CH645545A5 - MICROPARTICLES FOR TREATING THE INTERNAL FEMALE GENESIS. - Google Patents

Description

The invention relates to microparticles for the treatment of the internal female genital organs.

The known methods for the general treatment of the internal female genital organs have mainly consisted of oral administration or an injection of medication into the patient for the treatment of diseases and regulation of the reproductive or menstrual cycle of the woman. Few methods are known for the treatment of the female genital organs by direct delivery of a therapeutic agent to the uterus. One method of delivering a contraceptive steroid hormone directly to the uterus is to use the progester device, a medicated intrauterine pessary. This device is known from US Pat. Nos. 3,699,951 and 3,777,015. This insertable device has the disadvantage that an experienced doctor is required for its introduction into the uterus. Accordingly, there is still a need for an improved method of delivering a contraceptive and other drugs such as antigens and antibodies to the internal female genital organs that allow self-introduction or self-administration of these agents.

Various methods are known for the introduction of contraceptives 35 in the form of microcapsules into the vagina, but in these methods the microcapsules are not transported into the uterus through the cervix. For example, U.S. Patents 3,918,452 and 3,921,636 disclose processes in which a pharmaceutical agent is released from microparticles contained in a tampon inserted into the vagina.

An intrauterine pessary is known from US Pat. Nos. 3,699,951 and 3,777,015 which is intended for the direct release 45 of progesterone into the uterus for contraception. However, this pessary is not biodegradable, and it must be brought into and removed from the uterus by an experienced doctor after use. The usability of this pessary is therefore limited by the fact that it cannot be introduced by the user.

Other publications that belong to the prior art are

U.S. Patents 3,773,919 ss 3,991,766

3976 071 3 691 271 3 887 699 3 490 454 and so 3 755 558,

as well as the publications Gardner, ACS Symposium, Series 33, 1976, pp. 171-181 and Yolles et al, ACS Symposium, Series 33, Chapters 5, 8 and 12, 1976.

Early studies were conducted to determine the area of the particulate matter for which it is possible to migrate through the cervix into the uterus after insertion into the vagina. It has been shown that carbon particles from one

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a suspension of carbon-containing cap-shaped pessary placed over the lower cervix after which coitus can be obtained from the uterus (Amersbach, “Sterility and Frigidity”, Münchn. Med. Wchnschr. 77: 225, 1930). J. Trapl has in "New Views on the egg and semen transport in the genitals of women", Zentralbl. Gynäk. 67: 547, 1943, showed that carmine particles migrate even without using a cap-shaped cervical pessary, thereby proving that other particles besides carbon, which are unable to move by themselves, migrate.

R. Krehbiel and H.P. Carstens in "Roentgen Rabbit", Am. J. Physiol. 125: 571, 1959 that the passage of an X-ray opaque oil into a rabbit's vagina was delayed until the external genitals were stimulated, while other researchers have shown that graphite and dyes in gelatin do not pass through the uterus are transported through. From this information it can be concluded that the transport process is influenced by the type of particles and that the transport is supported by muscle contractions. Hartman moved in "How Do Sperms Get Into the Uterus?" Fertile. and sterile. 8: 403, 1957 concluded that the sperm in the female genital organs are mainly promoted as a result of the interaction of the particles with the muscles of the genital organs. From experiments, G.M. Duncan and D.R. Kalkwarf ("Su-stained Release Systems for Fertility Control", in Human Reproduction: Conception and Contraception, edited by ESE Hafez and TN Evans, Harper and Row, New York, 1973) concluded that particles that are about the size of a sperm head and are not able to move on their own, migrate in a targeted manner through the cervix to the fallopian tubes. However, the article shows that progesterone-containing microcapsules made of cellulose acetate butyrate with a size between 5 and 1400 µm do not migrate through the cervix into the uterus, but are transported in the opposite direction. This reference therefore clearly suggests that microcapsules with a size of more than 5 μm do not migrate inwards to the internal female genital organs.

The object of the invention is accordingly to provide an agent in the form of microparticles with which active ingredients for treating the internal female genital organs can be administered locally into the vagina and transported through the cervix into the uterus through the natural transport mechanism of the inner genital organs.

This object is achieved by the microparticles defined in claim 1.

The supply of the active ingredients defined in claim 1 under a) and b) to the internal female genital organs is achieved by a use according to the invention in that microparticles containing the active ingredients are introduced directly into the vagina and in that the microparticles are passed through the natural one Transport mechanism of the internal genital organs through the cervix into the uterus.

The accompanying drawings are briefly explained below.

Fig. 1 shows monolithic microparticles containing the active ingredients.

2 shows microparticles which are formed from a core and a shell surrounding the core, the core consisting of the active ingredients in a matrix material and the shell consisting of matrix material.

3 shows microparticles which are formed from a core and a shell surrounding the core, the core consisting of the active substances and the shell consisting of matrix material.

FIG. 1 shows microparticles with an onion skin structure which have alternating layers of matrix material and of the active ingredients.

5 shows microparticles which are formed from a core and a shell surrounding the core, the core io consisting of a specific active substance and the shell consisting of a matrix material containing a second type of active substance.

FIG. 6 shows monolithic microparticles made from a contraceptive and a hormone used for regulating the cycle 15 in a matrix material.

7 shows microparticles consisting of a core and a shell surrounding the core, the core being composed of a contraceptive and a hormone which serves to regulate the cycle, and the shell is composed of matrix material.

8 shows microparticles consisting of a core and a shell surrounding the core, the core consisting of a contraceptive and a hormone used to regulate the cycle in a matrix material and the shell 25 consists of matrix material.

9 shows microparticles consisting of a core and a shell surrounding the core, the core consisting of a contraceptive agent in a matrix material and the shell consisting of a hormone 30 used for regulating the cycle in matrix material.

Fig. 10 shows multiple layers of microparticles in which a contraceptive and a cycle regulating hormone are completely dispersed in different layers.

35 Figures 11A and IIB show the blood levels of progesterone and estradiol in baboon females to whom 7.87 mg of progesterone were administered intramuscularly and intravaginally, respectively.

Figure 12 is a series of photomicrographs showing the histological appearance of the baboon endometrium.

Fig. 13 is a series of photomicrographs showing the morphology of the uterine epithelial surface of baboons.

45 Figures 14A and 14B show the blood levels of progesterone and estrogen in baboon females to whom 1.57 mg of progesterone were administered intramuscularly and intravaginally, respectively.

Figure 15 is a series of microphotographs showing the morphology of the baboon endometrial epithelial surface.

Figures 16A and 16B are records of the contractile activity of the cervix of two baboon females, one of which had been treated with 55 microparticles containing estradiol while the other was not treated with hormone-containing microparticles.

By introducing the microparticles according to the invention into the vagina, the active substances are conveyed through the natural transport mechanism through the cervix into the uterus and into the fallopian tube, where the active substances are released. In this way, various antibodies or antigens can be transported directly to the internal genital organs, making systemic treatment of the genital organs with antigens or antibodies unnecessary. In fact, for many antigens and antibodies, the introduction into the organism or systemic introduction is not a means

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their administration into the body for treatment of the genital organs in question.

By introducing microparticles that contain a contraceptive, the microparticles are transported through the cervix into the uterus. The direct and local introduction of the contraceptive has the advantage that much smaller doses of many types of contraceptive can be administered by yourself and that practically the same contraceptive effect is achieved as when introducing large amounts of a contraceptive into the organism. This method is used to deliver microparticles containing a contraceptive, normally, i.e. in a woman who has a regular menstrual cycle. The microparticles according to the invention can also be used to regulate the menstrual cycle of a woman who does not have a regular cycle.

At the same time, the microparticles have the effect that the internal genital organs are increasingly enabled to carry the microparticles containing a contraceptive agent through the cervix into the uterus and fallopian tubes. The desired effect is achieved by adding a hormone to the microparticles in addition to the contraceptive, which regulates the menstrual cycle and facilitates or promotes transportation through the uterus, this hormone generally being in estrogen or progestogen.

There are two basic methods by which the role of antibodies in the body can be stimulated to counteract the effects of antigens. One is active immunization and the other is passive immunization. To actively immunize a patient, an antigen is administered to the patient to induce the formation of endogenous antibodies. This procedure generally takes up to two weeks to achieve a satisfactory immune response from antibody formation. Because of the delay associated with this, the treatment of infectious diseases with a short incubation period, the treatment of diseases with an active course and the reversal or modification of the effects of drugs, drugs, narcotics, toxins, hormones and enzymes by means of active immunization are imposed. The second basic immunization procedure is passive immunization, in which antibodies are administered to achieve temporary immunity.

Passive immunization has the advantage that the biological effects are immediate and can be used effectively in patients suffering from immunodeficiency disorders. Furthermore, passive 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 transport of the microparticles is regulated by the cyclical changes in the endogenous ovarian steroid hormones, estradiol and progesterone. During the first 14 days of the menstrual cycle, the follicular maturation phase or the estrogen phase of the cycle, the ovaries produce estradiol, which has a stimulating effect on the contraction of the cervical muscle. The frequency and amplitude of the cervical contractions increase continuously during the follicular cleansing phase from the 0th to the 14th day. Ovulation occurs on the 14th day and the cycle enters the luteal or progesterone phase when the ovaries begin to excrete progesterone. Progesterone has an inhibitory effect on the contractile activity of the cervix or alternatively a relaxing or slackening effect on the muscle of the cervix. The ovarian hormones also show an opposite effect in the cervical cells, which is that estradiol causes secretory products to accumulate in the cervical cells, while progesterone promotes the release of these products into the cervical lumen. The described interactions io are the mechanism by which changes in the viscosity of the cervical mucus occur.

The fluid or mucus of the cervix (the cervical mucus) is dynamic and is an aqueous type of hydrogel. The transport of microparticles 15 and sperm depends on the permeability of the cervical mucus to the microparticles and on the driving force in the form of contractions of the cervix that are caused by estrogen. The most suitable time for the subsequent transportation of microparticles through the cervix occurs when the uterus has the maximum contractile activity and the maximum permeability of the cervical mucus to the microparticles. Thus, between the 12th and 16th days of the menstrual cycle, the rate of delivery of sperm or microparticles through the cervix should be greatest, although the actual day of ovulation may vary by 4 to 6 days in different individuals, so it may vary certain people before the 12th day and between the 16th and 28th 30 days of the menstrual cycle are not very susceptible to transportation. Therefore, in order to deliver microparticles containing antigens, antibodies or contraceptives into the cervix, the microparticles need only be inserted into the vagina in a suitable dose amount before the 16th, preferably before the 12th, day of the 35 cycle.

In a preferred embodiment of the invention, the fact can be exploited that the estrogen and progestogen hormones have a stimulating effect on the internal genital organs. For example, microparticles that contain either an estrogen or a progestin and a drug, or microparticles that contain an estrogen or progestogen, along with microparticles that contain a drug, can be inserted into the vagina to help regulate the menstrual cycle of a woman who is not regular Cycle, to regulate or to stimulate the cervix so that microparticles are transported through the cervix into the uterus and fallopian tubes. The locally absorbed gestagen or estrogen in such a biologically effective amount induces the necessary secretory changes in the endometrium of the cervix and supports or stimulates the contractile activity of the cervix, which is necessary for the transport of microparticles. Once the transport activity has started, the microparticles are transported through the cervix into the uterus. Transport is stimulated by both estrogen and progestogen in suitable amounts. Gestagen, which is used in a concentration too high, has an adverse effect on the transport of microparticles because the gestagens exert a muscle-relaxing or muscle relaxation effect on the tissues of the cervix. However, the adverse effects of the progestogen administered can be counteracted by administering a sufficient concentration of an estrogen, whereby the estrogen, e.g. Oestradiol, which, as explained above, is the contractile activity of the tissues

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of the cervix, is administered in the form of estrogen-containing microparticles. When estradiol is delivered locally using the microparticles, the amount of microparticles administered should generally be sufficient to deliver 0.1 to 1 mg per day over a period of 7 to 14 days. On the other hand, if progesterone is incorporated into the microparticles, the amount of microparticles administered should be sufficient to supply 0.5 to 2 mg per day over a period of 7 to 14 days.

As indicated above, the microparticles according to the invention offer the possibility of locally administering a contraceptive, an antigen or an antibody to the cervix, while at the same time activating the menstrual cycle of a woman, whose monthly menstrual period is absent, by administering a suitable ovarian hormone and activating the cervix is stimulated exogenously for the transport of microparticles. Thus, estrogen-containing microparticles can be administered by releasing estradiol or a synthetic estrogen on the cervix over a period of 14 days, mimicking the first half of the menstrual cycle. When the microparticles are transported through the cervix, a medicament present in the microparticles or in separate microparticles is supplied to the uterus. 14 days after the administration of the estrogen-containing microparticles, progestone-containing microparticles, which may contain an antibody or an antigen, are then administered. In this way, the entire natural menstrual cycle can be mimicked while offering protection by antibodies or antigen. It is of course also within the scope of the invention to deliver microparticles containing estrogen or progestin to the cervix to regulate the cycle and then to administer antigen or antibody containing microparticles in the cycle period in which the cervix is susceptible to delivery. In the artificially induced cycle, the maximum transport through the cervix is achieved between the 12th and 16th day. Since the cycle-regulating hormones are administered locally in the method according to the invention, an effective estradiol activity can be achieved with dose amounts between 0.01 and 0.07 mg per day, while an effective progesterone activity with dose amounts of 0.04 to 0. 14 mg a day can be achieved. Oestradiol and progesterone are the cycle regulating hormones of choice. However, other known synthetic estrogens and progestogens can of course also be used as replacements for estradiol or progesterone. Examples of suitable estrogens are estrone, mestra-nol, ethynyl estradiol, 2-methoxy estrone, 2-hydroxy estrone and estriol. Examples of suitable gestagens are noret-hindron, dimethisterone, ethynodiol diacetate, norethynodiol, norethindrone acetate and norgestrol. When the synthetic compounds are used, the dose used depends entirely on the biological effectiveness of the synthetic estrogen or progestogen compound.

The microparticles according to the invention can be produced in a large number of embodiments. Probably the simplest case, which is shown in FIGS. 1 and 6, monolithic microparticles are produced in which the desired antigen or the desired antibody 2 on the one hand or the contraceptive agent 21 and the cycle regulating hormone 22 on the other hand completely in one Matrix material 1 (or 23) that is biodegradable and biocompatible can be distributed. Once the microparticles have been inserted into the vagina, they begin to slowly decompose, causing the desired drug to extend over an extended period of time, from the time the microparticles are inserted into the vagina long after the microparticles are transported through the cervix and stored in the uterus, the desired daily dose is continuously released. When the microparticles contain a cycle-regulating hormone, the properties of the cervical mucus and the contractile activity of the cervix are regulated by the continuous release of the hormone. In this particular embodiment, since the microparticles continuously release their active ingredients from the time they are inserted into the vagina, it is obvious

that very favorable results are obtained if the microparticles are formed so that their effective lifespan is close to the intermenstrual area so that the microparticles need only be administered once during the cycle. However, this is only a preferred embodiment because the microparticles can be administered as many times as desired to achieve the contraceptive and cycle regulating effects.

FIG. 2 shows another embodiment of the microparticles with a monolithic structure in which an antigen or antibody 2 is incorporated into a matrix material 1, which in turn is surrounded by an envelope 3 made of a drug-free matrix material. This type of microparticle shape, in which the particles are sized to act as a sperm proxy, is desirable if the release of the drug is to be delayed for a period of time after the microparticles have been inserted into the vagina. For example, the delay in drug release achieved by using the microparticles described above would allow enough time to allow the microparticles to be stored in the vagina, passed through the cervix, and stored in the uterus before the microparticles are deposited decomposed to the point where the outer wall has essentially been removed and the drug begins to be released.

7 shows microparticles in which a core 25 made of a contraceptive agent 21 and a hormone 22 regulating the cycle is encapsulated in a shell 27 made of a matrix material 24. This particular shape of the microparticles would be desirable when it is necessary to delay the release of the active components of the supplied microparticles, as described above for the microparticles of FIG. 2.

In another shape of the microparticles shown in Fig. 3, the microparticles can be determined to suddenly release a large amount of the antibody or antigen. For this purpose, the microparticles can be formed in such a way that a core 5 made of antigen or antibody is encapsulated in a shell 1 made of a matrix material. In cases in which an endogenous factor is used to split or break the outer shell of the microparticles, microparticles which contain a core from a medicament would be particularly suitable. For example, the pH difference between the slimy fluid in the vagina on the one hand and in the cervix and the uterus on the other hand can be exploited in such a way that the outer shell is decomposed when the microparticles reach the area of the cervix or uterus. In this case, the acidic pH of the vagina would have little or no effect

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on the shell of the microparticles. However, the breakdown or cleavage of the outer shell would begin when the microparticles are transported to the cervix, where they are exposed to the neutral pH therein, and this would ultimately lead to the sudden release of the drug, which is advantageous if to deliver a significant amount of an antibody to a patient who has an acute infection or has a high concentration of a toxin. This method would be particularly desirable if one intends to cause a boosting refreshment response in an individual who has already received primary immunization.

When treating patients with certain health disorders, it is advantageous to be able to intermittently administer an antigen or an antibody. This could be achieved by using microparticles with the shape shown in FIG. 4, in which concentric layers are formed which alternately consist only of medicament or of medicament dispersed in a matrix material (layers 7) and of medicament-free matrix material (layers 1). consist. When such microparticles are inserted into the vagina, the drug is not released before the outer layer of the microparticles decompose. As soon as the layer beneath the outer layer is exposed or exposed, the release of the drug begins, which continues until the layer has decomposed or the drug has been released. The release of the drug stops when the next underlying drug-free layer is reached. In this way an intermittent release of the drug is achieved. An example of the applicability of this method can be found in active immunization, where the outermost drug layer releases antigen over a continuous period, to which a period of e.g. a week or two weeks later in which no drug is released. After the layer containing no drug has decomposed, a second period of antigen release begins. In this way, a single dose of microparticles could be used to provide a primary dose of immunization and a subsequent boost or boost dose.

If it is desired not only to deliver an antigen or an antibody to the uterus through the cervix, but also to administer a cycle regulating hormone to activate or regulate the natural transport mechanism, the administration of monolithic microparticles is possible, as shown in Fig. 1 are shown in which both antigen or antibody and a cycle regulating hormone are completely dispersed in a matrix material. In this way, the release of both hormone and antigen or antibody begins as soon as the microparticles are inserted into the vagina, and the microparticles are ultimately transported through the cervix into the uterus. A mode of administration with possibly even higher selectivity could be achieved by microparticles which have an outer matrix layer in which a cycle regulating hormone is contained and an inner core made of a matrix material containing an antigen or an antibody. When such microparticles are administered, the cycle regulating hormone is permanently released, and when the cervix is receptive to delivery, the microparticles are delivered through the cervix into the uterus. The antigen or antibody is released in the cervix or uterus when the core of the microparticles, which contains antigen or antibodies and is located under the outer matrix layer, is exposed or exposed. 5 shows microparticles with the structure explained above, in which core 11, which contains antigen or antibodies, is encapsulated by an outer layer 9, in which a hormone regulating the cycle is contained. The core of the microparticles can consist of the antigen or the antibody alone, or the antibody or the antigen can be dispersed in the matrix material to form the core 11. However, the outer layer or sheath 9 is formulated by dispersing a menstrual hormone regulating in a matrix material.

The foregoing clarifies that an antibody or antigen, alone or in conjunction with a cycle regulating hormone, can be incorporated into microparticles in a variety of configurations or structures depending on how the drug or drugs are to be released . Although the multi-layer microparticles such as the types shown in FIGS. 2, 4 and 5 are generally formed from a single type of matrix material, it is also possible, if desired under certain conditions, for mutually adjacent layers of the microparticles 25 formulated from different matrix materials. In addition, under certain circumstances it may be desirable to deliver more than one antibody or more than one antigen to the internal genital organs to treat more than one condition. For example, monolithic 30 microparticles are produced and supplied, which contain two different antibodies for the passive treatment of two different diseases. In fact, under certain circumstances, it may be desirable to use an antigen or antibody that is supplied in the same microparticles to immunize a patient simultaneously against a health disorder and passively against a second health disorder. Of course, if men as an antigen and / or antibody are combined in a microparticle where they can be in contact and if not in different layers of the microparticle, they must not react with each other.

Another microparticle structure is shown in Fig. 8, where a contraceptive 21 and a cycle regulating hormone 22 are compatible, i.e. 45 are not reactive with one another, are dispersed in a matrix material 23 to form a core 25. The microparticles are completed by encapsulating the particles of the drug-containing matrix material in a shell 27 made of a matrix material 24. The matrix materials 23 and so 24 of the inner core 25 and the sheath 27 can consist of the same material or of different materials. The use of different matrix materials is particularly useful when it is desired to take advantage of the different rates of decomposition of the matrix materials ss or the different rates of diffusion of the drug through the matrix materials. The contraceptive and the cycle regulating hormone are not released before the shell of the matrix material 24 is decomposed.

6o FIG. 9 shows a microparticle structure in which a core 25 is formed from a contraceptive 21 in a matrix material 23, which core is then surrounded by a shell 27 made of a matrix material 24, in which a hormone 22 regulating the cycle is contained. The microparticles with this particular structure are particularly suitable when it is desired to insert microparticles into the vagina in order to initially only gradually release the cycle regulating hormone 22, through which the

Regulates the monthly cycle and stimulates the transport of microparticles through the cervix. In this way, the regulation of the monthly cycle and an at least initial transport of the microparticles can be initiated at the point in time at which the inner core 25 is exposed enough to allow the encapsulated contraceptive 21 to be released. Of course, the contraceptive may be encapsulated alone or fully dispersed in a core matrix material that is the same as or different from the shell 24 matrix material.

10 shows a multi-layer microparticle structure. These microparticles have an inner core 25 made of a contraceptive agent 21, which is present alone or is dispersed in a matrix material 23. The core 25 is encapsulated in a shell 27 made of a matrix material 24, which in turn is encapsulated in an outer shell 31 made of a matrix material 29 which contains a hormone 28 which regulates the cycle. Microparticles with this particular structure would be desirable in cases where, after the microparticles have been inserted into the vagina, it is desired to deliver the cycle regulating hormone. After all of the cycle regulating hormone has been released, the release of the contraceptive would be delayed until the inner shell 27 has decomposed to a sufficient extent to allow the contraceptive to be released from the core 25. In the meantime, the microparticles are delivered so that a delayed release of the contraceptive is achieved until most of the microparticles have been delivered to the uterus.

As for the physical shape of the microparticles, they can take any shape, from regular to spherical or oval to irregular. The shape of the microparticles plays no role in their transportation. In general, the drug diffuses from the microparticles because of the gradual decomposition of the matrix material and / or because the drug penetrates through the matrix material.

The size of the microparticles is important insofar as the microparticles have to act as a proxy for the sperm, so that they can be transported upwards from the cervix into the uterus and finally or, if necessary, into the fallopian tubes through the natural transport mechanism of the genital organs. If the microparticles are too large, they cause contractions of the cervix, through which the microparticles are expelled or expelled. Microparticles that are too small are not transported upwards into the internal genital organs. The microparticles generally have a size between 20 and 70 μm, preferably between 20 and 60 (in.

The matrix material from which the microparticles are formed and in which the contraceptive and the cycle regulating hormone are dispersed is important not only from the point of view of acting as a sperm, but also in terms of biological compatibility. The material should not have an adverse effect on the internal female genital organs so that it is suitable as a matrix material. The matrix material should also be biocompatible in that it does not irritate the tissues of the cervix or uterus; it must not be carcinogenic and should not cause inflammation in the tissues of the body. The matrix material should be compatible with the body tissues and must be mixed with the cervical mucus645545

be cash. Another important factor is that the matrix material must be biodegradable in the sense that the body's chemical processes are able to ultimately degrade the polymer so that it does not accumulate in the body. The matrix material should also have the ability to slowly decompose in the form of the microparticle over a period of time at least equal to the intermenstrual vacuum. Examples of suitable polymeric materials are polymers or polycondensates of glycolic acid, polymers or polycondensates of d, l-lactic acid, their copolymers, etc. Examples of other suitable matrix materials are materials such as glycerol mono- and distearate. Examples of other matrix materials are materials that decompose in the neutral environment of the cervix.

In the production of the microparticles containing antibodies or antigens, essentially all known antigens or antibodies can be incorporated into the microparticles, but the antibodies or antigens which are used to a particular extent for the treatment of conditions and diseases of the internal genital organs are preferably used. Examples of suitable types of antigens which can be incorporated into the microparticles according to the invention are bacterial and viral pathogens of humans and animals, but enzymes and other biological factors which are connected to the reproductive process can also be used. Examples of suitable pathogenic antigens are Neisseria gonorrhea, Mycobacterium tuberculosis, herpes virus (hominis, types 1 and 2), Candida albicans, Candida tropicalis, Trichomonas vaginalis, Haemophilus vaginalis, Streptococcus coli group 'B, Microplasma hominis, Hemophiluloma ducreyi , Lymphopathia venereum, Treponema pallidum, Brucella abortus, Brucella melitensis, Brucella suis, Brucella canis, Campylobacter fetus, Campylobacter fetus intestinalis, Leptospira pomona, Listeria monocytogenes, Brucella ovis, Perde-Herpes-Virus 1, equine-virus-virus 1, equine-virus-virus 1 IBP virus, BVD-MB virus, Chlamydia psittaci, Trichomonas foetus, Toxoplasma gondii, Escherichia coli, Actinoba-cillus equuli, Salmonella abortus ovis, Salmonella abortus equi, Pseudomonas aeruginosa, Corynebacterium pyogenes, Actinobacillium semin, Actinobacillium semis Absidia ramosa, Trypanosoma equiper-dum, Babesia caballi, Clostridium tetani and Corynebacterium equi.

Suitable examples of enzymes that may be associated with the reproductive process are ribonuclease, neuramidinase, trypsin, glycogen phosphorylase, sperm lactate dehydrogenase, sperm hyaluronidase, ade nosin triphosphatase, alkaline phosphatase, alkaline phosphatophenase trypsinase, Amylase, muramidase, acrosome proteinase, diesterase, glutamic acid dehydrogenase, succinic acid dehydrogenase, ß-glycolophosphatase, lipase, ATP-ase-a-peptate-y-glutamylotranspeptidase, sterol-3-ß-ol-dehydrogenase and DPN-dihydrogenase aprorase.

Suitable examples of hormones which act as antigens are the gonadotropic chorionic hormones in humans, the placental tactogen in humans, progesterone, estradiol etc. Examples of other antigens are the antigens known under the name “embryonic cellular antigens” which are located on the cellular surface of the trophoblast and only with this occurrence. In addition to the pathogens mentioned above, mixtures of pathogens that can infect the female genital organs can also be incorporated into microparticles.

Examples of passive immunization antibodies that can be incorporated into microparticles are those

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which correspond to the antigens described above, which are not excreted by the genital organs, and which are effective for active immunization. For this reason too, systemic immunization is not an effective

Bodies that are effective against sperm, eggs, products of conception, fective method for the production of antibodies in etc. can be used. the fluids of the cervix, uterus and

If the antigen or antibody-containing micro- the fallopian tubes.

Another aspect of active immunization is administered in such an amount that the desired daily refers to fertility. In this case, the dosage level of the antigen or antibody is supplied in an amount by the transport of antigen-containing microcapsules which is sufficient to achieve the desired response or immune response over the desired period of the cervix, uterus and fallopian tubes. Antigens supplied. The antigen that evokes about you. This would be released slowly with an antigen of a long period of time, stimulating the secretion of about 0.5 to 1 mg of the antigen per day over a temporal tissue of the organs, so that in the fluid layer, space corresponds to 7 to 14 days . The dough required for a reinforcement to cover the internal organs, in which essential or repetitive immunization was required around the cervix, uterus and the area of application would be from 0.5 to 1 mg per day via a 15 fallopian tube protective antibodies excreted who vary the period of 24 h. With regard to passive imens. After mating and the introduction of sperm into the munization via the administration of an antibody, the vagina is caused by antibodies in the cervical mucus which do not disrupt the weight of the administered antibody - agglutination of the sperm in the cervix and usefully in a direct relationship to the prevent one further transport of the sperm into the leeches therapeutic effect. The titer of the antibody 20 uterus. The antibodies against the sperm or the biological effectiveness is the important factor. not only by agglutination, but also by inactivating the sperm from the point of view of dosing for passive immunization processes.

tion. The titer of an antibody relates to the maximum dilution of the antibody to formulate a contraceptive, the microparticles according to the invention containing a test situation can each have an effect. Two different contraceptives have also been used, which, in particular

Preparations of an antibody not based on the weight particular in women or in mammals, the desired amount comparable because it has a different biological contraceptive effect.

Have effectiveness. For each antibody to be administered by an example of a suitable contraceptive according to the invention, spermicidal compounds such as nonylphenoxypolyoxyethy-

an immunological titer of 1: 500 represents the minimum bio-30-ethenol, benzethonium chloride, (benzyldimethyl - [(2-

logical effectiveness. In addition, the Gesch- (l, l, 3,3-tetramethylbutylphenoxy) ethoxy) ethyl] -ammo-

with which the immunoglobulin or the antibody nium chloride), chloroindanol (7-chloro-4-indanol) etc. and the cervix, uterus and fallopian tubes natural and synthetic honnones such as progesterone (A4-

0.1 mg of the antibody per day not pregnen-3,20-dione), estradiol (estradioI-3,17ß-dicypionate),

exceed. Any dosage rate that is below 35 norethindrone (17-hydroxy-19-nor-17a-pregn-4-en-20-in-3-

this value and in relation to causing an on), norgestrel (d, l-13ß-ethyl-17a-ethynyl-17ß-hydroxy-4-

therapeutic response is effective is justifiable. gonen-3-one), ethynodiol diacetate (3 (3,17ß-diacetoxy-17a-

The antigen, antibody or contraceptive äthinyl-4-estreh), Lynestrenol (17a-Äthinylöstr-4-en-17ß-ol),

Microparticles containing agents can in suitable medroxyprogesterone acetate) 17ß-Hydroxy-6a-methylpregn-

Way by a known method for producing 40 4-en-3,20-dione), dimethisterone (17β-hydroxy-6a-methyl-17-

drug-containing microparticles. L-propinyl-androst-4-en-3-one), megestrol acetate (17a-Hy-

is the amount of antigen or antibody and droxy-6-methyl-pregna-4,6-diene-3,20-dione acetate), chlorine:

hormone regulating the menstrual cycle if a dinone acetate (6-chloro-17-hydroxy-pregna-4,6-diene-3,20-

such should be present, not critical, but dione acetate), ethynyl estradiol (17a-ethynyl-l, 3,5 (10) -oestra-

the microparticles normally contained about 10 to 60% by weight, 45 triene-3ß-diol), mestranol (3-methoxy-19-nor-17a-pregna-

preferably 10 to 50% by weight and in particular 10 to 25 l, 3,5 (10) -trien-20-in-17-ol) etc. Another group of

% By weight of the antibody or antigen. Compounds which are suitable for the micropart

In clinical medicine, the primary limitation will be used to cause premature abortion in women or to generate passive immunization in a mammal. Examples of suitable ones

Patients by means of administration of antibodies thereby 50 compounds that have an abortive effect are anti

stipulated that antibodies produced in animals histamines, cytotoxins, ergot alkaloids, hormones,

are relatively often a serum disease or anaphylaxis. Prostaglandins such as the prostaglandind Ez and F2 (1 la,

Causes when injected into humans as recipients 15 (S) -dihydröxy-9-ketoprosta-5-cis-13-transdienoic acid or

the. According to the invention, however, this problem is solved by 9 a, 11 a-15 (S) -trihydroxy-prosta-5-cis-13-trans-dienoic acid,

bypassing the local supply of microparticles because not 55 sympatholytic compounds, etc. Of course, only smaller doses of the antibodies are required, but also mixtures of different contraceptives or systemic administration of antibodies means in which the individual compounds are used.

is avoided. are biologically compatible.

In some cases, e.g. B. for permanent protection against the amount of microparticles inserted into the vagina

Infectious diseases, active immunization is more dependent on the amount of contraceptive than passive immunization. In the case of an antigen to the uterus and the fallopian tubes, which must be fed to the uterus and the fallopian tubes, the microparticles according to the invention, in order to achieve the desired contraceptive effect, will therefore produce antibodies. In the case of hormones excreted by a contraceptive, which not only produce the desired immune effect, the dose of the biological effect to be administered, but in comparison with the 65 hormone, is in the range from 20 to 1000 mg per day. In the event of

The type of antibody that is responsive to a systemic of the spermicidal contraceptive agent is at the

Immunization are produced even structurally and not necessary for most days of the menstrual cycle

are fundamentally unique. Systemic antibodies will be given a daily dose. It's only on some days

around the middle of the cycle, if conception is possible, it is necessary to administer a spermicidal agent at a dose level sufficient for contraception. In general, a daily dose of 25 to 1000 ng over a period of 7 days, which is approximately in the middle of the cycle, leads to the desired effect. Other types of contraceptives that can be used, as set forth above, include abortives. The administration of these abortives should immediately follow an omitted menstrual period, for 3 to 5 days at a dose of 1 to 500 mg per day.

All known methods for forming microparticles can be used in the production of microparticles according to the invention. The choice of a particular method depends primarily on the technical requirements of the matrix material and on the particular way in which the microparticles are to be used. In general, the microencapsulation processes can be classified into three basic types: (1) phase separation processes, e.g. Process for the separation of aqueous and organic phases, melt dispersion and spray drying; (2) reactions at interfaces, e.g. interfacial polymerization, in situ polymerization and chemical vapor deposition, and (3) physical processes. e.g. the fluidized bed spray coating, the centrifugal coating with one and more nozzles or openings, the electrostatic coating and the physical separation from the vapor phase.

The microparticles of the invention can be introduced into the vagina by a variety of methods. Preferably, a fixed number of microparticles are placed in a container intended for easy manual insertion into the vagina. The introductory container should be made from a biodegradable material that dissolves within minutes of being inserted into the vagina, releasing the microparticles. Pharmaceutical type gelatin capsules can suitably be used as a microparticle delivery device. The dose level can be changed by increasing or decreasing the number of microparticles in the feed device. Of course, other methods or this preferred method can be used as possible variations. For example, the microparticles can be shaped to form a solid vaginal suppository that offers the simplest and most direct method of using the microparticles by using a suitable suspension medium such as gelatin. Creams, jellies, foams or liquids can be used as a suspension medium for microparticles. Preparations of this type can be made using a refillable syringe or certain pressurized vaginal delivery devices, e.g. an aerosol device, or tubes or flasks to be squeezed into the vagina. A variety of different types of delivery devices intended for the introduction of pharmaceutical agents into the vagina and rectum are commonly used. A gelatin capsule is also a suitable carrier for the supply of microparticles.

The invention is illustrated by the examples below.

Example 1 Preparation of progesterone-containing polylactic acid microcapsules

2.5 g of progesterone and 10.0 g of d-1 polylactic acid were added

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dissolved in 38 g of methylene chloride. The resulting viscous solution was poured into a 250 ml kettle containing 120 ml of a 5% by weight aqueous polyvinyl alcohol solution. The dispersion obtained was stirred at about 200 rpm until a stable emulsion had formed, the droplets of which had a diameter in the range from 50 to 100 μm. A vacuum was applied to the emulsion until it started to foam, whereupon the stirring speed was reduced to 600 rpm. After 2 h most of the methylene chloride had evaporated. Furthermore, continuous stirring was not required to prevent the microcapsules under development from agglomerating. The emulsion was then centrifuged, the aqueous polyvinyl alcohol solution was decanted, and the microcapsules were resuspended in 150 ml of deionized water. A vacuum was then continuously applied to the stirred, aqueous suspension over a period of about 18 h. The suspension was then centrifuged and the microcapsules obtained were washed with water and then collected by vacuum filtration. The microcapsules were dried at room temperature overnight under a strong vacuum and then sieved to give a fraction between 43 and 61 (im. This procedure gave microcapsules containing 22 + 1.5% by weight progesterone.

Example 2 Preparation of progesterone-containing glycerol monostearate microcapsules

1.0 g 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. For cooling, 601 / min nitrogen was allowed to flow into the device while the flow of nitrogen introduced into the sprayer to aerosolize the molten mixture was adjusted to the maximum speed of 5,751 / min. The aerosol was sprayed intermittently and the microcapsules were collected and sieved to collect a size fraction between 43 and 61 µm. The microcapsules made by this method were spherical and contained a theoretical fill of 20% by weight progesterone.

test

Four female baboons were injected with d, l-polylactic acid microcapsules containing 7.87 mg of progesterone, while five female baboons were treated by vaginal administration with d, l-polylactic acid microcapsules containing 7.87 mg of progesterone. All baboons were treated on day 5 and uterine biopsies were taken on day 12. The daily blood levels of estradiol and progesterone were also measured. The results of the daily determinations are shown in Fig. 11, where Fig. 11A shows the progesterone and estradiol levels for intramuscular injection of the microcapsules, while Fig. IIB shows the progesterone and estradiol levels for vaginal administration of the microcapsules. It is evident that the baboons treated by intramuscular injection found a higher blood level of progesterone than the baboons treated vaginally. There are two ways to explain this fact: (1) Not all microcapsules that have been inserted into the vagina remain in the body over the 7-day treatment period; (2) The progesterone that is released from the vagina after the microcapsules have been brought into the vagina reaches the bloodstream

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not completely. These options are clearly visible (see Fig. 13b). Due to the continuous both lent a contribution to the differences in systemic treatment with progesterone between the 5th and 12th see progesterone levels between the two treatment days, the formation of numerous protrusions is grouped. When the systemic administration of progesterone causes form of microvilli. Looking at the epithelial surface alone and the possibility of local, direct feeding of baboons treated by injection, delivery to the uterus in the intravaginally treated microvilli occur in an even distribution while excluding animals, it is logical, at Baboons, they in baboons that were treated intravaginally, in the form that were treated by injection, have a more intense effect, which is genetically separated from individual or separate, spotty areas than in vaginally treated baboons. The patchy, uneven distribution is waiting. The results of dose-response experiments from Microvilli occur in the form of areas where these support this expectation. However, this assumption will be lacking in epithelial cells of microvilli, and other areas in which a comparative histological examination of the biopsies shows that the size of the microvilli varies. of the baboons' endometrium. The histological- This uneven distribution of a progesterone test does not show a significant reduction in the induced change in the morphology of the upper level of the progesterone-induced secretory surface epithelium indicates localized areas of stimulation activity in the case of intravaginal administration- treated by progesterone. Following a systemic Zu-Baboon in comparison with Baboons that received intramus guidance, all areas of the endometrium had been treated with a kular injection. This is in the even dose of the drug, while when looking locally at the difference in systemic progesterone delivery certain areas of higher or lower gel are somewhat unexpected. Indeed, through this, 20 doses can be obtained, depending on where the micron-end contradicting findings locate the possibility of local capsules within the uterus. Delivery of progesterone to the vaginally treated patients. Figures 14A and 14B confirm the middle progestin vian. Figures 12a and 12b show the histologinon level and the mean estradiol level of baboons see the appearance of the endometrium of a baboon female, compared either by intramuscular injection by intravaginal administration of microcapsules 25 (Figure 14A) or by intravaginal Administration containing 7.87 mg of progesterone (with H (Fig. 14B) under the conditions described above, 4-E stained tissue at 20 or 100 times magnification - with a dose of 1.57 mg of progesterone containing Mi. -rung). 12c and 12d show the same tissue (which had been treated with krokapsetn. This low dose of PAS stained) at 20 and 100 times magnification, respectively. of microcapsules has a slight inhibition

A second route to histological evidence, which has the effect on estradiol level when administered by ingestion of progesterone in the vaginal treatment injection, and has no effect when confirmed in animals the observation that is administered intravaginally. In baboons treated for injections caused by progesterone, exogenous progesterone is between endometrial stology in baboons treated with low blood levels by infection on the 5th and 12th days. present in the entire endometrium. However, in baboons treated vaginally, while the effects in baboons that were treated could be localized and within the sensitivity range treated by intravaginal introduction of the microcapsules ranging from the 5th to the 12th analysis method no exogenous progesterone can be found in the blood from gland to gland for various days, the intensity of the stimulation in the surface. On the basis of the comparative glands shown in FIG. 14, which lie below the surface epithelium, 40 details are striking the hormones and when excluding (Fig. 12). The absence of subnuclear vacuoles in the possibility of local delivery, it seems logical, to the surface glands in pa treated by injection, that the histology of the endometrium of the vaginal vianas and the large amount of subnuclear vacuoles, baboons treated in normal, did not control resembles the pattern observed in the surface glands of intravaginally treated animals. The logi baboons appear to prove the local intrauterine supply. The basis for this expectation lies in the fact that progesterone levels in the intravaginally treated Pa progesterone and estradiol blood levels in baboons, the vianas. have been treated intravaginally with the low dose of micro-A comparative study of the morphology of the capsule, which is identical to the blood level normally seen on surface epithelium by scanning electron microscopy. Therefore, based on further evidence for localized gestagenic Ef- the endocrine data there is little reason to expect fects following the intravaginal treatment with the hormone-induced changes in the hi-microcapsules. In Fig. 13 the morphology of the uterine endology of the endometrium. For animals compared by injecting nasal surface. It shows: have been treated, but the same

13a clarify the normal baboon endometrium on the 12th day, because between the 5th and the 12th day in the arrival

without treatment; following the treatment of a detectable progesterone level

13b the normal endometrium was found on the 20th day without gel in the blood. In addition, the Östra-

Treatment; diol blood levels compared to normal estradiol

13c the endometrium on the 12th day in the case of exposure in animals not treated as a control to be somewhat reduced by 7.87 mg Pro intramuscular injection. Again the histological

microcapsules containing gesteron and 60 finds contradicting the expected results. In spite of

13d shows the endometrium on the 12th day in the absence of measurable, systemic progesterone when treated by intravaginal introduction of 7.87 mg of prop baboons, which were treated by vaginal route, shows microcapsules containing sterone. the endometrium has clear gestagen effects.

Progesterone calls for the formation of pronounced pre-examining the morphology of the surface epithelium

Cracks in the form of microvilli on the luminal surface 65 by scanning electron microscopy shows the frequent

surface of the glandular epithelial cells. Before ovulation occur from microvilli, which is clear evidence of the stimulation

(see Fig. 13a) are normally not shown to be micro-ionized by progesterone (see Fig. 15). The villi are present; however, after ovulation, they are relatively Figs. 15a and 15b are photomicrographs of the surface

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Chen epithelium of the treated baboon on the 12th day of the menstrual cycle with low (9600) and high (18 240) enlargement. Figure 15c is a photomicrograph of the epithelium showing microcapsules containing progesterone on the surface of the epithelium, while Figure 15d is a microphotograph of the microcapsules themselves. Although the changes in the histology of the endometrium caused by progesterone could occur in response to systemic progesterone, which could not be measured due to its low concentration by the analytical system used, it is a more likely explanation that the changes caused by progesterone are a result the direct, local, intrauterine supply of progesterone.

The most important point to emphasize is that with a low dose of microcapsules (i.e. with 7.87 mg of progesterone) secretory changes were caused in the endometrium. In addition, a response to the dose associated with the changes was shown, suggesting that it should be possible to achieve an effect sufficient to inhibit reproductive function by increasing the dose.

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16A and 16B, the pattern of contractile activity of the cervical muscle of two baboon females on the same day of the menstrual cycle is compared. A special sensor or transducer that was built as it was by W.D. Blair and L.R. Beck in Ovum Transportand Fertility Regulation, 1976, pp. 41-74, Scriptor Publication (Copenhagen) was introduced into the cervix of each baboon female and used to measure the diagram of the cervical contractions shown in the figures. The sensor or transducer was connected to a chart recorder. Figure 16A shows the results obtained on a baboon female which had been treated by inserting estradiol-containing microparticles into the vagina, while the results obtained on a baboon female which was not treated with microparticles for control are shown in Figure 16B . The results show that the treatment stimulates both the frequency and the amplitude of the cervical contractions. These contractions move the microparticles through the system.

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8 sheets of drawings

Claims (10)

645 545 2nd PATENT CLAIMS 1. microparticles for the treatment of internal female genital organs, characterized in that they (a) an antigen or an antibody and a menstrual regulating hormone or (b) contain a contraceptive and a menstrual regulating hormone incorporated into at least one biologically compatible and biodegradable matrix material, the microparticles having been introduced into the vagina by the natural mechanism of transport of the female internal genitalia through the cervix into the Uterus and fallopian tubes. 2. Microparticles according to claim 1 for the active or passive immunization of the internal female genital organs, characterized in that they contain an antigen or an antibody, which are incorporated into a biologically compatible and biodegradable matrix material, in an amount sufficient to provide an immune response The microparticles can be transported through the cervix into the uterus after their introduction into the vagina through the natural mechanism of the female internal sex organs. 3. Microparticles according to claim 1 or 2, characterized in that they have a size of 20 to 70 microns and contain as a matrix material a polymer or polycondensate of lactic acid or glycolic acid or a copolymer of glycolic acid and lactic acid. 4. Microparticles according to claim 2 or 3, characterized in that they contain 10 to 60 wt .-% of the antigen or antibody. 5. Microparticle according to one of claims 2 to 4, characterized in that the antigen is derived from a bacterial or viral pathogen and that the antibody responds to an antigen from a viral or bacterial pathogen. 6. Microparticles according to claim I, characterized in that they contain a hormone regulating the menstrual cycle and a contraceptive agent, which are incorporated into a matrix material, and that they have a sperm proxy effect, the contraceptive agent in the event that it is a contraceptive hormone, a hormone different from the menstrual cycle regulating hormone, and wherein the menstrual regulating hormone can stimulate the natural delivery mechanism after the microparticles have been inserted into the vagina. 7. Microparticles according to claim 6, characterized in that they have a size of 20-70 µm and that the biodegradable and biologically compatible matrix material has been selected from polymers or polycondensates of glycolic acid, polymers or polycondensates of lactic acid and mixtures thereof . 8. Microparticle according to claim 6 or 7, characterized in that the hormone regulating the menstrual cycle, which causes the transport, is an estrogen or a progestogen. 9. microparticles according to any one of claims 6-8, characterized in that the contraceptive agent selected from progesterone, estradiol, norethindrone, Norg-estrel, ethynodiol diacetate, lynestrenol, medroxyprogestone-ron acetate, dimethisterone, megestrol acetate, chloromadinona cadiol and ethynestrestrate Hormone is. 10. Agent for the treatment of the internal female genital organs, containing microparticles according to one of claims 1-5, characterized in that it is in the form of a suppository, a cream, a jelly, a foam or a liquid with a pharmaceutically acceptable binder or a pharmaceutical compatible shell is formulated. 10th