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I joined the circle, sitting cross-legged on the floor in the library conference room, and braced myself. Things were about to get weird.
Immediately, I sensed that Laughter Yoga draws two types of people: Those in desperate need of a laugh, and those who can't stop laughing. From outward appearance only, the group also covered both ends of the economic and age spectrums.
Not that I was judging, because the instructor explained that Laughter Yoga is a judgment-free place where participants are encouraged to engage in childlike play. She taught us to clap and say, "Very good, very good, YAY!"
The concept of gathering to laugh for better health originated in India in 1995, when Dr. Maden Kataria met with five people in Mumbai Park to test the benefits of laughter therapy. After the first few gatherings, Dr. Kataria ran out of jokes. He implemented laughter for no reason when he realized that the body can't differentiate between real and fake laughter.
In the years since, Laughter Yoga has spread to 70 countries, and research has proved the power of laughing. For starters, sustained laughter reduces stress hormone levels that elevate blood pressure, aids digestion, acts as a pain killer, eases depression, stimulates the heart, enhances circulation and strengthens the immune system. Laughing in a group also helps us feel connected with others.
The instructor led us through yogic breathing and asked us to laugh from deep within our diaphragms. Ho ho ho, ha ha ha. We laughed to the tune of Beethoven. We introduced ourselves and laughed. We looked like morons doing the conga line. It felt ridiculous miming throwing my problems onto the floor and then stomping on them, laughing. It also looked and felt seriously funny. My body was falling for the tricks. I felt like I was in a comedy club, two drinks in.
The hysterics hit when we were lying on our backs, waving our legs in the air like babies. It became a silent, gasping laugh, sweaty and tearful. It didn't stop when she asked the class to center ourselves with deep breathing. Just the suggestion struck me as hilarious. Then the ridiculous chortles that came from trying to stifle my laughter sent me over the edge.
That is when I noticed one or two of the participants possibly judging me. I imagined them after class whispering, "How about that conservative-looking 40-year-old? She was the looniest in the bin!" The thought brought me to near-passing-out laughter.
I drove home relaxed and exhausted. My abs were sore. I chuckled into the next day. My skeptical, introverted Marine reluctantly joined me for the next session. I watched and waited. He hit the tipping point at the 40 minute mark, startling me with the volume and intensity of his machine-gun laughter. Together, we lost it. Luckily it was laughing class; we were not asked to leave.
Laughter Yoga: Very good, very good YAY!
Contact the writer: firstname.lastname@example.com


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Create one-of-a-kind stretchy friendship bracelets
Kids will love creating stylish one-of-a-kind friendship bracelets with the FunLoom Kit. Bands are colorful and stretchy and can be used again and again! This craft kit may be used with other loom kits. For ages 5 years and up.
Fun Loom Kit Includes: 300 bands, hook, and loom
Basic Funloom Kit Dimensions: 13" x 5" x 3"
WARNING: CHOKING HAZARD-Small Parts. Keep away from children younger than 3 years.

subset: commoncrawl


text: It strains credulity to believe they created a mockup for their documentation to be used by end customers, when it would be simpler to take screen shots of the actual menus so that users would not be confused, should design challenges change the interface. As a former developer, I can tell you that it is far harder to document something that has yet to be written. While it is by no means impossible in this day of Photoshop and sophisticated editing software to create nearly anything imaginable, the question that would have to be asked is why a corporation would do this for one of their own products? At a public media event to demonstrate some new software or feature, sure, but to quietly slip it into their support pages without fanfare? Conspiracy theorists will never be satisfied, of course.


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Barbosa is a last-piece-of-the-puzzle type of player. We don't need to be wasting valuable cap space on an ageing 6th man, while the rest of the roster is still a complete unknown.
Yes, Calderon has arguably increased his trade value while logging heavy minutes this season, but unless a trade is actually consummated, increased trade value does nothing for the Raptors.
Im fine with Barbo getting signed long-term. 20mil for 4 years? too much. 2-3mil per year for 4-5yrs im ok with.
Oh, and as long as he doesnt play in the last 4 minutes of the 4th quarter. That offensive foul last night killed them.
And he's done that a couple of times in the past too.
I'm not a big fan of Barbosa. If he's hot he's a good scorer but he doesn't really bring anything else to the table and he's streaky. Hard to know what to think of an extension at this point. Depends on the money and length of the contract and it depends on what other options are out there. I'd rather see them move Barbosa for a younger player or pick(s) if they can.
and his demeanor is a very positive influence in the locker room, much like Calderon. As long as he doesnt go around sleeping with the other player's wives.
I would be upset if either Barbosa or Bayless were on the Raptors roster next season. They are both black-hole type players that don't understand the concept of team play. With a young core, those are not the values that veteran players should be instilling in the young players as examples of how to contribute to the team. Whatever scoring upside they might have is more than offset by their selfish, one-dimensional play. Trade them both at the deadline or let them walk in the offseason.
Although I do agree Bayless has problems finishing around the rim, I dont think that is a prerequisite for being a spark plug/6th man off the bench. He is very streaky just like Barbosa and I think he would actually be fine coming off the bench as a 6th man. Although all NBA players, especially younger players, strive to be a starter, Bayless seems to have accepted his role off the bench. He knows he cant complain about starting if he cant get onto the floor. For now he is still in a position where he needs to prove himself. If that means coming off the bench for a couple of years to prove yourself, most NBA players would take that and not complain.
For what it's worth I think within the next 1-2 years we will draft/sign/trade for Derozan's replacement and he will eventually be moved to the bench. Nothing against Derozan, he has shown some flashes over the last few games. On a contending team, which we all hope the Raps are in 2-3 years, I dont see him being more than a 6th or 7th guy, which is still a great role. For now i'm content with him occupying the 2 spot until we find a better alternative
i also agree with derozan not being good enough to start on playoff team too.. i hope im wrong on that though
besides BC has made it known that barbosa is one of his favorite players so if the money isnt too high i dont see BC letting him walk..
3 years and $9M would be good for me.
Include some incentives to get it to $12M over 3 years and I think that is very fair.
Personally I think he is just saying it so he won't get traded. Most players don't want to move somewhere where they have no choice. He was close to opting out of his contract last year to go back to Brazil to play. I think he's really not that interested in staying here. He rather just wait till the offseason.
Ryan Wolstat: Leandro Barbosa walked in to the ACC. I asked him if his Brazilian buddy Nene was signing with Toronto. "No man, it's too cold."
I wasn't a big fan of this comment, I think he should be recruiting guys a bit more.
I think you're being a little too sensitive. It could have been a joke. He could have been serious and that's what Nene told him. Either way I don't see the problem and I'm unsure how you can draw a line between that and him not trying to talk guys into playing in Toronto. It's not even his job to recruit. Management typically asks players to get involved like that and then it's usually the stars who are out there recruiting guys.
if they think its cold their a bunch of pussies thats all i can say.
I still think he will walk if we don't trade him. That's all I was saying is that he doesn't really care to be in Toronto but he doesn't want to be moved to location not of his choice for the next couple months. He is not a player that we are going to move forward with. I think there is places we can spend that money on. Unless we get him for a ridiculous price ( Matt's suggested 9 million over 3 years.).
Barbosa should be traded at the deadline for a young asset and/or a top 20 pick aka Kabongo
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Tim Burton is a genius! The darkness of all his movies brings a great feel to them. My favorite movie of his would either be Bettlejuice or The Nightmare Before Christmas.
Alice in Wonderland is definitely on my to see list when it comes out!
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MiharuSkii wrote:I can't wait for Alice in Wonderland! I just got done reading the book and I also just found out Alan Rickman is going to play the Caterpillar. +Johnny is going to be the Mad Hatter. It is so going to rock.
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Academically put, tropes are "common or overused theme[s] or device[s]," which makes them sound like cliches, which makes them seem like a bad thing.
But they're totally not! Romance novel fans all have their favorite—not to mention least favorite—tropes, from friends to lovers, chick in pants, secret baby, marriage of convenience, opposites attract, May-December, boss-assistant...the list goes on.
Each month, we'll be picking a romance novel trope and ask you to offer recommendations falling under the trope rubric (again with the academic talk!). This month, we want your recommendations for amnesia romances, or novels where either the hero or heroine has no clue who they are. As usual, we tried to remember (!) ourselves, but then went to our friends at Twitter.
Don't you forget about me...
- Sherry Thomas Tempting the Bride
- Lisa Kleypas Someone to Watch Over Me
- Miranda Neville Amorous Education of Celia Seaton
- Delilah Marvelle Forever and a Day
- Anne Stuart Winter's Edge
- Mary Balogh Slightly Sinful
- Anne Gracie Accidental Wedding
- Mary Jo Putney Uncommon Vows
- Beth Williamson The Redemption of Micah
- Author Vivian Arend shares that her forthcoming High Risk has a heroine who's missing some memories.
There are a lot of historicals on there—anyone got suggestions for the other genres? More historicals are welcome as well!


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Top prospects Taveras, Martinez back on track
By Jenifer Langosch / MLB.com
Two of the Cardinals' top three prospects are both moving in the right direction toward playing full-time in the Minors.
Oscar Taveras, the top prospect in the system and No. 3 on MLB.com's overall Top 100, is back with Triple-A Memphis. He made his 2013 debut on Monday as a pinch-hitter. He pinch-hit again the following day, leading some to wonder if there was a larger issue that was keeping him from being in the starting lineup after missing the start of the season to attend a funeral back home in the Dominican Republic and having travel issues getting back.
"There are no issues," Cardinals farm director John Vuch said about Taveras. "It's just that since he had been away for over a week without playing, arriving on Monday afternoon, we just wanted to give him a chance to get his legs back under him before throwing him out there for a full game. He's likely to be back in the lineup [Wednesday or Thursday]."
Right-hander Carlos Martinez, meanwhile, continues to get into game shape. The No. 3 prospect in the organization had visa trouble and couldn't get out of the Dominican Republic. He had his work visa approved on March 26 and made it to Florida two days later. It's now just a matter of time before he's deemed ready to head to Double-A Springfield.
"He'll be pitching at extended Spring Training for the next few weeks," Vuch said. "He'll likely be ready for activation by late April, early May."
-- Jonathan Mayo
Cardinals dedicate Friday's game to Musial
ST. LOUIS -- The Cardinals announced that they will dedicate Friday's game to honoring the life and legacy of Hall of Famer Stan Musial, who died in January at the age of 92.
Members of Musial's family will participate in a pregame ceremony, which will include a plaque dedication in Musial's honor. They will also throw out ceremonial first pitches. Rawlings has manufactured game balls stamped with a special Musial logo to be used during the Cardinals-Brewers contest.
Those attending Friday's game will receive a Stan Musial harmonica and commemorative box. Fans will then be asked to join in and play "Take Me Out to the Ballgame" -- a favorite of Musial's -- on the harmonica in the middle of the sixth inning, to match his uniform number. The organization will also distribute No. 6 window clings that resemble the Musial patch the Cardinals are wearing on their left sleeve throughout the season.
The Cardinals opened this homestand with a video tribute to Musial and the unveiling of a No. 6 decal on the wall in left-center field.
Carpenter, Descalso to split second base
ST. LOUIS -- With David Freese off the disabled list and presumably ready to resume playing almost daily, the Cardinals will be left to divvy up playing time at second base between Matt Carpenter and Daniel Descalso.
The pair entered Spring Training competing for the starting job at the position, though no clear winner emerged by the end of camp. Rather, manager Mike Matheny has said he intends to consider various factors on a daily basis to determine which of the two will start on any given day.
"There's going to be a juggling act a little bit," Matheny said.
The fact that Carpenter drew Wednesday's start behind sinkerball pitcher Jake Westbrook is further evidence of how far he has come defensively at the position. With the Cardinals believing Carpenter to present no defensive liability at second base, opposing pitchers are likely to play a large role in Matheny's daily decision.
Matheny also showed last year that he won't hesitate to stick for an extended period of time with whoever is rolling offensively. Carpenter entered Wednesday with multihit games in four of his seven starts. He ranked second in the league with four doubles and has scored in all five games in which he has gotten a hit.
Wednesday's start marked just his second of the season at second base. Descalso has started the other seven and is 7-for-28 at the plate.
"How well Matt has adapted to second base, I think, is very impressive," Matheny said. "It didn't surprise any of us. As soon as we started talking about it and put a work list together on the things he needed to work on in the winter, we knew it would probably be taken care of, and he did a great job."
Worth noting
• After playing every inning of the first eight games of the season, Allen Craig sat for Wednesday's series finale against the Reds. That gave manager Mike Matheny the opportunity to start first baseman Matt Adams, whose pinch-hit home run lifted the Cardinals to a win on Tuesday.
With Craig not starting, that leaves Yadier Molina, Jon Jay and Pete Kozma as the team's only position players to start all nine games this season.
• Despite appearing in each of the first two games of the series against the Reds, both Trevor Rosenthal and Mitchell Boggs were expected to be ready to pitch again on Wednesday, if needed. Their availability has a lot to do with both having low pitch counts on Tuesday. Rosenthal threw 11 pitches in the eighth inning; Boggs needed just 12 to cover the ninth.
• Right-hander Victor Marte has not had a smooth start to the season as Triple-A Memphis' closer. After blowing a save on Sunday, Marte again was unable to hold a ninth-inning lead on Tuesday. Memphis later lost the game, 5-4, in extra innings.
• In order to open a roster spot for Ryan Jackson in Triple-A, infielder Vance Albitz was sent down to Double-A Springfield.
Jenifer Langosch is a reporter for MLB.com. Read her blog, By Gosh, It's Langosch, and follow her on Twitter @LangoschMLB. This story was not subject to the approval of Major League Baseball or its clubs.


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|Número de publicación
|Tipo de publicación
|Número de solicitud
|Fecha de publicación
|29 Abr 1986
|Fecha de presentación
|18 Dic 1980
|Fecha de prioridad
|17 Oct 1978
|Número de publicación
|Lee R. Beck, Charles F. Flowers, Jr., Donald R. Cowsar, Albert C. Tanquary
|Cesionario original
|Stolle Research & Development Corporation
|Exportar cita
|BiBTeX, EndNote, RefMan
|Citas de patentes (12), Otras citas (26), Citada por (49), Clasificaciones (34), Eventos legales (7)
|Enlaces externos: USPTO, Cesión de USPTO, Espacenet
This is a continuation of application Ser. No. 952,109, filed Oct. 17, 1978, now abandoned.
1. Field of the Invention
The present invention relates to a method for introducing therapeutic or medicinal agents into the uterus and fallopian tubes. More particularly, the invention relates to a method of eliciting an active or passive immunization response in the internal female reproductive organs by introducing microparticles containing a specific antibody or antigen into the vagina and allowing the microparticles to be drawn through the cervix into the uterus.
2. Description of the Prior Art
In the past, the methods of generally treating the internal reproductive organs of the female have included principally the oral ingestion or the injection of drugs into the patient in order to treat diseases and to regulate the female reproductive cycle. No methods are known of treating the uterus and fallopian tubes by introducing a drug directly into the vagina where the natural transport mechanism of the internal reproductive organs conveys the drug across the cervix into the uterus. Yet, such a direct technique of locally introducing drugs into the vagina would be highly advantageous from the viewpoint of rapidly and effectively conveying drugs across the cervix into the uterus. This technique would be especially useful for the delivery of biologically active substances, such as antibodies and antigens, directly into the uterus to increase the level of immunization of the female reproductive organs. Because systemic antibodies are not secreted by the internal reproductive organs, the immunization levels of these organs cannot be increased by systemic administration of either antibodies or antigens, but rather must be increased by local administration of antibodies or antigens to the female reproductive organs.
In the past various drugs and cosmetic agents have been encapsulated in the form of microcapsules for the purpose of delivering these agents to the vagina by slow, sustained release of the agent from the microcapsules. However, these techniques have only been useful in the treatment of the vagina and not the other internal female reproductive organs. For example, Zaffaroni in U.S. Pat. No. 3,921,636 shows a drug delivery device in which microcapsules containing a medicinal agent are incorporated in a carrier device such as a tampon, sanitary napkin or intrauterine device. Thus, a tampon containing microencapsulated contraceptive hormone can be inserted into the vagina and the hormone will be gradually released by dissolution of the microcapsules. Since the hormone is released in the vagina, the vagina is the site in which the hormone is absorbed by the body. This technique does not provide a means of delivering drugs to the uterus by transport across the cervix.
U.S. Pat. No. 3,918,452 shows a technique in which a contraceptive agent is delivered to the vagina by inserting a tampon containing microcapsules composed of a contraceptive composition into the vagina. The contraceptive agent such as a spermicide is then released slowly with time into the vagina where the contraceptive agent has its effect. In this technique the effects of the contraceptive agent are limited only to the vagina and not to any other portions of the internal female reproductive organs.
Because it would be highly desirable to be able to introduce medicinal agents or therapeutic agents directly into the uterus and Fallopian tubes by transport of said agents across the cervix, various techniques have been attempt to achieve this end. One approach that has been suggested is to encapsulate a medicinal or therapeutic agent in the form of microcapsules and then deposit the microcapsules in the vagina whereupon the microcapsules are transported. As a result of some early investigations, it is known that carbon particles from a cap containing a suspension of carbon particles, when placed over the cervix, can be recovered from the uterus after coitus, as shown by Amersbach, "Sterilitat Und Frigiditat," Muchen. Med. Wchnschr. 77: 225, 1930. This shows that nonmotile particles migrate in the female reproductive tract. It was also demonstrated by J. Trapl, "Neuve Anschauunger uber den Ei-und Samentransport in den Geschlechtsteilen de Frau," Zentralbl. Gynak. 67: 547, 1943, that even without the use of a cervical cap, carmine particles migrate thus demonstrating that nonmotile particles other than carbon also migrate.
Still other investigators, R. Krehbiel and H. P. Carstens, "Roentgen Rabbit", Am. J. Physiol. 125: 571, 1959, have shown that the passage of a radio-opaque oil, when placed in the vagina of a rabbit was blocked until after the vulva was stimulated. Stimulation of the vulva caused contraction waves which transported the oil into the uterus and up into the uterolubal junction within several seconds. Other earlier investigations found that graphite and dyes in gelatin were not transported whether applied to the vagina before or after copulation, but carmine particles in cocoa butter were transported to the uterus and tubes. The implication of the data is that the nature of the particles affects the transport process and that transport is assisted by muscular contractions. Hartman, in "How Do Sperms Get Into the Uterus? Fertil. and Steril 8: 403, 1957, concluded that in the transport of sperm in the reproductive tract, transport occurs principally by cooperation of the particles with the musculature of the female reproductive tract. He also concluded that the function of the flagellum of the sperm is to aid in the penetration of the head of the sperm into the corona radiata, the zona pellucida and the vitelline membrane of the ovum. G. M. Duncan and D. R. Kalkwarf, "Sustained Release Systems for Fertility Control," in Human Reproduction: Conception and Contraception, edited by E. S. E. Hafez and T. N. Evans, Harper and Row, New York, 1973, have concluded from experiments that non-motile particles which are about the size of the head of the sperm migrate directionally through the cervix to the fallopian tubes. Thus, the reference indicates that non-motile particles of a size of 5 μm or less migrate throughout the internal reproductive organs when introduced into the vagina. However, when microcapsules of progesterone encapsulated within a suitable wall material such as cellulose acetate butyrate and of a size ranging from 5 to 1400 μm were introduced into the vagina, the microcapsules did not migrate across the cervix into the uterus, but rather were transported in the reverse direction. Therefore, the reference clearly suggests that microcapsules of a size greater than 5 μm will not migrate inward to the internal female reproductive organs.
A need therefore, continues to exist for a method by which various disorders and diseases of the internal female reproductive organs can be locally treated by applying microparticles of various medicinal and therapeutic agents to the vagina and allowing the natural transport mechanism of the organs to draw the microparticles across the cervix into the uterus where the medicinal or therapeutic agent is delivered to the uterus and other internal organs.
Accordingly, one object of the present invention is to provide a method by which medicinal and therapeutic agents can be locally administered to the vagina and transported through the cervix into the uterus to treat the internal female reproductive organs.
Another object of the present invention is to provide a method by which antigens or antibodies can be delivered to the uterus so that the internal organs can be treated directly to avoid systemic administration of the antigen or antibody.
Still another object of the present invention is to provide antibodies and antigens incorporated within microparticles which, when deposited in the vagina, can be transported across the the cervix into the uterus by the natural transport mechanism of the internal reproductive organs.
Briefly, these objects and other objects of the present invention as hereinafter will become more readily apparent can be attained by a method for achieving the passive immunization of the internal female reproductive organs by depositing antibody containing microparticles directly into the vagina and allowing the natural transport mechanism of the internal organs to convey the microparticles across the cervix into the uterus, whereby the antibody is continuously released from the microparticles.
The present invention can also be used to effect the active immunization of the internal organs by using microparticles which contain an antigen. The microparticles employed in the present process contain the antigen or antibody in a matrix which is biocompatible and biological degradable.
A more complete appreciation of the invention and many of the attendant advantages thereof will be readily obtained as the same becomes better understood by reference to the following detailed description when considered in connection with the accompanying drawings, wherein:
FIG. 1 shows microparticles of a monolithic structure containing a pharmaceutical agent;
FIG. 2 shows microparticles formed of a core of pharmaceutical agent in a matrix material surrounded by a shell of matrix material;
FIG. 3 shows microparticles formed of a core of pharmaceutical agent surrounded by a shell of matrix material;
FIG. 4 shows microparticles of an onion-skin structure of alternating layers of matrix material and pharmaceutical agent; and
FIG.5 shows microparticles formed of a core of one particular pharmaceutical agent surrounded by a shell of matrix material containing a second type of pharmaceutical agent.
The principal object of the present invention is to provide a method for delivering various antibodies or antigents directly to the internal reproductive organs to obviate systemic introduction of antigens or antibodies for the treatment of the reproductive organs. Systemic introduction, in fact, cannot be used as a means for administering many antigens and antibodies into the body for treatment of the reproductive organs. An antibody is defined as any body of globulins that combine specifically with antigens and neturalize toxins, agglutinate bacteria or cells, and precipitate soluble antigents. Antibodies are produced by the specialized cells of the endothelial reticular immune system in response to a challenge by antigens. The antibodies elicited by the challenge are highly specific to the antigen which evokes the response. This characteristic of antibodies has led to their exploitation in clinical medicine for both the diagnosis and treatment of disease. Moreover, both active and passive immunization have been used as a common therapeutic approach for the prevention and treatment of infectious diseases in man and animals.
In the past specific antibodies have been employed to retard the growth and proliferation of pathogenic microorganisms in both man and animals as well as to neutralize various bacterial toxins. Antibodies have also been used to control the growth and development of tumors and to modulate the immune system. Other medical uses of antibodies include the neutralization of the biological activity of drugs and hormones and localization of cells that convey unique antigens which may be associated with pathogenic conditions.
There are two basic ways in which the role of antibodies can be stimulated in the body to counteract the effects of antigens. One technique is active immunization while the other is passive immunization. In order to actively immunize a subject, the subject is administered an antigen to induce the formation of endogeneous antibodies. Normally, this technique requires up to two weeks before a sufficiently good level of antibody response is achieved. Because of the delay involved, the active immunization technique imposes limitations for the treatment of infectious diseases which have a short incubation time, for the treatment of a disease actively in progress and for reversing or modifying the effects of drugs, toxins, hormones, and enzymes. Furthermore, if active immunization is to be effective, the subject must have at least a functioning immune system which is capable of responding to the invading antigen. Thus, patients suffering from an immunodeficiency disease are precluded from active immunization. Yet a further restriction on the use of active immunization is that the antigens used to immunize a subject must be safe and non-toxic. The use, therefore, of toxic substances in the preparation of vaccines intended for human use is precluded.
The second basic immunization technique is passive immunization whereby antibodies are administered in order to achieve temporary immune protection. Passive immunization has the advantage that the biological effects are immediate and can be effectively used in patients suffering from immunodeficiency diseases. Moreover, active immunization is not limited to the use of non-toxic antigens because animal species can be used as the source for the protective antibodies.
The present invention provides a method of eliciting an active or passive immunization response in the internal reproductive organs of a female subject against an invading antigen as a result of the direct introduction of an antibody or antigen. The direct administration of these biologically active substances is achieved by incorporating specific antibody or antigen in microparticles and then introducing the microparticles of antibody or antigen into the vagina of a female subject and allowing the natural transport mechanisms of the internal organs involving in particular the cervix and uterus to convey the microparticles across the cervix into the uterus and eventually into the fallopian tubes. The microparticles release the antibody or antigen over a given period of time thus ensuring the presence of antibody or antigen in the reproduction organs which effectively provides for the active or passive immunization of the subject administered the microparticles.
In the technique of the present invention microparticles containing antigen or antibodies are deposited in the vagina. Transport of the microparticles is controlled by the cyclic changes in the endogenous ovarian steroid hormones, estradiol and progesterone. During the first 14 days of the menstrual cycle or the follicular or estrogenic phase of the cycle, the ovaries produce estradiol which has a stimulating effect on the cervical muscle contraction. The frequency and amplitude of the cervical contractions during the follicular phase steadily increase from day zero to day 14 at which time ovulation occurs and the cycle enters the lutial phase or progestational phase when the ovaries begin to secrete progesterone. Progesterone has an inhibitory effect on the contractile activity of the cervix or alternatively a muscle relaxing effect on the cervix. The ovarian hormones also exhibit an opposing effect in the cells of the cervix in that estradiol causes an accumulation of secretory products in the cells of the cervix while progesterone promotes the release of these products in the cervical lumen. The interactions described are the mechanism by which changes in the viscosity of the cervical fluid occur.
The fluid or mucous of the cervix is dynamic and is an aqueous type of hydrogel. The transport of microparticles as well as sperm is dependent upon the permeability of the cervical mucous to the microparticles as well as the propulsion provided by estrogen induced contractions of the cervix. The most appropriate time for transport of microparticles across the cervix occurs when the uterus exhibits maximum contractile activity and cervical mucous permeability. Accordingly, the greatest rate of transport of sperm or microparticles through the cervix should occur between day 12 and day 16 of the menstrual cycle, although the actual day of ovulation may vary from four to six days in different individuals with some transport occurring in some persons before day 12 of the cycle. Normally, the cervix is not very receptive to transport during the first twelve days of the menstrual cycle as well as between days 16 and 28. Thus, in order to deliver microparticles containing antigen or antibody into the cervix, the microparticles in an appropriate dosage need only to be deposited in the vagina prior to day 16 of the cycle, preferably before day 12 so that full advantage can be taken of the 4 day period at midcycle for transport of the microparticles across the cervix into the uterus.
In a major embodiment of the present invention advantage can be taken of the fact that estrogen and progestin hormones have a transport stimulating effect on the internal organs. Thus, microparticles containing either an estrogen or progestin or both can be introduced into the vagina to regulate the menstrual cycle of a woman who is not cycling regularly or to stimulate the cervix to transport microparticles across the cervix into the uterus and fallopian tubes. Once the microparticles containing cycle regulating hormone are deposited in the vagina, hormone is released by the microparticles and is absorbed by the vagina and/or in the cervix if indeed some microparticles have penetrated into the cervix. The locally absorbed hormone then induces the necessary secretory changes in the endometrium of the cervix and promotes the contractile activity of the cervix necessary for microparticles transport. Once transport activity has begun, the microparticles are conveyed across the cervix into the uterus. From the above discussion it is evident that microparticles containing cycle regulating hormone can be administered to regulate the menstrual cycle and to stimulate transport microparticles containing antigen or antibody across the cervix into the uterus.
The amount of estrogen or progestin administered via the microparticles should be an amount which, when delivered locally by absorption, is sufficient to regulate the menstrual cycle and to cause the change in the cervix which result in microparticle transport across the cervix. In other words, as the menstrual cycle regulating hormone is absorbed by the tissues of the vagina and indeed the tissues of the cervix if any microparticles have entered into the cervix, the desired regulatory effect is initiated immediately. The desired changes occur in the cervix which promote microparticles transport among which is a gradual increase in the contractile activity of the cervix. The administration of a large dose which results in achieving significant levels of the hormone on a systemic level, i.e. significant concentrations in the blood stream which result in delivery of the hormone to other organs such as the brain and liver in addition to the reproductive organs, should be avoided. Both estrogen and progestin in proper amounts will stimulate transport. However, a progestin at too high a level of concentration will have an adverse effect on microparticle transport because the progestins have a muscle relaxing effect on the tissues of the cervix. However, the adverse effect of the administered progestin can be reversed by the administration of a sufficient concentration of an estrogen via estrogen containing microparticles such as estradiol which as discussed supra induces contratile activity of the itssues of the cervix. Normally, when estradiol is delivered locally by way of the microparticles, the amount of microparticles administered should be sufficient to deliver from 0.1 to 1 mg per day over a 7-14 day period. On the other hand, when progesterone is incorporated in the microparticles, the amount of microparticles administered should be sufficient to deliver from 0.5 to 2 mg per day over a 7-14 day period.
As alluded to above the microparticles of the present invention can provide the feasibility of locally administering a contraceptive agent to the cervix while simultaneously exogenously activating a menstrual cycle in a non-cycling woman by the administration of appropriate ovarian hormone and stimulating the cervix for microparticle transport. Hence, estrogen containing microparticles which also contain the desired antibody or antigen agent can be administered such that estradiol or a synthetic estrogen is released at the cervix for a fourteen day period thus duplicating the first half of the menstrual cycle. When transport of the microparticles occurs across the cervix, antigen or antibody in the microparticles is delivered to the uterus. Fourteen days after administration of the estrogen containing microparticles, progesterone containing micrparticles optionally containing antibody or antigen are then administered. Thus, the complete natural menstrual cycle can be duplicated while providing antibody or antigen protection. It is also apparent that antigen or antibody and estrogen or progestin rather than being incorporated in the same microparticles can be incorporated in separate microparticles and delivered as a mixture so that each biologically active agent is present to deliver its intended function. Of course, it is also within the scope of this invention to deliver microparticles containing estrogen or progestin into the cervix to regulate the cycle and thereafter administer antigen or antibody containing microparticles at the period of the cycle when the cervix is receptive to transport. In the artificilly induced cycle maximum transport across the cervix is achieved between days 12 and 16. Since the cycle regulatory hormones are administered locally in the present technique, effective estradiol activity can be achieved at dosage rates between 0.01 and 0.07 mg per day, while effective progesterone activity can be attained at dosage rates of 0.04 to 0.14 mg per day. Estradiol and progesterone are the regulatory hormones of choice because they are naturally occurring endogenous hormones and therefore present no toxicity problems. However, it is evident that other well known synthetic estrogens and progestins can be employed as substitutes for estradiol and progesterone, respectively. Suitable estrogens include estrone, mestranol, ethinyl estradiol, 2-methoxyestrone, 2-hydroxyestrone and estriol. Suitable progestins include norethindrone, dimethisterone, ethynodiol diacetate, norethynodiol, norethindrone acetate and norgestrol. When the synthetic compounds are employed, the dose employed depends entirely upon the biological potency of the synthetic estrogen or progestin compound.
Microparticles containing antibody or antigen and/or menstrual cycle regulating hormone can be formed in a variety of configurations depending upon such factors as when during menstrual cycle the microparticles are delivered, whether sustained slow release or fast release of drug is desired, whether antigen or antibody is to be administered simultaneously with the menstrual cycle regulating hormone or after administration of the cycle regulating hormone, whether drug release is desired slowly and continually, intermittently or suddenly or the like. In perhaps the simplest situation as shown in FIG. 1 microparticles of a monolithic structure are prepared in which the desired antigen or antibody 2 is distributed throughout a matrix material 1 which is biodegradable and biocompatible. Once the microparticles are deposited in the vagina they begin to slowly deteriorate thereby continuously releasing the desired drug to achieve the desired daily dosage of drug over prolonged period of time from the time they are deposited in the vagina until well after the microparticles have been conveyed across the cervix and deposited in the uterus. In fact, the transport forces will result in some microparticles being deposited in the fallopian tubes. The term microparticles is used in a generic sense since the particles no matter in what particular configuration or regimen administered do not have to be spherically shaped in the form of microcapsules but can be of an irregular shape. Successful transport of the particles across the cervix is not dependent on the shape of the particles. When microparticles of a monolithic structure are administered, the drug diffuses out of the microparticles by gradual deterioration of the matrix material, by permeation of the drug out of the matrix, or by both mechanisms.
In another embodiment of the microparticle structure, as shown in FIG. 2, microparticles of a monolithic structure as shown in FIG. 1 are formed as described above. The monolithic microparticles are then further processed such that a wall or outer shell 3 of matrix material free of drug is formed on each microparticle. This type of microparticle configuration is desirable where release of the drug is to be delayed for some period of time after deposition of the microparticles in the vagina. The delayed release of drug obtained by using the above microparticles, for instance, would allow sufficient time for the microparticles to be deposited in the vagina, transported across the cervix and deposited in the uterus before the microparticles deteriorate to the point where the outer shell is essentially eliminated and drug release commences. While the thickness of the outer shell can be varied to any thickness desired, nevertheless, the overall size of the microparticles must be such that the microparticles possess sperm surrogate activity. If the microparticles are of such a relatively large size that they do not have sperm surrogate activity, then the microparticles will not be transported across the cervix and therefore cannot be used.
In still another embodiment of the microparticles as shown in FIG. 3, the microparticles can be designed for the sudden release of a large amount of antibody or antigen. To achieve this purpose the microparticles can be formed such that a core 5 of antigen or antibody is encapsulated in a shell matrix material 1. Microparticles containing a core of drug would be particularly well suited in situations where an endogenous factor for disrupting the outer shell of the microparticles is exploited. For example, the difference in pH of the mucosal fluids in the vagina on the one hand, and the cervix and uterus on the other hand, can be exploited such that deterioration of the outer shell occurs when the microparticles reach the area of the cervix or uterus. In this situation, the acidic pH of the vagina would have little or no effect on the shell of the microparticles. However, when the microparticles are conveyed into the cervix where they are exposed to the neutral pH therein, breakdown of the outer shell would commence eventually resulting in the sudden release of drug. This procedure would be particularly desirable where it is desired to administer a booster response after an individual has already received a primary immunization. Although it is more desirable to have a sustained release of antigen from microparticles of a monolithic structure, for instance, when an individual is to experience primary immunization, a shorter period of drug delivery is satisfactory for boosting the primary immune response.
With regard to passive immunization by the administration of antibody, it would be very desirable when a subject or patient has an acute infection or high concentration of toxin to be able to deliver a substantial amount of antibody quickly to the affected organ(s). By administering microparticles containing cores of antibody, once the microparticles are conveyed into and through the cervix, the antibody will suddenly be released in large quantities to counter the particular disorder. After the initial treatment microparticles could be administered to provide a sustained, lower level release of antibody to continue treatment.
In the treatment of patients for some disorders it is advantageous to be able to administer antigen or antibody in an intermittent fashion. This could be accomplished by the use of microparticles having the configuration shown in FIG. 4 where alternate layers of drug alone or dispersed in matrix material 7 and drug free matrix material 1 are formed in concentric layers. When such microparticles are deposited in the vagina, release of drug does not occur until the outer layer of the microparticles disintegrates. This could delay drug release until the microparticles are conveyed into the cervix and/or uterus. Once the underlying layer is exposed drug release starts and continues until the layer disintegrates or releases the drug. Drug release ceases as the next underlying drug free layer is reached. In this manner intermittent release of the drug is achieved. An example of the applicability of this technique can be found in active immunization where the outermost drug layer releases antigen for a sustained period which is followed by a period for instance of a week or two, in which no drug is released. After the non-drug containing layer disintegrates, a second period of antigen release starts. In this manner one could in a single administration of microparticles provide a primary immunization dose followed by a booster dose.
When it is desired to not only convey an antigen or antibody to the uterus through the cervix but also administer cycle regulatory hormone in order to activate or regulate the natural transport mechanism, it is possible to administer microparticles of a monolithic structure as shown in FIG. 1 in which both antigen or antibody and cycle regulating hormone are dispersed through a matrix material. In this manner, once the microparticles are deposited in the vagina, release of both hormone and antigen or antibody starts and eventually the microparticles are conveyed across the cervix into the uterus. A perhaps more selective regimen of administration could be provided by microparticles which have an outermost matrix layer containing cycle regulatory hormone and an inner core of matrix material containing antigen or antibody. When such microparticles are administered, the sustained release of cycle regulatory hormone occurs, and when the cervix is receptive to transport, the microparticles are transported across the cervix into the uterus. Release of the antigen or antibody will occur in the cervix or uterus as the underlying antigen or antibody core of the microparticles is exposed. FIG. 5 shows microparticles of the structure discussed above in which outer cycle regulatory hormone containing layer 9 encapsulates inner antigen or antibody containing core 11. The antigen or antibody alone can constitute the core of the microparticles or the antibody or antigen can be dispersed in matrix material to form core 11. However, outer layer or shell 9 is formulated by dispersing a menstrual cycle regulatory hormone in a matrix material.
From the above discussion it is evident that antibody or antigen alone or in combination with a cycle regulatory hormone can be incorporated in microparticles in a variety of configurations depending upon how the drug or drugs are to be released. Moreover, while multi-layered microparticles such as the types shown in FIGS. 2, 4 and 5 are normally formed of a single type of matrix material, it is possible, if not desirable under some circumstances, to formulate contiguous layers of the microparticles from different matrix materials. Still further it is possible that under some circumstances, it may be desirable to deliver more than one antibody or antigen to the internal reproductive organs to treat more than one condition. Thus, for instance, monolithic microparticles could be prepared and delivered containing two different antibodies to passively treat two different diseases. In fact, it may be desirable under some circumstances to actively immunize a patient against one disorder and simultaneously passively immunize the patient against a second disorder with antigen and antibody delivered in the same microparticles. Of course, when more than one antigen and/or antibody is combined in one microparticle where they may be in contact and not in different layers of a microparticle, they must not react with each other. A very similar situation exists where it may be desirable to include more than one type of estrogen or progestin in the same microparticle. The above embodiments only describe several of many possible microparticle configurations.
With regard to the physical size or shape of the microparticles, the microparticles can assume any possible shape ranging from ordered shapes such as of spherical or oval to irregular shapes. The shape of the microparticles is not a factor in microparticle transport.
The size of the microparticles is important insofar as the microparticles must possess sperm surrogate activity such that they can be conveyed by the natural transport mechanism of the reproductive organs upward from the cervix into the uterus and eventually by the fallopian tubes. If the microparticles are too large, they will cause contractions of the cervix which will expel the microparticles. Microparticles which are too small will not be conveyed upward into the internal reproductive organs. Usually, the microparticles range from 20 to 70μm, preferably 20-60μm. While the mechanism of microparticle transport is not known precisely, several factors are known which are important for transport. The cervical mucous is a type of aqueous hydrogel which is dynamic in that the viscosity and flow characteristics range during the monthly cycle of the reproductive organs. If transport of the microparticles is to be achieved, the matrix material of the microparticles must be compatible with the cervical mucous. The compatibility of the microparticles is a function of not only the chemical nature of the matrix material of the microparticles, but also the size of the microparticles. The microparticles must be miscible with the cervical mucous. Transport of the microparticles across the cervix, just as in the case of the sperm, is achieved only when the cervix is prepared to transport the microcapsules. Accordingly, the microparticles containing antigen or antibody must be deposited in the vagina when the cervix is ready to transport the microparticles. Once the cervix is ready for transport, syncronized muscle contractions of the cervis propel the microparticles into the uterus where eventually some of the microparticles are conveyed through the uterus into the fallopian tubes, although the important objective of the invention is that the microparticles should cross through the cervix into the uterus. The muscle contractions of the cervix are controlled by the natural release of hormones at the appropriate time of the monthly female cycle which as disucussed above is at midcycle. Alternatively, as discussed at length above, the transport mechanism can be stimulated by the administration of cycle regulatory hormone.
In the preparation of the antibody or antigen containing microparticles essentially any known antigen or antibody can be incorporated in the microparticles although those of particular use in the treatment of conditions and diseases of the internal reproductive organs are preferably used. Antibodies of the same type have the same biochemical structure regardless of what antigen they react with. Therefore, the same process can be used to incorporate any type of antibody in microparticles regardless of their specificity. Suitable types of antigens which can be incorporated in the present microparticles include bacterial and viral pathogens of man and animals, however, enzymes and other biological factors involved in the reproductive process can also be used.
Suitable examples of hormones acting as antigens include human chorionic gonadotrophin hormones, human placental lactogen, progesterone, estradiol and the like. Suitable antigens include those known as embryonic cellular antigens which occur on the cellular surface of the trophoblast and are unique to the trophoblast. In addition to the above mentioned pathogens, mixture of pathogens which can infect the female reproductive organs also can be incorporated in microparticles. Stated simply any pathogen can be used to produce a vaccine which could then be used to immunize the cervix, uterus and fallopian tubes by the present technique.
Examples of antibodies for passive immunization which can be incorporated in microparticles include those which correspond to all of the above described antigens which are effective for active immunization.
When the microparticles of the present invention are administered to a subject, they are administered in an amount such that the desired daily dosage level of antigen or antibody is delivered in an amount sufficient to elicit the desired response over the desired period of time. For treatment of the cervix, uterus and/or fallopian tubes a suitable dose range for an antigen for the role of primary active immunization would be 0.5 to 1 mg or antigen per day over a 7-14 day period. The dosage range required for a booster immunization would vary from 0.5 to 1 mg per day over a 24 hour time span. With regard to passive immunization via antibody administration, the weight of antibody administered does not necessarily directly relate to the therapeutic effect realized. The important factor in terms of dosage for passive immunization is the titer of the antibody or the biological potency. The titer of an antibody refers to the maximum dilution of the antibody which elicits an effect in a test situation. Two different preparations of antibody are not equally comparable on a weight basis because they have different biological potencies. An immunological titer of 1:500 is the minimum biological potency for any antibody to be administered by the process of the present invention. Moreover, the rate at which the immunoglobulin or antibody should be delivered to the cervix, uterus and fallopian tubes should not exceed 0.1 mg of antibody per day. Any dose rate less than this level which is effective in eliciting a therapeutic response is acceptable. Dosage rates greater than this level are unacceptable because the larger dose may cause a sensitization reaction against the antibody. That is, if the dosage rate is too great, the passively administered antibody might function as an antigen thereby stimulating the production of antibodies in the host that would react with the administered antibodies.
As described above, the matrix material in which the antigen or antibody is incorporated is an important consideration. Three factors predominate in the selection of a matrix material which are the biocompatibility of the matrix material with the mucosal fluids, the permeability of the matrix material and the ability of the matrix material to degrade biologically by a mechanism such as hydrolysis so that no matrix material residues remain after transport and deterioration of the microparticles in the uterus and fallopian tubes. The release of antigen or antibody from the microparticles may occur by diffusion of the agent through the entrapping matrix material or by erosion of the matrix material or by a combination of both factors. Suitable polymers as matrix materials include polyglycolic acid, polylactic acid, as well as copolymers of glycolic and lactic acid, and glycerol mono- and distearate. The preferred matrix materials, however, are polylactic acid and polyglycolic acid. The aliphatic polyesters described above degrade biologically by hydrolysis under physiological conditions and are converted to monomeric glycolic and lactic acids. The rate of degradation of the polymer in the body preferably occurs as soon as possible after the drug is released and is related to the rate of hydrolysis of the ester linkages which is, in turn, related to the surface area of the microcapsule or device, the crystallinity of the polymer, and the inherent hydraulic stability of the polyester.
The molecular weight of the particular polymer chosen is not a critical factor in the manufacture and use of the microcapsules. While an increase in the molecular weight of the polymer may gradually retard the rate of release of therapeutic agent(s) from the microcapsules and thus affect the dosage level administered, these factors may easily be compensated for by determining the rate of release of therapeutic agent(s) from the microparticles by in vivo or in vitro measurements and then adjusting the amount of microparticles administered in view of the results obtained from the release measurements.
The antigen or antibody containing microcapsules can be conveniently prepared by any well known procedure used in the past for the preparation of microparticles containing a pharmaceutical material. While the amount of antigen or antibody, and cycle regulatory hormone, if it is to be present, is not critical, normally, the microparticles contain from about 10 wt. % to 60 wt. %, preferably 10 wt. % to 50 wt. %, most preferably 10 wt. % to 25 wt. % of antibody or antigen. The methods selected for preparaing the microparticles are not critical although they will vary principally depending upon the type of microparticles to be prepared, i.e. whether the microparticles are to be monolithic, or manufactured such that a core of pharmaceutical material is surrounded by a wall of encapsulated matrix material or manufactured in a manner such that an onion-skin type of structure results with alternating layers of matrix material and pharmaceutical material alone or in a matrix material.
In the manufacture of the microparticles containing antigen or antibody and/or a menstrual cycle regulatory hormone, any conventional method of forming the microparticles can be used. The selection of a particular method chiefly depends upon the technical requirements of the matrix material and the particular manner in which the microparticles are intended to be used.
Generally, microencapsulation processes can be classified according to the three principal types of: (1) phase-separation methods including aqueous and organic phase separation processes, melt dispersion and spray drying; (2) interfacial reactions including interfacial polymerization, in situ polymerization and chemical vapor deposition; and (3) physical methods, including fluidized-bed spray coating, multi- and single-orifice centrifugal coating, electrostatic coating and physical vapor deposition.
Phase separation methods, as the term implies, rely on differential solubility characteristics that cause a wall- or shell-forming matrix material to separate from solution or suspension and deposit around particles or droplets of the therapeutic agent(s) to be encapsulated. The separation itself may be brought about physically, as by the addition of a non-solvent or by a change in temperature, or chemically, as by a change in pH.
An organic phase-separation process usually employs a dispersion or an emulsion of the therapeutic agent(s) in a solution or a high-molecular-weight polymer in an organic solvent. To this mixture is added a non-solvent or liquid polymer that causes the high-molecular-weight polymer to separate from solution and collect as a shell around the suspended therapeutic agent(s). The shell, still swollen with solvent, is then hardened by a further addition of non-solvent or by some other process that strengthens the shell and improves the barrier properties.
Typically, an aqueous solution or suspension of a lipophobic antigen or antibody and/or menstrual cycle regulatory hormone is added to a non-aqueous solution of a suitable matrix polymer, and the mixture is agitated to cause the formation of a water-in-oil emulsion. Depending upon its solubility in water, the agent may be present at a concentration of 5 to 50% in the aqueous phase, which may be 5 to 20% by weight of the total mixture. The external organic phase may contain 5 to 10% of the matrix polymer. Usually, however, the ratio of agent in the internal phase (aqueous solution or suspension) to polymer is 2:1 to 1:4. The polymer must be a good film-former; that is, it must possess adequate strength and toughness.
An aqueous phase separation process employs a dispersion or an emulsion of a water-insoluble therapeutic agent(s) in an aqueous solution of dispersion of a polymer. The polymer is caused to separate as gel particles; these collect around the therapeutic agent to form a shell; the shell is hardened; and the microparticles are isolated. In the coacervation process, which is the most common of the aqueous phase-separation processes, the water-insoluble therapeutic agent, whih may be in the form or particles or droplets, is usually dispersed in an aqueous sol of a hydrophilic colloid which becomes ionized in water; a second sol of opposite charge is added; and the mixture is caused to gel by a dilution with water, an addition of salt, an adjustment of pH, or a change in temperature, or by a combination of these. Appropriate conditions of coacervation are usually determined experimentally, because the various polymers, possible for use, differ significantly in physical and chemical properties according to source and method of isolation or preparation. A region of coacervation is determined by combining solutions or sols of two polymers at various concentrations, temperatures or levels of pH, and observing the conditions required for gelation. From these determinations can be drawn a ternary phase diagram, showing the area of compatibility and the region of coacervation, at a given temperature and pH. The changes in concentration, temperature or pH to effect gelation will then become apparent.
Each preparation of micriparticles requires carful control of conditions, and somehwhat different conditions are required for various therapeutic agents. The degree of agitation, for example, affects the size of emulsion droplets, and the surface properties of the droplets may require alterations in the procedures to insure deposition of matrix material about the droplets and to minimize formation of particles not participating in microencapsulation. The volume of water added in the dilution step is not critical, but generally larger volumes are required to maintain a stable emulsion when larger droplets are encapsulated.
The above phase separation can be adapted to an alternate technique in which the first step of forming a stable emulsion or suspension of the medicinal or therapeutic agent is accomplished by dispersing the agent in a solution of the matrix material. Thereafter, the emulsion is added dropwise to a non-solvent with stirring to precipitate the polymer coating material to form microparticles.
Another type of phase sparation technique is the melt-dispersion microencapsulation technique. This method can be used with a wide variety of medicinal or therapeutic agents. Usually a heat-liquefiable, waxy coating material, preferably of a low-melting wax such as glycerol distearate is suspended in an inert liquid such as a silicone oil or a fluorocarbon in which neither the wax nor the material to be encapsulated is appreciably soluble. The mixture is heated and stirred vigorously to melt and emulsify the wax. The therapeutic agent which has been powdered and screened to the desired size range, and the waxy coating material are dispersed with high shear agitation, and the liquefied wax coats the therapeutic agent to form the waxy liquid-coated microparticles. Thereafter, the formed microparticles are solidified by continued agitation which cools the particles. The microparticles are then isolated by filtration and dried as described earlier.
The second major method of forming the microparticles is by interfacial microencapsulation which involves bringing two reactants together at a reaction interface where polycondensation of the reactants, usually monomers, occurs to form a thin, insoluble polymeric film. One technique of establishing the interface for the encapsulation process is the dispersion or emulsification of the therapeutic agent with one of the reactants which form the condensation polymer in a continuous phase containing the second reactants.
The third major category of encapsulation techniques which is especially applicable to a variety of medicinal or therapeutic agents and coating materials is physical microencapsulation. The physical microencapsulation techniques are characterized by the continuous envelopment of particles or droplets of a medicinal or therapeutic agent in a fluid film, as a melt or solution of the coating material, in an apparatus containing coaxially or sequentially-spaced orifices. Thereafter, the fluid coating is hardened by a standard cooling technique or by solvent evaporation.
Among the physical methods for microencapsulation are those that involve the passage of liquid or solid core material through a liquid matrix material. The stream is disrupted by some means to cause the formation of liquid-coated droplets or particles, and the resulting particles are cooled or otherwise treated to solidify the shell material. For example, an aqueous solution of a therapeutic agent is aspirated into a rapidly flowing stream of molten glycerol distearate, and the mixture is ejected through a fine nozzle. On emergence from the nozzle, the liquid stream disintegrates into droplets, each consisting of an aqueous core surrounded by liquid wax. As these fall through air, the shells cool and solidify, and microparticles result. In another version of this process, the impelling force is supplied by a rotating member, which ejects the core material centrifugally through the shell-forming liquid.
The variations of these and other processes of microencapsulation are many. As is readily apparent to those skilled in the art, no one process nor any single set of conditions is applicable to all therapeutic agents, but instead a useful process is chosen and the conditions optimized to achieve the desired results with a specific agent.
Microcapsules containing medicinal or therapeutic agents can be delivered to the vagina by a variety of methods. The preferred method is to incorporate a fixed number of microcapsules into a container designed for easy hand insertion into the vagina. The insertion container should be made of a biodegradable material that dissolves within minutes after placement in the vagina, thus, releasing the microcapsules. Pharmaceutical type gelatin capsules can be conveniently used as a delivery system for the microcapsules. The dose level can be varied by increasing or decreasing the number of microcapsules in the delivery device. Of course, any number of other methods of variations, or this preferred method might be used. For example the microcapsules could be molded into a solid vaginal suppository by using an appropriate suspension medium such as gelatin. Creams, jellies, foams, or liquids might be used as a suspension medium for microcapsules. Preparations of this type could be placed in the vagina using a loadable syringe or some type of pressurized vaginal inserter. A variety of different types of applicators for administering pharmaceutical agents to the vagina and rectum are in common use. These consist of two parts; a nozzle design for easy insertion into the vagina, and a hand held implement used to project the preparation into the vagina. Syringes, squeeze bulbs, squeeze tubes and aerosol containers are examples of implements that can be used to generate the force necessary to propel the preparation into the vagina.
Vaginal suppositories offer the simplest, most direct method of application. The microparticles are inserted into the lower half of a preformed gelatin shell. The margin of the shell is then moistened with water and the upper half of the shell is joined to the lower half to complete formation of the suppository. Four, eight and twelve grain gelatin capsules can be used in this manner depending upon the dosage of microcapsules desired. Suppositories of other materials such a jellies, creams, foams or aerosols can also be used as the delivery system for microcapsules. The dose can be strictly regulated by including a fixed number of microcapsules in the suppository preparation, and the carrier device can be applied by hand. Another advantage is that by using a hollow container such as a gelatin capsule, special suspension media which might adversely affect the migration of the microcapsules are not needed.
The primary limitation for the generation of passive immunization in a subject by the administration of antibodies in clinical medicine is that antibodies produced in animals quite often cause serum sickness or anaphylaxis when injected into human recipients. However, the local delivery technique of the present invention in which microencapsulated antibodies are transported into the internal female reproductive organs circumvents this problems because not only are smaller dosages of antibodies required, but also systemic administration of antibodies is avoided. It is acknowledged that the use of humans as antibody donors can circumvent this problem. However, human donors cannot be used safely when immunization affects their own physiology or necessitate the use of antigens which are toxic. A distinguishing feature between active and passive immunization is that the natural elimination of passively administered antibodies from a subject renders this approach temporary and reversible, whereas active immunization of a subject is usually permanent and non-reversible.
Any type of reaction between administered antibody and antigen within the local environment of the internal reproductive organs to elicit a passive immunization response is within the scope of the present invention. For example, antibodies effective against any type of bacterial or viral pathogen can be used in the local treatment of infections in the vagina, cervix, uterus and fallopian tubes. Similarly, antibodies produced against sperm, egg, products of conception and biological factors in the reproductive fluids including hormones such as HCG and enzymes can be employed to prevent pregnancy.
Still another aspect of passive immunization within the scope of the present invention is the use of antibodies to reduce female fertility. The human preimplantation embryo produces and secretes a hormone called chorionic gonadotrophin hormone (HCG), which is necessary for implantation of the embryo into the uterus. It is known that antibodies against HCG neutralize the functions of this hormone and prevent implantation of the embryo from occuring. Accordingly, antibodies effective against HCG can be introduced into the internal female reproductive organs by the the technique of the present invention to prevent pregnancy. This technique can be extrapolated to the use of antibodies against sperm, egg, products of conception and a wide variety of enzymes and hormones which can be found in the fluids of the reproductive tract.
In some instances active immunization is more advantageous than passive immunization with an obvious example being active immunization for permanent protection against infectious diseases. Thus, when an antigen is incorporated within microparticles which can be delivered to the cervix, uterus and fallopian tubes, the delivered antigen induces the formation and secretion of specific antibodies by these organs. The secreted antibodies not only provide the desired immunological effect, but also are structurally and fundamentally unique from the type of antibody produced in response to systemic immunization. Systemic antibodies are not secreted by the reproductive organs, and it is for this reason that systemic immunization is not an effective way of generating antibodies in the fluids of the cervix, uterus and fallopian tubes. Thus, standard methods of immunization are not effective in the prevention of infections of the internal female organs or for controlling fertility. In the present invention, on the other hand, not only are antigens delivered directly to the internal female organs, but the mode of release of antigen from the microparticles varying from a sudden release to a sustained release ensures that the antigen can be administered as desired thus ensuring sensitization.
In a typical example of active immunization, antigens from the microorganism, Neisseria gonorrhea, for instance, are incorporated in a polymer such as polylactic acid. The microparticles are then administered into the vagina of a subject where they are then conveyed across the cervix into the uterus. The microparticles release antigen at the desired rate perhaps in the cervix as the microparticles are conveyed into the uterus. The antigen sensitizes the secretory tissues of the internal organs which respond by producing protective antibodies. The secreted antibodies form a protective fluid coating along the surfaces of the internal organs which protects the subject against an invasion and infection of N. gonorrhea microorganisms. The same technique can be employed to immunize a subject against other bacterial and viral infections of the internal female reproductive organs.
Another aspect of active immunization pertains to fertility. In this case, sperm antigens are delivered by transport of antigen containing microcapsules into the cervix, uterus and fallopian tubes. The antigen which is slowly released over a sustained period of time, stimulates the secretory tissues of the organs to secrete protective antibodies in the fluid layer which coats the internal organs which essentially are the cervix, uterus and fallopian tubes. After copulation and deposition of sperm in the vagina, antibodies in the cervical mucous cause agglutination of the sperm in the cervix and prevent further transport of the sperm into the uterus. Antibodies against sperm also inactivate sperm by techniques other than agglutination.
There are many recognized advantages to fertility control by immunization. The most obvious benefit is that immunization with an anti-fertility vaccine could provide sustained fertility control. An important advantage of the local administration technique of the present invention is that the vaccine could be self-administered at low cost. Recently, several antigens have been isolated and identified which are unique to the reproductive process which will induce an anti-fertility immune response. These antigens include those of the blastocyst, the ovum, the sperm, non-hormonal placental antigens and trophoblastic hormonal antigens.
The present concept of basing fertility control upon the local and direct administration of antigens to the internal female reproductive organs is founded on the assumption that a high concentration of antibodies within the reproductive tract may be more efficacious and safe for inhibition of sperm or blastocyst vitality than systemic immunization. It is believed that anti-sperm or anti-trophoblastic hormone immunity interferes with the reproductive process in the female genital tract before or during embryonic implantation. In fact, it is known that when HCG antigens are administered systemically, systemic antibodies may cross-react with other tissues of the body thereby giving rise to detrimental side-effects. The risk of the systemic side-effects precludes the use of this method for controlling fertility in humans. However, if indeed local immunity of the reproductive organs can be increased in the absence of a systemic immune response, then many of the complications associated with systemic immunization can be avoided. Furthermore, the present invention has the advantage that cyclic overdosing and underdosing which are inherent in conventional methods of administering drugs can be obviated by the sustained release of antigen or antibody from microparticles. Thus, the present technique affords a means for effecting a pharmacological response with a minimum dose of drugs.
Having now generally described the invention, a more complete understanding can be obtained by reference to certain specific examples which are included for purposes of illustration only and are not intended to be limiting unless otherwise specified.
A 2.5 g amount of progesterone and 10.0 g of d,l-polylactic acid were dissolved in 38 g of methylene chloride. The resulting viscous solution was poured into a 250 ml kettle containing 120 ml of a 5 wt. % aqueous polyvinylalcohol solution. The dispersion obtained was stirred at about 2000 rpm until a stable emulsion had formed with the droplets being in the range of 50 to 100 μm in diameter. A vacuum was applied to the emulsion until it began to foam and then the rate of stirring was reduced to 600 rpm. After two hours, most of the methylene chloride had evaporated. Moreover, continuous stirring was not required to prevent the embryonic microcapsules from agglomerating. Thereafter, the emulsion was centrifuged, the aqueous polyvinylalcohol solution was decanted and the microcapsules were resuspended in 150 ml of deionized water. For about 18 hours thereafter a vacuum was continually applied to the stirred aqueous suspension. Thereafter, the suspension was centrifuged and the microcapsules obtained were washed with water and then collected by vacuum filtration. The microcapsules were dried at room temperature under a hard vacuum overnight, and then they were sieved whereby a fraction ranging between 43 and 61 μm was obtained. By this procedure microcapsules containing 22±1.5 wt. % progesterone were obtained.
The procedure of Example 1 was followed to the extend that the ingredients were mixed and stirred in the aqueous polyvinylalcohol. A vacuum was applied to the stirred dispersion and after about 2 hours, when 90 wt. % of the solvent had been removed, the procedure was interrupted. The supension was centrifuged and the microcapsules were obtained after decantation. The microcapsules were resuspended in deionized water which did not contain a dispersing agent. A vacuum was reapplied to the suspended microcapsules and the procedure was continued to completion. By this technique the encapsulation efficiency was 100%.
A 1.0 g amount of progesterone was added to 4 g of molten glycerol monostearate and a portion of the molten mixture was poured into the reservoir of a melt sprayer and heated to 167° C. The flow of nitrogen into the device to effect cooling was 60 liters per minute, while the flow of nitrogen into the sprayer to aerosolize the molten mixture was adjusted to the maximum rate of 5.75 liters per minute. The aerosol was sprayed intermittently, and microcapsules were collected and sieved, whereby a size fraction ranging between 43 and 61 μm was collected. The microcapsules produced by this procedure were spherical and contained a 20 wt. % theoretical loading of progesterone.
Having now fully described this invention, it will be apparent to one of ordinary skill in the art that many changes and modifications can be made thereto without departing from the spirit or scope of the invention as set forth herein.
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|Clasificación de EE.UU.
|424/196.11, 514/899, 514/932, 514/931, 424/280.1, 424/193.1, 424/197.11, 514/963
|Clasificación internacional
|Clasificación cooperativa
|Clasificación europea
|27 Sep 1989
|Fee payment
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|1 Sep 1993
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|6 May 1996
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text:
What was decided under the North American Free Trade Agreement?
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In one sense, the answer to this question is very simple. In another sense, it is complicated and is far beyond the scope of an answer on this site.
Put simply, the North American Free Trade Agreement (NAFTA) created a free trade zone consisting of the three countries, the United States, Mexico, and Canada, that make up North America. A free trade zone is a zone in which there will be no barriers to trade between nations. Many nations set up trade barriers between themselves and other countries. For example, they might impose tariffs, which are taxes on imports that are coming into their country. As another example, they might impose quotas that limit the amount of some particular import that can enter their country. Economists argue that trade barriers are very harmful. They argue that countries will, on the whole, benefit when trade barriers are lowered. Because many government officials believe in this idea, NAFTA was approved by the governments of all three North American countries. In this view, then, NAFTA decided to tear down trade barriers between the US, Mexico, and Canada.
Looked at differently, the answer to this question would be huge. NAFTA is a tremendously detailed agreement that consists of more than 1,000 pages of terms. What this means is that the NAFTA negotiators had to decide on a myriad of issues. They had to decide on exactly how trade barriers would be taken down and by when this would have to happen. They had to decide if any exceptions would be made. They had to decide on how to enforce the treaty. In other words, whenever you have an agreement that runs over 1,000 pages, you have many things being decided. From this perspective, we can say that huge numbers of little details were decided by the NAFTA agreement.


text:
The Sox Move On Without Gentleman Jim and Jose
We knew "The Road Trip" was coming. We knew it would make or break the season. Still, it's hard to believe that our worst fears have come true and the Sox hopes for an October surprise are now awfully remote, if not next-to-impossible.
While it's not a complete surrender for 2009, the trades of Thome to the Dodgers and Contreras to the Rockies at the deadline are a signal that the Sox are ready to move on. It was unlikely Thome would be back next year anyway and the way Jose has performed recently his future wasn't on the South Side either. If you really think about it, if the rest of the club was playing well right now we could win the division without these two.
Thome is one of the great guys in the game and his numbers place him in a position to be among the greats in Cooperstown. That said, his almost four-year tenure with the Sox was a mixed bag. On one hand, we'll never forget his homer in the tiebreaker against the Twins last year to send us to the playoffs. And there were many other memorable blasts, including his 500th homer and subsequent ones that moved him up the HR ladder, but there were so many times he disappointed. As I've said in this space before, when a clutch double was needed he grounded out to short right field or took that all-too-familiar jaunt back to the dugout after a strikeout.
We'll remember Jose primarily for 2005: His 15-7 record with a fine 3.61 ERA in, his 17-game winning streak that spanned '05 and '06, his victories in the division series, LCS and World Series and his complete game in the LCS to send the Sox to the World Series–the fourth consecutive complete game by a Sox pitcher. But the sad fact is that it seemed Jose lost his confidence and simply became a detriment to the ball club. He only started against the Yankees last Saturday because there were no other viable options.
It's almost unfathomable to envision that the Sox, the way they are performing, will make a run the rest of the way. But I'll say it again: If the rest of the club starts playing up to expectations and goes on a roll there's no doubt in my mind that we can win without Thome and certainly without Contreras.