JPS61115429A - Biological method - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a method for increasing the fertility of immature sows, such as sows up to 5%.

Because of their economic importance, research in the reproductive biology of livestock has long been devoted to methods for manipulating the visual capacity of farm animals.

Ovulation rate in sheep can be increased by immunizing sheep against steroidal androgens or steroidal estrogens, for example with conjugates of steroidal androgens or steroidal estrogens and immunogenic proteins. It has been shown.

In adult sows, the limiting factor for fertility (ie production scale) is often uterine capacity rather than ovulation rate, as is the case in multiparous ewes. This is not always the case in immature sows such as gilts, ie nurse sows which have a lower ovulation rate fti than adult sows. For example, in nulliparous sows, the ovulation rate increases with increasing sexual age (number of estrus cycles) until about the fourth estrus period after puberty. The sow is #! In the 11th postpartum estrus, the ovulation rate may decrease due to the decreased nutritional status due to early lactation. This effect is not exhibited by pregnant sows. It has been found that in the case of immature sows, fertility can be increased by increasing the ovulation rate, and in fact it may be advantageous to increase the ovulation rate of adult sows.

Now 5 the production of sows, especially immature sows, by immunizing them against one or more of such steroids, for example using conjugates of steroidal androgens or steroidal estrogens and immunogenic carriers. It has been found that it is possible to increase the ovulation rate to more fully utilize the uterine capacity and ultimately their reproductive potential.

According to one aspect of the present invention, there is provided a method of increasing ovulation rate in a sow comprising immunizing the sow with one or more steroidal androgens or steroidal estrogens.

It will be appreciated that low ovulation rates in immature sows are a natural phenomenon and immunization according to the invention has the effect of increasing the natural ovulation rates of such animals to higher levels.

As used herein, the term steroidal androgen refers to all steroidal substances that stimulate the development of secondary sexual characteristics in males. Androgenic activity can be assessed by measuring the regeneration of the retracted crest of castrated male rats following androgen administration or by measuring the growth response of the seminal vesicles of castrated male rats following androgen administration. As used herein, the term steroidal estrogen refers to all steroidal substances that stimulate the development of secondary sexual characteristics in females. Nistromene activity can be assessed by measuring uterine growth or vaginal epithelial keratinization following estrogen administration to ovariectomized female rats or mice.

Immunogenic conjugates between an immunogenic carrier and a steroidal androgen derivative or a steroidal estrogen derivative for use in the method according to the invention can be prepared by techniques known in the art (for example in the British patent 5). Second,
No. 071,107 (ORISO)). Suitable steroidal androstene derivatives include, for example, androst-4-en-3,17-)one (androstenedione) or testosterone.

Suitable steroidal estrogen derivatives include, for example:
Derivatives of estrone are included.

Preferred immunogens are therefore immunogenic carriers, such as immunogenic proteins such as androst-4, which are functionalized with acid groups at the 1, 7, 11 or 15-positions of the steroid ring.
- Manufactured by bonding with a 3,17-dione fatty conductor.

To prepare immunogens against testosterone, testosterone derivatives modified with an acid group at the 3- or 17-position can be used.

The steroidal androgen derivative is thus testosterone-6-carboxy-methyloxime or preferably 11α-hydroxy-androst-4-ene-
3,17-dione-11α-hemisuccinate or 7
α-(2-carboxyethylthio)androst-4-
It can be oxidized with ene-6°17-dione. Preferred estrone conductors are those that generate estrone-specific antibodies after being conjugated to a protein and administered to an animal. Non-limiting examples of such estrone inducers already known in the art include estrone-6-hemisuccinate, estrone-5
-carboxymethyl ether, estrone-6-carboxy-methyloxime and 15-carbodimethylestrone. A preferred steroidal estrogen is U-estrone-3-carboxymethyl ether.

Immunogenic carriers will generally be high molecular weight proteins that are antigenic to animals. Certain naturally occurring antigenic proteins are of particular interest, such as homologous and heterologous serum albumin and thyroglobulin. Generally human serum albumin is preferred.

Generally, the % immunogenic composition may additionally contain an adjuvant, diethylamine ethyl dextrin,
Renhui's excessive adjuvant, which is known in the art industry, may be used.

One aspect of the invention is administering to a sow an immunogenic composition comprising at least one immunogenic conjugate between an immunogenic carrier and an androgenic steroid derivative or an estrogenic steroid derivative. The present invention relates to a method for increasing the ovulation rate of sows, particularly immature sows, comprising:

Another aspect of the invention is a sow in a herd.

In particular, it relates to a method of increasing ovulation rates in a herd of sows, which comprises immunizing immature sows with one or more steroidal androgens or steroidal estrogens.

According to another aspect of the invention, a sow is immunized with one or more steroidal androgens or steroidal estrogens prior to the start of the mating season, thereby producing steroid-binding antibodies in the animal. This is because the ovulation rate is present during the mating period at a titer high enough to significantly increase the rate of ovulation, but low enough to ensure that virtually all animals exhibit oestrus before the end of the nine mating periods. A method for increasing the fertility of sows, particularly immature sows, is provided.

Therefore, for example, steroid-based ANDC! The average steroid-binding antibody titer at the beginning of the mating season is 1/10 to 115,000, preferably 1/30 to 115 after being immunized against one or more steroidal estrogens.
000 and this titer will be 1/1 at the end of the period
0 to 1/10000 preferably 1/60 to 1/1000
It is desirable that immunity be increased, especially in immature sows such as gilts, by maintaining the immunization range in the 0 range.

Steroid antibody titers in the case of immunotherapy with conjugates between an immunogenic carrier and eg androstenedione can be measured using a dextran plate & charcoal radioimmunoassay procedure. Serial dilutions of plasma were prepared in slow motion and 600 samples were collected (1, 2, 3, 7-
5H) Androstenedione (@min 15000 disintegration =
20-25 pg) 100μβ and incubate overnight at 4°C. After bed warming, the bound 7-lactone and Yuto's 72-kusion were separated using dextran-coated charcoal cloud M (500 μβ) and the radioactivity in the bound fraction remaining in the supernatant was determined. The androstenedione antibody titer of the sample is a dilution of plasma that binds 40% of the total radioactivity added.

It is desirable that the animals to be immunized be of age and in moderate to good condition at the time of mating.

Generally, the initial dose of hormone conjugate for immunization of #monobutsu will be 0.6-51 Jljl. Therefore, for example, a gilt sow weighing 100# has a zygote of 12g9~5.6'a.
9. Preferably, 2.411p can be accepted first.

K to achieve the steroid-binding antibody titers described above.
ri One or more booster doses at appropriate intervals, such as 14 to 65 days, preferably 21 days, may be desirable. Thus, for example, the total effective amount of hormone conjugate can range from 0.6 to 8.0'M9. Administration of the compositions is most conveniently carried out parenterally, eg subcutaneously, subcutaneously or intramuscularly.

Generally, the amount of composition given at each administration is 1-6 ml;
The concentration of hormone conjugate in the composition, which will preferably be in the range of 4 ml, will generally be between 0.2 and 5119/ml.

Any immunization technique can be used to obtain the desired anti-steroid antibody potency.

The invention is illustrated by the following non-limiting examples.

Example (a) Steroid-Protein Immunogen Composition allI
7α-(2-carboxyethylthio)androst-4-en-3,17-dione (5-) in Nll
A solution of isobutyl chloroformate (28
It was treated with a solution of ■. This solution was kept at approximately 10°C (275°C) for 40 minutes with occasional shaking. This was then mixed with human serum albumin (H8A) (275cm) in 0.1M phosphate buffer (10°C) at 7.8°C. 2 in the stirring solution
Added once for ~3 minutes.

After 24 hours, the slightly cloudy solution was diluted with distilled water (500
Dialysis was performed at 5° C. for 192 hours (replaced 11 times). The retentate was lyophilized to obtain a steroid-protein-* threaded complex as a white foam (194 cm). Its UV absorption at 246 nm corresponded to 26 mol of steroid uptake.

(bl, z, steroid-protein immunogen-azidipant composition, the steroid-protein conjugate (12■) manufactured by J4) was added to 0.9
A paste was made in sterile salt water (0.2 fflt). This host was further added with 0.9% sterile saline,
Gradually dilute with gentle stirring to make total itlOm.
Q. On the other hand, DKAJ! i-dextran 1
Dissolve 5jl in distilled water (100ml) and add Tris-
By constantly adding (dimethyl in hydro)-methylamine! By adjusting the pH to 7.5-Z7 and then diluting to 150 μm with distilled water! l)
A stock solution of DEAR-dextran was prepared. Immunogen
The adipant composition was made by adding 10 ml of DEAE-dextane stock solution to 10 ml of the conjugate solution in saline.

(C) Immunization and death of nulliparous sows for 150-170 days and immunization with steroid-protein immunogen-azidipant composition as 4 ml of steroid-protein immunogen-azidipant composition at 14-day intervals. -Dextran 2
Androstenedione in Tonaka - Human serum albumin T2
An amount equivalent to q was injected. Blood samples were taken at intervals to determine if the pigs developed any antibodies against androstenedione. The results showed that antibody formation was successful in 100% of the treated animals and the titers produced ranged from 1/10 to 1155.
It showed that it was 00.

Control animals remained untreated.

The mean period of estrus (SJI!iM) found for the first time was 263±5 days in controls and 246±7 days in non-injected pigs.

Each sow that has been treated with
Received ~7 injections.

During the tail phase, the arithmetic mean (84!fM) antibody titer against androstenedione decreased to 1/6 in 230 to 239 days.
26±194 to 11544±41 in 250-259 days
The range was up to 3.

(d) Production scale In production scale data, immunized gilts were compared to control (91
±0.5, n=8 siblings) is also quite large (P(0,0
5) Siblings (10,7 ± 0.7. Mean ± 81!, M,
n=11 siblings).

Patent applicant: 2 people outside of Gracs Group Limited

Claims (1)

Claims: 1) A method of increasing ovulation rate in sows comprising immunizing the sow with one or more steroidal androgens or steroidal estrogens. 2) A method of increasing the ovulation rate of a herd of sows comprising immunizing the sows in the herd to one or more steroidal androgens or steroidal estrogens. 3) Immunization of sows with one or more steroidal androgens or steroidal estrogens prior to the start of the mating season, whereby steroid-binding antibodies in the animals significantly increase the ovulation rate during the mating period. A method for increasing the fertility of sows, comprising: being present at a titer high enough to produce fertility, but low enough to ensure that substantially all animals exhibit oestrus before the end of said mating season. . 4) The average steroid-binding antibody titer is between 1/10 and 1/5,000 at the beginning of the mating season and between 1/10 and 1/10,000 at the end of the mating season. The method described in Section 3. 5) administering to a sow an immunogenic composition comprising at least one immunogenic conjugate between an immunogenic carrier and an androgenic steroid derivative or an estrogenic steroid derivative; How to increase ovulation rate. 6) The method according to any one of claims 1 to 5, wherein immunization of immature sows or sows is carried out. 7) A method according to any one of the preceding claims, wherein the immunization is carried out against androst-4-ene-3,17-dione and/or testosterone or estrone. 8) Androst-4-ene-3,1 functionalized with an acid group at the 1,7,11 or 15-position of the steroid ring
8. A method according to any one of the preceding claims, wherein the immunization is carried out by administering an immunogenic conjugate between a 7-dione derivative and an immunogenic carrier. 9) Androst-4-ene-3,17-dione derivative is 11α-hydroxy-androst-4-ene-3,
17-dione or 7α-(2-carboxyethylthio)-androst-4-ene-3,17-dione. 10) The immunization is carried out by administering an immunogenic conjugate between a testosterone derivative functionalized with an acid group in the 3- or 17-position and an immunogenic carrier. The method described in any one of paragraphs. 11) Claim 1, wherein the testosterone derivative is testosterone-3-carboxymethyloxime.
The method described in item 0. 12) Any one of the preceding claims, wherein the immunization is carried out by administering an immunogenic conjugate between an estrone derivative functionalized in the 3-position and an immunogenic carrier. The method described in section. 13) The method according to claim 12, wherein the estrone derivative is estrone-3-carboxymethyl ether. 14) A method according to any one of the preceding claims 8 to 13, wherein human serum albumin is the immunogenic carrier. 15) A method according to any one of the preceding claims, wherein an immunogenic composition containing an adjuvant is used. 16) The method according to claim 15, wherein the adjuvant is diethylaminoethyldextran. 17) Claims 1, 2, and 2 consisting of an immunogenic conjugate between an immunogenic carrier and a steroidal androgen derivative or a steroidal estrogen derivative.
An immunogenic composition for use in the method according to item 3 or 5. 18) Between an immunogenic carrier and a steroidal androgen derivative or a steroidal estrogen derivative for the preparation of an immunogenic composition for use in the method according to claim 1, 2, 3 or 5. Use of immunogenic conjugates.