Drug – bio-affecting and body treating compositions – Designated organic active ingredient containing – Peptide containing doai
Patent
1996-01-05
1998-12-15
Russel, Jeffrey E.
Drug, bio-affecting and body treating compositions
Designated organic active ingredient containing
Peptide containing doai
435806, 436 63, 436906, 530300, 530344, 530350, 530417, 530419, 530850, A61K 3802, C07K 120, C07K 1447, G01N 3348
Patent
active
058497137
DESCRIPTION:
BRIEF SUMMARY
FIELD AND BACKGROUND OF THE INVENTION
The present invention relates generally to the field of human fertilization, and more particularly to human factors purifiable from human follicular fluid exerting both chemotactic and chemokinetic activities on human spermatozoa. The invention further concerns various treatment and diagnostic procedures related to human fertilization.
The problem of infertility afflicts about 10% of all couples. Of these, about 40-50% are related to female disorders and about 40% to male disorders, the remaining 10-20% of the cases lacking explanation. There is an ever growing number of procedures for the diagnosis and treatment of female sterility, while means for the treatment of male infertility or of infertility caused by defects in the sperm-egg interactions are quite limited.
One of the questions in mammalian fertilization, in general, and in human fertilization, in particular, was whether the egg communicates with spermatozoa in the female genital tract before the actual contact between them, or whether the sperm-egg contact is coincidental. A common mode of communication in nature is chemotaxis (i.e., the response of motile cells or organisms to the gradient of a chemical stimulus, resulting in modulation of the direction of travel so as to approach an attractant or to move away from a repellent), often accompanied by chemokinesis (i.e., a change in swimming speed in response to a chemical stimulus). Sperm chemotaxis is known to occur in species with external fertilization, primarily in metazoa whose females spawn their eggs into sea water before fertilization. Animal sperm attractants are peptides or proteins with molecular sizes varying from 1.5 kDa for sea urchin sperm attractant to 25 kDa for the sperm attractant of the siphonophore Muggiaea atlantica. Sperm chemotaxis is less obvious in species with internal fertilization, where very large numbers of spermatozoa (ca. 10.sup.7 -10.sup.9 in mammals) are ejaculated directly into the female reproductive tract; there, a sufficient number may reach the egg coincidentally, and a need for a directed movement or for sperm chemotaxis is not self-evident. However, recent studies indicate that sperm chemotaxis is also present in mammals.
Follicular fluids have a large number of substances, such as steroids, lipids, lipid proteins, peptide hormones, growth factors, etc., that affect sperm functions such as motility and metabolism.
In a previous study (Ralt et al., 1991), the inventors have shown that human spermatozoa accumulated in follicular fluid in vitro and that sperm attraction to follicular fluid correlated with egg fertilizability. It was thus suggested that the follicular fluid may contain a factor(s) released by the egg or its surrounding cells that is capable of attracting sperm. However, no factor has been isolated and characterized in this previous study, nor was the nature of the attraction identified.
SUMMARY OF THE INVENTION
It has now been found, in accordance with the present invention, that by precipitation of human follicular fluid with a protein-precipitating organic solvent, e.g. 90% acetone, the sperm chemotactic activity is found in the supernatant and not in the precipitate. Further fractionation of the acetone supernatant by size-exclusion chromatography followed by reversed-phase chromatography lead to two active fractions of peptidic nature exhibiting sperm chemotaxis--one containing a chemotactic factor with molecular size of about 13 kDa and the other a chemotactic factor with apparent molecular size smaller than 1.3 kDa (<1.3 kDa factor, herein).
The molecular size of the chemotactic factors was determined by high pressure gel filtration using a Bio-Sil TSK-125 600.times.7.5 mm column (Bio Rad, CA) and elution with 100 mM phosphate-buffered saline (PBS) (pH 7.4) at room temperature. Two fractions exhibiting sperm chemotactic activity were obtained: a first active fraction eluted at 17.0-17.5 min corresponding to molecular size of 13.+-.1 kDa and a second fraction eluted at 25.0-25.5 min after the
REFERENCES:
patent: 4767703 (1988-08-01), Ax et al.
patent: 4772554 (1988-09-01), Ax et al.
Manor et al, Purification of Chemotactic Factors . . . Molecular Biology of the Cell, Oct. 1993, vol. 4, p. 139a, Abstr. 809.
Ralt et al. Sperm Attraction to a Follicular Factors(s) Proc. Natl. Acad. Sci. Apr. 1991, vol. 88, pp. 2840-2844.
Lee, Shuang-Lin, Yu-Min Kuo, Chin-Cheng Kao, Chuang-Ye Hong, and Yau-Huei Wei; "Purification of a Sperm Motility Stimulator from Porcine Follicular Fluid"; Comparitive Biochemistry and Physiology; vol. 101B, No. 4; pp. 591-594; 1992.
Lachapelle. MH, R. Bouzayen, J. Langlais, K. Jarvi, J. Bourque, and P. Miron; "Effect of Lysoplatele-Activating Factor on Human Sperm Fertilizing Ability"; Fertil Steril; vol. 59, No. 4; Abstract; Apr. 1993.
Giorgetti C., E. Hans, J.L. Spach, P. Auquier, and R. Roulier; "In-Vitro Fertilization in Cases with Severe Sperm Defect Use of a Swim-Across Technique and Medium Supplemented with Follicular Fluid"; Human Reproduction; vol. 7, No. 8; Abstract; 1992.
Russel Jeffrey E.
Yeda Research and Development Co. Ltd.
LandOfFree
Chemotactic factors for human spermatozoa and their use in human does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Chemotactic factors for human spermatozoa and their use in human, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Chemotactic factors for human spermatozoa and their use in human will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-1457966