JP3372259B2 - Calcitriol derivatives and their uses - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION BACKGROUND OF THE INVENTION The present invention relates to biologically active vitamin D compounds, and more particularly, l [alpha], 25-esters of shea hydroxyvitamin D ₃ or 1,25-dihydroxyvitamin D ₃ Vitamin D compounds with hydrolyzable groups at one or more of 1,3 and 25 carbon positions, such as analog esters, and osteoporosis, renal osteodystrophy, hypoparathyroidism or proliferation 1,25 (O 2
H) ₂ D ₃ (or 1,25 (OH) ₂ D ₃ analogues) relates to their use to regulate the action over time.

Vitamin D 1α-hydroxylated metabolites--most importantly 1α, 25-dihydroxyvitamin D ₃ and 1α, 25-dihydroxyvitamin D ₂₋ are very useful modulators of calcium homeostasis in animals and humans. It is known as a substance. With the discovery of 1α, 25-dihydroxyvitamin D ₃ as the active form of this vitamin, a dedicated study of this hormone form of the vitamin D analog with the aim of discovering analogs with selective biological activity. Was done. As a result, many structural analogs of these metabolites have been produced and tested, such as compounds with different side chain structures, different hydroxylation patterns, or different stereochemistry. Important examples of such analogues are 1α-hydroxyvitamin D ₃ , 1α-hydroxyvitamin D ₂ , various side chain fluorinated derivatives of 1α, 25-dihydroxyvitamin D ₃ , 10-nor-vitamin D compounds, And side chain homologated analogs. It has been found that some of these known compounds show very potent activity in vivo or in vitro, and that some of them show interesting segregation of activity in cell differentiation and calcium regulation. Differences in this activity give these compounds an advantageous therapeutic activity profile, so that some of these compounds are associated with renal dysplasia, vitamin D resistant rickets, osteoporosis, psoriasis. , And has been used or proposed for the treatment of various ailments such as certain malignancies.

Various forms of osteoporosis are known, such as post-menopausal, senile and steroid-induced osteoporosis, one of the features of which is loss of bone mass. Menopausal women
It eventually experiences a significant reduction in bone mass that causes osteopenia, which in turn causes idiopathic crush fractures of the vertebrae and fractures of the long bones. This disease is generally
Known as postmenopausal osteoporosis, it poses a major medical problem both in the United States and most other countries where the lifespan of women reaches the age of at least 60 and 70 years. Generally, the disease, often with bone pain and reduced physical activity, is diagnosed by one or two vertebral crush fractures with evidence of reduced bone mass. It is known that this disease is associated with reduced levels of hormones, especially estrogens and androgens, of reduced ability to absorb calcium, and a negative calcium balance.

Similar symptoms of bone loss are characteristic of senile and steroid-induced osteoporosis, the latter of which was approved for long-term glucocorticoid (cortico-steroid) treatment for certain disease states. The result.

Although the methods of treating osteoporosis have changed considerably,
To date, no completely satisfactory therapy is known. A commonly practiced therapy is to administer calcium supplements to patients. However, calcium supplementation alone has not been successful in preventing or curing the disease. Another common therapy is the injection of sex hormones, especially estrogen,
It has been reported to be effective in preventing the rapid loss of bone mass experienced in postmenopausal women.
However, this technique has been complicated by its potential carcinogenic fear. Other therapies with varying results reported included high doses of vitamin D, calciu and fluoride combinations. The first problem with this method is that fluoride induces structurally unhealthy bone, called cancellous bone, which in addition increases the incidence of fractures and the gastrointestinal tract's response to large amounts of administered fluoride. Is to produce many side effects. Another suggested method is to prevent bone resorption by injecting calcitonin or supplying phosphonate.

U.S. Pat.No. 4,255,596, for increasing the absorption and retention of calcium in the body of mammals showing evidence of loss of bone mass or having the physiological tendency of loss of bone mass, use of various metabolites of vitamin D ₃ is suggested. Metabolites specifically named in the above patent, namely 1α-hydroxyvitamin
D ₃ , 1α-hydroxyvitamin D ₂ , 1α, 25-dihydroxyvitamin D ₃ (calcitriol), 1α, 25-dihydroxyvitamin D ₂ and 1,24,25-trihydroxyvitamin D ₃ are described in the above patents. Although the claimed effects are possible, however, they can also cause hypercalcemia, especially if used with commonly used calcium supplements. Calcitriol therapy has also been found to be effective in reducing bone loss in postmenopausal women with osteoporosis by increasing intestinal calcium absorption and decreasing bone resorption. Aloria
Outside, "Calcitrile in the Treatment-
Of Post Menopausal Osteoporosis (Calcit
riol In The Treatment of Postmenopausal Osteoporos
is) ", Amer. Jour. of Med., Volume 84, March issue, 1988, No. 401-4.
Page 08. However, also outside Aloria, due to the risk of hypercalcemia, water was used for oral administration of calcitriol. Therefore, the use of calcitriol to treat osteoporosis has not been widely accepted.

Another important consideration is that in vivo calcitriol is produced slowly and continuously by the kidneys,
As a result, it can be used all day and night. Large amounts are initially available to tissues when given by mouth or by injection, but scarcely remain after 2-4 hours due to metabolism and excretion. A method by which calcitriol can be made available in vivo more slowly and continuously is to avoid the peaks and valleys of calcitriol availability, thereby allowing this in vivo effective level of It will supply the compound and will also avoid or virtually reduce the episodes of hypercalcemia that often result from the sudden availability of this substance.

SUMMARY OF THE INVENTION The present invention provides methods for modulating and modulating the in vivo activity of biologically active vitamin D compounds, such as calcitriol or analogs of calcitriol. More particularly, the present invention provides modified vitamin D compounds that exhibit a desirable and highly advantageous pattern of biological activity in vivo, ie, a relatively gradual onset and a relatively long lasting activity. As a result of these advantageous properties, these compounds are
It represents a novel therapeutic agent for the treatment of all diseases in which vitamin D compounds such as metabolic bone diseases or proliferative skin diseases (eg psoriasis) have been shown to be effective, and these are osteoporosis. Prove particularly useful for the treatment of diseases in which bone formation is required, such as (menopausal, senile, or steroid-induced), osteomalacia or renal osteodystrophy. Will.

Structurally, the basic feature of modified vitamin D compounds with these desirable biological traits is that they are those in which the hydrolyzable group is a carbon 25 hydroxy group and optionally other hydroxyls present in the molecule. 1α, 25-dihydroxyvitamin D ₃ bound to any of the groups
Or a derivative of 1α, 25-dihydroxyvitamin D ₃ analog. Due to various structural factors of the linking group—eg type, size, structural complexity—these derivatives are at different rates in vivo at 1α, 2.
It is hydrolyzed to 5-dihydroxyvitamin D ₃ or 1α, 25-dihydroxyvitamin D ₃ analogs, which results in biologically active vitamin D compounds in the body (ie 1α, 25
- believed to result in "slow release" dihydroxyvitamin D _3, or analogues thereof).

Vivo activity profile of "sustained-release" of such compounds, of course, mixtures (e.g., 1,25 different derivatives of dihydroxyvitamin D ₃ or 1,25 different derivatives of dihydroxyvitamin D ₃ analogs, derivatives , A mixture of)
Or a mixture of one or more vitamin D derivatives as well as an underivatized vitamin D compound.

It is important to emphasize that the critical structural feature of the vitamin derivative identified above is the presence of a hydrolytic group attached to the carbon 25 hydroxy group of the molecule.
The presence of the hydrolyzing group at the above position provides the desired "extended release" biological activity profile on the resulting derivative. Other hydroxy functional groups present in the molecule (eg hydroxy functional groups of carbon 1 or 3) may be present as free hydroxy groups, or one or more of these may also be derivatized with a hydrolytic group. It may have been done. Carbon 25 of vitamin D molecule
The fact that the introduction of a hydrolyzing group into the protein markedly regulates the in vivo biological activity pattern of the resulting derivative has not previously been recognized. The recognition as a fact of the importance of this particular modification, and the demonstration of its remarkable and highly beneficial biological effect form the basis of the present invention.

The “hydrolyzable group” present in the abovementioned derivatives is preferably an acyl group, ie of the type Q ¹ CO—, where Q ¹ is hydrogen or linear, cyclic, branched, saturated or unsaturated. Represents a hydrocarbon radical of 1 to 18 carbons which can be That is, for example, the hydrocarbon group can be a straight chain or branched chain alkyl group, or a straight chain or branched chain alkenyl group having one or more double bonds, or it can be An optionally substituted cycloalkyl or cycloalkenyl group,
Alternatively, it can be a substituted or unsubstituted phenyl, aromatic group such as benzyl or naphthyl. Particularly preferred acyl groups are alkanoyl or alkenoyl groups, some typical examples of which are formyl, acetyl, propanoyl, hexanoyl, isobutyryl, 2
-Butenoyl, palmitoyl or oleoyl.
Another suitable type of hydrolyzing group is the hydrocarbyloxycarbonyl group, ie of the type Q ² —O—CO—, where Q ² is a C ₁ to C ₁₈ hydrocarbon group as defined above. The base is. Typical examples of such hydrocarbon groups are methyl, ethyl, propyl, and higher linear or branched alkyl and alkenyl groups, and aromatic hydrocarbon groups such as phenyl or benzoyl.

A particularly important and preferred class of modified Vitamin D compounds with the desired bioactivity profile shown above is a particular acyl ester derivative of calcitriol, ie the general structure: A calcitriol derivative characterized in that 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 acyl esters of the corresponding calcitriol side chain homologs, and the acyl derivatives of 19-nor-1,25-dihydroxyvitamin D analogs.

Therefore, the present invention is directed to the treatment of metabolic bone diseases (such as various forms of osteoporosis, osteomalacia, osteodystrophy, etc.) or differentiating diseases such as psoriasis or malignant diseases. Provide a series of modified vitamin D compounds that are useful. More specifically, a method of treating such a disease comprises providing an effective amount of the 1α, 25-dihydroxyvitamin D ₃ acyl ester derivative shown above or the corresponding 1α, 25.
- consists of the administration of derivatives of dihydroxyvitamin D ₃ analogs.

The compounds described above can be administered alone or in combination with other pharmaceutically acceptable agents. Doses of individual compounds per se, or in combination, from about 0.5 μg per day to about 10 μg per day, are generally effective. This method has the distinct advantage that these compounds will restore bone mass as they are converted to calcitriol which has proven to be effective in the treatment of osteoporosis. . In addition, these compounds are advantageously hypercalcemic or less hypodermic than non-derivatized compounds, even if the compounds are continuously administered daily, as long as the appropriate compounds and doses are used. Compounds and dose levels considered unlikely to cause calcemia will be adjusted depending on the patient's response when monitored by methods known to those skilled in the art. That is understood.

The above methods involving administration of the indicated doses of these compounds are effective in restoring or retaining bone mass, resulting in postmenopausal osteoporosis, senile osteoporosis and It provides a novel method of treating or preventing various forms of osteoporosis, such as steroid-induced osteoporosis. It will be apparent that this method could be readily applied to the prevention or treatment of disease states other than the named ones, where loss of bone mass is an indicator. Still further, this method allows the rate of conversion of these analogs, the in-vivo hydroxylation process, to be controlled and regulated as described above, thus preventing hypercalcemia and hypocalcemia. It will also be apparent that it could be readily applied for treatment.

BRIEF DESCRIPTION OF THE FIGURES FIG. 1 is a graph illustrating the activity of 1α, 25-dihydroxyvitamin D ₃ di- and tri-acetate on serum calcium, and in particular, milligrams found in blood over time. FIG. 2 is a graph similar to FIG. 1 except that FIG. 2 shows percent calcium, and FIG. 2 shows the activity of 2-5-mono-acetate of 1α, 25-dihydroxyvitamin D ₃ on serum calcium.

DISCLOSURE OF THE INVENTION The present invention provides novel modified vitamin D compounds that are useful in the treatment of metabolic bone disorders such as osteoporosis as well as other disease states. These modified vitamin D compounds are capable of being hydrolyzed in vivo to calcitriol or an analogue of calcitriol and, as a result, active calcitriol or an analogue of calcitriol in vivo. It regulates the body's availability and thereby also its activity profile in vivo. “Activity profile”
The term refers to the biological response of a vitamin D compound, such as calcitriol or analogs of calcitriol, over time. Individual modifying compounds, or a mixture of such compounds, can be administered to "fine tune" the desired time course of the reaction.

As used herein, the term "vitamin D compound" refers to the C-ring, D-ring and 3β-hydroxy of vitamin D interconnected by the 5,7-diene double bond system of vitamin D. Cyclohexane A-rings as well as compounds having any side chain attached to this D-ring are included. In other words, vitamin D included herein
The compound is a substituted or unsubstituted A-, interconnected by a 5,7-diene double bond system symbolizing vitamin D,
Included are "vitamin D nuclei" consisting of C- and D-rings as well as those having side chains attached to the D-ring. As used herein, the term "modified vitamin D compound" refers to any compound in which one or more of the hydroxy functional groups present in such compounds have been modified by derivatization with a hydrolytic group. Vitamin D compound of. A "hydrolyzable group" is a hydroxy-modifying group that can be hydrolyzed in vivo to regenerate a free hydroxy functional group.

In the context of this disclosure, the term hydrolyzable group is preferably an acyl and hydrocarbyloxycarbonyl group, ie of the type Q ¹ CO— and Q ² —O—CO, respectively, where Q ¹ and Q ² are Groups having the meaning defined above).

Structurally, the included modified vitamin D compounds are
formula: (In the formula, R ⁵ and R ⁶ each represent hydrogen or R ⁵
And R ⁶ together represent a methylene group).

The side chain group R in the structure shown above has the following structure: {Wherein the stereochemical center (C in steroid numbering
-20) corresponds to the R or S configuration [ie carbon 20
Either the natural configuration or the opposite non-natural configuration
It can have, and Z _{is, Y, -OY, -CH 2 OY} , -
C≡CY and —CH═CHY [where the double bond can have a cis or trans configuration, and Y is a structure: Where m and n independently represent an integer from 0 to 5 and R ¹ is hydrogen, OX ⁴ , fluorine, trifluoromethyl, and can be linear or branched. Selected from C _1-5 -alkyl optionally having a hydroxy substituent, and R ² can be hydrogen, fluorine, trifluoromethyl and optionally a straight or branched chain. May have a hydroxy substituent
Selected from C _1-5 alkyl, and R ³ and R ⁴ are independently trifluoromethyl or C, which may be linear or branched and optionally bear hydroxy substituents. _1-5 alkyl, and R ¹ and R ^{2 taken} together are 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), and R ³ and R ⁴ are taken together to form the group — (CH ₂ ) _q — (where q
Represents an integer of 2 to 5) and is selected from the group of]). X ¹ , X ^{2 in} the structure shown above
And X ⁴ separately represent hydrogen, an acyl group or a hydrocarbyloxycarbonyl group, and X ³ represents an acyl group or a hydrocarbyloxycarbonyl group, as previously defined herein.

Some specific examples of such modified vitamin D compounds are: 1α, 25 (OH) ₂ -D ₃ -1,3,25-triacetate [where X ¹ = X ² = X ³ = CH ₃ CO ₃ ] 1α, 25 (OH) ₂ -D ₃ -1,3,25-trihexanoate [where X ¹ = X ² = X ³ = CH ₃ (CH ₂ ) ₄ CO]; 1α, 25 (OH ) ₂ -D ₃ -1,3,25- Turin nano benzoate [wherein ^{X 1 = X 2 = X 3} = CH 3 (CH 2) 7 CO _{]; 1α, 25 (OH) 2} -D 3 -25- Acetate [where X ¹ = X
² = H and X _³ = CH ³ CO]; it is calcitriol derivatives, such as.

The following examples, which are only meant to be illustrative, illustrate the method of synthesis of modified vitamin D compounds. In these examples, certain compounds identified by Arabic numerals (see
(1,2,3, ...) refers to the structure numbered below. Further provided are examples that demonstrate the unique biological characteristics of the novel compounds, such characteristics for the treatment of metabolic bone disease and other vitamin D responsive diseases such as psoriasis. Serve as a basis for the application of these compounds in.

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 1α, 25- (CH) 2} - Production Experiments for D ₃ , Mono, Di and Tri-Esters Spectra were obtained on the following instrument: Ultraviolet (UV) absorption spectra were obtained on a Perkin-Elmer Lambda 3B UV-Visible Spectrophotometer. I took it. Nuclear magnetic resonance (NMR) spectra were recorded at 400 or 500 MHz using a Bruker DMX spectrometer. Chemical shift (δ) reports the shift from internal Me ₄ Si (δ0.00) to the low field side. Mass spectra were recorded at 70 eV on a Kratos MS-50TC instrument equipped with a Kratos DS-55 data system. Preparative thin layer chromatography (plc) is based on Merck 1-
mm F-254 silica gel plates and E. Merk 0.25 mm F-254 silica gel plates.

General manufacturing procedure for triesters: 1α, 25 (OH) ₂ -D
₃ Preparation of 1,3,25-triacetate (2), X ¹ = X ² = X ³ = CH ₃ CO Glacial acetic acid (0.05 mL, 0.87 mmol), trifluoroacetic anhydride solution (0.05 mL, 0.35 mmol) Stir at room temperature for 3.5 hours. Then 1α, 25 (OH) ₂ in 0.4 mL dry THF
-D ₃ (1) (0.3 mg, 0.00072 mmol) was added to the above solution at 0 ° C. The reaction mixture was stirred at 0 ° C. for 0.5 hours and then at room temperature for 2 hours. The solvent was removed under reduced pressure to give a crude product, which was purified twice by preparative thin layer chromatography (plc, 15% EtOAc / hexane) to give pure compound (2) [0.122 mg, 31.3% yield. ) Got.

UV, λ max 265 nm (EtOH). ¹ H NMR (CDCl ₃ ): δ 0.51 (s,
3H, C ₁₈ -CH ₃ ), 0.92 (d, J = 5.0Hz, 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 , 5 19 E-H), 5.18 (m, 1H, C 3 -H), 5.3
1 (s, 1H, C ₁₉ Z-H), 5.48 (t, J = 6Hz, C ₁ -H), 5.91
(6.35 (dd, J = 10Hz , C 6, 7 -H) .MS: m / z 542 (M +), 48
2 (M ⁺ -60), 422 (M ⁺ -2x60), 362 (M ⁺ 3x60). 1α, 25 (OH) ₂ -D ₃ 1,3,25-trihexanoate (3) X ¹ = X ² = X ³ = CH ₃ (CH ₂ ) ₄ CO. Hexanoic acid (0.15 mL, 1.20 mmol) To the mixture was added trifluoroacetic anhydride (0.1 mL 0.71 mmol). The mixture was stirred at room temperature for 4 hours (solution turned light brown).
1α, 25 (OH) ₂ -D ₃ (1) (0.4m in 0.2mL dry THF
g, 0.00096 mmol) was added and the reaction mixture was stirred at room temperature for 5 hours or until completion (as monitored by thin layer chromatography). Product (3) (0.
217 mg, 31.72% yield) was treated as in the previous example to give plc [3 times; 8% EtOAc / Skellyso.
lve) B].

UV, λmax 265.3nm, 245nm (shoulder) (EtOH). ¹ H NMR (CDC
l ₃ ): δ 0.51 (s, 3H, C ₁₈ −CH ₃ ), 0.89 (t, J = 6.3Hz, 9
H, CH ₃ (CH ₂ ) ₄ CO), 0.92 (d, J = 6.3Hz, 3H, C ₂₁ −CH ₃ ),
1.42 (s, 6H, C _26,27 -CH ₃ ), 5.14 (s, 1H, C ₁₉ E-H), 5.1
_{8 (m, 1H, C 3} -H), 5.32 (s, 1H, C 19 Z-H), 5.47 (t, J =
_{6.3Hz, 1H, C 1 -H)} , 5.91,6.34 (dd, J = 11.15Hz, C 6, 7 -
H) .MS: m / z 710 (M ⁺ ), 594 (M ⁺ −116 C ₅ H ₁₁ COOH), 478
(M ⁺ −2x116), 362 (M ⁺ −3x116). 1α, 25- (OH) ₂ -D ₃ 1,3,25-trinonanoate (4) X ¹ = X ² = X ³ = CH ₃ (CH ₂ ) ₇ CO. Nonanoic acid (0.2 mL, 1.145 mmol) Trifluoroacetic anhydride (0.08 mL, 0.566 mmol) was added to the mixture and the mixture was stirred at room temperature for 4 h, 1α, 25 (O in dry THF (0.2 mL).
H) _2- D ₃ (1) (0.4 mg, 0.00096 mmol) was added and then the reaction mixture was stirred at room temperature for 4 hours or until complete (as determined by TLC). The usual work up gave the product (4) (0.283 mg, 35.2% yield).

UV, λmax 265.3nm, 243.9nm (shoulder) (EtOH). ¹ H NMR (CDC
l ₃ ): δ 0.51 (s, 3H, C ₁₈ −CH ₃ ), 0.88 (t, J = 6.2Hz, 9
H, CH ₃ (CH ₂ ) ₇ CO−, 0.92 (d, J = 6.3Hz, 3H, C ₂₁ −C
H ₃ ), 1.42 (s, 6H, C _26,27 −CH ₃ ), 5.04 (s, 1H, C ₁₉ E−
H), 5.18 (m, 1H , C 3 -H), 5.32 (s, 1H, C 19 Z-H), 5.4
9 (t, J = 6.2Hz, 1H, C _1- H), 5.91,6.34 (dd, J = 1.36H
z, 2H, C ₆ , ₇ -H) .MS: m / z 836 (M ⁺ ), 678 (M ⁺ -158, CH ₃
(CH ₂ ) ₇ COOH), 520 (M ⁺ −2x158), 362 (M ⁺ −3x158). General procedure for producing diesters: 1α, 25- (OH) ₂ -D
₃ 1,3-Diisobutyrate (5) X ¹ = X ² = (CH ₃ ) ₂ CHCO; X
Production of ³ = H;

Isobutyric acid (0.06 mL, 0.66 mL) in 0.15 mL of dry dichloromethane.
47 mmol), N, N-dicyclohexylcarbodiimide (35.55 mg, 0.173 mmol), 4-pyrrolidino-pyridine (6.12 mg, 0.041 mmol) and 1α, 25 (OH) ₂ -D ₃
A solution of (1) (0.4 mg, 0.00096 mmol) was stirred overnight (12 hours) at room temperature. The white precipitate that formed was filtered and the residue washed with dichloromethane. The combined organic solution was concentrated and the crude product was purified twice by plc (25% EtOAc / Skellysolve B) to give pure product (5)
(296 mg, 55.33% yield) was obtained.

UV, λ 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 = 6.25Hz, 3H, C 21 -CH
₃ ), 1.14 (t, J = 6.3Hz, 12H, (CH ₃ ) ₂ CHCO−), 1.21 (s,
6H, C _26,27 -CH ₃ ), 5.05 (s, C ₁₉ E-H), 5.17 (m, 1H, C ₃
-H), 5.35 (s, 1H, C ₁₉ Z-H), 5.5 (t, J = 6.3Hz, 1H, C ₁
-H), 5.91,6.35 (dd, J = 11.35Hz, 2H, C 6, 7 -H) .MS: m
^{/ z 556 (M +),} 468 (M + -88. (CH 3) 2 CHCOOH), 380 (M +
−2x88), 362 (M ⁺ −2x88−H ₂ O). 1α, 25 (OH) ₂ -D ₃ 1,3-dihexanoate (6), X ^{1
_{= X 2 = CH 3 (CH}} 2) 4 CO, X 3 = H. 0.1mL n- hexanoic acid in dry dichloromethane (0.15 m
L, 1.19 mmol), N, N-dichlorohexycarbodiimide (73.66 mg, 0.358 mmol), 4-pyrrolidinopyridine (4.45 mg, 0.03 mmol), and 1α, 25 (OH) ₂ −.
A solution of D ₃ (1) (0.5 mg, 0.0012 mmol) was stirred overnight (12 hours) at room temperature. After treatment as in the previous example, the crude product was purified by plc (3 times, 15% EtOAc / Skellysolve B) to give pure compound (6).
(0.079 mg, 10.74% yield) was obtained.

UV, λmax 263.6nm, 240nm (EtOH). ¹ H NMR (CDCl ₃ ): δ 0.51 (s, 3H, C ₁₈ —CH ₃ ), 0.89 (t,
_{J = 6.4Hz, 6H, CH 3} (CH 2) 4 CO -), 0.93 (d, J = 6.25Hz, 3
H, CH ₂₁ -CH ₃ ), 1.21 (s, 6H, C _26,27 -CH ₃ ), 5.04 (s, 1H,
C ₁₉ E-H), 5.18 (m, 1H, C ₃ -H), 5.30 (s, 1H, C ₁₉ Z-
H), 5.49 (t, J = 6.2Hz, 1H, C ₁ -H), 5.92,6.34 (dd, J
_{= 11.26,2H, C 6, 7 -H} ) .MS: m / z 612 (M +), 496 (M + -11
6, CH ₃ (CH ₂ ) ₄ COOH), 478 (M ⁺ −116−H ₂ O), 380 (M ⁺ −2
^{x116), 362 (M + -2x116} -H 2 O). 1α, 25 (OH) ₂ -D ₃ 1,3-dibenzoate (7) X ¹ = X ²
= C ₆ H ₅ CO, X ³ = H. Benzoic acid (15.53 mg, in dry mL (0.4 mL)).
0.127 mmol), N, N-dicyclohexylcarbodiimide (42.64 mg, 0.206 mmol), 4-pyrrolidinopyridine (2.13 mg, 0.0144 mmol), and 1α, 25 (OH) ₂ D ₃
A solution of (1) (0.51 mg, 0.00123 mmol) was stirred at room temperature for 16 hours. After treating as above, the crude product was
Purified by plc (2 times, 8% EtOAc / Skellysolve B) to give pure compound (7) (0.123 mg, 16.4% yield).

UV, λmax 265.4nm, 230.5nm (EtOH). ¹ H NMR (CDCl ₃ ): δ 0.28 (s, 3H, C ₁₈ —CH ₃ ), 0.91 (d,
J = 5.0Hz, CH ₂₁ −CH ₃ ), 1.21 (s, 6H, C _26,27 −CH ₃ ), 5.13
(S, C ₁₉ E-H), 5.47 (s, 1H, C ₁₉ Z-H), 5.51 (m, 1H, C ₃
-H), 5.82 (t, J = 6.2Hz, 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) .M
S: m / z 624 (M +), 606 (M + -H 2 O), 502 (M + -122 C 6 H 5 CO
^{OH), 380 (M + -2x122} ), 362 (M + -2x122-H 2 O). 1α, 25 (OH) ₂ -D ₃ 25-acetate (8) X ¹ = 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 0.6% potassium carbonate solution in methanol. This mixture at room temperature for 2
After stirring for time (or until completion by TLC), the solvent was removed under reduced pressure. The crude product was purified by plc (50% EtOAc / hexane) to give pure product (8) (0.056m
g, 39.4% yield).

UV, λmax 265nm (EtOH). ¹ H NMR (CDCl ₃ ): δ 0.54 (s, 3H, C ₁₈ —CH ₃ ), 0.92 (d,
J = 5.0Hz, CH ₂₁ −CH ₃ ), 1.42 (s, 6H, CH _26,27 −CH ₃ ), 1.9
_{8 (s, 3H, CH 3} CO), 4.24 (m, 1H, C 3 -H), 4.44 (t, J = 6H
z, C ₁ -H), 5.01 (s, 1H, C ₁₉ E-H), 5.34 (s, 1H, C ₁₉ Z-
H), 6.02,6.39 (dd, J = 10Hz, 2H, C 6, 7 -H), MS: m / z 45
8 (M ⁺ ), 440 (M ⁺ −H ₂ O), 422 (M ⁺ −2xH ₂ O), 398 (M ⁺ −6
0, CH ₃ COOH), 380 (M ⁺ −60−H ₂ O), 362 (M ⁺ −60−2xH
₂ O). Example 2 This example illustrates three compounds, namely 1α, 25 (O
H) ₂ D ₃ -1,3,25- _{triacetate, 1α, 25 (OH) 2} D 3 -1,
Figure 3 illustrates the rat serum calcium response over time to 3-diacetate and 1α, 25 (OH) ₂ D ₃ (non-esterified).

In this biological study, rats were fed a calcium-containing (0.47% calcium), vitamin D-deficient diet for 8 weeks to remove vitamin D from these rats. Serum calcium was then determined by feeding them with 1,000 picomoles or 1 nanomolar of each of the above compounds in a single oral dose and collecting blood from these rats at various times as shown in Table 1. FIG. 1 is a graph illustrating the data in Table 1. Elevated serum calcium is mostly indicative of intestinal calcium absorption, as calcium is present in the therapeutic diet and therefore in the intestine.
The results clearly show that 1,3-diacetate and non-esterified calcitriol produce virtually the same time course of reaction, acetylation of the C-1 and C-5 hydroxy groups is probably due to these It specifically demonstrates that the acyl group at the position is not rapidly altered by, for example, digestive enzymes and does not significantly alter the biological response. In sharp contrast, triacetate 12
The reaction did not begin to show up to 18 hours and reached a peak at 24 hours. As a result, the 25-acetate group probably remains intact for a longer period of time and is then slowly hydrolyzed in the body. Thus, triacetate clearly delays the utilization of calcitriol, indicating that the bioactivity profile of the parent calcitriol can be altered very significantly by acylation of the C-25-hydroxy group.

Example 3 This practice uses three different methods: oral, intramuscular (IM), and subcutaneous (Sub.C).
u.), the two compounds administered, namely 1α, 25 (O
H) ₂ D ₃ (non-esterified) and 1α, 25 (O
H) ₂ D ₃ -25- acetate, specifically showing the serum calcium response of rats over the entire time for.

In this test, 1 nmole of each compound was added to 0.47% of propylene glycol 95% / 5% ethanol 0.1m.
Calcium, 0.3% P diet was given to vitamin D deficient rats. There were at least 4 rats per group. Serum calcium was measured at various times shown in Table 2 after a single dose was given by the indicated route. FIG. 2 is a graph illustrating the data in Table 2. This result is consistent with the result of Example 2. Regardless of the route of _{administration, 1,25 (OH) 2 D 3} -25- acetate, shows a peak of activity in more beginning and delays gradual vivo activity. Thus, 25-monoacetate clearly delays the utilization of calcitriol and the presence of the C-25-O-acyl group is apparent in the activity pattern of biologically active vitamin D compounds and in the time course of the reaction. To have a positive effect.

The modified vitamin D compound or a combination thereof is
As a sterile parenteral solution by injection or intravenously,
Or in the oral dosage form by the digestive tract, or transdermally,
Or it can be easily administered by suppository.
The modified vitamin D compound itself or in combination with other modified vitamin D compounds (the ratio of each compound in this combination depends on the particular disease state of interest and the desired bone mineral enrichment). Dependent on the degree and / or the degree of mobilization of bone), a dose of about 0.5 microgram to about 10 microgram of modified vitamin D compound per day is generally effective in practicing the present invention. In all cases, a sufficient amount of compound to restore bone mass should be used. An amount of modified vitamin D compound, or a combination of this compound with other modified vitamin D compounds, in excess of about 10 micrograms per day is generally not required to achieve the desired results,
It will result in hypercalcemia and will not be economically sound practice. In practice, higher doses are used when treatment of the disease state is the desired goal, while lower doses are generally used for prophylaxis and administration in any given case. The particular dose administered will alter the particular compound being administered, the condition to be treated, the condition of the patient and the activity of the drug or the patient's response, as is well known to those skilled in the art. It is understood that it will be adjusted according to other relevant medical facts that can be made. In general, either single or divided daily doses can be used, as is well known in the art.

Dosage forms for various compounds are well known in the art.
They can be prepared by combining them with non-toxic pharmaceutically acceptable carriers to make either immediate release or sustained release formulations. Such carriers are, for example, corn starch,
It can be either solid or liquid such as lactose, sucrose, peanut oil, olive oil, sesame oil and propylene glycol. If a solid carrier is used, the dosage form of the compound may be tablets, capsules, powders,
It may be a lozenge or lozenge. If a liquid carrier is used, it may be a soft gelatin capsule, or a syrup or liquid suspension, emulsion or solution. The dosage form may also contain adjuvants such as preservatives, stabilisers, wetting or emulsifying agents, solubilizers. They can also contain other therapeutically valuable substances.

This modified vitamin D compound may also be present 1
It may also include any of the compounds described below in which one or more hydroxy functional groups have been modified by derivatization with an in vivo hydrolysable group.

-P represents hydrogen, alkyl or acyl; -X represents one of vitamin D or its established analogues.
And -Y and Y'can be the same or different and represent hydrogen or alkyl, or, when taken together, represent an alkylidene group. Or form a carbocycle; W and W'can be the same or different and represent hydrogen or alkyl, or, when taken together, represent an alkylidene group, or One of the central carbon atoms forming a carbocycle and corresponding to the -14, 13, 17 or 20 position
One can be replaced by oxygen (O), sulfur (S) or nitrogen (NR) with an R substituent, together with the R and R'substituents attached thereto; -R and R '(ie , R, R ₁ , R ₂ , R ₂ ′, R ₃ , R ₃ ′, R ₄ ,
R ₄ ′, R ₅ , R ₅ ′) is: R ₁ and R ₃ or R ₃ ′, R ₂ or R ₂ ′ and R ₄ or
When located relatively in the 1,3-position along the central chain, such as R ₄ ′, R ₃ or R ₃ ′ and R ₅ or R ₅ ′, the position
Saturated, unsaturated carbocyclic or heterocyclic 3-, 4-, together with three adjacent atoms of the central chain corresponding to 8,14,13 or 14,13,17 or 13,17,20 , 5-, 6-, or 7-
Can form a member ring, and is also geminal substituted R and R '.
And R ₁ and R ₃ ′ are combined with each other to form a cyclic unsaturated bond, provided that R ₁ and R ₃ ′ are the following properties (1) unsubstituted and saturated (2) monosubstituted by C-11. Or (3) forming a 6-membered carbocycle having a double bond between C-9 and C-11, R ₂ and R ₄ are 5
- membered does not form a carbon ring (this time, R ₃ is methyl, ethyl or ethenyl).

· R ₁ and R ₂ or R ₂ ', R ₂ or R _2' and R ₃ or R
Relative 1,2 along the central chain such as ₃ ′, R ₃ or R ₃ ′ and R ₄ or R ₄ ′, R ₄ or R ₄ ′ and R ₅ or R ₅ ′
-Positioned (i.e., vicinal), and not part of a ring as described above, of the central chain corresponding to positions 8,14 or 14,13 or 13,17 or 17,20, respectively. Together with two adjacent atoms it is possible to form a saturated or unsaturated carbocyclic or heterocyclic 3-, 4-, 5-, 6- or 7-membered ring, which is then also geminally substituted. Also included is the case where R and R ′ together form a cyclic unsaturated bond.

· R ₂ and R ₂ ', or R ₃ and R _3', or R ₄ and
R and R when located relatively in the 1,1-position along the central chain (ie, geminal), such as R ₄ ′ or R ₅ and R ₅ ′, and not part of a ring as described above. ′
Together with the carbon bearing the substituents, saturated or unsaturated carbocyclic or heterocyclic 3-, 4-, 5- or 6-membered rings can be formed.

· May be the same or different, and when they do not form a ring or bond as described above,
Represents hydrogen or a lower alkyl group or, when combined with geminal substitution, represents a lower alkylidene group.

In the context of the present invention, the expression "lower alkyl group" denotes a straight or branched saturated or unsaturated carbon chain containing 1 to 7 carbon atoms and the "lower alkylidene group" is A straight or branched saturated or unsaturated carbon chain containing 1 to 7 carbon atoms linked to one of the main chain atoms 14, 13, 17 and / or 20 through a double bond Indicates.

In the context of the present invention, some of the side chains of one of the vitamin D or its established analogues are especially vitamin D ₂ (C-22
To partially modified as shown below with the numbering of the C-28) or D ₃ (C-22 to either as present on the C-27) in or vitamin D, especially: - One or more positions, such as positions 24, 25 and / or 26,
A hydroxyl substituent and / or one or more positions, such as the 24,26 and / or 27 positions,
A methyl or ethyl substituent and / or a halogen substituent (s) at one or more positions, for example perfluorinated at the 26 and / or 27 positions or difluorinated at the 24 position, and / or Or-an additional carbon atom (s) between positions 24 and 25, in particular C _24, and / or an ester derivative of one or more hydroxyl substituents above, which has the same substitution pattern as above. / Or-exchange one or more carbon atoms, for example carbon atoms in the 22,23 or 24 position with oxygen, nitrogen or sulfur atoms, and /
Or — cyclized by a bond (cyclopropane) or by intervening 1 to 4 carbon atoms between carbon atoms 26 and 27, the ring being saturated, unsaturated or aromatic And optionally substituted at any possible position with the above-mentioned substituents, and / or-cyclizing between carbon atoms 26 and 27 with 1 to 4 atoms to form an aromatic ring To form a heterocycle, which is optionally substituted at any possible position with the above-mentioned substituents, and / or has one or more double or triple C—C bonds. Are unsaturated and these unsaturated chains can be substituted at any possible position by the above substituents,
And / or-epoxide functional groups may be present between carbon atoms 22,23 or 23,24 or 24,25 or 25,26; these epoxidized chains may be saturated or unsaturated And can be substituted at any possible position by the above substituents, and / or-two or more of the carbon atoms of the side chains are bonded by a single bond, or
To 5 or 5 carbons or oxygen, nitrogen or sulfur atoms may combine to form a 3-7 membered saturated or unsaturated carbocycle or heterocycle (including aromatic rings), which ring is Can be substituted at any possible position by the above-mentioned substituents, and / or-at one or more positions saturated, unsaturated carbocycle, heterocycle or aromatic ring [these being either Substituted at the possible position (s) also with the above-represented substituents-a substituted alkyl chain of 2 to 15 carbon atoms in an isomeric form of the substituted chain Represent.

Thus, the present invention relates to a series of analogs with widely varying structures.

Most often the compound of the invention has the formula [Wherein: -X, Y, Y ', W and W'have the same meanings as defined above; -Z is O (where Z is a 1,3-relationship of the central chain); Represents a bond between 3 carbon atoms), a saturated or unsaturated hydrocarbon chain of 1, 2, 3 or 4 atoms, all of which are heteroatoms such as oxygen, sulfur and nitrogen. be replaced by atoms and / or may be _{replaced, -R 1, R 2, R} 2 ', R 3, R 3', is _{R 4, R 4 ', R} 5, R 5' · them, Which may be the same or different and represents hydrogen or lower alkyl such as methyl, ethyl or n-propyl].

Of these, the type [Wherein: -n is an integer equal to 2 or 3; -X is the following vitamin D side chain moiety: (4-hydroxy-4-methyl) pentyl, (R)-or (S)-( Three
-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; 4,5-epoxy, 4-methyl-2-
Pentynyl; 4-hydroxy-4-ethyl-2-hexynyl; (3-methyl-2,3-epoxy) -butyloxy;
(3-hydroxy-3-ethyl) pentyloxy; (4
--Hydroxy-4-ethyl) -hexyloxy, --Y, Y ', W and W'are the same and represent hydrogen, or taken together 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 ′ can be the same or different and are hydrogen or methyl. A cyclic derivative of

─────────────────────────────────────────────────── ─── Continuation of the front page (72) Inventor Schnoes, Heinrich Kay 53705, Madison, Summit Avenue, Wisconsin, USA 1806 (72) Inventor Kai, Zee Yang Wisconsin 53703, Madison, East Goham 711 (72) 72) Inventor Phelps, Merry E 53589, Wisconsin, USA, Stoughton, West Prospect Avenue 116 (72) Inventor Smith, Connie M, USA 53713, Wisconsin, Madison, Post Road 2918 (56) References Akira 55-500428 (JP, a) PCT National Akira 54-500080 (JP, a) (58 ) investigated the field (Int.Cl. ^7, DB name) CA (STN) REGISTRY ( TN)

Claims (4)

(57) [Claims] 1. A 1α, 25-dihydroxyvitamin D ₃ -1,3,2
5-trihexanoate. 2. A 1α, 25-dihydroxyvitamin D ₃ -1,3,2
5-trinonanoate. 3. The formula: A vitamin D compound having {wherein X ¹ and X ² independently represent hydrogen or --CH ₃ CO} and has a hydroxy group at carbon 25 and has a desired therapeutic activity in vivo. Selecting the vitamin D compound; when the carbon 25 is attached to the oxygen of the hydroxy group, it is hydrolyzed to the active vitamin D compound at a desired rate in vivo to produce the active vitamin D compound. Modified vitamin D that will prolong the activity profile in the body
Selecting a -CH ₃ CO group would provide derivatives;
And modifying the vitamin D compound by derivatizing a hydroxy group at carbon 25 with the —CH ₃ CO group, the formula: {Wherein X ¹ and X ² are as described above, and X ³ is −
A method of controlling the activity profile of a vitamin D compound comprising obtaining a modified vitamin D derivative having CH ₃ CO. 4. The formula: A vitamin D compound having {wherein X ¹ and X ² independently represent hydrogen or --CH ₃ CO}, having a --OH group attached at carbon 25, and having a therapeutic activity profile in vivo. And (a) a -OX ³ group (where X ³ is -CH ₃ CO, which can be hydrolyzed to a -OH group at carbon 25 at a desired rate in vivo. And (b) a —OX ³ group attached at carbon 25, where X ³ is —CH ₃ CO 2.
Providing a modified form of the Vitamin D compound according to claim 1, wherein the activity profile of the Vitamin D compound is lengthened by prolonging the activity profile of the Vitamin D compound.