Find Funding

Contract Topics

230 Synthesis of Stable Isotope-Labeled Steroids as Internal Standards for the Measurement of Endogenous Steroid Hormones in Biologic Samples by Liquid Chromatography - Mass Spectrometry (LC-MS)

Number of anticipated awards: 1-3
(Fast-Track proposals in response to this solicitation are encouraged.)
Budget (total costs): Phase I: $150,000;
Phase II: $750,000

The deadline for receipt of all contract proposals submitted in response to this solicitation was:
5:00 p.m. Eastern Standard Time
Monday, November 6, 2006

Short-term project goals: To develop chemical syntheses for specific steroids labeled with stable isotopes and to demonstrate that the products meet defined criteria for use as internal standards in LC-MS measurement of endogenous steroid hormones.

Long-term project goals: To scale up the chemical syntheses of these stable isotope-labeled steroids and combine them in one or more kits. These stable isotope internal standards will permit academic and clinical laboratories to establish sensitive, accurate LC-MS methods for measuring endogenous steroid hormones in biologic samples and to standardize the measurements across laboratories.

Although endogenous steroid hormones, specifically estrogens, androgens, and progestogens, are believed to play critical roles in the etiology of breast, endometrial, ovarian, prostate, and possibly other cancers, research has been limited for many years by the absence of sensitive, accurate methods for measuring the absolute concentrations of steroid hormones and their metabolites in serum, urine, and tissue. The assays routinely used by epidemiologists and clinicians have relied on radiobinding kits with poor specificity and sensitivity in biologic matrixes. Many hypotheses about specific hormone metabolites, genetically-determined patterns of hormone metabolism, and their contributions to cancer prevention, causation, and treatment could not be evaluated. In a multidisciplinary collaboration between the NCI Division of Cancer Epidemiology and Genetics, the NCI Center for Cancer Research, and SAIC-Frederick, LC-MS methods for measuring endogenous estrogens, androgens, and progestogens in urine, blood, and tissue are being developed. The methods are sensitive, accurate, precise, robust, and sufficiently rapid and inexpensive to permit the measurement of endogenous hormones in the multiple biologic specimens collected in epidemiologic and clinical research.

The collaboration has led to the development of a patent-pending, validated high performance liquid chromatography-electrospray ionization-tandem mass spectrometry method for measuring the absolute quantities of 15 endogenous estrogens and their metabolites in human urine. It requires only 0.5 mL of urine, yet is capable of simultaneously quantifying all urinary estrogens and their metabolites in premenopausal and postmenopausal women and men. Details of the current analytical method, including the identities of the five isotopically labeled standards already being utilized, can be found in Xu et al, Anal Chem 2005;77:6646-54. The method is now being applied to serum and plasma, anticipating that it will be able to measure all free and conjugated estrogens.

The National Cancer Institute is seeking a contract(s) with a small business(es) capable of preparing pure samples of steroid hormones and steroid hormone metabolites that are labeled with deuterium, C-13, or O-17. It is expected that these labeled steroids will be commercialized by the small business for use as internal standards in LC-MS measurement techniques. The steroid internal standards will be used to ensure accurate, precise, sensitive measurements from LC-MS machines in different laboratories and to minimize intra- and inter-laboratory variability of results.

With these methods, the scientific community will be able to ask important questions about the roles of endogenous estrogens, androgens, and progestogens in the initiation and progression of specific cancers. This method will also be relevant to cancer prevention and treatment. For example, women at high risk of breast cancer could be identified because of inherited or environmentally-induced estrogen metabolism patterns. In addition, the monitoring the effects of various estrogen binding antagonists and estrogen synthesis inhibitors currently being used in breast cancer treatment could be achieved.

Phase I Activities and expected deliverables:

In Phase I, the small business(es) will demonstrate the feasibility of their process. The small business will demonstrate that they can synthesize at least five stable isotope-labeled steroids of those listed below, provide full mass spectra that demonstrate that each of the steroids meet the criteria below, and provide samples of each steroid so that NCI can confirm structure, purity, and stability. The following compounds are of interest:

• estrone, 2-hydroxyestrone, 4-hydroxyestrone, 16a-hydroxyestrone, 2-methoxyestrone, 3-methoxyestrone, 4-methoxyestrone, 17-ß-estradiol, 2-hydroxyestradiol, 4-hydroxyestradiol, 2-methoxyestradiol, 4-methoxyestradiol, 16-keto-17ß-estradiol, estriol, 16-epiestriol, 17-epiestriol, 2-hydroxyestrone-3-methyl ether, estrone-3-sulfate, androstenedione, testosterone, dehydroepiandrosterone sulfate, dehydroepiandrosterone, and progesterone.
• Each must be labeled with deuterium, C-13, or O-17; the label can not be placed in the sulfate moiety.
• A full mass spectrum should accompany each preparation delivered and should confirm that no more than 2% of the total mass in the molecular ion region appears at molecular weights smaller than that of the natural material plus 3.
• In addition, a full mass spectrum should demonstrate that even after dissolution in a pH 9 aqueous medium for 10 minutes at 60o C., no more than 2% of the total mass in the molecular ion region appears at molecular weights smaller than that of the natural material plus 3.
• Additional information regarding these specifications is available by contacting:
  
  
  Mary E. Landi O'Leary
  Contracting Officer
  Procurement Analyst
  National Cancer Institute
  EPS 6120, Suite 600, Room 6044
  MSC-7192
  Phone: (301) 435-3807
  FAX: (301) 480-0309
  Phone: (301) 435-3783
  Email: ml186r@nih.gov
  

Phase II Activities and expected deliverables:

• The synthetic methods developed for Phase I must be scaled up for producing at least 0.1 g of each stable isotope-labeled steroid. The larger quantities must meet the criteria listed in Phase I above. Full mass spectra will be required as documentation.
• One or more kits containing stable isotope-labeled steroids will be designed and produced. The kits should allow for an interchangeable suite of isotope-labeled steroids, per the needs of the scientific community.
• Instructions for use of the standards kit.