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The eruption of 'mommy porn' typified in E.L James' 50 Shades series has been argued by some as a marker for female sexual empowerment. I will agree that it has enlightened a change in coffee table conversation, in a similar way to the emergence of Ann Summers' parties; but here's the rub - the series isn't actually representative of BDSM or female empowerment - it's simply about male possession.
One of the books that detainees at Guantánamo Bay are 50 Shades of Grey, the international bestseller from the US writer EL James. Could it be that the US military authorities have decided that James' erotic thriller is actually pornographic (so-called "mummy porn") and therefore unsuitable for the camp's 155 detainees?
Lauren Oliver is the best-selling author of Before I Fall and the Delirium trilogy. The final book in the latter series - Requiem - has just been released. It wraps up the story of Lena, a plucky young woman who lives in a future America where love is classified as an illness and young people are "cured" of the disease.
It was reported by The Telegraph recently that 'sex has been all but eradicated from Hollywood scripts over the past 18 months'. Apparently sex scenes are being shunned in favour of dazzling special effects and tellingly, only two films containing sex and nudity made the box office top ten in the UK last year. So what is the reason for Hollywood's languishing libido?
Despite the growth of online markets and digital applications the mark of success is still "getting off" the Internet and into "the real world" a gathering of publishers and tech start-ups has been told.
I had always defended the BDSM and kink communities against charges of unsound, destructive desire. Indeed, I'd already worked as a professional dominatrix - and one with stringent ethics.
In short: Fifty Shades of Grey is erotica and thus disqualified from the Bad Sex in Fiction Award 2012 (or indeed in any other year). Phew.
The rise of the ebook is crucial to the emerging success of Indie Authors. By cutting out the middlemen - agents, publishers, traditional bookshops - authors can realise better returns and connect more directly with their readers. Most notable, of course, is the phenomenon of E.L. James' Fifty Shades of Grey - now the best-selling book in British history.
Today, girls may know more about sex but Lace's message of empowerment and equality is as relevant and important as it was 30 years ago. It's a pity that modern novels, especially informative ones that involve women's sexuality are put down as 'mummy porn', 'bonkbusters', 'bodice rippers', 'beach-reads', 'w*nk-fodder' or, simply, trash. But what has clearly been proved, and what has changed in the last 30 years, is that women are far more openly interested in having an enjoyable sex life.
No one should be forced to perform sexual acts that they feel uncomfortable with. But to blame books for creating a new and dangerous generation of sexually demanding women is ludicrous and harks back to the days when novel-reading was considered an unsuitable activity for women.
'Keep failing and fail better' says Samuel Beckett, the absurdist playwright, which has been my mantra this week.
Fifty Shades is basically Twilight with added S&M. This is not just romance, this is S&M romance, to paraphrase an advertising campaign familiar to yummy mummies everywhere.
You pay your money, stand in a field, and a man in a vest with a wispy moustache shouts at you for an hour. While this happens, your body is forced into all kinds of unnatural positions known to acolytes of the craft as "the press-up", "the star jump", the "run to the fence and back MOVE!" and - I shudder at the memory - "the burpee".
For some time the boys had talked about the summer holidays with some excitement, mainly because they were so looking forward to going to see their nanny who now lives in Tenerife.
On the whole sex/ erotica point, it's worth noting how little actually appears. It takes over 100 pages before there's actually any sex, and on the few occasions that there is it's over within a couple of pages. When it does appear it can be reasonably explicit, but it's also clumsy and evasive enough not to feel too pornographic.
Someone sent me a link the other day asking me to help save a local bookshop, "Could you tweet this? He needs help." She said and I opened th...


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Current Time
Spain nightlife
Spanish nightlife is legendary, and for good reason. This is the country that brought the world Ibiza-style house music and the international nightclub group Pacha. Yet not every night has to pulse with the beats of electronic and house music. Sophisticated cocktail bars are more popular than ever in Spain, as are theme bars (ever hear of Spanish country music?). Bohemian bars like Barcelona's Tinta Roja, with poetry readings, comedy nights, and classic film screenings, abound in the artsy districts of the big cities.
City dwellers tend to hit the first bar of the night after dinner ends (around 1 am), and then migrate to thediscotecas, or nightclubs, by 3 am, dancing till dawn. Cover charges vary widely, but you should expect to pay upward of 20 euros in classy venues. Dress to impress; trendy nightspots have strict dress codes and only allow those who look the part to cross their thresholds.
The Spaniards' deeply ingrained love for fiestas means that any city or town can host a good party, but a few stand out. Madrid residents are known as gatos (cats) for their late-night prowling; the city is famous for 3 am traffic jams caused by throngs of partiers. Seaside resorts, especially on the islands of Ibiza, Mallorca, and Gran Canaria, are also known for their nightlife.
Toss back a few cañas at this convivial Casco Viejo bar, which stays hopping late into the evening. There's an excellent selection of cheeses, which you...more
see the Bilbao + Basque Country guideThis multi-story town house is a spectacular-looking pile framed by palm trees beside Palma's port. Staircases on the outside lead to six different bars and...more
see the Ibiza + Mallorca guideHidden away by the Puente Isabel II, better known as the Triana Bridge, this outdoor hot spot is Seville's answer to the beach bar, with shady palm trees and...more
see the Seville guideSuite, a gay hot spot with a mixed crowd not far from Puerta del Sol, is anything you want it to be. There's a trendy restaurant; there's a bar that teems...more
see the Madrid guideSpace arrived in Plaça Espanya from Ibiza in 2004, and since then has risen through the ranks to become one of Barcelona's most fashionable nightclubs....more
see the Barcelona guideThe legendary club Space celebrated its 20th anniversary in 2009 by completely refitting the dance area, adding new high-tech light shows and a very...more
see the Ibiza + Mallorca guideWhile one usually imagines left-wing intellectuals of old hanging out in cafés, the legendary nightclub Sala Bikini was where the Franco-opposing gauche...more
see the Barcelona guideRick's in Casablanca wasn't a gay barat least not openlybut its Madrid namesake is. The Humphrey Bogart theme is fleshed out with marble floors,...more
see the Madrid guideCurrently the hottest venue for big-name bands visiting Barcelona, Razz is everything superstadiums are not. Small-scale—even the most vertically...more
see the Barcelona guidePuro Beach is on a man-made island in the Bay of Palma and attached to the Puro Hotel in town, though it's open to discerning non-guests, too. From the...more
see the Ibiza + Mallorca guide

subset: commoncrawl


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Rennes goalkeeper Nicolas Douchez insists that speculation linking him with a move to Aston Villa is pure fabrication.
The 30-year-old shot-stopper has been the subject of reports suggesting he turned down a move to Villa during the January transfer window.
Douchez is out of contract in the summer but denies that he will move to the Premier League club, indicating there is a chance he will remain at Rennes.
He told France Football: "Aston Villa? It was pure fabrication, a real stupidity that forced Gerard Houllier to deny.
"Let's say there is a 20 per cent chance I could stay at Rennes.
"I will be a free agent next summer and I want to take time to have no regrets. I was sure to leave, now it is less sure."
Douchez moved to Rennes in 2008 having enjoyed previous spells with Le Havre, Chateauroux and Toulouse.


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We get a better idea of how powerful Diana is and her character...
- She breaks out of one of GL's constructs & punches him across town!
- She punches Superman & he mentions he is gonna feel that one for awhile!
- We also get to see she has a short temper!
- When Batman tells her the JL does not cut the heads off their enemies she tells Batman she does! (I love a Wonder Woman who will kill!)
- She still has STRONG feeling for Steve!
All in all a great issue with a strong focus on Diana & her relationship with Steve!


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Did I just hear that?
Yes he said it
Torches and pitchforks
At least clowns aren't quite as creepy as scarecrows.
Was it the Gus Johnson imposter? Go get him!
It's all snark from here on out
I'm emailing JP.
When Prepared, Fear No One.
Can we all sign a potion on change.org to get channel 13 shut down?
Category 5 Boat Shoes Campus Rep PM for Discounts / Promo Codes
Where is Murph?
Takes one to know one.
Yes, that ruins an otherwise perfect day for us
Yeah, I really don't give a ****...
slip of the toungue. relax people.
Originally Posted by CycloneRulzzz mtown is among the worst to ever play this game. Originally Posted by hawkeye nation If [Iowa] were a good shooting team we could make a run at the final 4.
Who said it? The new guy in KC? If so, not surprised he's from kc, so prob a ku or mu fan.
Originally Posted by mtowncyclone13 slip of the toungue. relax people. You mean they meant to talk about themselves?
Let my Fred's Posse Ride: Georges, Naz, Hogue, Bryce, Nader, Monte, Matt, and McKay.
Forum Rules


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'Pushing Daisies' Movie Kickstarter Needs Way More Money Than 'Veronica Mars'
by Kevin Fitzpatrick March 28, 2013 @ 5:00 PM
Though no TV series has officially won the race to be the next 'Veronica Mars,' reviving a dormant property with a fan-funded feature film, cancelled ABC dramedy 'Pushing Daisies' ranks among those leading the charge. Series creator and 'Hannibal' helmer Bryan Fuller has been vocal about the possibility of a 'Pushing Daisies' Kickstarter project, but will it prove too expensive to attempt? Find out what Bryan Fuller has to say inside!


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Previous Episode | Next Episode
Through A Lens Darkly
by Dave KochThis page has been viewed 8 times this month, and 554 times total.
- Daria Cartoon Episode Guide
- MTV Animation
- Distributed by: Music Television, Sony Pictures Entertainment
- Cartoon Characters: Daria Morgendorffer, Quinn Morgendorffer, Helen Morgendorffer, Jake Morgendorffer, Jane Lane, Trent Lane, Brittany Taylor, Kevin Thompson, Jodie Landon, Charles "Upchuck" Ruttheimer III, Andrea, Angela Li, Timothy O'Neill, Amy Barksdale.
- Originally Released in 1999.
- Season: 3
- Color
- Running Time: 22 minutes.
Cartoon Summary:
Helen suggests that Daria get contact lenses after a bad-driving lesson, which Daria eventually agrees upon. However, Daria finds the contact lenses extremely uncomfortable and doesn't want to wear them anymore. But, not wanting to go back to the glasses, she winds up spending a day wearing neither one and bumping into people. Eventually, Brittany gives her some helpful advice about being herself.
Submit Additional Information:
Do you have anything to add to this page? Have we made any mistakes... or do you ahve any additional information about Through A Lens Darkly? If so, we would love to hear from you. Please send us a quick note with your additions or corrections to this page, and we will make the corrections as soon as possible!
Report Errors / Submit Additional Information
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Cartoon Comments:
Below are comments about this cartoon submitted by people who have seen this show. Take a look and see what others have said about Through A Lens Darkly. We want to know what you think, too!! Feel free to share your own opinion- we do want to hear from you! You can add your own comments here.
Through a Lens Darkly
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This episode meant a lot to me because addressed the issue of appearance vs. merit. I tend to rely on my talent to win friends and ifluence people, therefore ignoring my appearance.
Looking good is a full time job--and I donlt wanna be Quinn, but... (read more)
Looking good is a full time job--and I donlt wanna be Quinn, but... (read more)
4 of 7 people found this comment useful


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Sponsored by: Brakes Plus
May 24, 2010 5:55 PM by Greg Boyce
Eastbound Interstate 70 closed west of Limon at mile marker 247 because of high winds for more than an hour Monday afternoon.
At least one semi-trailer truck has tipped over in the wind. The road was closed at about 1:45 p.m. Monday and then reopened before 3 p.m.
1 month ago


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US 7683024 B2 Abstract The invention concerns novel biodegradable polyaminoacid materials, useful in particular for vectoring active principle(s). The invention also concerns novel pharmaceutical, cosmetic dietetic or phytosanitary compositions based on said polyaminoacids. The invention aims at providing a novel polymer raw material, capable of being used for vectoring active principles and enabling optimal fulfillment of all specified requirements: biocompatibility, biodegradability, easy and inexpensive transformation into particles vectoring active principles, the particles being themselves capable of forming colloidal suspensions, of being easily associated with numerous active principles, and of releasing said active principles in vivo. Therefor, the present invention concerns first of all amphiphilic polyaminoacids comprising aspartic acid units and/or glutamic acid units, characterized in that at least part of said units carry grafts, including at least one alpha-tocopherol motif, for example (polyglutamate or polyaspartate grafted with alpha-tocopherol of synthetic or natural origin).
Claims(24) 1. A polyamino acid comprising aspartic units and/or glutamic units, characterized in that at least some of these units bear side chains comprising at least one α-tocopherol unit.
2. The polyamino acid as claimed in
in which:
R1 represents H, a linear C2 to C10 or branched C3 to C10 acyl group, or a pyroglutamate;
R2 represents H, a C2 to C10 linear or C3 to C10 branched alkyl, benzyl or a terminal amino acid unit;
R3 is H or a cationic species selected from the group comprising of:
metallic cations selected from the subgroup consisting of sodium, potassium, calcium and magnesium,
organic cations selected from the subgroup consisting of:
amine-based cations,
oligoamine-based cations,
cations based on polyamine,
cations based on amino acid(s) selected from the class comprising cations based on lysine or arginine,
and cationic polyamino acids selected from the subgroup consisting of polylysine and oligolysine;
R4 represents a direct bond or a "spacer" based on 1 to 4 amino acid units;
A independently represented a —CH2— (aspartic unit) or —CH2—CH2— (glutamic unit) radical;
n/(n+m) ranges from 0.5 to 100 mol %;
n+m ranges from 3 to 1000;
T represents an α-tocopherol unit.
3. The polyamino acid as claimed in
4. The polyamino acid as claimed in
5. The polyamino acid as claimed in
6. The polyamino acid as claimed in
7. The polyamino acid as claimed in
8. The polyamino acid as claimed in
9. The polyamino acid as claimed in
10. The polyamino acid as claimed in
11. The polyamino acid as claimed in
12. The polyamino acid as claimed in
in which:
R′4 represents a direct bond or a "spacer" based on 1 to 4 amino acid units;
X is a hetero atom chosen from the group comprising oxygen, nitrogen and sulfur;
R5 and R6 independently represent H or a linear C1 to C4 alkyl;
n ranges from 3 to 1000.
13. The polyamino acid is claimed in
14. The polyamino acid as claimed in
15. A composition comprising at least one of the polyamino acids as claimed in any one of
16. The composition as claimed in
17. The composition as claimed in
18. The composition as claimed in
19. The composition as claimed in
20. The composition is claimed in
21. The composition as claimed in
22. The composition as claimed in
23. The composition as claimed in
wherein said medicinal product is formulated for oral, nasal, vaginal, ocular, subcutaneous, intravenous, intramuscular, intradermal, intraperitoneal or intracerebral administration,
wherein said active principle is selected from the group consisting of proteins, glycoproteins, proteins linked to one or more polyalkylene glycol chains, peptides, polysaccharides, liposaccharides, oligonucleotides, small organic molecules that are hydrophobic, small organic molecules that are hydrophilic and small organic molecules that are amphiphilic.
24. The polyamino acid of
Description This application is a 371 of PCT/FR03/50003, filed Jun. 3, 2003, which claims priority to FR 0207008, filed Jun. 7, 2002. The present invention relates to novel materials based on biodegradable polyamino acids, which are useful especially for the vectorization of active principal(s) (AP). The invention is also directed toward novel pharmaceutical, cosmetic, dietetic or plant-protection compositions based on these polyamino acids. These compositions may be of the type allowing the vectorization of AP and preferably being in the form of emulsions, micelles, particles, gels, implants or films. The APs under consideration are advantageously biologically active compounds that may be administered to an animal or human body via the oral, parenteral, nasal, vaginal, ocular, subcutaneous, intravenous, intramuscular, intradermal, intraperitoneal, intracerebral, buccal, etc. route. The APs more particularly concerned by the invention, but not limiting the invention thereto, are proteins, glycoproteins, peptides, polysaccharides, lipopolysaccharides, oligonucleotides, polynucleotides and organic molecules. However, they may also be cosmetic products or plant-protection products, such as herbicides, insecticides, fungicides, etc. In the field of vectorization of active principles, especially medicinal active principles, there is a need, in many cases:
To these ends, several types of polymers have been studied and some are even commercially available. Mention may be made, for example, of polymers of the polylactic, polylactic-glycolic, polyoxyethyleneoxypropylene, polyamino acid or polysaccharide type. These polymers constitute starting materials for manufacturing, for example, bulk implants, microparticles, nanoparticles, vesicles, micelles or gels. Besides the fact that these polymers must be suitable for manufacturing such systems, they must also be biocompatible, nontoxic, nonimmunogenic and cost-effective, and they should be easy to eliminate from the body and/or biodegradable. As regards this last aspect, it is furthermore essential that the biodegradation in the body should generate nontoxic products. By way of illustration of the prior art concerning polymers used as starting materials for making AP vectorization systems, various patents, patent applications or scientific articles are mentioned below. U.S. Pat. No. 4,652,441 describes polylactide microcapsules encapsulating the hormone LH-RH. These microcapsules are produced by preparing a water-in-oil-in-water emulsion comprising an aqueous inner layer containing the hormone, a substance (gelatin) for fixing said hormone, a polylactide oily layer, and also an aqueous outer layer (polyvinyl alcohol). The AP is released over a period of more than 2 weeks after subcutaneous injection. U.S. Pat. No. 6,153,193 describes compositions based on amphiphilic poly(oxyethylene)-poly(oxypropylene) micelles, for the vectorization of anticancer agents such as adriamycin. Akiyoshi et al. (J. Controlled Release 1998, 54, 313-320) describe pullulans that have been made hydrophobic by grafting cholesterol, and which form nanoparticles in water. These nanoparticles, which are capable of reversibility complexing with insulin, form stable colloidal suspensions. U.S. Pat. No. 4,351,337 describes amphiphilic copolyamino acids based on leucine and glutamate, which may be used in the form of implants or microparticles for the controlled release of active principles. Said active principles are released over a very long period that depends on the rate of degradation of the polymer. U.S. Pat. No. 4,888,398 describes polymers based on polyglutamate or polyaspartate, and optionally polyleucine, with pendent groups of alkyloxycarbonylmethyl type, placed randomly on the polyamino acid chain. These polyamino acids, grafted with side groups, e.g. methoxycarbonylmethyl, may be used in the form of biodegradable implants containing a sustained-release AP. U.S. Pat. No. 5,904,936 describes nanoparticles obtained from a polyleucine-polyglutamate block polymer, which are capable of forming stable colloidal suspensions capable of spontaneously associating with biologically active proteins without denaturing them. Said proteins may then be released in vivo in a controlled manner, over a long period. Patent application WO 00/30618 describes nanoparticles obtained from a poly(sodium glutamate) (polymethyl, ethyl, hexadecyl or dodecyl glutamate) block polymer, which are capable of forming stable colloidal suspensions capable of spontaneously associating with biologically active proteins without denaturing them. Said proteins may then be released in vivo in a controlled manner, over a long period. These amphiphilic copolyamino acids are modified by the presence of a hydrophobic alkyl side chain. U.S. Pat. No. 5,449,513 describes amphiphilic block copolymers comprising a polyoxyethylene block and a polyamino acid block, for example poly(β-benzyl-L-aspartate). These polyoxyethylene-polybenzylaspartate polymers form micelles that are capable of encapsulating hydrophobic active molecules such as adriamycin or indomethacin. Patent application WO 99/61512 describes polylysines and polyornithines functionalized with a hydrophobic group (palmitic acid linked to polylysine or ornithine and a hydrophilic group (polyoxyethylene). These polymers, for example polylysine grafted with polyoxyethylene and palmitoyl chains, form, in the presence of cholesterol, vesicles capable of encapsulating doxorubicin or DNA. It is moreover known practice to employ vitamin E derivatives, and more specifically α-tocopherol, to construct AP vectorization systems. Natural vitamin E consists of a mixture of compounds known as tocopherols (see Burton and Ingold, Acc. Chem. Res. 1986, 19, 194-201) and, in this mixture, the α-tocopherol derivative is largely in majority amount. Vitamin E and some of its derivatives are nowadays used as a source of vitamin or as antioxidant in foods and cosmetic products. For these common uses, the vitamin E is found in its D-α-tocopherol, form (its natural form) or in its D-L-α-tocopherol form (racemic and synthetic form). These two products are considered as being essentially nontoxic at doses considerably higher than therapeutic doses. The structure of α-tocopherol is as follows:
The chiral positions are marked with an asterisk. The natural form has the configurations R,R,R and the synthetic form is a mixture in which the chiral carbons are independently R or S. As regards the vitamin E derivatives used in the field of vectorization of active principles, there is, at the present time and to the inventors' knowledge, no α-tocopherol-based polymer product, with the exception of polymers of polyoxyethylene type, one end of which is grafted with α-tocopherol-succinate groups. Polyethylene glycol grafted with α-tocopherol succinate at the end of the chain (PEGylated vitamin E) is moreover available on the market, sold under the name TPGS 1000 by the company Eastman Chemical Ltd). This product, patented in 1954 (U.S. Pat. No. 2,680,749), is nowadays used as a source of oral-route vitamin E. This polymer, and similarly α-tocopherol-succinate and unmodified α-tocopherol, have been proposed for the vectorization of active principles. U.S. Pat. No. 5,869,703 describes similar compounds in which the polyoxyethylene chain comprise α-tocopherol at one end and a (meth)acrylic residue at its other end. These α-tocopherol derivatives are used to prepare stable amphiphilic vesicles (liposomes), used directly in cosmetic applications. Patent application WO 00/71163 describes formulations based on polyethylene glycol grafted with α-tocopherol-succinate at the end of the chain (TPGS 1000) and on α-tocopherol, for the solubilization of paclitaxel (anticancer product). At the present time, the toxicity associated with the polyoxyethylene part is not known, and it is known that polyoxyethylene is not degraded in vivo. Furthermore, this compound contains only one α-tocopherol unit per polymer chain and it has properties in solution similar to those of surfactants. In any case, the use of this product for vectorization would lead to sparingly stable polymer-active principle combinations. Patent EP 0 243 446 describes the use of α-tocopherol (hemi) succinate (organic acid derivative of α-tocopherol) for the manufacture of vesicles, in combination with an amine salt. These vesicles may be used for the encapsulation of various active principles including small molecules, peptides and proteins. In general, it is indicated in said patent that the organic acid may be an amino acid or a polyamino acid. However, no details are given in this respect. Only α-tocopherol (hemi) succinates are illustrated. Thus, even though there are a great many technical solutions in the prior art, developed and proposed for the vectorization of medicinal active principles, meeting all the requirements is difficult to achieve, and remains unsatisfactory. In this context, one of the essential objectives of the present invention is to provide a novel polymeric starting material, which may be used for AP vectorization and which can optimally satisfy all the specification details:
This objective, among others, is achieved by the present invention, which relates firstly to amphiphilic polyamino acids comprising aspartic units and/or glutamic units, characterized in that at least some of these units bear grafts comprising at least one α-tocopherol unit. These novel polymers have a biodegradable skeleton based on polaymino acids bearing side chains comprising α-tocopherol. These polymers have association and/or encapsulation properties that are surprising in comparison with similar products and, what is more, they are readily degraded in the presence of enzymes. The Applicant has, to its merit, had the idea of combining, in an entirely judicious and advantageous manner, particular biodegradable polyAsp and/or polyGul polyamino acids with grafts based on α-tocopherol (vitamin E), for the vectorization of AP. For the purposes of the invention, the term "polyamino acid" covers not only oligoamino acids comprising from 2 to 20 amino acid units, but also polyamino acids comprising more than 20 amino acid units. Preferably, the polyamino acids according to the present invention are oligomers or homopolymers comprising glutamic or aspartic amino acid repeating units or copolymers comprising a mixture of these two types of amino acid units, said units being partially substituted with grafts comprising α-tocopherol. The units under consideration in these polymers are amino acids having the D, L or D,L configuration and are linked via their α- or γ-positions for the glutamate or glutamic unit and the α- or β-position for the aspartic or aspartate unit. The preferred amino acid units are those having the L configuration and a bond of ai type. Even more preferably, the polyamino acids according to the invention correspond to the general formula (I) below: R1 represents H, a linear C2 to C10 or branched C3 to C10 acyl group, or a pyroglutamate; R2 represents H, a C2 to C10 linear or C3 to C10 branched alkyl, benzyl or a terminal amino acid unit; R3 is H or a cationic species preferably selected from the group comprising:
amine-based cations, oligoamine-based cations, cations based on polyamine (polyethyleneimine being particularly preferred), cations based on amino acid(s) advantageously chosen from the class comprising cations based on lysine or arginine,
R4 represents a direct bond or a "spacer" based on 1 to 4 amino acid units; A independently represented a —CH2— (aspartic unit) or —CH2—CH2— (glutamic unit) radical; n/(n+m) is defined as the molar degree of grafting and ranges from 0.5 to 100 mol %; n+m ranges from 3 to 1000 and preferably between 30 and 300; T represents an α-tocopherol unit. For these common uses, vitamin E is found in its D-α-tocopherol form (its natural form) or in its D,L-α-tocopherol form (racemic and synthetic form). These two products are considered as being essentially nontoxic at doses considerably higher than therapeutic doses. In the context of the invention, these two forms of α-tocopherol are preferred. The α-tocopherol is of natural or synthetic origin. According to a first embodiment of the invention, the polyamino acids are α-L-glutamate or α-glutamic homopolymers. According to a second embodiment of the invention, the polyamino acids are α-L-aspartate or α-L-aspartic homopolymers. According to a third embodiment of the invention, the polyamino acids are α-L-aspartate α-L-glutamate or α-L-aspartic/α-L-glutamic copolymers. Advantageously, the distribution of the aspartic and/or glutamic units bearing grafts comprising at least one α-tocopherol unit is such that the polymers thus composed are either random, or block type or of multi-block type. According to another mode of definition, the polyamino acids according to the invention have a molar mass of between 2000 and 100 000 g/mol and preferably between 5000 and 40 000 g/mol. It is moreover preferable for the molar degree of grafting with α-tocopherol of the polyamino acids according to the invention to be between 3% and 70% and preferably between 5% and 50%. The polyamino acids of the invention are, remarkably, capable of being used in several ways depending on the degree of grafting. The methods for forming a polymer for the encapsulation of an active principle in the various forms targeted by the invention are known to those skilled in the art. For further details, reference may be made, for example, to these few particularly pertinent references:
The polyamino acids are also extremely advantageous as a result of the fact that, at a relatively low degree of grafting of about from 3% to 10%, they form in water at pH 7.4 (for example with a phosphate buffer) colloidal suspensions or gels depending on the polymer concentration. Furthermore, the polyamino acid particles forming the dispersed phase of the colloidal suspension can readily associate with active principles such as proteins, peptides or small molecules. The preferred forming operation is that described in patent application WO 00/30618 by the Applicant, which consists in dispersing the polymer in water and incubating the solution in the presence of an AP. This solution can then be filtered through a 0.02 μm filter and then injected directly into a patient. Above a degree of grafting of 10%, the polymer can form microparticles capable of associating or of encapsulating APs. In this context, the forming of the microparticles may take place by codissolving the AP and the polymer in a suitable organic solvent, and the mixture is then precipitated from water. The particles are then recovered by filtration and can then be used for an oral administration (in the form of a gel capsule, in compacted and/or coated form, or alternatively in a form dispensed in an oil) or parenterally after redispersing in water. At degrees of grafting of greater than 30%, the redispersion of the polymer in aqueous phase becomes more difficult due to the smaller amount of ionizable carboxylate functions, and the polymer precipitates. In this case, the polymer can be dissolved in a biocompatible solvent such as N-methylpyrrolidone or a suitable oil such as Miglyol®, and then injected intramuscularly or subcutaneously or into a tumor. The diffusion of the solvent or of the oil results in precipitation of the polymer at the site of injection and thus forms a deposit. These deposits then provide controlled release by diffusion and/or erosion and/or hydrolytic or enzymatic degradation of the polymer. In general, the polymers of the invention, in neutral or ionized form, may be used alone or in a liquid, solid or gel composition and in an aqueous or organic medium. It should be understood that the polymer based on polyamino acids contains carboxylic functions that are either neutral (COOH form) or ionized, depending on the pH and composition. For this reason, the solubility in an aqueous phase depends directly on the content of free COOH (not grafted with vitamin E) and on the pH. In aqueous solution, the counter-cation may be a metallic cation such as sodium, calcium or magnesium or an organic cation such as triethanolamine, tris (hydroxymethyl) aminomethane or a polyamine such as polyethyleneimine. The polymers of the invention are obtained via methods known to those skilled in the art. The polyamino acids may be obtained in at least two ways:
In the first case, a polyamino acid, homopolyglutamate, homopolyaspartate or a glutamate/aspartate copolymer, in block, multiblock or random form, is prepared, for example, according to standard methods. To obtain polyamino acids of α type, the technique most commonly used is based on the polymerization of N-carboxyamino acid anhydrides (NCA), described, for example, in the article "Biopolymers, 1976, 15, 1869" and in the book by H. R. Kricheldorf "alpha-Aminoacid-N-carboxy Anhydride and related Heterocycles", Springer Verlag (1987). The NCA derivatives are preferably NCA-O—Me, NCA-O—Et or NCA-O—Bz derivatives (Me=methyl, Et=ethyl and Bz=benzyl). The polymers are then hydrolyzed under suitable conditions to give the polymer in its acid form. These methods are inspired from the description given in patent FR 2 801 226 from the Applicant. A certain number of polymers that may be used according to the invention, for example, of poly(α-L-aspartic), poly (α-L-glutamic), poly (α-D-glutamic) and poly (γ-L-glutamic) type of variable masses are commercially available. The polyaspartic of the α-β type is obtained by condensing aspartic acid (to obtain a poly-succinimide), followed by a basic hydrolysis (see Tomida et al. Polymer 1997, 38, 4733-36). The coupling of α-tocopherol with an acid function is readily performed by reacting the polyamino acid with vitamin E in the presence of a carbodiimide as coupling agent and, preferably, a catalyst such as 4-dimethyl-aminopyridine, and in a suitable solvent such as dimethylformamide (DMF), N-methylpyrrolidone (NMP) or dimethyl sulfoxide (DMSO). The carbodiimide is, for example, dicyclohexylcarbodiimide or diisopropylcarbodiimide. The degree of grafting is controlled chemically via the stiochiometry of the constituents and reagents, or the reaction time. In the second case, an NCA derivative of α-tocopherol having the structure below is synthesized. The synthesis is analogous to that described for stearyl glutamate N-carboxyanhydride by Poché et al. Macromolecules 1995, 28, 6745-53.
The NCA derivative of α-tocopherol-glutamate is then copolymerized with, for example, benzylglutamate NCA and, to obtain the glutamate or glutamic functions, a selective hydrolysis reaction of the benzyl functions is performed in a mixture of trifluoroacetic acid and hydrobromic acid at room temperature. It should be noted that this second synthetic route makes it possible easily to prepare random, block or multiblock copolymers simply by modifying the order of addition of the monomers. The coupling of vitamin E via a spacer consisting of 1 to 4 amino acids may be performed via successive reactions of vitamin E with suitably protected amino acids, which are then deprotected to have a graftable amine function on the polymer, or by reaction with an oligopeptide. For example, the synthesis of an α-tocopherol with a leucine unit is performed according to a general method well known to those skilled in the art and according to the following scheme:
It should be noted that the direct grafting of α-tocopherol onto the polymer takes place via an ester function, whereas in the case of the presence of a spacer based on amino acid(s), it takes place via an amide function. As for the preparation of an ester bond, the amide bond may be formed in the same manner using a standard coupling agent such as a dialkylcarbodiimide. According to one variant of the invention, the polyamino acids with which it is concerned not only bear α-tocopherol grafts but also, per molecule, at least one graft of polyalkylene glycol type linked to a glutamate and/or aspartate unit, and preferably of formula (II) below:
Preferably, the polyalkylene glycol is a polyethylene glycol. According to another preferred characteristic of the invention, the molar percentage of grafting of the polyalkylene glycol ranges from 1% to 30%. The grafting of these pendent side groups (II) is performed in a manner that is known per se and according to techniques within the capability of a person skilled in the art, for example by forming amide, ester or thioester bonds with the carboxyls of glutamate and/or aspartate monomers. These techniques may especially be those used for the grafting of α-tocopherol onto a polyamino acid skeleton, said techniques being described in the present patent application. According to another of its aspects, the invention is directed toward a pharmaceutical, cosmetic, dietetic or plant-protection composition comprising at least one of the polyamino acids as defined above. According to one advantageous arrangement of the invention, this composition comprises, besides α-tocopherol, at least one active principle, which may be a therapeutic, cosmetic, dietetic or plant-protection active principle. Preferably, the active principle is a protein, a glycoprotein, a polysaccharide, a liposaccharide, an oligonucleotide, a polynucleotide or a peptide. Even more preferably, the active principle is a hydrophobic, hydrophilic or amphiphilic organic "small molecule". According to the present description, the term "small molecule" especially denotes nonprotein molecules. This composition may be in the form of nanoparticles, microparticles, solutions, emulsions, suspensions, gels, micelles, implants, powders or films. According to one of its particularly preferred forms, the composition, containing or not containing active principle(s), is a stable colloidal suspension of nanoparticles and/or microparticles and/or micelles of polyamino acids, in an aqueous phase. The composition according to the invention, when it is pharmaceutical, may be administered via the oral, parenteral, nasal, vaginal, ocular, subcutaneous, intravenous, intramuscular, intradermal, interperitoneal, intracerebral or buccal route. It may also be envisioned for the composition to be in the form of a solution in a biocompatible solvent, capable of being injected subcutaneously, intramuscularly or into a tumor. According to another variant, the composition according to the invention is formulated such that it is injectable and such that it is capable of forming a deposit at the site of injection. The invention is also directed toward compositions comprising polyamino acids according to the invention and active principles, and which are capable of being used for the preparation:
According to yet another of its aspects, the invention is directed toward a process for preparing:
As indicated above, the techniques for associating one or more APs with the α-tocopherol-grafted polyamino acids according to the invention are described especially in patent application WO 00/30618. The invention also relates to a therapeutic treatment method that consists essentially in administering the composition as described in the present description, via the oral, parenteral, nasal, vaginal, ocular, subcutaneous, intravenous, intramuscular, intradermal, intraperitoneal, intracerebral or buccal route. According to one particular embodiment, the therapeutic treatment method consists essentially in using a composition as described above in the form of a solution in a biocompatible solvent, and then injecting it subcutaneously, intramuscularly or into a tumor, preferably such that is forms a deposit at the site of injection. As examples of APs that may be associated with the polyamino acids according to the invention, whether or not they are in the form of (nano or micro) particles, mention may be made of:
The invention will be understood more clearly and its advantages and implementation variants will emerge more clearly from the examples that follow, which describe the synthesis of α-tocopherol-grafted polyamino acids, their conversion into an AP vectorization system (stable aqueous suspension of nanoparticles) and demonstration of the capacity of such a system to associate with APs (small organic molecules, proteins, etc.) to form pharmaceutical compositions. Synthesis of a Polyglutamate Grafted with α-tocopherol of Synthetic Origin The α-L-polyglutamate polymer, with a mass equivalent to about 10 000 relative to a polyoxyethylene standard, is obtained by polymerization of NCAGLuOMe, followed by a hydrolysis, as described in patent application FR 2 801 226. 5.5 g of this α-L-polyglutamate polymer are dissolved in 92 ml of dimethylformamide (DMF), by heating at 40° C. for 2 hours. Once the polymer has dissolved, the temperature is allowed to return to 25° C. and 1.49 g of D,L,α-tocopherol (>98%, obtained from Fluka®) predissolved in 6 ml of DFM, 0.09 g of 4-dimethylaminopyridine predissolved in 6 ml of DMF and 0.57 g of diisopropylcarbodiimide predissolved in 6 ml of DMF are successively added. After stirring for 8 hours at 25° C., the reaction medium is poured into 800 ml of water containing 15% sodium chloride and hydrochloric acid (pH 2). The precipitated polymer is then recovered by filtration, washed with 0.1N hydrochloric acid and then washed with water. The polymer is then redissolved in 75 ml of DMF and then reprecipitated from water containing, as previously, salt and acid at pH2. After washing twice with water, the polymer is washed several times with diisopropyl ether. The polymer is then oven-dried under vacuum at 40° C. A yield of about 85% is obtained. The degree of grafting estimated by proton NMR is about 7.8% and an HPLC analysis reveals a residual tocopherol content of less than 0.3%. Mw (measured by GPC, eluting with NMP)=17 500 g/mol (as polymethyl methacrylate equivalent). Polymers containing variable amounts of tocopherol are prepared in the same manner.
In all cases, the amount of tocopherol effectively grafted was confirmed by NMR. Synthesis of a Polyglutamate Grafted with α-tocopherol of Natural Origin In a similar manner, the polymer P6 is synthesized with 7.3% of D-alpha-tocopherol of natural origin (at a purity of 98.5%, obtained from the company ADM France). The molar mass is 17 400 (GPC NMP, PMMA eq). The polymers are dissolved in a phosphate-buffered saline at pH 7.4 at concentrations ranging from 10 to 40 mg/ml, and the pH is adjusted to 7.4 by adding 0.1N sodium hydroxide. The dissolution is observed visually.
Phosphate buffer: 0.01M phosphate, 0.0027M KCl and 0.137M NaCl. An observation by electron transmission of the clear solutions of the polymer P1 deposited on a support shows the existence of nanoparticles of 15 to 25 nm. A comparative analysis of the solutions of polymer P1, P6 and α-tocopherol succinate at 15 mg/ml in water at pH 7.4 (phosphate buffer) reveals that only the α-tocopherol succinate develops a milky solution characteristic of vesicles, as described in patent EP 0 243 446. According to one of the subjects of the invention, the polymers may be used in the form of a colloidal suspension in water and associated with an active principle. For this application, it is demonstrated in the experiment below that with certain polymers, especially those with a degree of grafting of about from 5% to 10% tocopherol, the adsorption capacity is greater than that of a similar compound of the prior art. For this study, the polymer P1 was compared with a similar polymer containing a dodecanol chain grafted onto a polyglutamate. This polymer is described in patent WO 00/30618. The study is performed in the following manner: the polymers are dissolved in an aqueous solution at pH 7 (phosphate buffer) and 5 mg of the dye known as Orange OT (Rn CAS: 2646-17-5) are added. The solutions are left in an ultrasonic bath for 1 hour to achieve the association. The solutions are then centrifuged to remove the nonassociated dye, and the optical density is measured at the λmax of the dye, which is at 495 nm.
It is found that at a molar degree of grafting of less than a half and at a slightly smaller mass concentration of polymer, the polymer P1 has a much higher capacity for association of the dye Orange OT. Synthesis of a Polyglutamate Containing an α-tocopherol Leucine Graft The α-tocopherol leucine derivative is first synthesized, in the following manner. D,L-α-Tocopherol (4.3 g) is reacted with BOC-leucine (2.3 g) in 15 ml of dichloromethane in the presence of 4-dimethylaminopyridine (244 mg) and diisopropylcarbodiimide (1.5 g). After 2 hours at 30° C. the product is purified by filtration on a column of silica. 5 g of the product α-tocopherol leucine BOC are obtained (77% yield). Its structure is confirmed by NMR spectroscopy. The deprotection of the product is performed in trifluoroacetic acid at a temperature of between 5 and 10° C. for 1 hour. After purification by filtration through silica, 3.3 g of the desired product are isolated (78% yield). Its structure is confirmed by NMR spectroscopy. The grafting reaction on a polyglutamic acid is then performed under the same conditions as in example 1, with a degree of grafting of 7%. The structure of the polymer and the degree of grafting were confirmed by NMR spectroscopy. Synthesis of a polyglutamate containing an α-tocopherol graft and a polyoxyethylene glycol graft. A grafting reaction is performed as in example 1, with 11 mol % of α-tocopherol and 2 mol % of an amino methoxypolyethylene glycol of formula MeO(CH2CH2O)aCH2CH2NH2 and of molar mass 3000 (product obtained from the company Shearwaters). The polymer in its acid form is obtained in a yield of 72%. Proton NMR confirms a degree of grafting with α-tocopherol of 10.9% and with polyethylene glycol of 1.9%. A solution containing 1 mg of polymer P1 and 7 mg of insulin at pH 7.0 in 1 ml of water is prepared and is left to incubate for 2 hours. The suspension is then ultrafiltered (10 000×G, 20 minutes with a 100 KDa threshold). The free insulin in the filtrate is assayed by HPLC and the amount of associated insulin is deduced by difference. A degree of association of greater than 95% relative to the insulin employed is measured. Under the same conditions, the comparative polymer of example 8 allows 40% association. The adsorption capacity of polymer P1 is thus greater. Polymer P1 is dissolved at pH 7.5 (phosphate buffer and 10 mM of calcium cation) and at a concentration of 20 mg/ml. 0.1 ml of protease (solution of 10 mg/ml) is added and the degradation is monitored by aqueous GPC. A relatively rapid degradation is found, with a half-life time of the initial polymer of about 100 minutes. Patent Citations
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MoreExtending for 55 miles from Portsmouth Island to Shackleford Banks, Cape Lookout National Seashore includes 28,400 acres of uninhabited land and marsh. The remote, sandy islands are linked to the mainland by private ferries. Loggerhead sea turtles, which have been placed on the federal list of threatened and endangered species, nest here. To the south, wild ponies roam Shackleford Banks.
Four-wheel-drive vehicles are allowed on the beach, and primitive camping is allowed. There are primitive cabins (with and without electricity, no linens or utensils) with bunk beds. Private ferry service is available from Beaufort and Morehead City to Shackleford Banks, Beaufort, and Harkers Island to the Cape Lookout Lighthouse area, from Davis to Shingle Point, from Atlantic to North Core Banks and Long Point, and from Ocracoke Village to Portsmouth Village.