# Patent

## Title
Crystallization a 1alpha-hydroxy-20-methyl-2-methylene-19,24,25,26,27-pentanorvitamin D3

## Abstract

A method of purifying 1α-hydroxy-20-methyl-2-methylene-19,24,25,26,27-pentanorvitamin D3 to obtain 1α-hydroxy-20-methyl-2-methylene-19,24,25,26,27-pentanorvitamin D3 in crystalline form. The method includes the steps of preparing a solvent of either ethyl formate or a mixture of ethyl formate and hexane, dissolving a product containing 1α-hydroxy-20-methyl-2-methylene-19,24,25,26,27-pentanorvitamin D3 to be purified in the solvent, cooling the solvent and dissolved product below ambient temperature for a sufficient amount of time to form a precipitate of 1α-hydroxy-20-methyl-2-methylene-19,24,25,26,27-pentanorvitamin D3 crystals, and recovering the 1α-hydroxy-20-methyl-2-methylene-19,24,25,26,27-pentanorvitamin D3 crystals.

## Background
CROSS REFERENCE TO RELATED APPLICATIONS
This application claims the benefit of U.S. Provisional Application No. 61/529,007, filed Aug. 30, 2011, which is incorporated by reference herein in its entirety for any purpose.


STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT
This invention was made with government support under DK047814 awarded by the National Institutes of Health. The government has certain rights in the invention.


BACKGROUND OF THE INVENTION
The present invention relates to purification of organic compounds, and more particularly to the purification of 1α-hydroxy-20-methyl-2-methylene-19,24,25,26,27-pentanorvitamin D3 (referred to herein as “20DCM”) by preparing it in crystalline form.
Purification of organic compounds, especially those designated for pharmaceutical use, is of considerable importance for chemists synthesizing such compounds. Preparation of the compound usually requires many synthetic steps and, therefore, the final product can be contaminated not only with side-products derived from the last synthetic step of the procedure but also with compounds that were formed in previous steps. Even chromatographic purification, which is a very efficient but relatively time-consuming process, does not usually provide compounds which are sufficiently pure to be used as drugs.
Depending on the method used to synthesize 1α-hydroxyvitamin D compounds, different minor undesirable compounds can accompany the final product. Thus, for example, if direct C-1 hydroxylation of 5,6-trans geometric isomer of vitamin D is performed, followed by SeO2/NMO oxidation and photochemical irradiation [see Andrews et al., J. Org. Chem. 51, 1635 (1986); Calverley et al., Tetrahedron 43, 4609 (1987); Choudry et al, J. Org. Chem. 58, 1496 (1993)], the final 1α-hydroxyvitamin D product can be contaminated with 1β-hydroxy- as well as 5,6-trans isomers. If the method consists of C-1 allylic oxidation of the 4-phenyl-1,2,4-triazoline-3,5-dione adduct of the previtamin D compound, followed by cycloreversion of the modified adduct under basic conditions [Nevinckx et al., Tetrahedron 47, 9419 (1991); Vanmaele et al, Tetrahedron 41, 141 (1985) and 40, 1179 (1994); Vanmaele et al., Tetrahedron Lett. 23. 995 (1982)], one can expect that the desired 1α-hydroxyvitamin can be contaminated with the previtamin 5(10), 6,8-triene and 1β-hydroxy isomer. One of the most useful C-1 hydroxylation methods, of very broad scope and numerous applications, is the experimentally simple procedure elaborated by Paaren et al. [see J. Org. Chem. 45, 3253 (1980) and Proc. Natl. Acad. Sci U.S.A. 75, 2080 (1978)]. This method consists of allylic oxidation of 3,5-cyclovitamin D derivatives, readily obtained from the buffered solvolysis of vitamin D tosylates, with SeO2/t-BuOOH and subsequent acid-catalyzed cycloreversion to the desired 1α-hydroxy compounds. Taking into account this synthetic path it is reasonable to assume that the final product can be contaminated with 1α-hydroxy epimer, 5,6-trans isomer and the previtamin D form. 1α-hydroxyvitamin D4 is another undesirable contaminant found in 1α-hydroxyvitamin D compounds synthesized from vitamin D2 or from ergosterol. 1α-hydroxyvitamin D4 results from C-1 oxidation of vitamin D4, which in turn is derived from contamination of the commercial ergosterol material. Typically, the final product may contain up to about 1.5% by weight 1α-hydroxyvitamin D4. Thus, a purification technique that would eliminate or substantially reduce the amount of 1α-hydroxyvitamin D4 in the final product to less than about 0.1-0.2% would be highly desirable.
The vitamin D conjugated triene system is not only heat- and light-sensitive but it is also prone to oxidation, leading to the complex mixture of very polar compounds. Oxidation usually happens when a vitamin D compound has been stored for a prolonged time. Other types of processes that can lead to a partial decomposition of vitamin D compounds consist of some water-elimination reactions; their driving force is allylic (1α-) and homoallylic (3β-) position of the hydroxy groups. The presence of such above-mentioned oxidation and elimination products can be easily detected by thin-layer chromatography.
Usually, all 1α-hydroxylatation procedures require at least one chromatographic purification. However, even chromatographically purified 1α-hydroxyvitamin D compounds, although showing consistent spectroscopic data, suggesting homogeneity, do not meet the purity criteria required for therapeutic agents that can be orally, parenterally or transdermally administered. Therefore, it was evident that a suitable method of purification of the 1α-hydroxylated vitamin D compound 20DCM is required.
SUMMARY OF THE INVENTION
The present invention relates to a method of purifying 20DCM by means of crystallization to obtain 20DCM in crystalline form. The solvent plays a crucial role in the crystallization process, and is typically an individual liquid substance or a suitable mixture of different liquids. For crystallizing 20DCM, the most appropriate solvent and/or solvent system is characterized by the following factors:

    
    
        (1) low toxicity;
        (2) low boiling point;
        (3) significant dependence of solubility properties with regard to temperature (condition necessary for providing satisfactory crystallization yield); and
        (4) relatively low cost.
    
    


Interestingly, hexane, so frequently used for crystallization purposes, was found less suitable as the sole solvent for crystallization of 20DCM. However, it was found that either ethyl formate, or a mixture of ethyl formate and hexane, was most useful for the crystallization of 20DCM. In particular, it was determined that a mixture of about 75% ethyl formate with about 25% hexane (by volume) performed well. The ethyl formate/hexane solvent mixture was also easy to remove by evaporation or other well known methods. In all cases the crystallization process occurred easily and efficiently; and the precipitated crystals were sufficiently large to assure their recovery by filtration or other means.



BRIEF DESCRIPTION OF THE DRAWINGS
FIG. 1 is an illustration of the three dimensional molecular structure for 20DCM as defined by the atomic positional parameters discovered and set forth herein;



DETAILED DESCRIPTION OF THE INVENTION
The present invention provides 1α-hydroxy-20-methyl-2-methylene-19,24,25,26,27-pentanorvitamin D3 (20DCM) in crystalline form, a pharmacologically important compound, characterized by the formula I shown below:




The present invention also provides a valuable method of purification of 20DCM. The purification technique involves obtaining the 20DCM product in crystalline form by utilizing a crystallization procedure wherein the 20DCM material to be purified is dissolved using as the solvent either ethyl formate as the sole solvent, or a mixture comprised of ethyl formate and hexane. Preferably the mixture comprises about 75% ethyl formate and about 25% hexane (by volume). Thereafter, the solvent can be removed by evaporation, with or without vacuum, or other means as is well known, or the resultant crystals may be filtered from the mother liquor. The technique can be used to purify a wide range of final products containing 20DCM obtained from any known synthesis thereof, and in varying concentrations, i.e. from microgram amounts to kilogram amounts. As is well known to those skilled in this art, the amount of solvent utilized should be minimized and/or adjusted according to the amount of 20DCM to be purified.
The usefulness and advantages of the present crystallization procedure is shown in the following specific Examples 1, 2 and 3. After crystallization, the precipitated material was observed under a microscope to confirm its crystalline form. Yields of crystals were relatively high and the obtained crystals showed a relatively sharp melting point of 140-145° C.
The described crystallization process of the synthetic 20DCM product represents a valuable purification method, which can remove most side products derived from the synthetic path. Such impurity is the result of the contamination of starting raw materials. The crystallization process occurred easily and efficiently; and the precipitated crystals were sufficiently large to assure their recovery by filtration, or other means.
Crystallization of 1α-hydroxy-20-methyl-2-methylene-19,24,25,26,27-pentanorvitamin D3 (20DCM)
EXAMPLE 1
Crystallization from Ethyl Formate
1α-Hydroxy-20-methyl-2-methylene-19,24,25,26,27-pentanorvitamin D3, 20DCM (15 mg), was dissolved in boiling ethyl formate (0.35 mL) and left at room temperature for about 1 hour, then it was kept in a refrigerator for about 18 hours. The precipitated crystals were filtered off, washed with a small volume of a cold (0° C.) ethyl formate and dried to give 9 mg (60%) of crystalline material.
EXAMPLE 2
Crystallization from Ethyl Formate/Hexane
1α-Hydroxy-20-methyl-2-methylene-19,24,25,26,27-pentanorvitamin D3, 20DCM (15 mg), was dissolved in boiling ethyl formate (0.30 mL) and hexane (0.10 mL) was added. It was left at room temperature for about 1 hour, then it was kept in a refrigerator for about 18 hours. The precipitated crystals were filtered off, washed with a small volume of a cold (0° C.) ethyl formate/hexane (3:1) mixture and dried to give 10 mg (67%) of crystalline material.
EXAMPLE 3
Experimental
All crystal measurements were performed on a KM4CCD κ-axis diffractometer with graphite-monochromated MoKα radiation. The crystal was positioned at 62 mm from the CCD camera. 2186 frames were measured at 0.5° intervals with a counting time of 12 sec. The data were corrected for Lorentz and polarization effects. Empirical correction for absorption was applied[1] Data reduction and analysis were carried out with the Oxford Diffraction programs.[2]
The structure was solved by direct methods[3] and refined using SHELXL.[4] The refinement was based on F2 for all reflections except those with very negative F2. Weighted R factors wR and all goodness-of-fit S values are based on F2. Conventional R factors are based on F with F set to zero for negative F2. The Fo2>2σ(Fo2) criterion was used only for calculating R factors and is not relevant to the choice of reflections for the refinement. The R factors based on F2 are about twice as large as those based on F. All hydrogen atoms were located geometrically and their position and temperature factors were not refined. Scattering factors were taken from Tables 6.1.1.4 and 4.2.4.2 in Reference 5.
The three dimensional structure of 20DCM as defined by the following physical data and atomic positional parameters described and calculated herein is illustrated in FIG. 1.







TABLE 1





Crystal data and structure refinement for 20DCM.


















Identification code
20dcm



Empirical formula
C24H38O2



Formula weight
358.54



Temperature
100(2) K



Wavelength
0.71073 A



Crystal system, space group
Orthorhombic, P2(1)2(1)2(1)



Unit cell dimensions:



a = 6.6085(2) A
alpha = 90 deg.



b = 15.8069(5) A
beta = 90 deg.



c = 20.0641(7) A
gamma = 90 deg.



Z, Calculated density
4, 1.136 Mg/m{circumflex over ( )}3



Absorption coefficient
0.070 mm{circumflex over ( )}−1



F(000)
792



Crystal size
0.70 × 0.40 × 0.35 mm



Theta range for data collection
2.77 to 28.73 deg.



Limiting indices
−8 <= h <= 8, −20 <= k <= 21,




−26 <= 1 <= 26



Reflections collected/unique
35848/3009 [R(int) = 0.0196]



Completeness to theta
= 28.00 99.7%



Absorption correction
Semi-empirical from equivalents



Max. and min. transmission
0.98 and 0.96



Refinement method
Full-matrix least-squares on F{circumflex over ( )}2



Data/restraints/parameters
3009/13/261



Goodness-of-fit on F{circumflex over ( )}2
1.084



Final R indices [I > 2sigma(I)]
R1 = 0.0412, wR2 = 0.1103



R indices (all data)
R1 = 0.0479, wR2 = 0.1136



Largest diff. peak and hole
0.217 and −0.256 e · A{circumflex over ( )}−3

















TABLE 2







Atomic coordinates (×104) and equivalent isotropic displacement


parameters (A2 × 103) for 20DCM. U(eq) is defined as one third of the


trace of the orthogonalized Uij tensor.












x
y
z
U(eq)

















O(1)
8264(2)
−1494(1)
9984(1)
30(1)



O(2)
2152(2)
−1902(1)
10503(1)
35(1)



C(1)
6215(2)
−1295(1)
10144(1)
26(1)



C(2)
5551(3)
−1666(1)
10802(1)
33(1)



C(2′)
6749(3)
−2110(2)
11197(1)
54(1)



C(3)
3382(3)
−1462(1)
10976(1)
34(1)



C(4)
3028(3)
−510(1)
10959(1)
39(1)



C(5)
3801(3)
−104(1)
10329(1)
33(1)



C(6)
2600(3)
403(1)
9970(1)
37(1)



C(7)
3083(3)
859(1)
9363(1)
36(1)



C(8)
1807(3)
1350(2)
9016(1)
40(1)



C(9)
−379(3)
1525(2)
9198(1)
58(1)



C(10)
5961(3)
−331(1)
10158(1)
30(1)



C(11)
−833(4)
2464(2)
9220(1)
67(1)



C(12)
−225(4)
2915(2)
8572(1)
58(1)



C(13)
2015(3)
2770(1)
8411(1)
38(1)



C(14)
2368(3)
1800(1)
8383(1)
37(1)



C(15)
4498(3)
1722(1)
8100(1)
37(1)



C(16)
4621(3)
2444(1)
7585(1)
34(1)



C(17)
2724(3)
3012(1)
7691(1)
36(1)



C(18)
3346(4)
3155(1)
8960(1)
45(1)



C(20)
3039(4)
3933(1)
7453(1)
49(1)



C(20′)
4583(6)
4425(2)
7858(1)
72(1)



C(21)
3813(5)
3899(2)
6726(1)
57(1)



C(22A)
1282(8)
4571(3)
7457(3)
53(1)



C(23A)
−387(7)
4296(3)
6993(3)
71(2)



C(22B)
 749(8)
4221(5)
7469(4)
49(2)



C(23B)
 348(13)
5097(5)
7183(4)
71(3)

















TABLE 3





Bond lengths [Å] for 20DCM.


















O(1)—C(1)
1.426(2)



O(1)—H(1O)
0.8400



O(2)—C(3)
1.430(2)



O(2)—H(2O)
0.8400



C(1)—C(2)
1.510(2)



C(1)—C(10)
1.534(2)



C(1)—H(1)
1.0000



C(2)—C(2′)
1.321(3)



C(2)—C(3)
1.510(3)



C(2′)—H(2′A)
0.9500



C(2′)—H(2′B)
0.9500



C(3)—C(4)
1.524(3)



C(3)—H(3)
1.0000



C(4)—C(5)
1.507(3)



C(4)—H(4A)
0.9900



C(4)—H(4B)
0.9900



C(5)—C(6)
1.338(3)



C(5)—C(10)
1.512(3)



C(6)—C(7)
1.452(3)



C(6)—H(6)
0.9500



C(7)—C(8)
1.340(3)



C(7)—H(7)
0.9500



C(8)—C(14)
1.503(3)



C(8)—C(9)
1.516(3)



C(9)—C(11)
1.515(4)



C(9)—H(9A)
0.9900



C(9)—H(9B)
0.9900



C(10)—H(10A)
0.9900



C(10)—H(10B)
0.9900



C(11)—C(12)
1.535(4)



C(11)—H(11A)
0.9900



C(11)—H(11B)
0.9900



C(12)—C(13)
1.533(3)



C(12)—H(12A)
0.9900



C(12)—H(12B)
0.9900



C(13)—C(18)
1.535(3)



C(13)—C(14)
1.552(3)



C(13)—C(17)
1.565(3)



C(14)—C(15)
1.523(3)



C(14)—H(14)
1.0000



C(15)—C(16)
1.541(3)



C(15)—H(15A)
0.9900



C(15)—H(15B)
0.9900



C(16)—C(17)
1.556(3)



C(16)—H(16A)
0.9900



C(16)—H(16B)
0.9900



C(17)—C(20)
1.547(3)



C(17)—H(17)
1.0000



C(18)—H(18A)
0.9800



C(18)—H(18B)
0.9800



C(18)—H(18C)
0.9800



C(20)—C(20′)
1.518(4)



C(20)—C(22A)
1.537(4)



C(20)—C(21)
1.547(3)



C(20)—C(22B)
1.580(5)



C(20′)—H(20D)
0.9800



C(20′)—H(20E)
0.9800



C(20′)—H(20F)
0.9800



C(21)—H(21A)
0.9800



C(21)—H(21B)
0.9800



C(21)—H(21C)
0.9800



C(22A)—C(23A)
1.507(8)



C(22A)—H(22A)
0.9900



C(22A)—H(22B)
0.9900



C(23A)—H(23A)
0.9800



C(23A)—H(23B)
0.9800



C(23A)—H(23C)
0.9800



C(22B)—C(23B)
1.523(9)



C(22B)—H(22C)
0.9900



C(22B)—H(22D)
0.9900



C(23B)—H(23D)
0.9800



C(23B)—H(23E)
0.9800



C(23B)—H(23F)
0.9800

















TABLE 4





Bond angles [°] for 20DCM.


















C(1)—O(1)—H(1O)
109.5



C(3)—O(2)—H(2O)
109.5



O(1)—C(1)—C(2)
112.81(14)



O(1)—C(1)—C(10)
109.03(14)



C(2)—C(1)—C(10)
109.76(15)



O(1)—C(1)—H(1)
108.4



C(2)—C(1)—H(1)
108.4



C(10)—C(1)—H(1)
108.4



C(2′)—C(2)—C(3)
122.96(18)



C(2′)—C(2)—C(1)
123.77(18)



C(3)—C(2)—C(1)
113.25(15)



C(2)—C(2′)—H(2′A)
120.0



C(2)—C(2′)—H(2′B)
120.0



H(2′A)—C(2′)—H(2′B)
120.0



O(2)—C(3)—C(2)
106.43(15)



O(2)—C(3)—C(4)
112.18(16)



C(2)—C(3)—C(4)
110.58(17)



O(2)—C(3)—H(3)
109.2



C(2)—C(3)—H(3)
109.2



C(4)—C(3)—H(3)
109.2



C(5)—C(4)—C(3)
112.82(16)



C(5)—C(4)—H(4A)
109.0



C(3)—C(4)—H(4A)
109.0



C(5)—C(4)—H(4B)
109.0



C(3)—C(4)—H(4B)
109.0



H(4A)—C(4)—H(4B)
107.8



C(6)—C(5)—C(4)
120.34(18)



C(6)—C(5)—C(10)
125.47(18)



C(4)—C(5)—C(10)
114.19(18)



C(5)—C(6)—C(7)
128.22(18)



C(5)—C(6)—H(6)
115.9



C(7)—C(6)—H(6)
115.9



C(8)—C(7)—C(6)
125.78(18)



C(8)—C(7)—H(7)
117.1



C(6)—C(7)—H(7)
117.1



C(7)—C(8)—C(14)
123.88(17)



C(7)—C(8)—C(9)
125.5(2)



C(14)—C(8)—C(9)
110.64(19)



C(11)—C(9)—C(8)
112.0(2)



C(11)—C(9)—H(9A)
109.2



C(8)—C(9)—H(9A)
109.2



C(11)—C(9)—H(9B)
109.2



C(8)—C(9)—H(9B)
109.2



H(9A)—C(9)—H(9B)
107.9



C(5)—C(10)—C(1)
110.07(15)



C(5)—C(10)—H(10A)
109.6



C(1)—C(10)—H(10A)
109.6



C(5)—C(10)—H(10B)
109.6



C(1)—C(10)—H(10B)
109.6



H(10A)—C(10)—H(10B)
108.2



C(9)—C(11)—C(12)
112.3(2)



C(9)—C(11)—H(11A)
109.1



C(12)—C(11)—H(11A)
109.1



C(9)—C(11)—H(11B)
109.1



C(12)—C(11)—H(11B)
109.1



H(11A)—C(11)—H(11B)
107.9



C(13)—C(12)—C(11)
111.20(18)



C(13)—C(12)—H(12A)
109.4



C(11)—C(12)—H(12A)
109.4



C(13)—C(12)—H(12B)
109.4



C(11)—C(12)—H(12B)
109.4



H(12A)—C(12)—H(12B)
108.0



C(12)—C(13)—C(18)
110.03(17)



C(12)—C(13)—C(14)
107.5(2)



C(18)—C(13)—C(14)
109.37(15)



C(12)—C(13)—C(17)
116.55(16)



C(18)—C(13)—C(17)
113.17(19)



C(14)—C(13)—C(17)
99.38(15)



C(8)—C(14)—C(15)
120.36(17)



C(8)—C(14)—C(13)
113.59(16)



C(15)—C(14)—C(13)
103.42(17)



C(8)—C(14)—H(14)
106.2



C(15)—C(14)—H(14)
106.2



C(13)—C(14)—H(14)
106.2



C(14)—C(15)—C(16)
103.82(16)



C(14)—C(15)—H(15A)
111.0



C(16)—C(15)—H(15A)
111.0



C(14)—C(15)—H(15B)
111.0



C(16)—C(15)—H(15B)
111.0



H(15A)—C(15)—H(15B)
109.0



C(15)—C(16)—C(17)
107.05(16)



C(15)—C(16)—H(16A)
110.3



C(17)—C(16)—H(16A)
110.3



C(15)—C(16)—H(16B)
110.3



C(17)—C(16)—H(16B)
110.3



H(16A)—C(16)—H(16B)
108.6



C(20)—C(17)—C(16)
113.09(18)



C(20)—C(17)—C(13)
123.75(16)



C(16)—C(17)—C(13)
103.07(15)



C(20)—C(17)—H(17)
105.1



C(16)—C(17)—H(17)
105.1



C(13)—C(17)—H(17)
105.1



C(13)—C(18)—H(18A)
109.5



C(13)—C(18)—H(18B)
109.5



H(18A)—C(18)—H(18B)
109.5



C(13)—C(18)—H(18C)
109.5



H(18A)—C(18)—H(18C)
109.5



H(18B)—C(18)—H(18C)
109.5



C(20′)—C(20)—C(22A)
99.7(3)



C(20′)—C(20)—C(17)
114.01(17)



C(22A)—C(20)—C(17)
120.9(3)



C(20′)—C(20)—C(21)
107.5(2)



C(22A)—C(20)—C(21)
106.1(3)



C(17)—C(20)—C(21)
107.65(17)



C(20′)—C(20)—C(22B)
119.1(4)



C(17)—C(20)—C(22B)
97.8(3)



C(21)—C(20)—C(22B)
110.2(3)



C(20)—C(20′)—H(20D)
109.5



C(20)—C(20′)—H(20E)
109.5



H(20D)—C(20′)—H(20E)
109.5



C(20)—C(20′)—H(20F)
109.5



H(20D)—C(20′)—H(20F)
109.5



H(20E)—C(20′)—H(20F)
109.5



C(20)—C(21)—H(21A)
109.5



C(20)—C(21)—H(21B)
109.5



H(21A)—C(21)—H(21B)
109.5



C(20)—C(21)—H(21C)
109.5



H(21A)—C(21)—H(21C)
109.5



H(21B)—C(21)—H(21C)
109.5



C(23A)—C(22A)—C(20)
111.1(4)



C(23A)—C(22A)—H(22A)
109.4



C(20)—C(22A)—H(22A)
109.4



C(23A)—C(22A)—H(22B)
109.4



C(20)—C(22A)—H(22B)
109.4



H(22A)—C(22A)—H(22B)
108.0



C(23B)—C(22B)—C(20)
114.9(6)



C(23B)—C(22B)—H(22C)
108.5



C(20)—C(22B)—H(22C)
108.5



C(23B)—C(22B)—H(22D)
108.5



C(20)—C(22B)—H(22D)
108.5



H(22C)—C(22B)—H(22D)
107.5



C(22B)—C(23B)—H(23D)
109.5



C(22B)—C(23B)—H(23E)
109.5



H(23D)—C(23B)—H(23E)
109.5



C(22B)—C(23B)—H(23F)
109.5



H(23D)—C(23B)—H(23F)
109.5



H(23E)—C(23B)—H(23F)
109.5










Symmetry transformations used to generate equivalent atoms:








TABLE 5







Anisotropic displacement parameters (Å2 × 103) for 20DCM.


The anisotropic displacement factor exponent takes the form:


−2π2[h2a*2U11 + . . . + 2hka*b*U12]














U11
U22
U33
U23
U13
U12

















O(1)
17(1)
39(1)
35(1)
 7(1)
−2(1)
 1(1)


O(2)
19(1)
38(1)
49(1)
 3(1)
−5(1)
 1(1)


C(1)
16(1)
36(1)
25(1)
 5(1)
−2(1)
−2(1)


C(2)
23(1)
47(1)
28(1)
11(1)
−3(1)
−7(1)


C(2′)
29(1)
91(2)
41(1)
35(1)
−4(1)
−6(1)


C(3)
26(1)
49(1)
27(1)
 7(1)
 4(1)
−6(1)


C(4)
37(1)
49(1)
32(1)
−7(1)
11(1)
−7(1)


C(5)
30(1)
35(1)
34(1)
−7(1)
 9(1)
−5(1)


C(6)
27(1)
43(1)
42(1)
−7(1)
10(1)
 3(1)


C(7)
24(1)
45(1)
39(1)
−5(1)
 9(1)
 5(1)


C(8)
24(1)
59(1)
36(1)
−5(1)
 6(1)
 9(1)


C(9)
23(1)
109(2) 
43(1)
 5(1)
 8(1)
18(1)


C(10)
24(1)
35(1)
30(1)
 0(1)
 3(1)
−4(1)


C(11)
35(1)
124(3) 
43(1)
 7(1)
10(1)
47(2)


C(12)
37(1)
102(2) 
36(1)
 1(1)
 1(1)
43(1)


C(13)
31(1)
60(1)
23(1)
−6(1)
−4(1)
26(1)


C(14)
22(1)
58(1)
30(1)
−8(1)
 2(1)
10(1)


C(15)
30(1)
37(1)
43(1)
−6(1)
10(1)
10(1)


C(16)
29(1)
42(1)
30(1)
−7(1)
 2(1)
 6(1)


C(17)
34(1)
55(1)
21(1)
−10(1) 
−9(1)
19(1)


C(18)
59(1)
52(1)
23(1)
−7(1)
−10(1) 
30(1)


C(20)
69(2)
54(1)
22(1)
−5(1)
−10(1) 
33(1)


C(20′)
135(3) 
43(1)
37(1)
 3(1)
−22(2) 
 4(2)


C(21)
83(2)
61(1)
27(1)
−1(1)
−6(1)
25(2)


C(22A)
71(3)
32(2)
54(2)
 3(2)
 1(2)
26(2)


C(23A)
46(2)
60(3)
107(4) 
30(3)
−18(3) 
13(2)


C(22B)
72(4)
28(3)
47(3)
 5(3)
 1(3)
22(3)


C(23B)
70(5)
54(4)
88(5)
33(4)
 9(4)
20(4)
















TABLE 6







Hydrogen coordinates (×104) and isotropic displacement parameters


(Å2 × 103) for 20DCM.












x
y
z
U(eq)

















H(1O)
8320
−1979
9815
45



H(2O)
1091
−1624
10431
53



H(1)
5325
−1529
9785
31



H(2′A)
6255
−2317
11610
64



H(2′B)
8104
−2223
11066
64



H(3)
3079
−1679
11433
41



H(4A)
1559
−398
11000
47



H(4B)
3709
−248
11346
47



H(6)
1262
474
10134
44



H(7)
4422
805
9196
43



H(9A)
−1276
1251
8867
70



H(9B)
−672
1274
9640
70



H(10A)
6888
−84
10493
35



H(10B)
6316
−92
9717
35



H(11A)
−95
2722
9598
80



H(11B)
−2299
2547
9297
80



H(12A)
−1066
2702
8200
70



H(12B)
−482
3530
8618
70



H(14)
1435
1579
8031
44



H(15A)
4697
1165
7883
44



H(15B)
5528
1795
8453
44



H(16A)
4632
2212
7127
40



H(16B)
5872
2778
7653
40



H(17)
1668
2773
7388
44



H(18A)
3262
2804
9362
67



H(18B)
2873
3728
9062
67



H(18C)
4753
3179
8806
67



H(20D)
4016
4558
8297
108



H(20E)
4922
4952
7625
108



H(20F)
5809
4083
7913
108



H(21A)
5186
3665
6719
86



H(21B)
3829
4472
6538
86



H(21C)
2915
3539
6460
86



H(22A)
741
4623
7915
63



H(22B)
1788
5133
7318
63



H(23A)
−924
3749
7140
107



H(23B)
149
4242
6540
107



H(23C)
−1472
4719
6998
107



H(22C)
−64
3804
7216
59



H(22D)
274
4210
7937
59



H(23D)
595
5091
6701
106



H(23E)
1251
5509
7395
106



H(23F)
−1062
5256
7268
106

















TABLE 7





Torsion angles [deg] for 20DCM.


















O(1)—C(1)—C(2)—C(2′)
1.5(3)



C(10)—C(1)—C(2)—C(2′)
−120.3(2)



O(1)—C(1)—C(2)—C(3)
−179.83(15)



C(10)—C(1)—C(2)—C(3)
58.4(2)



C(2′)—C(2)—C(3)—O(2)
−113.9(2)



C(1)—C(2)—C(3)—O(2)
67.4(2)



C(2′)—C(2)—C(3)—C(4)
124.0(2)



C(1)—C(2)—C(3)—C(4)
−54.7(2)



O(2)—C(3)—C(4)—C(5)
−69.2(2)



C(2)—C(3)—C(4)—C(5)
49.5(2)



C(3)—C(4)—C(5)—C(6)
128.9(2)



C(3)—C(4)—C(5)—C(10)
−50.6(2)



C(4)—C(5)—C(6)—C(7)
178.79(18)



C(10)—C(5)—C(6)—C(7)
−1.8(3)



C(5)—C(6)—C(7)—C(8)
178.8(2)



C(6)—C(7)—C(8)—C(14)
−179.09(19)



C(6)—C(7)—C(8)—C(9)
0.6(4)



C(7)—C(8)—C(9)—C(11)
127.4(3)



C(14)—C(8)—C(9)—C(11)
−52.9(3)



C(6)—C(5)—C(10)—C(1)
−126.2(2)



C(4)—C(5)—C(10)—C(1)
53.3(2)



O(1)—C(1)—C(10)—C(5)
−179.82(14)



C(2)—C(1)—C(10)—C(5)
−55.79(19)



C(8)—C(9)—C(11)—C(12)
53.4(3)



C(9)—C(11)—C(12)—C(13)
−56.1(3)



C(11)—C(12)—C(13)—C(18)
−63.1(3)



C(11)—C(12)—C(13)—C(14)
56.0(3)



C(11)—C(12)—C(13)—C(17)
166.4(2)



C(7)—C(8)—C(14)—C(15)
−0.5(3)



C(9)—C(8)—C(14)—C(15)
179.8(2)



C(7)—C(8)—C(14)—C(13)
−123.9(2)



C(9)—C(8)—C(14)—C(13)
56.4(3)



C(12)—C(13)—C(14)—C(8)
−57.8(2)



C(18)—C(13)—C(14)—C(8)
61.7(2)



C(17)—C(13)—C(14)—C(8)
−179.57(16)



C(12)—C(13)—C(14)—C(15)
170.08(15)



C(18)—C(13)—C(14)—C(15)
−70.5(2)



C(17)—C(13)—C(14)—C(15)
48.27(18)



C(8)—C(14)—C(15)—C(16)
−165.00(19)



C(13)—C(14)—C(15)—C(16)
−36.93(18)



C(14)—C(15)—C(16)—C(17)
10.8(2)



C(15)—C(16)—C(17)—C(20)
154.93(16)



C(15)—C(16)—C(17)—C(13)
19.0(2)



C(12)—C(13)—C(17)—C(20)
74.9(3)



C(18)—C(13)—C(17)—C(20)
−54.2(2)



C(14)—C(13)—C(17)—C(20)
−170.08(19)



C(12)—C(13)—C(17)—C(16)
−155.4(2)



C(18)—C(13)—C(17)—C(16)
75.53(19)



C(14)—C(13)—C(17)—C(16)
−40.34(18)



C(16)—C(17)—C(20)—C(20′)
−66.7(3)



C(13)—C(17)—C(20)—C(20′)
58.8(3)



C(16)—C(17)—C(20)—C(22A)
174.4(3)



C(13)—C(17)—C(20)—C(22A)
−60.1(4)



C(16)—C(17)—C(20)—C(21)
52.4(2)



C(13)—C(17)—C(20)—C(21)
177.91(19)



C(16)—C(17)—C(20)—C(22B)
166.6(3)



C(13)—C(17)—C(20)—C(22B)
−67.9(4)



C(20′)—C(20)—C(22A)—C(23A)
171.3(4)



C(17)—C(20)—C(22A)—C(23A)
−62.9(5)



C(21)—C(20)—C(22A)—C(23A)
59.8(5)



C(22B)—C(20)—C(22A)—C(23A)
−43.7(9)



C(20′)—C(20)—C(22B)—C(23B)
63.7(7)



C(22A)—C(20)—C(22B)—C(23B)
23.3(8)



C(17)—C(20)—C(22B)—C(23B)
−173.2(6)



C(21)—C(20)—C(22B)—C(23B)
−61.1(7)

















TABLE 8







Hydrogen bonds for 20DCM [A and deg.]











D-H . . . A
d(D-H)
d(H . . . A)
d(D . . . A)
<(DHA)





O(1)—H(1O) . . . O(2)#1
0.84
2.03
2.8149(19)
154.8


O(2)—H(2O) . . . O(1)#2
0.84
2.08
2.8466(17)
150.9









Symmetry transformations used to generate equivalent atoms:

#1 x+1/2,−y−1/2,−z+2 #2 x−1,y,z

REFERENCES


    [1] CrysAlis RED, Oxford Diffraction Ltd.,Version 1.171.28cycle2 beta (release 25-10-2005 CrysAlis171.NET) (compiled Oct. 25, 2005, 08:50:05). Empirical absorption correction using spherical harmonics, implemented in SCALE3 ABSPACK scaling algorithm.
    [2] CrysAlis CCD, Oxford Diffraction Ltd., Version 1.171.28cycle2 beta; CrysAlis RED, Oxford Diffraction Ltd.,Version 1.171.28cycle2 beta
    [3] G. M. Sheldrick, Acta Crystallogr. 1990, A46, 467-473.
    [4] G. M. Sheldrick, SHELXL93. Program for the Refinement of Crystal Structures., Univ. of Göttingen, Germany.
    [5] International Tables for Crystallography, Ed. A. J. C. Wilson, Kluwer:Dordrecht, 1992, Vol.C.


EXAMPLE 4
Synthesis of 20DCM
The preparation of 20DCM having the basic structure I can be accomplished by a common general method, i.e. the condensation of a bicyclic Windaus-Grundmann type ketone II with the allylic phosphine oxide III to the corresponding 2-methylene-19-nor-vitamin D analog IV followed by deprotection at C-1 and C-3 in the latter compound IV to obtain compound I, i.e. 20DCM.




In phosphine oxide III, Y1 and Y2 are preferably hydroxy-protecting groups such as silyl protecting groups. The t-butyldimethylsilyl (TMDMS) group is an example of a particularly useful hydroxy-protecting group. The process described above represents an application of the convergent synthesis concept, which has been applied effectively to the preparation of numerous vitamin D compounds (see Lythgoe et al., J. Chem. Soc. Perkin Trans. I, 590 (1978); Lythgoe, Chem. Soc. Rev. 9, 449 (1983); Toh et al., J. Org. Chem. 48, 1414 (1983); Baggiolini et al., J. Org. Chem. 51, 3098 (1986); Sardina et al., J. Org. Chem. 51, 1264 (1986); J. Org. Chem. 51, 1269 (1986); DeLuca et al., U.S. Pat. No. 5,086,191; DeLuca et al., U.S. Pat. No. 5,536,713; and DeLuca et al, U.S. Pat. No. 5,843,928 all of which are hereby incorporated by reference in their entirety and for all purposes as if fully set forth herein.
Phosphine oxide III is a convenient reagent that can be used to prepare a large number of 19-nor-vitamin D compounds and is prepared according to the procedures described by Sicinski et al., J. Med. Chem., 41, 4662 (1998), DeLuca et al., U.S. Pat. No. 5,843,928; Perlman et al., Tetrahedron Lett. 32, 7663 (1991); and DeLuca et al., U.S. Pat. No. 5,086,191 which are hereby incorporated by reference in their entirety as if fully set forth herein.
The overall process of the synthesis of compound I is illustrated and described more completely in U.S. Pat. No. 5,843,928 entitled “2-Alkylidene-19-Nor-Vitamin D Compounds” and in application Ser. No. 12/343,602 filed Dec. 24, 2008, entitled “2-Methylene-20-Methyl-19,24,25,26,27-Pentanor-Vitamin D Analogs” published as U.S. Publication No. US 2009/0170822 the specifications of which are specifically incorporated herein by reference.

## Claims

1. 1α-hydroxy-20-methyl-2-methylene-19,24,25,26,27-pentanorvitamin D3 in crystalline form.

2. The crystalline form of 1α-hydroxy-20-methyl-2-methylene-19,24,25,26,27-pentanorvitamin D3 having molecular packing arrangement defined by space group P2 and unit cell dimensions a=6.6 Å b=15.8 Å c=20.0 Å α=90°, β=90° and γ=90°.

3. A three dimensional structure for 1α-hydroxy-20-methyl-2-methylene-19,24,25,26,27-pentanorvitamin D3 as defined by the molecular packing arrangement set forth in claim 2.

4. A method of purifying 1α-hydroxy-20-methyl-2-methylene-19,24,25,26,27-pentanorvitamin D3, comprising the steps of:
(a) preparing a solvent comprising ethyl formate;
(b) dissolving a product containing 1α-hydroxy-20-methyl-2-methylene-19,24,25,26,27-pentanorvitamin D3 to be purified in said solvent;
(c) cooling said solvent and dissolved product below ambient temperature for a sufficient amount of time to form a precipitate of 1α-hydroxy-20-methyl-2-methylene-19,24,25,26,27-pentanorvitamin D3 crystals; and
(d) separating the 1α-hydroxy-20-methyl-2-methylene-19,24,25,26,27-pentanorvitaminD3 crystals from the solvent.

5. The method of claim 4 including the further step of allowing said solvent and dissolved product to cool to ambient temperature prior to cooling below ambient temperature.

6. The method of claim 4 wherein said solvent comprises 100% ethyl formate, by volume.

7. The method of claim 4 wherein the step of separating comprises filtering the solvent and precipitate to obtain the crystals.

8. The method of claim 4 including a further step (e) comprising repeating steps (a) through (d) using the recovered crystals from step (d) as the product of step (b).

9. The method of claim 4 wherein said solvent comprises a mixture of ethyl formate and hexane.

10. The method of claim 9 wherein said mixture comprises about 75% ethyl formate and about 25% hexane, by volume.

11. A method of purifying 1α-hydroxy-20-methyl-2-methylene-19,24,25,26,27-pentanorvitamin D3, comprising the steps of:
(a) preparing a solvent comprising ethyl formate;
(b) dissolving a product containing 1α-hydroxy-20-methyl-2-methylene-19,24,25,26,27-pentanorvitamin D3 to be purified in said solvent;
(c) cooling said solvent and dissolved product below ambient temperature for a sufficient amount of time to form a precipitate of 1α-hydroxy-20-methyl-2-methylene-19,24,25,26,27-pentanorvitamin D3 crystals; and
(d) recovering the 1α-hydroxy-20-methyl-2-methylene-19,24,25,26,27-pentanorvitamin D3 crystals having a molecular packing arrangement defined by space group P2 and unit cell dimensions a=6.6 Å, b=15.8 Å, c=20.0 Å, α-90°, β=90° and γ=90°, or any other space group that yields substantially the same crystalline packing arrangement.

12. The method of claim 11 wherein said solvent and dissolved product is allowed to cool to ambient temperature prior to cooling below ambient temperature.

13. The method of claim 11 wherein said solvent comprises a mixture of ethyl formate and hexane.

14. The method of claim 13 wherein said mixture comprises about 75% ethyl formate and about 25% hexane, by volume.

15. The method of claim 11 wherein the step of recovering comprises filtering the solvent and precipitate to obtain the crystals.

16. The method of claim 11 including a further step (e) comprising repeating steps (a) through (d) using the recovered crystals from step (d) as the product of step (b).