A subject matter of the present invention is novel glycopolymers, their uses and novel monomers of use in their preparation. The invention also relates to processes for preparation of the novel monomers and glycopolymers.
Glycopolymers are polymers comprising units comprising a glycoside unit. They can be obtained by polymerization of monomers comprising a glycoside, by copolymerization in the presence of other monomers or by grafting to a polymer functionalized for this purpose.
Numerous compounds comprising a glycoside group, generally a monoglycoside group, and a polymerizable group, for example a double bond, have been described. The polymerization of such compounds has also been described.
For example, the document WO 90/10023 discloses glycopolymers comprising units derived from acrylamide and units deriving from monomers of formula R2—NH—CO—CX═CH2 where X is H or a methyl group and R2 is a glycoside. These monomers are obtained from compounds of formula R2—NH2 in which the R2 group is bonded to the —NH2 group via a reducing anomeric carbon.
The present invention provides other glycopolymers and other monomers comprising a glycoside. These novel glycopolymers and monomers can be of use in adjusting the properties of the glycopolymers and can thus make it possible, when they are used, to provide novel products. It is thus possible to adjust the properties of polymers used, for example, in cosmetic compositions.
Furthermore, glycopolymers are attracting increasing interest in the industrial and/or consumables fields as they are products derived from natural products, benefiting from a positive image in terms of environmental protection and/or of harmfulness and/or more simply of marketing. There exists a need for such products.
Thus, the invention provides a polymer comprising units comprising a glycoside, characterized in that it comprises:
The invention also relates to uses of the polymer in compositions. The invention also relates to compositions comprising the polymer.
The invention also relates to a monomer particularly suitable for the preparation of the polymer according to the invention. Thus, the invention also provides a monomer of following formula (I′)
in which:
—COY-L1-S-L2-Z-G (II′)
The monomer according to the invention can be used for the preparation of the polymers according to the invention. It can also be used for the preparation of other polymers, for example homopolymers of said monomer, or for the preparation of copolymers not comprising cationic or potentially cationic units but comprising other units. It may involve, for example, copolymers comprising units deriving from the monomer of formula (I′) and, as other units, neutral, anionic and/or potentially anionic and hydrophobic and/or hydrophilic units. Such units are described subsequently.
The term “polymer” is understood to mean any macromolecular compound comprising repeat units. Polymers include in particular homopolymers, copolymers, oligomers, cooligomers, telomers and cotelomers.
The term “copolymer” is understood to mean any polymer comprising at least two different repeat units. Copolymers include in particular random copolymers, controlled structure copolymers, cooligomers (copolymers of relatively low molecular weight) and cotelomers.
The term “controlled structure (co)polymer” is understood to mean any (co)polymer where the sequence of the units is controlled (for example, diblock or triblock copolymers but also concentration gradient polymers) and/or where the polydispersity is controlled (for example, random (co)polymers having a polydispersity index of 1 to 1.5), in contrast to the (co)polymers obtained by standard polymerization processes, which do not make possible such a control of the arrangement of the individual units or of the polydispersity indices, if low. It may concern a copolymer comprising at least two parts A and B with distinct compositions of repeat units. The parts of a controlled structure copolymer can in particular be blocks, linear backbones, side chains, grafts, “hairs” or branches of microgels or of stars, cores of stars or of microgels, or alternatively parts of polymer chains exhibiting different concentrations of different units. Thus, the controlled structure, which a copolymer can exhibit, can be chosen from the following structures:
The term “monomers” is understood to mean compounds which can be used for the preparation of polymers, homopolymers or copolymers (it is also possible to speak of comonomers). The repeat units of the polymers derive from these monomers.
In the present patent application, the term “unit deriving from a monomer” denotes a unit which can be obtained directly from said monomer by polymerization. Thus, for example, a unit deriving from an acrylic or methacrylic acid ester does not cover a unit of formula —CH2—CH(COOH)—, —CH2—C(CH3)(COOH)— or —CH2—CH(OH)— respectively, obtained, for example, by polymerizing an acrylic acid ester, a methacrylic acid ester or vinyl acetate respectively and by then hydrolyzing. A unit deriving from acrylic or methacrylic acid covers, for example, a unit obtained by polymerizing a monomer (for example, an acrylic or methacrylic acid ester) and by then reacting (for example by hydrolysis) the polymer obtained so as to obtain units of formula —CH2—CH(COOH)— or —CH2—C(CH3)(COOH)—. A unit deriving from a vinyl alcohol covers, for example, a unit obtained by polymerizing a monomer (for example, a vinyl ester), and by then reacting (for example by hydrolysis) the polymer obtained so as to obtain units of formula —CH2—CH(OH)—.
In the present patent application, unless otherwise mentioned, the average molar masses are absolute weight-average molar masses which can be measured by steric exclusion chromatography in an appropriate solvent (for example, deionized Millipore water, if appropriate), coupled to a refractometer, to a conductivity meter and to a multi-angle light scattering detector, with extrapolation to angle zero (GPC-MALS).
In the present patent application, “Ac” represents an acetyl group of formula —COCH3.
In the present patent application, the term glycoside refers to any group comprising one or more glycoside units, and to the derivatives of these groups. In the case where the glycoside comprises several glycoside units, the term “polyglycosides” is also used. The term “polyglycoside” is understood to mean a glycoside comprising at least two glycoside units.
Glycoside units, glycosides, polyglycosides, their derivatives, their structures and formulae are known to a person skilled in the art. It is specified, for the glycoside units, that it can be a matter in particular of aldoses, of ketoses or of derivatives in rings comprising 5 atoms (pentoses) or 6 atoms (hexoses). In addition, it is known to a person skilled in the art that glycosides, polyglycosides and their derivatives exhibit a reducing “anomeric carbon” at one end, the right-hand end according to writing conventions. It is also known that glycoside units, glycosides, polyglycosides and their derivatives exhibit optionally protected hydroxyl (—OH), carboxylic acid or amine groups.
Glycosides include in particular:
Mention is made, as examples of monosaccharide glycosides, of the following glycosides;
glucose (for example D-glucose), fructose, sorbose, mannose, galactose, talose, allose, gulose, idose, glucosamine, mannoamine, galactosamine, glucuronic acid, rhamnose, arabinose, galacturonic acid, fucose, xylose, lyxose, ribose, generally the isomers of sucrose, such as palatinose.
Mention is made, as examples of di- or oligosaccharide glycosides, of the following glycosides:
Mention is also made, as examples of glycosides, of:
Polyglycosides, comprising several glycoside units, can be described as sequences of glycosides (mono- and/or polyglycosides). In the present patent application, a sequence of glycosides is described by the formula Ga-Gb-, in which Ga is a glycoside or a polyglycoside and Gb is a glycoside or a polyglycoside. In the case where Ga or Gb is a polyglycoside, the latter can also be described by a formula Ga′-Gb′-, in which Ga′ is a glycoside or a polyglycoside and Gb′ is a glycoside or a polyglycoside, and so on. Glycosides or polyglycosides which can constitute Ga, Gb, Ga′, Gb′, and the like, groups have been mentioned above.
The polymer comprises units deriving from a monomer of following formula (I):
G can be bonded to -Z- via an anomeric carbon atom or via another carbon atom.
G can in particular be bonded via:
G can also be grafted by reductive amination.
If G comprises an acid or amine functional group on other positions, it is possible to graft via this functional group.
Preferably:
According to one embodiment, the -L-Z-G group is a group of formula —O—CH2CH2—O-G. Monomers exhibiting such a group are sold, for example, by Nippon Seika under the name Sucrograph.
According to another embodiment, the -L-Z-G group can be a group of formula —CO—NH-G or —CO-aryl-NH-G.
According to a specific embodiment of the invention, the monomer of formula (I) is a monomer of formula (I′) as described below.
A monomer particularly suited to the implementation of the invention exhibits the following formula (I′)
in which:
—COY-L1-S-L2-Z-G (II′)
Thus, the compound of formula (I′) exhibits the following formula (I″):
Preferably:
According to a particular form, the monomer of formula (I) or (I′) or (I″) exhibits the following formula (III′):
In this formula:
In particular, the monomer of the formula (I), (I′), (I″) or (III″) can exhibit one of the following formulae:
in which:
m and n, which are identical or different, are numbers from 0 to 10, preferably 0 or 1.
The monomer of formula (I′) can be prepared by a process comprising the following stages:
The reaction can be carried out in the absence of solvent, at ambient temperature, but other reaction methods are not ruled out.
Thus, stage a) can comprise the following stages:
a1) reaction of the anomeric carbon of a glycoside of formula G-OH, comprising free —OH groups, with excess allylamine,
a2) removal of the excess allylamine,
a3) reaction with acetic anhydride, so as to protect the nitrogen atom and optionally primary —OH groups of the glycoside.
Stage a3) of reaction with acetic anhydride can be carried out under conditions such that at least a portion of the —OH groups of the glycoside are acetylated, in addition to the nitrogen atom. It is possible to promote this acetylation of —OH groups or to retain it or to eliminate it during a subsequent stage, for example, by a slightly basic treatment which hydrolyzes the O-acetate groups.
The addition of the compound can be carried out by radical reaction, either in the presence of a radical initiator or by photochemical reaction. In both cases, the reaction preferably takes place in a minimum amount of solvent, for example water, if need be while heating. According to an advantageous form, the reaction is carried out in an aqueous medium. According to a particular advantageous form, the reaction is carried out in an aqueous medium using a water-soluble radical initiator. This embodiment can make it possible in particular to employ smaller amounts of solvent, to increase the reaction kinetics and to improve the final yield obtained. Initiators which can be used are known to a person skilled in the art. Mention is made, by way of examples, of V50 (α,α′-azodiisobutyramidine dihydrochloride), VA-041 (2,2′-azobis[2-(5-methyl-2-imidazolin-2-yl)propane]dihydrochloride) or VA-060 (2,2′-azobis{2-[1-(2-hydroxyethyl)-2-imidazolin-2-yl]propane}dihydrochloride). One embodiment of this stage can be represented by the following reaction scheme:
For all the embodiments, the glycoside G is preferably a polyglycoside. Glycosides which can constitute G groups of monomers of formula (I), (I′), (I″), or (III′) have been described above in the “Definitions” section.
The polymer according to the invention comprises cationic or potentially cationic units AC which can derive from cationic or potentially cationic monomers.
The term “cationic or potentially cationic units AC” is understood to mean units which comprise a cationic or potentially cationic group. Cationic units or groups are units or groups which exhibit at least one positive charge (generally in combination with one or more anions, such as the chloride ion, the bromide ion, a sulfate group or a methyl sulfate group), whatever the pH of the medium in which the copolymer is present. Potentially cationic units or groups are units or groups which may be neutral or which may exhibit at least one positive charge, depending on the pH of the medium in which the copolymer is present. In this case, reference will be made to potentially cationic units AC in the neutral form or in the cationic form. By extension, it is possible to speak of cationic or potentially cationic monomers.
Mention may be made, as examples of potentially cationic hydrophilic monomers (from which units AC can derive), or:
Mention may be made, as examples of cationic hydrophilic monomers, from which units AC can be derived, of:
The polymer can also comprise other units, for example neutral hydrophilic or hydrophobic units AN and/or anionic or potentially anionic units AA.
The term “anionic or potentially anionic units AA” is understood to mean units which comprise an anionic or potentially anionic group. Anionic units or groups are units or groups which exhibit at least one negative charge (generally in combination with one or more cations, such as cations of alkali metal or alkaline earth metal compounds, for example sodium, or cationic groups, such as ammonium), whatever the pH of the medium in which the copolymer is present. Potentially anionic units or groups are units or groups which may be neutral or which may exhibit at least one negative charge, depending on the pH of the medium in which the copolymer is present. In this case, reference will be made to potentially anionic units AA in the neutral form or in the anionic form. By extension, it is possible to speak of anionic or potentially anionic monomers.
The term “neutral units AN” is understood to mean units which do not exhibit a charge, whatever the pH of the medium in which the copolymer is present.
Mention may be made, as examples of anionic or potentially anionic monomers, from which units AA can be derived, of:
Mention may be made, as examples of neutral nonionic hydrophobic monomers, from which units AN can be derived, of:
Thus, the part B can be a silicone, for example a polydimethylsiloxane chain or a copolymer comprising dimethylsiloxy units.
Mention may be made, as examples of neutral nonionic hydrophilic monomers, from which units AN can be derived, of:
The polymer according to the invention can be a random copolymer, a block copolymer, a concentration gradient copolymer, a star copolymer, a cooligomer or a cotelomer. It is preferably a random copolymer.
According to an advantageous embodiment, the polymer is water-soluble or water-dispersible. This means that said polymer does not form, in water, over at least in a certain pH and concentration range, a two-phase composition under the conditions of use.
The polymer according to the invention can be presented in particular in the form of a powder, in the form of a dispersion in a liquid or in the form of a solution in a solvent (water or other). The form depends generally on the requirements related to the use of the polymer. It can also be related to the process for the preparation of the polymer.
The polymer can comprise from 0.1% to 99.9% by number (molar) of units deriving from the monomer of formula (I) or (I′), with respect to the total number of units in the polymer. It preferably comprises from 0.1% to 15% by number (molar).
The polymer can comprise from 0.1% to 99.9% by number (molar) of cationic or potentially cationic units, with respect to the total number of units in the polymer. It preferably comprises from 0.1% to 15% by number (molar).
The absolute weight-average molar mass can preferably be between 1000 g/mol and 5 00 000 g/mol. It is preferably between 50 000 g/mol and 1 000 000 g/mol.
The polymers according to the invention can be obtained by any known method, whether by controlled or uncontrolled radical polymerization, by polymerization by ring opening (in particular anionic or cationic, with appropriate monomers), by anionic or cationic polymerization or by chemical modification of a polymer.
Radical polymerization is preferably carried out in an environment devoid of oxygen, for example in the presence of an inert gas (helium, argon, nitrogen, and the like). The reaction is carried out in an inert solvent, preferably methanol or ethanol, and more preferably in water.
The polymerization is initiated by addition of a polymerization initiator. The initiators used are the free radical generators commonly used in the art. Examples comprise organic peresters; organic compounds of azo type, for example azobisamidinopropane hydrochloride, azobisisobutyronitrile, azobis(2,4-dimethylvaleronitrile), and the like; inorganic and organic peroxides, for example ammonium peroxide, sodium peroxide, potassium peroxide, hydrogen peroxide, benzoyl peroxide and butyl peroxide, and the like; redox initiator systems, for example those comprising oxidizing agents, such as persulfates (in particular ammonium or alkali metal persulfates and the like), chlorates and bromates (including inorganic or organic chlorates and/or bromates), and reducing agents, such as sulfites and bisulfites (including inorganic and/or organic sulfites or bisulfites), oxalic acid and ascorbic acid, and also mixtures of two or more of these compounds.
The preferred initiators are water-soluble initiators. Preference is given in particular to sodium persulfate and azobisamidinopropane hydrochloride.
In an alternative form, the polymerization can be initiated by irradiation using ultraviolet light. The amount of initiator used is generally an amount sufficient to carry out the initiation of the polymerization. Preferably, the initiators are present in an amount ranging from 0.001 to approximately 10% by weight, with respect to the total weight of the monomers, and are preferably in an amount of less than 2% by weight, with respect to the total weight of the monomers, a preferred amount lying in the range from 0.05 to 1% by weight, with respect to the total weight of the monomers. The initiator is added to the polymerization mixture either continuously or portionwise.
When it is desired to obtain copolymers of high molecular weight, it is desirable to add initiator during the polymerization reaction. The addition can be gradual or portionwise. The polymerization is carried out under reaction conditions which are effective in polymerizing the monomers (c) and the monomers (a) in an atmosphere devoid of oxygen. Preferably, the reaction is carried out at a temperature ranging from approximately 300 to approximately 1000 and preferably between 600 and 90° C. The atmosphere devoid of oxygen is maintained throughout the duration of the reaction, for example by flushing with nitrogen throughout the reaction.
Use may be made in particular of “living” or “controlled” radical polymerization methods. These methods are particularly useful for the preparation of controlled structure copolymers.
Reference may in particular be made, as examples of “living” or “controlled” polymerization processes, to:
When grafted or comb controlled architecture copolymers are involved, the latter can be obtained by “direct grafting” and “copolymerization” methods.
Direct grafting consists in polymerizing the chosen monomer(s) by the radical route in the presence of the polymer selected to form the backbone of the final product. If the monomer/backbone pair and the operating conditions are carefully chosen, then there may be a transfer reaction between the growing macroradical and the backbone. This reaction generates a radical on the backbone and it is starting from this radical that the graft grows. The primary radical resulting from the initiator can also contribute to the transfer reactions.
As it relates to the copolymerization, it employs, in a first step, the grafting, at the end of the future pendant segment, of a functional group which can be polymerized by the radical route. This grafting can be carried out by conventional methods of organic chemistry. Then, in a second step, the macromonomer thus obtained is polymerized with the monomer chosen to form the backbone and a “comb” polymer is obtained. The grafting can advantageously be carried out in the presence of a polymerization control agent, such as mentioned in the above references.
The processes for the preparation of star-shaped polymers can essentially be classified into two groups. The first corresponds to the formation of the arms of the polymers starting from a multifunctional compound constituting the center (core-first technique) (Kennedy, J. P. et al., Macromolecules, 29, 8631 (1996), Deffieux, A. et al., ibid, 25, 6744, (1992), and Gnanou, Y. et al., ibid, 31, 6748 (1998)) and the second corresponds to a method where the polymer molecules which will constitute the arms are first synthesized and subsequently bonded together to a core to form a star-shaped polymer (arm-first technique).
Reference may be made, as an example of the synthesis of this type of polymer, to patent WO 00/02939. Mention may also be made of polymerization processes starting from a core comprising several transfer groups and of micelle crosslinking processes.
The polymer according to the invention can be used in particular as emulsifying or coemulsifying agent for preparing or stabilizing emulsions. It can, for example, be used in emulsions, one phase of which is a silicone oil. It can also be used to render compatible several compounds within a formulation. It can also be used as agent for helping with the deposition of another compound or as initiator of the deposition of another compound. It can be of use in carrying a compound, for example a silicone, to a surface.
The polymer can in particular be used in cosmetic compositions, in detergent compositions for the care of the home, in compositions for caring for the laundry, or as molecular recognition agent, or as transmembrane passage agent, or as additive for paper pulp, coating composition for paper, paint, for example paint for wood. Mention may be made, as cosmetic compositions, of shampoos, conditioners, shower gels or creams for caring for the skin. These compositions can additionally comprise at least one anionic and/or amphoteric surfactant and optionally agents such as silicone oils, nonsilicone oils or polysaccharides which are optionally modified. In these compositions, the polymer can contribute conditioning effects, effects of helping with the conditioning, sensory or “cosmetic” effects, effects of feel, of softness, of suppleness, of helping in disentangling, of gloss, of ability to be styled on dry or wet hair.
Other details or advantages of the invention will become apparent in the light of the examples below, which do not have a limiting nature.
Cellobiose (Fluka) (5 g, 14.6 mmol) is dissolved in allylamine (Aldrich) (150 ml).
The reaction mixture is kept stirred magnetically at ambient temperature for 72 h. Thin layer chromatography (“TLC”, ethyl acetate/petroleum ether 1/1) is carried out on an aliquot acetylated according to a conventional method (pyridine/acetic anhydride 1/1). After evaporating to dryness, the product obtained is a white powder.
The crude reaction product is selectively N-acetylated in a methanol/acetic anhydride solution (100 ml, 5/1, v/v). The conversion is monitored by thin layer chromatography (acetonitrile/water 7/3). The solution is left stirring for 4 h and then evaporated to dryness after addition of methanol (3 times). TLC shows the formation of a second compound which is probably O-acetylated. In order to remove it, the crude product is taken up in methanol (100 ml) and a 1M MeONa solution is added dropwise until a pH of 10 is obtained. This pH is determined by deposition of a drop of reaction mixture on a strip of moistened pH paper. Monitoring by TLC shows the disappearance of the O-acetylated compound. The solution is subsequently neutralized on Amberlite IR 120H+ resin, filtered, evaporated to dryness and lyophilized. Product 2 is obtained with a quantitative yield (6.18 g).
1H NMR (300 MHz, D2O, 353K)
δ=5.86-5.98 (dddd-oct, 1H, CH═CH2), 5.32 (d, 1H, J1.2=8.04 Hz, H1β), 5.32-5.15 (m, 2H, —CH═CH2), 4.54 (d, H1II), 4.05-3.91 (m, 2H, —CH2—CH═CH2), 3.83-3.77 (m, 2H, H-6II), 3.75-3.71 (m, H-6), 3.75-3.67 (m, 4H, H-2, H-3, H-4, H-5), 3.56-3.31 (m, 4H, H-3II, H-5II, H-4II, H-2II), 2.23 (s, 3H, —CH3 (Ac)).
13C NMR (75 MHz, D2O, 300K)
δ=178.41 (—C═O), 135.4 (CH═CH2), 117.76 (CH═CH2), 103.44 (C-1II), 83.42 (C-1), 79.45 (C-2), 77.88 (C-3), 77.07 (C-3II), 76.75 (C-5), 76.34 (C-5II), 74.30 (C-2II), 70.71 and 68.91 (C-4 and C-4II), 61.85 (C-6II), 61.41 (C-6), 42.49 (CH2—CH═CH2), 24.33 (—CH3 (Ac)).
MS (FAB+): m/z=424 [M+H]+
m/z=446 [M+Na]+.
Product 2 (5 g, 11.8 mmol), taken up in a minimum amount of water (25 ml) in a photochemical cell, is treated with cysteamine (2-aminoethanethiol hydrochloride, 98%, Acros Organics) (9.36 g, 82.6 mmol, 7 eq.).
The entire contents are irradiated (254 nm) under argon and kept stirred magnetically at ambient temperature for 24 h. Thin layer chromatography reveals the presence of product 3. The latter is purified on a column of ion-exchange resin (Dowex X 50 WX4) of H+ ionic form and is successively eluted with H2O and 0.1M NH4OH. Product 3 is subsequently lyophilized and is obtained with a yield of 65% (3.8 g, 7.68 mmol).
1H NMR (300 MHz, D2O, 353K)
δ=5.02 (d, 1H, J1.2=8.04 Hz, H1β), 4.56 (d, 1H, H1II), 4.01-3.36 (m, 16H), 2.75-2.63 (m, 4H, S—CH2 and CH2—S) 2.27 (s, 3H, CH3 (Ac)), 1.94 (m, 2H, —CH2)
13C NMR (75 MHz, D2O, 303K)
δ=176.6 and 175.74 (—C═O), 102.86 (C-1II), 87.22 (C-1), 78.47 and 77.39 (C-2 and C-3), 77.17 (C-3II) 76.39 (C-5II), 75.75 (C-5), 73.52 (C-2II), 70.31 (C-4), 69.84 (C-4II), 60.99 (C-6II), 60.46 (C-6), 40.65 (—CH2), 29.72 (—CH2), 29.50-28.62-28.23 (3*—CH2), 21.90-21.73 (—CH3 (Ac)).
MS (FAB+): m/z=501 [M+H]+
Product 3 (5 g, 10 mmol) is dissolved in a water/methanol mixture (75 ml; 1/1, v/v) in the presence of sodium carbonate (7.7 g). The medium is kept stirred magnetically at 0° C. while a solution of acryloyl chloride (4.6 ml, 56.9 mmol., Fluka) and THF (35 ml) is added gradually over 5 min. Thin layer chromatography (CH3CN/H2O: 6/4) shows complete conversion of product 3 to a compound having an Rf=0.6. The mixture is taken up in 300 ml of water, then reconcentrated and taken up once more in 200 ml of water in the presence of a radical inhibitor (2,6-di(tert-butyl)-4-methylphenol) (7.7 ml of a THF solution comprising 0.5% of inhibitor). Product 4 is concentrated, then purified on a column of C18 silica gel and lyophilized (5.5 g, 100%).
1H NMR (300 MHz, D2O, 353K)
δ=6.249 (m, 2H, CH═CH2), 5.799 (dd, 1H, CH═CH2), 4.95 (d, 1H, J1.2=7.68 Hz, H1β), 4.55 (d, 1H, H1II), 4.17-3.31 (m, 16H), 2.79 (m, 2H, NCH2CH2CH2S), 2.65 (m, 2H, NCH2CH2CH2S), 2.24 (s, 3H, CH3 (Ac)), 1.94 (m, 2H, NCH2CH2CH2S).
13C NMR (75 MHz, D2O, 303K)
δ=175.89 and 168.95 (—C═O) 130.35 (CH═CH2) and 127.80 (CH═CH2), 102.90 (C-1II), 87.27 (C-1), 78.57 and 77.46 (C-2 and C-3), 77.24 (C-3II), 76.41 (C-5II), 75.87 (C-5), 73.57 (C-2II), 70.33 (C-4), 69.87 (C-4II), 60.99 (C-6II), 60.58 (C-6), 39.16 (—CH2), 30.92, 30.70, 28.99, 28.37 (4*-CH2), 21.76 (—CH3 (Ac)).
MS (FAB+): m/z=577 [M+Na]+.
High resolution mass spectrum (ESI+): C22H38N2O12S
Value calculated: m/z=577.20432 [M+Na]+
Value measured: m/z=577.2043 [M+Na]+
Molar ratio: 95% MAPTAC, 5% product 4
Method: Introduction of MAPTAC and product 4 into a closed stirred reactor
Product 4 (0.376 g) and MAPTAC (6 g, Aldrich) are diluted in a minimum amount of water (3 g) at 80° C. under a stream of nitrogen. The V50 is injected every hour for three hours. The polymerization follows the following protocol:
After ultrafiltration over a 10 KDa membrane, the polymer is obtained with a yield by weight of 83%.
The number-average molar mass (Mn) and the weight-average molar mass (Mw) are measured by GPC coupled to MALS and conductimetry under the following conditions:
Mw: 1 244 000 g/mol
Mn: 360 000 g/mol
Stage 1: Hydrolysis of xyloglucans. Production of the XXXG, XXLG (or XLXG) and XLLG oligomers (products 5, 6 and 7) by cellulase 3042A
The operation is carried out on a mixture comprising the DPs 7, 8 and 9, respectively the XXXG, the XXLG (or XLXG) and the XLLG, in a molar ratio of 15%, 35% and 50%.
13 g of tamarind seed xyloglucan (3A, Dainippon Pharmaceutical) are suspended in distilled water (1 l) at 37° C. with stirring. After dissolution, cellulase 3042A (2.7 ml) is then added to the medium. The mixture is stirred for four hours. Thin layer chromatography (CH3CN/H2O: 7/3) shows the formation of products 5, 6 and 7.
The solution is subsequently brought to reflux, in order to denature the enzyme, filtered and lyophilized. Two successive ultrafiltrations are carried out with 500 Da and 10 000 Da membranes. After these ultrafiltrations, the mixture of products 5, 6 and 7 is obtained with a yield by weight of 80%.
MS (MALDI-TOF): (5) m/z=1085 [M+Na]+
(6) m/z=1247 [M+Na]+
(7) m/z=1409 [M+Na]+
The mixture of 5, 6 and 7 (5 g) is dissolved in allylamine (100 ml, Aldrich). The reaction mixture is kept stirred magnetically at ambient temperature for 4 days. After evaporating to dryness (coevaporation with toluene), the mixture obtained is a white solid which is selectively N-acetylated overnight in 1 l of a MeOH/Ac2O solution (20/1, v/v). The conversion is monitored by thin layer chromatography (CH3CN/H2O: 6/4). Products 8, 9 and 10 are subsequently concentrated and lyophilized (4.9 g, 94%)
MS (MALDI-TOF): (8) m/z=1166 [M+Na]+
(9) m/z=1328 [M+Na]+
(10) m/z=1490 [M+Na]+
1H NMR (300 MHz, D2O, 353K)
δ=5.96 (dddd, 1H, CH═CH2), 5.29 (m, 1H, CH═CH2), 5.18 and 4.98 (d, 1H, H1xyl), 4.58 (dd, 1H, H1gluc and gal), 4.10-3.41 (m, H), 2.27 (s, 3H, CH3 (Ac)).
The mixture of 8, 9 and 10 (20 g, 15 mmol), taken up just to solubility in the minimum amount of distilled water (250 ml), is treated with cysteamine (2-aminoethanethiol hydrochloride, 98%, Acros Organics) (8.69 g, 5 eq.).
The solution is irradiated at 254 nm in a quartz photochemical cell kept under argon. The reaction mixture is left under magnetic stirring for 48 h. Thin layer chromatography (CH3CN/H2O: 1/1) shows virtually complete conversion of a mixture. Products 11, 12 and 13 are washed with methanol to remove the excess cysteamine, filtered through a Büchner funnel and purified on an ion-exchange resin (Dowex 50WX4) of H+ ionic form activated by 0.5M HCl and eluted successively with H2O and 0.1M NH4OH. 11, 12 and 13 are lyophilized (17 g, 81%)
MS (MALDI-TOF): (11) m/z=1221 [M+H]+
(12) m/z=1383 [M+H]+
(13) m/z=1545 [M+H]+
1H NMR (300 MHz, D2O, 353K)
δ=5.19 and 4.98 (d, 1H, H1xyl), 4.61 (dd, H, H1gluc and gal) δ 4.06-3.41 (m, H), 2.80 (m, 2H, NCH2CH2CH2S), 2.68 (m, 2H, NCH2CH2CH2S), 2.28 (s, 3H, CH3 (Ac)), 1.96 (m, 2H, NCH2CH2CH2S).
Products 11, 12 and 13 (7.8 g, 5.43 mmol) are dissolved in a water/methanol mixture (40 ml; 1/1) in the presence of sodium carbonate (4 g). The medium is kept stirred magnetically at 0° C. while a solution of acryloyl chloride (2.4 ml, 29.6×10−3 mol, Fluka) and THF (20 ml) is gradually added over 5 min. The reaction is monitored by thin layer chromatography (CH3CN/H2O: 6/4). The mixture is taken up in 120 ml of water, then reconcentrated and taken up once more in 80 ml of water in the presence of a radical inhibitor (2,6-di(tert-butyl)-4-methylphenol) (100 μl of a 0.5% THF solution). The mixture of 14, 15 and 16 is concentrated, then purified on a column of C18 silica gel and lyophilized (8 g, 100%).
MS (MALDI-TOF): (14) m/z=1297 [M+Na]+
(15) m/z=1459 [M+Na]+
(16) m/z=1621 [M+Na]+
1H NMR (300 MHz, D2O, 353K)
δ=6.32-6.20 (m, 2H, CH═CH2), 5.80 (dd, 1H, CH═CH2), 5.18 and 4.98 (d, 2H, H1xyl), 4.61 (d, H, H1glc and gal) δ 4.09-3.40 (m, H), 2.84 (m, 2H, NCH2CH2CH2S), 2.68 (m, 2H, NCH2CH2CH2S), 2.26 (s, 3H, CH3 (Ac)), 1.95 (m, 2H, NCH2CH2CH2S).
Molar ratio: 95% MAPTAC, 5% mixture of 14, 15 and 16
Method: introduction of MAPTAC and the mixture into a closed stirred reactor
The products 14, 15 and 16 (0.976 g) and the MAPTAC (6 g, Aldrich) are diluted in the minimum amount of water (7 g) at 80° C. under a stream of nitrogen. The V50 is injected every hour for three hours. The polymerization follows the protocol:
250 μl samples are taken every hour for kinetic studies and hydroquinone (reaction inhibitor) is added to each sample.
After ultrafiltration over a 10 KDa membrane, the polymer is obtained with a yield by weight of 83%.
Mw: 3 047 000 g/mol
Mn: 795 000 g/mol
Cellobiose (Fluka) (5 g, 14.6 mmol) is dissolved in allylamine (Aldrich) (150 ml).
The reaction mixture is kept stirred magnetically at ambient temperature for 72 h. Thin layer chromatography (“TLC”, ethyl acetate/petroleum ether 1/1) is carried out on an aliquot acetylated according to a conventional method (pyridine/acetic anhydride 1/1). After evaporating to dryness, the product obtained is a white powder.
The crude reaction product is selectively N-acetylated in a methanol/acetic anhydride solution (100 ml, 5/1, v/v). The conversion is monitored by thin layer chromatography (acetonitrile/water 7/3). The solution is left stirring for 4 h and then evaporated to dryness after addition of methanol (3 times). TLC shows the formation of a second compound which is probably O-acetylated. In order to remove it, the crude product is taken up in methanol (100 ml) and a 1M MeONa solution is added dropwise until a pH of 10 is obtained. This pH is determined by deposition of a drop of reaction mixture on a strip of moistened pH paper. Monitoring by TLC shows the disappearance of the O-acetylated compound. The solution is subsequently neutralized on Amberlite IR 120H+ resin, filtered, evaporated to dryness and lyophilized. Product 2a is obtained with a quantitative yield (6.18 g).
1H NMR (300 MHz, D2O, 353K)
δ=5.86-5.98 (dddd-oct, 1H, CH═CH2), 5.32 (d, 1H, J1.2=8.04 Hz, H1β), 5.32-5.15 (m, 2H, —CH═CH2), 4.54 (d, H1II), 4.05-3.91 (m, 2H, —CH2—CH═CH2), 3.83-3.77 (m, 2H, H-6II), 3.75-3.71 (m, H-6), 3.75-3.67 (m, 4H, H-2, H-3, H-4, H-5), 3.56-3.31 (m, 4H, H-3II, H-5II, H-4II, H-2II), 2.23 (s, 3H, —CH3 (Ac)).
13C NMR (75 MHz, D2O, 300K)
δ=178.41 (—C═O), 135.4 (CH═CH2), 117.76 (CH═CH2), 103.44 (C-1II), 83.42 (C-1), 79.45 (C-2), 77.88 (C-3), 77.07 (C-3II), 76.75 (C-5), 76.34 (C-5II), 74.30 (C-2II), 70.71 and 68.91 (C-4 and C-4II), 61.85 (C-6II), 61.41 (C-6), 42.49 (CH2—CH═CH2), 24.33 (—CH3 (Ac)).
MS (FAB+): m/z=424 [M+H]+
m/z=446 [M+Na]+.
Cysteamine (2-aminoethanethiol hydrochloride, 98%, Acros Organics) (25 g, 0.22 mol, 3.7 eq.) and V-50 (α,α′-azodiisobutyramidine dihydrochloride, 98%, Fluka) (16 g, 59 mmol, 1 eq.) are added to a solution of product 2a (25 g, 59 mmol) in water (400 ml). The reaction mixture is stirred at 60° C. for 2 h under an argon atmosphere. The reaction is monitored by thin layer chromatography (AcOEt/AcOH/H2O 3/3/2 v/v/v). The solution is subsequently purified on a column of ion-exchange resin (Dowex X 50 WX4) of H+ ionic form and eluted successively with H2O and 0.05M and then 0.1M NH4OH. The product 3a is subsequently lyophilized and is obtained with a yield of 95% (28 g, 56 mmol).
Product 3a (5 g, 10 mmol) is dissolved in a water/methanol mixture (75 ml; 1/1, v/v) in the presence of sodium carbonate (7.7 g). The medium is kept stirred magnetically at 0° C. while a solution of acryloyl chloride (4.6 ml, 56.9 mmol, Fluka) and THF (35 ml) is added gradually over 5 min. Thin layer chromatography (CH3CN/H2O: 6/4) shows complete conversion of product 3a to a compound having an Rf=0.6. The mixture is taken up in 300 ml of water, then reconcentrated and taken up once more in 200 ml of water in the presence of a radical inhibitor (2,6-di(tert-butyl)-4-methylphenol) (7.7 ml of a THF solution comprising 0.5% of inhibitor). Product 4a is concentrated, then purified on a column of C18 silica gel and lyophilized (5.5 g, 100%).
1H NMR (300 MHz, D2O, 353K)
δ=6.249 (m, 2H, CH═CH2), 5.799 (dd, 1H, CH═CH2), 4.95 (d, 1H, J1.2=7.68 Hz, H1β), 4.55 (d, 1H, H1II), 4.17-3.31 (m, 16H), 2.79 (m, 2H, NCH2CH2CH2S), 2.65 (m, 2H, NCH2CH2CH2S), 2.24 (s, 3H, CH3 (Ac)), 1.94 (m, 2H, NCH2CH2CH2S).
13C NMR (75 MHz, D2O, 303K)
δ=175.89 and 168.95 (—C═O) 130.35 (CH═CH2) and 127.80 (CH═CH2), 102.90 (C-1II), 87.27 (C-1), 78.57 and 77.46 (C-2 and C-3), 77.24 (C-3II), 76.41 (C-5II), 75.87 (C-5), 73.57 (C-2II), 70.33 (C-4), 69.87 (C-4II), 60.99 (C-6II), 60.58 (C-6), 39.16 (—CH2), 30.92, 30.70, 28.99, 28.37 (4*-CH2), 21.76 (—CH3 (Ac)).
MS (FAB+): m/z=577 [M+Na]+.
High resolution mass spectrum (ESI+): C22H38N2O12S
Value calculated: m/z=577.20432 [M+Na]+
Value measured: m/z=577.2043 [M+Na]+
Number | Date | Country | Kind |
---|---|---|---|
0407818 | Jul 2004 | FR | national |
Filing Document | Filing Date | Country | Kind | 371c Date |
---|---|---|---|---|
PCT/FR05/01799 | 7/12/2005 | WO | 00 | 4/28/2008 |