterified or amidated carboxy radical and Rll also represents hydrogen, salts of such compounds having salt-forming groups, pro-cesses for the manufacture of these compounds, pharmaceuti-cal agents containing such compounds and their use as phar-macologically active compounds.
The configuration of compounds in which R3, R
~L~83~2~
3 ~
and/or R are different from hydrogen is indicated in formula I by ~D) and (L), respectively, at the respective centres of asymmetry.
The term "muramyl peptide" represents, in the strict sense, only compo~nds derived from muramic acid in which R is methyl. I~owever, unless expressly indicated to the contrary in the context, in this Application the term "muramyl peptides" ls applied also to compounds in which R represents hydrogen or an alkyl radical other than methyl and which, in the strict sense, should be termed normuramyl and ho~omuramyl peptide derivatives, respec-tively.
Alkyl is straight-chained or branched, having up to 18 carbon atoms and bonded in any position, but is especially lower alkyl.
As substituents of the optionally substituted alkyl group there come into consideration - especially free or functionally modified hydroxy or mercapto groups, such as etherified or esterified hydroxy or mercapto groups, for-example lower alkoxy or lower alkylmercapto groups, or halogen atoms or free or functionally modified carboxyl groups, such as lower alkoxycarbonyl or carbamoyl groups.
The substituted alkyl radical, such as the lower alkyl radical, can contain one, two or more identical or different substituents, especially free hydroxy groups or halogen atoms.
Carbocyclic aryl radicals are especially monocyclic, but also bicyclic, aryl radicals, especially phenyl, but also naphthyl. They may optionally be mono-, di- or poly-substituted, for example by lower alkyl groups, free, etherified or esterified hydroxy, for example lower alkoxy or lower alkylenedioxy, or halogen atoms, and/or trifluoro--methyl groups.
Aralkyl is especially aryl-lower alkyl, in which aryl has the meaning given above. Aryl-lower alkyl represents especially benzyl or phenethyl, in which the phenyl nucleus 3S;~
may be mono-, di- or poly-substituted.
Optionally substituted benzyl radicals are especially those benzyl radicals in which the aromatic nucleus is optionally mono-, di- or poly-substituted, for example by lower alkyl, free, etherified or esterified hydroxy or mercapto groups, for example lower alkoxy, lower alkylene-dioxy or lower alkylmercapto groups, or trifluoromethyl groups and/or halogen atoms.
Nitrogen-containing heterocyclyl is especially the radical of a 5- or 6-membered heterocyclic compound con-taining one or two nitrogen atoms in the ring. It can be unsaturated or saturated and, for example, contain a ~used-on phenyl radical. Examples that may be mentioned are the pyrrole, indane, pyridyl or imidazole ring.
An optionally esterified or amidated carboxyl group is especially the carboxyl group itself or a carboxyl group esterified by a lower alkanol, or a carbamoyl group which, at the nitrogen atom, is unsubstituted or mono- or di-substituted by alkyl, especially lower alkyl, aryl, espe-cially phenyl, or aralkyl, such as benzyl. The carbamoyl group can, however~ also carry an alkylene radical, such as the tetramethylene or pentamethylene radical. A carb-amoyl group R can also be substituted at the nitrogen atom by the carbamoy:Lmethyl group.
Acyl is especially an acyl radical of an organic acid, especially an organic carboxylic acid. ~hus, acyL is espe-cially alkanoyl, especially having from 2 to 18 carbon atoms, but more especially lower alkanoyl, or alternatively aroyl, such as naphthoyl-l, naphthoyl-2 and especially benzoyl, or benzoyl or naphthoyl substituted by halogen, lower alkyl, lower alkoxy, trifluoromethyl, hydroxy or lower alkanoyloxy, or alternatively an acyl radical of an organic sulphonic acid, for example an alkanesulphonic acid, espe-cially a lower alkanesulphonic acid, or an arylsulphonic acid, especially a phenylsulphonic acid, such as benzene-sulphonic acid or p-toluenesulphonic acid, optionally sub-35;~l~
stituted by lower alkyl or halogen, and also carbamoyl,for example unsubstituted carbamoyl, lower alkylcarbamoyl or arylcarbamoyl, such as rnethyl- or phenyl-carbamoyl.
Esterified or etherified hy~roxy is especially lower alkoxy or lower acyloxy, such as lower alkanoyloxy.
Esterified or etherifled mercapto is especially lower alkylmercapto or lower acylmercapto, such as lower alkanoyl-mercapto.
Acylated amino is especially lower alkanoylamino or carbamoylamino.
The radicals and compounds referred to as "lower" in the context of the present description and the patent claims contain preferably up to and including 7, and especially up to and including 4, carbon atoms.
Hereinbefore and hereinafter the general terms can have the following meanings:
Lower alkyl is, for example, n-propyl, n-butyl, iso-butyl, sec.-butyl or tert.-butyl, also n-pentyl, n-hexyl, isohexyl or n-heptyl and, especiaIly, methyl or ethyl.
In aryl-, cycloalkyl- or heterocyclyl-lower alkyl, the lower alkyl radical is especially methyl or ethyl, the aryl, cycloalkyl or heterocyclyl radical having the meanings given above.
Lower alkoxy is, for example, n-propoxy, n-butoxy, isobutoxy, sec.-butoxy or tert.-butoxy and, especially, methoxy or ethoxy.
Lower alkylmercapto is, for example, n-propyl-, n-butyl-, isobutyl-, sec.-butyl~ or tert.-butylmercapto and, especially, methylmercapto or ethylmercapto.
Lower alkylenedioxy is especially methylenedioxy, ethylenedioxy or propylenedioxy.
Halogen represents fluorine or bromine, but preferably represents chlorine.
Lower alkanoyl is especially propionyl or butyryl, but more especially acetyl.
The novel compounds of the present invention may be ~L835~
in the form oE mi~tures oE isomers or in the form of pure isomers.
Special mention should be made of compounds of the formula I in which R , R and R represent hydrogen, X represents carbonyl, R represents lower alkyl option-ally substituted by hydroxy or by methoxy, or phenyl option-ally substituted by hydroxy, methoxy, methyl, ethyl or by halogen, R and R represent hydrogen or methyl, R
and R represent lower alkyl or hydrogen, with the proviso that at least one of the radicals R , R and R represents lower allcyl, R represents methyl, ethyl, n-propyl, isopropyl, 2-methylpropyl, methylmercaptomethyl, hydroxymethyl, hydroxyethyl, phenyl, benæyl or p-hydroxy-benzyl~ Rlo' Rll and R represent carboxy, lower alkoxycarbonyl or carbamoyl and R also represents hydro~
gen. Of these compounds there are preferred those in which R represents lower alkyl, especially methyl, and R13 represents hydrogen.
The compounds preferred most of all are those mentioned in the Examples and homologues thereof.
The compounds of the formula I can be manufactured according to methods known per se. They can be obtained, for example, by condensing, in a manner known per se, a compound of the formula O ~ vV-R1 (II) ~-X-R2 R3--CH (D) R
\
COOH
~1 ~8~5'~3 2 3 13 eanings given above and R, RC' and ~ represent the radicals R , R and ~ , respectively, or represent a protecting group that can be split off readily, or a derivative thereof, with a compound of the formula 8 ~10 ~11 I - CH - COI - C~ICH2C~ - R12 R7 (L) R9 (D) (III) in which R an~ R have the meanings given above, R, RC' , R and R have the meanings of R , R , 8 and R , with the proviso that carboxy groups and, if desired, free hydroxyl groups present in these radicals are protected by protecting groups that can be split off readily, and splitting off optionally present protecting groups.
In this case, condensation is carried out, for example, by reacting the compound II in the form oE the activated carboxylic acid ~ith the amino compound III or by reacting the acid II with the compound III in which the amino group is in activated form. The activated carboxyl group can be, for example, an acid anhydride, preferably a mixed acid anhydride, such as an acid azide, an acid amide, such as an imidazolide or isoxazolide, or an activated ester. ~ctivated esters to be given special mention are cyanomethyl esters, carboxymethyl esters, p-nitrophenylthio esters, p-nitrophenyl esters, 2,4,5-trichlorophenyl esters, pentachlorophenyl esters, N-hydroxysuccinimide esters, N-hydroxyphthalimide esters, 8-hydroxy~uinoline esters, 2-hydroxy-1,2-dihydro-1-ethoxy-carbonylquinoline esters, N-hydroxypiperidine esters, or enol esters which are obtained with N-ethyl-5-phenyl-isoxa-zolium 3'-sulphonate. Activated esters can also be obtained, if desired, with a carbodiimide with the addition of N-hydroxysuccinimide or an unsubstituted or, for e~ample halogen-, methyl- or methoxy-substituted, l-hydroxy-benzo-triazole or 3-hydroxy-4-oxo~3,4-dihydro-benzo[d]1,2,3-tri-azine.
The amino group is activated, for example, by reaction with a phosphite amide.
Among the methods of reaction with activated esters there should be mentioned especially those with N-ethyl-5-phenyl-isoxazolium 3'-sulphonate (Woodward reagent K) or 2-ethoxy-1,2-dihydro-1-ethoxycarbonylquinoline or carbodi-imide.
Protecting groups that can be split off readily are those known from peptide and sugar chemistry. For carboxy groups there should be mentioned especially tert.-butyl, benzyl or benzhydryl, and for hydroxy groups especially acyl radicals, for example lower alkanoyl radicals, such as acetyl, aroyl radicals, such as benzoyl, and, more espe-cially, radicals derived from carbonic acid, such as benzyl-oxycarbonyl or lower alkoxycarbonyl, or alkyl, especially tert.-butyl, benzyl or tetrahydropyranyl each optionally substituted by nitro, lower alkoxy or halogen, or optionally substituted alkylidene radicals that link the oxygen atoms in the ~- and 6-position of the glucose moiety. Such alkylidene radicals are especially lower alkylidene radicals, especially ethylidene, isopropylidene or propyli-dene radicals, or alternatively benzylidene radicals that are optionally substituted, preferably in the p-position.
These protecting groups can be split off in a manner known per se. ~hey can be removed by hydrogenolysis, for example with hydrogen in the presence of a noble metal catalyst, such as a palladium or platinum catalyst, or by acid hydrolysis.
~35;~
_ 9 _ The starting materials used are known or can be manu-factured in a manner known per s Another method of manufacturing these novel starting materials comprises condensing, in a mannee known per se, a compound of the formula IV
,1--~ ~ rO-R1 (IV) R3-CH (D) R o I ' CON- CH - COOH
R7 (L) in which Rl, R2, R3, R, R, R and R have the meanings given above, with a compound of the formula R R
Rg (D) (V) in which R , R , R and R have the meanings 9 lO ll 12 given above, with the proviso that carboxyl groups present n the radicals R , R and R and, if desired, ~l 12 free hydroxy groups are protected by protecting groups that can be split off readily, and splitting off optionally pre-sent protecting groups.
The condensation is carried out in this case, for example, by reacting the compound IV in the form of the activated carboxylic acid with the amino compound V or by reacting the acid IV with the compound V in which the amino group is in activated form. The activated carboxyl group ~3S;~3 - 10 - .
can be, for example, an acid anhydride, preferably a mixed acid anhydride, an acid amide or an activated ester. As such, there come into consideration especially the acid anhydrides, amides or esters mentioned above. The amino group is activated, for example, by reaction with a phosphite amide.
The protecting groups that can be split of~ readily also correspond to those mentioned above. They can be split off in a manner known per se, for example by hydrogenolysis, for example with hydrogen in the presence of a noble metal catalyst, such as a palladium or platinum catalyst, or by aci~ hydrolysis.
The starting materials can be obtained in a manner known per se. For example, corresponding su~ars that are unsubstituted in the 3-position can be reacted with a halo-R -acetamido-R-acetic acid, or a compound of the formula II can be reacted, in the manner described above, with an amino-R -acetic acid in which the carboxyl group is protected, and the protecting group split off.
A further method of introducing the side chain located in the 3-position of the sugar moiety is to react a compound of the formula / ~
R~O~ ~ O-Rl (VI) ~--X--~
in which ~, R2, Rl, RC4, R6 and R13 have the meanings given above and hydroxy yroups optionally present therein are protected by a protecting group that can be split off readily, with a cornpound of the formula 33~;~8 (D) R8 1lO 1ll z fH CONCH - CON - C~ - CH2CH ~ R12 ~V~I) R3 R7 Rg in which Z represents a reactive esterified hydroxy group 3 7 8' 9' Rlo' Rll and R have the meanings given above, and to split off optionally pre-sent protecting groups.
A reactive esterified hydroxy group is especially a hydroxy group esterified by a strong inoeganic or organic acid, especially one esterified by a hydrohalic acid, such as hydrochloric, hydrobromic or hydriodic acid.
The protecting groups that can be split off readily correspond to those already mentioned above. They can be split off in a manner kno~n per se, for example by hydro-genolysis, for example with hydrogen in the presence of a noble metal catalyst, such as a palladium or platinum catalyst, or by acid hydrolysis.
Depending on the nature of their substituents, the present novel compounds are neutral, acidic or basic. If acid groups are present in excess, they form salts with ~ases, such as ammonium salts or salts with alkali metals or alkaline earth metals, for example sodium, potassium, calcium or magnesium. If, however, basic groups are pre-sent in excess, they form acid addition salts.
~ cid addition salts are especially pharmaceutically acceptable non-toxic acid addition salts, such as those with inor~anic acids, for example hydrochloric, hydrobromic, nitric, sulphuric or phosphoric acids, or with organic acids, such as organic carboxylic acids, for example acetic acid, propionic acid, glycolic acid, succinic acid, maleic ~ 83~
acid, hydroxymaleic acid, methylmaleic acid, fumaric acid, malic acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, salicylic acid, 4-arninosali-cylic acid, 2-phenoxybenzoic acid, 2-acetoxybenzoic acid, embonic acid, nicotinic acid or isonicotinic acid, or organic sulphonie acids, for example methanesulphonic acid, ethanesulphonic acid, 2 hydroxyethanesulphonic acid, ethane-1,2-disulphonic acid, benzenesulphonic acid, p-toluenesul-phonic acid or naphthalene-2-sulphonic acid, and also other acid addition salts which ean be used, or example, as intermediates, for example for purifying the free com~ounds or for the manufacture of other salts, and for characterisa-tion~ such as, for example, those with picric acid, picro-lonic acid, flavianic acid, phosphotungstic-acid, phospho-molybdic acid, chloroplatinic acid, Reinecke's acid or per-chlorie acid.
The muramyl peptides of the formula I according to the invention can be used both as intermediates and as end produets. For example, they are used for the manufaeture of the muramyl peptide~antigen conjugates deseribed in European Patent Application No. 79100513.5 (Publication Number 0003833).
Irhe muramyl peptides of the formula I as such are pharmacologically active and can be used as medicaments for the treatment of the human or animal body. In parti-eular, they have an immunomodulatory action, as can be demonstrated inter alia by the tests described below:
1. Potentia~ion of cellular immunity in vivo: intensifi-eation of delayed hyper-sensitivity to ovalbumin in guinea pigs _ _ -----Pirbright guinea pigs are immunised on day 0 withn mg of ovalbumin in complete Freund's adjuvant by injec-ting 0.1 ml of an antigen-adjuvant mixture into each hind paw. 4 weeks-later, skin reactions are triggered by intra-~ILi835~28 cutaneous injection of 100 ~g of ovalbumin in 0.1 mL of buffered physiological salt solution and are quantified on the basis of the reaction volume calculated 2~ hours thereafter by mean~ of the surface area of the erythema and the increase in skin thickness. The antigen-specific increase in the reaction volume observed after 2~1 hours (delayed reaction) is considered as a measure of cell-mediated immunity. Ovalburnin is too weak an immunogen to induce a delayed reaction on its own or in a water-oil emulsion with incomplete Freund's adjuvant (10 parts of ovalbumin solution in 0.9 % NaCl mixed with ~.5 parts of Bayol F and 1.5 part of Arlacel A) and, for effective immunisation, has to be administered in complete Freund's adjuvant to which mycobacteria have been added (5 mg of dead and lyophilised M butyricum per 10 ml of Bayol F/Arlacel A). To demonstrate the immunopotentiating action of the test substances, the latter can be added to the anti-gen-oil mixture, in doses of from 10 to 100 ~g, instead of the mycobacteria.
The glucosamine peptides according to the invention are capable of imitating the effect of the mycobacteria in the described test and of quantitatively surpassing that effect.
A significant potentiation of the delay2d reactivity to ovalbumin can also be achieved by administering the com-pounds of the described kind subcutaneously in sodium chloride solution, in doses of from 10 to 100 ~g per animal, for a few days after immunisation (for example, on day 0, 1, 2, 5, 6 and 7), rather than incorporating them into the antigen-oil mixture.
This shows that compounds of the described kind are capable of considerably increasing cellular immunity both in admixture with the antigen itself (adjuvant effect in the narrower sense) and when administered separately at a different time and at a different site from the antigen injection (systemic irnmunopotentiation).
3~i2~
2. Potentiation of humoral immunity ln vivo increase in the production of antibodies to bovine serumalbumin ~BS~) in mice . _ .
N~RI mice are immunised on day O by intraperitoneal (i.p.) injection of lO ~Lg of precipitate-free BSA. 9, 15 and 29 days later, serum samples are taken and examined for their content of anti-BSA-antibodies by means of a passive haemagglutination technique. In the dosage used, soluble BSA is sub-immunogenic for the recipient animals, i.e. it is unable to initiate anyl or is able to initiate only a very insignificant, production of antibodies. Addi-tional treatment of the mice with certain immunopotentiating substances before or after the administration of antigen results in an increase in the antibody titre in the serum.
The effect of the treatment is expressed by the score value achieved, that is to say, by the sum of log titre differences on the three days on which blood samples were taken. Compounds of the present invention are capable of significantly increasing the production of antibodies to BSA when administerecl intraperitoneally or subcutaneously (s.c.) at a dosage of 100 - 300 mg/kg/animal on five suc-cessive days (day O to 4) after immunisation with BSA.
~ he immunostimulating effect of the mentioned com-pounds is, in contrast to that of other bacterial immuno-leptics (for example, LPS from E. coli), antigen-dependent:
injection of the novel compounds results in an increase in the anti-BSA titre only in BSA-immunised and not, however, in non-immunised mice. Remarkably, s.c. admini-stration of the mentioned compounds is just as effective as i.p. administration, that is to say, the observed immuno-potentiating action is systemic and is not dependent on the stimulant being administered by the same route as the antigen or in admixture with the antigen, as is the case with conventional adjuvants.
The described tests show that compounds of the kind ~ ~35;~8 described are also capable of speciEically increasing humoral immunity, that they improve the immunological response to stimulus and that their immunopotentiating effects are due to a systemic activation of the immune mechanism.
3. Potentiation of humoral immunity in vitro: T-cell- -substituting effect in the antibody response of mice splenocyt~s to sheep erythrocytes (SE) To induce an antibody response, in many cases lymphocytes originating from the thymus (T-cells) are required. These cells co-operate with the precursors of antibody-forming lymphocytes (B-cells) and help them to react to stimulation by so-called T-dependent antigens with proliferation, differentiation and antibody synthesis.
Splenocyte suspensions of congenitally athymic nu/nu mice do not contain any fun-ctional T-cells and, for example, are not capable of forming any antl-SE-antibodies in vitro in the presence of SE. The compounds of the present invention, surprisingly, are capable of functionally replacing T-cells in such cultures and~making possible an antibody response to SE. The addition of these substances to nu/nu splenocyte cultures in the presence of SE resultsr within 4 days, in a considerable increase in the number of antibody-forming cells~ The findings show that the mentioned compounds are capable of increasing humoral anti-body formation ln vltro and of compensating a clefect of the T-cell system.
and/or R are different from hydrogen is indicated in formula I by ~D) and (L), respectively, at the respective centres of asymmetry.
The term "muramyl peptide" represents, in the strict sense, only compo~nds derived from muramic acid in which R is methyl. I~owever, unless expressly indicated to the contrary in the context, in this Application the term "muramyl peptides" ls applied also to compounds in which R represents hydrogen or an alkyl radical other than methyl and which, in the strict sense, should be termed normuramyl and ho~omuramyl peptide derivatives, respec-tively.
Alkyl is straight-chained or branched, having up to 18 carbon atoms and bonded in any position, but is especially lower alkyl.
As substituents of the optionally substituted alkyl group there come into consideration - especially free or functionally modified hydroxy or mercapto groups, such as etherified or esterified hydroxy or mercapto groups, for-example lower alkoxy or lower alkylmercapto groups, or halogen atoms or free or functionally modified carboxyl groups, such as lower alkoxycarbonyl or carbamoyl groups.
The substituted alkyl radical, such as the lower alkyl radical, can contain one, two or more identical or different substituents, especially free hydroxy groups or halogen atoms.
Carbocyclic aryl radicals are especially monocyclic, but also bicyclic, aryl radicals, especially phenyl, but also naphthyl. They may optionally be mono-, di- or poly-substituted, for example by lower alkyl groups, free, etherified or esterified hydroxy, for example lower alkoxy or lower alkylenedioxy, or halogen atoms, and/or trifluoro--methyl groups.
Aralkyl is especially aryl-lower alkyl, in which aryl has the meaning given above. Aryl-lower alkyl represents especially benzyl or phenethyl, in which the phenyl nucleus 3S;~
may be mono-, di- or poly-substituted.
Optionally substituted benzyl radicals are especially those benzyl radicals in which the aromatic nucleus is optionally mono-, di- or poly-substituted, for example by lower alkyl, free, etherified or esterified hydroxy or mercapto groups, for example lower alkoxy, lower alkylene-dioxy or lower alkylmercapto groups, or trifluoromethyl groups and/or halogen atoms.
Nitrogen-containing heterocyclyl is especially the radical of a 5- or 6-membered heterocyclic compound con-taining one or two nitrogen atoms in the ring. It can be unsaturated or saturated and, for example, contain a ~used-on phenyl radical. Examples that may be mentioned are the pyrrole, indane, pyridyl or imidazole ring.
An optionally esterified or amidated carboxyl group is especially the carboxyl group itself or a carboxyl group esterified by a lower alkanol, or a carbamoyl group which, at the nitrogen atom, is unsubstituted or mono- or di-substituted by alkyl, especially lower alkyl, aryl, espe-cially phenyl, or aralkyl, such as benzyl. The carbamoyl group can, however~ also carry an alkylene radical, such as the tetramethylene or pentamethylene radical. A carb-amoyl group R can also be substituted at the nitrogen atom by the carbamoy:Lmethyl group.
Acyl is especially an acyl radical of an organic acid, especially an organic carboxylic acid. ~hus, acyL is espe-cially alkanoyl, especially having from 2 to 18 carbon atoms, but more especially lower alkanoyl, or alternatively aroyl, such as naphthoyl-l, naphthoyl-2 and especially benzoyl, or benzoyl or naphthoyl substituted by halogen, lower alkyl, lower alkoxy, trifluoromethyl, hydroxy or lower alkanoyloxy, or alternatively an acyl radical of an organic sulphonic acid, for example an alkanesulphonic acid, espe-cially a lower alkanesulphonic acid, or an arylsulphonic acid, especially a phenylsulphonic acid, such as benzene-sulphonic acid or p-toluenesulphonic acid, optionally sub-35;~l~
stituted by lower alkyl or halogen, and also carbamoyl,for example unsubstituted carbamoyl, lower alkylcarbamoyl or arylcarbamoyl, such as rnethyl- or phenyl-carbamoyl.
Esterified or etherified hy~roxy is especially lower alkoxy or lower acyloxy, such as lower alkanoyloxy.
Esterified or etherifled mercapto is especially lower alkylmercapto or lower acylmercapto, such as lower alkanoyl-mercapto.
Acylated amino is especially lower alkanoylamino or carbamoylamino.
The radicals and compounds referred to as "lower" in the context of the present description and the patent claims contain preferably up to and including 7, and especially up to and including 4, carbon atoms.
Hereinbefore and hereinafter the general terms can have the following meanings:
Lower alkyl is, for example, n-propyl, n-butyl, iso-butyl, sec.-butyl or tert.-butyl, also n-pentyl, n-hexyl, isohexyl or n-heptyl and, especiaIly, methyl or ethyl.
In aryl-, cycloalkyl- or heterocyclyl-lower alkyl, the lower alkyl radical is especially methyl or ethyl, the aryl, cycloalkyl or heterocyclyl radical having the meanings given above.
Lower alkoxy is, for example, n-propoxy, n-butoxy, isobutoxy, sec.-butoxy or tert.-butoxy and, especially, methoxy or ethoxy.
Lower alkylmercapto is, for example, n-propyl-, n-butyl-, isobutyl-, sec.-butyl~ or tert.-butylmercapto and, especially, methylmercapto or ethylmercapto.
Lower alkylenedioxy is especially methylenedioxy, ethylenedioxy or propylenedioxy.
Halogen represents fluorine or bromine, but preferably represents chlorine.
Lower alkanoyl is especially propionyl or butyryl, but more especially acetyl.
The novel compounds of the present invention may be ~L835~
in the form oE mi~tures oE isomers or in the form of pure isomers.
Special mention should be made of compounds of the formula I in which R , R and R represent hydrogen, X represents carbonyl, R represents lower alkyl option-ally substituted by hydroxy or by methoxy, or phenyl option-ally substituted by hydroxy, methoxy, methyl, ethyl or by halogen, R and R represent hydrogen or methyl, R
and R represent lower alkyl or hydrogen, with the proviso that at least one of the radicals R , R and R represents lower allcyl, R represents methyl, ethyl, n-propyl, isopropyl, 2-methylpropyl, methylmercaptomethyl, hydroxymethyl, hydroxyethyl, phenyl, benæyl or p-hydroxy-benzyl~ Rlo' Rll and R represent carboxy, lower alkoxycarbonyl or carbamoyl and R also represents hydro~
gen. Of these compounds there are preferred those in which R represents lower alkyl, especially methyl, and R13 represents hydrogen.
The compounds preferred most of all are those mentioned in the Examples and homologues thereof.
The compounds of the formula I can be manufactured according to methods known per se. They can be obtained, for example, by condensing, in a manner known per se, a compound of the formula O ~ vV-R1 (II) ~-X-R2 R3--CH (D) R
\
COOH
~1 ~8~5'~3 2 3 13 eanings given above and R, RC' and ~ represent the radicals R , R and ~ , respectively, or represent a protecting group that can be split off readily, or a derivative thereof, with a compound of the formula 8 ~10 ~11 I - CH - COI - C~ICH2C~ - R12 R7 (L) R9 (D) (III) in which R an~ R have the meanings given above, R, RC' , R and R have the meanings of R , R , 8 and R , with the proviso that carboxy groups and, if desired, free hydroxyl groups present in these radicals are protected by protecting groups that can be split off readily, and splitting off optionally present protecting groups.
In this case, condensation is carried out, for example, by reacting the compound II in the form oE the activated carboxylic acid ~ith the amino compound III or by reacting the acid II with the compound III in which the amino group is in activated form. The activated carboxyl group can be, for example, an acid anhydride, preferably a mixed acid anhydride, such as an acid azide, an acid amide, such as an imidazolide or isoxazolide, or an activated ester. ~ctivated esters to be given special mention are cyanomethyl esters, carboxymethyl esters, p-nitrophenylthio esters, p-nitrophenyl esters, 2,4,5-trichlorophenyl esters, pentachlorophenyl esters, N-hydroxysuccinimide esters, N-hydroxyphthalimide esters, 8-hydroxy~uinoline esters, 2-hydroxy-1,2-dihydro-1-ethoxy-carbonylquinoline esters, N-hydroxypiperidine esters, or enol esters which are obtained with N-ethyl-5-phenyl-isoxa-zolium 3'-sulphonate. Activated esters can also be obtained, if desired, with a carbodiimide with the addition of N-hydroxysuccinimide or an unsubstituted or, for e~ample halogen-, methyl- or methoxy-substituted, l-hydroxy-benzo-triazole or 3-hydroxy-4-oxo~3,4-dihydro-benzo[d]1,2,3-tri-azine.
The amino group is activated, for example, by reaction with a phosphite amide.
Among the methods of reaction with activated esters there should be mentioned especially those with N-ethyl-5-phenyl-isoxazolium 3'-sulphonate (Woodward reagent K) or 2-ethoxy-1,2-dihydro-1-ethoxycarbonylquinoline or carbodi-imide.
Protecting groups that can be split off readily are those known from peptide and sugar chemistry. For carboxy groups there should be mentioned especially tert.-butyl, benzyl or benzhydryl, and for hydroxy groups especially acyl radicals, for example lower alkanoyl radicals, such as acetyl, aroyl radicals, such as benzoyl, and, more espe-cially, radicals derived from carbonic acid, such as benzyl-oxycarbonyl or lower alkoxycarbonyl, or alkyl, especially tert.-butyl, benzyl or tetrahydropyranyl each optionally substituted by nitro, lower alkoxy or halogen, or optionally substituted alkylidene radicals that link the oxygen atoms in the ~- and 6-position of the glucose moiety. Such alkylidene radicals are especially lower alkylidene radicals, especially ethylidene, isopropylidene or propyli-dene radicals, or alternatively benzylidene radicals that are optionally substituted, preferably in the p-position.
These protecting groups can be split off in a manner known per se. ~hey can be removed by hydrogenolysis, for example with hydrogen in the presence of a noble metal catalyst, such as a palladium or platinum catalyst, or by acid hydrolysis.
~35;~
_ 9 _ The starting materials used are known or can be manu-factured in a manner known per s Another method of manufacturing these novel starting materials comprises condensing, in a mannee known per se, a compound of the formula IV
,1--~ ~ rO-R1 (IV) R3-CH (D) R o I ' CON- CH - COOH
R7 (L) in which Rl, R2, R3, R, R, R and R have the meanings given above, with a compound of the formula R R
Rg (D) (V) in which R , R , R and R have the meanings 9 lO ll 12 given above, with the proviso that carboxyl groups present n the radicals R , R and R and, if desired, ~l 12 free hydroxy groups are protected by protecting groups that can be split off readily, and splitting off optionally pre-sent protecting groups.
The condensation is carried out in this case, for example, by reacting the compound IV in the form of the activated carboxylic acid with the amino compound V or by reacting the acid IV with the compound V in which the amino group is in activated form. The activated carboxyl group ~3S;~3 - 10 - .
can be, for example, an acid anhydride, preferably a mixed acid anhydride, an acid amide or an activated ester. As such, there come into consideration especially the acid anhydrides, amides or esters mentioned above. The amino group is activated, for example, by reaction with a phosphite amide.
The protecting groups that can be split of~ readily also correspond to those mentioned above. They can be split off in a manner known per se, for example by hydrogenolysis, for example with hydrogen in the presence of a noble metal catalyst, such as a palladium or platinum catalyst, or by aci~ hydrolysis.
The starting materials can be obtained in a manner known per se. For example, corresponding su~ars that are unsubstituted in the 3-position can be reacted with a halo-R -acetamido-R-acetic acid, or a compound of the formula II can be reacted, in the manner described above, with an amino-R -acetic acid in which the carboxyl group is protected, and the protecting group split off.
A further method of introducing the side chain located in the 3-position of the sugar moiety is to react a compound of the formula / ~
R~O~ ~ O-Rl (VI) ~--X--~
in which ~, R2, Rl, RC4, R6 and R13 have the meanings given above and hydroxy yroups optionally present therein are protected by a protecting group that can be split off readily, with a cornpound of the formula 33~;~8 (D) R8 1lO 1ll z fH CONCH - CON - C~ - CH2CH ~ R12 ~V~I) R3 R7 Rg in which Z represents a reactive esterified hydroxy group 3 7 8' 9' Rlo' Rll and R have the meanings given above, and to split off optionally pre-sent protecting groups.
A reactive esterified hydroxy group is especially a hydroxy group esterified by a strong inoeganic or organic acid, especially one esterified by a hydrohalic acid, such as hydrochloric, hydrobromic or hydriodic acid.
The protecting groups that can be split off readily correspond to those already mentioned above. They can be split off in a manner kno~n per se, for example by hydro-genolysis, for example with hydrogen in the presence of a noble metal catalyst, such as a palladium or platinum catalyst, or by acid hydrolysis.
Depending on the nature of their substituents, the present novel compounds are neutral, acidic or basic. If acid groups are present in excess, they form salts with ~ases, such as ammonium salts or salts with alkali metals or alkaline earth metals, for example sodium, potassium, calcium or magnesium. If, however, basic groups are pre-sent in excess, they form acid addition salts.
~ cid addition salts are especially pharmaceutically acceptable non-toxic acid addition salts, such as those with inor~anic acids, for example hydrochloric, hydrobromic, nitric, sulphuric or phosphoric acids, or with organic acids, such as organic carboxylic acids, for example acetic acid, propionic acid, glycolic acid, succinic acid, maleic ~ 83~
acid, hydroxymaleic acid, methylmaleic acid, fumaric acid, malic acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, salicylic acid, 4-arninosali-cylic acid, 2-phenoxybenzoic acid, 2-acetoxybenzoic acid, embonic acid, nicotinic acid or isonicotinic acid, or organic sulphonie acids, for example methanesulphonic acid, ethanesulphonic acid, 2 hydroxyethanesulphonic acid, ethane-1,2-disulphonic acid, benzenesulphonic acid, p-toluenesul-phonic acid or naphthalene-2-sulphonic acid, and also other acid addition salts which ean be used, or example, as intermediates, for example for purifying the free com~ounds or for the manufacture of other salts, and for characterisa-tion~ such as, for example, those with picric acid, picro-lonic acid, flavianic acid, phosphotungstic-acid, phospho-molybdic acid, chloroplatinic acid, Reinecke's acid or per-chlorie acid.
The muramyl peptides of the formula I according to the invention can be used both as intermediates and as end produets. For example, they are used for the manufaeture of the muramyl peptide~antigen conjugates deseribed in European Patent Application No. 79100513.5 (Publication Number 0003833).
Irhe muramyl peptides of the formula I as such are pharmacologically active and can be used as medicaments for the treatment of the human or animal body. In parti-eular, they have an immunomodulatory action, as can be demonstrated inter alia by the tests described below:
1. Potentia~ion of cellular immunity in vivo: intensifi-eation of delayed hyper-sensitivity to ovalbumin in guinea pigs _ _ -----Pirbright guinea pigs are immunised on day 0 withn mg of ovalbumin in complete Freund's adjuvant by injec-ting 0.1 ml of an antigen-adjuvant mixture into each hind paw. 4 weeks-later, skin reactions are triggered by intra-~ILi835~28 cutaneous injection of 100 ~g of ovalbumin in 0.1 mL of buffered physiological salt solution and are quantified on the basis of the reaction volume calculated 2~ hours thereafter by mean~ of the surface area of the erythema and the increase in skin thickness. The antigen-specific increase in the reaction volume observed after 2~1 hours (delayed reaction) is considered as a measure of cell-mediated immunity. Ovalburnin is too weak an immunogen to induce a delayed reaction on its own or in a water-oil emulsion with incomplete Freund's adjuvant (10 parts of ovalbumin solution in 0.9 % NaCl mixed with ~.5 parts of Bayol F and 1.5 part of Arlacel A) and, for effective immunisation, has to be administered in complete Freund's adjuvant to which mycobacteria have been added (5 mg of dead and lyophilised M butyricum per 10 ml of Bayol F/Arlacel A). To demonstrate the immunopotentiating action of the test substances, the latter can be added to the anti-gen-oil mixture, in doses of from 10 to 100 ~g, instead of the mycobacteria.
The glucosamine peptides according to the invention are capable of imitating the effect of the mycobacteria in the described test and of quantitatively surpassing that effect.
A significant potentiation of the delay2d reactivity to ovalbumin can also be achieved by administering the com-pounds of the described kind subcutaneously in sodium chloride solution, in doses of from 10 to 100 ~g per animal, for a few days after immunisation (for example, on day 0, 1, 2, 5, 6 and 7), rather than incorporating them into the antigen-oil mixture.
This shows that compounds of the described kind are capable of considerably increasing cellular immunity both in admixture with the antigen itself (adjuvant effect in the narrower sense) and when administered separately at a different time and at a different site from the antigen injection (systemic irnmunopotentiation).
3~i2~
2. Potentiation of humoral immunity ln vivo increase in the production of antibodies to bovine serumalbumin ~BS~) in mice . _ .
N~RI mice are immunised on day O by intraperitoneal (i.p.) injection of lO ~Lg of precipitate-free BSA. 9, 15 and 29 days later, serum samples are taken and examined for their content of anti-BSA-antibodies by means of a passive haemagglutination technique. In the dosage used, soluble BSA is sub-immunogenic for the recipient animals, i.e. it is unable to initiate anyl or is able to initiate only a very insignificant, production of antibodies. Addi-tional treatment of the mice with certain immunopotentiating substances before or after the administration of antigen results in an increase in the antibody titre in the serum.
The effect of the treatment is expressed by the score value achieved, that is to say, by the sum of log titre differences on the three days on which blood samples were taken. Compounds of the present invention are capable of significantly increasing the production of antibodies to BSA when administerecl intraperitoneally or subcutaneously (s.c.) at a dosage of 100 - 300 mg/kg/animal on five suc-cessive days (day O to 4) after immunisation with BSA.
~ he immunostimulating effect of the mentioned com-pounds is, in contrast to that of other bacterial immuno-leptics (for example, LPS from E. coli), antigen-dependent:
injection of the novel compounds results in an increase in the anti-BSA titre only in BSA-immunised and not, however, in non-immunised mice. Remarkably, s.c. admini-stration of the mentioned compounds is just as effective as i.p. administration, that is to say, the observed immuno-potentiating action is systemic and is not dependent on the stimulant being administered by the same route as the antigen or in admixture with the antigen, as is the case with conventional adjuvants.
The described tests show that compounds of the kind ~ ~35;~8 described are also capable of speciEically increasing humoral immunity, that they improve the immunological response to stimulus and that their immunopotentiating effects are due to a systemic activation of the immune mechanism.
3. Potentiation of humoral immunity in vitro: T-cell- -substituting effect in the antibody response of mice splenocyt~s to sheep erythrocytes (SE) To induce an antibody response, in many cases lymphocytes originating from the thymus (T-cells) are required. These cells co-operate with the precursors of antibody-forming lymphocytes (B-cells) and help them to react to stimulation by so-called T-dependent antigens with proliferation, differentiation and antibody synthesis.
Splenocyte suspensions of congenitally athymic nu/nu mice do not contain any fun-ctional T-cells and, for example, are not capable of forming any antl-SE-antibodies in vitro in the presence of SE. The compounds of the present invention, surprisingly, are capable of functionally replacing T-cells in such cultures and~making possible an antibody response to SE. The addition of these substances to nu/nu splenocyte cultures in the presence of SE resultsr within 4 days, in a considerable increase in the number of antibody-forming cells~ The findings show that the mentioned compounds are capable of increasing humoral anti-body formation ln vltro and of compensating a clefect of the T-cell system.
4. Selective mitogenicity for B-cells: proliferation-promotinq effect in B-lymphocyte cultures SUSpQnSions of highly enriched B-lymphocytes (lymph node cells of congenitally athymic nu/nu mice~ and sub-stantially pure, immature and mature T-lymphocytes (thymus 3S~
- lG -cells and cortisone-resistant thymus cells, i.e. those per-sisting 48 hours after a cortisone injection, of Balb/c mice) are incubated in the presence of the test substances for 3 days. The incorporation of M -thymidine into the lymphocytes during the last 18 hours of the culture period is considered as a measure of the prolifeeation activity.
The compounds according to the invention are mitogenic for B-lymphocytes (i.e. for the precursors of the antibody-forming cells) but not for 'r-lymphocytes.
They are thus capable of stimulating the proliferation of lymphocytes involved in the humoral immune response.
- lG -cells and cortisone-resistant thymus cells, i.e. those per-sisting 48 hours after a cortisone injection, of Balb/c mice) are incubated in the presence of the test substances for 3 days. The incorporation of M -thymidine into the lymphocytes during the last 18 hours of the culture period is considered as a measure of the prolifeeation activity.
The compounds according to the invention are mitogenic for B-lymphocytes (i.e. for the precursors of the antibody-forming cells) but not for 'r-lymphocytes.
They are thus capable of stimulating the proliferation of lymphocytes involved in the humoral immune response.
5. Tolerability By comparison with the muramyl peptides of the formula I belonging to the state of the art in which R , R
and R represent hydrogen, the compounds of the formula I according to the invention, especially those in which the radical R represents lower alkyl, especially methyl, and more especially those in which R represents lower alkyl and R represents methyl, are distinguished by a greatly reduced pyrogenic action or, rather, one which is undetectable or tolerable in the dosages that come into consideration for practical applications, while the immuno-modulating properties remain roughly the same or are altered, usually increased, to a minor extent. These findings are of great practical significance because the pyrogenic action of the hydrophilic muramyl peptides known before the present invention in mammalsother than and as opposed ~ roden-ts is considerable and their therapeutic range is therefore too narrow to allow them to be used safe]y without const~nt medical supervision. In particular, the pyrogenic side effect could, in unfavourable circumstances, lead to thermal shock and, for that reason, certain forms of admini-stration, such as intravenous administration, are out oE
the question.
sz~
The muramyl peptides according to the invention are not toxic even when 300 mg/kg are administered i.p. to mice 5 times.
Testing for pyrogenicity was carried out on rabbits in the manner prescribed in European Pharmacopoeia, vol. 2, 1971, pages 56-59. ~ccording to this, for example even when a h gh dose, such as 10 mg/kg, of N-acetyl-muramyl-L-(~-methyl)-~-aminobutyryl-D-isoglutamine is a~ministered, no pyrogenic effect can be observed.
In the light of the state of the art, the high adju-vant activity of the compounds according to the invention in comparison with their greatly reduced pyrogenic action is surprising since, according to previous investigations, high adjuvant activity W3S always associated with a high pyrogenicity and vice versa (cf. Sh~ Kotani et al, Biken Journal 19, 9-13[1976]).
The compounds according to the invention are capable, on the one hand in admixture with an antigen, of increasing the immunogenicity of the latter and, on the other hand when administered systemically, of intensifying the immuno-logical reactivity of the treated organism. In so doing, the mentioned substances are able to promote both the cellular and the humoral immunity and to activate the lymphocytes responsible for the formation of antibodies.
The novel compounds can therefore be used as adjuvants in admixture with vaccines to improve the success of vac-cination and to improve the protection against infection mediated by humoral antibodies and/or cellular immunity against bacterial, viral or parasitic causative organisms.
Finally, the described compounds in admixture with a wide variety of antigens are suitable as adjuvants in the experimental and industrial manufacture of antisera for therapy and diagnostics and in the induction of immuno-logically activated lymphocyte populations for cell trans-fer processes.
Furthermore, the novel compounds can be used also s~
without the simultaneous administration of antigens to pro-mote immune reactions already proceeding subliminally in humans and animals. The compounds are accordingly suitable especially for the stimulation of the body's own defences, for example in chronic and acute infections or in the case of selective (antigen-speciEic) immunological defects, and in hereditary and also in acquired general (i.e. not antigen-specific) immunologically defective conditions, such as occur in old age, in the course of serious primary illnesses and especiaLly arter therapy with ionising radia-tion or with hormones having an immunosuppressive action.
The mentioned substances can therefore be administered, preferably also in combination with antibiotics admini-stered to combat infections, with chemotherapeutic agents and other pharmaceutical remedies, to counteract immunolo-gical damage. Finally, the described substances are suit-able also for the general prophylaxis of infectious diseases in humans and animals.
The present invention relates also to pharmaceutical preparations that contain compounds of the formula I. The pharmaceutical preparations according to the invention are those suitable for enteral, such as oral or rectal, admini-stration, and for parenteral administration to warm-blooded animals, which contain the pharmacological active sub-stance on its own or together with a pharmaceutically accep~
table carrier. The dosage of the active substance depends on the species of warm-blooded animal, the age and individual condition and also on the method of administra-tion and the nature of the illness treated or the nature of the desired effect.
Accordingly, the daily doses to warm-blooded animals of 70 kg body weight lie, in the case of parenteral administration, between 0.5 and 100 mg, preferably between l and 50 mg, for example 5 mg, and, in the case of oral administration, between 1 and 1500 mg, preferabLy between 5 and 750 mg, for example 50 mg~
~352~3 An appropriate fraction of the amount mentioned ~bove is administered one or more times daily, preEerably from one to three times.
The novel pharmaceutical preparations contain from approximately 10 % to approximately 95 ~, preferably from approximately 20 % to approximately 90 ~, of the active substance. Pharmaceutical preparations according to the invention can, for example, be in unit dose form, such as deagées, tablets, capsules, suppositories or ampoules.
The pharmaceutical preparations of the present invention are prepared in a manner known per se, Eor example by means of conventional mixing, granulating, coating, dis-solving or lyophilising processes~ Pharmaceutical prepara-tions for oral use can be obtained by combining the active substance with solid carriers, if desired granulating a resulting mixture and processing the mixture or granulate, if desired or necessary after the addition of suitable adjuncts, to form tablets or dragée cores.
Suitable carriers are especially fillers, such as sugars, for example :Lactose, saccharose, mannitol or sorbitol, cellulose preparations and/or calcium phosphates, for example tricalcium phosphate or calcium hydrogen phosphate, also binders, such as starch pastes, using, for example, corn, wheat, rice or potato starch, gelatin, tragacanth, methylcellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose and/or polyvinylpyrrolidone, and/or, if desired, disintegrators, such as the above-mentioned starches, also carboxymethyl starch, crosslinked polyvinylpyrrolidone, agar, alginic acid or a salt thereof, such as sodium alginate. Adjuncts are especially flow-regulators and lubricants, for example silica, talc, stearic acid or salts thereof, such as magnesium or calcium stearate, and/or polyethylene glycol. Dragée cores are provided with suitable coatings that may be resistant to gastric juice, there being used, inter alia, concentrated sugar solutions that optionally contain gum arabic, talc, polyvinylpyrrolidone, polyethylene glycol and/or titanium dioxide, lacquer solutions in suitable organic solvents or solvent mixtures or, for the manufacture of coatings that are resistant to gastric juice, solutions of suitable cellulose preparations, such as acetylcellulose phthalate or hydroxypropylmethylcellulose phthalate. Colouring sub-stances or pigments may be added to the tablets or dragée coatings, for example for identification or for indicating different doses of active substance.
In certain cases, liposomal forms of administration are especially suitable.
The following Examples illustrate the invention without limiting it in any form. The R values were ascertained on silica gel thin-layer plates manufactured by Merck. Temperatures are given in degrees Centigrade.
335;~
Example 1 7.12 g (20 mmol) of the sodium salt of benzyl-2-acetylamino-3-0-carboxymethyl-2-deoxy-4,6-0-isopropyli-dene-a-D-glucopyranoSide are suspended in 60 ml oE
dimethylformamide. After the addition of 6.46 g (20 mmol) of L-(N-methyl)-alanyl-D~isoglutamine-~-tert.-butyl ~ster hydrochloride, 5.91 g (24 mmol) of ~EDQ (2-ethoxy-N-ethox~carbonyl-1,2-dihydro-quinoline) are added to the mixture and the whole is stirred for 24 hours at room temperature. The suspension is concentrated by evaporation at 30 in vacuo, the residue is taken up in 100 ml of ethyl acetate and extracted successively, at low temperature, with 2~ potassium bicarbonate solution and 2N citric acid solution, i5 finally washed neutral with water and, after drying over sodium sulphate, concentrated by evaporation. ~he by-products still present are remoYed by chromatography over silica gel (toluene/acetone = 7/3).
Having been concentrated by evaporation, the eluate yields 7.7 9 (57 %) of henzyl-2-acetylamino-~,6-O~isopropylidene-3-O-{~L-1-(D-l~carbamoyl-3-tert.-butoxycarbonyl-propyl)-carbamoyl-ethyl]-N-methyl-carbamoylmethyl}-2-deoxy-a-D-glucopyranoside.
[~]D = +66 (c = 1.1; methanol), R = 0.81 (ethyl acetate/n-butanol/pyridine/acetic acid/
water = 42/21/21/6/lO), R = Or 56 (n~butanol/acetic acid/water = 75/7.5/21), R = ~.30 tchloroform/isopropanol/acetic acid = 70/8/2)o C H N O calc. C 58.39 1~ 7.42 ~ 8.25 33 50 ~ 11 (678.80) found C 58.01 H 7.50 N 8.20 The protecting grou?s are split off from the resulting product as follows:
7.60 g (11.1 mmol) of benzyl-2-acetylamino-4,6-0-iso-propylidene-3-0-{[L-l-(D-l-carbamoyl-3-tert.-butoxycarbonyl-propyl)-carbamoyl-ethyl]-N-methyl-carbamoylmethyl}-2-deoxy-~-D-glucopyranoside are dissolved in 80 ml of cold 95 ~ trifluoroacetic acid, the solu~ion is stirred for 15 minutes at ~ C and then 1.1 litre of ether/petroleum ether = 9/1 is added. The oily precipitate is triturated several times with fresh mixture until a solid deposit forms. The latter is dissolved in 30 ml of water and filtered over 20 ml of weakly basic ion-exchanger (Dowex-3) in acetate form to remove the trifluoroacetic acid. The column is subsequently rinsed with water and the combined eluates are lyophilised. 4.95 9 (85 %) of benzyl-2-acetylamino-3-0-{[L-l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-ethyl]-N-methyl-carbamoylmethyl}-2--deoxy-~-D-glucopyranoside are obtained.
[a]20 = +~5 (c = 1-2; H20), R - 0.28 (n-butanol/acetic acid/water - 75/7.5/21), R = 0.10 (chloroform/methanol/water=70/30/5).
4.65 g ~80 mmol) of the resulting benzyl-2-acetyl-amino-3-0-{[L-l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-ethyl]-~-methyl-carbamoylmethyl}-2-deoxy-a-D-gluco~yranoside are dissolved in 100 ml of methanol, 4.6 9 of palladium-on-carbon (10 %) are added and the mixture is treated exhaus-tively with hydrogen Eor 2 days. The catalyst is Eiltered off and the filtrate is concentrated by evaporation at room temperature. The residue is dissolved in 50 ml of water, extracted several times with n-butanol and, after evapora-ting the butanol, the aqusous phase is lyophilised. 3.2 9 (81.4 %) oE 2-acetylamino-3-0-{[L-l-(~-l-carbamoyl-3-carboxy-propyl)-carbamoyl-ethyl~-N-methyl-carbamoylmethyl}-2~deoxy-D-glucose are obtained.
[a]DO = -19 (c = 1.1; methanol), f = 0.20 (ethyl acetate/n-butanol/pyridine/acetic acid/
water = 42/21/21/6/10), ~ = 0.10 (n-butanol/acetic acid/water = 75/7.5/21), Rf = 0.45 (acetonitrile/water = 3/1) C H N 0 .1.2H 0 calc. C ~ 36 ~ 6.78 N 10.~9 19 32 ~ 11 2 (514.46) found C 44.32 H 6.75 ~ 10.79 The coupling components (muramic acid derivative and dipeptide) used as starting materials are obtained as follows:
1.2 g of sodium hydride pract~ (Fluka) is added to a solution of 8.0 g of benzyl-2-acetylamino-2-deoxy-4,6-0-iSopropylidene-a-D-glucopyranoside in 80 ml of acetonitrile and the mixture is stirred at 40 C for 2 hours. The reac-o otion mixture is subsequently cooled to from -5 to -10 and 5.2 g of bromoacetic acid methyl ester are added. The mixture is stirred for a further 30 minutes at -5 C, 10 ml of methanol are added and, after a further 10 minutes, the mixture is neutralised with acetic acid. The reaction mix-ture is concentrated to dryness by evaporation and the resi-due is partitioned between distilled water and ether. The ether solution is washed with distilled water, dried over magnesium sulphate and concentrated to dryness by evapora-tion. The resulting residue is crystallised from ether/
petroleum ether. Benzyl-2-acetylamino-3-0-methoxycarbonyl-2-deoxy-4,6-0-isopropylidene-a-D-glucopyranoside is obtained.
[~]20 = +150 ~ ~ (c = 0.875, C~C13), m.p. 122-3 C.
15 ml of lN sodium hydroxide solution are added to a solution of 4.2 g of benzyl-2-acetylamino-3-0-methoxy-carbonylmethyl-2-deoxy-4,6-0-isopropylidene-a-D~gluco-pyranoside in 20 ml of methanol and the resulting solution is left to stand at room temperature for one hour. Then, 5 ml of lN hydrochloric acid are added dropwise, while stirring, and the solution is concentrated to dryness by evaporation. The resulting po~der is dried at a high tem-perature under a high vacuum and constitutes 10 mmol of the 33~
- 2~ -~sodium salt of ben~yl-2-acetylamino-3-0-carboxy,nethyl-2-deoxy-4,6-0-isopropylidene~~-D-glucopyranoside.
18.5 g t43.9 mmol) of N-benzyloxycarbonyl-N-rllethyl-L-alanyl-D-isoglutamine-y-tert.-butyl ester are dissolved in 150 ml of methanol, 4 g of palladium-on-carbon tlO ~) are added and the mixture is hydrogenated at pE~ 4.5 tpH-stat) in the presence of 0.65N methanolic hydrochloric acid.
The catalyst is filtered off, the filtrate is concentrated by evaporation in vacuo and the residue is crystallised from 25 ml of dimethox~ethane and 100 ml of ether. 11.6 g (82 ~) of L-(N methyl)-alanyl-D-isoglutamine-y-tert.-butyl ester hydrochloride are obtained.
m.p. 210-12 , ~]~ = ~5 tc = 986; methanol), R = 0.15 (n-butanol/acetic acid/water = 75/7.5/21), C H ClN O calc. C 48.22 H 8.10 Cl 10.95 N 12.98 t323.82) found C 48.20 H 8010 C1 10~70 N 12.90 11.6 g t50 rnmol) of N-benzyloxycarbonyl-N-methyl-L-alanine tJ. R. Coggins and N. L. Benoiton, Canad. J~ Chem, 49, 1968 [1971]), 10.1 g t50 mmol) of D-isoglutamine-y-tert.-butyl ester and 14.8 g t60 mmol) of E~DQ are dissolved in 110 ml of dimethoxyethane and the solution is stirred at room temperature for 20 hours. The reaction solution is concentrated by evaporation, the residue is taken up in 250 ml of ethyl acetate and extracted at low temperature with 2~ 2otassium bicarbonate solution and 2~ citric acid solution. The ethyl acetate phase is washed neutral with water, deied over sodium sulphate and concentrated by eva-poration. After recrystallisation of the solid residue from dimethoxyethane/petroleum ether, 18.4 g t86 %) of N-benzyl-oxycarbonyl-L-t~-methyl)-alanyl-D-isoglutamine-y-tert.-butyl ester are obtained.
m.p. 83-85 , 835Z~3 [~ = -8 (c = 1.1; methanol), R = 0.63 (n-butanol/acetic acid/water = 75/7.5/21), C H N 0 calc. C 60.12 fl 6.96 N 10.01 (~21.49) ~ound C 60.10 H 6.90 N 9.95 Example 2: The ~ollowing are obtained in an analogous . . _ manner:
2-acetylamino-3-0-{[(D-l-carbamoyl-3-carboxy-propyl)-carbamoylmethyl]-N-methyl-carbamoylmethyl}-2-deoxy-D-glucose, [a] = +6 (c = 1, methanol), R = 0.23 (aceto~iteile/
water = 3/1), 2-benzoylamino-3-0-{[tD-l-carbamoyl-3-carboxy-propyl)-carbamoylmethyl]-N-methyl-carbamoylmethyl}-2-deoxy-D-glucose, 2-acetylamino-3 0-{[(L-l-(D-1-carbamoyl-3-carboxy-propyl)-carbamoyl-ethyl]-N-ethyl-carbamoylmethyl}-2-deoxy-D-glucose, 2-acetylamino-3-0-{[L-1-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-propyl]-N-methyl-carbamoylmethyl}-2-deoxy-D-glucose, 2-acetylamino-3-0-{[L-l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-propyl~-N-ethyl-carbamoylmethyl}-2-deoxy-D-glucose, D = -23 (c - 0.9; water), R = 0.37 (aceto-nitrile/~ater = 3/1), 2-benzoylamino-3-0-{[L-l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-propyl]~N-ethyl-carbamoylmethyl}-2-~eoxy-D-glucose, 2-acetylamino-3-0-{[L-l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-ethyl]-N-propyl-carbamoylmethyl}-2-deoxy-D-glucose, 2-acetylamino-3-0-{[L-l-(D-l-carbamoyl-3-carboxy-propyl)-N-methyl-carbamoyl-ethyl]-carbamoylmethyl}-2-~eoxy-D-glucose, 2-acetylamino-3-0-{D-l-[(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-methyl]-N-methyl-carbamoyl-ethyl}-2-deoxy-D-glucose, 2-benzoylamino-3 0-{D-l-[(D-l-c d rbamoyl-3-carboxy-propyl)-352~3 carbamoyl-methyl]-N-methyl-carbamoyl-ethyl}-2-deoxy-D-glucose, 2-acetylamino-3-0-{D-l-[L-l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-ethyl]-N-ethyl-carbamoyl-ethyl}-2-deoxy-D-glucose, 2-acetylamino-3-0-{D-l-[L-1-tD-l-caebamoyl-3-carboxy-propyl)-carbamoyl-propyl]-N-methyl~carbamoyl-etllyl}-2-deoxy-D-glucose, 2-acetylamino-3-0- f D-l-[L-l-carbamoyl-3~carboxy-propyl)-car-bamoyl-propyl~-N-ethyl-carbamoylmethyl}-2-deoxy-D-glucose, 2-benzoylamino-3-0-{D-l-[L-l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-propyl]~N-ethyl-carbamoyl-ethyl}-2-deoxy-D-glucose, 2-acetylamino-3-0-~D-l-[L-l-(D-l-carbamoyl~3-carboxy-propyl3-carbamoyl-ethyl]-N-propyl-carbamoyl-ethyl}-2-deoxy-D-glucose, 2-acetylamino-3-0-{D-l-[L-l-(D-l-carbamoyl-3-carboxy-propyl)-W-methyl-carbamoyl-ethyl]-carbamoyl-ethyl}-2-deoxy-D-glucose and 2-aaetylamino-3-0-{[L-l-(D-1,3-dicarboxy-propyl)-N-methyl-carbamoyl-ethyl]-carbamoylmethyl}-2-deoxy-D-glucose.
Example 3 A solution of 4.2 g o~ benzyl-2-(N-acetyl-N-methyl)-amino-3-0-{[L-l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-ethyl]-carbamoyl-methyl}-2-deoxy-D-glucopyranoside in 75 ml of methanol/water 1/1 is hydrogenated at 45 C and normal pressure in the presence of 0.5 g of 10 % palladium-on-carbon. The catalyst is filtered off and the filtrate is concentrated to dryness by evaporation. The residue is dissolved in a little distilled water and freeze-dried.
Thus, 2-(N-acetyl-N-methyl)-amino-3-0-{[L-l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-ethyl]-carbamoylmethyl}-2-deoxy-D-glucose is obtained in the form of a white powder.
The starting material used is manufactured as follows:
0.75 g of sodium hydride (Fluka, pract.) is added, 5Z~
while stirring and excluding moisture, under a nitrogen atmosphere, to a solution of 8.5 g of benzyl-2-acetylamino-2-deoxy-4,6-0-isopropylidene-3-0-methoxycarbonylmethyl--D-glucopyranoside in 30 ml of absolute acetonitrile and the mixture is stirred at 40 C for one hour. The reaction mixture is then cooled to room temperature and a solution of
and R represent hydrogen, the compounds of the formula I according to the invention, especially those in which the radical R represents lower alkyl, especially methyl, and more especially those in which R represents lower alkyl and R represents methyl, are distinguished by a greatly reduced pyrogenic action or, rather, one which is undetectable or tolerable in the dosages that come into consideration for practical applications, while the immuno-modulating properties remain roughly the same or are altered, usually increased, to a minor extent. These findings are of great practical significance because the pyrogenic action of the hydrophilic muramyl peptides known before the present invention in mammalsother than and as opposed ~ roden-ts is considerable and their therapeutic range is therefore too narrow to allow them to be used safe]y without const~nt medical supervision. In particular, the pyrogenic side effect could, in unfavourable circumstances, lead to thermal shock and, for that reason, certain forms of admini-stration, such as intravenous administration, are out oE
the question.
sz~
The muramyl peptides according to the invention are not toxic even when 300 mg/kg are administered i.p. to mice 5 times.
Testing for pyrogenicity was carried out on rabbits in the manner prescribed in European Pharmacopoeia, vol. 2, 1971, pages 56-59. ~ccording to this, for example even when a h gh dose, such as 10 mg/kg, of N-acetyl-muramyl-L-(~-methyl)-~-aminobutyryl-D-isoglutamine is a~ministered, no pyrogenic effect can be observed.
In the light of the state of the art, the high adju-vant activity of the compounds according to the invention in comparison with their greatly reduced pyrogenic action is surprising since, according to previous investigations, high adjuvant activity W3S always associated with a high pyrogenicity and vice versa (cf. Sh~ Kotani et al, Biken Journal 19, 9-13[1976]).
The compounds according to the invention are capable, on the one hand in admixture with an antigen, of increasing the immunogenicity of the latter and, on the other hand when administered systemically, of intensifying the immuno-logical reactivity of the treated organism. In so doing, the mentioned substances are able to promote both the cellular and the humoral immunity and to activate the lymphocytes responsible for the formation of antibodies.
The novel compounds can therefore be used as adjuvants in admixture with vaccines to improve the success of vac-cination and to improve the protection against infection mediated by humoral antibodies and/or cellular immunity against bacterial, viral or parasitic causative organisms.
Finally, the described compounds in admixture with a wide variety of antigens are suitable as adjuvants in the experimental and industrial manufacture of antisera for therapy and diagnostics and in the induction of immuno-logically activated lymphocyte populations for cell trans-fer processes.
Furthermore, the novel compounds can be used also s~
without the simultaneous administration of antigens to pro-mote immune reactions already proceeding subliminally in humans and animals. The compounds are accordingly suitable especially for the stimulation of the body's own defences, for example in chronic and acute infections or in the case of selective (antigen-speciEic) immunological defects, and in hereditary and also in acquired general (i.e. not antigen-specific) immunologically defective conditions, such as occur in old age, in the course of serious primary illnesses and especiaLly arter therapy with ionising radia-tion or with hormones having an immunosuppressive action.
The mentioned substances can therefore be administered, preferably also in combination with antibiotics admini-stered to combat infections, with chemotherapeutic agents and other pharmaceutical remedies, to counteract immunolo-gical damage. Finally, the described substances are suit-able also for the general prophylaxis of infectious diseases in humans and animals.
The present invention relates also to pharmaceutical preparations that contain compounds of the formula I. The pharmaceutical preparations according to the invention are those suitable for enteral, such as oral or rectal, admini-stration, and for parenteral administration to warm-blooded animals, which contain the pharmacological active sub-stance on its own or together with a pharmaceutically accep~
table carrier. The dosage of the active substance depends on the species of warm-blooded animal, the age and individual condition and also on the method of administra-tion and the nature of the illness treated or the nature of the desired effect.
Accordingly, the daily doses to warm-blooded animals of 70 kg body weight lie, in the case of parenteral administration, between 0.5 and 100 mg, preferably between l and 50 mg, for example 5 mg, and, in the case of oral administration, between 1 and 1500 mg, preferabLy between 5 and 750 mg, for example 50 mg~
~352~3 An appropriate fraction of the amount mentioned ~bove is administered one or more times daily, preEerably from one to three times.
The novel pharmaceutical preparations contain from approximately 10 % to approximately 95 ~, preferably from approximately 20 % to approximately 90 ~, of the active substance. Pharmaceutical preparations according to the invention can, for example, be in unit dose form, such as deagées, tablets, capsules, suppositories or ampoules.
The pharmaceutical preparations of the present invention are prepared in a manner known per se, Eor example by means of conventional mixing, granulating, coating, dis-solving or lyophilising processes~ Pharmaceutical prepara-tions for oral use can be obtained by combining the active substance with solid carriers, if desired granulating a resulting mixture and processing the mixture or granulate, if desired or necessary after the addition of suitable adjuncts, to form tablets or dragée cores.
Suitable carriers are especially fillers, such as sugars, for example :Lactose, saccharose, mannitol or sorbitol, cellulose preparations and/or calcium phosphates, for example tricalcium phosphate or calcium hydrogen phosphate, also binders, such as starch pastes, using, for example, corn, wheat, rice or potato starch, gelatin, tragacanth, methylcellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose and/or polyvinylpyrrolidone, and/or, if desired, disintegrators, such as the above-mentioned starches, also carboxymethyl starch, crosslinked polyvinylpyrrolidone, agar, alginic acid or a salt thereof, such as sodium alginate. Adjuncts are especially flow-regulators and lubricants, for example silica, talc, stearic acid or salts thereof, such as magnesium or calcium stearate, and/or polyethylene glycol. Dragée cores are provided with suitable coatings that may be resistant to gastric juice, there being used, inter alia, concentrated sugar solutions that optionally contain gum arabic, talc, polyvinylpyrrolidone, polyethylene glycol and/or titanium dioxide, lacquer solutions in suitable organic solvents or solvent mixtures or, for the manufacture of coatings that are resistant to gastric juice, solutions of suitable cellulose preparations, such as acetylcellulose phthalate or hydroxypropylmethylcellulose phthalate. Colouring sub-stances or pigments may be added to the tablets or dragée coatings, for example for identification or for indicating different doses of active substance.
In certain cases, liposomal forms of administration are especially suitable.
The following Examples illustrate the invention without limiting it in any form. The R values were ascertained on silica gel thin-layer plates manufactured by Merck. Temperatures are given in degrees Centigrade.
335;~
Example 1 7.12 g (20 mmol) of the sodium salt of benzyl-2-acetylamino-3-0-carboxymethyl-2-deoxy-4,6-0-isopropyli-dene-a-D-glucopyranoSide are suspended in 60 ml oE
dimethylformamide. After the addition of 6.46 g (20 mmol) of L-(N-methyl)-alanyl-D~isoglutamine-~-tert.-butyl ~ster hydrochloride, 5.91 g (24 mmol) of ~EDQ (2-ethoxy-N-ethox~carbonyl-1,2-dihydro-quinoline) are added to the mixture and the whole is stirred for 24 hours at room temperature. The suspension is concentrated by evaporation at 30 in vacuo, the residue is taken up in 100 ml of ethyl acetate and extracted successively, at low temperature, with 2~ potassium bicarbonate solution and 2N citric acid solution, i5 finally washed neutral with water and, after drying over sodium sulphate, concentrated by evaporation. ~he by-products still present are remoYed by chromatography over silica gel (toluene/acetone = 7/3).
Having been concentrated by evaporation, the eluate yields 7.7 9 (57 %) of henzyl-2-acetylamino-~,6-O~isopropylidene-3-O-{~L-1-(D-l~carbamoyl-3-tert.-butoxycarbonyl-propyl)-carbamoyl-ethyl]-N-methyl-carbamoylmethyl}-2-deoxy-a-D-glucopyranoside.
[~]D = +66 (c = 1.1; methanol), R = 0.81 (ethyl acetate/n-butanol/pyridine/acetic acid/
water = 42/21/21/6/lO), R = Or 56 (n~butanol/acetic acid/water = 75/7.5/21), R = ~.30 tchloroform/isopropanol/acetic acid = 70/8/2)o C H N O calc. C 58.39 1~ 7.42 ~ 8.25 33 50 ~ 11 (678.80) found C 58.01 H 7.50 N 8.20 The protecting grou?s are split off from the resulting product as follows:
7.60 g (11.1 mmol) of benzyl-2-acetylamino-4,6-0-iso-propylidene-3-0-{[L-l-(D-l-carbamoyl-3-tert.-butoxycarbonyl-propyl)-carbamoyl-ethyl]-N-methyl-carbamoylmethyl}-2-deoxy-~-D-glucopyranoside are dissolved in 80 ml of cold 95 ~ trifluoroacetic acid, the solu~ion is stirred for 15 minutes at ~ C and then 1.1 litre of ether/petroleum ether = 9/1 is added. The oily precipitate is triturated several times with fresh mixture until a solid deposit forms. The latter is dissolved in 30 ml of water and filtered over 20 ml of weakly basic ion-exchanger (Dowex-3) in acetate form to remove the trifluoroacetic acid. The column is subsequently rinsed with water and the combined eluates are lyophilised. 4.95 9 (85 %) of benzyl-2-acetylamino-3-0-{[L-l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-ethyl]-N-methyl-carbamoylmethyl}-2--deoxy-~-D-glucopyranoside are obtained.
[a]20 = +~5 (c = 1-2; H20), R - 0.28 (n-butanol/acetic acid/water - 75/7.5/21), R = 0.10 (chloroform/methanol/water=70/30/5).
4.65 g ~80 mmol) of the resulting benzyl-2-acetyl-amino-3-0-{[L-l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-ethyl]-~-methyl-carbamoylmethyl}-2-deoxy-a-D-gluco~yranoside are dissolved in 100 ml of methanol, 4.6 9 of palladium-on-carbon (10 %) are added and the mixture is treated exhaus-tively with hydrogen Eor 2 days. The catalyst is Eiltered off and the filtrate is concentrated by evaporation at room temperature. The residue is dissolved in 50 ml of water, extracted several times with n-butanol and, after evapora-ting the butanol, the aqusous phase is lyophilised. 3.2 9 (81.4 %) oE 2-acetylamino-3-0-{[L-l-(~-l-carbamoyl-3-carboxy-propyl)-carbamoyl-ethyl~-N-methyl-carbamoylmethyl}-2~deoxy-D-glucose are obtained.
[a]DO = -19 (c = 1.1; methanol), f = 0.20 (ethyl acetate/n-butanol/pyridine/acetic acid/
water = 42/21/21/6/10), ~ = 0.10 (n-butanol/acetic acid/water = 75/7.5/21), Rf = 0.45 (acetonitrile/water = 3/1) C H N 0 .1.2H 0 calc. C ~ 36 ~ 6.78 N 10.~9 19 32 ~ 11 2 (514.46) found C 44.32 H 6.75 ~ 10.79 The coupling components (muramic acid derivative and dipeptide) used as starting materials are obtained as follows:
1.2 g of sodium hydride pract~ (Fluka) is added to a solution of 8.0 g of benzyl-2-acetylamino-2-deoxy-4,6-0-iSopropylidene-a-D-glucopyranoside in 80 ml of acetonitrile and the mixture is stirred at 40 C for 2 hours. The reac-o otion mixture is subsequently cooled to from -5 to -10 and 5.2 g of bromoacetic acid methyl ester are added. The mixture is stirred for a further 30 minutes at -5 C, 10 ml of methanol are added and, after a further 10 minutes, the mixture is neutralised with acetic acid. The reaction mix-ture is concentrated to dryness by evaporation and the resi-due is partitioned between distilled water and ether. The ether solution is washed with distilled water, dried over magnesium sulphate and concentrated to dryness by evapora-tion. The resulting residue is crystallised from ether/
petroleum ether. Benzyl-2-acetylamino-3-0-methoxycarbonyl-2-deoxy-4,6-0-isopropylidene-a-D-glucopyranoside is obtained.
[~]20 = +150 ~ ~ (c = 0.875, C~C13), m.p. 122-3 C.
15 ml of lN sodium hydroxide solution are added to a solution of 4.2 g of benzyl-2-acetylamino-3-0-methoxy-carbonylmethyl-2-deoxy-4,6-0-isopropylidene-a-D~gluco-pyranoside in 20 ml of methanol and the resulting solution is left to stand at room temperature for one hour. Then, 5 ml of lN hydrochloric acid are added dropwise, while stirring, and the solution is concentrated to dryness by evaporation. The resulting po~der is dried at a high tem-perature under a high vacuum and constitutes 10 mmol of the 33~
- 2~ -~sodium salt of ben~yl-2-acetylamino-3-0-carboxy,nethyl-2-deoxy-4,6-0-isopropylidene~~-D-glucopyranoside.
18.5 g t43.9 mmol) of N-benzyloxycarbonyl-N-rllethyl-L-alanyl-D-isoglutamine-y-tert.-butyl ester are dissolved in 150 ml of methanol, 4 g of palladium-on-carbon tlO ~) are added and the mixture is hydrogenated at pE~ 4.5 tpH-stat) in the presence of 0.65N methanolic hydrochloric acid.
The catalyst is filtered off, the filtrate is concentrated by evaporation in vacuo and the residue is crystallised from 25 ml of dimethox~ethane and 100 ml of ether. 11.6 g (82 ~) of L-(N methyl)-alanyl-D-isoglutamine-y-tert.-butyl ester hydrochloride are obtained.
m.p. 210-12 , ~]~ = ~5 tc = 986; methanol), R = 0.15 (n-butanol/acetic acid/water = 75/7.5/21), C H ClN O calc. C 48.22 H 8.10 Cl 10.95 N 12.98 t323.82) found C 48.20 H 8010 C1 10~70 N 12.90 11.6 g t50 rnmol) of N-benzyloxycarbonyl-N-methyl-L-alanine tJ. R. Coggins and N. L. Benoiton, Canad. J~ Chem, 49, 1968 [1971]), 10.1 g t50 mmol) of D-isoglutamine-y-tert.-butyl ester and 14.8 g t60 mmol) of E~DQ are dissolved in 110 ml of dimethoxyethane and the solution is stirred at room temperature for 20 hours. The reaction solution is concentrated by evaporation, the residue is taken up in 250 ml of ethyl acetate and extracted at low temperature with 2~ 2otassium bicarbonate solution and 2~ citric acid solution. The ethyl acetate phase is washed neutral with water, deied over sodium sulphate and concentrated by eva-poration. After recrystallisation of the solid residue from dimethoxyethane/petroleum ether, 18.4 g t86 %) of N-benzyl-oxycarbonyl-L-t~-methyl)-alanyl-D-isoglutamine-y-tert.-butyl ester are obtained.
m.p. 83-85 , 835Z~3 [~ = -8 (c = 1.1; methanol), R = 0.63 (n-butanol/acetic acid/water = 75/7.5/21), C H N 0 calc. C 60.12 fl 6.96 N 10.01 (~21.49) ~ound C 60.10 H 6.90 N 9.95 Example 2: The ~ollowing are obtained in an analogous . . _ manner:
2-acetylamino-3-0-{[(D-l-carbamoyl-3-carboxy-propyl)-carbamoylmethyl]-N-methyl-carbamoylmethyl}-2-deoxy-D-glucose, [a] = +6 (c = 1, methanol), R = 0.23 (aceto~iteile/
water = 3/1), 2-benzoylamino-3-0-{[tD-l-carbamoyl-3-carboxy-propyl)-carbamoylmethyl]-N-methyl-carbamoylmethyl}-2-deoxy-D-glucose, 2-acetylamino-3 0-{[(L-l-(D-1-carbamoyl-3-carboxy-propyl)-carbamoyl-ethyl]-N-ethyl-carbamoylmethyl}-2-deoxy-D-glucose, 2-acetylamino-3-0-{[L-1-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-propyl]-N-methyl-carbamoylmethyl}-2-deoxy-D-glucose, 2-acetylamino-3-0-{[L-l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-propyl~-N-ethyl-carbamoylmethyl}-2-deoxy-D-glucose, D = -23 (c - 0.9; water), R = 0.37 (aceto-nitrile/~ater = 3/1), 2-benzoylamino-3-0-{[L-l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-propyl]~N-ethyl-carbamoylmethyl}-2-~eoxy-D-glucose, 2-acetylamino-3-0-{[L-l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-ethyl]-N-propyl-carbamoylmethyl}-2-deoxy-D-glucose, 2-acetylamino-3-0-{[L-l-(D-l-carbamoyl-3-carboxy-propyl)-N-methyl-carbamoyl-ethyl]-carbamoylmethyl}-2-~eoxy-D-glucose, 2-acetylamino-3-0-{D-l-[(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-methyl]-N-methyl-carbamoyl-ethyl}-2-deoxy-D-glucose, 2-benzoylamino-3 0-{D-l-[(D-l-c d rbamoyl-3-carboxy-propyl)-352~3 carbamoyl-methyl]-N-methyl-carbamoyl-ethyl}-2-deoxy-D-glucose, 2-acetylamino-3-0-{D-l-[L-l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-ethyl]-N-ethyl-carbamoyl-ethyl}-2-deoxy-D-glucose, 2-acetylamino-3-0-{D-l-[L-1-tD-l-caebamoyl-3-carboxy-propyl)-carbamoyl-propyl]-N-methyl~carbamoyl-etllyl}-2-deoxy-D-glucose, 2-acetylamino-3-0- f D-l-[L-l-carbamoyl-3~carboxy-propyl)-car-bamoyl-propyl~-N-ethyl-carbamoylmethyl}-2-deoxy-D-glucose, 2-benzoylamino-3-0-{D-l-[L-l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-propyl]~N-ethyl-carbamoyl-ethyl}-2-deoxy-D-glucose, 2-acetylamino-3-0-~D-l-[L-l-(D-l-carbamoyl~3-carboxy-propyl3-carbamoyl-ethyl]-N-propyl-carbamoyl-ethyl}-2-deoxy-D-glucose, 2-acetylamino-3-0-{D-l-[L-l-(D-l-carbamoyl-3-carboxy-propyl)-W-methyl-carbamoyl-ethyl]-carbamoyl-ethyl}-2-deoxy-D-glucose and 2-aaetylamino-3-0-{[L-l-(D-1,3-dicarboxy-propyl)-N-methyl-carbamoyl-ethyl]-carbamoylmethyl}-2-deoxy-D-glucose.
Example 3 A solution of 4.2 g o~ benzyl-2-(N-acetyl-N-methyl)-amino-3-0-{[L-l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-ethyl]-carbamoyl-methyl}-2-deoxy-D-glucopyranoside in 75 ml of methanol/water 1/1 is hydrogenated at 45 C and normal pressure in the presence of 0.5 g of 10 % palladium-on-carbon. The catalyst is filtered off and the filtrate is concentrated to dryness by evaporation. The residue is dissolved in a little distilled water and freeze-dried.
Thus, 2-(N-acetyl-N-methyl)-amino-3-0-{[L-l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-ethyl]-carbamoylmethyl}-2-deoxy-D-glucose is obtained in the form of a white powder.
The starting material used is manufactured as follows:
0.75 g of sodium hydride (Fluka, pract.) is added, 5Z~
while stirring and excluding moisture, under a nitrogen atmosphere, to a solution of 8.5 g of benzyl-2-acetylamino-2-deoxy-4,6-0-isopropylidene-3-0-methoxycarbonylmethyl--D-glucopyranoside in 30 ml of absolute acetonitrile and the mixture is stirred at 40 C for one hour. The reaction mixture is then cooled to room temperature and a solution of
6.0 g of methyl iodide in 50 ml of absolute acetonitrile is added dropwise in the course of ~ hours. AEter a further 3 hours, the reaction mixture is filtered and the filtrate is concentrated to dryness by evaporation. The residue is dissolved in ethyl acetate, the solution is washed with water, dried over sodium sulphate and concentrated to dry-ness by evaporation. Thus, benzyl-2-(N-acetyl-N-methyl)-amino-2-deoxy-4,6-0-isopropylidene-3-0-methoxycarbonyl-methyl-~-D-glucopyranoside is obtained in the form of a yellow oil.
R = 0.45 (methylene chloride/ethyl acetate = 85/15).
A solution of ~.4 g of benzyl-2-(N-acetyl-N-methyl)-amino-2-deoxy-4,6-0-isopropylidene-3-0-methoxycarbonyl-methyl-a-D-glucopyranoside in 60 ml of methanol and 15 ml of lN sodium hydroxide solution is left to stand at room temperature for one hour, 5 ml of lN hydrochloric acid are added and the mixture is concentrated to dryness by evapora-tion. The resulting sodium salt of benzyl-2-(N-acetyl-N-methyl)-amino-3-0-carboxymethyl-2-deoxy-4,6-0-isopropyli-dene-~-D-glucopyranoside is dissolved in 50 ml of N,N-dimethylformamide and condensed with 3.2 g of L-alanyl-D-isoglutamine-tert.-butyl ester hydrochloride in the presence of 2.5 g of ~EDQ. The solution is concentrated to dryness by evaporation in vacuo and the residue is dissolved in ethyl acetate. This solution is washed in succession with water, ice-cold lN hydrochloric acid, water, a saturated sodium bicarbonate solu~ion and water, is dried over mag-nesium sulphate and concentrated to dryness by evaporation.
Thus, benzyl-2-(N-acetyl-N-methyl)-amino~3-0-{[L-l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-ethyl]-carbamoyl-~i~335Z~
~ 2~ -methyl}-2-deoxy-~,6-0-isopropylidene-~-D-glucopyranoside-tert.-butyl ester is obtained in the ~orm o~ a yellowish foam. ~his product is dissolved in ~5 ml of ~5 ~ trifluoro-acetic acid that has been pre-cooled to O C and the solu-tion is stirred at O C for one hour. The reaction mix-ture is poured onto 400 ml of absolute ether, the precipi-tated product is filtered with suction, washed with ether and dried. By treating this substance with the ion-exchange resin Dowex-3 (acetate form), trifluoroacetic acid-~ree benzyl-2-(N-acetyl-N-methyl)-amino-3-0-{[L-l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-ethyl]-carbamoyl-methyl}-2-deoxy-a-D-glucopyranoside is obtained in the form of a white powder.
The following are manufactured in an analogous manner:
2-(N-acetyl-N-methyl)-amino-3-0-{D-l-[L-l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-ethyl~-carbamoyl-ethyl}-2-deoxy-D-glucose, 2-(N-acetyl-N-methyl)-amino-3-0-{D-l-[L-l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-propyl]-carbamoyl-ethyl}-2-deoxy-~-glucose, 2-(N-acetyl-N-methyl~-amino-3-0-{[L-l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-propyl]-carbamoylmethyl}-2-deoxy-D-glucose, 2-(N-acetyl-N-methyl)-amino-3-0-{[L-l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-2-methyl-propyl]-carbamoylmethyl}-2-deoxy-D-glucose, 2-(N-acetyl-N-methyl)-amino 3-0-{D-l-[L-l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-2'~methyl-propyl]-carbamoyl-ethyl}-2-deoxy-D-glucose, 3-0-{[L-l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-ethyl]-carbamoylmethyl}-2-deoxy 2-(N-propionyl-N-methyl)-amino-D-glucose, 2-(~-butyryl-N-methyl)-amino-3-0-{[L-l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-ethyl]-carbamoylmethyl}-2-deoxy-D-glucose.
R = 0.45 (methylene chloride/ethyl acetate = 85/15).
A solution of ~.4 g of benzyl-2-(N-acetyl-N-methyl)-amino-2-deoxy-4,6-0-isopropylidene-3-0-methoxycarbonyl-methyl-a-D-glucopyranoside in 60 ml of methanol and 15 ml of lN sodium hydroxide solution is left to stand at room temperature for one hour, 5 ml of lN hydrochloric acid are added and the mixture is concentrated to dryness by evapora-tion. The resulting sodium salt of benzyl-2-(N-acetyl-N-methyl)-amino-3-0-carboxymethyl-2-deoxy-4,6-0-isopropyli-dene-~-D-glucopyranoside is dissolved in 50 ml of N,N-dimethylformamide and condensed with 3.2 g of L-alanyl-D-isoglutamine-tert.-butyl ester hydrochloride in the presence of 2.5 g of ~EDQ. The solution is concentrated to dryness by evaporation in vacuo and the residue is dissolved in ethyl acetate. This solution is washed in succession with water, ice-cold lN hydrochloric acid, water, a saturated sodium bicarbonate solu~ion and water, is dried over mag-nesium sulphate and concentrated to dryness by evaporation.
Thus, benzyl-2-(N-acetyl-N-methyl)-amino~3-0-{[L-l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-ethyl]-carbamoyl-~i~335Z~
~ 2~ -methyl}-2-deoxy-~,6-0-isopropylidene-~-D-glucopyranoside-tert.-butyl ester is obtained in the ~orm o~ a yellowish foam. ~his product is dissolved in ~5 ml of ~5 ~ trifluoro-acetic acid that has been pre-cooled to O C and the solu-tion is stirred at O C for one hour. The reaction mix-ture is poured onto 400 ml of absolute ether, the precipi-tated product is filtered with suction, washed with ether and dried. By treating this substance with the ion-exchange resin Dowex-3 (acetate form), trifluoroacetic acid-~ree benzyl-2-(N-acetyl-N-methyl)-amino-3-0-{[L-l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-ethyl]-carbamoyl-methyl}-2-deoxy-a-D-glucopyranoside is obtained in the form of a white powder.
The following are manufactured in an analogous manner:
2-(N-acetyl-N-methyl)-amino-3-0-{D-l-[L-l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-ethyl~-carbamoyl-ethyl}-2-deoxy-D-glucose, 2-(N-acetyl-N-methyl)-amino-3-0-{D-l-[L-l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-propyl]-carbamoyl-ethyl}-2-deoxy-~-glucose, 2-(N-acetyl-N-methyl~-amino-3-0-{[L-l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-propyl]-carbamoylmethyl}-2-deoxy-D-glucose, 2-(N-acetyl-N-methyl)-amino-3-0-{[L-l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-2-methyl-propyl]-carbamoylmethyl}-2-deoxy-D-glucose, 2-(N-acetyl-N-methyl)-amino 3-0-{D-l-[L-l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-2'~methyl-propyl]-carbamoyl-ethyl}-2-deoxy-D-glucose, 3-0-{[L-l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-ethyl]-carbamoylmethyl}-2-deoxy 2-(N-propionyl-N-methyl)-amino-D-glucose, 2-(~-butyryl-N-methyl)-amino-3-0-{[L-l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-ethyl]-carbamoylmethyl}-2-deoxy-D-glucose.
Claims (14)
1. Process for the manufacture of compounds of the formula (I) in which X represents a carbonyl group, R1 represents hydrogen, alkyl, optionally substituted benzyl or acyl, R2 represents optionally substituted alkyl or carbocyclic aryl, R4 and R6 represent, independently of each other, hydrogen, alkyl, optionally substituted benzyl or acyl, R3 represents hydrogen or alkyl, at least one of the radicals R7, R9 and R13 represents lower alkyl, and the others represent hydrogen, R8 represents hydrogen, lower alkyl, free, esterified or etherified hydroxy-lower alkyl, free, esterified or etherified mercapto-lower alkyl, free or acylated amino-lower alkyl, cycloalkyl having 5 or 6 carbon atoms, cyclo-alkyl-lower alkyl in which the cycloalkyl radical contains 5 or 6 carbon atoms, optionally substituted aryl or aralkyl, nitrogen-containing heterocyclyl or hetero-cyclyl-lower alkyl, R7 and R8 together also represent alkylene having 3 or 4 carbon atoms, and the radicals R10, R11 and R12 represent, independently of one another, an optionally esterified or an1idated carboxy radical or R11 also represents hydrogen, or of salts of such compounds having salt-forming groups, characterised in that, a compound of the formula (II) in which X, R2, R3 and R13 have the meanings given above and R?, R? and R? represent the radicals R1, R4 and R6, respectively, or represent a protecting group that can be split off readily, or a derivative thereof, is condensed, in a manner known per se, with a compound of the formula (III) in which R7 and R9 have the meanings given above, R?, R?0, R?1 and R?2 have the meanings of R8, R10, R11 and R12, with the proviso that carboxy groups and, if desired, free hydroxyl groups present in these radicals are protected by protecting groups that can be split off readily, and optionally present protecting groups are split off, or a compound of the formula IV
(IV) in which the substituents have the meanings given above, is condensed, in a manner known per se, with a compound of the formula (V) in which R9, R?0, R?1 and R?2 have the meanings given above, with the proviso that carboxyl groups present in the radicals R?0, R?1 and R?2 and, if desired, free hydroxy groups are protected by protecting groups that can be split off readily, and optionally present protecting groups are split off, or a compound of the formula (VI) in which X, R2, R?, R?, R? and R13 have the meanings given above and hydroxy groups optionally present therein are protected by a protecting group that can be split off readily, is reacted with a compound of the formula (VII) in which Z represents a reactive esterified hydroxy group and the other substituents have the meanings given above, and optionally present protecting groups are split off and, if desired, a resulting compound of the formula I is converted into a salt.
(IV) in which the substituents have the meanings given above, is condensed, in a manner known per se, with a compound of the formula (V) in which R9, R?0, R?1 and R?2 have the meanings given above, with the proviso that carboxyl groups present in the radicals R?0, R?1 and R?2 and, if desired, free hydroxy groups are protected by protecting groups that can be split off readily, and optionally present protecting groups are split off, or a compound of the formula (VI) in which X, R2, R?, R?, R? and R13 have the meanings given above and hydroxy groups optionally present therein are protected by a protecting group that can be split off readily, is reacted with a compound of the formula (VII) in which Z represents a reactive esterified hydroxy group and the other substituents have the meanings given above, and optionally present protecting groups are split off and, if desired, a resulting compound of the formula I is converted into a salt.
2. Process according to claim 1 for the manufacture of com-pounds of the formula I, in which at least one of the radicals R7, R9 and R13 represents methyl, or salts thereof.
3. Process according to claim 1 or 2 for the manufacture of compounds of the formula I, in which R1, R4 and R6 represent hydrogen, X represents carbonyl, R2 represents lower alkyl optionally substituted by hydroxy or by methoxy, or phenyl optionally substituted by hydroxy, methoxy, methyl, ethyl or by halogen, R3 and R9 represent hydrogen or methyl, R8 represents methyl, ethyl, n-propyl, isopropyl, 2-methyl-propyl, methylmercaptomethyl, hydroxymethyl, hydroxyethyl, phenyl, benzyl or p-hydroxybenzyl, R10, R11 and R12 repre-sent carboxy, lower alkoxycarbonyl or carbamoyl or R11 also represents hydrogen or salts thereof.
4. Process according to claim 1 for the manufacture of compounds of the formula I, in which R1, R4, R6, R11 and R13 represent hydrogen, R2 represents methyl or phenyl, R3 represents hydrogen or methyl, R7 represents methyl, ethyl or propyl, R8 represents hydrogen, methyl or ethyl, R9 represents hydrogen or methyl, R10 represents carbamoyl and R12 represents carboxy, or salts thereof.
5. Process according to claim 1 or 2 for the manufacture of compounds of the formula I, in which R7 represents lower alkyl and R13 represents hydrogen, or salts thereof.
6. Process according to claim 1 for the manufacture of compounds of the formula I, in which R1, R4 and R6 repre-sent hydrogen, X represents carbonyl, R2 represents lower alkyl optionally substituted by hydroxy or by methoxy, or phenyl optionally substituted by hydroxy, methoxy, methyl, ethyl or by halogen, R3 and R9 represent hydrogen or methyl, R7 represents lower alkyl, R8 represents methyl, ethyl, n-propyl, isopropyl, 2-methylpropyl, methylmercapto-methyl, hydroxymethyl, hydroxyethyl, phenyl, benzyl or p-hydroxybenzyl, R10, R11 and R12 represent carboxy, lower alkoxycarbonyl or carbamoyl or R11 also represents hydrogen and R13 represents hydrogen, or salts thereof.
7. Process according to claim 1 for the manufacture of compounds of the formula I wherein R13 represents hydrogen or salts thereof.
8. Process according to claim 2, 4 or 6 for the manufacture of compounds of the formula I wherein R13 represents hydro-gen, or salts thereof.
9. Process according to claim 1, 2 or 4 for the manufactu-re of compounds of the formula I, wherein R3 represents methyl, or salts thereof.
10. Process according to claim 7 for the manufacture of N-acetyl-normuramyl-L-(N-methyl)-alanyl-D-isoglutamine ?
2-acetylamino-3-0- {[L-l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-ethyl]-N-methyl-carbamoylmethyl} -2-deoxy-D-glucose or salts thereof.
2-acetylamino-3-0- {[L-l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-ethyl]-N-methyl-carbamoylmethyl} -2-deoxy-D-glucose or salts thereof.
11. Process according to claim 1 for the manufacture of N-benzoyl-normuramyl-L-(N-ethyl)-.alpha.-amino-butyryl-D-iso-glutamine ? 2-benzoylamino-3-0- {[L-l-(D-l-carbamoyl-3-earboxy-propyl)-carbamoyl-propyl]-N-ethyl-carbamoylmethyl}-2-deoxy-D-glucose or salts thereof.
12. Process according to claim 1 for the manufacture of N-aeetyl-muramyl-L-(N-methyl)-.alpha.-aminobutyryl-D-isoglutamine ? 2-acetylamino-3-0- {D-l-[L-l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-propyl]-N-methyl-carbamoyl-ethyl}-2-deoxy-D-glueose or salts thereof.
13. Process aeeording to claim 1 for the manufacture of N-acetyl-muramyl-L-[N-ethyl)-alanyl-D-isoglutamine ? 2-acetylamino-3-0- {D-l-[L-l-(D-l-carbamoyl-3-carboxy-propyl)-carbamoyl-ethyl]-N-ethyl-carbamoyl-ethyl} 2-deoxy-D-glucose or salts thereof.
14. A compound of formula I as defined in claim 1 or a salt of such a compound having salt-forming groups, whenever manufactured by the process claimed in claim l, or by an obvious chemical equivalent thereof.
Applications Claiming Priority (7)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CH203578 | 1978-02-24 | ||
| CH2035/78-0 | 1978-02-24 | ||
| CH377778 | 1978-04-07 | ||
| CH3777/78-5 | 1978-04-07 | ||
| CH539478 | 1978-05-18 | ||
| CH5394/78-0 | 1978-05-18 | ||
| CA000321992A CA1138436A (en) | 1978-02-24 | 1979-02-21 | Process for the manufacture of novel antigens |
Related Parent Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA000321992A Division CA1138436A (en) | 1978-02-24 | 1979-02-21 | Process for the manufacture of novel antigens |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA1183528A true CA1183528A (en) | 1985-03-05 |
Family
ID=27426129
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA000403054A Expired CA1183528A (en) | 1978-02-24 | 1982-05-14 | Processes for the manufacture of novel glucose derivatives |
Country Status (1)
| Country | Link |
|---|---|
| CA (1) | CA1183528A (en) |
-
1982
- 1982-05-14 CA CA000403054A patent/CA1183528A/en not_active Expired
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