Disclosed are 2-methylene analogs of vitamin D.sub.3 and related compounds, their biological activities, and various pharmaceutical uses for these analogs. Particularly disclosed are 1-hydroxy-2-methylene-vitamin D.sub.3, (20S)-1-hydroxy-2-methylene-vitamin D.sub.3, and (5E)-1,25-dihydroxy-2-methylene-vitamin D.sub.3, their biological activities, and various pharmaceutical uses for these compounds including methods of treating and/or preventing bone diseases and disorders.

Disclosed are 2-methylene analogs of vitamin D.sub.3 and related compounds, their biological activities, and various pharmaceutical uses for these analogs. Particularly disclosed are 1-hydroxy-2-methylene-vitamin D.sub.3, (20S)-1-hydroxy-2-methylene-vitamin D.sub.3, and (5E)-1,25-dihydroxy-2-methylene-vitamin D.sub.3, their biological activities, and various pharmaceutical uses for these compounds including methods of treating and/or preventing bone diseases and disorders.

Methods include determining an amount of vitamin D analyte in a sample suspected of containing the vitamin D analyte. A combination is provided in an assay medium that includes the sample, a vitamin D epimer antibody that is specific for epimers of the vitamin D analyte wherein the vitamin D epimer antibody does not bind to any detectable degree to the vitamin D analyte, and a vitamin D antibody that is specific for the vitamin D analyte, and. The assay medium is incubated under conditions for binding of the vitamin D epimer antibody to the epimers of the vitamin D analyte and for binding of the vitamin D antibody to the vitamin D analyte to form a vitamin D antibody-bound complex. The amount of vitamin D antibody-bound complex is determined and related to the amount of vitamin D analyte in the sample.

The invention provides non-hormonal vitamin D conjugated to apelin proteins that result in increased absorption, bioavailability or circulating half-life when compared to non-conjugated forms. In some embodiments, the vitamin D targeting groups are coupled to the apelin proteins via the third carbon on the vitamin D backbone.

According to the present invention, there can be provided a process for producing 7-dehydrocholesterol (7DHC), comprising culturing, in a medium, a 7DHC-producing Labyrinthulea microorganism in which sterol 24-C-methyltransferase activity is reduced or lost as compared to a parent strain, allowing 7DHC to be produced and accumulated in the culture, and collecting the 7DHC from the culture; and a process for producing vitamin D3, comprising irradiating, with ultraviolet light, 7-dehydrocholesterol produced by the production process.

According to the present invention, there can be provided a process for producing 7-dehydrocholesterol (7DHC), comprising culturing, in a medium, a 7DHC-producing Labyrinthulea microorganism in which sterol 24-C-methyltransferase activity is reduced or lost as compared to a parent strain, allowing 7DHC to be produced and accumulated in the culture, and collecting the 7DHC from the culture; and a process for producing vitamin D3, comprising irradiating, with ultraviolet light, 7-dehydrocholesterol produced by the production process.

Disclosed are 2-methylene analogs of vitamin D.sub.3 and related compounds, their biological activities, and various pharmaceutical uses for these analogs. Particularly disclosed are 1-hydroxy-2-methylene-vitamin D.sub.3, (20S)-1-hydroxy-2-methylene-vitamin D.sub.3, and (5E)-1,25-dihydroxy-2-methylene-vitamin D.sub.3, their biological activities, and various pharmaceutical uses for these compounds including methods of treating and/or preventing bone diseases and disorders.

Methods include determining an amount of vitamin D analyte in a sample suspected of containing the vitamin D analyte. A combination is provided in an assay medium that includes the sample, a vitamin D epimer antibody that is specific for epimers of the vitamin D analyte wherein the vitamin D epimer antibody does not bind to any detectable degree to the vitamin D analyte, and a vitamin D antibody that is specific for the vitamin D analyte, and. The assay medium is incubated under conditions for binding of the vitamin D epimer antibody to the epimers of the vitamin D analyte and for binding of the vitamin D antibody to the vitamin D analyte to form a vitamin D antibody-bound complex. The amount of vitamin D antibody-bound complex is determined and related to the amount of vitamin D analyte in the sample.

Methods include determining an amount of an epimeric vitamin D analyte in a sample suspected of containing the epimeric vitamin D analyte. A combination is provided in an assay medium that includes the sample and a vitamin D epimer antibody that is specific for the epimeric vitamin D analyte. The assay medium is incubated under conditions for binding of the vitamin D epimer antibody to the epimeric vitamin D analyte to form an epimeric vitamin D antibody-bound complex. The amount of the epimeric vitamin D antibody-bound complex is determined and related to the amount of epimeric vitamin D analyte in the sample.

The present invention relates to new vitamin D compounds which are bonded to a labelling group at the C3 stereocentre by means of a linker. The present invention further relates to a method for producing these vitamin D compounds and to the use of an intermediate for producing these compounds. The present invention also relates to a method for quantitatively determining vitamin D using a vitamin D compound according to the invention as a tracer. Furthermore, the present invention relates to a reagent for determining vitamin D, containing a compound according to the invention, and to the use thereof for determining vitamin D.