JP2000508293A - Novel peptide useful for promoting FSH action - Google Patents

Abstract

  (57) [Summary] Novel peptides useful for promoting the activity of follicle stimulating hormone (FSH) and their use in enhancing fertility are provided.

Description

DETAILED DESCRIPTION OF THE INVENTION                 Novel peptide useful for promoting FSH action   The present invention is useful for promoting follicle stimulating hormone (FSH) bioactivity in mammals. A novel peptide for use. In particular, the present invention relates to such peptides And especially humans, sheep, cows, pigs, goats, cats, dogs, horses, camels, llamas And their use in stimulating FSH activity in alpaca. You.   Follicle stimulating hormone (FSH) is luteinizing hormone (LH), thyroid stimulating hormone. Glycoproteins such as mon (TSH) and chorionic gonadotropin (CG) Belongs to the group of These are all two different non-shared, termed α and β Consists of binding subunits. In some species, the different hormones The α subunits are identical in amino acid sequence. Β-subunit specific to hormones Knits show different amino acid sequences (Comment, Combarnous Endocrine Reviews, 13: 670-691 (19 1992)).   FSH plays a key role in regulating folliculogenesis and spermatogenesis Fulfill. The presence of FSH indicates the recruitment of quiescent follicles and the antral follicle Is essential for its development into humans (for review, Hillier, Human See Production (Human Reproduction), 9: 188-191 (1994). ). These biological effects are due to the binding of FSH to specific cellular receptors. Such receptors are referred to as receptor recognition sites, FSH Binds to specific sites on the molecule (Combarnous, supra). different Animal Using Dell, the effect of FSH on follicle growth and ovulation was studied. These For hypophysectomized mice (Wang, Greenwald, Endocrinology, 132: 2009-2016 (1993) ); Wang and Greenwald, J. Reprod. Fert., 99: 403-413 (1993)) and sexual function Depressed mice (Halpin and Charlton, Jaeplodo   J. Reprod. Fert., 82: 393-400 (1988); Katana. Cattanach et al., Nature, 269: 338-340 (1977). Mason et al., Science, 234: 1366-1371. (1986)). In particular, Snell and Ames dwarf mice (Snell a nd Amesd warf mouse), FSH supplementation showed total healthy follicle counts in 24 day old animals And the ability to increase uterine and ovarian weight (de Reviers ers), Acta. Endocrinologica, 106: 121. -126 (1984)).   The effects of antibodies on FSH bioactivity were also studied using animal models. antibody The effect of is usually considered with respect to the neutralization of biological activity. However, monochrome It has been found that a local antibody (MAb) can also improve FSH activity (Glenk Ross, Glencross et al., Journal of Endoclinology rinology), 136: R5-R7 (1993)). In the above study, Monoclonal antibodies prepared against FSH (bFSH) To promote the ability to induce uterine growth in Snell dwarf mice Was. However, long-term administration of mouse MAbs to other species has deleterious consequences. Is brought.   Thus, administration of a selected peptide sequence of FSH from the appropriate species will It is preferred to obtain a site-specific FSH-promoting antiserum. One of the subunits and the other In common with glycoproteins, it is produced against whole hormones or whole hormones. Effect of other antibodies on the bioactivity of other glycoprotein hormones Problems may arise with regard to. Therefore, the commonality of the structure with FSH is shared. Has little effect on the biological activity of other glycoprotein hormones Produce antibodies that are specific or have no stimulation of FSH activity Is the purpose.   The present inventors have now found a range of peptides that achieve the above results. these Are the amino acid residues 33-47 and 40-51 of the FSH molecule (See FIG. 1). This region is included as a receptor recognition site The above is itself surprising. Therefore, the FSH The binding of the antibody must not be inconsistent with the binding of FSH to its receptor (Santa Santa Coloma et al., Biochemistry, 29: 1194. ~ 1200 (1990); Santa Coloma and Lightchart (R) eichert), J. Biol. Chem., 265: 5037-50. 42 (1990); Lapthorn et al., Nature, 369: 455-461 (1994)). The peptide induces the promotion of FSH bioactivity. For the range of mammalian species in which it occurs, it may be administered alone or in combination. Good.   Therefore, according to the first concept, the present invention provides a peptide having the following sequence: What to offer:   (I) YTRDLVYRDPARPNI;   (Ii) YTRDLVYKDPARPNI;   (Iii) YTRDLVYKDPARPKI;   (Iv) RDPARPNIQKTC;   (V) KDPARPKIQKTC;   (Vi) KDPARPNIQKAC; or   (Vii) KDPARPNIQKTC, Or a sequence substantially homologous thereto.   The peptides of the present invention may be those wherein the antibody enhances either endogenous or exogenous FSH activity. Is useful to elicit an antibody response in a mammal that causes This further Then, ovulation or spermatogenesis, ie, promotion of fertility, occurs.   The one or more peptides may be one that recognizes FSH in the test mammal. It can be provided as a vaccine used to stimulate an epidemic response. Wakuchi like this Form the second concept of the invention.   According to a third concept, the present invention provides a nucleic acid sequence encoding the peptide, or It provides a sequence that is substantially homologous to this. Such a nucleic acid sequence May be a DNA sequence or an RNA sequence. Such a nucleic acid sequence Encodes the amino acid of interest due to the degeneracy of the genetic code, or And all other nucleic acid sequences having substantial homology to such sequences.   In the present specification, the substantial homology at the amino acid level means a significant number. That the peptide sequence having the constituent amino acids has homology to the above peptide sequence Means For example, at least 40%, 50%, 60%, 70%, 80%, 9 0%, 95% or 99% (larger is preferred) has homology.   Alternatively, the peptides of the invention can be used to produce antibodies, and these antibodies are Further to elicit enhanced bioactivity of either endogenous or exogenous FSH Administered to mammals. Therefore, to further concepts Thus, the present invention provides antibodies having specificity for one or more of the peptides of the present invention. To offer. In particular, said antibodies are monoclonal antibodies.   According to one embodiment of the above concept of the present invention, the antibody comprises one or more peptides of the present invention. Formed as an antibody preparation, consisting of one or more antibodies reactive with the tide.   According to a preferred embodiment of the above concept of the present invention, peptide (i) or (iv) ) Promotes sheep FSH activity or promotes sheep FSH activity Peptide (i) is used to raise antibodies for bovine or porcine For promoting FSH activity in bovine or porcine or pig. Peptide (i) is used to raise antibodies to horse or human F For promoting SH activity or for promoting FSH activity in horses or humans Used to raise antibodies; Peptide (ii) can be obtained from sheep or pig F To promote SH activity or to promote FSH activity in sheep or pigs Peptide (ii) may be used to generate horse, human or To promote FSH activity in horses, or in horses, humans or cattle. Used to raise antibodies for the development of peptides; Or to promote FSH activity in humans or to promote FSH activity in horses or humans. Used to raise antibodies for transit; peptide (iii) is a bovine, For promoting FSH activity in sheep or pigs, or in cattle, sheep or pigs Used to raise antibodies to enhance FSH activity of the peptide (i v) promotes bovine FSH activity or promotes bovine FSH activity Used to raise antibodies for porcine, equine or Promotes FSH activity in humans, or in pigs, horses or Used to raise antibodies to enhance FSH activity in humans; Peptides (V) promoting horse or human FSH activity, or Used to raise antibodies to enhance FSH activity of the peptide; ) Is for promoting FSH activity in cattle, sheep or pigs, or Used to raise antibodies to enhance di- or porcine FSH activity; Peptide (vi) promotes sheep FSH activity or sheep FS Used to raise antibodies to enhance H activity; peptide (vi) To promote FSH activity in cattle, cows, pigs, humans or horses, or in cattle, pigs Used to raise antibodies to promote human or horse FSH activity Peptide (vii) to promote porcine FSH activity or porcine FSH activity. Used to raise antibodies to enhance SH activity; peptides (vii ) Is for promoting FSH activity in cattle, sheep, humans or horses, or Producing antibodies to enhance FSH activity in sheep, humans or horses used.   The peptide or antibody of the present invention is preferably in the form of a pharmaceutically acceptable formulation Is administered. Thus, according to a fifth concept, the invention relates to one or more of the inventions A peptide, or one or more antibodies or antibody preparations of the invention, and one or more antibodies. Providing a pharmaceutical formulation comprising a pharmaceutically acceptable carrier, diluent or excipient. is there.   According to a sixth concept, the present invention relates to a method for promoting FSH bioactivity in a mammal. It provides the use of the peptides or antibodies of the invention. According to a specific embodiment And bioactivity is cattle, sheep, horse, pig, human, camel, goat, cat, dog , Llama or alpaca.   According to a seventh concept, the present invention relates to a method according to the invention, comprising one or more peptides or one or more peptides. Administering the antibody to a mammal, particularly a mammal. Rhino, sheep, horse, pig, human, camel, goat, cat, dog, llama or alpa It is intended to provide a method for promoting FSH bioactivity in mosquitoes.   The present invention will be described with reference to the following examples. There is no restriction.   The examples refer to the following drawings:   FIG. 1 shows a comparison of the sequences of the β subunits of FSH from various species. The   FIG. 2 shows in pre-immune serum (bleed 0) and also in sheep A1 (2-a ) And A2 (2-b) peptide A (bleed 4) and sheep B1 (2-b) c) whole bovine FSH (bFSH) in the antiserum after four doses of peptide B Shows a comparison between antibody titers against   FIG. 3 shows 3.3, 10, 30, and 90 μg / day of sheep FSH (oFSH) and Or Snell dwarf mice (Sne) given phosphate buffer (hormone diluent, PBS) uterus (3-a) and ovary (3-b) weight and keratin of ll dwarf mouse) group The average (± SEM) of the chemical conversion index (3-c);   FIG. 4 shows 20 μg / day oFSH (FSH group), 20 μg / day oFSH + Peptide A antiserum (group FSH + A), 20 μg / day oFSH + pre-immune serum (FS H + P group), peptide A antiserum alone (group A), and PBS control (group C) ) And ovary (4-a) and ( 4-B) weight and oF by measuring vaginal smear keratinization index (4-c) This shows the evaluation of antibody-mediated promotion of SH bioactivity; all values are average. Average ± SEM; and   FIG. 5 shows 20 μg / day oFSH (FSH group) and 20 μg / day oFSH. SH + peptide B antiserum (group FSH + B), 20 μg / day oFSH + preimmune blood Qing (FSH + P group), peptide B antiserum alone (group B), and PBS control Uterus (5a) and ovary (5b) ) And FSH physiological activity by measurement of keratinization index (5c) of vaginal smear FIG. 9 shows the evaluation of mediated antibody-mediated facilitation; all values are mean ± SEM It is.Example 1 Preparation of antiserum against peptide sequence of bFSH   Peptide: A peptide having a different FSH sequence was prepared by using an upgraded 431A ) Automatic peptide synthesizer (Applied Biosystems) F used_mocSynthesized by Fmoc chemistry. The peptide is free amino and It was synthesized to have a carboxyl end. The identity of the peptides was determined by MALDI (Ma Trick assisted laser desorption ionization (matrix assi sted laser desorption ionisation)) Analysis by reverse phase chromatography for precipitation showed a purity of greater than 95%. Was decided.   KLH conjugation: 20 ml of 10 mM sodium phosphate buffer (p H6.8) with 5 mg of peptide and 5 mg of keyhole limpet hemosine An (KLH) was added to the mixture; this solution was added to 50 μl of 25% glue. The mixture was stirred while tartaraldehyde was slowly added thereto, and stirred for 3 hours. Then this Divide the solution into 20 x 2 ml aliquots and store at 20 ° C for easy use. I kept it.   Animals and treatments: 10 mature ewes were divided into 5 groups. Two sheep, pea Peptides (i) and (iv) (referred to as A and B respectively in this study), and Each peptide was immunized:     HADSLYTYPBAT (denoted as C; 69th to 80th bFSH Array);     HCSKCDSDSTDC (denoted as D; bFSH sequence at positions 83 to 94); and     WCAGYCYTRDLVY (denoted as E; bFSH sequence at positions 27 to 39) .   Sheep were identified according to the immunized peptide (ie, peptide A = human Azaleas A1 and A2). The peptide / KLH complex (con jugate) in 2 ml complete Freund's adjuvant, One ml was injected into each hind limb of each sheep. Using incomplete Freund's adjuvant Three boosts were performed at three week intervals.   Radioimmunoassay: Serum samples were prepared before the first peptide injection (immunization). Before) and 10 days after each immunization was taken from each sheep. Each serum Liquid radioimmunoassay for a series of dilutions of the sample (1/10 to 1/6250) The assay tested the ability to recognize iodinated bFSH. Briefly, 50 μl of blood Wash the cells with 100 μl radioimmunoassay buffer (containing 0.05% Tween 0.05 M phosphate buffer) +50 μl iodinated bFSH (¹²⁵I; USDA b FSH-I-2AFP 5318C; 20,000 CPM / tube, saratin Salacinski et al., Analyt. Biochem., 117: 1. 36-146 (1981) using iodogen. ). After incubating the assay tubes at 4 ° C. overnight, 25 % Polyethylene glycol 6000 and 0.15% bovine gamma globulin 200 μl of 0.05 M Tris-HCl (pH 8.5) was added. At 4 ° C After incubating for 1 hour, the tube is centrifuged to determine the radioactivity in the pellet by gamma. It measured using the counter.Measurement of FSH bioactivity   Hormones: sheep FSH dissolved in PBS (NIH-FSH-SI Using R041212)), the physiological activity of FSH was measured.   Animals and treatments: Snell dwarf mice were isolated from conventional Holder et al. Described by J. Endocrinol., 85: 35-47 (1980). Propagated, maintained and marked as they were. 30 26-35 days old Female dwarf mice were arbitrarily divided into 5 treatment groups, with 0, 3.3, 10, and 30 respectively. , 90 μg / day oFSH subcutaneously twice daily for 5 days with 0.05 ml of solution Administered by injection. Fourteen hours after the last injection, mice were halothane / oxygen Killed by decapitation under anesthesia.   Ovarian and uterine weight: remove ovary and uterus and remove fat under stereomicroscope Dissected, fixed in formalin (3.7%), fixed, and cahn microbalance (Cahn microbalance).   Vaginal smear: a few drops of PB pulled out after being introduced into the vagina via a regular pipette S was used to collect vaginal cells. Transfer the cells onto a microscope slide and add methanol And stained with Wright-Giemsa. Analyze the smear under 100 times magnification, The conversion index [(keratinized cells / total cells) × 100] was calculated.   Statistical analysis: uterine and ovarian weights (expressed as mg / g body weight) and keratin Optimization index was used for statistical analysis. The data for each group treated with hormones Controls injected with PBS using Student's t-Test Compared with the rules.Promotion of FSH bioactivity using antiserum against sheep A1 peptide A   Hormone and purified antiserum: sheep FSH (NIH-FSH-SI) in PBS Was dissolved. Pre-immune serum or antiserum against peptide A Purified as follows: 1 volume of 27% sodium sulfate mixed with 2 volumes of serum , 37 ° C for 3.5 hours. Centrifuge the precipitated IG fraction and pellet Was reconstituted to the volume of serum to be induced. Next, this IG fraction was Highly dialyzed. Equivalent volumes of FSH and purified serum were injected to a final concentration of 100 μl 20 μg oFSH per solution and 50 μl of appropriately purified antiserum. Yeah. When using FSH alone, use PBS instead of serum However, when using only anti-peptide antiserum, use PBS instead of FSH. Was. Control animals received PBS only. 0.05m daily for mice 1 of the above solution (as indicated) was administered in two injections.   Animals and treatments: Dwarf mice are bred, maintained and marked as described above. I attached. Thirty 30 to 40 day old female Snell dwarf mice were arbitrarily treated with the following 5 treatments. Divided into groups: Group 1 (FSH): 20 g / mouse / day of oFSH dissolved in PBS; Group 2 (oFSH + antiserum; FSH + A): oFSH 20 μg + prepared as above Purified peptide A; 3 groups (oFSH + preimmune serum; FSH + P): 20 μg of oFSH + as above Prepared purified preimmune serum; Group 4 (antiserum alone; A): purified anti-peptide A antiserum prepared in the same manner as above + etc. Volume PBS; Group 5 (control; PBS): only PBS was administered.   Each animal was injected and sacrificed using the same protocol as in the previous experiment.   Uterine and ovarian weights (expressed as mg / g body weight) and keratinization index It measured similarly to the above. Differences between groups were determined by Student's t-test (Student 's t-Test).Promotion of FSH bioactivity using antiserum against peptide B   Animals and treatment: Twenty-five 31-40 day old female Snell dwarf mice were The lower 5 treatment groups were divided. Mice were bred and maintained as described above.   Hormones and purified antisera: hormones and sera were described in the previous section. It was prepared and purified in the same manner as described above. Group 1 (FSH): 20 g / mouse / day of oFSH dissolved in PBS; Group 2 (oFSH + antiserum; FSH + B): 20 μg oFSH + purified peptide B; 3 groups (oFSH + pre-immune serum; FSH + P): oFSH 20 μg + purified pre-immune blood Qing; Group 4 (antiserum alone; B): purified anti-peptide B antiserum alone; Group 5 (control; PBS): only PBS was administered.   Each animal was injected and sacrificed using the same protocol as in the previous experiment.   Uterine and ovarian weights (expressed as mg / g body weight) and vaginal smear The index of lamination was measured as described above. The difference between each group is Evaluation was made by Student's t-Test.result Production of antisera against peptides A, B, C, D and E   Figures 2a, b and c show in preimmune serum (bleed 0) and in pep. Immunized against two sheep (A1 and A2) injected with Pide A and against peptide B The antiserum (bleed (bleed)) of one of the two sheep treated sheep (B1) after four injections. 4) Comparison between antibody titers to bFSH in 4). Second Animal (B2) died before the end of the trial. Two sheep injected with peptide A Among the two, sheep A1 was the highest titer of antiserum that recognized bFSH. the above For antiserum from sheep (A1) and sheep B1 (injected with peptide B), In the nimbo We investigated the ability of oFSH to promote physiological effects in hypothermic snellen dwarf mice . In animals immunized with peptide C, D or E, almost all antisera titers were Not observed.oFSH bioactivity dose response curve   FIG. 3 shows the uterine and ovarian weights (expressed as mg / g body weight; FIGS. 3a and 3b). Of keratinization in relation to and in vaginal smears (% keratinized cells) 3c) shows the effect of varying amounts of oFSH alone. oFS Treatment with H significantly increased uterine weight in a dose-dependent manner (10 μg / day = 0.88 ± 0.19; P <0.05; 30 μg / day = 2.04 ± 0.34; P <0.01; 90 μg / day = 3.11 ± 0.18; P <0.00 1; significant values are for the PBS control group 0.34 ± 0.05). The ovaries weighed the two highest doses of oFSH (30 μg / day = 0.60 ± 0. 09; P <0.05; 90 μg / day = 1.18 ± 0.14; P <0.01). Control group (0.36 ± 0.05) was significantly larger than the weight However, a similar dose-dependent increase in ovarian weight was observed. Was. The keratinization index showed a similar pattern depending on the dose (1 0 μg / day = 13.48 ± 2.95; P <0.05; 30 μg / day = 31.85 ± 6.12; P <0.05; 90 μg / day = 59.89 ± 5.23; P <0.0 01; significant values are for the PBS control group 2.40 ± 1.56) .Promotion of FSH bioactivity by antiserum against peptide A   Figure 4 shows uterine (Fig. 4a) and ovarian (Fig. 4b) in Snell dwarf mice. Increased weight as well as increased keratinization index of vaginal smear (FIG. 4c). 5 shows the ability of the antiserum against peptide A to promote FSH activity.   With respect to uterine weight (FIG. 4a), antiserum against FSH + peptide A (FS H + A) response to FSH alone (FSH; P <0.001) or FSH + Significantly greater than observed with pre-immune serum (FSH + P; P <0.01) won. Significant between response to FSH alone and to FSH + pre-immune serum There was no significant difference. However, both responses show that antiserum alone against peptide A alone (A) or significantly greater than observed in controls injected with PBS Great (P <0.05 in all cases).   Similar results were obtained for ovarian weight (FIG. 4B). FSH + peptide The antiserum against A (FSH + A) is FSH alone (FSH; P <0.001). Or FSH + preimmune serum (FSH + P; P <0.01) It promoted a significantly greater increase in ovarian weight. FSH alone and FSH + immunity There was no significant difference between the responses to preserum; however, both responses were Antiserum alone against peptide A (A) or in controls injected with PBS Significantly greater than observed (P <0.05 in all cases).   Again, similar results were obtained for the vaginal smear keratinization index of the animals. (FIG. 4c). Against antiserum against FSH + peptide A (FSH + A) Responses were FSH alone (FSH; P <0.001) or FSH + pre-immune serum (FSH). SH + P; P <0.01). FSH only There was no significant difference between the responses to German and FSH + pre-immune sera. But However, both responses were due to antiserum alone against peptide A (A) or PBS injected. Significantly greater than that observed in the treated controls (in all cases) P <0.05).   All of these data are summarized in FIG. 4 and the associated tables.Promotion of FSH bioactivity by antiserum against peptide B   FIG. 5 shows the uterus (FIG. 5a) and ovary (FIG. 5b) in Snell dwarf mice. Increased weight as well as increased keratinization index of vaginal smear (FIG. 5c). 5 shows the ability of the antiserum against peptide B to promote FSH activity.   With respect to uterine weight (FIG. 5a), antiserum to FSH + peptide B (FS H + B), FSH alone (FSH; P <0.01) or FSH + Significantly greater than observed in preimmune serum (FSH + P; P <0.01) Was. Significant between responses to FSH alone and to FSH + pre-immune serum There was no difference. However, both responses show that antiserum alone against peptide B ( B) or significantly greater than observed in controls injected with PBS (P <0.01 in all cases).   Similar results were obtained for ovarian weight (FIG. 5B). FSH + peptide The antiserum against B (FSH + B) was FSH alone (FSH; P <0.01) or Eggs compared to those observed with FSH + pre-immune serum (FSH + P; P <0.01) It promoted a significantly greater increase in nest weight. FSH alone and FSH + pre-immune There were no significant differences between the responses to serum. FSH alone or FSH + pre-immune The response to serum was higher than that observed in controls injected with PBS. Significantly greater (P <0.05 in all cases).   Again, similar results were obtained for the vaginal smear keratinization index of the animals. (FIG. 5c). Against antiserum against FSH + peptide B (FSH + B) Responses were FSH alone (FSH; P <0.001) or FSH + pre-immune serum (FSH). SH + P; P <0.001). FSH Alone and against FSH + pre-immune serum There was no significant difference between the responses. Response to FSH alone, PBS injected Significantly greater than that observed in the treated control (P <0.05) .   All of these data are summarized in FIG. 5 and the associated tables.discussion   BFSH can be recognized by immunizing sheep with peptide A or B , But not with peptides C, D and E. To peptides No harmful physiologically affected sheep due to immune immunization Was. These results clearly show that either peptide A or peptide B Treatment of Hypopituitary Snell Dwarf Mice with FSH Mixed with Antisera Increased uterine and ovarian weight, measured by vaginal smear keratinization index It is shown to induce a dramatic enhancement of FSH bioactivity.   The use of peptide vaccines to produce FSH-enhancing antisera has other commercial importance. It has a number of advantages over methods of controlling the development of follicles of key species. Of these advantages Some of them are listed below. 1. The method uses a small amount of antigen to produce a relatively long-lasting effect.      Only temporary contact with the animal is required. 2. FSH-promoting antisera can be produced in test animals by active immunization;      Alternatively, an antibody preparation from one animal is injected into another animal of the same species by passive immunization.      May be given. 3. Using known FSH sequences from various species, specific peptide vaccines      Can be easily designed to suit a variety of commercially important animals. 4. The technology can be used to promote endogenous FSH activity;      Under the circumstances, the circulating level of FSH is under pituitary control      To continue, promotion occurs only when FSH is produced. As a result,      Ovarian hyperstimulation (hyperstimulati) often caused by existing superovulation procedures      on) could be prevented. 5. The active ingredients, both antiserum and FSH, are produced by animals.

[Procedure of Amendment] Article 184-8, Paragraph 1 of the Patent Act [Submission Date] September 15, 1997 (September 15, 1997) [Correction contents]                                The scope of the claims   1. A peptide having the following sequence capable of promoting FSH activity in a mammal:   (I) YTRDLVYRDPARPNI;   (Ii) YTRDLVYKDPARPNI;   (Iii) YTRDLVYKDPARPKI;   (Iv) RDPARPNIQKTC;   (V) KDPARPKIQKTC;   (Vi) KDPARPNIQKAC; or   (Vii) KDPARPNIQKTC, Or a sequence substantially homologous thereto.   2. A vaccine comprising one or more peptides according to claim 1.   3. Cattle, sheep, pigs, horses, humans, goats, camels, cats, dogs, llamas 3. The vaccine according to claim 2, wherein the vaccine is intended for administration to alpaca.   4. The vaccine according to claim 3, which is intended for administration to humans.   5. The mammal is a sheep, cow, horse, pig, human, camel, goat, cat, The peptide according to claim 1, which is a nu, a llama or an alpaca.   6. The peptide according to claim 5, wherein the mammal is a human.   7. 2. The method according to claim 1, which is used for promoting FSH activity in sheep. Peptide (i).   8. 2. The method according to claim 1, which is used for promoting FSH activity in sheep. Peptide (iv).   9. Claims 1 to be used for promoting bovine or porcine FSH activity. The peptide (i) described in the section. 10. Claims 1 to 5 for use in promoting FSH activity in horses or humans. The peptide (i) described in the section. 11. Claims used to promote FSH activity in sheep or pigs The peptide (ii) according to item 1. 12. Claims used to promote FSH activity in horses, humans or cattle Peptide (ii) according to box 1. 13. Claims 1 to 5 for use in promoting FSH activity in horses or humans. The peptide (iii) according to the above section. 14． Claims: Used to promote FSH activity in cattle, sheep or pig. The peptide (iii) according to item 1 of the scope. 15. 2. The method according to claim 1, which is used to promote bovine FSH activity. , Peptide (iv). 16. Claims used to promote FSH activity in pigs, horses or humans Peptide (iv) according to box 1. 17． Claims 1 to 5 for use in promoting FSH activity in horses or humans. Item (v). 18. Claims: Used to promote FSH activity in cattle, sheep or pig. The peptide (v) according to item 1 of the scope. 19. 2. The method according to claim 1, which is used for promoting FSH activity in sheep. Peptide (vi). 20. A contractor used to promote FSH activity in cattle, pigs, humans or horses. Peptide (vi) according to item 1 of the claim. 21. 2. The method according to claim 1, which is used for promoting FSH activity in pigs. , Peptide (vii). 22. Used to promote FSH activity in cattle, sheep, humans or horses; The peptide (vii) according to claim 1. 23. At least one peptide according to claim 1, and one or more pharmaceuticals A pharmaceutical formulation comprising a pharmaceutically acceptable carrier, excipient or diluent. 24. A peptide according to any one of claims 1 or 5 to 22. Or administering the pharmaceutical composition of claim 23 to a mammal. A method for promoting FSH activity in a mammal. 25. A peptide according to any one of claims 1 or 5 to 22. Or administering the pharmaceutical composition of claim 23 to a mammal. A method for improving fertility of a mammal. 26. The mammal is a sheep, cow, horse, pig, human, camel, goat, cat, 26. The method according to claim 24 or claim 25, which is a nu, a llama or an alpaca. Law. 27. 27. The method according to claim 26, wherein said mammal is a human. 28. Claim 1 or Claim 5 in the preparation of a medicament for promoting FSH activity. Use of the peptide according to any of paragraphs to 22.

────────────────────────────────────────────────── ─── Continuation of front page    (81) Designated countries EP (AT, BE, CH, DE, DK, ES, FI, FR, GB, GR, IE, IT, L U, MC, NL, PT, SE), OA (BF, BJ, CF) , CG, CI, CM, GA, GN, ML, MR, NE, SN, TD, TG), AP (KE, LS, MW, SD, S Z, UG), EA (AM, AZ, BY, KG, KZ, MD , RU, TJ, TM), AL, AM, AT, AU, AZ , BA, BB, BG, BR, BY, CA, CH, CN, CU, CZ, DE, DK, EE, ES, FI, GB, G E, HU, IL, IS, JP, KE, KG, KP, KR , KZ, LC, LK, LR, LS, LT, LU, LV, MD, MG, MK, MN, MW, MX, NO, NZ, P L, PT, RO, RU, SD, SE, SG, SI, SK , TJ, TM, TR, TT, UA, UG, US, UZ, VN (72) Inventor Beetle, James             United Kingdom, Isle Kay 65             Ciel, The Hanner Research Institute             Itit, (72) Inventor Holder, Andrew, Thomas             Cambridge Seabee 1 UK             2 BB, St. Bernabus Road               4

Claims (1)

[Claims]   1. A peptide having the following sequence:   (I) YTRDLVYRDPARPNI;   (Ii) YTRDLVYKDPARPNI;   (Iii) YTRDLVYKDPARPKI;   (Iv) RDPARPNIQKTC;   (V) KDPARPKIQKTC;   (Vi) KDPARPNIQKAC; or   (Vii) KDPARPNIQKTC, Or a sequence substantially homologous thereto.   2. A vaccine comprising one or more peptides according to claim 1.   3. Cattle, sheep, pigs, horses, humans, goats, camels, cats, dogs, llamas 3. The vaccine according to claim 2, wherein the vaccine is intended for administration to alpaca.   4. The vaccine according to claim 3, which is intended for administration to humans.   5. A nucleic acid sequence encoding the peptide according to claim 1 or a nucleic acid sequence encoding the same. A nucleic acid sequence having qualitative homology.   6. The nucleic acid sequence according to claim 5, which is a DNA sequence.   7. An antibody having specificity for one or more of the peptides of claim 1. .   8. The antibody according to claim 7, which is a monoclonal antibody.   9. An antibody preparation comprising the antibody according to claim 7 or 8. 10. 2. The method according to claim 1, which is used for promoting FSH activity in a mammal. Peptide. 11. The mammal is a sheep, cow, horse, pig, human, camel, goat, cat, The peptide according to claim 10, which is a nu, a llama or an alpaca. 12. 12. The peptide according to claim 11, wherein said mammal is a human. 13. 2. The method according to claim 1, which is used for promoting FSH activity in sheep. Peptide (i). 14． 2. The method according to claim 1, which is used for promoting FSH activity in sheep. Peptide (iv). 15. Claims 1 to be used for promoting bovine or porcine FSH activity. The peptide (i) described in the section. 16. Claims 1 to 5 for use in promoting FSH activity in horses or humans. The peptide (i) described in the section. 17． Claims used to promote FSH activity in sheep or pigs The peptide (ii) according to item 1. 18. Claims used to promote FSH activity in horses, humans or cattle Peptide (ii) according to box 1. 19. Claims 1 to 5 for use in promoting FSH activity in horses or humans. The peptide (iii) according to the above section. 20. Claims: Used to promote FSH activity in cattle, sheep or pig. The peptide (iii) according to item 1 of the scope. 21. 2. The method according to claim 1, which is used to promote bovine FSH activity. , Peptide (iv). 22. Claims used to promote FSH activity in pigs, horses or humans Peptide (iv) according to box 1. 23. Claims 1 to 5 for use in promoting FSH activity in horses or humans. Item (v). 24. Claims: Used to promote FSH activity in cattle, sheep or pig. The peptide (v) according to item 1 of the scope. 25. 2. The method according to claim 1, which is used for promoting FSH activity in sheep. Peptide (vi). 26. A contractor used to promote FSH activity in cattle, pigs, humans or horses. Peptide (vi) according to item 1 of the claim. 27. 2. The method according to claim 1, which is used for promoting FSH activity in pigs. , Peptide (vii). 28. Used to promote FSH activity in cattle, sheep, humans or horses; The peptide (vii) according to claim 1. 29. At least one peptide according to claim 1, and one or more pharmaceuticals A pharmaceutical formulation comprising a pharmaceutically acceptable carrier, excipient or diluent. 30. Peptide according to any one of claims 1 or 10 to 28. 10. The antibody according to claim 7 or 8, and the antibody according to claim 9 Administering to the mammal a body preparation, or a pharmaceutical formulation according to claim 29. A method for promoting FSH activity in a mammal. 31. Peptide according to any one of claims 1 or 10 to 28. 10. The antibody according to claim 7 or 8, and the antibody according to claim 9 Administering to the mammal a body preparation, or a pharmaceutical formulation according to claim 29. A method for improving fertility of a mammal, comprising: 32. The mammal is a sheep, cow, horse, pig, human, camel, goat, cat, 32. The method according to claim 30 or claim 31, which is a nu, a llama or an alpaca. Law. 33. 33. The method according to claim 32, wherein said mammal is a human. 34. Claim 1 in the preparation of a medicament that promotes FSH activity. Or a peptide according to any one of claims 10 to 28, wherein Use of the antibody according to claim 8, and the antibody preparation according to claim 9.