Claims for Patent: 5,798,266

Summary for Patent: 5,798,266

Title:	Methods and kits for obtaining and assaying mammary fluid samples for breast diseases, including cancer
Abstract:	Non-invasive methods and kits are provided for obtaining biological samples of mammary fluid or mammary fluid components by administering oxytocin or an oxytocin analogue to a mammalian patient to stimulate expression of mammary fluid. The oxytocin is preferably administered intranasally and causes myoepithelial contraction of target alveolar-ductal tissues of the breast. During or after mammary fluid expression, a biological sample is collected in the form of whole mammary fluid, whole cells or cellular components, other selected liquid or solid fractions of the mammary fluid, purified or bulk proteins, glycoproteins, peptides, nucleotides or other desired constituents of mammary fluid. Methods and kits are also provided for determining the presence or amount of a breast disease marker in biological samples of mammary fluid or mammary fluid components. Samples obtained according to the above sample collection methods are assayed to determine the presence and/or amount of a breast disease marker, for example a breast cancer marker such as CEA, HMFG, MCA, vasopressin, or cathepsin D, in the sample. Cellular samples obtained according to the above sample collection methods are also examined microscopically for cytological evidence of breast disease.
Inventor(s):	Quay; Steven C. (Edmonds, WA), Quay; Debra L. (Edmonds, WA)
Assignee:	K-Quay Enterprises, LLC (Edmonds, WA)
Application Number:	08/709,207
Patent Claims:	1. A non-invasive method for obtaining a biological sample from a mammary organ of a patient, comprising the steps of: administering to the patient an effective amount of a natural or synthetic oxytocin to stimulate expression of mammary fluid from a nipple of the patient; applying a breast pump to the nipple; and collecting a biological sample by exposing the expressed mammary fluid to a solid phase medium removably placed within said breast pump to retain a breast disease marker from the expressed mammary fluid. 2. The method for obtaining the biological sample according to claim 1, wherein the natural or synthetic oxytocin is administered by one or more modes of administration selected from the group consisting of intranasal administration, intravascular administration and intramuscular administration. 3. The method for obtaining the biological sample according to claim 1, wherein the natural or synthetic oxytocin is administered by intranasal administration. 4. The method for obtaining the biological sample according to claim 1, wherein the solid phase medium is selected from the group consisting of microscopic slides, membranes, filters and beads. 5. The method for obtaining the biological sample according to claim 1, wherein the step of administering involves administering an intranasal spray containing 40 USP units of oxytocin per ml in a liquid carrier. 6. The method for obtaining the biological sample according to claim 1, wherein the biological sample is selected from the group consisting of whole mammary fluid, whole cells, cell fragments, cell membranes, proteins, glycoproteins, peptides and nucleotide components of a primary mammary fluid sample. 7. The method for obtaining the biological sample according to claim 1, wherein the solid phase medium is a nitrocellulose filter. 8. An assay method for determining the presence or amount of a breast disease marker in a biological sample obtained from a mammary organ of a patient, comprising the steps of: administering to the patient an effective amount of a natural or synthetic oxytocin to stimulate expression of mammary fluid from a nipple of the patient; applying a sample collector to the nipple; and applying a breast pump to the nipple; and collecting a biological sample by exposing the expressed mammary fluid to a solid phase medium removably placed within said breast pump to retain a breast disease marker from the expressed mammary fluid; detecting the presence or amount of a breast disease marker in the biological sample. 9. The assay method according to claim 8, wherein the breast disease marker is a breast cancer marker. 10. The assay method according to claim 8, wherein the natural or synthetic oxytocin is administered by one or more modes of administration selected from the group consisting of intranasal administration, intravascular administration and intramuscular administration. 11. The assay method according to claim 8, wherein the natural or synthetic oxytocin is administered by intranasal administration. 12. The assay method according to claim 8, wherein the solid phase medium is selected from the group consisting of microscopic slides, membranes, filters and beads. 13. The assay method according to claim 8, wherein the step of administering oxytocin involves administering an intranasal spray containing 40 USP units of oxytocin per ml in a liquid carrier. 14. The assay method according to claim 8, wherein the biological sample is selected from the group consisting of whole mammary fluid, whole cells, cell fragments, cell membranes, purified proteins, bulk proteins, glycoproteins, peptides and nucleotide components of a primary mammary fluid sample. 15. The assay method according to claim 8, wherein the breast disease marker is selected from the group consisting of a protein, a peptide, a glycoprotein, a lipid, a DNA polynucleotide and an RNA polynucleotide. 16. The assay method according to claim 8, wherein the breast disease marker is detected by employing an immunological probe that specifically binds the marker. 17. The assay method according to claim 16, wherein the immunological probe is a monoclonal antibody. 18. The assay method according to claim 8, wherein the breast disease marker comprises a cytological phenotype of cells, cell clusters or cell fragments. 19. The assay method according to claim 8, wherein a plurality of different breast disease markers are detected using a panel of immunological probes. 20. The assay method according to claim 8, wherein the breast disease marker is a breast cancer marker selected from the group consisting of a DNA polynucleotide and an RNA polynucleotide, and wherein the step of detecting the presence or amount of the breast cancer marker in the biological sample employs a nucleotide probe. 21. The assay method according to claim 8, wherein the breast disease marker is selected from the group consisting of Ki67 Growth Factor, Cyclin D1, Proliferating Cell Nuclear Antigen, Transforming Growth Factor .alpha., Tissue Plasminogen Activator, Insulin Growth Factor Receptors, Collagenase Type IV, Laminin Receptor, Integrins, p53, rb, nm23, ras, c-myc, Heat Shock Proteins, Prolactin, Neuron-Specific Enolase, IR-14, KA 1, KA 14, Alpha-Lactalbumin and Actin. 22. The assay method according to claim 8, wherein the breast disease marker is selected from the group consisting of CEA, HMFG, MCA, vasopressin and cathepsin D. 23. The assay method according to claim 8, wherein the solid phase medium is a nitrocellulose filter. 24. A kit for determining the presence or amount of a breast disease marker in a biological sample obtained from a mammary organ of a patient, comprising: a pharmaceutical preparation of a natural or synthetic oxytocin in a biologically suitable carrier; collecting means for collecting a biological sample from expressed mammary fluid, said collecting means comprising a solid phase device selected from the group consisting of microscopic slides, membranes, filters and beads; and detecting means for detecting the presence and/or amount of a breast disease marker in said biological sample. 25. The kit according to claim 24, wherein the oxytocin preparation is provided in an intranasal spray applicator and contains approximately 40 USP units of oxytocin per ml of a liquid carrier. 26. The kit according to claim 24, wherein the collecting means is fluidly connected to a breast pump applicable to a human breast and designed to generate sustained negative pressures in an area surrounding a nipple of between about 50-200 mm Hg. 27. The kit according to claim 26, wherein the collecting means is incorporated within the breast pump. 28. The kit according to claim 24, wherein the detecting means includes an immunological probe that specifically binds the breast disease marker. 29. The kit according to claim 28, wherein the immunological probe is a monoclonal antibody. 30. The kit according to claim 24, wherein the detecting means includes a panel of immunological probes that collectively bind to a plurality of different breast disease markers. 31. The kit according to claim 24, wherein the detecting means includes a nucleotide probe that specifically hybridizes to a breast disease marker selected from the group consisting of a DNA polynucleotide and an RNA polynucleotide. 32. The kit according to claim 24, wherein the detecting means includes an immunological probe that specifically binds to one or more breast cancer markers selected from the group consisting of Ki67 Growth Factor, Cyclin D1, Proliferating Cell Nuclear Antigen, Transforming Growth Factor .alpha., Tissue Plasminogen Activator, Insulin Growth Factor Receptors, Collagenase Type IV, Laminin Receptor, Integrins, p53, rb, nm23, ras, c-myc, Heat Shock Proteins, Prolactin, Neuron-Specific Enolase, IR-14, KA 1, KA 14, Alpha-Lactalbumin and Actin. 33. The kit according to claim 24, wherein the detecting means includes an immunological probe that specifically binds to one or more breast cancer markers selected from the group consisting of CEA, HMFG, MCA, vasopressin and cathepsin D. 34. A kit for collecting a biological sample from a mammary organ of a patient, comprising: a breast pump; and a solid phase sample collection medium selected from the group consisting of membranes, filters and beads for placement in fluid connection with said breast pump to contact the biological sample and retain a breast cancer marker. 35. The kit according to claim 34, wherein the solid phase sample collection medium is a nitrocellulose filter. 36. The kit of claim 34, wherein the solid phase sample collection medium is incorporated within the breast pump. 37. The kit of claim 34, wherein the solid phase sample collection medium is removably placed within the breast pump.

Details for Patent 5,798,266

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Smith & Nephew, Inc.	SANTYL	collagenase	Ointment	101995	06/04/1965		2039-02-26