KR100327922B1 - Calcitriol Derivatives and Their Uses - Google Patents

Abstract

Methods of treating metabolic bone diseases such as osteoporosis, and other disease states treatable by vitamin D compounds, include administering an effective amount of one or more modified vitamin D compounds. Modified compounds contain conventional vitamin D nuclei but have hydrolyzable groups in vivo at one or more of the 1,3 and 25 carbon positions. The time for converting the modified compound into its active form, such as calcitriol, can be controlled by changing or modifying the hydrolyzable group, thus providing controlled release of the compound over time in vivo.

Description

Calcitriol Derivatives and Their Uses {Calcitriol Derivatives and Their Uses}

The present invention relates to biologically active vitamin D compounds, and more specifically to one of the 1, 3 and 25 carbon positions, such as esters of 1α, 25-dihydroxyvitamin D ₃ or esters of 1,25-dihydroxyvitamin D ₃ analogs. 1,25 (OH) ₂ D ₃ (or 1,25 (OH) in the treatment of vitamin D compounds having a hydrolyzable group and various diseases such as osteoporosis, nephrogenic dysplasia, hypothyroidism or proliferative skin disease ) ₂ D ₃ analogues) for their use to modulate the action over time.

The 1α-hydroxylated metabolites of vitamin D (most importantly 1α, 25-dihydroxyvitamin D ₃ and 1α, 25-dihydroxyvitamin D ₂ ) are known as very potent calcium homeostatic modulators in animals and humans have. The discovery of 1α, 25-dihydroxyvitamin D ₃ as an active vitamin led to a thorough investigation of this hormone type of vitamin D analog to find analogs with selective biological activity. As a result, many structural analogs of these metabolites, such as compounds with different side chain structures, different hydroxylation patterns or different stereochemical characteristics, have been prepared and tested. Important examples of such analogs 1α- hydroxy-vitamin D _3, 1α- hydroxy-vitamin D _2, is the side chain of the fluorinated 1α, 25- dihydroxy various derivatives of vitamin D _3, 19- Nord-vitamin D compound and the side chain Are analogues that are homologous. Some of these known compounds show very potent activity in vivo or in vitro, and some of them have been found to differ interestingly in cell differentiation and calcium regulation. Because of these differences in activity, these compounds have an advantageous therapeutic activity profile, so some of these compounds may not be used in the treatment of a variety of diseases, such as nephropathy, vitamin D-resistant rickets, osteoporosis, psoriasis and certain malignant diseases. Proposed.

Various forms of osteoporosis are known, for example postmenopausal, senile and steroidal osteoporosis, one of the characteristics of which is the loss of bone mass. Postmenopausal women lose significant bone mass and ultimately lead to osteopenia, which in turn causes idiopathic left fractures and iliac fractures of the spine. This disease is commonly known as postmenopausal osteoporosis and is a major medical challenge in the United States and most other countries, where women live 60 to 70 years or older. In general, the disease, which is often accompanied by bone pain and decreased body vertebral activity, is diagnosed by one or two vertebral left fractures that reveal a decrease in bone mass. This disease is known to involve reduced calcium absorption, reduced levels of sex hormones, especially estrogen and androgen, and the opposite calcium balance.

Geriatric osteoporosis and steroidal osteoporosis are also characterized by similar symptoms of bone loss, the latter being known to be the result of long-term glucocorticoid (cortico-steroid) treatment for certain diseases.

There are many ways to treat osteoporosis, but no fully satisfactory treatment is yet known. A common treatment is to administer calcium supplements to the patient. However, calcium supplementation alone does not successfully prevent or treat this disease. Another common treatment is the injection of sex hormones, especially estrogen, which has been reported to be effective in preventing the rapid loss of bone mass in postmenopausal women. However, this technique is difficult to implement due to fear of carcinogenic potential. Other therapies for which inconsistent results have been reported include combinations of large amounts of vitamin D, calcium and fluoride. The main problem with this approach is that fluoride causes a structurally unstable bone called reticulum and also causes a number of side effects such as an increase in fracture frequency and gastrointestinal response to the administered fluoride in large amounts. Another proposed method is to block bone resorption by injecting calcitonin or by feeding phosphonates.

US Pat. No. 4,255,596 proposes the use of various metabolites of vitamin D ₃ to increase calcium absorption and retention in the body of mammals with physiological tendency to show evidence of or decrease in bone mass. However, the specifically named in the above patent metabolites, that 1α- hydroxy-vitamin D _3, 1α- hydroxy-vitamin D _2, 1α, 25- dihydroxy vitamin D ₃ (calcitriol), 1α, 25- dihydroxy Vitamin D ₂ and 1,24,25-trihydroxyvitamin D ₃ may cause hypercalcemia, especially when used with conventional calcium supplements, although the activities described and claimed in the above patents are useful. Calcitriol treatment has also been shown to be effective in reducing bone loss in women with postmenopausal osteoporosis by increasing calcium absorption in the small intestine and decreasing bone resorption. This is described by Aloria et al., "Calcitriol In The Treatment Of Postmenopausal Osteoporosis", Amer. Jour. of Med., Vol. 84, March 1988, pp. 401-408. However, Alleria et al. Oppose the oral administration of calcitriol due to the risk of hypercalcemia. Therefore, the use of calcitriol for the treatment of osteoporosis has not been widely accepted.

Another important consideration is that calcitriol is produced slowly and continuously by the kidneys in vivo and therefore can be used throughout the day. When administered by dose or injection, large amounts may initially be used in tissues, but rarely remain after 2-4 hours due to metabolism and secretion. If there is a way to make calcitriol available in vivo more slowly and more consistently, then the peaks and valleys of calcitriol utilization will be avoided, thus providing the compound at an in vivo effective concentration for an extended period of time and also Sudden use of this overdose prevents or substantially reduces the manifestation of symptoms of hypercalcemia often caused.

Summary of the Invention

The present invention provides methods for modulating and regulating in vivo activity of biologically active vitamin D compounds, such as calcitriol or analogs of calcitriol. More specifically, the present invention provides modified vitamin D compounds which exhibit a desirable and very advantageous aspect of biological activity in vivo, ie that the activity is initiated more gradually and the activity duration is further extended. As a result of such beneficial properties, these compounds represent novel therapeutics for the treatment of all diseases in which vitamin D compounds are effective, such as metabolic bone diseases or proliferative skin diseases (eg, psoriasis), and they represent osteoporosis (postmenopausal, senile) Or steroidal), osteomalacia or renal dysplasia, will prove particularly useful in the treatment of diseases requiring bone formation.

Structurally, an important feature of modified vitamin D compounds having these desirable biological properties is that 1α, 25-dihydroxyvitamins in which the hydrolyzable groups are attached to a hydroxy group at the carbon 25 position, and possibly other hydroxy groups present in the molecule. Derivative of D ₃ or derivative of 1α, 25-dihydroxyvitamin D ₃ analogue. Depending on the various structural factors (e.g., type, size, structural complexity) of the attached group, these derivatives are either 1α, 25-dihydroxyvitamin D ₃ or 1α, 25-dihydroxyvitamin D at different rates in vivo. _It is believed to hydrolyze into analogs of ₃ , causing the biologically active vitamin D compounds (ie, 1,25-dihydroxyvitamin D ₃ or analogs thereof) to be 'slow release' in the body.

Expression "sustained release" in vivo activity profiles of such compounds as well as mixtures of the derivatives (e. G., A mixture of 1,25- dihydroxy vitamin D ₃ derivative or a different 1,25- dihydroxy vitamin D ₃ derivatives of different analogues in) It can be further controlled by using a mixture of a vitamin D compound and at least one vitamin D derivative and not a derivative.

It is important to emphasize that the crucial structural feature of the vitamin derivatives described above is that the hydrolyzable group is attached to a hydroxyl group at the carbon 25 position of the molecule. The presence of a hydrolyzable group at this position allows the resulting derivatives to have the desired 'suspended' biological activity profile mentioned above. Other hydroxy functional groups present in the molecule (eg hydroxy functional groups at the carbon 1 or 3 position) may exist as free hydroxy groups, and one or more of them may be derivatized by hydrolyzable groups. It is not known in the art that the introduction of hydrolyzable groups on carbon 25 of vitamin D molecules significantly alters the biological activity of the resulting derivatives. Recognizing the importance of this particular modification and demonstrating its significant and very beneficial biological effects form the basis of the invention.

The 'hydrolyzable group' present in the abovementioned derivatives is preferably an acyl group, i.e., Q ¹ CO-, wherein Q ¹ is hydrogen or straight, cyclic, branched, saturated or unsaturated Hydrocarbon radicals). Thus, for example, a hydrocarbon radical may be a straight or branched chain alkyl group, a straight or branched chain alkenyl group having one or more double bonds, or a substituted or unsubstituted cycloalkyl or cycloalkenyl group, or a substituted or unsubstituted phenyl, benzyl Or an aromatic group such as naphthyl. Particularly preferred acyl groups are alkanoyl or alkenoyl groups, typical examples of which are formyl, acetyl, propanoyl, hexanoyl, isobutyryl, 2-butenoyl, palmitoyl or oleoyl. Another suitable example of a hydrolyzable group is a group of the type hydrocarbyloxycarbonyl, ie Q ² -O-CO-, wherein Q ² represents a hydrocarbon radical of C ₁ to C ₁₈ as defined above. Examples of such hydrocarbon radicals are methyl, ethyl, propyl, and higher straight or branched chain alkyl and alkenyl radicals, and aromatic hydrocarbon radicals such as phenyl or benzoyl.

Among the modified vitamin D compounds having the preferred in vivo bioactivity profiles shown above, a particularly important and preferred class is certain acyl ester derivatives of calcitriol, ie calcitriol derivatives characterized by the formula:

Wherein X ¹ and X ² independently represent hydrogen or an acyl group, and X ³ represents an acyl group as defined above. Two other very important groups of modified vitamin D compounds are the corresponding acyl esters of calcitriol side chain homologs and acyl derivatives of 19-nor-1,25-dihydroxyvitamin D analogs.

Therefore, the present invention provides a series of modified vitamin D compounds useful for the treatment of metabolic bone diseases (e.g., various forms of osteoporosis, osteomalacia, osteopenia, etc.), or differentiating diseases such as psoriasis or malignant diseases. More specifically, a method of treating such diseases comprises administering an effective amount of the corresponding derivatives represented 1α, 25- dihydroxy vitamin D ₃ derivative or a acyl ester 1α, 25- dihydroxy vitamin D ₃ analogs of the above.

The above compounds may be administered alone or in combination with other pharmaceutically acceptable agents. Doses of about 0.5 μg to about 10 μg per day in individual compounds per se or in combination are generally effective. This method has the distinct advantage that it will restore bone mass as these compounds are converted to calcitriol, which has proven to be effective in treating osteoporosis. Moreover, these compounds will cause less hypercalcemia or hypocalcemia than underivatized compounds, even if administered continuously on a daily basis, as long as the appropriate compound is selected and properly administered. It will be appreciated that the compound and dosage levels will then be adjusted in response to the subject's response monitored by methods known to those skilled in the art.

Since the above methods, including administering these compounds at the indicated doses, are effective at restoring or maintaining bone mass, new methods of treatment or prophylaxis for various forms of osteoporosis, such as postmenopausal osteoporosis, senile osteoporosis and steroidal osteoporosis, have been developed. to provide. In addition to those previously named, it is clear that this method will be readily applied to the prevention or treatment of other disease states with symptoms of bone mass loss. In addition, since the conversion rate of these analogs, i.e., the rate of in vivo hydroxylation reaction, can be controlled and controlled by the method described above, it is certain that this method will be easily applied to the prevention or treatment of hypercalcemia and hypocalcemia. .

1 is a graph illustrating the activity of di- and tri-acetates of 1α, 25-dihydroxyvitamin D ₃ on serum calcium, particularly illustrating the milligram percent of calcium in the blood over time.

FIG. 2 is a graph similar to FIG. 1 except for illustrating the activity of 25-mono-acetate of 1α, 25-dihydroxyvitamin D ₃ on serum calcium.

The present invention provides novel modified vitamin D compounds useful for the treatment of metabolic bone diseases such as osteoporosis, and other disease states. These modified vitamin D compounds can be hydrolyzed in vivo to analogs of calcitriol or calcitriol over a period of time following administration, and as a result, their activity profile in vivo by regulating the bioavailability of the active calcitriol or calcitriol analogues. Also changes. The term 'activity profile' refers to the biological response over time of vitamin D compounds, such as calcitriol or calcitriol analogues. Individual modified compounds or mixtures of such compounds can be administered to fine tune the desired time course of the response.

The term 'vitamin D compound' as used herein refers to the C-ring, D-ring and 3β-hydride of vitamin D, which are linked together by a 5,7 diene double bond system of vitamin D with a side chain attached to the D-ring. And compounds having oxycyclohexane A-rings. In other words, the vitamin D compounds included herein are substituted or unsubstituted A-, C- and D-rings linked together by a vitamin D-specific 5,7 diene double bond system with side chains attached to the D-rings. Includes those that have a 'vitamin D nucleus' consisting of: The term 'modified vitamin D compound' as used herein includes all vitamin D compounds which have been modified by derivatization of one or more hydroxy functional groups present in the vitamin D compound by hydrolysable groups. A 'hydrolyzable group' is a hydroxy modifying group that can be hydrolyzed in vivo to regenerate free hydroxy functional groups.

In the context of the present specification, the term hydrolyzable group preferably denotes acyl and hydrocarbyloxycarbonyl groups, ie, Q ¹ CO- and Q ² -O-CO, where Q ¹ and Q ² have the meanings defined above. Group).

Structurally, the modified vitamin D compounds included in the present invention may be represented by the following formula.

Wherein R ⁵ and R ⁶ each represent hydrogen or collectively represent a methylene group.

The side chain group R in the above formula represents the steroid side chain of the formula

Wherein the stereochemical center (corresponding to C-20 in steroid numbering) can have an R or S configuration (ie, a natural configuration for carbon 20 or vice versa), and Z is Y, -OY,- CH ₂ OY, —C≡CY and —CH═CHY, wherein the double bond may have a cis or trans structure and Y is selected from radicals of the formula

Wherein m and n independently represent an integer from 0 to 5, R ¹ is hydrogen, OX ⁴ , fluoro, trifluoromethyl, and C ₁ -which may be straight or branched and may have a hydroxy substituent _Is selected from ₅ -alkyl, R ² is selected from hydrogen, fluoro, trifluoromethyl, and C _1-5 -alkyl which may be straight or branched and may have a hydroxy substituent, and R ³ and R ⁴ are Independently trifluoromethyl, or C _1-5 alkyl which may be straight or branched and may have a hydroxy substituent, wherein R ¹ and R ² are taken together as an oxo group, or an alkylidene group, = CR ² R ² , Or = CR ² R ³ , or a group-(CH ₂ ) _p- (where p is an integer from 2 to 5), wherein R ³ and R ⁴ join together to a group-(CH ₂ ) _q- (where q is An integer of 2 to 5). As defined above in the present specification, in the formulas shown above, X ¹ , X ² and X ⁴ independently represent a hydrogen, acyl group or hydrocarbyloxycarbonyl group, and X ³ represents an acyl group or hydrocarbyloxycarbonyl group .

Some specific examples of such modified vitamin D compounds are

1α, 25 (OH) ₂ -D ₃ -1,3,25-triacetate, wherein X ¹ = X ² = X ³ = CH ₃ CO,

1α, 25 (OH) ₂ -D ₃ -1,3,25-trihexanoate, wherein X ¹ = X ² = X ³ = CH ₃ (CH ₂ ) ₄ CO,

1α, 25 (OH) ₂ -D ₃ -1,3,25-trinonanoate, wherein X ¹ = X ² = X ³ = CH ₃ (CH ₂ ) ₇ CO, and

Calcitriol derivatives such as 1α, 25 (OH) ₂ -D ₃ -25-acetate, wherein X ¹ = X ² = H and X ³ = CH ₃ CO.

By way of example only, the examples below describe methods of synthesizing modified vitamin D compounds. In these examples, specific compounds distinguished by Arabic numerals (eg, compounds 1, 2, 3..., Etc.) refer to the formulas with the corresponding numbers below. The examples also illustrate the unique biological properties of the novel compounds, such as the underlying properties of using these compounds in the treatment of metabolic bone diseases and other vitamin D responsive diseases such as psoriasis.

1 : X ¹ = X ² = X ³ = H

2 : X ¹ = X ² = X ³ = CH ₃ CO

3 : X ¹ = X ² = X ³ = CH ₃ (CH ₂ ) ₄ CO

4 : X ¹ = X ² = X ³ = CH ₃ (CH ₂ ) ₇ CO

5 : X ¹ = X ² = (CH ₃ ) ₂ CHCO, X ³ = H

6 : X ¹ = X ² = CH ₃ (CH ₂ ) ₄ CO, X ³ = H

7 : X ¹ = X ² = C ₆ H ₅ CO, X ³ = H

8 : X ¹ = X ² = H, X ³ = CH ₃ CO

<Example 1>

Preparation of mono, di and tri-esters of 1α, 25- (OH) ₂ -D ₃

Experiment

Spectra were obtained from the following device.

Ultraviolet (UV) absorption spectra were obtained using a Perkin-Elmer lambda 3B UV / Visible Spectrophotometer. Nuclear magnetic resonance (NMR) spectra were recorded at 400 or 500 MHz using a Bruker DMX spectrometer. Chemical shifts (δ) recorded the downward from internal Me ₄ Si (δ 0.00). Mass spectra were recorded at 70 eV on a Kratos MS-50 TC instrument equipped with a Kratos DS-55 data system. Preparative thin layer chromatography (plc) was performed with a Merck 1-mm F-254 silica gel plate and an E. Merck 0.25 mm F-254 silica gel plate.

General method for the preparation of triester: 1α, 25 (OH) ₂ -D ₃ -1,3,25-triacetate (2) X ^One = X ² = X ³ = CH ₃ Preparation of CO.

A solution of glacial acetic acid (0.05 ml, 0.87 mmol) and trifluoroacetic anhydride (0.05 ml, 0.35 mmol) was stirred at rt for 3.5 h. 1α, 25 (OH) ₂ -D ₃ (1) (0.3 mg, 0.00072 mmol) in 0.4 ml of dry THF was then added to the gastric solution at 0 ° C. The reaction mixture was stirred for 0.5 h at 0 ° C. and then for 2 h at room temperature. The solvent was removed under reduced pressure to give crude product which was purified twice by preparative thin layer chromatography (plc, 15% EtOAc / hexanes) to give pure compound (2) (0.122 mg, 31.3% yield). UV, λ max 265 nm (EtOH). ¹ H NMR (CDCl ₃ ): δ, 0.51 (s, 3H, C ₁₈ -CH ₃ ), 0.92 (d, J = 5.0 Hz, 3H, C ₂₁ -CH ₃ ), 1.42 (s, 6H, C _{26, 27} -CH ₃ ), 1.98, 2.04, 2.06 (s, 9H, CH ₃ CO-), 5.04 (s, 1H, C ₁₉ EH), 5.18 (m, 1H, C ₃ -H), 5.31 (s, 1H , C ₁₉ ZH), 5.48 (t, J = 6 Hz, C ₁ -H), 5.91, 6.35 (dd, J = 10 Hz, C _6,7 -H), MS: m / z 542 (M ⁺ ) 482 (M ⁺ -60), 422 (M ⁺ -2 x 60), 362 (M ⁺ 3 x 60).

1α, 25 (OH) ₂ -D ₃ 1,3,25-trihexanoate (3) X ^One = X ² = X ³ = CH ₃ (CH ₂ ) ₄ CO

To hexanoic acid (0.15 ml, 1.20 mmol) was added trifluoroacetic anhydride (0.1 ml, 0.71 mmol). The mixture was stirred at rt for 4 h (this solution turned pale brown). After addition of 1α, 25 (OH) ₂ -D ₃ ( 1 ) (0.4 mg, 0.00096 mmol) in 0.2 ml of dry THF, the reaction mixture is kept at room temperature for 5 hours, or (monitored by thin layer chromatography) Stir until the reaction is complete. Product 3 (0.217 mg, 31.72% yield) was finished as in the previous example and purified by plc (3 times, 8% EtOAc / Skellysolv B). UV, lambda max. 265.3 nm, 245 nm (shoulder) (EtOH). ¹ H NMR (CDCl ₃ ): δ, 0.51 (s, 3H, C ₁₈ -CH ₃ ), 0.89 (t, J = 6.3 Hz, 9H, CH ₃ (CH ₂ ) ₄ CO), 0.92 (d, J = 6.3 Hz, 3H, C ₂₁ -CH ₃ ), 1.42 (s, 6H, C _26,27 -CH ₃ ), 5.14 (s, 1H, C ₁₉ EH), 5.18 (m, 1H, C ₃ -H), 5.32 (s, 1H, C ₁₉ ZH), 5.47 (t, J = 6.3 Hz, 1H, C ₁ -H), 5.91, 6.34 (dd, J = 11.15 Hz, C _6,7 -H), MS: m / z 710 (M ⁺ ), 594 (M ⁺ -116 C ₅ H ₁₁ COOH), 478 (M ⁺ -2 x 116), 362 (M ⁺ 3 x 116).

1α, 25 (OH) ₂ -D ₃ 1,3,25-trinonanoate (4) X ^One = X ² = X ³ = CH ₃ (CH ₂ ) ₇ CO

Trifluoroacetic anhydride (0.08 ml, 0.566 mmol) was added to nonanoic acid (0.2 ml, 1.145 mmol) and the mixture was stirred at rt for 4 h. After addition of 1α, 25 (OH) ₂ -D ₃ ( 1 ) (0.4 mg, 0.00096 mmol) in 0.2 ml of dry THF, the reaction mixture was completed at room temperature for 4 hours, or by observation (TLC). Stir until. General finishing operation afforded the product ( 4 ) (0.283 mg, 35.2% yield). UV, λ max. 265.3 nm, 243.9 nm (shoulder) (EtOH). ¹ H NMR (CDCl ₃ ): δ, 0.51 (s, 3H, C ₁₈ -CH ₃ ), 0.88 (t, J = 6.2 Hz, 9H, CH ₃ (CH ₂ ) ₇ CO-), 0.92 (d, J = 6.3 Hz, 3H, C ₂₁ -CH ₃ ), 1.42 (s, 6H, C _26,27 -CH ₃ ), 5.04 (s, 1H, C ₁₉ EH), 5.18 (m, 1H, C ₃ -H) , 5.32 (s, 1H, C ₁₉ ZH), 5.49 (t, J = 6.2 Hz, 1H, C ₁ -H), 5.91, 6.34 (dd, J = 11.36 Hz, 2H, C _6,7 -H). MS: m / z 836 (M ⁺ ), 678 (M ⁺ -158, CH ₃ (CH ₂ ) ₇ COOH), 520 (M ⁺ -2 x 158), 362 (M ⁺ 3 x 158).

General method for preparing diesters: 1α, 25 (OH) ₂ -D ₃ 1,3-diisobutyrate (5) X ^One = X ² = (CH ₃ ) ₂ CHCO; X ³ Manufacture of H

Isobutyric acid (0.06 ml, 0.647 mmol), N, N-dicyclohexylcarbodimide (35.55 mg, 0.173 mmol), 4-pyrrolidinopyridine (6.12 mg, 0.041 mmol) and 1α in 0.15 ml of dry dichloromethane A solution of, 25 (OH) ₂ -D ₃ ( 1 ) (0.4 mg, 0.00096 mmol) was stirred at rt overnight (12 h). The white precipitate formed was filtered off and the residue was washed with dichloromethane. The combined organic solutions were concentrated and the crude product was purified twice with plc (25% EtOAc / Skeletal B) to give pure product 5 (296 mg, 55.33% yield). UV, lambda max. 266.5 nm, 244 nm (shoulder). ¹ H NMR (CDCl ₃ ): δ, 0.49 (s, 3H, C ₁₈ -CH ₃ ), 0.93 (d, J = 6.25 Hz, 3H, C ₂₁ -CH ₃ ), 1.14 (t, J = 6.3 Hz, 12H, (CH ₃ ) ₂ CHCO-), 1.21 (s, 6H, C _26,27 -CH ₃ ), 5.05 (s, C ₁₉ EH), 5.17 (m, 1H, C ₃ -H), 5.35 (s , 1H, C ₁₉ ZH), 5.5 (t, J = 6.3 Hz, 1H, C ₁ -H), 5.91, 6.35 (dd, J = 11.35 Hz, 2H, C _6,7 -H). MS: m / z 556 (M ⁺ ), 468 (M ⁺ -88, (CH ₃ ) ₂ CHCOOH), 380 (M ⁺ -2 x 88), 362 (M ⁺ 2 x 88-H ₂ O).

1α, 25 (OH) ₂ -D ₃ 1,3-dihexanoate (6), X ^One = X ² = CH ₃ (CH ₂ ) ₄ CO, X ³ = H

N-hexanoic acid (0.15 ml, 1.19 mmol), N, N-dicyclohexylcarbodimide (73.66 mg, 0.358 mmol), 4-pyrrolidinopyridine (4.45 mg, 0.03 mmol) in 0.1 ml of dry dichloromethane And a solution of 1α, 25 (OH) ₂ -D ₃ ( 1 ) (0.5 mg, 0.0012 mmol) was stirred overnight at room temperature (12 hours). After the finishing operation as in the previous example, the crude product was purified by plc (3 times, 15% EtOAc / skelesolve B) to give pure compound 6 (0.079 mg, 10.74% yield). It was. UV, lambda max. 263.6 nm, 240 nm (EtOH). ¹ H NMR (CDCl ₃ ): δ, 0.51 (s, 3H, C ₁₈ -CH ₃ ), 0.89 (t, J = 6.4 Hz, 6H, CH ₃ (CH ₂ ) ₄ CO-), 0.93 (d, J = 6.25 Hz, 3H, C ₂₁ -CH ₃ ), 1.21 (s, 6H, C _26,27 -CH ₃ ), 5.04 (s, 1H, C ₁₉ EH), 5.18 (m, 1H, C ₃ -H) , 5.30 (s, 1H, C ₁₉ ZH), 5.49 (t, J = 6.2 Hz, 1H, C ₁ -H), 5.92, 6.34 (dd, J = 11.26, 2H, C _6,7 -H), MS m / z 612 (M ⁺ ), 496 (M ⁺ -116, CH ₃ (CH ₂ ) ₄ COOH), 478 (M ⁺ -116-H ₂ O), 380 (M ⁺ 2 x 116), 362 ( M ⁺ 2 x 116-H ₂ O).

1α, 25 (OH) ₂ -D ₃ 1,3-dibenzoate (7), X ^One = X ² = C ₆ H ₅ CO, X ³ = H

Benzoic acid (15.53 mg, 0.127 mmol), N, N-dicyclohexylcarbodimid (42.64 mg, 0.206 mmol), 4-pyrrolidinopyridine (2.13 mg, 0.0144 mmol) and 1α in dry dichloromethane (0.4 ml) A solution of, 25 (OH) ₂ -D ₃ ( 1 ) (0.51 mg, 0.00123 mmol) was stirred at rt for 16 h. After undergoing the finishing operation as in the previous example, the crude product was purified by plc (twice, 8% EtOAc / skelesolve B) to give pure compound 7 (0.123 mg, 16.4% yield). It was. UV, lambda max. 265.4 nm, 230.5 nm (EtOH). ¹ H NMR (CDCl ₃ ): δ, 0.28 (s, 3H, C ₁₈ -CH ₃ ), 0.91 (d, J = 5.0 Hz, C ₂₁ -CH ₃ ), 1.21 (s, 6H, C _26,27- CH ₃ ), 5.13 (s, 1H, C ₁₉ EH), 5.47 (s, 1H, C ₁₉ ZH), 5.51 (m, 1H, C ₃ -H), 5.82 (t, J = 6.2 Hz, 1H, C _1- H), 5.93, 6.45 (dd, J = 10, C _6,7- H), 7.41, 7.54, 8.04 (m, 10H, Ar-H). MS: m / z 624 (M ⁺ ), 606 (M ⁺ -H ₂ O), 502 (M ⁺ -122 C ₆ H ₅ COOH), 380 (M ⁺ -2 x 122), 362 (M ⁺ -2 x 122-H ₂ O).

1α, 25 (OH) ₂ -D ₃ 25-acetate (8), X ^One = X ² = H, X ³ = CH ₃ CO

1α, 25 (OH) ₂ -D ₃ 1,3,25-triacetate ( 2 ) (0.168 mg, 0.00031 mmol) was dissolved in 1 ml of a 0.6% methanol solution of potassium carbonate. The mixture was stirred at room temperature for 2 hours (or until the reaction was completed by TLC measurement) and then the solvent was removed under reduced pressure. The crude product was purified by plc (50% EtOAc / hexanes) to give the pure product ( 8 ) (0.056 mg, 39.4% yield). UV, λ max 265 nm (EtOH). ¹ H NMR (CDCl ₃ ): δ, 0.54 (s, 3H, C ₁₈ -CH ₃ ), 0.92 (d, J = 5 Hz, C ₂₁ -CH ₃ ), 1.42 (s, 6H, C _26,27- CH ₃ ), 1.98 (s, 3H, CH ₃ CO), 4.24 (m, 1H, C ₃ -H), 4.44 (t, J = 6 Hz, C ₁ -H), 5.01 (s, 1H, C ₁₉ EH), 5.34 (s, 1 H, C ₁₉ ZH), 6.02, 6.39 (dd, J = 10 Hz, 2H, C _6,7 -H). MS: m / z 458 (M ⁺ ), 440 (M ⁺ -H ₂ O), 422 (M ⁺ -2 x H ₂ O), 398 (M ⁺ -60, CH ₃ COOH), 380 (M ⁺ - 60-H ₂ O), 362 (M ⁺ -60-2 x H ₂ O).

<Example 2>

This example shows three compounds: 1α, 25 (OH) ₂ D ₃ -1,3,25-triacetate, 1α, 25 (OH) ₂ D ₃ -1,3-diacetate and 1α, 25 The serum calcium response of mice over time for (OH) ₂ D ₃ (not esterified) is shown.

In this biological test, mice were depleted of vitamin D for eight weeks by providing a calcium-containing (0.47% calcium), vitamin D deficiency diet. Rats were then fed each compound at a single oral dose of 1,000 pmol or 1 nanomolar, and rat blood was collected at various times as shown in Table 1 to measure serum calcium. 1 is a graph showing the data of Table 1. FIG. Since calcium is also present in the small intestine of rats as it is present in the diet, an increase in serum calcium mainly indicates calcium absorption in the small intestine. The results clearly show that 1,3-diacetate and unesterified calcitriol exhibit essentially the same hourly response, with the acetylation of C-1 and C-5 hydroxy groups, perhaps the acyl groups at these positions for example It is shown that because it is rapidly removed by the enzyme, it does not significantly change the biological response. In sharp contrast, triacetate did not respond until 12-18 hours after administration and peaked at 24 hours. Thus, the 25-acetate group will remain intact for a prolonged period of time and then slowly hydrolyze inside the body. As above, triacetate clearly delays the use of calcitriol, indicating that the in vivo activity profile of the parent calcitriol can be changed significantly by acylation of the C-25 hydroxy group.

time Control (A) 1.25 (OH) ₂ D ₃ (B) diacetate (C) triacetate 0 hours 4.3 ± 0.07 8 hours 6.06 ± 0.89 5.42 ± 0.90 4.06 ± 0.38 12 hours 7.39 ± 0.10 6.50 ± 0.13 4.70 ± 0.12 18 hours 6.97 ± 0.51 6.24 ± 0.51 5.91 ± 0.30 24 hours 6.45 ± 0.50 5.98 ± 0.43 6.7 ± 0.54 48 hours 4.43 ± 0.18 5.59 ± 0.80 5.11 ± 0.48 5.10 ± 0.43 All data = mean ± standard deviation (B) → (A); No significant difference (C) → (A); P <0.001 at 8 hours and 12 hours

<Example 3>

This example shows two compounds administered in three different methods, oral, intramuscular (IM) and subcutaneous (Sub. Cu.), I.e., 25 (OH) ₂ D ₃ (not esterified). ) And rat serum calcium response over time for 1α, 25 (OH) ₂ D ₃ -25-acetate.

In this test, 1 nanomolar of each compound was given to vitamin D deficient mice fed 0.47% calcium, 0.3% P diet in 0.1 ml of propylene glycol 95% / 5% ethanol. One group consisted of four or more mice. Serum calcium was measured at various times shown in Table 2 according to the single dose administered by the indicated route. 2 is a graph showing the data in Table 2. FIG. The results were in agreement with Example 2. Regardless of the route of administration, the in vivo activity of 1,25 (OH) ₂ D ₃ -25-acetate started more gradually and the activity peak was delayed. Thus, it is evident that 25-monoacetate delays the use of calcitriol, with the presence of C-25-O-acyl groups having a pronounced effect on the activity pattern and response over time of the biologically active vitamin D compound. To prove.

Serum calcium response to 1,25 (OH) ₂ D ₃ and its 25-acetate Serum calcium (mg / 100 ml) compound Route of administration 1 day 3 days 6 days 10 days None (control) 5.9 ± 0.34 - 5.7 ± 0.34 5.2 ± 0.63 1,25- (OH) ₂ D ₃ oral- 9.33 ± 0.41 - 8.6 ± 0.52 7.15 ± 0.68 1,25 (OH) ₂ D ₃ -25-acetate oral- 7.70 ± 0.11 8.87 ± 0.6 8.54 ± 0.15 8.83 ± 0.71 1,25 (OH) ₂ D ₃ -25-acetate Intramuscular 8.10 ± 0.24 9.17 ± 0.53 9.77 ± 0.3 - 1,25 (OH) ₂ D ₃ -25-acetate Subcutaneous 7.78 ± 0.28 9.00 ± 0.28 9.84 ± 0.39 -

Modified vitamin D compounds or combinations thereof can be easily administered in the form of parenteral solutions, either by injection or intravenous, or in oral dosage form, via the digestive tract, or transdermally, or as suppositories. About 0.5 μg per day of the modified vitamin D compound itself, or a combination thereof in combination with other modified vitamin D compounds (the ratio of each compound in the formulation depends on the condition of the particular disease prescribed and the desired degree of bone mineralization and / or bone mobilization) A dosage of from about 10 μg is generally effective for practicing the present invention. In all cases, a sufficient amount of compound should be used for bone mass recovery. In general, amounts of more than about 10 μg of modified vitamin D compounds or other combinations of other modified vitamin D compounds, which are unnecessary to achieve the desired result, can result in hypercalcemia and can be economically unfavorable. In practice, lower doses are generally used for prophylactic purposes, while higher doses are used where the treatment of the disease state is the ultimate goal, and in all given cases the specific dosage administered is known to those skilled in the art. As will be adjusted by the particular compound administered, the disease to be treated, the condition of the subject, and other relevant medical factors that may alter the activity of the drug or the subject's response. In general, both single daily or divided daily administrations are possible, as is known in the art.

Formulations of the various compounds can be prepared by combining non-toxic pharmaceutically acceptable carriers with the compounds into immediate or sustained release formulations, as known in the art. Such carriers can be, for example, both solid or liquid, such as corn starch, lactose, sucrose, peanut oil, olive oil, sesame oil and propylene glycol. When using a solid carrier, the dosage form of the compounds of the present invention may be tablets, capsules, powders, troches and lozenges. When using a liquid carrier, soft gelatin capsules, or syrups or liquid suspensions, emulsions or solutions may be in dosage forms. The dosage form may also contain adjuvant such as preservatives, stabilizers, wetting or emulsifying agents, solution promoters and the like. The formulation may also contain other therapeutically valuable substances.

Modified vitamin D compounds may also include all compounds of the formula below modified by derivatization with one or more hydroxy functional groups that may be present which may be hydrolyzed in vivo.

Where

-P represents hydrogen, alkyl or acyl,

-X represents a portion of the side chain of vitamin D or one of its identified analogs,

-Y and Y 'may be the same or different and represent hydrogen or alkyl, or combine to represent an alkylidene group or form a carbocyclic ring,

-W and W 'may be the same or different and represent hydrogen or alkyl, together represent an alkylidene group or form a carbocyclic ring,

One of the carbon atoms at the center corresponding to positions 14, 13, 17 or 20 may be replaced with oxygen (O), sulfur (S), or nitrogen (NR) with R substituents together with the R and R ′ substituents linked thereto; Can,

-R and R '(ie, R, R ₁ , R ₂ , R' ₂ , R ₃ , R ' ₃ , R ₄ , R' ₄ , R ₅ , R ' ₅ ) are as follows.

R ₁ and R ₃ or R ' ₃ , R ₂ or R' ₂ and R ₄ or R ' ₄ , R ₃ or R' ₃ and R ₅ or R ' ₅ , relative to the center chain relative to 1,3- When positioned, together with three adjacent atoms in the center chain, corresponding to positions 8, 14, 13 or 14, 13, 17 or 13, 17, 20, respectively, R and R ' Saturated or unsaturated carbocyclic or heterocyclic 3-, 4-, 5-, 6- or 7-membered rings may be formed, including when forming a click unsaturated bond,

Provided that R ₁ and R ' ₃ are (1) saturated and unsubstituted, (2) monosubstituted at C-11 or (3) a 6-membered carbocyclic ring having a double bond between C-9 and C-11 When R ₃ is methyl, ethyl or ethenyl, R ₂ and R ₄ do not form a 5-membered carbocyclic ring, or

R ₁ and R ₂ or R ' ₂ , R ₂ or R' ₂ and R ₃ or R ' ₃ , R ₃ or R' ₃ and R ₄ or R ' ₄ , R ₄ or R' ₄ and R ₅ or R 'along the central chain, such as _five relatively located to (i.e., adjacent to the 1,2-position, if not part of a ring as described above, each of positions 8, 14 or 14, 13 or 13, 17 or 17, Saturated or unsaturated carbocyclic or heterocyclic 3-, including when two adjacent atoms in the central chain corresponding to 20 together with R and R 'substituted in pairs together form a cyclic unsaturated bond, May form a 4-, 5-, 6- or 7-membered ring, or

In relative 1,1-positions (ie in pairs) along the center chain, such as R ₂ and R ' ₂ , or R ₃ and R' ₃ , or R ₄ and R ' ₄ or R ₅ and R' ₅ And, if not part of a ring as described above, R and R 'substituents together with the carbon atom to which they are attached a saturated or unsaturated carbocyclic or heterocyclic 3-, 4-, 5- or 6-membered ring Can form,

Which may be the same or different and when they do not form a ring or bond as described above, represent hydrogen or a lower alkyl group or, in the case of a paired substituent, represent a lower alkylidene group.

In the present specification, the expression "lower alkyl group" refers to a straight or branched, saturated or unsaturated carbon chain having 1 to 7 carbon atoms, and the "lower alkylidene group" refers to main chain atoms 14, 13, 17 and / or A straight or branched, saturated or unsaturated carbon chain having 1 to 7 carbon atoms, linked by a double bond to one of 20.

Some of the side chains of vitamin D or one of its identified analogs in the context of the present invention are particularly present in vitamin D ₂ (C-22 to C-28) or D3 (C-22 to C-27), or vitamin D numbering Denotes a substituted alkyl chain having 2 to 15 carbon atoms partially modified as shown below,

In particular one or more, for example having a hydroxyl substituent at positions 24, 25 and / or 26 and / or

One or more, for example with methyl or ethyl substituents at positions 24, 26 and / or 27 and / or

With halogen substituents at one or more positions, eg perfluorinated at positions 26 and / or 27 or defluorinated at positions 24 and / or

The additional substitution of carbon atoms (s) between positions 24 and 25, in particular C ₂₄ , by the same substitutional aspects mentioned above and / or

With ester derivatives of one or more hydroxyl substituents mentioned above and / or

One or more carbons, eg positions 22, 23 or 24, replaced by oxygen, nitrogen or sulfur atoms and / or

-Ringing between carbon atoms 26 and 27 by one bond (cyclopropane), or via 1 to 4 carbon atoms (the ring may be saturated, unsaturated or aromatic and optionally by the aforementioned substituents) May be substituted at all possible position (s)) and / or

Forming a heterocyclic ring comprising an aromatic, wherein between 26 and 27 carbon atoms are cyclized via 1 to 4 atoms, which may be optionally substituted at all possible positions by the aforementioned substituents and / or )

Unsaturated by one or more double or triple C-C bond (s) (the unsaturated chain may be substituted at all possible positions by the aforementioned substituents) and / or

Epoxide functional groups may be present between carbon atoms 22, 23 or 23, 24 or 24, 25 or 25, 26, these epoxidation chains may be saturated or unsaturated and substituted at all possible positions by the aforementioned substituents Can be and / or

Two or more carbon atoms in the side chain are linked by a single bond or via one to five carbon or oxygen, nitrogen or sulfur atoms, including aromatic rings which may be optionally substituted at any possible position by the aforementioned substituents; Forming a 3-7 membered saturated or saturated or unsaturated carbocyclic or heterocyclic ring, and / or

Substituted at one or more positions by saturated or unsaturated carbocyclic, heterocyclic or aromatic rings, which may be substituted at all possible position (s) by the aforementioned substituents,

-Represents the isomeric form of the substituted chain.

Accordingly, the present invention relates to a series of analogues having a wide variety of structures. The compounds of the present invention are mostly represented by one of the following formulae.

Where

X, Y, Y ', W and W' have the same meaning as above,

Z may be all substituted or substituted with heteroatoms such as oxygen, sulfur and nitrogen, in which case Z represents a bond between two 1,3-related carbon atoms in the center chain, 1, 2, 3 Or a saturated or unsaturated hydrocarbon chain consisting of four atoms,

R ₁ , R ₂ , R ' ₂ , R ₃ , R' ₃ , R ₄ , R ' ₄ , R ₅ , R' ₅ may be the same or different and are hydrogen, or methyl, ethyl or n-propyl The same lower alkyl.

Of these, cyclic derivatives of the formula below are preferred.

Where

n is an integer of 2 or 3,

X is vitamin D side chain moieties, i.e. (4-hydroxy-4-methyl) pentyl, (R)-or (S)-(3-hydroxy-4-methyl) pentyl, (3'-hydroxy- 3'-methyl) butyloxy, (4-hydroxy-4-ethyl) hexyl, (4-hydroxy-4-methyl) -2-pentynyl, (4'-hydroxy-4'-ethyl) hexyloxy city; 4,5-epoxy, 4-methyl-2-pentynyl, 4-hydroxy-4-ethyl-2-hexynyl; (3-methyl-2,3-epoxy) -butyloxy; (3-hydroxy-3-ethyl) -pentyloxy; One of (4-hydroxy-4-ethyl) -hexyloxy,

Y, Y ', W and W' represent hydrogen in the same way, or collectively represent a methylene group (= CH ₂ ),

R ₁ , R ₂ , R ' ₂ , R ₃ , R' ₃ , R ₄ , R ' ₄ , R ₅ and R' ₅ may be the same or different and represent hydrogen or methyl.

Claims (23)

1α, 25-dihydroxyvitamin D ₃ -1,3,25-trihexanoate. 1α, 25-dihydroxyvitamin D ₃ -1,3,25-trinonanoate. 1α, 25- dihydroxy vitamin D ₃ -25- formate. Selecting vitamin D compounds having a hydroxy group at carbon 25 and having the desired therapeutic activity in vivo, Acyl or hydro groups which, when attached to an oxygen atom of a hydroxyl group of carbon 25, hydrolyze in vivo at a rate desirable for the active vitamin D to provide a modified vitamin D derivative that extends the in vivo active profile of the active vitamin D. Selecting a carbyloxycarbonyl group, and Modifying the vitamin D compound by inducing a hydroxyl group of carbon 25 to the acyl or hydrocarbyloxycarbonyl group to obtain the modified vitamin D derivative having the formula How to adjust the activity profile of the vitamin D compound comprising a. [Wherein R ⁵ and R ⁶ each represent hydrogen, or R ⁵ and R ⁶ together represent a methylene group, Z is selected from Y, -OY, -CH ₂ OY, -C≡CY and -CH = CHY Wherein the double bond may have a cis or trans stereochemical configuration, and Y is selected from radicals of the formula Wherein m and n independently represent an integer from 0 to 5, R ¹ may be hydrogen, OX ⁴ , fluoro, trifluoromethyl, and may be straight or branched chain and optionally have a hydroxy substituent Is selected from the group consisting of C _1-5 -alkyl, R ² is hydrogen, fluoro, trifluoromethyl, and C _1-5 -alkyl which may be straight or branched chain and optionally have a hydroxy substituent R ³ and R ⁴ are each selected from trifluoromethyl and C _1-5 -alkyl which may be straight or branched chain and optionally have a hydroxy substituent, and R ¹ and R ² together represent an oxo group or an alkylidene group, = CR ² R ² , = CR ² R ³ , or a-(CH ₂ ) _p -group where p is an integer from 2 to 5, and R ³ and R ⁴ Together represent a-(CH ₂ ) _q -group where q is an integer from 2 to 5, In the above formulas, X ¹ , X ² and X ⁴ are each a hydrogen, acyl group or hydrocarbyloxycarbonyl group, and X ³ is an acyl group or hydrocarbyloxycarbonyl group.] The method of claim 4, wherein X ¹ , X ² , X ³ and X ⁴ independently represent an acyl group or hydrocarbyloxycarbonyl group. The method of claim 4, wherein X ³ is an acyl or hydrocarbyloxycarbonyl group and X ¹ , X ² and X ⁴ each represent hydrogen. The method of claim 4, wherein X ¹ , X ² and X ⁴ are each hydrogen or an acyl group, and X ³ is an acyl group. The method of claim 4 wherein the acyl group is acetyl. The method of claim 4, wherein the acyl group is propanoyl, butanoyl or hexanoyl. The method of claim 4, wherein X ¹ , X ² , X ³ and X ⁴ are each acetyl. The method of claim 4, wherein X ¹ , X ² and X ⁴ are each hydrogen and X ³ is acyl. The method of claim 11, wherein X ³ is acetyl. The method of claim 4, wherein the induced vitamin D compound has the formula: Wherein X ¹ and X ² are each a hydrogen, acyl group or hydrocarbyloxycarbonyl group and X ³ represents an acyl group or hydrocarbyloxycarbonyl group. The method of claim 13, wherein X ¹ and X ² are each hydrogen or an acyl group, and X ³ is an acyl group. The method of claim 14, wherein the acyl group is acetyl. Selecting a vitamin D compound having an —OH group attached to carbon 25 and having a therapeutically active profile in vivo, and (a) selecting a -OX ³ group in which X ³ can be hydrolyzed at a desired rate to the -OH group in vivo, and (b) providing said vitamin D compound in modified form having a -OX ³ group attached to carbon 25. Prolonging the activity profile of the vitamin D compound by A method of adjusting the active profile of vitamin D, comprising. The method of claim 16, wherein the hydrolyzable group X ³ is an acyl group. The method of claim 16, wherein the hydrolyzable group X ³ is CH ₃ CO. The method of claim 16, wherein the hydrolyzable group X ³ is CH ₃ (CH ₂ ) ₄ CO. The method of claim 16, wherein the hydrolyzable group X ³ is CH ₃ (CH ₂ ) ₇ CO. The method according to claim 16, wherein the hydrolyzable group X ³ is a hydrocarbyloxycarbonyl group. The method of claim 16, wherein the hydrolyzable group X ³ is formyl. The method of claim 16, wherein the vitamin D has the formula: [Wherein R ⁵ and R ⁶ each represent hydrogen, or R ⁵ and R ⁶ together represent a methylene group, Z is selected from Y, -OY, -CH ₂ OY, -C≡CY and -CH = CHY Wherein the double bond may have a cis or trans stereochemical configuration, and Y is selected from radicals of the formula Wherein m and n independently represent an integer from 0 to 5, R ¹ may be hydrogen, OX ⁴ , fluoro, trifluoromethyl, and may be straight or branched chain and optionally have a hydroxy substituent Is selected from the group consisting of C _1-5 -alkyl, R ² is hydrogen, fluoro, trifluoromethyl, and C _1-5 -alkyl which may be straight or branched chain and optionally have a hydroxy substituent R ³ and R ⁴ are each selected from trifluoromethyl and C _1-5 -alkyl which may be straight or branched chain and optionally have a hydroxy substituent, and R ¹ and R ² together represent an oxo group or an alkylidene group, = CR ² R ² , = CR ² R ³ , or a-(CH ₂ ) _p -group where p is an integer from 2 to 5, and R ³ and R ⁴ Together represent a-(CH ₂ ) _q -group where q is an integer from 2 to 5, and In the formulas, X ¹ , X ² and X ⁴ are each hydrogen or a hydrolyzable group, and X ³ represents hydrogen.]