JPS6075481A - Lactone derivative-cyclodextrin clathrate - Google Patents

Abstract

NEW MATERIAL:Cyclodextrin clathrate of a compound of formula I (A is formula II and formula III). EXAMPLE:2, 3-Seco-2a-oxa-14-hydroxy-14beta-card-20(22)-enolide-3-one-gamma-cyclod extrinclathrate. USE:Cardiotonic: it has positively inotropic action of increasing contraction force of heat muscles. It has high solubility in water, long duration of pharmacological actions almost without influence on pulse rate. PREPARATION:A compound of formula IV is dissolved in a solvent such as chloroform or benzene, then a peracid such as perbenzoic acid or m-chloroperbenzoic acid is added to effect oxidation at room temperature to reflux one of the solvent, preferably 60-70 deg.C to give the compound of formula I .

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to novel cyclodextrin clathrate compounds. More specifically, the present invention provides a lactone derivative characterized by including a lactone derivative represented by the general formula -
Pertains to cyclodextrin clathrate compounds.

The lactone derivative represented by the above general formula (I) is a new compound and has the effect of increasing myocardial contractility (positive inotropic effect), and can be used as a cardiotonic agent for the treatment of heart diseases such as congestive heart failure. It is a useful compound. The clathrate compound of the present invention has excellent water solubility and the above-mentioned pharmacological action has a long duration. Furthermore, the clathrate compound of the present invention has almost no effect on heart rate, and conventional cardiac inotropes such as digitalis have a narrow safety margin, and may cause digitalis toxicity, premature contractions, atrioventricular block, etc. Although electrocardiogram abnormalities and side effects on the gastrointestinal system and nervous system were observed, the clathrate compound of the present invention is characterized in that the above-mentioned side effects are extremely weak compared to conventional cardiotonic agents such as digitalis.

In the present invention, the lactone derivative used is a new compound and can be produced by various methods, but one preferred example thereof is the method shown in Reaction Scheme-1 below.

Reaction Scheme-1 t) (If) In formula C, R1, R2 and R3 are the same as above. ] The reaction shown in the above reaction scheme-1 is a novel reaction discovered by the present inventors, and the compound (I) of the present invention has the general formula (II
) is produced by reacting the compound represented by the formula with peracid. That is, the compound of general formula (II) may be dissolved in a suitable inert solvent, and then an organic peracid may be added to this solution to carry out an oxidation reaction with the organic peracid. As the organic solvent, a wide range of organic solvents can be used without particular limitation as long as it can dissolve the compound to be treated and the organic peracid. Specific examples thereof include halogenated hydrocarbons such as chloroform, methylene dichloride, carbon tetrachloride, and trichloroethylene, aromatic hydrocarbons such as benzene and toluene, and halogenated hydrocarbons. In addition, it can be widely used as an organic peracid without particular limitation, such as perbenzoic acid, metachloroperbenzoic acid, metanitroperbenzoic acid,
Examples include perbenzoic acids such as varanitroperbenzoic acid and 3,5-dinitroperbenzoic acid.

The amount of organic peracid used is usually 1 to 3 times, preferably 1.2 to 1.5 times, the amount of compound (II). In the present invention, the organic peracid may be added as it is, or may be added dissolved in a solvent in advance. As the solvent used here, the same organic solvents as mentioned above can be mentioned. The above oxidation reaction is usually carried out at room temperature to the reflux temperature of the solvent, preferably 60 to 70°C.
The oxidation reaction progresses suitably and is generally completed in 20 minutes to 24 hours. The target compound thus obtained can be easily isolated and purified from the reaction mixture by conventional isolation methods such as recrystallization, solvent extraction, column chromatography such as high performance liquid chromatography, and distillation.

Incidentally, the lactone derivative naturally includes optical isomers.

The cyclodextrin used as the host molecule in the present invention is not particularly limited, and α-cyclodextrin, β-cyclodextrin, or γ-cyclodextrin may be used, or mixed dextrin may be used. Particularly in the present invention (5), it is preferable to use β-cyclodextrin or γ-cyclodextrin.

The lactone derivative-cyclodextrin clathrate compound of the present invention is produced, for example, by the method shown below.

First, cyclodextrin is dissolved in water or suspended in the form of a slurry to prepare 1 ill of an aqueous solution or suspension of cyphlobuxtrin. Next, the above-mentioned lactone derivative may be added to the aqueous solution or suspension as it is, or a solution obtained by dissolving the derivative in an organic solvent compatible with water may be added, and the mixture may be thoroughly shaken or kneaded.

Examples of organic solvents that are compatible with water include alcohols such as methanol, ethanol, and isopropanol, aprotic solvents such as dimethylformamide and dimethylacetamide, acetone, acetonitrile, and diethyl ether.

The blending ratio of the lactone derivative represented by the general formula (I) and cyclodextrin is not particularly limited and can be appropriately selected within a wide range, but it is usually preferable to use an equimolar amount or more of the latter to the former. Shaking or mixing is usually done at room temperature
It is best to carry out the reaction at around 150°C, preferably around 6-100'C. The above-mentioned shaking or mixing is usually carried out for about 30 minutes to 2 hours, and then the reaction mixture is filtered or concentrated under reduced pressure to produce the desired clathrate compound of the present invention.

The clathrate compound of the present invention has improved solubility in water and also has a longer duration of pharmacological action.

The clathrate compounds of the present invention can be administered to animals and humans as such or together with conventional pharmaceutical carriers. The dosage unit form is not particularly limited and can be appropriately selected and used as required. Such dosage unit forms include tablets, granules,
Examples include oral preparations such as oral solutions and parenteral preparations such as injections. The active ingredient to be administered is not particularly limited and can be appropriately selected from a wide range, but in order to achieve the desired effect, it should be administered in an amount of 0.001 to 1 liter per kilogram of body weight per day.
It is better to set it to l1g. It is also preferable that the dosage unit form contains 0.01 to 25 ma of the active ingredient.

In the present invention, oral preparations such as tablets, capsules, and oral solutions are manufactured according to conventional methods. For example, tablets are formed by mixing the compound of the invention with conventional pharmaceutical excipients such as gelatin, starch, lactose, magnesium stearate, talc, gum acacia, and the like.

Capsules are prepared by mixing the compound of the present invention with an inert pharmaceutical filler or diluent, and filling the mixture into hard gelatin capsules, soft capsules, and the like. A syrup or elixir is prepared by mixing the compound of the present invention with a sweetening agent such as sucrose, a preservative such as methyl- and propylparabens, a coloring agent, a seasoning agent, and the like. Moreover, parenteral preparations are manufactured according to conventional methods. For example, when manufactured as an injection, it is preferable that solutions and suspensions be sterilized and isotonic with blood; when forming these solutions, emulsions, and suspensions, dilution All agents commonly used in this field can be used, such as water, ethyl alcohol, propylene glycol, ethoxylated isostearyl alcohol, polyoxylated isostearyl alcohol, polyoxyethylene sorbitan fatty acid esters, and the like.

In this case, a sufficient amount of salt, glucose, or glycerin may be included in the pharmaceutical preparation to prepare an isotonic solution, and usual solubilizing agents, buffers, soothing agents, etc. may be added. It's okay. Furthermore, coloring agents, preservatives, perfumes, flavoring agents, sweeteners, etc., and other pharmaceuticals may be included in the pharmaceutical preparation, if necessary. To further increase stability, parenteral formulations can be frozen after filling and water removed by freeze drying techniques known in the art. The lyophilized powder can then be reconstituted immediately before use.

Reference examples, examples, and pharmacological test results are shown below.

Reference example 1 Digitoxigenin lactone synthesis Digitoxigenin 1.10 (+ anhydrous chloroform 15
Add MCPBAl, 00 (l) to the Peng solution and add 2 liters of MCPBAl under reflux.
Stirred for 4 hours. The reaction solution was washed successively with a 5% aqueous sodium hydrogen carbonate solution and water, dried over magnesium sulfate, and then the solvent was distilled off to obtain a residue 1.24Q. This product was subjected to chromatography on 100 g of silica gel using an ethyl acetate-chloroform solvent system as a developing solvent while increasing the ethyl acetate concentration sequentially. A mixture of lactones 946 with a melting point of 225-235°C was obtained from the 70% ethyl acetate-chloroform eluate.
11(+(83,0%)) was obtained. This mixture was 2,3-
Seco-2a-oxa-14-hydroxy-14β-card-20(22)-ephride-3-one (hereinafter referred to as compound (Ia) J) and 3,4-seco-38-oxa-
14-hydroxy-14β-card-20(22)-ephride-3-one (hereinafter referred to as "Compound (Ib) J")
and in a ratio of about 1:1, respectively.

Compound (Ia) Compound (Ib) Mixed melting point of 10-digitoxigenin lactone: 225-235°C IR (KBr, cr'): 3500.2975.2900, 1790°174.5
．． 1735.1625.1455゜1310.1295
,1190,1165゜1130.1070.1035 LJV(λll1ax, MeOH): nm(l
og ε)=218.0(4,16>obtained(Ia
) and (H) )-IP
Lc (LS-410-ODS 2..1cmX 30
cm, acetonitrile:methanol:water=10nia0:
85, flow rate 9.9mQ/min, detection light 240nll
) to separate. The retention time is 62.5 minutes for (1a), (I
b) The peak at 69 minutes was fractionated and 15+no was obtained for each.

Compound (Ia) Melting point: 235-236°C tJV (λn+ax, MeOH): 218r
+m (C15000) I R (KBr, cr'): 3525.3150.2900.1790°1742.
1720.1630.1472゜1450.1390.
1370.1290°1255.1210.1190,
1160°1115.1080.1068.1030°1000.980,950,890,850°834.
805.740cr' MS (E I) : m /z (rel, int
, ) 388 (M+, 39), 370 (5).

352 (2), 278 (9), 260 (19).

235 (75), 217 (86).

194 (32), 175 (38).

121 (59), 95 (68), 81 (74).

79 (71), 69 (56), 67 (77).

55 (96), 41 (100) Fast dissolution MS: Actual value: 388.2257 Calculated value: 388.2250 (as C23Hs 205) [α]p 47° (c 0.35.CHCQ3)H
'NMR (400MHz, CDCQa) 0.90
(3H,s), 1.01 (3H,s).

1.37 (IH, OH).

1.57 (IH, ddd, J-16, 5°10.4
．． 1. 2H2).

1.82 (1H, Ill).

2.04 (11-1, dd, J=16.5.6.1
H2).

2.24 (IH, d, J-13, 1Hz).

2.80 (IH, m).

3.1 2 (IH, dd, J-13, 1, 13, 1H
2).

4.07 (11-1, ddd, J-13, 1, 6, 1
゜1.2Hz).

4, 27 (IH, dd, J=13. 1. 10.
4H2).

4.82 (1)-1,dd, J-18,0,1,5H
z).

13- 4.91 (IH, dd, J=18.0.1.5H2)
.

5, 89 (1H,t, J-1,5H7)OF) [
+1 Compound (i) Melting point = 264-266°C UV (λwax, MeOH): 218ni(
ε13600) IR (KBr, c+a-'): 3550.31
40.29B0,2910°1790.1750,17
35.1625°1475.1404.1390.13
70°1355.1345.1312,1275°12
60.1223, 1190.1170゜1130.10
70,1050,1030゜994.955,905,
890. 865°83001' MS (E I): m /z (rel, int
, )38B (M+, 57), 370 (3), 3
52(1), 291(13), 278(9).

235 (48), 217 (17).

14-207 (26), 194 (100), 175 (33
), 134. (23), 95 (34).

83 (11), 79 (41), 69 (36).

55 (4,7), 41 (58) High speed dissolution MS; Actual value: 388.2253 Subtraction value: 388.2250 (C2s Hs Q O5
) (α]p 51.6” <00.63.CHCQ3 )
H' NMR (400MH7, CDC03) 0.90
(3H,s >, i, 03 (3H,s ).

1.19 (1H, OH).

1.46 (IH, dd, J=15.2.13.4Hz
).

1.81 (IH, m >, 1.96 (IH, dd.

J-15,2,7,6)1z).

2.41 (1H, dd, J=15.2.7.6Hz>
.

2.73 (IH, dd, J=15.2.13.4Hz
> 2.80 (IH, m) 3.95 (IH, dd, J-13,1,1,5l-1
z) 4.58 (IH, dd, J=10.0.13.1)
-1z) 4.81 <IH, dd, J-18,0,1,5Hz) 4, 97 (IH, dd,,) - 18. 0.
1. 5l-(z) 5, 90 (IH,t, J-1,5H7)Dl)I
II Example 1 5011 g of Compound (Ia) obtained in Participant Example 1 and 400 I1 g of γ-cyclodextrin were heated and dissolved in 1011 G of water, allowed to stand at room temperature overnight, and the precipitated crystals were collected. )-γ-cyclodextrin inclusion compound 3
80111 was obtained.

mp, 289-290℃ Colorless fine prism-like product.

This clathrate includes compound (Ia) and compound (Ia
)/γ-cyclodexteryl 1/2 (molar ratio).

Compound (Ia) and γ-cyclodextrin are 1=2(
molar ratio) means that compound (Ia) appears at 65.95 in its NMR spectrum in heavy water at 22
This was determined based on the fact that the integral ratio of γ-cyclodextrin (8 molecules of dextrin are condensed to form a ring) appearing at proton 65.05 with the 01 proton 17- is 1:16.

NMR (D20): 60.87 (3H,s), 1.
00 (3H, s), 1, 1~2.3 (br, m), 2.87 (IH, dd, J+ = 8.6H2, J2-
4.8Hz), 3.13 (IH,br,t,J=13.1Hz>,
3.54 (16H,t, J-9,5H2), 3.6
1 (16H, dd, J+ = 9.9Hz.

J2-3.7H2), 3,71 (16H,br, dd, J+ =9.9
Hz.

J2-1.81 (z), 3.8 (481-1, Il
).

4.14 (IH, dd, J+ = 13.1Hz.

18- J2 ~5. 1H2), 4. 24 (1H, dd.

J+ ~13. 1Hz, J2 ~10. 5Hz
), 5.05 (16)-1,d, J=3. 7H
z), 5.95 (1H,s) IR(KBr, am-1): 3370, 2950. 1730. 1625°11
60, 1020. 940 Example 2 From Compound (Ib) 50+11(+ obtained in Reference Example 1) and γ-cyclodextrin 4001B, 360 mg of Compound (Ill>-γ-cyclodextrin clathrate compound) was obtained in the same manner as in Example 1.

mp, 275-276℃ Colorless prismatic crystals.

This clathrate includes compound (Ib), compound (Ib), and compound (Ib).
b)/γ-cyclodexteryx=1/2 (molar ratio). As in Example 1, it was determined from the integral ratio of protons at δ5.05 and δ5.95 in the NMR spectrum.

NMR (D20): δ0.88 (3H,s), 1.
00 (3H,S), 1, 2~1. 9 (br, m), 2, 08
(n+), 2. 28 (+n), 2, 70 (I
H, t, J=13. 8 t1z), 2.88 (
1H, dd, J+ ~9. 5)-1z.

J2 ~3. 7H2), 3, 54. (16H,t, J-9,5H2)3.6
2 (16H,d(1,J+ ~9.9Hz.

J2 ~3. 7H2), 3.70 (16H, br, dd, J+-9,9Hz
.

J2 ~ 1.8H7), 3.8 (48H, m), 3.97 (IH, dd.

J+ ~13.1Hz, J2 -1. 2)-1z
), 5.05 (16H, d, J-3,7Hz)
, 5.95 (IH,s) IR(KBr, am-1): 3370, 2950. 1730. 1625°11
60, 1025.940 Example 3 Compound (Ib)-β-cyclodextrin clathrate compound 21O was prepared in the same manner as in Example 1 from compound 50111g and β-cyclodextrin 321 rQO.
n+g was obtained.

mFl, 269-270°C Colorless prismatic crystals.

This clathrate includes compound (Ib), compound (Ib), and compound (Ib).
b)/ββ-cyclodextrin=1/2 molar ratio). As in Example 1, it was determined from the integral ratio of protons at δ5.03 and δ5.88 in the NMR spectrum.

NMR (D20): δ0.89 (3H,s), 1.
13 (38,s), 1,25-2.4 (br, m), 2.76 (I
H, br, t, J=13Hz), 2.93 (IH
, dd, J+ ~9.5Hz, J2=3.7H2), 3.55 (14H,t, J=9.5Hz), 3.
63 (14H, dd, J+ ~9.8H7.

J2~3.6t-1z), 3.71 (14H, br, d, J=9.7Hz)
, 21-3.8 (42H, m), 4, 14 (1)-1, br, d, J-13l-1z
), 5.03 (14H,d, J=3.6Hz
) 5.88 (IH, s) I R (KBr, am”): 3370, 2950. 1740. 1630°11
55, 1025.940 Example 4 5011 g of compound (Ia) and β-cyclodextrin 3
From 21 mg, the compound (Ia
)-β-cyclodextrin clathrate compound (2901 g) was obtained.

B, 273-274℃ Colorless prismatic crystals.

This clathrate compound includes compound (Ia) and compound (Ia
)/β-cyclodextrin-1/2 (molar ratio). As in Example 1, it was determined from the integral ratio of protons at δ5.02 and δ5.88 in the NMR spectrum.

NMR (D20): δ0.88 (3H,s), 22
- 1, 11 (3H,s), 1. 3 ~2. 4 (br, m), 2.91
(IH, dd, J+ = 8.6Hz, J2-4.8H
2), 3.15 <11-1. br, t, J=13.1l-
(Z), 3.55 (14, H, t, J= 9.4
Hz), 3.62 (14H, dd, J+ = 9.8
Hz.

J2 = 3.6) - 1z), 3.73 (14H, br, d, J = 9.8H2),
3, 8 (4-28,m), 5.02 (14th, d, J-3,6)1z)5.
88 (IH, s) IR (KBr, cm-'): 3370.2960, 1730.1630°1160.
1020.940 Example 5 <Calibration curve> Cyclodextrin clathrate compounds (compounds of Examples 1, 2, 3, and 4) were weighed and made into an aqueous solution. Compound (Ia
> and (Ib) were taken as acetonitrile-water (1:1) solutions and were taken as the lactone concentration in the solution. 0.1-2111
0/rm was set. Analysis was performed using high performance chromatography (1-1ypersil OD Sφ4.6X25O
1I1m column, eluent niacetonyl-lilu water (1:
1), flow rate 1, 5mQ/min, detector UV24O
r+m) to create a calibration curve (Figure 11 and 2).
figure).

<Solubility measurement> γ-Cyclodextrin clathrate compounds (compounds of Examples 1 and 2) and γ-cyclodextrin clathrate compounds (compounds of Examples 3 and 4) were each given 50+no, and water was added once to this. Ta. Compounds (Ia) and (Ib) were taken at 10 ma, and 1 h of water was added thereto. Place in a constant temperature bath, incubate for 20 minutes, centrifuge, and remove the supernatant from 1 to 5μ
Analyzed using Q-filter and high-speed liquid chromatography.

The amount of compound (Ia) or (Ib) dissolved was calculated from the calibration curve. The results are shown in FIGS. 3 and 4.

As is clear from FIGS. 3 and 4, it can be seen that the clathrate compounds of the present application have an 8- to 10-fold increase in solubility in water compared to the lactone derivatives (Ia and Ib) alone.

<Pharmacological test> Male and female hybrid adults weighing 9 to 15 ka are anesthetized by intravenously administering bendoparbital sodium salt 30n+a/ko. Subsequently, Ben I parbital sodium salt 41Ω/ka/hr was continuously injected intravenously to maintain a constant depth of anesthesia. with ventilator - 18 minutes
Artificial respiration was performed at a respiratory rate of 20 mG/ka and an inspiratory force of 20 mG/ka, and the chest was opened.

Left ventricular contractile force was measured via an arched strain gauge attached to the outer wall of the left ventricle. A polyethylene tube was inserted into the left femoral artery, systemic blood pressure was measured via a pressure transducer, and a heart rate tachometer was driven by the blood pressure pulse wave to measure heart rate. Electrocardiograms were recorded in standard lead II.

All parameters were recorded on an ink recorder.

The test compound was continuously infused intravenously using an infusion pump. The inotropic effect of the test compound was expressed as a % change in the tension generated before administration of the 25-compound.

Table 1 shows the results.

Table 1 Figures 1 and 2 show compounds (Ia) and (Ib), respectively.
It is a graph showing the relationship between the concentration of > and the peak height of LJV2 to. Figures 3 and 4 show the compounds (
Solubility curves of Ia) and (Ib).

FIG. 5, FIG. 6, FIG. 7, and FIG. 8 are NMR spectra of the clathrate compounds obtained in Example 1.2.3.4, respectively. Figures 9 and 10 show the IR of the clathrate compound obtained in Example 3.4, respectively.
It is a spectrum diagram.

(That's all) 27- Figure 1 Ihi, 6'F') (IQ) Uno! (mg/tenl) 2nd
Figure 4'rL'# substance (Ib) 04 degrees (mg/ml) -
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Claims (1)

[Claims] ■ Show. ) A lactone derivative-cyclodextrin clathrate compound characterized by including a lactone derivative represented by: