FR2466455A2 - Immunostimulant and adjuvant N-acyl-peptide derivs. - produced by N-acylation of peptides from streptomyces stimulosus water-soluble extract (BE 17.4.80) - Google Patents
Immunostimulant and adjuvant N-acyl-peptide derivs. - produced by N-acylation of peptides from streptomyces stimulosus water-soluble extract (BE 17.4.80) Download PDFInfo
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- FR2466455A2 FR2466455A2 FR7924923A FR7924923A FR2466455A2 FR 2466455 A2 FR2466455 A2 FR 2466455A2 FR 7924923 A FR7924923 A FR 7924923A FR 7924923 A FR7924923 A FR 7924923A FR 2466455 A2 FR2466455 A2 FR 2466455A2
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- 108090000765 processed proteins & peptides Proteins 0.000 title abstract description 5
- 102000004196 processed proteins & peptides Human genes 0.000 title abstract description 4
- 239000002671 adjuvant Substances 0.000 title abstract 2
- 229960001438 immunostimulant agent Drugs 0.000 title abstract 2
- 239000003022 immunostimulating agent Substances 0.000 title abstract 2
- 230000003308 immunostimulating effect Effects 0.000 title abstract 2
- 241000187747 Streptomyces Species 0.000 title description 5
- 230000006181 N-acylation Effects 0.000 title 1
- 239000000568 immunological adjuvant Substances 0.000 claims abstract description 5
- 239000002253 acid Substances 0.000 claims abstract 4
- 150000001875 compounds Chemical class 0.000 claims description 25
- 125000004435 hydrogen atom Chemical group [H]* 0.000 claims description 6
- 125000001931 aliphatic group Chemical group 0.000 claims description 5
- 150000001413 amino acids Chemical class 0.000 claims description 5
- 238000000034 method Methods 0.000 claims description 5
- 125000000217 alkyl group Chemical group 0.000 claims description 3
- 125000004432 carbon atom Chemical group C* 0.000 claims description 3
- NWADXBLMWHFGGU-UHFFFAOYSA-N dodecanoic anhydride Chemical group CCCCCCCCCCCC(=O)OC(=O)CCCCCCCCCCC NWADXBLMWHFGGU-UHFFFAOYSA-N 0.000 claims description 3
- 239000008194 pharmaceutical composition Substances 0.000 claims description 3
- 230000001225 therapeutic effect Effects 0.000 claims description 2
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 claims 2
- RFHAIADPANSGSA-UHFFFAOYSA-N 2,2,7-triamino-7-oxoheptanoic acid Chemical compound NC(=O)CCCCC(N)(N)C(O)=O RFHAIADPANSGSA-UHFFFAOYSA-N 0.000 claims 1
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 claims 1
- 235000004279 alanine Nutrition 0.000 claims 1
- 235000001014 amino acid Nutrition 0.000 claims 1
- 150000008064 anhydrides Chemical group 0.000 claims 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 claims 1
- 235000013922 glutamic acid Nutrition 0.000 claims 1
- 239000004220 glutamic acid Substances 0.000 claims 1
- 238000002560 therapeutic procedure Methods 0.000 claims 1
- 239000000203 mixture Substances 0.000 abstract description 13
- 239000000725 suspension Substances 0.000 abstract description 2
- 230000000240 adjuvant effect Effects 0.000 abstract 1
- 125000003342 alkenyl group Chemical group 0.000 abstract 1
- 239000013566 allergen Substances 0.000 abstract 1
- -1 amine acids Chemical class 0.000 abstract 1
- 230000000259 anti-tumor effect Effects 0.000 abstract 1
- 238000006243 chemical reaction Methods 0.000 abstract 1
- 230000003111 delayed effect Effects 0.000 abstract 1
- 238000000586 desensitisation Methods 0.000 abstract 1
- 238000009169 immunotherapy Methods 0.000 abstract 1
- 208000015181 infectious disease Diseases 0.000 abstract 1
- 229960005486 vaccine Drugs 0.000 abstract 1
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 9
- POULHZVOKOAJMA-UHFFFAOYSA-N dodecanoic acid Chemical compound CCCCCCCCCCCC(O)=O POULHZVOKOAJMA-UHFFFAOYSA-N 0.000 description 6
- DKGAVHZHDRPRBM-UHFFFAOYSA-N Tert-Butanol Chemical compound CC(C)(C)O DKGAVHZHDRPRBM-UHFFFAOYSA-N 0.000 description 5
- 239000000470 constituent Substances 0.000 description 4
- 238000001914 filtration Methods 0.000 description 4
- 239000012528 membrane Substances 0.000 description 4
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 4
- 239000002904 solvent Substances 0.000 description 4
- 239000005639 Lauric acid Substances 0.000 description 3
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 description 3
- 230000008878 coupling Effects 0.000 description 3
- 238000010168 coupling process Methods 0.000 description 3
- 238000005859 coupling reaction Methods 0.000 description 3
- 230000007062 hydrolysis Effects 0.000 description 3
- 238000006460 hydrolysis reaction Methods 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 description 2
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 238000004587 chromatography analysis Methods 0.000 description 2
- 235000019253 formic acid Nutrition 0.000 description 2
- 239000002245 particle Substances 0.000 description 2
- 239000011148 porous material Substances 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 239000002244 precipitate Substances 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- ZQXSFZAMFNRZOQ-UHFFFAOYSA-N 2-methylpropan-2-ol;hydrate Chemical compound O.CC(C)(C)O ZQXSFZAMFNRZOQ-UHFFFAOYSA-N 0.000 description 1
- BPSNETAIJADFTO-UHFFFAOYSA-N 2-pyridinylacetic acid Chemical compound OC(=O)CC1=CC=CC=N1 BPSNETAIJADFTO-UHFFFAOYSA-N 0.000 description 1
- 125000001429 N-terminal alpha-amino-acid group Chemical group 0.000 description 1
- PQLVXDKIJBQVDF-UHFFFAOYSA-N acetic acid;hydrate Chemical compound O.CC(O)=O PQLVXDKIJBQVDF-UHFFFAOYSA-N 0.000 description 1
- 238000005903 acid hydrolysis reaction Methods 0.000 description 1
- 239000008186 active pharmaceutical agent Substances 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- SIHHLZPXQLFPMC-UHFFFAOYSA-N chloroform;methanol;hydrate Chemical compound O.OC.ClC(Cl)Cl SIHHLZPXQLFPMC-UHFFFAOYSA-N 0.000 description 1
- 230000000295 complement effect Effects 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 239000012156 elution solvent Substances 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 230000008020 evaporation Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000000194 fatty acid Substances 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 150000004665 fatty acids Chemical class 0.000 description 1
- 230000003136 immunomodifying effect Effects 0.000 description 1
- QSHDDOUJBYECFT-UHFFFAOYSA-N mercury Chemical compound [Hg] QSHDDOUJBYECFT-UHFFFAOYSA-N 0.000 description 1
- 229910052753 mercury Inorganic materials 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 229920001467 poly(styrenesulfonates) Polymers 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 230000000717 retained effect Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000000377 silicon dioxide Substances 0.000 description 1
- 229910000029 sodium carbonate Inorganic materials 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
- C07K5/02—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing at least one abnormal peptide link
- C07K5/0215—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing at least one abnormal peptide link containing natural amino acids, forming a peptide bond via their side chain functional group, e.g. epsilon-Lys, gamma-Glu
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
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- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Peptides Or Proteins (AREA)
Abstract
Description
La présente addition concerne une invention rattachée à l'objet du brevet principal(brevet FR 78;29 838). The present addition relates to an invention related to the subject of the main patent (patent FR 78; 29,838).
Le brevet principal est relatif à des composés hydrosolubles obtenus par couplage d'un extrait hydrosoluble de Streptomyces stimulosus 25556 (NRRL 5776) avec des acides aliphatiques à longues chaines,lesdits composés se présentant sous la forme d'un mélange dont les constituants sont présents dans un rapport molaire voisin de 4/1 et sont représentés respectivement par les formules générales suivantes:
dans lesquelles X représente un radical R-CO- et Y représente un atome d'hydrogène ou un radical RCO,ou bien dans lesquelles
Y représente un radical RCO et X représente un atome d'hy drogène ou un radical RCO,étant entendu que,dans ces définitions, R représente un radical alcoyle ou alcényle contenant 11 à 19 atomes de carbone en chaîne droite ou ramifiée et dans lesquelles les acides aminés ont les configurations suivantes:
Ala: L,Glu :D et DAP : L, L. The main patent relates to water-soluble compounds obtained by coupling a water-soluble extract of Streptomyces stimulosus 25556 (NRRL 5776) with long-chain aliphatic acids, said compounds being in the form of a mixture, the constituents of which are present in a molar ratio close to 4/1 and are represented respectively by the following general formulas:
in which X represents an R-CO- radical and Y represents a hydrogen atom or an RCO radical, or in which
Y represents an RCO radical and X represents a hydrogen atom or an RCO radical, it being understood that, in these definitions, R represents an alkyl or alkenyl radical containing 11 to 19 carbon atoms in a straight or branched chain and in which the amino acids have the following configurations:
Ala: L, Glu: D and DAP: L, L.
Ces composés sont des adjuvants immunologiques particulièrement intéressants. These compounds are particularly advantageous immunological adjuvants.
Lesdits composés hydrosolubles proviennent du couplage avec des acides aliphatiques à longue chaîne d'un mélange de deux constituants de formules (I) et (II) dans un rapport molaire de 4/1 environ:
Le mélange des deux constituants de formules (I) et (II) dans le rapport molaire d'environ 4/l,désigné au brevet principal par le numéro 39592 RP peut être obtenu par purifications successives d'extrait,désigné sous le numéro 34129 RP,de Streptomyces stimulosus DS 25556 (NRRL 5776) par chromatographie sur divers supports appropriés,tels que les "Biogels". The mixture of the two constituents of formulas (I) and (II) in the molar ratio of approximately 4 / l, designated in the main patent by the number 39592 RP can be obtained by successive purifications of extract, designated under the number 34129 RP , of Streptomyces stimulosus DS 25556 (NRRL 5776) by chromatography on various suitable supports, such as "Biogels".
I1 a maintenant été trouvé,et c'est ce qui fait l'un des objets de la présente addition,que le mélange des produits de formule (I) et (II) peut être séparé en ses constituants
La présente addition concerne donc le tétrapeptide de formule:
et le tétrapeptide de formule:
leur séparation à partir d'un extrait de Streptomyces stimulosus ainsi que leur utilisation pour la préparation d'adjuvants immunologiques.It has now been found, and this is one of the objects of the present addition, that the mixture of the products of formula (I) and (II) can be separated into its constituents
The present addition therefore relates to the tetrapeptide of formula:
and the tetrapeptide of formula:
their separation from an extract of Streptomyces stimulosus as well as their use for the preparation of immunological adjuvants.
Selon la présente additionXles tétrapeptides de formules (I) et (II) peuvent être obtenus en soumettant le mélange 39592 RP à une chromatographie sur une colonne Dowex 50 x 4 en éluant avec un mélange de solvants convenables dont l'acidité va en décroissant. Les tétrapeptides (I) et (II) sont élués de la colonne pour une valeur déterminée du pH qui dépend du système de solvants utilisés. According to the present addition, the tetrapeptides of formulas (I) and (II) can be obtained by subjecting the mixture 39592 RP to chromatography on a Dowex 50 × 4 column, eluting with a mixture of suitable solvents whose acidity is decreasing. The tetrapeptides (I) and (II) are eluted from the column for a determined value of the pH which depends on the system of solvents used.
Comme solvant d'élution,il est particulièrement intéressant d'utiliser un mélange pyridine-acide acétiqueeau dont le pH est ajusté à la valeur désirée par addition d'acide formique. Dans un tel système le tétrapeptide de formule (I) est élué lorsque le pH atteint 4,1. As an elution solvent, it is particularly advantageous to use a pyridine-acetic acid water mixture, the pH of which is adjusted to the desired value by addition of formic acid. In such a system the tetrapeptide of formula (I) is eluted when the pH reaches 4.1.
Chacun des tétrapeptides ainsi obtenus est désalifié par filtration sur une colonne de Biogel P2 en éluant avec de l'acide acétique 0,01 M. Each of the tetrapeptides thus obtained is desalted by filtration on a column of Biogel P2, eluting with 0.01 M acetic acid.
La structure des tétrapeptides de formules (I) et (II) a été déterminée par -leur composition en aminoacides: Glu (1,0), Gly (1,0),
Ala (1,0) et DAP (1,0).The structure of the tetrapeptides of formulas (I) and (II) was determined by their amino acid composition: Glu (1.0), Gly (1.0),
Ala (1.0) and DAP (1.0).
-la détermination des aminoacides N-terminaux par les méthodes classiques d'Edman et de dansylation et aussi par hydrolyse acide partielle.the determination of N-terminal amino acids by the conventional Edman and dansylation methods and also by partial acid hydrolysis.
Les tétrapeptides de formules (I) et (II) peuvent être respectivement couplés avec les acides gras en opérant dans les conditions décrites dans le brevet principal. The tetrapeptides of formulas (I) and (II) can be respectively coupled with the fatty acids by operating under the conditions described in the main patent.
Ainsi, sous un aspect complémentaire,la présente invention concerne également les produits de couplage des tétrapeptides de formules (I) et (II), à savoir d'une part le composé de formule (III)
et d'autre part le composé de formule (IV)
formules dans lesquelles X représente un radical R-CO- et
Y représente un atome d'hydrogène ou un radical RCO, ou bien dans lesquelles Y représente un radical RCO et X représente un atome d'hydrogène ou un radical RCO, étant entendu que, dans ces définitions, R représente un radical alcoyle ou alcényle contenant 11 à 19 atomes de carbone en chai ne droite ou ramifiée et dans lesquelles les acides aminés ont les configurations suivantes: Ala: L, Glu: D, et DAP: L,L.Thus, in a complementary aspect, the present invention also relates to the coupling products of the tetrapeptides of formulas (I) and (II), namely on the one hand the compound of formula (III)
and on the other hand the compound of formula (IV)
formulas in which X represents an R-CO- radical and
Y represents a hydrogen atom or an RCO radical, or else in which Y represents an RCO radical and X represents a hydrogen atom or an RCO radical, it being understood that, in these definitions, R represents an alkyl or alkenyl radical containing 11 to 19 carbon atoms in a straight or branched chain, in which the amino acids have the following configurations: Ala: L, Glu: D, and DAP: L, L.
Sous une forme préférées l'invention a pour objet le composé de formule (III). In a preferred form the invention relates to the compound of formula (III).
On notera que dans la formule (III) les symboles
X et Y peuvent être identiques ou différents, de sorte que le composé de formule (III) peut se présenter sous la forme d'un composé unique ou d'un mélange de composés. Dans la présente description, l'expression "composé de formule (III)" couvre ces diverses possibilités.Note that in formula (III) the symbols
X and Y may be the same or different, so that the compound of formula (III) may be in the form of a single compound or a mixture of compounds. In the present description, the expression "compound of formula (III)" covers these various possibilities.
L'exemple suivant,donné à titre non limitatif,illustre la présente addition. The following example, given without limitation, illustrates the present addition.
EXEMPLE
a. Preparation du tétrapeptide (I)
30 g de cellules délipidées de Streptomyces stimulosus sont mis en suspension dans 300 cm3 d'eau contenant 0,5 cm3 de toluène. Lo suspension est agitée pendant 48 heures à 370C puis elle est centrifugée pendant 30 minutes à 15000 g.EXAMPLE
at. Preparation of tetrapeptide (I)
30 g of defatted cells of Streptomyces stimulosus are suspended in 300 cm3 of water containing 0.5 cm3 of toluene. Lo suspension is stirred for 48 hours at 370C then it is centrifuged for 30 minutes at 15000 g.
Le surnageant est filtré sur une membrane AMICON
UM-20 puis sur membrane AMICON UM-2. Les substances hydrosolubles qui sont retenues par la membrane UM-2 sont purifiées par filtrations successives sur des colonnes (240 cm x 1,2 cm) de Biogel P 10, P 4 et P 2 en éluant avec 240 cm3 d'acide acétique 0,01 M.The supernatant is filtered through an AMICON membrane
UM-20 then on AMICON UM-2 membrane. The water-soluble substances which are retained by the UM-2 membrane are purified by successive filtrations on columns (240 cm x 1.2 cm) of Biogel P 10, P 4 and P 2, eluting with 240 cm3 of acetic acid 0, 01 M.
Les peptides (15 mg) ainsi obtenus après filtration sur Biogel P 2 sont chromatographiés sur une colonne Doiex 50 x 4 ( 100 cm x 1,4 cm) tamponnée au départ à pH 2,2 avec un solvant pyridine:acide acétique-eau (16:130:854 en volumes)ajusté au pH désiré avec de l'acide formique. Avec un gradient d'élution de 500 cm3,le pH étant alors de 4,1, le tétrapeptide de formule (I) est obtenu.Il est purifié par filtration sur une colonne de Biogel P2 en éluant avec 240cm3 d'acide acétique 0,01M. The peptides (15 mg) thus obtained after filtration on Biogel P 2 are chromatographed on a Doiex 50 x 4 column (100 cm x 1.4 cm) buffered at the start at pH 2.2 with a pyridine solvent: acetic acid-water ( 16: 130: 854 by volume) adjusted to the desired pH with formic acid. With an elution gradient of 500 cm3, the pH then being 4.1, the tetrapeptide of formula (I) is obtained. It is purified by filtration on a column of Biogel P2, eluting with 240cm3 of acetic acid 0, 01M.
On obtient ainsi 7,5mg de tétrapeptide de formule(I). 7.5 mg of tetrapeptide of formula (I) are thus obtained.
b . Préparation dutomposé de formule (III)
(RCO=radical lauroyle)
On dissout 15 mg du produit de formule (I) dans 5 cm3 d'un mélange d'une solution aqueuse 0,5 M de carbonate de sodium et d'alcool butylique tertiaire (7/3 en volumes).b. Preparation of the compound of formula (III)
(RCO = lauroyl radical)
15 mg of the product of formula (I) are dissolved in 5 cm 3 of a mixture of a 0.5 M aqueous solution of sodium carbonate and tertiary butyl alcohol (7/3 by volume).
On ajoute goutte à goutte,sous agitation et à une température voisine de 200C, 0,5 cm3 d'alcool butyllque tertiaire contenant 60 mg d'anhydride laurique. Après avoir agité pendant 30 minutes,on ajoute de l'alcool butylique tertiaire jusqu'à obtention d'un taux d'alcool butylique tertiaire de 80%, I1 se forme alors un précipité que l'on sépare par centrifugation (3000 tours/minute pendant 10 minutes). Le précipité est lavé 2 fois par 10 cm3 d'alcool butylique tertiaire et centrifugé à chaque fois dans les mêmes conditions que cidessus. Tous les surnageants de centrifugation sont réunis et concentrés à sec à l'évaporateur rotatif sous pression réduite (20 mm de mercure)à une température inférieure à 300C.Le résidu d'évaporation est repris par 50 cm3 d'eau distillée et dialysé à travers une membrane Diaflo (AMICON)
UM-05,afin d'éliminer la majorité de l'acide laurique provenant de l'hydrolyse de l'anhydride laurique n'ayant pas réagi. La solution qui n'a pas dialysé est lyophilisée.0.5 cm3 of tertiary butyl alcohol containing 60 mg of lauric anhydride is added dropwise, with stirring and at a temperature in the region of 200C. After stirring for 30 minutes, tertiary butyl alcohol is added until a tertiary butyl alcohol level of 80% is obtained, a precipitate is formed which is separated by centrifugation (3000 revolutions / minute for 10 minutes). The precipitate is washed twice with 10 cm 3 of tertiary butyl alcohol and centrifuged each time under the same conditions as above. All the centrifugation supernatants are combined and concentrated to dryness on a rotary evaporator under reduced pressure (20 mm of mercury) at a temperature below 300C. The evaporation residue is taken up in 50 cm3 of distilled water and dialyzed through a Diaflo membrane (AMICON)
UM-05, to remove the majority of lauric acid from the hydrolysis of unreacted lauric anhydride. The solution which has not dialyzed is lyophilized.
On obtient ainsi 30 mg d'une poudre blanche amorphe contenant encore 10% d'acide laurique,30 mg de ladite poudre sont filtrés sur Spherosil XOA-400 (qui est une résine à base de silice en particules sphériques poreuses de granulométrie comprise entre 100 et 300 microns,dont le volume poreux est de 1 ml/g et le diamètre des pores voisin de 80 angströms mise sur le marché par Rhône-Poulenc Industries) dans une colonne de 25 x 1 cm dans le solvant chloroforme-méthanol-eau (75/25/2). L'acide laurique est élué dans le volume mort de la colonne et le composé (III) est élué par le mélange butanol tertiaire-eau (50/50). La composition relative des acides aminés dans le composé (III),déterminée par un autoanalyseur après hydrolyse totale,est la suivante (entre parenthèses est indiqué le nombre entier le plus proche en accord avec des expériences effectuées après différents temps d'hydrolyse):
Ala= 1,08 (1); Glu: 0,79(1);Gly=1,0 (l);Dap=0,83 (1).30 mg of an amorphous white powder are thus obtained, still containing 10% of lauric acid, 30 mg of said powder are filtered through Spherosil XOA-400 (which is a silica-based resin in spherical porous particles with a particle size between 100 and 300 microns, whose pore volume is 1 ml / g and the diameter of the pores close to 80 angstroms placed on the market by Rhône-Poulenc Industries) in a column of 25 x 1 cm in the chloroform-methanol-water solvent ( 75/25/2). The lauric acid is eluted in the dead volume of the column and the compound (III) is eluted by the tertiary butanol-water mixture (50/50). The relative composition of the amino acids in compound (III), determined by an autoanalyzer after total hydrolysis, is as follows (in brackets the closest whole number is indicated in accordance with experiments carried out after different hydrolysis times):
Ala = 1.08 (1); Glu: 0.79 (1); Gly = 1.0 (l); Dap = 0.83 (1).
Le composé de formule (III) possède des propriétés d'immunomodification. Les indications concernant ses applications thérapeutiques et ses propriétés pharmacologiques sont les mêmes que celles décrites au brevet principal. The compound of formula (III) has immunomodifying properties. The indications concerning its therapeutic applications and its pharmacological properties are the same as those described in the main patent.
I1 suffira donc de se reporter,si besoin est,à la description du brevet principal,pour tout ce qui concerne l'utilité du composé (III) comme médicament ainsi que celle des compositions pharmaceutiques le contenant. It will therefore suffice to refer, if necessary, to the description of the main patent, for all that relates to the usefulness of the compound (III) as a medicament as well as that of the pharmaceutical compositions containing it.
Claims (9)
Priority Applications (7)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| FR7924923A FR2466455A2 (en) | 1979-10-05 | 1979-10-05 | Immunostimulant and adjuvant N-acyl-peptide derivs. - produced by N-acylation of peptides from streptomyces stimulosus water-soluble extract (BE 17.4.80) |
| GB7935476A GB2033906B (en) | 1978-10-19 | 1979-10-12 | Water-soluble compounds derived from extracts of streptomyces stimulosus process for their production and compositions containing them |
| DE19792941551 DE2941551A1 (en) | 1978-10-19 | 1979-10-13 | IMMUNOLOGICALLY EFFECTIVE COMPOUNDS, METHOD FOR THEIR PRODUCTION AND MEDICINAL PRODUCTS CONTAINING THE SAME |
| US06/084,927 US4261979A (en) | 1978-10-19 | 1979-10-15 | Novel water-soluble compounds derived from extracts of streptomyces stimulosus, process for their production, their uses as immunological adjuvants and compositions containing them |
| NL7907637A NL7907637A (en) | 1978-10-19 | 1979-10-16 | NEW WATER-SOLUBLE COMPOUNDS FROM A STREPTOMYCES STIMULOSUS EXTRACT, METHODS FOR OBTAINING THEM, THEIR USE AS IMMUNOLOGICAL ADDITIVES AND PREPARATIONS CONTAINING THE NEW COMPOUNDS. |
| CH937879A CH643816A5 (en) | 1978-10-19 | 1979-10-18 | WATER-SOLUBLE COMPOUNDS FROM STREPTOMYCES STIMULOSUS EXTRACT, THEIR PRODUCTION, THEIR USE AS IMMUNOLOGICAL ADJUVANTS AND COMPOSITIONS CONTAINING THEM. |
| CA000337927A CA1149376A (en) | 1978-10-19 | 1979-10-18 | Process for preparing water-soluble compounds derived from extracts of streptomyces stimulosus, and water-soluble compounds so obtained |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| FR7924923A FR2466455A2 (en) | 1979-10-05 | 1979-10-05 | Immunostimulant and adjuvant N-acyl-peptide derivs. - produced by N-acylation of peptides from streptomyces stimulosus water-soluble extract (BE 17.4.80) |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| FR2466455A2 true FR2466455A2 (en) | 1981-04-10 |
| FR2466455B2 FR2466455B2 (en) | 1983-12-23 |
Family
ID=9230402
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| FR7924923A Granted FR2466455A2 (en) | 1978-10-19 | 1979-10-05 | Immunostimulant and adjuvant N-acyl-peptide derivs. - produced by N-acylation of peptides from streptomyces stimulosus water-soluble extract (BE 17.4.80) |
Country Status (1)
| Country | Link |
|---|---|
| FR (1) | FR2466455A2 (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| FR2248025A2 (en) * | 1973-10-23 | 1975-05-16 | Anvar | Vaccine compsns. - contg. N-acyl muramic acid deriv. as adjuvant |
| FR2292486A1 (en) * | 1974-07-01 | 1976-06-25 | Anvar | WATER-SOLUBLE IMMUNOLOGICAL ADJUVANTS, ESPECIALLY FOR VACCINES, OBTAINED FROM MICROBACTERIA AND OTHER MICRO-ORGANISMS |
| FR2399437A1 (en) * | 1977-08-04 | 1979-03-02 | Anvar | NEW IMMUNOSTIMULANT SUBSTANCE, ITS PREPARATION PROCESS AND THE COMPOSITIONS CONTAINING IT |
-
1979
- 1979-10-05 FR FR7924923A patent/FR2466455A2/en active Granted
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| FR2248025A2 (en) * | 1973-10-23 | 1975-05-16 | Anvar | Vaccine compsns. - contg. N-acyl muramic acid deriv. as adjuvant |
| FR2292486A1 (en) * | 1974-07-01 | 1976-06-25 | Anvar | WATER-SOLUBLE IMMUNOLOGICAL ADJUVANTS, ESPECIALLY FOR VACCINES, OBTAINED FROM MICROBACTERIA AND OTHER MICRO-ORGANISMS |
| FR2399437A1 (en) * | 1977-08-04 | 1979-03-02 | Anvar | NEW IMMUNOSTIMULANT SUBSTANCE, ITS PREPARATION PROCESS AND THE COMPOSITIONS CONTAINING IT |
Also Published As
| Publication number | Publication date |
|---|---|
| FR2466455B2 (en) | 1983-12-23 |
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