CN1285573C - 3-Deoxy-vitamin D3 analogue ester - Google Patents

Abstract

The invention relates to compounds of formula or salts thereof, wherein the dotted line, R¹、R²、R³、R⁴And L is those groups defined in the claims. The invention also relates to compounds of formula or salts thereofUse as a medicament.

Description

3-Deoxy-vitamin D3 analogue ester

The present invention relates to methods of using vitamin _D3 analogs for the treatment of various diseases and methods of making these analogs. In particular, the present invention relates to 3-deoxy-20-demethyl-20-cyclopropyl vitamin D ₃ analog esters and methods of making and using these compounds.

Osteoporosis is the most common form of metabolic bone disease and can be considered a period of fracture with symptomatic bone loss (osteopenia). Although osteoporosis can be secondary to many underlying conditions, 90% of cases appear to be congenital. Postmenopausal women are at high risk for congenital osteoporosis (postmenopausal or type I osteoporosis); another group particularly at risk for osteoporosis is women of both sexes who have passed middle age (senile or type II osteoporosis). quality loose). Factors associated with osteoporosis also include: use of corticosteroids (corticosteroid hormones), immobilized or prolonged bed rest, alcoholism, diabetes mellitus, gonadotoxic chemotherapy, hyperprolactinemia, anorexia nervosa, primary and secondary amenorrhea, transplantation immunosuppression and oophorectomy. Postmenopausal osteoporosis is characterized by spinal fractures, whereas femoral neck fractures are a dominant feature of senile osteoporosis.

The bone loss mechanism is believed to involve a dysregulation of the bone's own regeneration process. This process is called bone remodeling. The bone regeneration process occurs in a series of dispersed active sacs. These cysts arise spontaneously in the bone matrix on the bone surface and serve as sites for bone resorption. Osteoclasts (cells that dissolve and resorb bone) are responsible for resorbing bone sections of generally constant size. This resorption process is followed by the appearance of osteoblasts (bone-forming cells), which fill the cavities left by osteoclasts with new bone cells.

In healthy adults, the function of osteoclasts and osteoblasts is to bring bone formation and bone resorption into balance. However, the imbalance in the bone remodeling process in patients with osteoporosis is enlarged, resulting in a rate of bone replacement that is lower than the rate of loss. Although in most individuals this dysregulation emerges to some extent with (years) aging, in postmenopausal osteoporotic patients undergoing oophorectomy, or in iatrogenic conditions, e.g. from The disorder is more severe and occurs earlier with corticosteroid therapy or with immunization practiced in organ transplantation.

Various methods have been suggested to increase bone mass in people suffering from osteoporosis, including administration of androgens, fluoride salts, and parathyroid hormone and modified parathyroid hormone. It has also been suggested that bisphosphonates, calcitonin, calcium, 1,25-dihydroxyvitamin _D3 and some of its analogs, and/or estrogens, alone or in combination, may be used to preserve existing bone quality.

Vitamin _D3 is a critical element in calcium metabolism, promoting intestinal absorption of calcium and phosphorus, maintaining adequate serum calcium and phosphorus concentrations, and stimulating calcium flow into and out of bone (tissue). Vitamin _D3 is hydroxylated in the body and the resulting 1α,25-dihydroxy metabolite is the active substance. Animal studies have shown that 1,25-(OH) ₂ vitamin _D3 has bone anabolic activity. Aerssens et al. in Calcif Tissue Int, 55:443-450 (1994) report the effect of 1α-hydroxyvitamin _D3 on bone strength and composition in corticosteroid-treated and corticosteroid-untreated growing rats. However, human use is limited to antiresorptives due to poor therapeutic ratios (hypercalciuria and hypercalcemia, and nephrotoxicity).

Dechant and Goa reported 1,25-di Hydroxyvitamin _D3 (calcitriol) has been shown to be effective in postmenopausal osteoporosis (promising for corticosteroid-induced osteoporosis), based on 622 postmenopausal patients with osteoporosis A clinical trial in women with porosis. Patients with mild to moderate disease (but not severe disease) who received calcitriol (0.25 μg twice daily) had significantly higher new The odds of a vertebral fracture were significantly reduced by 3 times. In patients starting long-term treatment with prednisone (prednisone) or prednisolone (prednisolone), calcitriol 0.5 to 1.0 μg per day plus 1000 mg of calcium with or without intranasal administration Calcitonin, 400 IU/day, prevented steroid-induced bone loss. In short, calcitriol has a good tolerance. At recommended doses, calcium intake and/or calcitriol doses are generally reduced accordingly, hypercalcemia is less frequent and symptoms are milder. However, calcitriol has a narrow therapeutic range, and its use requires adequate supervision and regular testing of serum (serum) calcium and creatinine concentrations. This study identifies a key limitation of calcitriol therapy, which lies in the close proximity of therapeutic and toxic doses.

Certain 3-deoxy-20-cyclopropyl vitamin D ₃ analogs have been disclosed to have cytostatic effects on prostate cancer cell lines in vitro (“20-Cyclopropyl-Cholecalciferol Vitamin D ₃ Analogs,” M. Koike et al., Anticancer Research , 19:1689-1698 (1999)).

Hyperparathyroidism

Secondary hyperparathyroidism is often found in patients with chronic renal failure. Decreased renal 1,25-(OH) ₂ vitamin _D3 (calcitriol) synthesis has been shown to be the main mechanism responsible for the secondary hyperparathyroidism in these patients, and calcitriol has been shown to have an effect on PTH synthesis. direct inhibition. Therefore, calcitriol is recommended for the treatment of secondary hyperparathyroidism in these patients. However, as noted above, calcitriol has a potentially hypercalcemic effect, making its therapeutic range narrow and limiting its use, especially at high doses. Accordingly, there is a need for an alternative method of treating hyperparathyroidism that does not produce these undesirable hypercalcemic effects.

Although a variety of compounds are available to treat these and other diseases, many of these compounds have undesirable side effects and/or are relatively unstable, ie, have a short shelf life. Therefore, there remains a need for additional compounds useful in the treatment of these diseases.

One aspect of the present invention provides a 3-deoxyvitamin _D3 analogue ester with the molecular formula:

or salts thereof, and methods of using and making such compounds, wherein

Dashed lines are optional double bonds;

L is a linking group selected from the following groups:

-CH ₂ -CH ₂ -CH ₂ -,

_-CH2 -CH=CH-,

-CH ₂ -C≡C-,

_-CH2 - _CH2 -C(=O)-, and

-CH=CH-CH=CH-;

R ² and R ³ each independently represent an alkyl or haloalkyl group; or R ² and R ³ together with the carbon atoms to which they are attached form a cycloalkyl group; and

R ^{and R} each independently represent hydrogen, alkyl, acyl or other hydroxyl protecting group,

The proviso is that at least one of ^R1 and ^R4 is an acyl group.

"Acetate" or "Ac" are used interchangeably herein to refer to a moiety of the formula -C(=O) _CH3 .

"Acyl" refers to a moiety of the formula -C(O)R', where R' is alkyl, heteroalkyl, cycloalkyl, aryl, heteroaryl, aralkyl, or heteroaralkyl.

"Alkyl" means a linear fully saturated hydrocarbon moiety containing 1 to 6, preferably 1 to 4 carbon atoms, or a branched fully saturated hydrocarbon moiety containing 3 or 6 carbon atoms.

"Aralkyl" refers to a moiety of the formula -Ra- ^Rb wherein ^Ra is alkyl as defined herein and ^Rb is aryl as defined ^herein .

"Aryl" means a monocyclic or bicyclic aromatic hydrocarbon moiety. Furthermore, one or more hydrogen atoms, preferably 1, 2 or 3 hydrogen atoms, on the aryl moiety may be substituted by halogen atoms, nitro, cyano, hydroxyl, amino, alkyl or alkoxy. Representative aryl groups include phenyl and naphthyl, which may be substituted with one or more of the substituents listed above. A preferred aryl group is phenyl.

"Cycloalkyl" refers to a fully saturated cyclic hydrocarbon moiety containing 3 to 6 ring carbon atoms, such as cyclopropyl, cyclopentyl and the like.

"Ester" means a compound containing a moiety of formula -O-C(=O)-R', where R' is alkyl, heteroalkyl, cycloalkyl, aryl, heteroaryl, aralkyl, or heteroaryl base.

"Haloalkyl" means an alkyl moiety as defined above in which one or more hydrogen atoms attached to the carbon backbone have been replaced by one or more halogen atoms. Preferred halides are fluorides.

"Heteroalkyl" means an alkyl moiety as defined herein containing one or more, preferably one, two or three substituents selected from -NR ^a R ^b , -OR ^c , wherein R ^a , R ^b and R ^c each independently represent a hydrogen atom, an alkyl group or a corresponding protecting group.

"Heteroaralkyl" refers to a moiety of the formula ^-Ra - ^Rb , wherein Ra ^{and Rb} are alkyl and heteroaryl, respectively, as defined herein.

"Heteroaryl" refers to a monocyclic or bicyclic group containing 5 to 12 ring-forming atoms, which contains at least one ring-forming heteroatom containing one, two or three ring-forming atoms selected from N, O or S, The remaining ring atoms are aromatic rings of C, with the understanding that the point of attachment of the heteroaryl group will be on the aromatic ring. Furthermore, one or more, preferably one, two or three hydrogen atoms of the heteroaryl moiety may be substituted by the abovementioned aryl substituents.

The terms "hydroxyl-protecting group" and "other hydroxy-protecting group" are used interchangeably herein to refer to hydroxy-protecting groups known to those skilled in the art other than alkyl or acyl groups, which are used in the present invention is specified. Representative hydroxy protecting groups include silyl ethers, carbonates, carbamates, substituted methyl ethers, substituted ethyl ethers, and the like. Lists of other suitable hydroxyl protecting groups can be found in, for example, Protective Groups in Organic Synthesis, Third Edition, T.W. Greene and P.G.M. Wuts, John Wiley & Sons, New York, 1999, which is incorporated herein by reference in its entirety.

"Pharmaceutically acceptable excipient" refers to an excipient that can be used in the preparation of a pharmaceutical composition, generally safe, non-toxic, and not biologically or otherwise undesirable. Excipients include veterinary and human excipients. Excipients for medication. A "pharmaceutically acceptable excipient" as used in the specification and claims includes one or more such excipients.

"Therapeutically effective amount" refers to the amount of the compound that is sufficient to produce therapeutic or preventive effects on the disease when administered to mammals. The "therapeutically effective amount" varies depending on the compound, the disease and its severity, and the age, weight, etc. of the patient being treated.

"Treatment" or "treatment" of disease includes: (1) prevention of disease, that is, the prevention of further development of clinical symptoms of disease in mammals that may be exposed to or susceptible to disease but have not yet experienced or shown symptoms of disease , (2) inhibiting the disease, that is, arresting or reducing the development of the disease or its clinical symptoms, or (3) alleviating the disease, that is, removing the disease or its clinical symptoms.

The terms "treating", "contacting" and "reacting" are used interchangeably herein when referring to a chemical reaction and refer to the addition or mixing of two or more reagents under appropriate conditions to produce the desired and/or or the desired product. It should be understood that the reaction leading to the desired and/or desired product may not necessarily result directly from the combination of the two reagents initially added, i.e., may be one or more intermediates formed in the mixture, ultimately forming the desired and/or desired product.

As used herein, the terms "defined above" and "herein defined" when referring to a variable incorporated by reference refer to a broad definition of the variable and, if there is a preferred range, to the preferred , more preferred and most preferred definitions.

"Haloalkyl" means an alkyl group as defined above wherein one or more hydrogen atoms attached to the carbon backbone have been replaced by one or more halogen atoms. Preferred halides are fluorides.

"Prodrug"refers to any compound that releases the active parent drug of formula (I) in vivo when such prodrug is administered to a mammal. The prodrug of the compound of formula (I) is prepared by modifying the functional group present in the compound of formula (I), and the modification can be decomposed in vivo to release the parent compound. Prodrugs include compounds of formula (I) wherein the hydroxyl group of compound (I) is bonded to any group that decomposes in vivo to yield a free hydroxyl group. Examples of prodrugs include, but are not limited to: esters (such as acetate, formate and benzoate derivatives), carbamates (such as N,N-dimethylamino carbonyl) and ethers of hydroxyl functional groups, etc. Conventionally, those skilled in the art acylate or etherify the hydroxyl groups in the parent compound to produce such compounds.

"Hydroxy protecting group" refers to a group of atoms which, when bound to a hydroxyl group in a molecule, shields, reduces or prevents the reactivity of the hydroxyl group. Examples of protecting groups can be found in T.W. Green and P.G. Futs, Protective Groups in Organic Chemistry, (Wiley, 2nd ed., 1999) and Harrison and Harrison et al., Compendium of Synthetic Organic Methods, vol. 1-8 (John Wiley and Sons, 1971-1996). Representative hydroxy protecting groups include groups in which the hydroxy group is acylated or alkylated, such as benzyl, as well as trityl ethers and alkyl ethers, tetrahydropyranyl ethers, trialkylsilyl ethers and alkenyl ethers. Propyl ether.

It has surprisingly been found that certain 3-deoxy-1α-hydroxy-20-cyclopropylcholecalciferol vitamin D ₃ compounds, the only ones known to date to have antiproliferative activity, are more effective than 1,25-dihydroxy Vitamin D ₃ is surprisingly effective at increasing bone formation. Accordingly, one aspect of the present invention provides a method of treating osteoporosis and hyperparathyroidism, the method comprising administering to a patient a 3-deoxy-1α-hydroxyvitamin D ₃ analogue, wherein the 3-deoxyvitamin D ₃ The structural formula of the analogue is as follows:

A prodrug or a salt thereof, preferably a compound of structural formula I or a salt thereof, wherein

Dashed lines are optional double bonds;

L is a linking group selected from the following groups:

-CH ₂ -CH ₂ -CH ₂ -,

_-CH2 -CH=CH-,

-CH ₂ -C≡C-,

_-CH2 - _CH2 -C(=O)-, and

-CH=CH-CH=CH-;

R ^{and R} are each selected from hydrogen or alkyl; and

R ² and R ³ are each independently selected from alkyl and haloalkyl, or R ² and R ³ together with the carbon atoms they are connected to form a cycloalkyl group.

Another aspect of the present invention provides a method of making a compound of the following structural formula:

The method involves converting a ketone having the following structural formula:

and a phosphine oxide compound of the formula:

contacting under conditions sufficient to produce said compound of formula I, wherein

Ar ^{and Ar} are each independently an optionally substituted aryl;

Dashed lines are optional double bonds;

L is a linking group selected from the following groups:

-CH ₂ -CH ₂ -CH ₂ -,

_-CH2 -CH=CH-,

-CH ₂ -C≡C-,

_-CH2 - _CH2 -C(=O)-, and

-CH=CH-CH=CH-;

R ^{and R} are each selected from hydrogen, alkyl and hydroxy protecting groups; and

R ² and R ³ are each independently selected from an alkyl group and a haloalkyl group; or R ² and R ³ form a cycloalkyl group together with the carbon atoms they are connected to.

In one embodiment of the process of the present invention for the manufacture of compounds of formula I, ^R4 is a hydrogen atom. In a specific embodiment, the method further comprises the step of protecting the hydroxyl group of the ketone before contacting the ketone of the formula II with the phosphine oxide compound of the formula III, and the ketone of the formula II and the phosphine oxide compound of the formula III The step of removing the hydroxy protecting group after contacting the phosphine oxide compound produces the compound of formula I.

Compounds of the following structural formula are preferred:

or a salt thereof, more preferably a compound of formula I,

in

Dashed lines are optional double bonds;

L is a linking group selected from the following groups:

-CH ₂ -CH ₂ -CH ₂ -,

_-CH2 -CH=CH-,

-CH ₂ -C≡C-,

_-CH2 - _CH2 -C(=O)-, and

-CH=CH-CH=CH-;

R ² and R ³ are each independently alkyl or haloalkyl; or R ² and R ³ together with the carbon atoms to which they are attached form a cycloalkyl group; and

R ^{and R} are each independently hydrogen, alkyl, acyl or other hydroxyl protecting groups,

The proviso is that at least one of ^R1 and ^R4 is an acyl group.

Also preferred are compounds of the formula:

in

R ¹ , R ² , R ³ , R ⁴ and L are those groups defined in claim 1 .

Another preferred aspect of the present invention are compounds of the following structural formula:

in

L is selected from the following groups:

-CH ₂ -CH ₂ -CH ₂ -;

_-CH2 -CH=CH-;

_-CH2 -C≡C-; and

-CH=CH-CH=CH-.

More preferred are compounds of formula I, wherein the linker L is selected from -CH ₂ -CH=CH-; and -CH ₂ -C≡C-.

Also preferred are compounds of formula I, wherein R ¹ is acyl.

Another preferred aspect of the invention are compounds of formula I, wherein R ⁴ is acyl.

Also preferred are compounds of formula I, wherein ^R2 and ^R3 are each independently selected from alkyl and haloalkyl.

Another preferred aspect of the invention are compounds of formula I, wherein ^R2 and ^R3 are trifluoromethyl.

Particular preference is given to compounds of formula I selected from the group consisting of:

[1R-(1α(E), 3aβ, 7aα)]-octahydro-7a-methyl-1-[5,5,5-trifluoro-4-hydroxy-4-(trifluoromethyl)-2- Pentynyl]cyclopropyl]-4H-inden-4-ol;

[1R-(1α(E), 3aβ, 7aα)]]-octahydro-7a-methyl-1-[1-[1-[5,5,5-trifluoro-4-hydroxyl-4-(tri Fluoromethyl)-2-pentynyl]cyclopropyl]-4H-inden-4-one;

[1R-[1α(E), 3aβ, 7aα)]]-octahydro-7a-methyl-1-[1-[5,5,5-trifluoro-4-trifluoromethyl)-4-[ (Trimethylsilyl)oxy]-2-pentynyl]cyclopropyl]-4H-inden-4-one;

3-deoxy-1,25-dihydroxy-20-methyl-23-(E)-ene-26,27-hexafluoro-21,28-cyclocholecalciferol;

3-deoxy-1,25-dihydroxy-20-methyl-23-(Z)-ene-26,27-hexafluoro-21,28-cyclocholecalciferol;

3-deoxy-1,25-dihydroxy-20-methyl-23-yne-21,28-cyclocholecalciferol;

3-deoxy-1,25-dihydroxy-20-methyl-23-yne-26,27-hexafluoro-21,28-cyclocholecalciferol;

3-deoxy-1,25-dihydroxy-20-methyl-21,28-cyclocholecalciferol;

3-deoxy-1α-acetoxy-25-hydroxy-20-cyclopropyl-23E-ene-26,27-hexafluoro-cholecalciferol; and

3-Deoxy-1α,25-diacetoxy-20-cyclopropyl-23E-ene-26,27-hexafluoro-cholecalciferol.

In addition, a method for producing a compound of structural formula I is also preferred, the method comprising:

(a) a ketone with the following structural formula:

and phosphine oxides of the formula:

contacting under conditions sufficient to produce said compound of formula I,

in

Ar ^{and Ar} are each independently an optionally substituted aryl;

Dashed lines are optional double bonds;

L is a linking group selected from the following groups:

-CH ₂ -CH ₂ -CH ₂ -,

_-CH2 -CH=CH-,

-CH ₂ -C≡C-,

_-CH2 - _CH2 -C(=O)-, and

-CH=CH-CH=CH-;

R ² and R ³ are each independently alkyl or haloalkyl; or R ² and R ³ together with the carbon atoms to which they are attached form a cycloalkyl group; and

R and R are ^each independently an alkyl, acyl or hydroxy ^protecting group; and

(b) when ^neither R ^nor R is an acyl group, acylation of a compound of formula I with an acylating agent under conditions sufficient to produce a compound of formula I, wherein at least one of R ^{and R} is acyl.

Particularly preferred is the above method wherein ^R1 and ^R4 are hydroxyl protecting groups.

Also particularly preferred is the above process, wherein the acylation step (b) comprises

(i) contacting the compound obtained in the step (a) and the hydroxy protecting group scavenging compound under conditions sufficient to produce a 1-hydroxyl-3-deoxyvitamin _D3 analogue of the following structural formula to remove the hydroxy group:

and

(ii) contacting a 1-hydroxy- ₃ -deoxyvitamin D3 analogue with an acylating agent under conditions sufficient to produce a 3-deoxyvitamin _D3 analogue ester of the formula

wherein R ^1a is acyl and R ^4a is hydrogen or acyl.

Furthermore, particularly preferred is the above method, wherein R ^4a is acyl.

Also preferred is the above method wherein Ar ¹ and Ar ² are phenyl.

Another preferred aspect of the present invention is the compound of formula I produced by the method described above.

Also preferred are pharmaceutical compositions comprising a compound of formula I and a pharmaceutically acceptable excipient.

More preference is given to using compounds of formula I as therapeutically active substances.

Another preferred aspect of the present invention is the compound of structural formula I for the preparation of a medicament for the prevention and treatment of bone-related diseases.

Preference is given to compounds of structural formula I for the preparation of medicaments for the prevention and treatment of hyperparathyroidism, renal osteodystrophy or osteoporosis.

Also preferred is a method of treating a bone-related disease in a patient comprising administering a compound of formula I to the patient.

More preferred is the method of formula I, wherein the disease is selected from hyperparathyroidism, secondary hyperparathyroidism, renal osteodystrophy and osteoporosis.

Another preferred aspect of the present invention is a method for treating bone-related diseases in a patient, the method comprising administering a compound of the following structural formula to the patient

Or a prodrug or a salt thereof, preferably a compound of structural formula I or a salt thereof, most preferably a compound of structural formula I,

in

Dashed lines are optional double bonds;

L is a linking group selected from the following groups:

-CH ₂ -CH ₂ -CH ₂ -,

_-CH2 -CH=CH-,

-CH ₂ -C≡C-,

_-CH2 - _CH2 -C(=O)-, and

-CH=CH-CH=CH-;

R ^{and R} are each selected from hydrogen or alkyl; and

R ² and R ³ are each independently selected from alkyl and haloalkyl, or R ² and R ³ together with the carbon atoms they are connected to form a cycloalkyl group. Particularly preferred is the above-mentioned method for treating bone-related diseases in a patient, the method comprising administering the compound of formula I to the patient.

It is also preferred to use compounds of the formula

Or prodrug or its salt, preferably structural formula I compound or its salt, most preferably structural formula I compound, prepare the medicine for the treatment and prevention of bone-related diseases,

in

Dashed lines are optional double bonds;

L is a linking group selected from the following groups:

-CH ₂ -CH ₂ -CH ₂ -,

_-CH2 -CH=CH-,

-CH ₂ -C≡C-,

_-CH2 - _CH2 -C(=O)-, and

-CH=CH-CH=CH-;

R ^{and R} are each selected from hydrogen or alkyl; and

R ² and R ³ are each independently selected from an alkyl group or a haloalkyl group, or R ² and R ³ form a cycloalkyl group together with the carbon atoms they are connected to.

Particularly preferred is the above use, wherein the bone-related disease is hyperparathyroidism, renal osteodystrophy or osteoporosis.

Preferably the above method or use, wherein the structural formula of the compound is as follows:

in

R ¹ , R ² , R ³ , R ⁴ and L are as defined above.

In addition, the above method or use is preferred, wherein the linker L is selected from:

-CH ₂ -CH ₂ -CH ₂ -;

_-CH2 -CH=CH-;

_-CH2 -C≡C-; and

-CH=CH-CH=CH-.

Also preferred is the above-mentioned method or the above-mentioned use, wherein the linker L is selected from:

_-CH2 -CH=CH-; and

_-CH2 -C≡C-.

More preferred is the above method or use, wherein R ¹ is hydrogen.

Also preferred is the above method or use, wherein ^R1 and ^R4 are hydrogen.

Particularly preferred is the above method or use, wherein ^R2 and ^R3 are each independently selected from alkyl and haloalkyl.

More particularly preferred is the aforementioned method or use, wherein ^R2 and ^R3 are both trifluoromethyl.

Furthermore, particularly preferred is the above method or use, wherein the disease is osteoporosis.

Additionally, preferred is the above method or use, wherein the disease is cured or the incidence of bone fractures is reduced.

Also particularly preferred is the above method or use, wherein the patient is at increased risk of fracture due to decreased bone mass.

Also preferred is the above method or use, wherein the patient is a female.

More preferred is the above method or use, wherein the patient has increased bone mineral density.

Also particularly preferred is a method or use as described above, wherein the patient is also treated with a bisphosphonate, an estrogen, a selective estrogen receptor modulator or an anabolic agent.

More preferred are methods of making compounds of the formula:

The method involves introducing a ketone of the formula:

and a phosphine oxide compound of the formula:

contacting under conditions sufficient to generate said compound of formula I,

in

Ar ^{and Ar} are each independently an optionally substituted aryl;

Dashed lines are optional double bonds;

L is a linking group selected from the following groups:

-CH ₂ -CH ₂ -CH ₂ -,

_-CH2 -CH=CH-,

-CH ₂ -C≡C-,

_-CH2 - _CH2 -C(=O)-, and

-CH=CH-CH=CH-;

R ^{and R} are each selected from hydrogen and alkyl; and

R ² and R ³ are each independently selected from alkyl and haloalkyl, or R ² and R ³ together with the carbon atoms they are connected to form a cycloalkyl group.

Particularly preferred is the above production method, wherein R ⁴ is hydrogen.

In addition, the above-mentioned production method is particularly preferred, which further includes the steps of: protecting the hydroxyl group of the ketone before the contact of the ketone of the structural formula II with the phosphine oxide compound of the structural formula III, and Removal of the hydroxy protecting group after contacting the ketone with the phosphine oxide compound of formula III yields the compound of formula I.

One aspect of the present invention provides a 3-deoxy-20-demethyl-20-cyclopropyl vitamin _D3 analog ester with the following structural formula:

or its salt,

in

Dashed lines are optional double bonds;

L is a linking group selected from the following groups:

-CH ₂ -CH ₂ -CH ₂ -,

_-CH2 -CH=CH-,

-CH ₂ -C≡C-,

_-CH2 - _CH2 -C(=O)-, and

-CH=CH-CH=CH-;

R ² and R ³ are each independently alkyl or haloalkyl; or R ² and R ³ together with the carbon atoms to which they are attached form a cycloalkyl group; and

^R1 and ^R4 are each independently hydrogen, alkyl, acyl or other hydroxyl protecting group, provided that at least one of ^R1 and ^R4 is acyl.

It has surprisingly been found that compounds of formula I in which at least one of R and ^R is an acyl group have unexpected stability and crystallinity compared to compounds in which ^{both R} and ^R are hydrogen, i.e. the parent diol good place.

When the cyclopentane ring part of formula I does not contain a double bond, ie when the dotted line does not exist, the stereochemical configuration of the side chain of the cyclopentane ring system can be α or β. The stereochemistry of the side chain of the preferred cyclopentane ring system is β type, that is, the following structural formula:

In a particularly preferred embodiment of the present invention, the structural formula of the ₃ -deoxyvitamin D analogue ester is as follows:

wherein R ¹ , R ² , R ³ , R ⁴ and L are those groups defined herein.

In yet another embodiment, the linker L is selected from the group consisting of _-CH2 - _CH2 - _CH2- ; _-CH2- CH=CH-; _-CH2- C≡C-; and -CH=CH-CH=CH -. Preferably L is -CH2 _- CH=CH- or _-CH2 -C≡C-. More preferably L is _-CH2 -CH=CH-, wherein the double bond is trans.

In another embodiment, it is preferred that ^R1 is acyl, more preferably acetyl.

In yet another embodiment, ^R1 is acyl and ^R4 is hydrogen or acyl.

In another embodiment, ^R1 is acyl, and ^R2 and ^R3 are each independently selected from methyl, ethyl and trifluoromethyl.

In yet another embodiment, ^R2 and ^R3 are alkyl or haloalkyl, preferably methyl or trifluoromethyl, most preferably trifluoromethyl.

The order of preference for the many different substituents given above and any of the substituent preferences below make compounds of the invention more preferred than compounds that do not follow a particular order of substituent preference. However, these substituent preferences are generally independent of each other, and although some preferences are mutually exclusive, the following multiple preferences may yield compounds that are more preferred than compounds that rarely follow substituent preferences.

The compound selected in the present invention

In a preferred aspect, the present invention relates to a method for treating patients with osteoporosis, hyperparathyroidism or autoimmune diseases, the medicine to be taken is a 3-deoxyvitamin D ₃ analogue of structural formula I, wherein R ¹ , R ² , R ³ , R ⁴ and L are as defined in the Summary of the Invention.

When the cyclopentane ring part of structural formula I does not contain a double bond, that is, when the dotted line does not exist or has no double bond, the stereochemical configuration of the side chain of the cyclopentane ring system can be α or β. The stereochemistry of the side chain of the preferred cyclopentane ring system is β type, that is, the following structural formula:

In one embodiment, the structural formula of the 3-deoxyvitamin _D3 analog is as follows:

wherein R ¹ , R ² , R ³ , R ⁴ and L are those groups defined in the Summary of the Invention.

In yet another embodiment, the linker L is selected from the group consisting of _-CH2 - _CH2 - _CH2- ; _-CH2- CH=CH-; _-CH2- C≡C-; and -CH=CH-CH=CH -. Preferably L is _-CH2 -CH=CH-; _-CH2 -C≡C-. More preferably L is _-CH2 -CH=CH-, wherein the double bond is trans.

In another embodiment, it is preferred that ^R1 is hydrogen.

In another embodiment, it is preferred that ^R4 is hydrogen.

In another embodiment, it is preferred that ^R1 and ^R4 are both hydrogen.

In yet another specific embodiment, ^R2 and ^R3 are each independently alkyl or haloalkyl, preferably methyl or trifluoromethyl.

The order of preference for the many different substituents given above and any of the substituent preferences below make compounds of the invention more preferred than compounds that do not follow a particular order of substituent preference. However, these substituent preferences are generally independent of each other, and although some preferences are mutually exclusive, the following multiple preferences may yield compounds that are more preferred than compounds that rarely follow substituent preferences.

General Synthetic Scheme

Although a variety of synthetic methods can be used to prepare compounds of formula I, one particular embodiment for preparing compounds of formula I involves coupling together a ketone and a phosphine oxide. Specifically, the compound of structural formula I can be prepared by the following reaction: the ketone of structural formula II:

and a phosphine oxide compound of the formula:

contacting under conditions sufficient to generate a compound of formula I,

in

Ar ^{and Ar} are each independently optionally substituted aryl;

Dashed lines are optional double bonds;

L is a linking group selected from the following groups:

-CH ₂ -CH ₂ -CH ₂ -,

_-CH2 -CH=CH-,

-CH ₂ -C≡C-,

_-CH2 - _CH2 -C(=O)-, and

-CH=CH-CH=CH-;

R ² and R ³ are each independently alkyl or haloalkyl; or R ² and R ³ together with the carbon atoms to which they are attached form a cycloalkyl group; and

R and R are ^each independently an alkyl, acyl or ^hydroxy protecting group, and

(b) when ^neither R ^nor R is an acyl group, acylation of the compound of formula I with an acylating agent under conditions sufficient to produce a compound of formula I wherein at least one of R ^{and R} is acyl.

In one embodiment, ^R1 and ^R4 are hydroxyl protecting groups. In this particular embodiment, the acylation step (b) comprises:

(i) Under conditions sufficient to produce 1-hydroxyl-3-deoxyvitamin _D3 analogues of the following structural formula, the compound obtained in step (a) is contacted with a hydroxyl protecting group scavenging compound to remove the hydroxyl protecting group:

and

(ii) reacting 1-hydroxy-3 _- deoxyvitamin D3 analogs and an acylating agent under conditions sufficient to produce a 3-deoxyvitamin _D3 analog ester of the following structural formula:

wherein R ^1a is acyl and R ^4a is hydrogen or acyl.

In another embodiment, R ^4a is acyl.

In yet another embodiment ^Ar1 and ^Ar2 are phenyl.

Suitable hydroxy protecting groups are well known to those of ordinary skill in the art and examples of such hydroxy protecting groups are disclosed in: Protective Groups in Organic Synthesis, Third Edition, T.W. Greene and P.G.M. Wuts, John Wiley & Sons, New York , 1999, and Harrison and Harrison et al., Compendium of Synthetic Organic Methods, Volumes 1-8 (John Wiley and Sons, 1971-1996), the entire contents of which are incorporated herein by reference. Representative hydroxy protecting groups include benzyl and trityl ethers, tetrahydropyranyl ethers, trialkylsilyl ethers, carbamates and allyl ethers.

Typically, the hydroxyl group is protected as a silyl ether; however, the scope of the present invention includes the use of alternative hydroxyl protecting groups known in the art, as disclosed above in Protective Groups in Organic Synthesis, Third Edition, and Compendium of Protecting groups as described in Synthetic Organic Methods, Volumes 1-8.

Typically, the phosphine oxide of formula III in tetrahydrofuran is reacted with n-butyllithium, typically at about -78°C. A solution of the ketone of formula II in tetrahydrofuran is then added to the mixture to prepare the compound of formula I. As specified above, when ^R1 and/or ^R4 are hydrogen, they are protected with a hydroxy protecting group prior to the coupling reaction, in which case the hydroxy protecting group is removed to give the compound of formula I.

Synthesis and purification of compounds of formula III are known and routine in the art. See, for example, M.R. Uskokovic et al., "Vitamin D Gene Regulation, Structure Function Analysis and Clinical Application," Paris, France, pp. 139-145 (1991), U.S.P. 5,086,191 and 5,616,759 by DeLuca et al., 5,087,619 by Baggiolini et al. 5,384,314 of Doran et al., 5,428,029 of Doran et al., 5,451,574 of Baggiolini et al.; European Patent Publication EP 0 808,832A2, patent publication WO 96/31216 of Brasitus et al.; Shiuey et al., J.Org.Chem., 55:243-247 (1990 ), Kiegel, J. et al. and Tetr. Lett., 32:6057-6060 (1991), Perlman, K.L. et al., Tetr. Lett., 32:7663-7666 (1991).

Reaction Scheme I illustrates a synthetic method for preparing compounds of formula IA.

            Plan I

In reaction scheme I, the compound of structural formula IV is a known compound, is prepared by WO99/12894 (1,3-dihydroxy-20,20-dialkylvitamin D3 analogue published on March 18, 1999) ) prepared by the method described in ). Compounds of formula IV are converted to compounds of formula V by selective partial reduction of the triple bond to the anb E-double bond with lithium aluminum hydride in an inert organic acid such as tetrahydrofuran. The reaction is generally carried out by adding the compound of structural formula IV to a suspension of LiAlH ₄ in THF at 0°C or 5°C. Heating the reaction mixture under reflux conditions affords the compound of formula V. Compounds of formula V are converted to ketones of formula VI using an oxidizing agent, such as pyridinium dichromate. This step is generally carried out in a halogenated solvent such as dichloromethane at room temperature. A silylating agent such as 1-(trimethylsilyl)imidazole, trimethylsilyl chloride or trimethylsilyl trifluoride (triflate) is then used in an inert solvent such as a halogenated solvent such as dichloro methane), the hydroxyl group of the compound of formula VI is protected in the form of a silyl ether of formula VII at room temperature. The compound of structural formula IIIA is reacted with n-butyllithium, and the compound obtained is reacted with the compound of structural formula VII in tetrahydrofuran. The reaction is generally carried out at a temperature of about -78° C., and then the silyl protecting group is removed, for example, tetrahydrofuran is used in tetrahydrofuran solvent. Removal of the silyl protecting group with ammonium fluoride affords compounds of formula IA'. The free hydroxyl group is then acetylated, eg, with acetic anhydride in pyridine, to give compounds of formula IA. Since secondary hydroxyl groups are generally more reactive, selective acetylation can be performed depending on the amount of acetylating reagent used and/or reaction conditions, such as reaction temperature and/or reaction time. Alternatively, both secondary and tertiary hydroxyl groups can be acetylated using excess acetylating reagent and prolonged reaction time.

Similarly, Z-stereoisomer analogs or saturated carbon chain analogs of compounds of formula IA can be prepared respectively by the reduction of compounds of formula IV with hydrogen in the presence of a suitable hydrogenation catalyst, such as Pd—S or Pd . The corresponding Z-isomer analogs or saturated carbon chain analogs of the compound of formula IA can be produced from the resulting compound as shown in Scheme I under similar reaction conditions.

As shown in Reaction Scheme II, compounds of formula II containing acetylenic alcohols and variable alkyl, haloalkyl, and cycloalkyl groups of R ^{and R} can be prepared by reacting compounds derived from compounds of formula VIII (wherein Pg is hydroxy Protecting group) is condensed with an appropriate ketone, haloketone (eg hexafluoroacetone) and cyclic ketone to remove the protecting group.

Scheme II

The compound of formula X is then reacted (ie, oxidized, protected and coupled) under similar reaction conditions as shown in Reaction Scheme I above to produce a compound of formula I containing an acetylenic linker moiety.

In certain preferred embodiments, the dashed line is a double bond, i.e. a compound of the formula:

In another preferred embodiment, the linking group L is selected from the following groups:

-CH ₂ -CH ₂ -CH ₂ -,

_-CH2 -CH=CH-,

_-CH2 -C≡C-, and

-CH=CH-CH=CH-.

Preferably R ¹ is acyl.

Preferably ^R4 is hydrogen or acyl.

Preferably, R ² and R ³ are each independently selected from methyl, ethyl and trifluoromethyl, or R ² and R ³ form a cyclopentyl ring together with the carbon atoms to which they are attached.

The order of preference for the many different substituents given above and any of the substituent preferences below make compounds of the invention more preferred than compounds that do not follow a particular order of substituent preference. However, these substituent preferences are generally independent of each other, and although some preferences are mutually exclusive, the following multiple preferences may yield compounds that are more preferred than compounds that rarely follow substituent preferences.

Another aspect of the invention provides salts of compounds of formula I.

Although a variety of synthetic methods can be used to prepare compounds of formula I, a specific embodiment for preparing compounds of formula I is illustrated below.

The compound of structural formula I can be prepared by following reaction, the compound of structural formula II

React with a phosphine oxide of the following structural formula,

wherein Ar ¹ and Ar ² are independently optionally substituted aryl groups, the meanings of the dashed lines, R ¹ , R ² , R ³ , R ⁴ and L are as defined in the Summary of the Invention. Preferably Ar ¹ and Ar ² are phenyl. When R ¹ and/or R ⁴ are hydrogen, it is preferable to protect the corresponding hydroxyl group with a hydroxyl protecting group matching the coupling reaction conditions before the coupling reaction between the ketone of formula II and the phosphine oxide compound of formula III. Suitable hydroxy protecting groups are well known to those of ordinary skill in the art and examples of such hydroxy protecting groups are disclosed in: Protective Groups in Organic Synthesis, Third Edition, TW Greene and PGM Wuts, John Wiley & Sons, New York, 1999 , and Harrison and Harrison et al., Compendium of Synthetic Organic Methods, Vol. 1-8 (John Wiley and Sons, 1971-1996), the entire contents of which are incorporated herein by reference. Representative hydroxy protecting groups include benzyl and trityl ethers, tetrahydropyranyl ethers, trialkylsilyl ethers and allyl ethers.

Typically, the hydroxyl group is protected as a silyl ether; however, the scope of the present invention includes the use of alternative hydroxyl protecting groups known in the art, as disclosed above in Protective Groups in Organic Synthesis, Third Edition, and Compendium of Protecting groups as described in Synthetic Organic Methods, Volumes 1-8.

Typically, the phosphine oxide compound of formula III in tetrahydrofuran is reacted with n-butyllithium, typically at about -78°C. A solution of the ketone of formula II in tetrahydrofuran is then added to the mixture to prepare the compound of formula I. As mentioned above, when ^R1 and/or ^R4 are hydrogen, they are protected with a hydroxyl protecting group prior to the coupling reaction. In this case, the hydroxy protecting group is then removed to obtain the compound of formula I.

Synthesis and purification of compounds of formula III are known and routine in the art. See, for example, M.R. Uskokovic et al., "Vitamin D Gene Regulation, Structure Function Analysis and Clinical Application," Paris, France, pp. 139-145 (1991), U.S.P. 5,086,191 and 5,616,759 by DeLuca et al., 5,087,619 by Baggiolini et al. 5,384,314 of Doran et al., 5,428,029 of Doran et al., 5,451,574 of Baggiolini et al.; European patent publication EP 0 808,832A2, patent publication WO 96/31216 of Brasitus et al.; Shiuey et al., J.Org.Chem., 55:243-247 (1990 ), Kiegel, J. et al., Tetr. Lett., 32:6057-6060 (1991), Perlman, K.L. et al., Tetr. Lett., 32:7663-7666 (1991).

Reaction Scheme Ia illustrates a synthetic method for preparing compounds of formula IA.

Option Ia

In reaction scheme Ia, the compound of structural formula IV is a known compound, is prepared by WO99/12894 (1,3-dihydroxy-20,20-dialkylvitamin D3 analogue published on March 18, 1999) ) prepared by the method described in ). Compounds of formula IV are converted to compounds of formula V by selective partial reduction of the triple bond to the E-double bond with lithium aluminum hydride in an inert organic acid such as tetrahydrofuran. The reaction is generally carried out by adding the compound of structural formula IV to a suspension of LiAlH ₄ in THF at 0°C or 5°C. Heating the reaction mixture under reflux conditions affords the compound of formula V. Compounds of formula V are oxidized to ketones of formula VI with an oxidizing agent, such as pyridinium dichromate. This step is generally carried out at room temperature in a halogenated solvent such as dichloromethane. A silylating agent such as 1-(trimethylsilyl)imidazole, trimethylsilyl chloride or trimethylsilyl trifluoride is then used in an inert solvent such as a halogenated solvent such as dichloromethane , protecting the hydroxyl group of the compound of the formula VI in the form of a silyl ether of the formula VII at room temperature. The compound of structural formula IIIa is reacted with n-butyllithium, and the compound obtained is reacted with the compound of structural formula VII in tetrahydrofuran at a reaction temperature of -78°C, and then the silyl protecting group is removed, for example, in tetrahydrofuran solvent with tetrabutylammonium fluoride. A silyl protecting group affords compounds of formula IA.

Similarly, Z-stereoisomer analogs or saturated carbon chain analogs of compounds of formula IA can be prepared respectively by the reduction of compounds of formula IV with hydrogen in the presence of a suitable hydrogenation catalyst, such as Pd—S or Pd . The corresponding Z-isomer analogs and saturated carbon chain analogs of the compound of formula IA can be prepared from the resulting compound under similar reaction conditions as shown in Scheme Ia.

As shown in Reaction Scheme 2a, compounds of formula II containing acetylene alcohols and variable alkyl, haloalkyl and cycloalkyl groups of R ^{and R} can be prepared by reacting compounds derived from compounds of formula VIII (wherein Pg is The protecting group is removed by condensation of the acetylenated anion of the hydroxy protecting group) with the appropriate ketone, haloketone (eg hexafluoroacetone) and cyclic ketone.

Option 2a

Compounds of formula X are then reacted (ie, oxidized, protected and coupled) under similar reaction conditions as shown in Reaction Scheme Ia above to produce compounds of formula I containing an acetylenic linker moiety.

Practicality

The compounds of the present invention are useful in the prevention and treatment of a variety of mammalian diseases in which bone loss is manifested. All of these diseases are referred to as "bone-related diseases" and are described in more detail below. In particular, the compounds of the invention have been shown to prevent and treat osteoporosis and osteopenia in mammals without inducing hypercalciuria, hypercalcemia or nephrotoxicity. "Hypercalcemia" is an excessively high concentration of calcium in the blood serum; the corresponding concentration in humans (and rats) is greater than about 10.5 mg/dl. "Intolerable hypercalcemia" usually occurs at serum calcium concentrations greater than about 12 mg/dl and is associated with emotional lability, confusion, confusion, psychosis, stupor, and coma.

The compounds of the present invention are useful in the treatment of type I (postmenopausal), type II (iatrogenic) and type III (senile) osteoporosis, including diseases associated with corticosteroid therapy (e.g. treatment of asthma), It can also treat osteodystrophy due to kidney dialysis and hyperparathyroidism. Treatment with the vitamin D3 analogues described herein results in increased bone mineral density and provides high quality bone mass unlike conventional treatments. Thus, the treatments described here may reduce the incidence of fractures and allow more rapid healing of existing fractures. This treatment is particularly useful in patients suffering from estrogen decline, such as older women, who would be at increased risk of fractures with other treatments. Treatable fracture types include traumatic and osteoporotic fractures such as hip, femoral neck, wrist, vertebrae, spine, ribs, sternum, larynx and trachea, radius/ulna, tibia, patella, clavicle, pelvis, humerus, Fractures of lower legs, fingers and toes, face and ankles.

The compounds of the present invention are also useful in the treatment of diseases caused by elevated concentrations of parathyroid hormone. In one aspect, the compounds of the invention are used to treat secondary hyperparathyroidism associated with renal failure, in particular to reverse or reduce bone loss associated with renal insufficiency. Other aspects include the treatment of renal osteodystrophy associated with advanced secondary hyperparathyroidism. Other aspects include the treatment of primary hyperparathyroidism.

Compounds of formula I are also useful in the treatment of neoplastic diseases such as leukemia, colon cancer, breast cancer and prostate cancer.

In general, the compounds of the present invention do not cause the elevated calcium concentrations observed with other vitamin _D3 analogues, such as 1,25(OH) _{2vitamin D3} , thus increasing the rate of treatment and better treating the aforementioned disease.

Administration and pharmaceutical composition

Generally, the administration amount of the compound of the present invention may be about 0.0002 to 0.5 mg compound/kg body weight per day, preferably about 0.001 to about 0.1 mg/kg body weight per day, more preferably about 0.002 to about 0.02 mg/kg body weight per day, most preferably Preferably from about 0.005 to about 0.010 mg/kg body weight per day. For a 50 kg human subject, the daily dose of active ingredient may be from about 0.01 to about 25 mg, preferably from about 0.05 to about 10 mg, most preferably from about 1.0 to about 10 mg per day. The dose can be in the form of a conventional pharmaceutical composition, according to the need to obtain the best effect, through single administration, combined administration or controlled release, preferably once or twice a day orally. In some instances, varying the daily dosage may be appropriate to achieve the desired therapeutic response.

The selection of the exact dosage and composition as well as the optimal route of administration is influenced by, inter alia, the pharmacological properties of the formulation, the nature and severity of the disease to be treated, and the physical condition and mental acuity of the excipients . In general, higher doses of corticosteroid are necessary for the treatment of corticosteroid-induced osteopenia.

Representative routes of administration include oral, parenteral (including subcutaneous, intramuscular, and intravenous), rectal, buccal (sublingual), pulmonary, transdermal, and nasal Internal administration, most preferably orally. Administration can be continuous or intermittent (eg, by bolus injection).

A related aspect of the invention relates to combination therapy of a compound of formula I and other active agents, such as bisphosphonates, estrogens, SERMS (selective estrogen receptor modulators), calcitonin or anabolic therapy. Examples of bisphosphonates include alendronate, ibandronate, pamidronate, etidronate and risedronate. Examples of SERMS include raloxifene, dihydroraloxifene and lasofoxifene. Calcitonin includes human and salmon calcitonin. Anabolic agents include parathyroid hormone (PTH), such as hPTH(1-34), PTH(1-84), and parathyroid hormone-related protein (PTHrP) and analogs thereof. Specific PTHrP analogs are described in the following documents: "Mono-and Bicyclic Analogs of Parathyroid Hormone-Related Protein. 1. Synthesis and Biological Studies," Michael Chorev et al., Biochemistry, 36:3293-3299 (1997) and "Cyclic analogs of PTH and PTHrP," WO 96/40 193 and U.S.P 5,589,452 and WO 97/07815. The other active agent can be used simultaneously, before or after the use of the compound of formula I, and different modes of administration can be used.

Another aspect of the present invention relates to a pharmaceutical composition comprising a compound of the present invention as an active ingredient in admixture with a pharmaceutically acceptable non-toxic carrier. As mentioned above, this composition can be prepared in the form of parenteral administration (subcutaneous administration, intramuscular injection and intravenous injection), especially in the form of liquid solution or suspension; oral or buccal administration, especially Tablet or capsule form; Pulmonary or intranasal form, especially powder, nose drops or aerosol form; and Rectal or transdermal form.

The compositions of the present invention may conveniently be administered in unit dosage form and may be prepared by any of the methods well known in the pharmaceutical art, e.g., as described in Remington's Pharmaceutical Sciences, 17th Edition, Mack Publishing Company, Easton, PA (1985). method. Formulations for parenteral administration may contain the following as excipients: sterile water or saline; alkylene glycols such as propylene glycol; polyalkylene glycols such as polyethylene glycol; vegetable oils; hydrogenated naphthalene etc. Formulations for nasal administration may be in solid form, may contain excipients such as lactose or dextran, or may be aqueous or oily solutions for use in the form of nasal drops or metered sprays. For buccal administration typical excipients include sugar, calcium stearate, magnesium stearate, pregelatinized starch, and the like.

Orally administrable compositions may include one or more physiologically compatible carriers and/or excipients, and may be in solid or liquid form. Tablets and capsules can be prepared with binders such as syrup, gum arabic, gelatin, sorbitol, tragacanth, or polyvinylpyrrolidone; fillers such as lactose, sucrose, corn starch, calcium phosphate, sorbitol; or glycine; lubricants, such as magnesium stearate, talc, polyethylene glycol, or silicon dioxide; and surfactants, such as sodium lauryl sulfate. Liquid compositions may contain the following substances: conventional additives such as suspending agents such as sorbitol syrup, methylcellulose, sugar syrup, gelatin, carboxymethylcellulose, or edible fats; emulsifiers such as lecithin or acacia vegetable oils such as almond oil, coconut oil, cod liver oil, or peanut oil; preservatives such as butylated hydroxyanisole (BHA) and butylated hydroxytoluene (BHT). Liquid compositions are enclosed in, for example, gelatin capsules to form unit dosage forms.

Preferred solid oral dosage forms include tablets, two-piece hard shell capsules and soft elastic gelatin (SEG) capsules. SEG capsules are of particular interest because of their clear advantages over the other two forms (see Seager, H., "Soft gelatin capsules: a solution to any tableting problems"; Pharmaceutical Technology, 9, (1985)). Some of the advantages of using SEG capsules are: a) Dose uniformity is optimized in SEG capsules because the drug is dissolved or dispersed in a liquid and can be precisely dosed into the capsule, b) Drugs formulated into SEG capsules perform well bioavailability because the drug is dissolved, solubilized, or dispersed in a water-miscible or oily liquid, so that when released in vivo, the solution dissolves or is emulsified to form a high surface area drug dispersion, and c) prevents long-term Degradation of easily oxidized drugs during storage because the soft gelatin dry shell forms a barrier against oxygen diffusion.

Dry shell formulations generally contain gelatin at a concentration of about 40%-60%, plasticizers (such as glycerin, sorbitol, or propylene glycol) at a concentration of about 20%-30%, and water at a concentration of about 30-40%. Other materials such as preservatives, dyes, opacifiers and fragrances may also be present. Liquid-fill materials include solid drugs that have been dissolved, solubilized, or dispersed (with suspending agents such as beeswax, hydrogenated castor oil, or polyethylene glycol 4000), or liquid drugs in a carrier or combination of carriers, such as mineral oil , vegetable oils, triglycerides, glycols, polyols and surfactants.

The following examples are given to enable those skilled in the art to understand and practice the present invention more clearly. These examples should not be considered limitations on the scope of the invention, but merely illustrative and representative examples.

Example

The following examples are intended to illustrate rather than limit the invention claimed.

Example 1

This example illustrates that [1R-(1α(E),3aβ,7aα)]-octahydro-7a-methyl-1-[5,5,5-trifluoro-4-hydroxy-4-(trifluoromethyl The preparation method of -2-pentynyl] cyclopropyl] -4H-inden-4-ol.

To a stirred, chilled (5° C.) suspension of LiAlH ₄ (237.2 mg; 6.25 mmol) in anhydrous THF (6.0 ml) was added NaOMe (338 mg, 6.25 mmol) powder. The mixture was stirred at 5°C under Ar for 15 min, and treated with [1R-(1α,3aβ,4α,7aα)]-octahydro-7a-methyl-1-[1-[5,5,5-trifluoro-4 -Hydroxy-4-(trifluoromethyl)-2-pentynyl]cyclopropyl]-4H-inden-4-ol (500mg, 1.25mmol) in anhydrous THF (6.0ml) was treated at reflux Boiling for 2.5 hours. After cooling, the mixture was diluted with _Et2O (25ml), quenched with dropwise addition of water (2.0ml) and 2M NaOH (2.0ml), and stirred at room temperature for 30 minutes. _MgSO4 (5g) was added and stirred for a further 30 minutes, then the mixture was diluted with _Et2O (25ml) and filtered through celite (15g) with ethyl acetate (3 x 20ml). Evaporation gave a gum (508 mg) which was purified by flash chromatography (50 g silica gel, 3.5 cm diameter column, 30% ethyl acetate in hexanes) in 20 ml fractions. Fractions 7-12 were evaporated to give colorless crystals (486mg), which were triturated with hexane and collected by filtration to give the title compound (442mg, 88%): mp 122-123°C; [α] _D +42.1° (ethanol, c= 0.80); IR 3540, 1602, 965cm ^-1 ; ¹ H NMR δ0.05(1H, m), 0.27(1H, m), 0.34(1H, m), 0.74(1H, m), 0.98(1H, m ), 1.00(3H, s), 1.17-1.25(2H, m), 1.35-1.60(6H, m), 1.65(1H, dd, J=12, 5), 1.75-1.87(3H, m), 2.02 (1H, d, J=11), 2.78 (1H, dd, J=14, 8.5), 2.92 (1H, s, OH), 4.05 (1H, br s), 5.59 (1H, d, J=16) , 6.30 (1H, ddd, J=16, 8.5, 6); MS m/z 400 (M ⁺ , 10). Anal. Calcd. for C ₁₉ H ₂₆ F ₆ O ₂ : C, 56.99; H, 6.55; F, 28.47 . Measured: C, 56.87; H, 6.33; F, 28.69.

Example 2

This example illustrates that [1R-(1α(E),3aβ,7aα)]]-octahydro-7a-methyl-1-[1-[1-[5,5,5-trifluoro-4-hydroxyl -4-(trifluoromethyl)-2-pentynyl]cyclopropyl]-4H-inden-4-one.

To the stirred [1R-(1α(E),3aβ,4α,7aα)]-octahydro-7a-methyl-1-[5,5,5-trifluoro-4-hydroxy-4-(trifluoro To a solution of methyl)-2-pentynyl]cyclopropyl]-4H-indene-4-ol (400mg, 1.00mmol) in CH ₂ Cl ₂ (8.0ml) was added ground pyridinium dichromate (1.25 g, 3.3 mmol). The mixture was stirred at room temperature for 4.5 hours, diluted with diisopropyl ether (15ml) and worked up to give 395mg of a colorless gum. Flash chromatography (50 g silica gel, 3.5 cm diameter column, 30% ethyl acetate in hexane) chromatography, each 20 ml was collected in fractions, fractions 7-12 were evaporated to give the title compound as colorless crystals (376 mg, 94% ): mp 79-80°C; [α] _D +6.9° (ethanol, c=1.00); IR 3334, 1704, 964cm ^-1 ; 1H NMR δ0.14(1H, m), 0.33(1H, m), 0.69 (1H, m), 0.70 (3H, s), 1.46-1.80 (5H, m), 1.91-2.30 (6H, m), 2.44 (1H, dd, J=11, 6), 2.74 (1H, dd , J=15, 8.5), 2.98 (1H, s, OH), 5.62 (1H, d, J=15), 6.33 (1H, ddd, J=15, 8.5, 6); MS m/z 398 (M ⁺ , 20) Calculation results for C ₁₉ H ₂₄ F ₆ O ₂ : C, 57.28.H, 6.07; F, 28.61. Measured: C, 57.19; H, 6.25; F, 28.71.

Example 3

This example illustrates that [1R-(1α(E),3aβ,7aα)]]-octahydro-7a-methyl-1-[1-[5,5,5-trifluoro-4-trifluoromethyl The preparation method of ]-4-[(trimethylsilyl)oxy]-2-pentynyl]cyclopropyl]-4H-inden-4-one.

[1R-(1α)(E),3aβ,7aα]]-octahydro-7a-methyl-1-[1-[5,5,5-trifluoro-4-hydroxy-4-tri Fluoromethyl]-2-pentynyl]cyclopropyl]-4H-inden-4-one (165mg, 0.41mmol) in dry _CH2Cl2 (5ml) and _1- (trimethylsilyl) Imidazole (0.5ml, 3.4mmol) was reacted for 5 hours and worked up to give the crude title compound (193mg). Flash chromatography (25 g silica gel, 20% ethyl acetate in hexane) afforded the title compound (161 mg, 83%) as an oil: [α] _D +3.4° CHCl ₃ , c=1.0); IR 1706 cm ⁻¹ ; ¹ H NMR δ0.13(1H, m), 0.22(9H, s), 0.32(1H, m), 0.67(1H, m), 0.70(3H, s), 1.10(1H, m), 1.50-2.40 (11H, m), 2.44 (1H, dd, J=11, 6), 2.68 (1H, dd, J=16, 8.5), 5.57 (1H, d, J=16), 6.16 (1H, ddd, J = 16, 8.5, 6); MS m/z 470 (M ⁺ , 33). _{Anal . Calcd} . for _C22H32F6O2Si : C, 56.15; H, 6.85; F, 24.22. Measured: C, 56.42; H, 6.63; F, 24.37.

Example 4

This example illustrates the preparation of 3-deoxy-1,25-dihydroxy-20-methyl-23-(E)-ene-26,27-hexafluoro-21,28-cyclocholecalciferol.

The Horner reagent [R-(Z)]-[2-[3-[[(1,1-dimethylethyl)-dimethyl-silyl]oxy]-2-methylenecycloethylene Hexyl] ethyl] diphenylphosphine oxide (395mg, 0.872mmol) in anhydrous THF (5.0ml) was decomposed with 1.6M n-butyllithium in hexane (0.55ml, 0.88mmol) at -78°C Proton reaction, after 8 min and ketone [1S-[(1α(E),3aβ,7aα)]]-octahydro-7a-methyl-1-[1-[5,5,5-trifluoro-4- (Trifluoromethyl)-4-(trimethylsilyl)oxy-2-pentynyl]cyclopropyl]-4H-inden-4-one (170mg, 0.361mmol) in anhydrous THF (2.0ml ) solution was reacted for 3 hours and processed. Flash chromatography (45 g silica gel, 20% ethyl acetate in hexanes) gave a gum (215 mg). Dissolve in THF (3ml), treat with 1.0M n- _Bu4N ^{+ F-} in THF (2.8ml), and stir for 19 hours. Further flash chromatography (45 g silica gel, 40% ethyl acetate in hexanes) gave a gum which was dissolved in _HCO2Me (5.0 ml), filtered through a 0.45 [mu]m filter and evaporated. Drying under high vacuum (3 hours) afforded the title compound (144 mg) as a colorless foam: [α] _D 4.0° (methanol, c=0.35); UV λ _max 265 (ε=15,837), 211 (ε=14,458) nm; IR 3598, 1651cm ^-1 ; ¹ H NMR δ0.11(1H, m), 0.29(2H, m), 0.60(3H, s), 0.61(1H, m), 1.10(1H, m), 1.21-1.35 (1H,m), 1.50(6H,m), 1.70(2H,m), 1.90(2H,m), 2.00(3H,m), 2.30(2H,m), 2.60(1H,d, J=12 ), 2.85(2H, m), 2.90(1H, s), 4.22(1H, s), 4.99(1H, s), 5.32(1H, s), 5.42(1H, d, J=12), 5.99( 1H, d, J = 11), 6.10 (1H, ddd, J = 12, 7, 6), 6.36 (1H, d, J = 11); MS (FAB) m/z 535 (M ⁺ +H).

Example 5

This example illustrates the preparation of 3-deoxy-1,25-dihydroxy-20-methyl-23-(Z)-ene-26,27-hexafluoro-21,28-cyclocholecalciferol.

The Horner reagent [R-(Z)]-2-[3-[[(1,1-dimethylethyl)-dimethylsilyl]oxy]-2-methylenecyclohexylene] Ethyl]diphenylphosphine oxide (236 mg, 0.5 mmol) in THF (3.0 mL) was treated with 1.6M n-butyllithium in hexane (0.32 mL, 0.512 mmol). After 8 minutes, the ketone [1S-[1α(Z),3aβ,7aα]]-octahydro-7a-methyl-1-[1-[5,5,5-trifluoro-4-(trifluoromethyl yl)-4-[(trimethylsilyl)oxy]-2-pentynyl]cyclopropyl-4H-inden-4-one (117.5 mg, 0.25 mmol) in THF (2.0 ml) over Stir for 2.5 hours. Work-up gave a gum which was purified by flash chromatography (40 g silica gel, 20% ethyl acetate in hexanes) to give a gum (120 mg). Dissolve in THF (2.0 ml), treat with 1M n- _Bu4N ^{+ F-} in THF (2.0 ml), stir and work up at room temperature for 20 hours. Flash chromatography (40g silica gel, 40% ethyl acetate in hexane) gave the title compound (29 mg) as a colorless foam: [α] _D -41° (methanol, c=0.14); IR 3569cm ^-1 ; UV λ _max 214(10,968), 219(12,931), 259(12,818) nm; ¹ H NMR δ 0.02(1H, m), 0.32(2H, m), 0.60(1H, m ), 0.61(3H, s), 1.1-1.7(11H, m), 1.85(2H, m), 2.0(3H, m), 2.2(3H, m), 2.85(3H, m), 4.12(1H, br s), 4.90 (1H, s), 5.30 (1H, s), 5.40 (1H, d, J=12.8), 6.0 (1H, d, J=20 11), 6.12 (1H, dd, J=12.8 , 6.8), 6.29 (1H, d, J=11), 6.12 (1H, dd, J=12.8, 6.8), 6.29 (1H, d, J=11); MS m/z 518 (M ⁺ , 22) .

Example 6

This example illustrates the preparation of 3-deoxy-1,25-dihydroxy-20-methyl-23-yne-21,28-cyclocholecalciferol.

The Horner reagent [R-(Z)]-2-[3-[[(1,1-dimethylethyl)-dimethylsilyl]oxy]-2-methylenecyclo-hexylene A solution of ]ethyl]diphenylphosphine oxide (375mg, 0.828) in THF (5.0ml) was treated with 1.6M n-butyllithium in hexane (0.51ml, 0.81mmol). After 8 minutes, add [1R-[1α, 3aβ, 7aα]]-octahydro-7a-methyl-1-[4-methyl-4-[(trimethylsilyl)oxy]-2-pentane Alkynyl]cyclopropyl]-4H-inden-4-one (180 mg, 0.50 mmol) in THF (4.0 mL) and the mixture was worked up for 3.5 hours. Flash chromatography (45 g silica gel, 7% ethyl acetate in hexane) gave a syrup (273 mg), which was dissolved in THF (3.3 ml) and together with 1.0 M n-Bu ₄ N ⁺ F ^- in THF (3.3 ml) Stir for 28 hours and work up. After flash chromatography (45g silica gel, 40% ethyl acetate in hexane) chromatography, the title compound (114 mg) was obtained as a colorless foam: [α] _D -70.32° (ethanol, c=0.539); UV λ _max 215 (ε=13,326), 262(ε=17,661) nm; IR 3601cm ^-1 ; ¹ H NMR δ0.28(2H, m), 0.41(1H, m), 0.59(1H, m), 0.60(3H, s ), 1.10(1H, m), 1.50(6H, s), 1.55-2.0(18H, m), 2.09(1H, d, J=17), 2.22(2H, m), 2.60(1H, d, J = 17), 2.80 (1H, d, J = 11), 4.11 (1H, br s), 4.91 (1H, s), 5.30 (1H, s), 5.99 (1H, d, J = 11), 6.27 ( 1H, d, J=11); MS m/z 408.3 (M ⁺ , 60).

Example 7

This example illustrates the preparation of 3-deoxy-1,25-dihydroxy-20-methyl-23-yne-26,27-hexafluoro-21,28-cyclocholecalciferol.

The Horner reagent [R-(Z)]-2-[3-[[(1,1-dimethylethyl)-dimethylsilyl]oxy]-2-methylenecyclohexylene] Ethyl]diphenylphosphine oxide (350mg, 0.773mmol) in THF (5.0ml) was deprotonated with 1.6M n-butyllithium hexane solution (0.49ml, 0.784mmol) at -78°C, 8 Minutes later, and ketone [1S-1α, 3aβ, 7aα]]-octahydro-7a-methyl-1-[1-[5,5,5-trifluoro-4-hydroxy-4-(trifluoromethyl )-2-pentynyl]cyclopropyl-4H-inden-4-one (158 mg, 0.40 mmol) was reacted for 3.0 hours. The crude product from flash chromatography (45 g silica gel, 20% ethyl acetate in hexane) was desilylated with 1 M n-Bu ₄ N ⁺ F ^- in THF (1.8 ml) at room temperature for 18 After 1 hour, purification by flash chromatography (45 g silica gel, 40% ethyl acetate in hexane) gave the title compound (117 mg) as a colorless foam: [α] _D -58.3° (ethanol, c=0.456); UV λ _max 214 (ε=12,900), 260 (ε=15,701) nm; ¹ H NMR δ0.29 (1H, m), 0.35 (1H, m), 0.37 (1H, m), 0.59 (3H, s), 0.64 ( 1H, m), 1.4-1.90 (12H, m), 2.00 (4H, m), 2.18 (1H, d, J=17), 2.25 (2H, m), 2.72 (1H, d, J=17), 2.81 (1H, m), 3.34 (1H, s, OH), 4.12 (1H, br s), 4.92 (1H, s), 5.28 (1H, s), 5.98 (1H, d, J=11), 6.27 ( 1H, d, J=11); MS m/z 516.2 (M ⁺ , 90).

Example 8

This example illustrates the preparation of 3-deoxy-1,25-dihydroxy-20-methyl-21,28-cyclocholecalciferol.

Charge 0.564g (1,246mmol) of [R-(Z)]-2-[3-[[(1,1-dimethylethyl)-dimethylformaldehyde into a magnetically stirred 25ml three-necked round-bottomed flask Silyl]hydroxy]-2-methylenecyclohexylidene]diphenylphosphine oxide, the flask was equipped with a thermometer, a rubber septum on the side and a Claisen fitting in the center with a nitrogen purge and rubber septum. The material was dried under high vacuum and 5ml of tetrahydrofuran was added; the solution was stirred and cooled to -70°C and 0.78ml (1.246mmol) of a 1.6M solution of butyllithium in hexane was added. (The red color disappears after the initial addition of 0.16ml). The solution was stirred at -70°C for 10 minutes, then 0.2904 g (0.796 mmol) [1S-(1α, 3aβ, 7aα)] octahydro-7a-methyl-1-[1- [4-Methyl-4-oxyl]-2-pentyl-cyclopropyl]-4H-inden-4-one. When the reaction was finally complete (TLC, 1:9 ethyl acetate-hexane), the mixture was warmed to -30°C, and then 12 ml of pH 7 phosphate buffer (139.4 g of dipotassium hydrogen phosphate in 400 ml aqueous solution plus 10ml 2M phosphoric acid). The mixture was stirred for 5 minutes, then transferred to a separatory funnel with 35 mL of hexane. The aqueous phase was extracted again with 30 ml of hexane. The two hexane layers were combined, washed with 20 mL of brine, dried over a sodium sulfate plug, and evaporated to a colorless syrup. This material was dissolved in hexane. When solids appear, the hexane suspension must be filtered with a flash tower 25 x 120 mm. After the second portion (20ml each) the mobile phase was changed to 1:79 ethyl acetate-hexane. Fractions 11-18 (TLC according to 1:19 ethyl acetate-hexane) were collected and evaporated. This gave 0.4202 g (88.1%) of the silylated title compound.

A 100 ml brown round bottom flask was charged with 0.4202 g of the silylated title compound. To this material was added 5 ml of tetrahydrofuran and 3.5 ml of a 1M solution of tetrabutylammonium fluoride in tetrahydrofuran and stirred at room temperature for 17 hours. TLC (1:1 ethyl acetate-hexanes and ethyl acetate) showed one major spot. The reaction mixture was then diluted with 13 mL of brine, stirred for 15 minutes, and transferred to a separatory funnel with 40 mL of ethyl acetate. The aqueous layer was extracted again with 20 ml of ethyl acetate. The two organic layers were combined, washed with 5 x 35 mL of water and once with brine, then passed through a plug of sodium sulfate and evaporated to give 0.3523 g of crystalline white residue. The material was chromatographed on a 25 x 110 mm column with a mobile phase of 1:1 ethyl acetate-hexane. Fractions 3-4 had crystallized in the tube. The resulting suspension was concentrated to a volume of about 5 ml, diluted with hexane, concentrated and filtered to give 0.2567 g of the crystalline title compound. [α] _D ²⁵ -59.1° (c 0.325, ethanol). UV λ _max (methanol): 214, 262; λ _sh 222nm. _Anal. Calcd. for _C28H44O : C, 81.50; H, 10.75; O, 7.75. Found: C, 81.43; H, 10.69; O, 7.48.

Table 1: Representative compounds of structural formula I

The unshared valences on the oxygens in the table above will be occupied by hydrogens (the unpaired electrons on the oxygens will bond with the hydrogens).

Example 9

This example illustrates the synthesis of 3-deoxy-1α-acetoxy-25-hydroxy-20-cyclopropyl-23E-ene-26,27-hexafluoro-cholecalciferol.

A 25 ml round bottom flask was charged with 0.1702 g of 3-deoxy-1α,25-dihydroxy-20-cyclopropyl-23E-ene-26,27-hexafluoro-cholecalciferol and 2.85 g of pyridine. The solution was stirred and cooled in an ice bath, then 0.5 mL of acetic anhydride was added and stirring was continued for 1 hour in the ice bath. The solution was then left overnight in the refrigerator. The flask was returned to the ice bath and 0.2 mL of acetic anhydride was added. After 6 h the solution was diluted with 10 mL of ethyl acetate and while still immersed in the ice bath, 2 mL of water was added. The mixture was stirred for 5 minutes, then transferred to a separatory funnel with 20 mL of water and 20 mL of 1:4 ethyl acetate-hexane. The aqueous phase was extracted again with 10 mL of 1:4 ethyl acetate-hexane. TLC of 1:2 ethyl acetate-hexanes showed no product in the second extract. Therefore, the crude extract was washed with 4 x 20 mL of water and 10 mL of brine, then passed through a sodium sulfate plug and evaporated. The residue was dissolved in 1:4 ethyl acetate-hexane, and subjected to flash chromatography on a 15×150 mm column with a mobile phase of 1:4 ethyl acetate-hexane, and each 10 mL was regarded as a fraction. The bulk of the main product (0.105 g) was in fraction 3. Fraction 3 also contained traces of the fast eluting material observed in Fraction 2. Attempts were made to dissolve the residue in 1:6 ethyl acetate-hexane and crystallization began. A small amount of pentane was therefore added and the mixture was allowed to crystallize in the refrigerator. The mother liquor was removed and evaporated to yield 0.0068g. The crystals were washed once with pentane and dried to obtain 0.0899 g of the title compound.

[α] _D -30.8° (54%, methanol),

NMR ( _CDCl3 ): 2.07 (OAc),

Analysis and calculation results: C 64.27, H 6.83. Measured: C 64.63, H 6.93; C 64.52, H 6.94

UV max, nm (absorbance) 121 (0.3639), 250 (0.4591), 262 (4486), 244sh (0.4482).

Example 10

This example illustrates the synthesis of 3-deoxy-1α,25-diacetoxy-20-cyclopropyl-23E-ene-26,27-hexafluoro-cholecalciferol.

Charge 0.315 g (0.6 mmol) 3-deoxy-1α,25-dihydroxy-20-cyclopropyl-26,27-hexafluoro-cholecalciferol, 60 mg dimethylaminopyridine, 5 mL pyridine into a 25 mL round bottom flask , the flask was equipped with magnetic stirring and a Claisen connection with a nitrogen purge and stopper. The flask was soaked in an ice bath and pyridine was added. After 10 minutes, 1.0 mL of acetic anhydride was added dropwise, and stirring was continued in an ice bath. The starting material was completely undetectable after 1 hour. After two hours, the solution was diluted with 10 mL of ethyl acetate while still soaking in the ice bath, and after 10 minutes 2 mL of water was added dropwise. The mixture was stirred for 10 minutes, then transferred to a separatory funnel with 20 mL of water and 20 mL of 1:4 ethyl acetate-hexane. The aqueous phase was extracted again with 10 mL of 1:4 ethyl acetate-hexane. TLC (1:2 ethyl acetate-hexanes) showed no product in the second extract. Therefore, the crude extract was washed with 4 x 20 mL of water and 20 mL of brine, then passed through a sodium sulfate plug and evaporated. The residue was dissolved in 1:6 ethyl acetate-hexane, and subjected to flash chromatography on a 25×150 mm column with a mobile phase of 1:6 ethyl acetate-hexane, and each 20 mL was regarded as a fraction. Pure material was contained in fraction 3 and fractions 4-6 contained product spiked with slower eluting material (TLC 1:2). Therefore, fractions 4-6 were combined and subjected to 15×150 mm column chromatography with a mobile phase of 1:9 ethyl acetate-hexane, and each 20 mL was regarded as a fraction. Fractions 2 and 3 contained the bulk of pure product. These fractions were added to fraction 3 from the first chromatography and evaporated. The residue was dissolved 1:9, filtered and concentrated, diluted with pentane and frozen. The mother liquor was recovered, the crystals were washed with pentane, and dried under high vacuum for 2 hours to obtain 0.190 g of the title compound.

NMR (CDCl ₃ ): δ 2.08 and 2.20 (20Ac),

[α] _D -110.3, 0.59% (methanol),

MA 192020: found C, 63.99; H, 6.62; calculated C, 63.78; H, 6.69,

UV max (absorbance) 212 (0.3573), 251 (0.4392), 261 (0.4276).

Example 11

This example illustrates the method for determining the anabolic effect of the compound of the present invention on rat bone.

Three-month-old rats were ovariectomized (Ovx), and the drug was 1,25-dihydroxyvitamin D ₃ or a compound of the present invention, which was administered by mouth every day after three weeks of ovariectomy until six weeks after ovariectomy. Rats were sacrificed one week later. Control groups included shams (rats that were not ovariectomized) and an Ovx group that received vehicle only. Blood and urine samples were collected twice, once at 4 weeks and once at 6 weeks after ovariectomy, to measure serum and urine calcium concentrations. Final femoral calcium concentrations were determined six weeks after ovariectomized sacrifice.

Bone mineral density of the right femur was determined using the High Resolution Software Package on a QDR-1000W Bone Densitometer (Hologic, Walthan, MA). The animal to be scanned was placed on the scanning block in a supine position with its right leg perpendicular to the trunk and the tibia perpendicular to the femur.

Example 12

This example illustrates the assay method for comparing the in vivo effects of the compound of the present invention and 1,25-(OH) ₂ vitamin D ₃ .

The effects of the compounds of the present invention and 1,25-dihydroxyvitamin _D3 can be compared using the ovariectomized rat model, a standard animal model of postmenopausal osteopenia. Three-month-old rats were ovariectomized and Ovx treatment was started one week after treatment for 8 weeks. The drug in a miglyol (medium chain triglyceride) vehicle is administered orally once daily. Blood and urine samples were collected at 3 and 6 week time points, and bone mineral density (BMD) was measured in vivo after 6 weeks using Dual Energy X-ray Absorptiometry (holographic QDR-4500 ^TM bone scanner). The animals were sacrificed at the 8th week, and the lumbar vertebrae and femurs were taken out for in vitro [ex vivo] BMD determination (Lunar PiXiMus ^TM Bone Scanner) and strength biomechanical tests. The data for each compound were analyzed to determine the highest safe dose. The highest safe dose is defined as the dose that does not produce hypercalcemia (serum calcium concentration exceeding 10.0 mg/dl).

Example 13

This example illustrates the results of a comparison of the in vivo effects of Compound 27 and 1,25-(OH) _{2Vitamin D3} .

The standard animal model of postmenopausal osteopenia, rat ovariectomized model, can be used to compare the effect of Compound 27 and 1,25-dihydroxyvitamin D ₃ (ie VD ₃ ). Three-month-old rats weighing 285 g were ovariectomized (OVX), one week after OVX, and treated for 8 weeks. The drug in a miglyol (medium chain triglyceride) vehicle is administered orally once daily. Blood and urine samples were collected at 3- and 6-week time points, and bone mineral density (BMD) was measured in vivo after 6 weeks using Dual Energy X-ray Absorptiometry (holographic QDR-4500 ^TM bone scanner). The animals were sacrificed at the 8th week, and the lumbar vertebrae and femurs were taken out for in vitro BMD determination (Lunar PiXiMus ^TM Bone Scanner) and strength biomechanical tests. The data for each compound are given in the corresponding table to determine the highest safe dose. The highest safe dose is defined as the dose that does not produce hypercalcemia (serum calcium concentration exceeding 10.0 mg/dl).

Table 3 shows the safety (serum calcium, urine calcium) measurements. The data compared all showed that the blood calcium concentration was 9.3 mg/dl at 3 weeks and 9.8±0.1 mg/dl at 6 weeks. At these doses, the urinary calcium excretion samples per 24 hours were identical for the 0.4 nmol/kg dose of VD ₃ and the 0.5 nmol/kg dose of Compound 27. Table 4 lists the efficacy parameters (BMD, biomechanics, and urinary deoxypyridinolines) for both compounds. BMD at all bone locations listed was higher in animals treated with Compound 27 than in animals treated with VD ₃ (both The two drugs produced equivalent blood calcium concentrations) were significantly higher. These doses represent those expected to safely achieve maximal efficacy, whereas larger doses of each compound are known to cause hypercalcemia. Urinary deoxypyridinium, A marker of bone resorption, the concentration of compound 27 at week 6 was significantly lower than that of VD ₃ , indicating that compound 27 has a stronger ability to inhibit bone resorption. Compound 27 treatment (treatment) formed Stronger spine. The breaking load required to fracture the L5 vertebrae was determined using the axial crush test. At the highest safe dose, animals treated with compound 27 required significantly more force to fracture the vertebrae than those treated with VD ₃ treated animals.

Table 3, Compound 27 and the safety comparison of 1,25-dihydroxy-vitamin D ₃ (VitD) parameter Counterfeit OVX control VitD0.012nmol/kgp vs control VitD0.04nmol/kgp vs control Compound 270.5nmol/kgp vs control p vs 0.012VD3 p vs 0.04VD3 blood & urine calcium Serum calcium (mg/ml) 3 weeks 6 weeks Average SEMn/group Average SEMn/group 9.40.1109.50.110 9.20.1109.30.11.0 9.3ns0.1109.8**+0.110 9.3ns0.1109.7**0.19 9.3ns0.1109.9**+0.210 nsns nsns Urinary calcium (mg/mmol, creatinine/day) 3 weeks 6 weeks Average SEMn/group Average SEMn/group 0.310.02100.340.0310 0.220.05100.260.0510 0.31**0.04100.43**0.0410 0.51**+0.05100.71**++0.059 0.51**+0.06100.72**++0.0710 $$ nsns

ND = no data

ns = no significant difference

^* p<005, ^** p<0.01 vs OVX control

+p<005, ++p<0.01 vs OVX control

$p<0.05, $$p<0.01 vs Vit-D

Table 4, compound 27 and 1,25-dihydroxy-vitamin D ₃ (VitD) effect comparison parameter Counterfeit OVX control VitD0.012nmol/kg VitD0.04nmol/kg Compound 2705nmol/kg p vs0.012VD3 p vs0.04VD3 %BMD > OVX control (in vivo) L2-L4 vertebra L5 vertebra whole femur sublunate femur distal femur 6 weeks 6 weeks 6 weeks 6 weeks 6 weeks Average SEMn/group average SEMn/group average SEMn/group average SEMn/group average SEMn/group 10.01.9911.61.297.31.196.51.5911.71.29 0.01.690.01.290.00.890.01.490.01.19 3.3++1.6102.7++2.3103.3+1.7103.5ns1.7102.8++1.810 5.4*1.694.8+1.491.9++1.790.7++1.592.8++2.09 10.3**2.1108.7**2.0106.3**1.1105.7**1.2108.5**1.310 $$$nsns$$ $ns$$$$$ L5 bone density in vitro BMD(g/cm ² )BMC(g) 8 weeks 8 weeks Average SEMn/group Average SEMn/group 0.1130.002100.0380.00110 0.0980.00270.0330.0017 0.103++0.00390.035ns0.0029 NDND 0.112**0.002100.039**0.00110 $$$$ NDND urine D-PYD (nM/mM creatinine) 3 weeks 6 weeks Average SE Mn/group Average SEMn/group 0.380.04100.310.0310 0.900.06100.840.0710 0.81++0.0990.78++0.0910 0.75++0.0790.76++0.098 0.65*+0.09100.57**+0.0610 ns$ ns$ L5 Vertebral Biomechanical Testing Failure load (N) 8 weeks Average SEMn/group 386327 261206 342*269 ND 419**2310 $ ND

ND = no data

ns = no significant difference

^* p<005, ^** p<0.01 vs OVX control

+p<0.05, ++p<0.01 vs OVX control

$p<005, $$p<0.01 vs Vit-D

The compound of the present invention has biological activity comparable to compound 27.

The present invention has been described in some detail by way of illustration and example for purposes of clarity and understanding. Changes and modifications that are obvious to those of ordinary skill in the art can be made within the scope of the appended claims. Accordingly, it should be understood that the foregoing description is for purposes of illustration, and not limitation. Accordingly, the scope of the invention should be read with reference to the following appended claims, along with the full scope of equivalents to which such claims are entitled.

All patents, patent applications and publications cited in this application are hereby incorporated by reference in their entirety for the purpose of having each individual patent, patent application or publication as if individually indicated.

Claims (16)

1. A compound with the following structural formula:

or its salt, in Dashed lines are optional double bonds; L is a linking group selected from the following groups: -CH ₂ -CH ₂ -CH ₂ -, _-CH2 -CH=CH-, -CH ₂ -C≡C-, _-CH2 - _CH2 -C(=O)-, and -CH=CH-CH=CH-; R ² and R ³ are each independently alkyl or haloalkyl; or R ² and R ³ together with the carbon atoms to which they are attached form a cycloalkyl group; and R and ^R are each independently hydrogen, alkyl, acyl or other ^hydroxyl protecting group, The proviso is that at least one of ^R1 and ^R4 is an acyl group. 2. The compound of claim 1, wherein the structural formula of the compound is as follows in R ¹ , R ² , R ³ , R ⁴ and L are those groups defined in claim 1 . 3. The compound of claim 1, wherein the structural formula of the compound is as follows in L is selected from the following groups: -CH ₂ -CH ₂ -CH ₂ -; _-CH2 -CH=CH-; _-CH2 -C≡C-; and -CH=CH-CH=CH-. 4. The compound of claim 1 or 2, wherein L is selected from _-CH2 -CH=CH-; and _-CH2 -C≡C-. 5. The compound of claim 1 or 2, wherein ^R1 is acyl. 6. The compound of claim 1 or 2, wherein ^R4 is acyl. 7. The compound of claim 1 or 2, wherein ^R2 and ^R3 are each independently selected from alkyl and haloalkyl. 8. The compound of claim 1 or 2, wherein ^R2 and ^R3 are trifluoromethyl. 9. The compound of claim 1 or 2 selected from the group consisting of: 3-deoxy-1α-acetoxy-25-hydroxy-20-cyclopropyl-23E-ene-26,27-hexafluoro-cholecalciferol; and 3-Deoxy-1α,25-diacetoxy-20-cyclopropyl-23E-ene-26,27-hexafluoro-cholecalciferol. 10. A process for the preparation of a compound according to any one of claims 1 to 9, which process comprises: (a) a ketone with the following structural formula: and phosphine oxides of the formula: contacting under conditions sufficient to produce said compound of formula I, in Ar ¹ and Ar ² are phenyl; Dashed lines are optional double bonds; L is a linking group selected from the following groups: -CH ₂ -CH ₂ -CH ₂ -, _-CH2 -CH=CH-, -CH ₂ -C≡C-, _-CH2 - _CH2 -C(=O)-, and -CH=CH-CH=CH-; R ² and R ³ are each independently alkyl or haloalkyl; or R ² and R ³ together with the carbon atoms to which they are attached form a cycloalkyl group; and R and R are ^each independently an alkyl, acyl or hydroxy ^protecting group; and (b) when ^neither R ^nor R is an acyl group, acylation of a compound of formula I with an acylating agent under conditions sufficient to produce a compound of formula I wherein at least one of R ^{and R} is acyl. 11. The method of claim 10, wherein ^R1 and ^R4 are hydroxyl protecting groups. 12. The method of claim 10 or 11, wherein said acylation step (b) comprises: (i) contacting the compound obtained in the step (a) with the hydroxy protecting group scavenging compound under conditions sufficient to produce a 1-hydroxyl-3-deoxyvitamin _D3 analogue of the following structural formula to remove the hydroxy protecting group: and (ii) contacting a 1-hydroxy- ₃ -deoxyvitamin D3 analog and an acylating agent under conditions sufficient to produce a 3-deoxyvitamin _D3 analog ester of the formula: wherein R ^1a is acyl and R ^4a is hydrogen or acyl. 13. The method of claim 12, wherein ^R4a is acyl. 14. A pharmaceutical composition comprising a compound according to any one of claims 1-9 and a pharmaceutically acceptable excipient. 15. Use of the compound according to any one of claims 1 to 9 in the preparation of medicaments for the prevention and treatment of bone-related diseases. 16. The use of claim 15, wherein the bone-related disease is selected from the group consisting of hyperparathyroidism, secondary hyperparathyroidism, renal osteodystrophy and osteoporosis.